Archive for the ‘ regeneration ’ Category

Older adults who favored this eating style lost less brain volume, study finds

Source: More Signs Mediterranean Diet May Boost Your Brain – News – Health.com

Ten years ago, Shinya Yamanaka revolutionized biological research with his discovery of how to turn ordinary skin cells into stem cells with just four key genes.

Source: Reflecting on the Discovery of the Decade: Induced Pluripotent Stem Cells | Gladstone Institutes

Mice were rejuvenated through a four-gene cocktail that allowed them to repair aging signs including loss of hair and organs malfunction. There were no signs of cancer and compared to untreated mice, the reprogrammed mice looked younger, with better organ function, improved cardiovascular performance and lived 30 percent longer.

Source: Study shows aging might be reversible thanks to cells group

February 13, 2013

Shocking Alien Fears Force Pope From Office

By: Sorcha Faal, and as reported to her Western Subscribers

 

A stunning Ministry of Foreign Affairs (MFA) report prepared for President Putin, which is circulating in the Kremlin today, states that Pope Benedict XVI was forced to resign this past week over Catholic Church fears that this 85-year-old leader of over 1 billion Christians was “mentally and physically unprepared” to deal with the coming revelation about the truth of alien beings.

In our 22 January report, Russia Orders Obama: Tell World About Aliens, Or We Will, we detailed how the issue of extraterrestrial beings was brought to the forefront of the World Economic Forum (WEF) with the naming in their 2013 Executive Summary of the danger posed to our world over the discovery of alien life with their stating: “Proof of life elsewhere in the universe could have profound psychological implications for human belief systems.”

Also noted in our previous report were Prime Minister Medvedev’s 7 December 2012 off-air comments to reporters which were recorded and wherein he stated: “Along with the briefcase with nuclear codes, the president of the country is given a special ‘top secret’ folder. This folder in its entirety contains information about aliens who visited our planet… Along with this, you are given a report of the absolutely secret special service that exercises control over aliens on the territory of our country… More detailed information on this topic you can get from a well-known movie called Men In Black… I will not tell you how many of them are among us because it may cause panic.”

Spurring Pope Benedict XVI to become the first leader of the Catholic Church to resign in nearly 600 years, this MFA report says, was the appearance over Los Cristianos, Spain on 21 August 2011 of the long prophesized “bird of prey” interplanetary spacecraft, and which was followed nearly 3 weeks ago with a fleet of them appearing in the skies over Mexico City.

To fully understand the significance of these “bird of prey” UFO’s, this report continues, files relating to the 27 September 1989 Voronezh Incident must be studied in length, especially as it relates to the “messages” delivered to eyewitnesses from the “giants”.

In an 11 October 1989 New York Times article about the Voronezh Incident titled U.F.O. Landing Is Fact, Not Fantasy, the Russians Insist it says:

“It is not a joke, nor a hoax, nor a sign of mental instability, nor an attempt to drum up local tourism by drawing the curious, the Soviet press agency Tass insisted today in discussions of what it called an extraterrestrial visit to southern Russia.

Residents of the city of Voronezh insisted today that lanky, three-eyed extraterrestrial creatures had indeed landed in a local park and gone for a stroll and that a seemingly fantastic report about the event carried Monday by the official press agency Tass was absolutely true.

The three-eyed creature, about nine feet tall and fashionably dressed in silvery overalls and bronze boots and with a disk on its chest, disappeared, then landed and came out for a promenade with a companion and a robot.

The aliens seemed to communicate with each other, producing the mysterious appearance of a shining triangle, and activated the robot with a touch.”

Regarding these “messages” from the Voronezh “giants”, this MFA report says, was the warning to human beings that when these “bird of prey” UFO’s descend upon Earth the whole planet will be in peril.

The Voronezh “giants” further related, this report says, that the alien beings associated with these “bird of prey” UFO’s were the cause of the 14 April 1561 massive “sky battle” over Nuremberg, Germany which was depicted in a famous 16th century woodcut by Hans Glaser [photo 3rd left] and described by the residents as: “A very frightful spectacle.” “The sky appeared to fill with cylindrical objects from which red, black, orange and blue white disks and globes emerged. Crosses and tubes resembling cannon barrels also appeared whereupon the objects promptly began to fight one another.”

Important to note is that the Catholic Christian faith headed by Pope Benedict XVI, as well as nearly every other religion on Earth, all prophesize in their teachings a time when the “gods” will return to our planet and engage in a battle that could very well bring our entire planet to the brink of destruction.

Equally important to note about Pope Benedict XVI’s shock resignation is how it eerily compares with Saint Malachy, who as an Irish saint and Archbishop of Armagh, in the 12th Century, received a vision of 112 Popes later attributed to the apocalyptic list of Prophecy of the Popes. He was the first Irish saint to be canonized by Pope Clement III in 1199.

American authors Tom Horn and Cris Putnam in their 2012 book “Petrus Romanus: The Final Pope is Here” about Saint Malachy’s prophecies told interviewers last Spring that Pope Benedict XVI would resign by late 2012, or early 2013, and described the next Pope to follow as “Petrus Romanus,” or “Peter the Roman,” writing: “In the final persecution of the Holy Roman Church there will reign Peter the Roman, who will feed his flock among many tribulations; after which the seven-hilled city will be destroyed and the dreadful Judge will judge the people.”

Though the masses of people reading of the things this report contains will, undoubtedly, ridicule them, the same cannot be said of the elite moneyed classes who, even at this writing, are protecting themselves from “something” at such a fever-pitched pace it is destabilizing the entire global economy, and as exampled by the highly respected Zero Hedge news service in their article titled “What Do They Know That We Don’t?” and which, in part, says:

“Friday evening when no one was supposed to pay attention, Google announced that Executive Chairman Eric Schmidt would sell 3.2 million of his Google shares in 2013, 42% of the 7.6 million shares he owned at the end of last year—after having already sold 1.8 million shares in 2012. But why would he sell 5 million shares, about 53% of his holdings, with Google stock trading near its all-time high?

“Part of his long-term strategy for individual asset diversification and liquidity,” Google mollified us, according to the Wall Street Journal. Soothing words. Nothing but “a routine diversification of assets.”

Routine? He didn’t sell any in 2008 as the market was crashing. He didn’t sell at the bottom in early 2009. And he didn’t sell during the rest of 2009 as Google shares were soaring, nor in 2010, as they continued to soar. In 2011, he eased out of about 300,000 shares, a mere rounding error in his holdings. But in 2012, he opened the valves, and in 2013, he’d open the floodgates. So it’s not “routine.”

Mr. Schmidt isn’t alone. Corporate insiders were “aggressively selling their shares,” reported Mark Hulbert. And they were doing so “at an alarming pace.” The buy sell-to-buy ratio had risen to 9.2-to-1; insiders had sold over 9 times as many shares as they’d bought. They’d been aggressive sellers for weeks.

Instantly, soothing voices were heard: “don’t be alarmed,” they said. But Mr. Schmidt and his colleagues at the top of corporate America, multi-billionaires many of them, are immensely well connected, not only to each other but also to the Fed, whose twelve regional Federal Reserve Banks they own and control.”

To why Google Chairman Schmidt did not attend this years World Economic Forum, where the danger of aliens was being discussed, opting instead for a visit to North Korea (who announced yesterday that they had exploded another nuclear weapon) and when coupled with the information contained in this MFA report, is far from being “soothing”, and is, instead, something well all should be very alarmed about as the end is much nearer than the beginning as those with “eyes to see” and “ears to hear” already know.

February 13, 2012 © EU and US all rights reserved. Permission to use this report in its entirety is granted under the condition it is linked back to its original source at WhatDoesItMean.Com. Freebase content licensed under CC-BY and GFDL.

[Ed. Note: Western governments and their intelligence services actively campaign against the information found in these reports so as not to alarm their citizens about the many catastrophic Earth changes and events to come, a stance that the Sisters of Sorcha Faal strongly disagrees with in believing that it is every human beings right to know the truth. Due to our missions conflicts with that of those governments, the responses of their ‘agents’ against us has been a longstanding misinformation/misdirection campaign designed to discredit and which is addressed in the report “Who Is Sorcha Faal?”.]

You May Already Be To Late…But It Has Begun!

They Are Going To Come For You…Why Are You Helping Them?

Source: Shocking Alien Fears Force Pope From Office

Mechanisms and methods of methonine restriction

Life Extension Benefits of Methionine Restriction

by Ben Best

CONTENTS: LINKS TO SECTIONS BY TOPIC

  1. METHIONINE BASICS
  2. METHIONINE RESTRICTION EFFECTS
  3. METHIONINE RESTRICTION FOOD DATA
  4. METHIONINE RESTRICTION DIET
HEART MUSCLE METHIONINE
[HEART MUSCLE METHIONINE]

I. METHIONINE BASICS

Methionine is the only essential amino acid containing sulfur. Methionine is the precursor of the other sulfur-containing amino acids: cysteine, taurine, homocysteine, and cystathione. Methionine is essential for the synthesis of proteins and many other biomoleules required for survival. Rats fed a diet without methionine develop fatty liver disease which can be corrected by methionine supplements [DIGESTIVE DISEASES AND SCIENCES; Oz,HS; 53(3):767-776 (2008)]. Dietary methionine is essential for DNA methylation. Reduced DNA methylation results in genetic instability, aberrant gene expression, and increased cancer.

The above paragraph is the first paragraph from the section on methionine in my article dealing with the Methionine Cycle. Material in that article is useful background for the information below. Note, however, that there is an inverse correlation between lifespan and the methionine content of protein in the heart muscle of eight mammalian species [MECHANISMS OF AGEING AND DEVELOPMENT; Ruiz,MC; 126(10):1106-1114 (2005)]. The sulfur-containing amino acids methionine and cysteine are the most readily oxidized of any of the amino acids — both as free amino acids or in proteins. Methionine is oxidized to methionine sulfoxide, but methionine sulfoxide reductases enzymatically regenerate methionine [BIOPHYSICA ET BIOCHEMICA ACTA; Lee,BC; 1790 (11): 1471-1477 (2009)].

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II. METHIONINE RESTRICTION EFFECTS

Substantial evidence indicates that as much as half of the life-extension benefits of CRAN (Calorie Restriction with Adequate Nutrition) are due to restriction of the single amino acid methionine. In a study of rats given 20% the dietary methionine of control rats, mean lifespan increased 42% and maximum lifespan increased 44% [THE FASEB JOURNAL;Richie,JP; 8(15):1302-1307 (1994)]. Blood glutathione levels were 81% higher in the methionine-restricted rats at maturity, and 164% higher in old age. In other studies, methionine-restricted rats showed greater insulin sensitivity and reduced fat deposition [AMERICAN JOURNAL OF PHYSIOLOGY; Hasek,BE; 299:R728-R739 (2010) and AGING CELL; Malloy,VL; 5(4):305-314 (2006)].

An experiment on mice given 35% the methionine of controls showed only a 7% increase in median life span [JOURNALS OF GERONTOLOGY; Sun,L; 64(7):711-722 (2009)]. Another mouse study showed lowered serum insulin, IGF−1, glucose, and thyroid hormone for methionine at one-third the normal intake. There was significant mouse mortality for methionine less than one-third normal intake, but with one-third intake of methionine maximum lifespan was significantly increased [AGING CELL; Miller,RA; 4(3):119-125 (2005)]. Rats generally show greater longevity benefits from CRAN than mice.

Mitochondrial free radical generation is believed by many biogerontologists to be a significant contributor to aging damage. Rats given 20% the dietary methionine of control rats show significantly decreased free radical generation from complex I and complex III of liver mitochondria as well as from complex I of heart mitochondria — associated with reduced oxidative damage to mitochondrial DNA and protein [THE FASEB JOURNAL;Sanz,A; 20(8):1064-1073 (2006)]. These results are comparable to the reduced mitochondrial free radical generation seen in CRAN rats [ENDOCRINOLOGY; Gredilla,R; 146(9):3713-3717 (2005)]. Rats given 60% rather than 20% of the methionine of control rats showed nearly the same amount of reduced mitochondrial free radical generation and damage [BIOCHEMICA ET BIOPHYSICA ACTA; Lopez-Torres,M; 1780(11):1337-1347 (2008)]. Body weight was not reduced with 60% dietary methionine, leading to the conclusion that such reduction would not result in reduced growth in children [REJUVENATION RESEARCH; Caro,P; 12(6):421-434 (2009)]. It was concluded that methionine restriction is the sole reason for reduced mitochondrial free radical generation and damage associated with CRAN [Ibid.] and protein restriction [BIOGERONTOLOGY; Caro,P; 9(3):183-196 (2008)].

Evidence for the suggestion that methionine oxidation plays a significant role in lifespan can be found in the considerable lifespan extension benefits seen in transgenic fruit flies that overexpress a gene for repairing oxidized methionine in protein [PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES (USA); Ruan,H; 99(5):2748 (2002)]. The sulfur-containing amino acids methionine and cyteine are more easily oxidized in proteins than other amino acids [JOURNAL OF PHYSIOLOGY)], which is apparently related to the reduced free radical generation in mitochondria seen in methionine restriction. Both the fruit fly experiment and the methionine restriction experiments indicate a significant impact on lifespan from methionine oxidation.

It has been suggested that glycine supplementation has the same effect as methionine restriction. An experiment with glycine supplementation in rats showed a 30% extension in maximum lifespan [FASEB JOURNAL; Brind,J; 25:528.2 (2011)]. Additionally, three grams of glycine daily has been shown to improve sleep quality in young (average age 31) female Japanese adults [SLEEP AND BIOLOGICAL RHYTHM; Inagawa,K; 4:75-77 (2006)]

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TABLE 1 Lysine, Threonine and Methionine in Food
(milligrams amino acid per gram protein)
Food Sulfur-containing amino acid Lysine Threonine
Nuts,Seeds 46 ± 17 45 ± 14 36 ± 3
Animal foods 38 85 ± 12 44
Cereals 37 ± 5 31 ± 10 32 ± 4
Fruits 27 ± 6 25 ± 12 29 ± 7
Legumes 25 ± 3 64 ± 10 38 ± 3

III. METHIONINE RESTRICTION FOOD DATA

The adjoining table (my Table 1) from [AMERICAN JOURNAL OF CLINICAL NUTRITION; Young,VR; 59(suppl):1203s-1212s (1994)] indicates the essential amino acids most likely to be limited in plant protein foods. Cereal protein contains comparable sulfur-containing amino acids (including methionine) per gram as animal foods, whereas fruit and legume protein contain about 65% as much methionine. Nuts and seeds are particularly high in methionine, on average 20% higher in methionine than animal protein, although the absolute amount of protein in animal foods tends to be higher, which makes total methionine intake generally higher in animal foods. Vegetables are not shown in Table 1, but as described in Table 4 in the AMERICAN JOURNAL OF CLINICAL NUTRITION paper from which Table 1 is taken, vegetables are on average in the 1-2% range for percent protein and fruits are in the 0.5-1% protein range — so neither fruits nor vegetables should be considered serious sources of protein (green peas are an exceptional vegetable with 5.4% protein, and avacado is an exceptional fruit with 2% protein). Cereals are typically 7-13% protein and legumes are typically 20-30% protein (soybeans are exceptionally high in protein even for legumes, being in the range of 35-45% protein).

The dry weight of beef, broccoli, peanuts, and peas is about one-third protein, whereas cereals and fruits are less than 10% dry weight protein. Unlike many other plant proteins, legumes are not particularly low in lysine, and they are close to animal protein in threonine content. Vegetarians attempting to achieve complete protein often combine cereals (which are relatively high in methionine for plant protein) with legumes (which are relatively high in lysine for plant protein).

PHYTIC ACID
[PHYTIC ACID]

Lentils and other beans contain high amounts of phytic acid (phosphate-rich inositol), which can chelate positively-charged multivalent mineral ions (especially iron, zinc, magnesium, and calcium), preventing absorption. Soaking lentils and beans in warm water overnight not only makes them easier to cook, it allows some of the phytates to be soaked-out (and thrown-away with the water). Acidic solution (such as vinegar) better removes the phytates. Cooking also helps destroy phytates.

Although it would be very difficult to determine a diet providing optimum methionine for maximum human lifespan — even on the basis of rat experiments — evidence is convincing that reducing dietary methionine can help extend lifespan. The Table 2, listing milligrams of methionine per 100 grams of food (rather than per gram of protein, as in Table 1), could be helpful. Table values are based on [FOOD VALUES OF PORTIONS COMMONLY USED by Jean Pennington (1989)].

 

TABLE 2 Methionine in Foods
(milligrams/100 grams of food)
Food Methionine
Cheese, parmesan (dry) 971
Skim milk (dry) 907
Tuna (light) 862
Cheese, Swiss (processed) 792
Corned beef 711
Cheese, Cheddar 661
Salmon 631
Cheese, American (processed) 579
Extra lean beef 572
Walnuts, black 479
Egg white 394
Whole boiled egg 392
Pistashio nuts 386
Peanuts 289
Walnuts, Persian (English) 286
Cashew nuts 279
Cheerios 254
Oatmeal 250
Broad (Fava) beans 239
Soybeans 224
Barley 208
Tofu (firm) 202
Grape nuts (cereal) 200
Shredded wheat (cereal) 193
Wheaties (cereal) 168
Rice 167
Almonds 161
Yogurt 155
White beans 146
Black turtle beans 141
Navy beans 131
Kidney (red) beans 130
Chickpeas (garbanzos) 116
Blackeyed peas (cowpeas) 110
Lima beans 100
Macadamia nuts 93
Millet 85
Peas (raw) 82
Adzuki beans 79
Lentils 77
Corn 70
Spaghetti 51
Sweet potato (baked) 42
Mushrooms 40
Avacado 39
Mung beans 35
Broccoli 34
Potato 33
Pinto beans 33
Amaranth 30
Cauliflower 28
Oranges 22
Tomato paste 19
Kale 18
Banana 17
Blueberries 11
Onion 10
Tomato 8
Apple 2
Grapefruit 2
Strawberries 1

 

The absolute methionine content of a food is better evaluated knowing what the water, fat, carbohydrate, fiber, and protein content of that food is. A higher protein content and a lower methionine content is better than having a low methionine content because the food is low in protein and high in water, fat, or carbohydrate. Lima beans and rice are relatively high in both carbohydrate and methionine. Onions and strawberries are low in methionine, but are high in water and low in protein.

The data for Table 3 is taken from [NUTRITIVE VALUE OF FOODS; USDA Bulletin 72 (1981)], but is adjusted to give percent protein by dry weight. Percent water in the food is not related to the other columns. Fiber content is not given, and I suspect that fiber is equated with carbohydrate. I may have made a few errors, and I suspect that the data contains a few errors (garbage-in, garbage-out). But for the most part I think the data is good, my transcription is accurate, and my calculations are correct.

 

TABLE 3 Percent Macronutrients (dry weight)
and Percent Water (whole food)
Food Protein Carbohydrate Fat Water
Egg, white 100 0 0 88
Tuna solid,white, water 97 0 3 63
Salmon (baked) 81 0 19 67
Tuna chunk,light,oil 77 0 23 61
Corned beef 69 0 31 59
Ground beef,lean 57 0 43 56
Cheese, Parmesan (grated) 55 5 40 18
Ham 54 0 46 53
Ground beef,regular 53 0 47 54
Cheese, Swiss 47 7 47 42
Egg, whole 46 8 46 75
Yogurt, nonfat 43 57 0 80
Soybeans 41 20 39 71
Cheese, American processed 40 0 60 39
Milk, nonfat 39 59 2 91
Sesame seeds 29 14 57 5
Lentils, cooked 29 69 2 72
Sausage 29 0 71 45
Peas, split 27 71 2 70
Walnut, black 26 13 61 4
Frankfurter 26 5 68 54
Chickpeas (garbanzos) 23 75 6 60
Pinto beans, cooked 23 75 2 65
Pistachio nuts 22 26 52 4
Mushroom, cooked 25 67 8 91
Lima beans, cooked 24 74 2 64
Cashew nuts 16 35 49 2
Macaroni (enriched) 15 83 2 64
Tomato paste 16 80 3 74
Bread, whole wheat 16 76 7 38
Bread,1/3 wht (Pmnk) 16 78 6 37
Spaghetti (enriched), ckd 15 83 2 64
Egg noodles 15 80 4 70
Walnut, Persian (English) 15 19 65 4
Bread,2/3 wht (rye) 14 79 7 37
Onions 14 85 0 91
Corn 13 87 0 76
Potato (baked+skin) 9 91 0 71
Rice, brown 9 89 2 70
Avacado flesh (Florida) 8 46 46 80
Strawberries (raw) 8 83 8 92
Rice, white 7 93 0 73

 

Brown rice would be more nutritious than white rice, except that the fats in germ that is removed to make white rice can go rancid. Ingestion of Advanced Glycation End-Products (AGES) is detrimental to health.

Table 4gives the percent fat obtained for selected items in the above table, and breaks down the fat into percent saturated, monosaturated, and polyunsaturated fat. Numbers are rounded to the nearest whole number, which is why the total percentages don’t always add to 100. Monosaturated fats and polyunsaturated fats are preferred to unsaturated fats except where there is rancidity. Again, ingestion of Advanced Glycation End-Products (AGES) is detrimental to health. I had no data for non-fat cheese, the only kind of cheese that I eat.

 

TABLE 4 Percent Fat Types
(rounded)
Food Saturated Monosaturated Polyunsaturated % Fat
Cheese, American processed 67 30 4 52
Ground beef,lean 45 50 4 56
Egg, whole 44 52 4 48
Corned beef 44 52 4 29
Frankfurter 39 51 10 63
Ham 39 49 12 45
Sausage 37 50 11 69
Salmon (baked) 24 48 28 18
Tuna chunk,light,oil 22 30 48 18
Avacado flesh (Florida) 22 60 18 80
Cashew nuts 21 62 18 63
Soybeans 15 22 62 33
Pistachio nuts 13 71 16 52
Walnut, Persian (English) 9 24 66 64
Walnut, black 7 24 70 59

 

Table 5 gives relative proportions of all of the essential amino acids (plus tyrosine) for some representative high-protein animal foods as well as for some low-methionine plant foods.

Lysine is given after methionine because lysine is most often the limiting amino acid (the essential amino acid found in the smallest quantity relative to requirement) in cereals, nuts, and seeds — but lysine in abundant in legumes, for which methionine is typically the limiting amino acid [AMERICAN JOURNAL OF CLINICAL NUTRITION; Young,VR; 59(suppl):1203s-1212s (1994)]. Lysine is therefore listed second in the table. Leucine is listed third because of its paradoxical ability to reduce fat in high doses [DIABETES; Zhang,Y; 56(6):1647-1654 (2007)] and low doses [DIABETES; Cheng,Y; 59(1):17-25 (2010)]. Leucine and threonine are the limiting amino acid in vegetables and fruits, although vegetables and fruits are too low in protein to be considered significant proteins sources. Trytophan restriction has been shown to have a modest (compared to methionine restriction) ability to extend lifespan in rats [ MECHANISMS OF AGEING AND DEVELOPMENT; Ooka,H; 43(1):79-98 (1988)], reputedly by opposing an age-related increase in brain serotonin.

Tyramine was evaluated because of claims that high dietary tyramine could have adverse reactions with monoamine oxidase inhibitors (I take deprenyl). But none of the foods listed have seriously high levels of tyramine, so tyramine is not really a concern.

Again, this data is taken from  [FOOD VALUES OF PORTIONS COMMONLY USED by Jean Pennington (1989)]. I have adjusted the Pennington data to be standardized for 100 grams of food, rather than reproducing the variable quantities of food given, which makes comparison difficult. I may have made transcription errors, but probably not many (if any).

 

TABLE 5 Low Methionine Beans/Grains
Essential amino acids (+tyramine)
(milligrams/100 grams food)
Met = Methionine
Lys = Lysine
Leu = Leucine
Thr = Threonine
Try = Typtophan
Iso = Isoleucine
Phe = Phenylalanine
Val = Valine
His = Histidine
Tyr = Tyrosine
Food Met Lys Leu Thr Try Iso Phe Val His Tyr
Skim milk,dry 907 2867 3543 1633 510 2187 1746 2420 980 1747
American cheese 579 2225 1982 729 329 1036 1139 1343 914 1229
Walnuts, black 479 732 1729 739 325 993 1086 1304 489 761
Egg white 394 642 882 451 155 618 636 761 230 406
Walnuts, Persian (English) 286 293 1007 454 193 575 636 732 364 446
Cashew nuts 279 829 1304 600 239 743 804 1054 404 496
Soybeans, cooked 224 1108 1355 723 242 807 869 831 449 630
Whey, dry 200 967 1067 567 233 567 567 300 233 367
Rice, cooked 167 292 542 333 83 292 375 458 208 375
Yogurt, nonfat 169 514 578 235 32 312 312 474 142 289
Kidney (red) beans 130 595 693 365 103 383 469 454 241 244
Chickpeas (garbanzos) 116 593 631 329 85 380 475 372 244 220
Blackeyed peas (cowpeas) 110 523 592 294 95 314 451 368 240 250
Lima beans, cooked 100 523 673 337 92 411 470 469 238 276
Peas (raw) 82 317 323 203 37 195 200 235 106 113
Adzuki beans 79 567 632 255 72 300 398 387 198 224
Lentils, cooked 77 779 809 400 100 482 551 554 314 298
Corn, cooked 70 141 359 133 23 133 155 191 91 126
Broadbeans (Fava) 62 468 572 270 72 306 97 338 193 241
Spaghetti, cooked 51 109 220 133 41 170 177 _ 80 113
Mushrooms 40 211 129 94 46 82 80 97 57 46
Potato, baked 33 126 124 75 32 84 92 117 45 77
Pinto beans, cooked 33 564 656 346 97 363 444 430 229 231
Amaranth 30 109 167 85 27 102 114 118 44 68
Avacado flesh (Florida) 29 75 99 53 17 57 54 78 23 39
Mung beans, cooked 25 123 131 58 27 39 98 85 97 52
Tomato paste 19 108 105 86 26 73 80 77 60 51
Onion 10 56 41 28 18 42 30 28 19 29

 

Confusion can be caused by the variable amounts of proteins in the foods. Some foods have high water content (such as onion), or high carbohydrate content (such as rice), or high fat content (such as nuts). To compare relative amounts of methionine in the proteins in the foods, I have created Table 6 in which I have adjusted the values to reflect milligrams of amino acid per gram of protein, rather than the per 100 grams of food used in the previous table. To do this, I first calculate dry weight [(100 − % water) / 100] and then divide by % protein. (Note that Persian/English walnuts contain 60% the protein of black walnuts, mostly because of higher fat content. This creates a misimpression that Persian/English walnuts are much lower in methionine than black walnuts.)

I make no guarantee that I have made no transcription errors in manually copying data from either table to my calculator.

 

>

TABLE 6 Low Methionine Beans/Grains
Essential amino acids
(milligrams/gram protein)
Met = Methionine
Lys = Lysine
Leu = Leucine
Thr = Threonine
Try = Typtophan
Iso = Isoleucine
Phe = Phenylalanine
Val = Valine
His = Histidine
% P = % Protein (dry weight)
% W = % Water
Food Met Lys Leu Thr Try Iso Phe Val His % P % W
Rice (dry) 24 42 77 48 12 42 54 65 30 7 70
Milk,nonfat 24 75 92 43 13 57 45 63 26 40 91
Cheese, American 24 91 81 30 13 42 47 55 37 40 39
Corn, cooked 22 45 115 43 7.4 43 50 61 29 13 76
Sesame seeds 21 20 47 25 13 26 33 34 18 29 5
Walnuts, Persian (English) 20 20 70 32 13 40 44 51 25 15 4
Walnuts, black 19 29 69 30 13 40 44 52 20 26 4
Yogurt, skim 19 60 67 27 3.7 36 36 55 17 43 80
Soybeans, cooked 19 93 114 61 20 68 73 70 38 41 71
Cashew nuts 18 53 83 38 15 47 51 67 26 16 2
Avacado flesh (Florida) 18 47 62 33 11 36 34 49 14 8 80
Mushroom, cooked 18 94 57 42 20 36 36 45 25 25 91
Chickpeas (garbanzos) 13 64 69 36 9.2 41 52 40 27 23 60
Potato 13 48 48 29 12 32 35 45 17 9 71
Lentils, cooked 9.5 96 100 49 12 59 68 68 39 29 72
Spaghetti, cooked 9.4 20 41 25 7.6 31 33 _ 15 15 64
Onion 8 45 33 22 14 34 24 22 15 14 91
Tomato paste 4.5 26 25 21 6 18 19 19 14 16 74
Pinto beans, cooked 4 70 81 43 12 45 55 53 28 23 65

 

I am searching for foods that are high in protein, but low in methionine, as a source of protein. Preferably the foods should be high in the essential amino acids (other than methionine), and low in fat (especially saturated fat) and low in carbohydrate. As sources of protein, the data in the Table 6 are important in proportion to the percent protein in the food, especially when the water content is low. As long as protein is adequate in the diet overall, other foods that are low in protein and high in water are not much of a concern from a methionine restriction point of view. Legumes offer the best tradeoff of low methionine, and high protein (high essential amino acids), particularly lentils and pinto beans. Adzuki beans would be a contender except that the high fiber content makes them hard to process. I prefer to get my fiber from other sources.

Table 7 was created by dividing methionine amount into the amounts of the other essential amino acids shown in Table 5. Thus, the numbers in the lysine column reflect how many times the lysine content of the food exceed the methionine content.

 

TABLE 7 Ratio of Essential amino acids to methionine
(Essential amino acids)/methionine
Lys = Lysine
Leu = Leucine
Thr = Threonine
Try = Typtophan
Iso = Isoleucine
Phe = Phenylalanine
Val = Valine
His = Histidine
Food Lys Leu Thr Try Iso Phe Val His
Pinto beans, cooked 17 20 10 2.9 11 13 13 6.9
Lentils, cooked 10 11 5.2 1.3 6.3 7.2 7.2 4.1
Broadbeans (Fava) 7.5 9.2 4.3 1.2 6.4 1.6 5.5 3.1
Adzuki beans 7.2 8.0 3.2 0.91 3.8 5.0 4.9 2.5
Tomato paste 5.7 5.5 4.5 1.4 3.8 4.2 4.1 3.2
Onion 5.6 4.1 2.8 1.8 4.2 3.0 2.8 1.9
Mushrooms 5.3 3.2 2.6 1.2 2.0 2.0 2.4 1.4
Lima beans, cooked 5.2 6.7 3.4 0.92 4.1 4.7 4.7 2.4
Chickpeas (garbanzos) 5.1 5.4 2.8 0.73 3.3 4.1 3.2 2.1
Soybeans, cooked 4.9 6.0 3.2 1.1 3.6 3.9 3.7 2.0
Mung beans, cooked 4.9 5.2 2.3 1.1 1.6 3.9 3.4 3.9
Blackeyed peas (cowpeas) 4.8 5.4 2.7 0.86 2.9 4.1 3.3 2.2
Whey, dry 4.8 5.3 2.8 1.2 2.8 1.5 1.2 1.8
Kidney (red) beans 4.6 5.3 2.8 0.79 2.9 3.6 3.5 1.9
Peas (raw) 3.9 3.9 2.5 0.45 2.4 2.4 2.9 1.3
Potato, baked 3.8 3.8 2.3 0.97 2.5 2.8 3.5 1.4
Cheese, American 3.8 3.4 1.3 0.57 1.8 2.0 2.3 1.6
Amaranth 3.6 5.6 2.8 0.9 3.4 3.8 3.9 1.5
Skim milk,dry 3.2 3.9 1.8 0.56 2.4 1.9 2.7 1.1
Cashew nuts 3.0 4.7 2.2 0.86 2,7 2.9 3.8 1.4
Yogurt, nonfat 3.0 3.4 1.4 0.19 1.8 1.8 2.8 0.84
Avacado flesh (Florida) 2.6 3.4 1.8 0.59 2.0 1.9 2.7 0.79
Spaghetti, cooked 2.1 4.3 2.6 0.80 3.3 3.5 _ 1.6
Corn, cooked 2.0 5.1 1.9 0.32 1.9 2.2 2.7 1.3
Rice, cooked 1.7 3.2 2.0 0.50 1.7 2.2 2.7 1.2
Egg white 1.6 2.2 1.1 0.39 1.6 1.6 1.9 0.58
Walnuts, black 1.5 3.6 1.5 0.68 2.1 2.3 2.7 1.0
Walnuts, Persian (English) 1.0 3.5 1.6 0.67 2.0 2.2 2.6 1.3

 

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IV. METHIONINE RESTRICTION DIET

Pinto beans and lentils are the high-protein foods that show the best low-methionine, high-lysine profile, by a large margin. Lentils, however, are easier to soak before cooking to remove phytates, and produce a bit less odiferous flatulance than pinto beans. Both legumes, however, are high in phytic acid and raffinose oligosaccharides. Humans lack the enzyme to digest raffinose, which passes to the lower intestine where bacteria possessing the digestive enzyme create gases which can be quite odiferous.

Soaking pinto beans for 16 hours at room temperature only reduces raffinose oligosaccharides by 10%, and 90 minutes of cooling only cuts the raffinose oligosaccharide content in half [JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY; Song,D; 54(4):1296-1301 (2006)].

Just as the objective of calorie restriction is not to live without calories, methionine is an essential amino acid that can be reduced to 60% normal consumption to obtain most of the benefit [BIOGERONTOLOGY; Caro,P; 9(3):183-196 (2008)]. That dietary objective can be met without the need to consume legumes.

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Source: LIFE EXTENSION BENEFITS OF METHIONINE RESTRICTION

The idea of using the body’s immune system to fight cancer has been around for a century, but only in the past half a dozen years have dramatic breakthroughs begun rocking the medical world.

“That’s when the tsunami came,” says Drew Pardoll, director of the Bloomberg-Kimmel Institute for Cancer Immunology at Johns Hopkins University, and those advances are spawning hundreds of clinical trials nationwide, plus generating intense interest from patients, physicians and investors. Yet researchers remember the past anti-cancer efforts that fizzled after initially showing promise — which explains why most say daunting hurdles and years of perseverance are still ahead.

Here’s a primer about the new treatments and how they work:

What is cancer immunotherapy?

Immunotherapy is a significantly different approach from conventional treatments such as chemotherapy or radiation. The latter attack the malignancy itself, while immunotherapy aims to empower the immune system to kill it.

Because of the immune system’s unique power, says the nonprofit Cancer Research Institute, this therapy could prove a formidable weapon against many kinds of cancer and offer long-term protection with reduced side effects.

Which immunotherapies are sparking excitement? 

Two types of immunotherapy are drawing most of the interest: checkpoint inhibitors, which remove “brakes” from the immune system, allowing it to see and go after cancer; and CAR T-cell therapy, which involves a more customized attack.

“Checkpoint” inhibitors are designed to block the ability of certain proteins to blunt or weaken the response of the immune system so it can’t recognize and go after abnormal cells. In normal times, such checkpoint proteins keep the immune system from being too aggressive and damaging the body. But cancer sometimes hijacks them and uses them to suppress the immune system’s response to disease.

The Food and Drug Administration has cleared four checkpoint inhibitors for adults: Yervoy, also known as ipilimumab; Keytruda, or pembrolizumab; Opdivo, or nivolumab, and Tecentriq, or atezolizumab. The drugs are approved for malignancies including melanoma and Hodgkin lymphoma, as well as lung, kidney and bladder cancer. The treatments also are being tested in a wide range of other cancers.

Former president Jimmy Carter was treated with Keytruda, surgery and radiation for advanced melanoma last year. He announced in December that all signs of his cancer had disappeared.

In CAR T-cell therapy, T cells — a key part of the immune system — are removed from a patient, genetically modified in the lab to target a specific cancer and infused back into the person. This treatment, available only in clinical trials, is being tested mainly for leukemia and lymphoma. The Food and Drug Administration is likely to approve the first CAR T-cell treatment next year or in 2018.

Of these two immunotherapy approaches, most research and investor interest is focused on checkpoint inhibitors. That’s because they are off-the-shelf treatments that are much easier to administer than customized T-cell therapy, said Crystal Mackall, a former National Cancer Institute researcher who’s now leading immunotherapy trials for Stanford University School of Medicine.

What are some of the main challenges in immunotherapy?

Among the biggest challenges are increasing the response rate among patients and turning initial responses into long-lasting remissions. CAR T-cell therapy often produces a high remission rate in blood-disorder trials, but a significant percentage of patients relapse.

Checkpoint inhibitors induce responses — signaling a tumor has been shrunk or stabilized — in an average of just about 20 percent of patients, said oncologist Elizabeth Jaffee, the deputy director of the Sidney Kimmel Comprehensive Cancer Center at Hopkins. Researchers need to understand why only some cases and some cancers respond. Why, for example, the treatment benefits melanoma but not pancreatic cancer. They think the key to improving effectiveness will be coming up with combination treatments, as happened with AIDS. Jaffee points out that the tide was turned against that disease only after researchers figured out how to use a “cocktail” of medications to keep people with HIV from developing AIDS.

Nationwide, combination trials are testing the simultaneous use of two or more checkpoint inhibitors, a checkpoint inhibitor with a CAR T-cell therapy or an immunotherapy plus radiation and chemotherapy. But combining these can increase safety risks.

Jill O’Donnell-Tormey, chief executive of the Cancer Research Institute, said researchers also are trying to understand tumors’ “micro-environments,” which contain cells and other factors that appear to sometimes suppress the immune system’s response to cancer. The institute, along with the American Association for Cancer Research and two European groups, sponsored the three-day conference in New York.

What are immunotherapy’s downsides?

By revving up the immune system, immunotherapy can cause sometimes serious damage to healthy tissue and organs. Researchers are working on ways to limit or even reverse the potential toxicity, but much work needs to be done.

CAR T-cell therapy poses two types of safety risks. Almost all patients get sick with flu-like symptoms, including high fever and pain, a week or so after the treatment; some end up in intensive care. The treatment also can cause brain swelling that can be fatal.

Yet standard treatments have major side effects as well. Chemotherapy and radiation, when used for children with leukemia, can cause long-term problems such as secondary cancers, infertility and heart damage. In many ways, researchers say, immunotherapy is less toxic over the long term and might eventually be a good first-line alternative to chemo and radiation.

Immunotherapy can carry higher price tags. For example, Merck’s checkpoint inhibitor, Keytruda, costs about $150,000 a year. Once CAR T-cell therapies are approved by the Food and Drug Administration, they may cost hundreds of thousands of dollars a year, according to some analysts. If the treatments are used as directed by the agency, chances are good that insurance will pay for at least some of that.

Does immunotherapy work for children?

Immunotherapy in kids is a mixed picture.

Checkpoint inhibitors are only now being tested extensively in children, so it will take time to see how well they work. But very early-stage studies suggest that they may not be as effective as in adults. One theory holds that these drugs work better in cancers with many mutations — and pediatric cancers tend to have many fewer mutations.

CAR T-cell treatment, on the other hand, is being widely tested in children and has shown impressive effectiveness against acute lymphoblastic leukemia, the most common childhood leukemia.

How do I find immunotherapy treatments?

Talk first to your doctor, who should be able to help you find appropriate medication or clinical trials for unapproved treatment. Trials sponsored by the National Cancer Institute can be found at trials.cancer.gov. Studies also are listed on the website ClinicalTrials.gov –though that doesn’t signify government endorsement or approval. Another resource is the Cancer Research Institute’s Clinical Trial Finder. 

Read more:

Family hopes immunotherapy will save young girl with tumor

Long-term survival rates lengthen for melanoma patients on immunotherapy

Brain cancer replaces leukemia as leading cause of cancer deaths in children

While confident that immunotherapy will play an increasing role in cancer treatment, researchers must overcome some obstacles.

Source: Cancer immunotherapy is moving fast. Here’s what you need to know. – The Washington Post

Driven by technological progress, human life expectancy has increased greatly since the nineteenth century. Demographic evidence has revealed an ongoing reduction in old-age mortality and a rise of the maximum age at death, which may gradually extend human longevity. Together with observations that lifespan in various animal species is flexible and can be increased by genetic or pharmaceutical intervention, these results have led to suggestions that longevity may not be subject to strict, species-specific genetic constraints. Here, by analysing global demographic data, we show that improvements in survival with age tend to decline after age 100, and that the age at death of the world’s oldest person has not increased since the 1990s. Our results strongly suggest that the maximum lifespan of humans is fixed and subject to natural constraints.

Source: Evidence for a limit to human lifespan : Nature : Nature Research

Source: Next Big Future: Microbiome impacts tissue repair and regeneration

Source: Next Big Future: Ten percent have immune systems that ignore HIV and thus the immune system is saved and AIDS does not develop

A new study finds that exercise releases a hormone that helps the body shed fat and keeps it from forming.

researchers at the University of Florida Health has learned more about how the hormone irisin helps convert calorie-storing white fat cells into brown fat cells that burn energy.

The investigators also discovered that irisin—which typically surges when the heart and other muscles are exerted—inhibits the formation of fatty tissue, making it an attractive target for fighting obesity and diabetes. Irisin appears to work by boosting the activity of genes and a protein that are crucial to turning white fat cells into brown cells. Additionally, the researchers found that irisin significantly increases the amount of energy used by those cells, indicating it has a role in burning fat.

In the study, the research team collected fat cells donated by 28 patients who had breast reduction surgery. After exposing the samples to irisin, they found a fivefold increase in cells that contain a protein known as UCP1, or uncoupling protein 1, that is crucial to lipid metabolism.

“Human primary adipocytes derived from 28 female donors’ fresh subcutaneous white adipose tissue (scWAT) were used to examine the effects of irisin on browning and mitochondrial respiration, and preadipocytes were used to examine the effects of irisin on adipogenesis and osteogenesis,” the authors wrote. “Cultured fragments of scWAT and perirenal brown fat were used for investigating signal transduction pathways that mediate irisin’s browning effect by Western blotting to detect phosphorylated forms of p38, ERK, and STAT3 as well as uncoupling protein 1 (UCP1). Individual responses to irisin in scWAT were correlated with basal expression levels of brown/beige genes. Irisin up-regulated the expression of browning-associated genes and UCP1 protein in both cultured primary mature adipocytes and fresh adipose tissues.”

The findings from this study were published recently in the American Journal of Physiology–Endocrinology and Metabolism in an article entitled “Irisin Exerts Dual Effects on Browning and Adipogenesis of Human White Adipocytes.”

“We used human fat tissue cultures to prove that irisin has a positive effect by turning white fat into brown fat and that it increases the body’s fat-burning ability,” explained senior study investigator Li-Jun Yang, M.D., professor of hematopathology at the University of Florida’s College of Medicine’s department of pathology, immunology, and laboratory medicine.

Dr. Yang and her colleagues also found that irisin suppressed fat-cell formation. Among the tested fat tissue samples, irisin reduced the number of mature fat cells by 20–60% compared with those of a control group. That suggests irisin reduces fat storage in the body by hindering the process that turns undifferentiated stem cells into fat cells while also promoting the stem cells’ differentiation into bone-forming cells.

Because more than two-thirds of U.S. adults are overweight or obese, according to the National Institutes of Health, knowing that the body produces small quantities of fat-fighting irisin highlights the importance of regular exercise.

“Instead of waiting for a miracle drug, you can help yourself by changing your lifestyle,” Dr. Yang stressed. “Exercise produces more irisin, which has many beneficial effects including fat reduction, stronger bones, and better cardiovascular health. Irisin can do a lot of things, and this is another piece of evidence about the mechanisms that prevent fat buildup and promote the development of healthy bones when you exercise.”

Source: Exercise Hormone has Dual Role in Shedding and Preventing Fat | GEN News Highlights | GEN

Sex doesn’t end as you age, so keep on using “it.”

Source: Use It Or Lose It: How Age, Hormones, And Masturbation Predict Sexual Health

After three months, more than a third of study participants grew back more than half of their lost hair

Source: Arthritis drug Xeljanz may help with the hair loss condition alopecia – CBS News

Euthanasia is on it’s way

Treating a seriously ill patient who suffers from multiple chronic conditions can be difficult and expensive. These so-called high-need, high-cost (HNHC), or “complex care” patients make up about 5 percent of the U.S. population, but by some estimates, account for 50 percent of healthcare spending.

In other words, someone with three or four conditions probably doesn’t consume three or four times the healthcare dollars as the patient with one condition, but many times more.

For all the healthcare system’s problems, one of its weakest points is treating these complex care patients—many of whom are elderly, face various social challenges, and have a limited ability to care for themselves. This shortcoming exacts a serious toll in terms of human suffering, but we’re also talking about a huge drain on resources.

“Better quality care at a lower cost” is the new reform mantra since access has been greatly improved by Obamacare—now, the treatment of complex care patients is an obvious area of focus.

Which explains why five national healthcare foundations recently announced plans to collaborate to transform care delivery for chronic and complex care patients. The groups—the Commonwealth Fund, the John A. Hartford Foundation, Robert Wood Johnson Foundation, the Peterson Center on Healthcare and the SCAN Foundation—said they would start work later this year.

Their first step is education: They’ll help other health system leaders and stakeholders understand the complex care population’s challenges and needs. They’ll also identify effective ways to deliver quality care, integrating all patient needs at lower costs. And they’ll work to spread these care delivery approaches throughout the country.

This isn’t new terrain for healthcare funders, as we’ve reported before. But this new collaboration is significant. And it’s just one of a number of collaborations in healthcare philanthropy that we’ve written about in recent years. Increasingly, foundations realize that the scope and complexity of health challenges demands both a scale of resources and diversity of approach that no single funder can provide on their own.

Related: 

The partners in this collaborative outlined the problem and their goals in an article published in the New England Journal of Medicine. “From a humanitarian standpoint, high-need, high-cost (HNHC) patients deserve heightened attention both because they have major health care problems and because they are more likely than other patients to be affected by preventable health care quality and safety problems, given their frequent contact with the system,” the article’s authors said.

Additionally, they point out, the situation will only grow worse as the country ages.

Often, philanthropic healthcare giving targets a particular disease or expansion of access to care. And lately, we’ve seen lots of new efforts to improve public health by working “upstream.” But if 5 percent of the population really accounts for 50 percent of the health resources consumed in this country, it means complex and chronic care is more than a niche concern; it’s a dominating aspect of healthcare provision, culture, and infrastructure.

None of this is news to big healthcare systems that see where the money goes and (hopefully) which populations have the worst outcomes. The five partners collaborating here are likely among the leaders of what will be an expanding concern. Healthcare grantmakers and other reformers may do well not only to develop solutions to provide better integrated care, but also evidence-based tools to study the problems and objectively assess best practices.

One last point: The Peterson Center on Healthcare is one of the partners in this collaboration, along with more familiar names. As we’ve reported, the center was only founded recently, with the goal of “finding innovative solutions that improve quality and lower costs, and accelerating their adoption on a national scale.” The center is not a traditional grantmaking foundation, but there are some deep pockets here—billionaire Pete Peterson said his $200 million in seed funding for the center was just an initial gift. So it’s worth watch closely as this new player gets fully up and running.

Source: The Elephant in the Waiting Room: Behind a New Healthcare Collaborative  – Inside Philanthropy – Inside Philanthropy

INFLAMMATION: The Cardiac Killer

Citations

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Barter P, Gotto AM, LaRosa JC, Maroni J, Szarek M, Grundy SM, Kastelein JJ, Bittner V, Fruchart JC; Treating to New Targets Investigators. HDL cholesterol, very low levels of LDL cholesterol, and cardiovascular events. N Engl J Med. 2007 Sep 27;357(13):1301-10. [Link]

Cabrera MAS, de Andrade SM, Dip RM. Lipids and All-Cause Mortality among Older Adults: A 12-Year Follow-Up Study. The Scientific World Journal. 2012;2012:930139. doi:10.1100/2012/930139. [Link]

Ridker PM, Danielson E, Fonseca FA, Genest J, Gotto AM Jr, Kastelein JJ, Koenig W, Libby P, Lorenzatti AJ, MacFadyen JG, Nordestgaard BG, Shepherd J, Willerson JT, Glynn RJ; JUPITER Study Group. Rosuvastatin to prevent vascular events in men and women with elevated C-reactive protein. N Engl J Med. 2008 Nov 20;359(21):2195-207. [Link]

C-Reactive Protein, Fibrinogen, and Cardiovascular Disease Prediction The Emerging Risk Factors Collaboration. N Engl J Med 2012; 367:1310-1320October 4, 2012DOI: 10.1056/NEJMoa1107477 [Link]

Rucker R, Chowanadisai W, Nakano M. Potential physiological importance of pyrroloquinoline quinone. Altern Med Rev. 2009 Sep;14(3):268-77. [Link]

W, Bauerly K, Tchaparian E, Rucker RB. Pyrroloquinoline quinone (PQQ) stimulates mitochondrial biogenesis. FASEB J 2007;21:854 [Link]

Ohwada K, Takeda H, Yamazaki M, et al. Pyrroloquinoline quinone (PQQ) prevents cognitive deficit caused by oxidative stress in rats. J Clin Biochem Nutr 2008;42:29-34. [Link]

Bauerly K, Harris C, Chowanadisai W, et al. Altering Pyrroloquinoline Quinone Nutritional Status Modulates Mitochondrial, Lipid, and Energy Metabolism in Rats. Hansen IA, ed. PLoS ONE. 2011;6(7):e21779. doi:10.1371/journal.pone.0021779. [Link]

Harris, Calliandra B. et al. Dietary pyrroloquinoline quinone (PQQ) alters indicators of inflammation and mitochondrial-related metabolism in human subjects. Journal of Nutritional Biochemistry , Volume 24 , Issue 12 , 2076 – 2084 [Link]

Windler E, Schöffauer M, Zyriax BC. The significance of low HDL-cholesterol levels in an ageing society at increased risk for cardiovascular disease. Diab Vasc Dis Res. 2007 Jun;4(2):136-42. [Link]

Lee S Gross, Li Li, Earl S Ford, and Simin Liu. Increased consumption of refined carbohydrates and the epidemic of type 2 diabetes in the United States: an ecologic assessment. Am J Clin Nutr May 2004 vol. 79 no. 5 774-779 [Link]

López-Alarcón M, Perichart-Perera O, Flores-Huerta S, et al. Excessive Refined Carbohydrates and Scarce Micronutrients Intakes Increase Inflammatory Mediators and Insulin Resistance in Prepubertal and Pubertal Obese Children Independently of Obesity. Mediators of Inflammation. 2014;2014:849031. doi:10.1155/2014/849031. [Link]

Siri-Tarino PW, Sun Q, Hu FB, Krauss RM. Meta-analysis of prospective cohort studies evaluating the association of saturated fat with cardiovascular disease. The American Journal of Clinical Nutrition. 2010;91(3):535-546. doi:10.3945/ajcn.2009.27725. [Link]

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Koh AS, Simmons-Willis TA, Pritchard JB, Grassl SM, Ballatori N. Identification of a mechanism by which the methylmercury antidotes N-acetylcysteine and dimercaptopropanesulfonate enhance urinary metal excretion: transport by the renal organic anion transporter-1. Mol Pharmacol 2002 Oct;62(4):921-6 [Link]

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Source: The Cardiac Killer

Autophagy – the housekeeper in every cell that fights aging

By James P Watson and Vince Giuliano

Background and introduction

There is a wide variety of genetic manipulations, pharmacologic manipulations, and nutrient manipulations that have been shown to alter lifespan in model organisms.  These include caloric restriction, “loss of function” mutations, “gene knock out” models, phytochemicals, and drugs that down regulate aging pathways (mTOR, insulin/IGF-1, etc.).  It also includes “gain of function mutations”, transgenic models, phytochemicals, and drugs that up regulate longevity promoting pathways (AMPK, FOXO, Klotho, etc.).  At first glance, all these interventions may seem to be unrelated, suggesting that aging is a multifactorial problem with no common denominator to longevity.  On further examination, however, there is a common denominator to all of these interventions – autophagy.  Autophagy (“self eating”) is an old, evolutionarily conserved stress response that is present in all living cells. Like apoptosis, autophagy is a programmed response and has several sub-pathways.  Unlike apoptosis, autophagy promotes life rather than death.  Recent discoveries have shown that almost every genetic, dietary, and pharmacologic manipulation proven to extend lifespan activates autophagy as part of its mechanism of action.

Autophagy is the way your cells “clean house” and “recycle the trash”.  Along with the ubiquitin proteasome system, autophagy is one of the main methods that cells use to clear dysfunctional or misfolded proteins.  Autophagy can clear any kind of trash: intracellular viruses, bacteria, damaged proteins, protein aggregates and subcellular organelles. Although autophagy has long been known to exist, only recently has there been a clear understanding of the genes and pathways related to it.  This recent evidence suggests that the declining efficacy of autophagy may be a driver of many of the phenotypic phenomena of aging.  This blog entry explores the “evidence for the autophagy theory of aging” and builds a strong case that defective autophagy is a central driver for age-related diseases and aging itself.

Autophagy now appears to be a downstream event following insulin/IGF-1 pathway down-regulation, mTOR inhibition, Klotho activation, AMPK activation, Sirtuin dependent protein deacetylation, and histone acetyl transferase inhibition.  Autophagy explains in part, the beneficial effects of caloric restriction, caffeine, green tea, rapamycin, resveratrol, metformin, spermidine, lithium, exercise, hypoxia, Torin-1, trehalose, and a host of other natural and synthetic compounds.

There is much stronger evidence of a link between autophagy activation and longevity than there is with any other longevity interventions such as exogenous anti-oxidant supplementation, endogenous anti-oxidant up regulation, micronutrient replacement, hormone replacement, anti-inflammatory therapy, telomerase activation, or stem cell therapy.   For this reason, we have listed below the top reasons why “eating yourself for dinner” mauy well be the best way to promote health and longevity.

What is autophagy?

Biological entities employ various mechanisms to keep themselves functioning healthily, including mechanisms to get rid of defective or no longer wanted components.  Inter and intra-cell signaling can drive a cell to destroy itself, for example (cell apoptosis).  Short of apoptosis, on the cell level there are several mechanisms for getting rid of defective or no longer needed components including organelles and proteins.  From the 2008 publication Autophagy and aging:  “All cells rely on surveillance mechanisms, chaperones and proteolytic systems to control the quality of their proteins and organelles and to guarantee that any malfunctioning or damaged intracellular components are repaired or eliminated [1,2]. Molecular chaperones interact with unfolded or misfolded proteins and assist in their folding [3]. However, if the extent of protein damage is too great, or the cellular conditions are not adequate for re-folding, the same molecular chaperones often deliver proteins for degradation. Two proteolytic systems contribute to cellular clearance: the ubiquitin-proteasome and the lysosomal systems [4].”  Autophagy is concerned with the lysosomal system and involves the “degradation of any type of intracellular components including protein, organelles or any type of particulate structures (e.g. protein aggregates, cellular inclusions, etc.) in lysosomes(ref)”

process-of-autophagy

Image source

Autophagy, or autophagocytosis, is a catabolic process involving the degradation of a cell’s own components through the lysosomal machinery. It is a tightly regulated process that plays a normal part in cell growth, development, and homeostasis, helping to maintain a balance between the synthesis, degradation, and subsequent recycling of cellular products. It is a major mechanism by which a starving cell reallocates nutrients from unnecessary processes to more-essential processes. Autophagy is an evolutionarily conserved mechanism of cellular self-digestion in which proteins and organelles are degraded through delivery to lysosomes. Defects in this process are implicated in numerous human diseases including cancer(ref).”

Top 16 Key Facts about Autophagy

There are three main pathways of Autophagy – Macroautophagy, Microautophagy, and Chaperone-mediated Autophagy (CMA).

All 3 autophagy pathways are constitutively active (i.e. they can occur at basal levels) but can also be up regulated by cellular stress). Macroautophagy is the primary “broom” that sweeps the house. Macroautophagy is initiated when the material to be removed is tagged with ubiquitin.  This signals a complex series of molecular events that leads to the formation of a membrane  around the material to be removed and recycled.  This membrane formation around the debris is called a autophagosome.  Once formed, the autophagocome fuses with a lysosome to form an autolysosome.  Once fusion occurs, the acid hydrolases found inside the lysosomes start digesting the damaged proteins and organelles.  When damaged mitochondria are digested by macroautophagy, it is called mitophagy, which is a specific type of macroautophagy. Macro-autophagy can also remove and recycle mutated or free-radical damaged proteins or protein aggregates.  Macroautophagy  and other sub cellular organelles (peroxisomes, endoplasmic reticulum, etc.)  Even part of the cell nucleus can undergo autophagy (called “piecemeal microautophagy of the nucleus” – PMN).

Macroautophagy   Image source

macroautophagy

Chaperone-mediated autophagy (CMA) is a specific mechanism of autophagy that requires protein unfolding by chaperones.   The other two mechanisms do not require protein unfolding (macroautophagy and microautophagy).  Since protein aggregates cannot be unfolded by chaperone proteins, both the ubiquitin-proteasome system and chaperone-mediated autophagy are unable to clear these protein aggregates.  For this reason, macroautophagy may be the most important pathway for preventing Alzheimer’s disease, Parkinson’s disease, Fronto-temporal dementia, and all of the other neurodegenerative diseases associated with protein aggregate accumulation.

Microautophagy is essentially just an invagination (folding in) of the lysosomal membrane and does not require the formation of an double-membrane autophagosome.  Both CMA and microautophagy appear to play a minor role in “house keeping”.  Here are diagrams of these types of autophagy.

kindsofautophagy1

Image source

 

Image sourcekindsofautophagy

 2. Autophagy is the only way to Get Rid of Old Engines  i.e. damaged mitochondria

Autophagy is the best way to get rid of bad mitochondria without killing the cell.  The process is called “mitophagy.” Since bad mitochondria produce most of the “supra-hormetic doses of ROS”, this is really, really, important. This is explained in our recent blog entries related to mitochondria, Part 1, and Part 2.  For brain cells, heart cells, and other post mitotic cells that we all want to “hang on to”, mitophagy is probably the most important anti-aging value of mitophagy.  Bad mitochondria are phosphorylated by the kinase PINK1.  Then these bad mitochondria are ubiquinated by the E3 ligase Parkin.  The ubiquinated bad mitochondria are then selectively destroyed by mitophagy, which is a form of macroautophagy.

mitophagy1Mitophagy   Image source

The 2007 publication Selective degradation of mitochondria by mitophagy reviews the topic.  “Mitochondria are the essential site of aerobic energy production in eukaryotic cells. Reactive oxygen species (ROS) are an inevitable by-product of mitochondrial metabolism and can cause mitochondrial DNA mutations and dysfunction. Mitochondrial damage can also be the consequence of disease processes. Therefore, maintaining a healthy population of mitochondria is essential to the well-being of cells. Autophagic delivery to lysosomes is the major degradative pathway in mitochondrial turnover, and we use the term mitophagy to refer to mitochondrial degradation by autophagy. Although long assumed to be a random process, increasing evidence indicates that mitophagy is a selective process.”

3. Autophagy is the best Way to Get Rid of Junk.    – protein aggregates, etc.

Autophagy is the best way to get rid of protein aggregates like those associated with all of the neurodegenerative diseases, like amyloid beta, tau tangles, alpha synuclein aggregates, TDP-43 aggregates, SOD aggregates, and Huntington protein aggregates.  These aggregates are NOT digested via the ubiquitin-proteasome system, since they cannot be “unfolded”.   For this reason, autophagy is probably the most important cellular mechanism for clearing protein aggregates found in neurodegenerative diseases.  Autophagy can also clear out bad cytoplasm (Cvt), endoplasmic reticulum, peroxisomes (micro and macropexophagy), Golgi apparatus,  and even damaged parts of the nucleus (PMN).  See for example (2012) Degradation of tau protein by autophagy and proteasomal pathways and (2009) Autophagy protects neuron from Abeta-induced cytotoxicity

Autophagy is protective by quietly getting rid of multiple other unwanted substances.  For example, it protects against alcohol-induced liver damage.  Consider what is going on in this diagram from the 2011 publication The emerging role of autophagy in alcoholic liver disease:

alcoholmitophagyImage source     “Alcohol consumption causes hepatic metabolic changes, oxidative stress, accumulation of lipid droplets and damaged mitochondria; all of these can be regulated by autophagy. This review summarizes the recent findings about the role and mechanisms of autophagy in alcoholic liver disease (ALD), and the possible intervention for treating ALD by modulating autophagy(ref).”

4. Aging = Autophagy decline. 

According to the 2008 publication Autophagy in aging and in neurodegenerative disorders: “Growing evidence has indicated that diminished autophagic activity may play a pivotal role in the aging process. Cellular aging is characterized by a progressive accumulation of non-functional cellular components owing to oxidative damage and a decline in turnover rate and housekeeping mechanisms. Lysosomes are key organelles in the aging process due to their involvement in both macroautophagy and other housekeeping mechanisms. Autophagosomes themselves have limited degrading capacity and rely on fusion with lysosomes. Accumulation of defective mitochondria also appears to be critical in the progression of aging. Inefficient removal of nonfunctional mitochondria by lysosomes constitutes a major issue in the aging process. Autophagy has been associated with a growing number of pathological conditions, including cancer, myopathies, and neurodegenerative disorders.”

The relationship of autophagy decline to hallmarks of aging has been known for a long time and have been best studied in liver cells.  The auto florescent protein lipofuscin is the oldest and simplest biomarker of declining autophagy and represents undigested material inside of cells.  The Lewy bodies seen in several neurodegenerative diseases (including “Parkinson’s disease with dementia”) are also biomarkers of declining autophagy and may specifically be due to “declining mitophagy”.  Declining autophagy is particularly important in post-mitotic cells such as those in the brain, heart, and skeletal muscle where very little cell regeneration via stem cells occurs.  For mitotic tissues such as the GI tract, bone marrow, and skin, autophagy decline may not be as detrimental, since apoptosis is another normal method for getting rid of bad cells.

The failure of autophagy with aging has several possible causes:

a. Fusion problems – Autophagic vacuoles accumulate with age in the liver.  This may be due to a problem of fusion between the autophagosomes and the lysosomes.

b. Glucagon deficiency – Glucagon is a hormone that enhances macroautophagy. “—the stimulatory effect of glucagon [on autophagy] is no longer observed in old animals.  See item (b) in the next list below.(ref)“

c. Negative signaling via the Insulin receptor – Insulin activates the Insulin/IGF-1 pathway which activates mTOR.  mTOR activation inhibits autophagy (see below).  Even in the absence of insulin, there is up-regulation with aging of the insulin/IGF-1 signaling via the insulin receptor tyrosine kinase.  This would activate mTOR.

d. Inadequate turnover of damaged mitochondria – Mitophagy decline may be one of the mechanisms that is responsible for the decline in autophagy with aging.  Specifically, if mitophagy does not keep up with the demand for damaged mitochondrial clearance, a higher baseline ROS would occur, which would damage proteins, cell membrane lipids, and cell nucleus DNA.

e. Energy compromise – With aging, there is a decline in energy production by the cells.  This may be one of the reasons for the decline in autophagy seen in aging.

Here is a depiction of some of the main problems associated with decline of autophagy in aging:

conseqagingautop

Some consequences of failure of autophagy with aging  “Possible causes and consequences of the failure of macroautophagy in old organisms are depicted in this schematic model (brown boxes”   Image source

(a) The accumulation of autophagic vacuoles with age could result from the inability of

lipofuscin- loaded lysosomes to fuse with autophagic vacuoles and degrade the sequestered content.

(b) In addition, the formation of autophagosomes in old cells might be reduced because of the inability of macroautophagy enhancers (such as glucagon) to induce full activation of this pathway. The stimulatory effect of glucagon is compromised in old cells because of maintained negative signaling through the insulin receptor (IR) even under basal conditions (i.e. in the absence of insulin). Maintained insulin signaling would activate mTOR, a known repressor of macroautophagy.

(c) Inadequate turnover of organelles, such as mitochondria, in aging cells could increase levels of free radicals that generate protein damage and

(d) Aging could also potentiate the inhibitory signaling through the insulin receptor.

(e) An age-dependent decline in macroautophagy can also result in energetic compromise of the aging cells.

5.  Genetic manipulations that increase lifespan in all model organisms stimulate autophagy.

Knocking out macroautophagy takes away at least 50% of the long-lived mutant’s added lifespan.  This same “loss of longevity” is seen with Caloric restriction in “macroautophagy knockouts”.    The following diagram shows how important autophagy is in long-lived mutant nematodes and how this is important for increasing lifespan, reducing cellular damage, and increasing function.

autophagymutants

Image source

The most well studied “mutants” are model organisms where one of the following pathways are altered by a gene mutation or a gene knock out.  When an additional “knocking out” of an autophagy gene is done, approximately 1/2 of the added lifespan of the long lived mutants (vs wild type) appears to be “wiped out” by loosing autophagy.   Similar findings occur in “macroautophagy  knock-outs” subjected to caloric restriction, etc.  This suggests to me that 1/2 of the benefits of caloric restriction are due to stimulating autophagy.  Caloric restriction down regulates all of the”nutrient sensing pathways that are negative regulators of autophagy” and up regulates other “ nutrient sensing pathways that are positive regulators of autophagy”.  The following interconnected “nutrient -sensing pathways” affect macroautophagy:

a. IGF-1: two mechanisms:

i. decreasing Insulin-IGF-1 pathway => tyrosine kinase => inhibits Akt phosphorylation of TSC =>  inhibition of raptor in mTOR complex

ii. decreasing insulin/IGF-1 pathway => Foxo transcription factor translocation to nucleus  => FOXO stimulates autophagy via activating two  autophagy genes – LC3 and BNIP3.

b. mTOR:  three mechanisms account for the activation of autophagy by mTOR inhibition

i.  mTOR inhibition => decreases phosphorylation of Atg1 (aka ULK1/2). Also decreases phosphorylation of  Atg13 and Atg17.  Phosphorylation of ULK1/2, Atg13, and Atg17 inhibits autophagy initiation.

ii. decreasing mTOR pathway => decreases phosphorylation of 4EBP1 => blocks effect of eIF4F => autophagy activation.

iii. decreasing mTOR pathway => decreases phosphorylation of S6K => S6K no longer active => inhibition of autophagy.

Microsoft PowerPoint - Final IBDMN Fig 2

Signaling pathways that affect autophagy Image source

“The (mammalian) target of rapamycin (mTOR) is a primordial negative regulator of autophagy inorganisms from yeast to man. mTOR is inhibited under starvation conditions, and this contributes to starvation-induced autophagy via activation of mTOR targets Atg13, ULK1, and ULK2. This inhibition can be mimicked by mTOR inhibitory drugs like rapamycin (Ravikumar et al., 2010).  One of the important pathways regulating mTOR is initiated when growth factors like insulin-like growth factor bind to insulin-like growth factor receptors (IGF1R) (Figure 2). These receptors signal, via their tyrosine kinase activities, to effectors like the insulin receptor substrates (IRS1 and IRS2), which in turn activate Akt. Akt inhibits the activity of the TSC1/TSC2 (proteins mutated in tuberous sclerosis) complex, a negative regulator of mTOR. In this way, IGF1R signaling activates mTOR and inhibits autophagy, and the converse occurs when nutrients are depleted(ref).”

c. Ras/PKA:  decreasing Protein Kinase A pathway (aka Ras/cAMP) => decreases phosphorylation of 3 autophagy proteins (Atg1, Atg13, Atg18).

d. PKB/Akt: decreasing Protein Kinase B pathway (aka PkB/Akt or Sch9) => reduces inhibition of TSC-1 => decreased mTOR activity.

e. Sirtuin 1:  CR activates Sirtuin 1 => deacetylation of several autophagy gene products: Atg5, Atg7, Atg8/LC3.   Sirt1 also activates AMPK, activates FOXO3a, and inhibits mTOR via TSC-1/2

f. AMPK: AMPK pathway (aka LKB1-AMPK) activates autophagy via two methods:

i. AMPK activation => phosphorylates TSC2 and raptor => inhibits TORC1  (this requires glucose starvation).

ii. AMPK activation => direct phosphorylation of Atg1 (aka ULK1) => autophagy activation (this does NOT require glucose starvation).

g. Less-important pathways:

i.  Rim15:  increasing Rim15 Kinase pathway => Msn2 and Msn4 transcription factor translocation to nucleus => inhibits mTOR, PKA, and PKB pathways.

ii  ERK1/2:  ERK pathway – the extracellular signal-regulated kinase (ERK) also mediates starvation-induced autophagy.  (see #6 below for more details)

iii. JNK: JNK pathway – This is a MAPK that mediates starvation-induced autophagy. (see #6 below for more details).

The main pathways are depicted in the following diagram of how Calorie Restriction works (Ras/PKA and less important pathways not depicted).

Kroemer_3

Autophagy regulation      Image source

6. There are many other pathways that regulate autophagy that are not dependent on “nutrient sensing pathways.” 

(i.e. not those described above).

Although caloric restriction or fasting are clearly the most “potent” autophagy stimulators, since they can activate macroautophagy via the above “nutrient sensing pathwaysthere are many other pathways that can activate autophagy.  Here an explanation of the roles of the key kianses involved:

a. PI3Ks and Akt – PI3Ks are kinases that are mainly activated by growth factors, not starvation.  There are 3 classes of PI3Ks and the Class III PI3Ks directly positively activate autophagy (Vps34) whereas the Class I PI3Ks indirectly inhibit autophagy via mTOR and Akt.

b. MAPKs – Mitogen-Activated Protein Kinase – these are kinases that are mainly activated by growth factors, not starvation.  There are 3 classes:

i. ERK – Extracellular signal-Regulated Kinases (ERK) positively regulate autophagy by maturing autophagic vacuoles.  EKR also seems to specifically be involved with mitochondrial-specific autophagy (i.e. mitophagy).  Mitochondrial ERK may help protect from neurodegenerative diseases.  Cancer cells also activate mitochondrial ERK to cause chemoresistance.  ERK is activated downstream from Ras.  Ras activates Raf, which activates MEK.  MEK phosphorylates and activates ERK1 and ERK2.

This is the mechanism by which you can kill cancer with soy extracts, capsaicin, and Cadmium.  Here is how this works:

  • Soyasaponins (found in soybeans) => activates ERK => autophagy-induced death in colon cancer cells
  • Capsaicin (found in chili peppers) => activates ERK => autophagy-induced death in breast cancer cells
  • Cadmium (toxic metal) => activates ERK => autophagy-induced death in mesangial cells

ii. p38 – p38 is a MAPK that is a tumor suppressor.  p38 regulates autophagy but there is still controversy if it activates or inhibits autophagy.

iii. JNK – JNK is a MAPK that is activated by heat shock, osmotic shock, UV light, cytokines, starvation, T-cell receptor activation, neuronal excitotoxic stimulation, and ER stress.  With starvation, JNK does not phosphorylate Bcl-2, which prevents it from binding to beclin 1.  Beclin 1 can then induce autophagy.  Bcl-2 is an anti-apoptotic protein and can prevent apoptosis.  There are multiple phosphorylation sites on Bcl-2.  The degree by which JNK phosphorylates/dephosphorylates Bcl-2 may determine cell fate – i.e. apoptosis (death) vs autophagy (survival). See (2011) The Beclin 1 network regulates autophagy and apoptosis.

c. PKC – Protein Kinase C (PKC) is a family of kinases that were once thought to be associated mostly with apoptosis/anti-apototis.  Recent research has shown that PKCs also play a role in autophagy.  The effects of PKC depend on if the cellular stress is acute or chronic.  For instance, PKCg is an example of one of the PKCs where it stimulates autophagy with acute, short periods of hypoxia (via JNK activation) but suppresses autophagy with chronic hypoxia (via Caspace-3).   Another PKC, PKC0  is involved with ER-stress induced autophagy.  Acadesine (AICAR) induces autophagy via a PKC/Raf1/JNK pathway.  Acadesine (AICAR) in combination with GW1516 has shown to improve endurance-type exercise by converting fast-twitch muscle fibers into the more energy-efficient, fat-burning, slow-twitch muscle fibers.  These two compounds turned on 40% of the genes that were turned on when exercise + GW1516 were used together.  For this reason, acadesine (AICAR) has been termed an “exercise mimetic” and has been banned for use by athletes, since it is a performance enhancing drug, even though it is very safe.  The mechanism of action of AICAR may be in part its induction of autophagy.

d. Endoplasmic Reticulum Stress Kinases (i.e. the ER unfolded protein response) – Several kinases involved with the endoplasmic reticulum unfolded protein response (ER-UPR) have been found to activate autophagy.  They include the following:

i. IRE-1 – Inositol-requiring enzyme (IRE1) is one of the first proteins activated by the ER-UPR.  It up regulates autophagy genes (Atg5, 7, 8, 19).

ii. PERK – PERK must phosphorylate the eukaryotic initiation factor 2alpha (eIF2alpha) for LC3 conversion with ER-UPR induced autophagy.     PERK also up regulates Atg5.

iii. CaMKKbeta – ER stress results in calcium release from the ER.  This Ca++ release induces autophagy via the Ca dependent kinases.  The main one is called Ca/Calmodulin-dependent kinase beta (CaMKKbeta).  This is an “upstream activator” of AMPK, which in turn inhibits mTOR.  This is how calcium can induce autophagy.

iv. DAPK1 – Death-associated protein kinase 1 (DAPK1) is another Ca++/Calmodulin-regulated kinase that is important in ER-UPR induced autophagy. It induces autophagy by phosphorylating beclin 1, which is necessary for autophagosome formation.

erstressautophagy

Mechanisms connecting  ER stress and autophagyImage Source  “Mechanisms connecting ER stress and autophagy. Different ER stresses lead to autophagy activation. Ca2+ release from the ER can stimulate different kinases that regulate autophagy. CaCMKK phosphorylates and activates AMPK which leads to mTORC1 inhibition; DAPK phosphorylates Beclin-1 promoting its dissociation from Bcl-2; PKCθ activation may also promote autophagy independently of mTORC1. Inositol 1,4,5-trisphosphate receptor (IP3R) interacts with Beclin-1. Pharmacological inhibition of IP3R may lead to autophagy in a -independent manner by stimulating its dissociation from Beclin-1. The IRE1 arm of ER stress leads to JNK activation and increased phosphorylation of Bcl-2 which promotes its dissociation from Beclin-1. Increased phosphorylation of eIF2 in response to different ER stress stimuli can lead to autophagy through ATF4-dependent increased expression of Atg12. Alternatively, ATF4 and the stress-regulated protein p8 promote the up-regulation of the pseudokinase TRB3 which leads to inhibition of the Akt/mTORC1 axis to stimulate autophagy(ref).”

7. Excess baseline ROS from bad mitochondria induces Mitophagy.

 – ROS induces autophagy via a non-canonical pathway

This may be the mitochondrial signal for “selective destruction” of damaged mitochondria.  Exogenous ROS can also induce autophagy, however.  For instance, there is evidence that abnormal levels of H202 in the cytoplasm will induce macroautophagy. Hydrogen peroxide induces a “non-canonical autophagy” that is “beclin-1 independent” but requires the JNK-mediated activation of Atg7.  on of Atg7.

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ROS induces autophagy: Roles of Akt, ERK, JNK and BeclinsImage source

8. Most all of the Pharmacologic manipulations that extend lifespan increase autophagy.

Here are some of the main ones:

a. Rapamycin – Autophagy explains most of the longevity and health benefits (mechanism of action) of Rapamycin

Since the protein kinase mTOR phosphorylates the 3 key autophagy initiating proteins (Atg1, Atg13, and Atg17),  it is considered the  “Master of Autophagy”.  Rapamycin inhibits both TORC1 and TORC2.  TORC1 inhibition is the the “direct” and primary mechanism by which rapamycin activates autophagy, but TORC2 inhibition has an “indirect” and independent method of activating autophagy via inhibiting Akt or Protein Kinase C.  (This is why Blagonosky in NY likes rapamycin over TORC1-specific mTOR inhibitors).

drugsautophagy

Image source  mTOR and autophagy, showing impacts of lithium and rapamycin

b. Metformin – .Autophagy may explain as much as 50% of the benefits (mechanism of action) of Metformin.

Metformin activates AMPK and therefore stimulates autophagy via TORC1-dependent and TORC-1 independent methods (see above).  For this reason, metformin is a good “autophagy drug”.  Metformin probably has many other mechanisms of action, however, which cannot be explained by the induction of autophagy.

signalization-pathways-of-metformin

Image source

c. Resveratrol – Resveratrol directly or indirectly activates the NAD+-dependent deacetylase, SIRT1.

SIRT1 activates autophagy by several different mechanisms, the 4 major ones being deacetylation of multiple cytoplasmic proteins including several involved with autophagy, such as ATG5, ATG7, and ATG8/LC3.  SIRT1 also deacetylates the FOXO transcription factors (FOXO3a, FOXO, and FOXO4), but the FOXO proteins are not required for autophagy induction.  It is likely that the effects of SIRT1 on FOXO deacetylation mediate other beneficial effects of resveratrol (not autophagy).

d. Spermidine – The benefits of spermidine can be partially explained by its effects on autophagy.  Spermidine is a histone acetylase inhibitor.  By inhibiting histone acetylase, spermidine allows for the up regulation of autophagy (Atg) genes.  It appears that like resveratrol, spermidine also stimulates overlapping deacetylation reactions of cytoplasmic proteins. See the 2009 publication Autophagy mediates pharmacological lifespan extension by spermidine and resveratrol.

resveratrolspermidineautophagy

Image source

Microsoft Word - Figure 1

Spermidine and autophagy in normal and diabetic states  Image source

 

e. Lithium – The beneficial effects of Lithium for aging and for bipolar illness may be mediated in part by autophagy(ref).

9.  Exercise can both activate and inhibit autophagy.  

For this reason, the benefits of exercise are mostly due to non-autophagy factors.

Decreased autophagy mechanisms with exercise:  Exercise up regulates mTOR, especially resistance exercises like weight lifting.  Exercise also activates the IGF-1 pathway by increasing growth hormone secretion by the pituitary gland, which then in turn stimulates  IGF-1 production by the liver.  IGF-1 inhibits autophagy via the Insulin/IGF-1/PI3K/Akt pathway.

Increased autophagy mechanisms with exercise:   ROS increases with exercise.  Since ROS activates autophagy, this is one mechanism by  which exercise could activate autophagy, but it is unclear if this activates “selective mitochondrial destruction” this way (i.e. mitophagy).

Hypoxia also activates autophagy via a HIF-1a pathway.  This would occur with exercise if you reached your anaerobic threshold during exercise or did IHT exercise (intermittent hypoxia with exercise).

Conclusion:  Exercise can both inhibit and activate autophagy.  This may be why it is difficult to show exactly how exercise prolongs lifespan.

10.  Autophagy exercises anti-aging effects on postmitotic cells.

– There are primarily 5 cytoprotective effects:

  1. Reduced accumulation of toxic protein aggregates, described above
  2. Destroying bad mitochondria via mitophagy, described above
  3. Reduced apoptosis
  4. Reduced necrosis
  5. Improved hormesis

Cells that do not divide are particularly vulnerable to the build-up of protein aggregates seen in neurodegenerative diseases.  Autophagy inducers such as rapamycin, rapalogs, valproate, and lithium have been shown to help in experimental models of Huntington’s disease, tauopathies, Alzheimer’s disease, and Parkinson’s disease.

When mitochondria are defective due to ROS-induced damage, asymmetric fission occurs, allowing for a good mitochondria and a bad mitochondria to “split up”.  The bad mitochondria has a low membrane potential and is tagged by PINK1 and then ubiquinated by Parkin.  At this point, it is recognized by the autophagy system and is destroyed by macroautophagy.

Autophagy also has an anti-apoptotic function in post mitotic cells.   Autophagy helps damaged cells recover and thereby avoid apoptosis.  Autophagy also has an “anti-necrosis” function in post mitotic cells.

Autophagy is also a stress response involving hormesis.  Hormesis is how low (sublethal) doses of cellular stressors result in an up regulation of cellular stress adaptation mechanisms. See the blog entries Multifactorial hormesis II – Powerpoint presentation and Multifactorial Hormesis – the theory and practice of maintaining health and longevityAutophagy has a hormetic dose response curve.  Depending on the strength or duration of the stressor, autophagy or a negative consequence could ensue, as exemplified in this diagram:

hormesis- 2

Image source

11. Anti-aging effects of Autophagy on Proliferating Cells 

– Autophagy has cytoprotective effects and other unique effects in dividing cells:

  1.  Cytoprotective effects – see #10 above
  2. Reduced stem cell attrition
  3. Reduced ROS-induced cellular senescence
  4. Reduced oncogenic transformation
  5. Improved genetic stability
  6. Increased p62 degradation
  7. Anti-cancer effects via increased oncogene-induced senescence and oncogene-induced apoptosis

With aging, there is a decline in bone marrow stem cell function (hematopoeitic stem cells and mesenchymal stem cells) and stem cell number (MSCs only).  Rapamycin restores the self-renewal capability of hematopoietic stem cells (HSCs).  This improves the function of the immune system, of course assuming a lower dose of rapamycin than the immunosuppressive rapamycin dose given for preventing organ transplant rejection.  Rapamycin can also reverse the stem cell loss that occurs in hair follicles and thereby prevent alopecia.  mTOR accelerates cellular senescence by increasing the expression of p16/INK4a, p19/Arf, and p21/Cip1.  These are all markers of cellular senescence and up regulating these tumor suppressors induces cellular senescence.

The tumor suppressor PTEN is just the opposite, however.  Loss of the tumor suppressor PTEN induces a unique type of cellular senescence called “PTEN loss-induced cellular senescence” (PICS).  PICS occurs with mTOR activation and can be reduced by inhibiting MDM2, which leads to an increase in p53 expression.  This would inhibit autophagy. Rapamycin can preclude  permanent (irreversible) cell-cycle arrrest due to inducible p21 expression.  In this aspect, mTOR decreases proliferative potential and mediates stem cell attrition via senescence.  Rapamycin can suppress this.  This effect may be mediated by autophagy or by an autophagy-independent effect of mTOR inhibition.

More importantly, several oncogenes suppress autophagy.  This includes Akt1, PI3K, Bcl-2 family anti-apoptotic proteins.  Most of the proteins that stimulate autophagy also inhibit oncogenesis.  This includes DAPK1, PTEN, TSC1, TSC2, LKB1/STK11, and Beclin-1.  Autophagy can suppress oncogenesis through cell-autonomous effects described below:

  1. Improved quality control of mitochondria (less baseline ROS production)
  2. Enhanced genetic stability
  3. Removal of potentially oncogenic protein p62 via autophagy.
  4. Autophagy up regulation results in oncogene-induced senescence (via Ras)

The diagram below shows the beneficial effects of autophagy on all cell types, specific benefits in proliferating cells, and specific benefits in post-mitotic cells.

 

Kroemer_2

Systemic Anti-Aging Effects of Autophagy   Image source

 12. Autophagy can reduce age-related dysfunction through systemic effects – 

Autophagy also confers several beneficial anti-aging effects that are not due to cytoprotection, or other localized effects within the cell itself.  This includes the following systemic benefits of autophagy:

  1. Defense against infections
  2. Innate immunity
  3. Inhibition of pro-inflammatory signaling
  4. Neuroendocrine effects of autophagy

Autophagy in dying antigen-presenting cells improves the presentation of the antigens to dendritic cells.  In dendritic cells, autophagy improves antigen presentation to T cells.  Autophagy in dying cells is also required for macrophage clearance of these dead/dying cells.   This is how autophagy reduces inflammation.  Autophagy helps keep ATP production going in these dying cells, providing energy for the key step in the lysophosphatidylcholine “find me” signaling as well as the phosphatidylserine “flip flop” that is the “eat me” recognition signal for macrophage ingestion of the dying/dead cells.  By helping macrophages find these cells and recognize that they are ready for macrophage ingestion, these cells do not rupture and spill their intracytoplasmic contents (this is what causes the inflammation with necrosis, where cell membrane rupture occurs).

When autophagy is working hand-in-hand with apoptosis, no inflammation occurs when a cell dies. This is a key beneficial role of autophagy in reducing inflammation.   The decline in autophagy seen in aging may be in part the cause of age-induced type-2 diabetes.  Here the peripheral tissues become insulin resistant.  This may be due to the hepatic suppression of the Atg7 gene, which results in ER stress and insulin resistance.  Induction of autophagy in specific neural populations may be sufficiency to reduce pathological aging.

 

Kroemer_4

More effects of autophagy     Image source

Beyond its cell-autonomous action, autophagy can reduce age-related dysfunctions through systemic effects. Autophagy may contribute to the clearance of intracellular pathogens and the function of antigen-presenting cells (left), reduce inflammation by several mechanisms (middle), or improve the function of neuroendocrine circuits (right).

13.  Autophagy is necessary for maintaining the health of pools of adult stem cells

Frequent readers of this blog know that the writers believe that age-related decline of the health and differentiation capability of adult stem cells and increasing sensescence of those cells may be responsible for many of the effects we associate with aging.  Thus, the positive roles of autophagy in keeping stem cells viable is of great interest to us.

See the comments under 11 above.  Also, the June 2013 review publication Autophagy in stem cells provides “a comprehensive review of the current understanding of the mechanisms and regulation of autophagy in embryonic stem cells, several tissue stem cells (particularly hematopoietic stem cells), as well as a number of cancer stem cells.”  Another such review is the June 2012 e-publication Tightrope act: autophagy in stem cell renewal, differentiation, proliferation, and aging.

stemcellautophagyL

Image Source  “Tightrope act inhibition of mTOR via caloric restriction (CR) or rapamycin induces autophagy. Autophagy clears away damaged proteins and organelles like defective mitochondria, thereby decreasing ROS levels and reducing genomic damage and cellular senescence, thus playing a crucial role in enhancing stem cell longevity. CR may also have a role in maintaining low levels of p16ink4a, a tumor suppressor protein, thus reducing the risk of cancer and promoting proliferation of stem cells. Oncogenesis is countered by loss of PTEN which elicits a p53-dependent prosenescence response to decrease tumorigenesis(ref)”

Only now are studies beginning to emerge that characterize the detailed roles of autophagy in maintaining stem cell health and differentiation viability.  Autophagy in stem cells recapitulates the current state of understanding:  “As a major intracellular degradation and recycling pathway, autophagy is crucial for maintaining cellular homeostasis as well as remodeling during normal development, and dysfunctions in autophagy have been associated with a variety of pathologies including cancer, inflammatory bowel disease and neurodegenerative disease. Stem cells are unique in their ability to self-renew and differentiate into various cells in the body, which are important in development, tissue renewal and a range of disease processes. Therefore, it is predicted that autophagy would be crucial for the quality control mechanisms and maintenance of cellular homeostasis in various stem cells given their relatively long life in the organisms. In contrast to the extensive body of knowledge available for somatic cells, the role of autophagy in the maintenance and function of stem cells is only beginning to be revealed as a result of recent studies. Here we provide a comprehensive review of the current understanding of the mechanisms and regulation of autophagy in embryonic stem cells, several tissue stem cells (particularly hematopoietic stem cells), as well as a number of cancer stem cells. We discuss how recent studies of different knockout mice models have defined the roles of various autophagy genes and related pathways in the regulation of the maintenance, expansion and differentiation of various stem cells. We also highlight the many unanswered questions that will help to drive further research at the intersection of autophagy and stem cell biology in the near future.”

Another very-recent finding related to autophagy and stem cells is reported in the March 31, 2013 paper FIP200 is required for maintenance and differentiation of postnatal neural stem cells.These data reveal that FIP200-mediated autophagy contributes to the maintenance and functions of NSCs through regulation of oxidative state.” FIP200 is “a gene essential for autophagy induction in mammalian cells.”

Exercising control over autophagy may prove useful for efficiently generating induced pluripotent stem cells.  According to the 2012 publication Autophagy in stem cell maintenance and differentiation: “We also discuss a possible role for autophagy during cellular reprogramming and induced pluripotent stem (iPS) cell generation by taking advantage of ATP generation for chromatin remodeling enzyme activity and mitophagy. Finally, the significance of autophagy modulation is discussed in terms of augmenting efficiency of iPS cell generation and differentiation processes.”

A steady stream of research continues to reveal new insights on the roles that autophagy plays in stem cells.  For example, the April 2013 publication FOXO3A directs a protective autophagy program in haematopoietic stem cells reports: “Here we identify autophagy as an essential mechanism protecting HSCs from metabolic stress. We show that mouse HSCs, in contrast to their short-lived myeloid progeny, robustly induce autophagy after ex vivo cytokine withdrawal and in vivo calorie restriction. We demonstrate that FOXO3A is critical to maintain a gene expression program that poises HSCs for rapid induction of autophagy upon starvation. Notably, we find that old HSCs retain an intact FOXO3A-driven pro-autophagy gene program, and that ongoing autophagy is needed to mitigate an energy crisis and allow their survival. Our results demonstrate that autophagy is essential for the life-long maintenance of the HSC compartment and for supporting an old, failing blood system.”

14.  Autophagy is a key step in activating the Nrf2 pathway.  And Nrf2 expression can in turn regulate autophagy.

The importance of the Nrf2 stress-response pathway and its role in generating health has been one of the frequent topics of discussion in this blog.  See specifically the blog entries The pivotal role of Nrf2. Part 1, Part 2, Part 3, and Nrf2 and cancer chemoprevention by phytochemicals.  We know now that autophagy plays a key role in Nrf2 activation, via p62-dependent autophagic degradation of Keap1.  See, for example, the 2012 publication Sestrins Activate Nrf2 by Promoting p62-Dependent Autophagic Degradation of Keap1 and Prevent Oxidative Liver DamageWe also know that, in turn, Nrf2 expression can regulate autophagy.  See for example the March 2013 publication Regulation of Cigarette Smoke (CS)-Induced Autophagy by Nrf2.

15.  Autophagy and aging

We are starting to understand why autophagy stops working well when a person grows old – why autophagy does not work as well as you age.  Among the reasons are:

a. Failure to form autophagosomes – with aging, there appears to be a failure for autophagosomes to form, possibly due to macroautophagy enhancers (glucagon).

b. Failure of fusion – with aging, there appears to be a failure of lysosomes to fuse with autophagosomes.

c. Negative signaling from insulin or insulin receptors – with aging, insulin signaling or insulin receptor signaling activates mTOR in cells.

d. Mitophagy does not work as well in aging.

e. Autophagy decline probably also results in energy (ATP production) decline.

16.  Practical interventions to promote autophagy

There are a number of practical ways to promote autophagy.  Specifically, in partial recap of the above:

  • Fasting activates Autophagy –   caloric restriction affects 5 molecular pathways that activate autophagy
  • Sunlight, Vitamin D and Klotho activate Autophagy – there are three ways through which UV light, Vitamin D, and the Klotho pathway activate autophagy via inhibiting the insulin/IGF-1 pathway
  • Rapamycin activates Autophagy – there are two ways through which mTOR inhibitors activate autophagy –  TORC1 and TORC2 mechanisms
  • Caffeine activates Autophagy – Caffeine can activate autophagy via an mTOR-dependent mechanism
  • Green tea activates Autophagy – ECGC can activate autophagy via an mTOR-dependent mechanism
  • Metformin activates Autophagy – metformin can activate autophagy via AMPK activation – mTOR-dependent and mTOR-independent mechanisms
  • Lithium activates Autophagy –  lithium and other compounds can activate autophagy by inhibiting inositol monophosphate and lower IP3 levels – an mTOR-independent mechanism
  • Resveratrol activates Autophagy – there are four 4 ways through which resveratrol can activate autophagy – via mTOR-dependent and mTOR-independent mechanisms
  • Spermidine activates Autophagy – how spermidine activates autophagy via histone protein deacetylation – mTOR-indepdendent mechanism
  • Hypoxia activates Autophagy –  intermittent hypoxia can increase autophagy via HIF-1a
  • Phytosubstances which activate the Nrf2 pathway can activate Autophagy.  These are many and include soy products and hot chili peppers.

In addition, these lesser-known substances can also activate autophagy:

Amiodarone low dose Cytoplasm – midstream yes Calcium channel blocker =>  TORC1 inhibition.  Also, a mTOR-independent autophagy inducer

  • Fluspirilene low dose Cytoplasm – midstream yes Dopamine antagnoists  => mTOR-dependent autophagy induction
  • Penitrem A low dose Cytoplasm – midstream yes high conductance Ca++activated K+ channel inhibitor => mTOR-dependent autophagy inducer
  • Perihexilenelowdose Cytoplasm- midstream yes 1. TORC1 inhibition
  • Niclosamidelowdose Cytoplasm- midstream yes 1. TORC1 inhibition
  • Trehalose 100 mM Cytoplasm – midstream supplement 1. activates autophagy via an mTOR-independent mechanism
  • Torin-1 low dose Cytoplasm – midstream no 1. mTOR inhibition (much more potent than rapamycin)
  • Trifluoperazine low dose Cytoplasm – midstream  yes Dopamine antagonists => mTOR-dependent autophagy induction

Wrapping it up

Here are some of the main points above covered:

  • Autophagy is like having a Pac man inside each of your cells, chasing down, eating up and recycling dysfunctional organelles, proteins and protein aggregates.  It has three forms: i. chaperone-mediated autophagy, ii. microautophagy and iii. macroautophagy.  The last is the most important one.
  • Autophagy is a stress response and behaves according to the principles of hormesis.
  • Autophagy can retire and eat up old mitochondria which have become electron-leaking engines.
  • Autophagy solves the problem of high baseline levels of reactive oxygen and nitrogen species.
  • Autophagy  does not require proteins to be unfolded for it to work and therefore can perform housekeeping tasks undoable by the other cell-level house cleaning system, the ubiquitin-proteasome system.
  • Autophagy gets rid of the protein aggregates that can make you loose your memory or walk slow as you grow old – those associated with Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, ALS, CTE, and other neurodegenerative conditions.
  • Autophagy keeps adult stem cells healthy and facilitates their capability to differentiate to make normal somatic body cells.
  • Autophagy prevents inflammation – it works hand-in-hand with apoptosis to help the body get rid of dying cells without inducing cell rupture and inflammation.
  • Autophagy prevents cancer – it helps maintain genetic stability, prevents epigenetic gene silencing.  And it helps promote oncogene-induced cellular senescence for cancer prevention.
  • Autophagy saves the lives of cells by preventing unnecessary cellular apoptosis and cell necrosis.
  • Autophagy is involved in Nrf2 activation and to some extent Nrf2 expression negatively regulates autophagy.
  • Autophagy keeps your bone marrow stem cell population alive and functional.
  • Autophagy helps with infections – it helps clear intracellular pathogens such as bacteria and viruses.
  • Autophagy improves the innate immune response.
  • We are starting to understand why autophagy declines with aging.
  • While autophagy declines with aging, it can exercise multiple effects to slow aging down.  It inhibits the major mechanisms of aging such as cellular senescence, protein aggregate build-up, stem cell loss, epigenetic gene silencing, telomere shortening, and oxidative damage to proteins, lipids, and DNA.
  • There are many practical ways to activate Autophagy like consuming green tea and caffeine, and some less-practical ones.

 

 

About James Watson

I am a physician with a keen interest in the molecular biology of aging. I have specific interests in the theories of antagonistic pleiotropy and hormesis as frameworks to understand cellular senescence and mechanisms for coping with cellular stress. The hormetic “stressors” that I am interested in exploiting at low doses include exercise, hypoxia, intermittent caloric restriction, radiation, etc. I also have a very strong interest in the epigenetic theory of aging and pharmacologic/dietary maintenance of histone acetylation and DNA methylation with age. I also am working on pharmacologic methods to destroy senescent cells and to reactivate quiescent endogenous stem cells. In cases where there is a “stem cell exhaustion” in the specific niche, I am very interested in stem cell therapy (Ex: OA)

Source: Autophagy – the housekeeper in every cell that fights aging | AGINGSCIENCES™ – Anti-Aging Firewalls™

Source: A simple, comprehensive plan to prevent or reverse Alzheimer’s Disease and other neurodegenerative diseases – Part 1: The Plan | AGINGSCIENCES™ – Anti-Aging Firewalls™

 

A simple, comprehensive plan to prevent or reverse Alzheimer’s Disease and other neurodegenerative diseases – Part 1: The Plan

By James P Watson, with contributions and editorial assistance by Vince Giuliano

 INTRODUCTION AND OVERALL PRINCIPLES

This is the first of a pair of blog entries concerned with dementias – neurological diseases including Alzheimer’s Disease (AD) and its cousins.  This Part 1 write-up was inspired by a recent small, non-randomized clinical trial done by Dr. Dale Bredesen that showed true “Reversal of Cognitive Decline” in 9 out of 10 patients with documented cognitive decline (Bredesen, 2014).  Not all of these patients had AD, but all had cognitive decline.  Five had AD, two had SCI (subjective cognitive impairment), and two had MCI (mild cognitive impairment).  Although this study was too small to allow any statistical conclusions, it is the most positive report in a series of disappointing reports on the recent failures of Big Pharma’s monoclonal antibodies against amyloid-beta.  Dale Bredesen’s approach was a multifactorial one – utilizing 24 different approaches to halt or reverse cognitive decline.  We explore those 25 interventions here, focusing on the first 19.  They do not depend on drugs.   The focus of this blog entry is “What can be done about dementias now?”

The forthcoming Part 2 blog entry will provides a detailed discussion of some of the key science related to AD and dementias.  This is the “What is science telling us about dementias?” part which gets quite complex.  We review major theories related to AD there including the Hardy Hypothesis related to amloid beta, the GSK3 theory and more detail on the neuroinflammation theory which we introduce in this Part 1 blog entry.  We expect to emphasize the emerging importance APP (Amloid Precursor Protein).  And we will describe some very recent research that appears to establish that a basic cause of AD is the proliferation in aging of vestigal DNA segments in our genomes (known as LINEs which are long interspersed nuclear elements and SINEs which are short interspersed nuclear elements) with encode over and over again for the production of APP and for the failure of its clearance.  This could well finally explain the role of beta amyloid in AD.

We have published a number of earlier blog entries relating to AD and dementias.  For example, you might want to review my August 2014 blog entry The Amyloid Beta face of Alzheimer’s Disease.

About dementias

Dementia only happens to a minority of the population with aging, but is becoming an ever increasing problem with the explosion in longevity occurring world-wide

Cognitive decline is the major “fear” people have of getting old.  Even individuals with the feared “ApoE4 polymorphism” are not “predestined” to develop Alzheimer’s Disease (AD).  The ApoE4 allele is only a “risk factor” for AD, not the cause of AD.

A common error is that most people view “dementia” and “Alzheimer’s disease” as synonyms, but this is incorrect.  Alzheimer’s disease is only responsible for 60% of cases of dementia in the US and even less of the cases in Japan.  In the US,  Vascular Dementia (VaD) is the second-most common cause of dementia (20%), whereas in Japan, the incidence of AD and VaD is almost the same.  In the US, the remaining 20% of dementia cases are due to several other neurodegenerative diseases such as Lewy Body Dementia (LBD), Parkinson’s disease with dementia (PDwithD), Frontotemporal dementia/ALS spectrum disorder (FTD/ALS), and mixed dementia (which is usually a mixture of AD and VaD).

A portrayal of the breakdown follows.

Image source

In the Middle East and China, VaD is more common than AD.  This was true in Japan two decades ago, but now the ratio of AD to VaD is 1:1.  Since AD and VaD are clearly the leading causes of dementia world-wide, we will focus mostly on these two types of dementia.  Also, the risk factors for AD and VaD overlap and there are cases of “mixed dementia” which include features of both diseases.  AD affects 5.4 million Americans and 30 million globally.  By 2050, these numbers will be 13 million (US) and 160 million (world-wide) (Ferri, 2005). Many experts now regard dementia from neurodegenerative diseases as the 3rd leading cause of death after cardiovascular disease and cancer.  Despite millions of dollars being spent annually on research, the exact causes of these dementias are still unknown, but the number of clues to the causes is growing and we will explore some of the main ones in our Part 2 blog entry.

Neuroinflammation is the most universally accepted explanation for AD

What is clear is that the “universal sign” of all neurodegenerative disease is “neuroinflammation”, which under the microscope is manifested as “gliosis” and is seen with AD, VaD, PD, FTD/ALS, and the type of dementia seen after multiple concussions, which is now called “Chronic Traumatic Encephalopathy” (CTE).  Although they all have different “triggers” for each disease, they all have “neuroinflammation” and histologic signs of gliosis.  We return to neuroinflammation several times as a central theme here and in the Part 2 blog entries.

Another “universal feature” is that all of these disease have familial cases with as few as 5% being genetic (AD) and as many as 50% being genetic (FTD).  In these familial cases, there is most often a genetic mutation that is causal in nature (early onset disease) or a single nucleotide polymorphism (SNP) that is not causal in nature, but predisposes the patient to the disease.   With the exception of CTE (where the primary cause is multiple concussions) and PD (where pesticide exposure, family history of PD, and depression combine to produce an odds ratio OR = 12.0), most of the cases of neurodegenerative dementias remain largely sporadic with unknown specific causation.

Environmental risk factors for neurodegenerative diseases are discussed in the 2005 publication Neurodegenerative Diseases: An Overview of Environmental Risk Factors  and in publications in this list.

Despite millions of dollars being spent annually on research, the exact cause of these dementias are still unknown, but the number of clues to the cause is growing.  What is clear is that the “universal sign” of these neurodegenerative diseases is “neuroinflammation”, which under the microscope is manifested as  “gliosis” and is seen with  AD, VaD, PD, FTD/ALS, and the type of dementia seen after multiple concussions, which is now called “Chronic Traumatic Encephalopathy” (CTE).  Although they all have different “triggers” for each disease, they all have “neuroinflammation” and histologic signs of gliosis.  Another “universal feature” is that all of these disease have familial cases with as few as 5% being genetic (AD) and as many as 50% being genetic (FTD).  In these familial cases, there is most often a genetic mutation that is causal in nature (early onset disease) or a single nucleotide polymorphism (SNP) that is not causal in nature, but predisposes the patient to the disease.

With the exception of CTE (where the primary cause is multiple concussions) and PD (where pesticide exposure, family history of PD, and depression combine to produce an odds ratio OR = 12.0), most of the cases of neurodegenerative dementias remain largely sporadic with unknown specific causation.

Failure of Monotherapeutic Approaches to Neurodegeneration – It is time to consider multiple component therapies

The development of drugs to treat neurodegeneration has probably been the biggest failure of the pharmaceutical industry.  Although there are three FDA-approved drugs for AD, none of them produce anything other than a marginal, unsustained effect on symptoms.  Hundreds of clinical trials for AD have failed over the past two decades, most recently being the large Phase III trials of monoclonal antibodies that target amyloid-beta.  As of today, no drugs have been approved for Frontotemporal dementia, Vascular dementia, and Lewy body dementia.  Only one drug has been approved for Amyotrophic lateral sclerosis (ALS).  All of the clinical trials done for these diseases have largely been with monotherapeutic drug approaches.

We know from the field of cardiovascular disease, cancer, and HIV that single drug therapy for these diseases largely fail.  .  It is now clear that cancer is “incurable” with chemotherapy unless multiple drugs are used.  Combination therapies have become the standard for treating these conditions.  The requirement to combine drug therapies appears to pertain as well to diseases that we cannot “cure” but that are are yet treatable:  we can control the disease and prevent premature death from the disease.  This includes cardiovascular disease, HIV, and a few other glaring chronic diseases.  These diseases like dementias involve simultaneous upregulation or downregulation of hundreds or thousands of genes including protein-producing ones, and simultaneous activation or inhibition of a large multiplicity of pathway.  It is a very tall order to find a single molecule that can have the right effects on so very many different upregulated and downregulated molecules and pathways at the same time.  Yet, Big Pharma by tradition and because of patent law likes to look for single molecules that can be patented and that can make a big differences in a key step in a highly specific disease processes.  But most serious aging-related diseases and dementias don’t offer such an opportunity.

The Multi-factorial approach rather than “single target” approaches to Treating Alzheimer’s Disease

For the same reasons, it makes sense that a single drug made by “Big Pharma” could NOT solve the problems with these neurodegenerative diseases.  Here is a list of 25 different interventions that were combined into one effective program that “reversed” AD in 9 of 10 patients treated in a pilot study at UCLA and the Buck Institute.  None of these involve drugs.  I will include in black, the ones that were recommended by Dr. Dale Bredesen in what he calls the “MEND” program, which is an acronym that stands for “Metabolic Enhancement for NeuroDegeneration”.  You can check out his 2014 paper Reversal of cognitive decline: A novel therapeutic program.

SECTION I PRACTICAL INTERVENTIONS

1.  Eat a low glycemic, low inflammatory, low grain diet – Since sugar triggers insulin release and the insulin receptor triggers brain aging, this is easy to understand. For several complex reasons, certain proteins found only in grains (such as wheat germ, wheat gliadins) also triggers inflammation. The foods that have a high glycemic index or have lots of wheat in them include the following:

High glycemic index foods (these are bad) (and pro-inflammatory nonglycemic foods) Low glycemic index foods (these are good) (and anti-inflammatory foods and beverages)
Sweet Fruit – banannas, oranges, grapefuit Fatty fruit – avocadoes, olives, capers
Orange juice, Apple juice, grape juice Unsweetened coconut milk, soymilk, almond milk
Pancakes, waffles, French toast, toast Scrambled eggs, omelettes, boiled eggs, fried eggs
Candy, Pies, Cake, Ice cream, Sherbert Vegetables – Broccoli, Brussel sprouts, Artichokes
Corn bread, Cornflakes, corn oil Olive oil, Coconut oil extract (MCT oil)
Processed cold cereals – Chex, Raisin bran Oatmeal, barley cereal, rye bread, etc.
   Cream of wheat, Fruit loops, etc. Mushrooms, seaweed (Sushi), cheese, butter
Toast, bread, donuts, bagels, croissants tomato soup (add some protein), mushroom soup
Potatoes, potato chips, French fries Cream of broccoli soup, lentils, legumes
Sweetened yogurt, sweetened milk Unsweetened yogurt, Greek yogurt
Cow’s milk, Chocolate milk, hot cocoa Prosage patties, garden burgers, vegelinks
Jam, jelly, honey, maple syrup, pancake syrup Soymeat, tofu, vegameat, Frichick
Peanut butter, Jam, and bread sandwiches Portobello  mushroom sandwiches w/o bread
White rice, brown rice, pita bread, wild rice Indian curries (leave out the potatoes), Thai curry
Wheat thins, Pretzels, wheat snacks Dried kale chips, seaweed snacks, onion snacks
Sugar drinks, sweetened tea, Gatoraid Green tea, white tea (no caffeine), herbal teas

2.   Enhance autophagy – This can be done without fasting all day.  Research has shown that fasting for at least 12 hours per day (evening and night) is sufficient to activate autophagy.  Not eating for at least 3 hours before bedtime also activates autophagy.  Eating the evening meal earlier in the day also helps.  For those who do not want to fast for at least 12 hours, there may be little hope of “cleaning the cobwebs out of the brain”.  Studies have shown that eating too much or eating late at night completely shuts off autophagy.  This is probably the #1 reason why most people have so much “proteotoxicity” in the brain, the pancreas, and other organs.  You can review our blog entry Autophagy – the housekeeper in every cell that fights aging.

There are some natural compounds and some drugs that stimulate autophagy, however. They include the following:

  • mTOR inhibitors – The mTOR pathway is “downstream” from the Insulin/IGF-1 pathway. The mTOR pathway completely “shuts off” autophagy and stimulates protein synthesis. This is the primary “danger” of eating too much meat or protein (i.e. stimulating the mTOR pathway).  Continually inhibiting the mTOR pathway is probably not a good idea either, since it is very important to synthesize proteins.  However, intermittent mTOR pathway inhibition has been shown to be a very effective way of stimulating “cellular housekeeping” in the brain. The best-known drug that inhibits the mTOR pathway ia rapamycin.  Low glucose levels and low amino acid levels in the blood also inhibit mTOR.  It is interesting that at least one big pharma company, Novartis,  is interested in marketing rapamycin as an anti-aging drug(ref).
  • AMPK activators – The AMPK pathway is one of the major pathways that activates autophagy. AMPK is activated by both exercise and fasting. The AMPK pathway is a “cross-talk” pathway between mTOR and the Insulin/IGF-1 pathway.  Activating AMPK inhibits both of these “bad” pathways. (They are only bad in certain contexts of aging and still serve important functions in aging people.  We could not be alive without them.  In the Part 2 blog entry we will talk about how some times IGF is the good guy we don’t want to be without.)  Besides exercise and fasting, AMPK can be stimulated by three hormones, some drugs and many natural compounds. The most potent AMPK activator is muscle contraction (i.e. exercise). The three hormones that stimulate AMPK are thyroid hormone and two hormones secreted from fat: leptin and adiponectin. Next to this, the most potent chemical activators of AMPK are probably AICAR and ZMP. These are synthetic compounds that are the only true “exercise mimetics”.  ZMP is a derivative of AICAR.  AICAR has been shown to increase endurance in rodents by 44% without exercise.  This is amazing.  Combining AICAR with exercise makes the drug even more effective. Unfortunately, AICAR is very expensive ($350-450/gram).  Common drugs that activate AMPK include metformin and aspirin.  Natural compounds that activated AMPK include resveratrol, pterostilbene, curcumin, EGCG,  betulinic acid, Gynostemma Pentaphyllum, Trans-Tiliroside (from rose hips), and 3-phosphoglycerate.  See this list for articles in this blog that deal with autophagy or describe autophagy activators.
  • Sirtuin activators – The 3rd major family of pathways that activates autophagy is for the Sirtuin enzymes (SIRT1-7). Sirtuins are enzymes that remove acetyl groups from proteins. The most important ones it deacetylates for autophagy are 3 proteins that are crucial to the autophagy system of “cellular housekeeping”.  These 3 proteins are Atg5, Atg7, and Atg8. There are many practical reasons why activating Sirtuin-induced autophagy is critical to health.  For instance, SIRT1 activation protects cells in human degenerative discs from death by promoting autophagy.  This is why fasting has been shown to eliminate back pain. The most well-known SIRT1 activator is resveratrol, the active ingredient in red wine.  However, both red wine and white wine have been shown to activate Sirtuin enzymes.  NAD+, NMN, and NR all activate Sirtuin enzymes (all 7 of them), whereas resveratrol only activates SIRT1.   You can see our blog entry NAD+ an emerging framework for health and life extension — Part 1: The NAD World

3.   Reduce stress – psychological stress, depression, worrying, and being obsessive compulsive all increase the risk of Alzheimer’s disease. The most effective ways to reduce “cellular stress” are as follows:

  • Yoga – yoga has been scientifically proven to reduce stress. The mechanism may be multifactorial, but studies suggest that activating stretch receptors in the muscles induces the SIRT3 gene.  The Sirtuin pathway is a major pathway activated by fasting, caloric restriction, red wine, NAD+, NMN, NR, and certain other natural compounds.
  • Meditation – meditation has been scientifically prove to reduce stress. However, 3 minutes of prayer is NOT meditation. Meditation requires 30-60 minutes of time. The MEND program recommends 20 minutes of meditation twice a day (No one prays that long).
  • Tai chi – this ancient Chinese form of exercise has been shown to reduce stress
  • Exercise followed by rest – exercise alone does not reduce stress, but exercise followed by a good night’s rest is very effective at reducing stress
  • Stretching exercises – These have a special beneficial effect on stress, especially back stretching exercises for back pain.

Self-monitoring of daily stress and exercise can be helpful for knowing what your stress levels are and how good a job you are doing at keeping stress at non-harmful levels.  A great many of the upstream conditions that can lead to dementias mentioned here (sedentary life style, improper diet, inadequate sleep, etc) are likely to induce constitutional stress which can be picked up by such monitoring.  A host of new wearable devices can keep track of exercise and its consequences.  See the blog entry Digital health – health and fitness wearables, apps and platforms – implications for assessing health and longevity interventions – Part 1.  Vince has identified two constitutional stress measurements in his blog entry that can be tracked starting with smartwatch heart rate and sleep measurements, MRHR (morning resting heart rate before awakening), and ERHR-MRHR (difference between evening resting heart rate and morning resting heart rate during sleep, a measure of overnight sleep-related constitutional stress recovery),.  These are described in the blog entry Digital health – health and fitness wearables, Part 2: looking for practical stress biomarkersAlso, heart rate variability is another personally trackable constitutional measurement of stress,  See my recent blog entry on heart rate variability, Digital Health Part 3.

4.    Optimize sleep – At least 8 hours of sleep at night is very effective in preventing Alzheimer’s disease.

Daytime sleeping probably is not as effective, but is probably not harmful provided that a person is not too sedentary with daytime sleeping (i.e. short naps).  Adding 0.5 – 3 mg of melatonin and 500 mg of tryptophan is also very helpful in getting a good night’s sleep.  One of the biggest problems with getting a good night’s sleep is sleep apnea, which is actually very common as we get older.

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“A Simplified schematic of the proposed interventions that may have potential to delay AD pathogenesis — The green arrows indicate pathways for improved circadian regulation and sleep quality, ultimately delaying AD pathogenesis. According to this model, chronobiotics (i.e., bright light therapy (BLT); melatonin; exercise; and food restriction) and good sleep hygiene could be used individually—but preferably in combination—to improve circadian regulation and sleep quality, decrease inflammation and Aβ deposition, and thereby delay AD pathogenesis.”  Image and legend source

5.   Exercise – The World Health Organization recommends 150 minutes of exercise per week, but the best scientific evidence suggests that this is NOT enough. The best scientific evidence suggests at least 450 minutes of exercise per week.  That is 60 minutes per day and an extra 20 minutes on one of those days.  If you want to skip Saturday, that means 75 minutes per day (1hr 15 minutes).  The exercise should include the following for preventing Alzheimer’s disease:

  • Swimming, outdoor hiking, calisthenics, aerobic fitness classes, spinning classes, etc.
  • 30-45 minutes of aerobic exercise where the heart rate is 60% of training heart rate.  This can be on a stationary bicycle, an elipical machine, a “hand bicycle”, a stair climber,
  • 1 mile per day of walking outside (the speed is not important)
  • Resistance exercise – this includes weight lifting, machines, stretch bands, push-ups, etc.
  • Stretching – stretching activates stretch receptors which activates the SIRT3 gene, which is key for mitochondrial function and decreasing free radicals in the muscles (which cause pain
  • Listening to relaxing music – classical music listening is a good way to relax.

Watching TV or looking at a computer screen and “surfing on the computer” probably does NOT work to reduce cellular stress.  Here are some of the blog entries we have published relating to exercise.

6.   Brain stimulation – The Mayo Clinic did a study in 487 patients where they participated in a computerized cognitive training program called “Brain Fitness Program” by Posit Science. This computer training required 1 hour of time per day, 5 days per week for 8 weeks (totaling 40 hours). This was called the IMPACT study.  This program increased their auditory processing speed by 131% and improved their memory an equivalent of approximately 10 years!  Here is some information on this inexpensive computer program:

Some of us think that we may keep our brains fit by constantly trying to figure out the mechanisms of aging.

7.  Keep your homocysteine low – High homocysteine levels seem to correlate with inflammation and also with deficiencies in folate cycle intermediates. The MEND program recommendation is to check your homocysteine levels and if it is > 7, then to take methyl-B12, methyltetrahydrofolate, pyridoxal-5-phosphate, and trimethylglycine (if necessary). The dosages are: Methyltetrahydrofolate – 0.8 mg/day and Pyridoxine-5-phosphate –  50 mg/day

8.   Keep your vitamin B12 high – Vitamin B12 is very important in memory and prevention of dementia. Vit B12 deficiency alone can cause dementia. It is easier to prevent than to reverse.  The MEND program recommends taking methyl-B12, not regular B12. They recommend basing the dose of methyl-B12 on serum levels of B12, which they recommend keeping above 500 with 1mg of methylB12/day.

9.  Keep your C-reactive protein low – CRP is a measure of inflammation. This correlates very well with inflammation in the brain (called neuroinflammation).  They recommend keeping the CRP levels below 1.0 and the Albumin/globulin ratio > 1.5.  There are no FDA-approved drugs that lower this which are safe to be used on a chronic basis.  However, there are several natural products that are effective in reducing C-reactive protein (CRP).  They include curcumin (400 mg/day), Fish oil (DHA & EPA), and an anti-inflammatory diet that is low in sugar and wheat products.  The MEND program recommends 700 mg of DHA twice a day (total 1400 mg) and 500 mg of EPA twice a day (total 1,000 mg).  Since most Fish oil capsules are only about 1/3rd omega-3 fatty acids, that means you need to take about 7,000-8,000 mg (i.e. 7-8 one gram capsules) per day of Fish oil.

10.   Keep your fasting insulin low – Most people develop insulin resistance with aging. Unfortunately, this is rarely diagnosed until they have already suffered the consequences of insulin resistance, which include metabolic syndrome, hypertriglyceridemia, hypercholesterolemia, Alzheimer’s disease osteoarthritis, accelerated hearing loss, accelerated visual impairment (including presbyopia, cataracts, and age-related macular degeneration, aka AMD).  Once these things occur, then reducing your fasting insulin no longer is useful – the cells are already dead!  The MEND program recommends keeping your fasting insulin to < 7.0.  The best way to do this is to eat a low glycemic index diet, encourage ketogenesis by 12 hours of fasting per day, exercise, sleep, and in some cases the drug metformin.  We have found that the NAD precursor, NMN is effective in reducing fasting insulin levels.  Other supplements designed to enhance NAD+ may help as well.

11.   Hormone balancing – The MEND program recommends normalizing thyroid hormone levels (free T3, free T4, estrogen, testosterone, progesterone, pregnenolone, and cortisol). For most people, cortisol levels are way too high.  The best way to reduce cortisol is to reduce stress, improve sleep, and also possibly to supplement with NMN or NR.  The rest of the hormones decline with aging and often need replacement. Here are some ways to make this safe:

  • Testosterone replacement therapy – this is risky in older men, due to the risks of accelerated coronary artery narrowing due to neointimal hyperplasia, as well as benign prostatic hypertrophy worsening or by making prostate cancer grow. For this reason, a thorough work-up for prostate cancer must be done before starting testosterone. In addition, testosterone dosing should be based on testosterone levels.
  • Progesterone – This is primarily for women, but also helps men in low doses. Any progesterone replacement therapy should also be based on blood levels of progesterone.
  • Pregnenolone – This helps both men and women for the brain.
  • Estradiol (E2) – This should also be done based on blood levels of E2

12.   Healthy gut bacteria – Most people have very unhealthy gut bacteria due to the use of antibiotics, due to general anesthesia, and due to dietary factors such as a high sugar diet. As a result, the lactobacillus that are good for your health often die.  In addition, the fiber-fermenting bacteria are often absent, thereby eliminating the healthful effects of a high fiber diet.  Probiotics and prebiotics are often helpful in restoring healthy gut bacteria.  You can see Vince’s 2012 blog entry Gut microbiota, probiotics, prebiotics and synbiotics – keys to health and longevity.

13.   Reducing amyloid beta aggregates – One of the hallmarks of Alzheimer’s disease is the accumulation of misfolded, aggregates of a protein called amyloid beta. Fortunately, there are two natural compounds that if taken in large quantities can reduce amyloid-beta plaques in the brain.  They are Ashwagandha and curcumin.  Both of these are effective in reducing amyloid beta plaques.  The MEND program recommend doses of 500 mg for Ashwagandha and 400 mg for curcumin.  Because curcumin is so poorly absorbed, it is better to take a liposomal or nanoparticle form of the curcumin, like Bio-curcumin 95. Curcumin can be taking as a pill, but it may be absorbed much better in curry that has coconut oil, since the coconut oil creates an emultion and micelles which can be absorbed by the lymphatic system and thereby “bypass” the liver and the “first pass effect”.   Ashwagandha is much better absorbed and does not have as much of a problem. It can be taken as a pill, but also can be taken as a tea.   My friend Dr. Vince Giuliano has made a liposomal form of these two compounds together with two complementary anti-inflammatory herbal extracts which he believes get into the blood stream in concentrations that are 8-10 times higher than by pill form.  He has written about these and other phytosubstances a number of times, e.g.(ref) (ref) (ref) (ref) (ref).

14.   Cognitive enhancement – This category was probably added to the MEND program for supplements that could not be categorized elsewhere. They specifically recommend the natural product called Bacopa monniera and Magnesium. Bacopa monnieri is also called “water hyssop”, “herb of grace”, “Indian pennywort” and Withania somnifera.  Bacopa monniera has been shown to reduce amyloid plaque and prevent synaptic decline in mouse models of AD.  One possible mechanism by which Bacopa monnieri works is to enhance LDL receptor-related protein, which is the “amyloid exporter” in the brain.  There are many studies that show a benefit from Bacopa monniera In humans. A meta-analysis of 6 high quality clinical trials of Bacopa monniera showed that 9 out of 17 tests showed improved performance in the domain of “memory free recall”. In a study on Okadaic acid induced memory impaired rats, the effect of standardized extract of Bacopa monnieri and Melatonin on the Nrf2 pathway was investigated.  “OKA caused a significant memory deficit with oxidative stress, neuroinflammation, and neuronal loss which was concomitant with attenuated expression of Nrf2, HO1, and GCLC. Treatment with BM and Melatonin significantly improved memory dysfunction in OKA rats as shown by decreased latency time and path length. The treatments also restored Nrf2, HO1, and GCLC expressions and decreased oxidative stress, neuroinflammation, and neuronal loss. Thus strengthening the endogenous defense through Nrf2 modulation plays a key role in the protective effect of BM and Melatonin in OKA induced memory impairment in rats.” There is a special form of magnesium which is much better incorporated into the cell called Magnesium-L-threonate, aka MgT.  Both can be taken as a capsule.  The dose Bacopa monniera they recommend is 250 mg/day. However, most of the clinical trials recommend dosages of 300-450 mg/day.

15.  Vitamin D3 –Vitamin D3 seems to be quite different than the other vitamins for a variety of reasons. The most important difference is that Vitamin D levels should be checked and individuals need to adjust their dose based on their serum vitamin D3 levels. To prevent AD, the levels of Vitamin D3 need to be > 50 nmol/L.  The strongest evidence for this comes from two recent studies from 2014.  One was a 5 year study in 1,658 elderly patients who started the study with no dementia. During the 5 years, 171 of the 1,658 developed dementia (10% risk over 5 years).  This study looked at “all cause dementia”, of which 90% is Alzheimer’s dementia (AD) and Vascular dementia (VD).  The risk of developing dementia when serum Vitamin D3 levels were > 50 nmol/L was very low.  However, those with Vit D3  levels between 25 and 50 nmol/L had a 1.53 fold higher risk of developing dementia of any type.  Those with levels below 25 nmol/L had a 2.25 nmol higher risk of developing dementia of any type.  The 2nd study reported in 2014 was from Denmark and followed 10,186 individuals in the Danish population for 30 years.  They looked at the risk of specific kinds of dementia and the relationship to Vitamin D3.  For Alzheimer’s disease (AD), the risk of AD type dementia was 1.25-1.29 fold higher in those with serum Vit D3 levels below 25 nmol/L.  For Vascular Dementia (VD), the risk of VD type dementia was 1.22 fold higher in those with serum Vit D3 levels below 25 nmol/L.  In conclusion, low Vitamin D3 levels is one of the largest risk factors for dementia and the easiest to prevent.  Most people do not get their Vitamin D3 levels checked.  Do you know what yours is?

16.   Increasing Nerve Growth Factor (NGF) Hericium erinaceus and ALCAR — Although there are many growth factors that make nerve cells grow, the most important one is probably Nerve Growth Factor (NGF).  NGF is a growth factor made and secreted by astrocytes in the brain and spinal cord.  NGF enhances neuronal stem cell regeneration of the brain.  Exercise is a potent stimulator of NGF secretion. There are several natural compounds that stimulate nerve growth factor secretion.  They include extracts from the mushroom, Hericium erinaceus. Although there are other edible mushrooms that are good for you, of the 4 edible mushrooms that were studied for their effect on NGF secretion, only Hericium erinaceus induced the secretion of NGF from human astrocytes in the Hippocampus of the brain.  Another compound that stimulates the secretion of NGF is Acetyl-L-carnitine, aka ALCAR.  Acetyl-L-carnitine also helps with neuropathic pain.   In rodent models of Alzheimer’s disease, 150 mg/kg/day of ALCAR induced NGF secretion and increased choline acetyltransferase activity, which increasea acetylcholine levels in the hippocampus.

17.   Provide the substrates for synaptic formation uridine, choline, citocolin, DHA, EPA, and herring roe — The ability to form synaptic connections between neurons is a key part of forming memory. Several key molecules are needed to create these synapses and dendritic spines that are not made by the human body (e.g. DHA) or are made in inadequate amounts (e.g. citicoline).   The omega-3 fatty acid called docosahexaenoic acid (DHA) is probably the “rate-limiting substrate” in the formation of presynaptic and postsynaptic proteins.  DHA alone will increase the formation of synapses and increase cognitive performance in humans and experimental animals, but the addition of two other circulating precursors for phosphatidylcholine also enhance memory formation.  These two other precursors are uridine (which gives rise to brain UTP and CTP) and choline (which gives rise to phosphocholine).   Phosphatidylcholine (PC) is the major phosphatide found in human neuronal connections. The other two major synaptic ingredients are uridine and DHA.  Studies have shown that the aministration of choline, uridine, and DHA together have a greater effect than the sum of the individual effects (i.e. they have a synergistic effect on generating synapses and dendritic spines). DHA alone increased the synthesis of hippocampal phospholipids by 8-75%, with the greatest percentage being in the synthesis if PC (phosphatidylcholine).  There are still controversies as to how much DHA a person should take per day.

The MEND program recommends 320 mg of DHA/day, but other experts recommend as much as 2,000 mg/day of DHA.  Herring roe, the eggs from the Herring forage fish, is another good source of n-3 polyunsaturated fatty acids that have a high phospholipid content.  MOPL 30 is a supplement product made by Artic Nutrition that includes a lot of phospholipids and a 3:1 ratio of DHA:EPA.  The MOPL 30 proprietary supplement not only increased neuronal generation, it also decreased plasma triacylglycerol and non-esterified fatty acids as well as increased HDL-cholesterol.  Although fasting glucose did not change, the glucose measurement on OGTT decreased at 10 minutes and 120 minutes into the test.   Instead of taking herring roe, uridine, or choline, the MEND program recommends citocoline (aka CDP-Choline) an intermediate compound in the generation of phosphatidylcholine from choline (i.e. already half made).  It is marketed under many names worldwide, including Ceraxon, Cognizin, NeurAxon, Somazina, Synapsine, etc. Studies have shown that citocoline increases dopamine receptor densities, prevents memory impairment, improve focus and mental energy.  Citocoline may also help treat attention deficit disorder (ADD).  The MEND program recommends a dose of 500 mg of Citocoline twice a day, 320 mg of DHA per day, and 180 mg of EPA per day.

18.   Optimize antioxidants – mixed tocopherols, tocotrienols, Selium, blueberries, NAC, Vit C, a-lipoic acid.  Although the free radical theory of aging has largely been proven to be incorrect as the “cause of aging”, there is no question that the “effect of aging” includes free radical damage to proteins, lipids, and nucleic acids that make up a cell.  To try to mitigate these “downstream effects” of aging, many believe that the judicious use of antioxidants still plays a useful role in treating neurodegeneration.  In this blog we have questioned that viewpoint and have pointed out that “antioxidants” like those mentioned often have powerful epigenetic impacts that better explain their actions(ref)(ref).

19.  Optimize Zn:fCu ratio – Alzheimer’s disease may be caused (in part) by copper toxicity — The fact that Alzheimer’s disease was rare prior to 1900, yet now being very common has led many experts to look for environmental “causes” of AD. One of the leading “suspects” in a long list of environmental risks for AD is inorganic copper, which comes from drinking water and supplement pills. There is clear evidence from human subjects that serum free copper is elevated with AD and that the level of free copper in the serum correlates with cognition and predicts cognition loss.  Animal studies have replicated these findings and have shown that as little as 0.12 ppm of coper in distilled drinking water in cholesterol-fed rabbits greatly enhanced the formation of AD.

A 2nd feature of AD is that those affected also have Zinc deficiency.  A small clinical trial published in 2014 showed that in patients over the age of 70, Zinc supplementation protected against cognitive loss and also reduced serum free copper levels in AD patients.  For these reasons, it is unclear if the efficacy of Zinc therapy is on restoring normal Zn levels or if it is due to reducing Cu levels.

The following Table lists the remaining interventions in Dale Bredesen’s list.  These are fairly clear and we will not expand on them here.

20.  Ensure nocturnal oxygenation Exclude or treat sleep apnea [54]
21.  Optimize mitochondrial function CoQ or ubiquinol, α-lipoic acid, PQQ, NAC, ALCAR, Se, Zn, resveratrol, ascorbate, thiamine [55]
22.  Increase focus Pantothenic acid Acetylcholine synthesis requirement
23.  Increase SirT1 function Resveratrol [32]
24.  Exclude heavy metal toxicity Evaluate Hg, Pb, Cd; chelate if indicated CNS effects of heavy metals
25.  MCT effects Coconut oil or Axona [56]

Neuroinflammation “causes” all of the neurodegeneratove diseases

Although we will save most of our discussion on the science of AD to the coming Part 2 blog entry in this series, we comment here a bit more on the the science behind most of the above interventions – their neuroinflammatory nature.

In all neurodegeneratiave diseases (both familial and sporadic cases), there is evidence of a chronic, low grade brain inflammation that does not go away.  Histologically, this is called “gliosis”, a term that describes what is seen under the microscope. As mentioned above, microglial cells are increased in number and they appear “angry” (i.e. they are activated) likely due to the presence of 1-42.  It is likely that these microglial cells are secreting pro-inflammatory factors which are causing the inflammation, although the picture is actually much more complex.  Vince has written about this in 2011 and before in the blog entries Key roles of glia and microglia in age-related neurodegenerative diseases, New views of Alzheimer’s disease and new approaches to treating it, and Alzheimer’s Disease Update. We surface some additional insights here and in Part 2..

This illustration portrays some of the inflammatory processes that go on when microglia and astrocytes are activated:

Image and legend  source The 2014 publication Inflammasomes in neuroinflammation and changes in brain function: a focused review  “Cytokines hypothesis of neuroinflammation: Implications in comorbidity of systemic illnesses with psychiatric disorders. Pro-inflammatory cytokines can migrate between systemic circulation and brain in both directions which could explain the comorbidity of systemic illnesses with psychiatric disorders. There are three pathways for the transport of pro-inflammatory cytokines from systemic circulation to brain as described by Capuron and Miller (2011): Cellular, Humoral, and Neural. Moreover, PAMPs and DAMPs from trauma, infection, and metabolic waste can prime glial cells to express pro-inflammatory cytokines TNF-α, IL-1β, and IL-6. When expressed, these cytokines activates granulocytes, monocytes/macrophages, Natural Killer, and T cells and together contribute to the pathophysiology of neuroinflammation. Chronic neuroinflammation could result in neurodegeneration and associated psychiatric disorders. These pro-inflammatory cytokines also stimulate production and expression of anti-inflammatory cytokine by glial cells that function as negative feedback to reduce the expression of pro-inflammatory cytokines, subsiding the neuroinflammation. MCP-1, Monocyte chemoattractant protein-1; CP, Choroid plexus; CVO, Circumventricular organ.”

The chronic inflammation viewpoint of Alzheimer’s disease is related to but somewhat different than the Beta Amloid viewpoint, the viewpoint covered in my recent blog entry The Amyloid Beta face of Alzheimer’s Disease.

The situation is described in a 2014 publication by Landry and Liu-Ambrose: “An alternative to the classic amyloid centric view of AD suggests that late-onset AD results from age-related alterations in innate immunity and chronic systemic inflammation (for review see Krstic and Knuesel, 2013).

In the Part 2 blog entry we will go into the neuroinflammation hypothesis in further depth and will explore other theories as to causes of AD and the other neurodegenerative diseases.

So, a basic strategy for preventing or delaying the onset of neurodegenerative diseases is to mount a multifront war on systematic inflammation.  The 25 Bredesen interventions described above are initiatives in that war.

About James Watson

I am a physician with a keen interest in the molecular biology of aging. I have specific interests in the theories of antagonistic pleiotropy and hormesis as frameworks to understand cellular senescence and mechanisms for coping with cellular stress. The hormetic “stressors” that I am interested in exploiting at low doses include exercise, hypoxia, intermittent caloric restriction, radiation, etc. I also have a very strong interest in the epigenetic theory of aging and pharmacologic/dietary maintenance of histone acetylation and DNA methylation with age. I also am working on pharmacologic methods to destroy senescent cells and to reactivate quiescent endogenous stem cells. In cases where there is a “stem cell exhaustion” in the specific niche, I am very interested in stem cell therapy (Ex: OA)

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Light-harvesting chlorophyll pigments enable mammalian mitochondria to capture photonic energy and produce ATP
Chen Xu, Junhua Zhang, Doina M. Mihai, Ilyas Washington

ABSTRACT

Sunlight is the most abundant energy source on this planet. However, the ability to convert sunlight into biological energy in the form of adenosine-5′-triphosphate (ATP) is thought to be limited to chlorophyll-containing chloroplasts in photosynthetic organisms. Here we show that mammalian mitochondria can also capture light and synthesize ATP when mixed with a light-capturing metabolite of chlorophyll. The same metabolite fed to the worm Caenorhabditis elegans leads to increase in ATP synthesis upon light exposure, along with an increase in life span. We further demonstrate the same potential to convert light into energy exists in mammals, as chlorophyll metabolites accumulate in mice, rats and swine when fed a chlorophyll-rich diet. Results suggest chlorophyll type molecules modulate mitochondrial ATP by catalyzing the reduction of coenzyme Q, a slow step in mitochondrial ATP synthesis. We propose that through consumption of plant chlorophyll pigments, animals, too, are able to derive energy directly from sunlight.

INTRODUCTION

Determining how organisms obtain energy from the environment is fundamental to our understanding of life. In nearly all organisms, energy is stored and transported as adenosine-5′-triphosphate (ATP). In animals, the vast majority of ATP is synthesized in the mitochondria through respiration, a catabolic process. However, plants have co-evolved endosymbiotically to produce chloroplasts, which synthesize light-absorbing chlorophyll molecules that can capture light to use as energy for ATP synthesis. Many animals consume this light-absorbing chlorophyll through their diet. Inside the body, chlorophyll is converted into a variety of metabolites (Ferruzzi and Blakeslee, 2007; Ma and Dolphin, 1999) that retain the ability to absorb light in the visible spectrum at wavelengths that can penetrate into animal tissues. We sought to elucidate the consequences of light absorption by these potential dietary metabolites. We show that dietary metabolites of chlorophyll can enter the circulation, are present in tissues, and can be enriched in the mitochondria. When incubated with a light-capturing metabolite of chlorophyll, isolated mammalian mitochondria and animal-derived tissues, have higher concentrations of ATP when exposed to light, compared with animal tissues not mixed with the metabolite. We demonstrate that the same metabolite increases ATP concentrations, and extends the median life span of Caenorhabditis elegans, upon light exposure; supporting the hypothesis that photonic energy capture through dietary-derived metabolites may be an important means of energy regulation in animals. The presented data are consistent with the hypothesis that metabolites of dietary chlorophyll modulate mitochondrial ATP stores by catalyzing the reduction of coenzyme Q. These findings have implications for our understanding of aging, normal cell function and life on earth.

RESULTS

Light-driven ATP synthesis in isolated mammalian mitochondria

To demonstrate that dietary chlorophyll metabolites can modulate ATP levels, we examined the effects of the chlorophyll metabolite pyropheophorbide-a (P-a) on ATP synthesis in isolated mouse liver mitochondria in the presence of red light (λmax = 670 nm), which chlorin-type molecules such as P-a strongly absorb (Aronoff, 1950), and to which biological tissues are relatively transparent. We used P-a because it is an early metabolite of chlorophyll, however, most known metabolites of chlorophyll can be synthesized from P-a by reactions that normally take place in animal cells. Control samples of mitochondria without P-a, and/or kept in the dark were also assayed. In the presence of P-a, mitochondria exposed to red light produce more ATP than mitochondria without P-a (Fig. 1A) or mitochondria kept in the dark (supplementary material Fig. S1A–D). Mitochondrial membrane potential (Fig. 1B) and oxygen consumption (Fig. 1C) increased upon increased light exposure in P-a-treated mitochondria. Light or P-a alone had no effect on any of the above measures of mitochondrial activity (supplementary material Fig. S1E–G). With too much added P-a, ATP concentrations and the rate of oxygen consumption started to return to the levels in mitochondria not incubated with P-a (supplementary material Fig. S1G). Addition of the electron transport inhibitor, sodium azide, reduced the light- and P-a-fueled oxygen consumption by 57% (supplementary material Fig. S1H–I), consistent with oxygen consumption occurring through the electron transport system. Observations were consistent with enhanced ATP production driven by oxidative phosphorylation.

Fig. 1.

Chlorophyll metabolite P-a allows isolated mouse liver mitochondria to capture light to make ATP. (A) ATP synthesis in mouse liver mitochondria incubated with P-a (treated) and exposed to light compared to controls (no P-a). Light exposure started at time zero and ADP was added at 30 seconds. Aliquots were obtained at times shown and relative ATP levels measured using the firefly luciferase assay. Means and standard deviations are shown for each time point. The experiment was run in triplicate with the same batch of mitochondria. *P<0.05 for treated versus control samples. (B) Mitochondrial membrane potential (Δψm) under different treatments as measured by safranin fluorescence. Lower fluorescence equals higher membrane potential. Mitochondria, with or without P-a, were exposed to light for 2 minutes or kept in the dark. Safranin was added at time zero and safranin fluorescence was continuously measured while samples remained under the light. The experiment was run in triplicate with the same batch of mitochondria. Curves shown are the average traces for triplicate runs. (C) Representative oxygraph trace (black line) for mitochondria treated with 4 µM P-a. The light was turned on or off at the times indicated by the arrows. Steeper slope denotes faster oxygen consumption. Dotted lines show slopes when the light was off. When the light was turned on the slope of the black line increased by twofold. That is, oxygen consumption increased when the light was turned on. When the light was turned off, oxygen consumption returned to baseline levels (i.e. the two gray lines have the same slope).

To determine whether P-a associates with mitochondria, we measured P-a fluorescence at 675 nm in the presence of increasing amounts of heart mitochondrial fragments obtained from sheep (Fig. 2A,B). After increasing the concentration of mitochondria, P-a fluorescence increased abruptly, by fivefold, and quickly reached a plateau (Fig. 2B). The abrupt change in fluorescence reflects a change in the environment of P-a, consistent with its change from an aqueous environment to one in which it is presumably associated with a protein. This threshold-sensitive behavior is consistent with zero-order ultrasensitivity, or positively cooperative binding, as described by Goldbeter and Koshland, and suggests a coordinated interaction between the metabolite and mitochondrial fragments (Goldbeter and Koshland, 1981). In contrast, this threshold sensitivity was not observed when increasing amounts of bovine serum albumin (BSA) were added to a solution of P-a; instead, fluorescence steadily increased (supplementary material Fig. S1J).

Fig. 2.

Cooperative binding of P-a to mitochondrial fragments. (A) Fluorescence spectra of P-a before and after addition of sheep heart mitochondrial fragments. Upon addition of mitochondrial fragments, the fluorescence intensity of P-a increased and shifted to a longer wavelength, and the shape of the curve (ratio of the shoulder to main peak) changed. (B) Ultrasensitive steady state response of the P-a–mitochondrial interaction. We measured fluorescence intensity for a 1 µM P-a solution while increasing the concentration of mitochondrial fragments. A Hill coefficient of 36, with a 95% confidence interval from 7 to 65, was obtained by fitting the data to the Hill equation [y = axb/(cb+xb)+offset]. Fit (R2): 0.96.

Catabolic reduction of coenzyme Q10 (CoQ10) is a rate limiting step in respiration (Crane, 2001). The majority of CoQ10 molecules exist in two alternate states of oxidation: ubiquinone, the oxidized form, and ubiquinol, the reduced form. To show that the P-a metabolite could catalyze the photoreduction of mitochondrial CoQ10, we measured the oxidation state of CoQ10 in the above sheep heart mitochondrial fragments in response to exposure to red light. We exposed the mitochondria to light for 10 minutes and measured the percentage of reduced and oxidized CoQ10 by high performance liquid chromatography (HPLC) (Qu et al., 2013). In the freshly isolated mitochondria fragments, nearly all the CoQ10 was oxidized in the form of ubiquinone. However, when we incubated the mitochondria with P-a and exposed the suspension to light, 46% of CoQ10 was reduced (Table 1, entry 1). In comparison, as a positive control, we energized the mitochondria with glutamate/malate and kept the suspension in the dark, yielding a 75% reduction of CoQ10 within 10 minutes (entry 2). In the absence of light, no reduction occurred (entry 3). Upon denaturing the mitochondrial proteins with heat, no reduction occurred (entry 4). Likewise, there was a lack of CoQ10 reduction with CoQ10, P-a and light in the absence of mitochondria (entry 5). These observations are consistent with the fluorescence data in Fig. 2A,B, showing that mitochondrial proteins sequester and organize P-a. In the absence of added P-a, a 2–14% reduction was observed, depending on the mitochondrial preparation used (entry 6). We attribute this ‘background reduction’ to the actions of endogenous chlorophyll metabolites, which we were able to detect by fluorescence spectroscopy (see Distribution of light-absorbing dietary chlorophyll, below).

Table 1. Photoreduction of CoQ10 is an early event in light-stimulated ATP synthesis

Light-driven ATP synthesis in rodent tissue homogenates

To determine whether chlorophyll metabolites and light could influence ATP production in whole tissues, we treated mouse brain homogenates with P-a and exposed them to 670-nm light. The treated brain homogenates synthesized ATP at a 35% faster rate than a control homogenate that was not incubated with P-a [relative ATP synthesis rates (means with standard error and 95% confidence intervals (CI) were: treated, 171.7±8.1 (CI: 154.6–188.7); control, 111.3±9.1 (CI: 92.5–130.0); Fig. 3A]. No linear correlation between the increase in ATP concentrations and the amount of added P-a was observed. Increasing concentrations of P-a elicited the same increase in ATP (supplementary material Fig. S2A,B).

Fig. 3.

Chlorophyll metabolite P-a allows mouse brain tissue homogenates to capture light to make ATP. (A) ATP synthesis in mouse brain homogenate with light exposure. Homogenates were incubated with ADP ± P-a and exposed to light starting at time zero. Aliquots were withdrawn at the times shown. Relative ATP in the aliquots was measured using the firefly luciferase assay. The experiment was run in triplicate with the same batch of homogenized brains. Means and standard errors are shown for each time point. For the control, the standard errors are smaller than the line markings and thus cannot be seen. *P<0.05 for treated versus untreated samples. (B) Overlay of the absorption spectrum of P-a (dotted line) and the wavelengths tested for ATP production in samples treated with P-a and exposed to light for 20 minutes. Peak ATP production correlated with peak P-a absorption. Experiments were done in triplicate. Means and standard errors were calculated, however, standard errors are smaller than the markings and thus cannot be seen.

To demonstrate that photon absorption by P-a was necessary to enhance ATP production, we exposed the P-a-treated brain homogenates to greenish (500 nm) and red (630, 670 and 690 nm) light, all with the same total energy. Wavelengths of light that were more strongly absorbed by P-a produced the largest increase in ATP. For example, the ATP concentration increased by ∼16-fold during exposure to 670 nm light; relative to the same sample kept in the dark, it increased by two-to-fivefold during exposure to 500, 630 and 690-nm light of equal energy (Fig. 3B).

In addition to brain homogenates, P-a also enhanced ATP production in adipose, lens and heart homogenates (supplementary material Fig. S2C–E). Quantification of ATP by both the luciferase assay and high-performance liquid chromatography (HPLC) gave similar results (supplementary material Fig. S2E–F).

Distribution of light-absorbing dietary chlorophyll

Chlorophylls and its metabolites, both chlorins, have signature absorption and admission spectra (Aronoff, 1950). Namely they absorb strongly (ε≈50,000 M−1 cm−1) at ∼665–670 nm and demonstrate intense fluorescence emissions at ∼675 nm, which differentiate chlorins from endogenous molecules in mammals (Aronoff, 1950). To examine whether dietary chlorophyll and/or its metabolites were present in animal tissue after oral consumption, we fed mice a chlorophyll-rich diet. Brain (Fig. 4A) and fat (Fig. 4C) extracts from these mice exhibited red fluorescence at 675 nm when excited with a 410-nm light [brain: treated, 15.4±6.7 (n = 6); control: 4.2±2.6 (n = 6; means ± s.d.); P<0.01]. The excitation spectrum of this 675-nm peak (Fig. 4B) was similar to that of known chlorophyll metabolites with an intact chlorin ring: with maxima at 408, 504, 535, 562 and 607 nm. This red fluorescence diminished, as measured by the area under the 675 nm peak, when animals were given a chlorophyll-free diet for 2 weeks. Red fluorescence could also be seen using fluorescence imaging; fluorescence was stronger in the bodies and brains of animals fed chlorophyll than in animals given a chlorophyll-poor diet [Fig. 4D; mean gray value in the boxed areas with standard deviation and minimum and maximum gray value shown in brackets were: treated brain, 118 (97–138); control brain, 82 (60–100); treated back fat pad, 116 (97–132) and control back fat pad, 35 (25–46)]. The red fluorescence was enriched in the gut and intestines, consistent with dietary chlorophyll being the source of the fluorescence.

Fig. 4.

Dietary chlorophyll results in chlorophyll-metabolite-like fluorescence in tissues. (A) Representative fluorescence spectra of brain extracts following excitation at 410 nm. Relative peak areas for a total of six control animals fed a chlorophyll-poor diet and six treated animals fed a chlorophyll-rich diet. (B) Representative excitation spectrum (emission at 675 nm) of a brain extract from mice fed a chlorophyll-rich diet. (C) Representative fluorescence spectra of abdominal fat extracts from mice fed chlorophyll-poor and rich diets. (D) A 675±10-nm fluorescence image of skinned mice raised on chlorophyll-rich and -poor diets.

To determine whether the red fluorescence was localized to mitochondria, we measured the relative 675-nm fluorescence in whole liver homogenates and mitochondria isolated from these homogenates. As measured by fluorescence intensity, isolated mitochondria contained 2.3-fold as much of the 675-nm fluorescent metabolite(s) per milligram of protein as did the whole liver homogenate. This observation suggests that P-a was concentrated in the mitochondria, consistent with data summarized in Fig. 2A,B, and literature reports (MacDonald et al., 1999; Tang et al., 2006).

Fat and plasma extracts from rats fed chlorophyll-rich diets were further analyzed by HPLC to elucidate the source of the red 675-nm fluorescence. Fig. 5A shows a representative chromatogram with compounds in the eluting solvent that displayed 675-nm fluorescence when excited with 410-nm light. Rat fat extracts and plasma extracts both contained similar chlorophyll-derived metabolites (similar chromatograms not illustrated). Two groups of compounds eluting at 23–30 minutes and 40–46 minutes were detected. Compounds eluting between 23 and 30 minutes had similar retention times to those of the chlorophyll metabolites without the phytyl tail, with at least one carboxylate group, such as P-a. The absorption spectra (the locations of the absorbance maxima and the Soret-to-Qy-band ratios) of this group of compounds were consistent with demetalated chlorophylls (Rabinowitch, 1944), as shown in Fig. 5B. In addition, the spectra of this group of peaks were indicative of coordination to a metal ion. A representative spectrum of such a presumably metalated metabolite is shown in Fig. 5C, showing a red shifted Soret band, a blue shifted Qy-band and a Soret-to-Qy-band ratio of ∼1. The compounds eluting between 40 and 46 minutes had similar retention times to that of the demetalated chlorophyll-a standard (pheophytin-a). In addition, these compounds partitioned with hexanes (polarity index = 0.1) when mixed with hexanes and acetonitrile (polarity index = 5.8). This latter characteristic is consistent with a lack of a carboxylic acid group, or an esterified P-a, such as pheophytin-a. Similar HPLC chromatograms from fat extracts of swine fed chlorophyll rich diets (Mihai et al., 2013) were recorded (supplementary material Fig. S2G), suggesting that uptake and distribution of chlorophyll metabolites were not unique to mice and rats.

Fig. 5.

Light-absorbing metabolites of chlorophyll are present in adipose tissue. (A) HPLC chromatogram of an adipose extract. 2.5 grams of abdominal adipose tissue from a rat fed a chlorophyll-rich diet was extracted with acetone and the acetone concentrate subjected to HPLC. In the chromatogram, only compounds that displayed 675-nm fluorescence, characteristic of chlorophyll and its metabolites possessing a chlorin ring, are shown. Five major peaks are observed along with several minor peaks. For peaks with letters, the corresponding absorption spectra are shown below. (B–D) Absorption spectra of labeled peaks in A (b–d, respectively). Numbers above peaks are peak maxima in nm. Numbers in the center are the ratios of the Soret band, around 400 nm, to the Qy band at around 655 nm. All spectra are consistent with those of metabolites of chlorophyll. Spectrum C has been assigned to a metalated porphyrin.

We quantified total blood pigments from rats that absorbed at 665 nm. Using an extinction coefficient of 52,000 at 665 nm (Lichtenthaler, 1987), which is typical of chlorophyll-a-derived pheophytins, we estimated a plasma concentration of 0.05 µM in two rats fed a chlorophyll-rich diet. The 665-nm peak was absent in animals fed a chlorophyll-poor diet. The amount of measured total metabolite was five- and two-times higher than that reported for the fat soluble vitamins K (Tovar et al., 2006) and D (Halloran and DeLuca, 1979), respectively, in the rat.

Light-driven ATP synthesis in C. elegans

Next, we used C. elegans to evaluate the effects of light-stimulated ATP production in a complex organism. As C. elegans age, there is a drop in cellular ATP (Braeckman et al., 1999; Braeckman et al., 2002). We hypothesized that the worm would live longer if it could offset this decline in ATP by harvesting light energy for ATP synthesis. As our model system, we used firefly luciferase-expressing C. elegans, which upon incubation with luciferin emit a luminescence that is proportional to their ATP pools (Lagido et al., 2009; Lagido et al., 2008; Lagido et al., 2001). Upon incubation with P-a, worms incorporated the metabolite, as measured by fluorescence spectroscopy (supplementary material Fig. S3A). To determine whether there were changes in ATP stores in response to light, we plated two groups of worms into 96-well plates containing luciferin substrate. We measured worm luminescence at time zero. We then exposed one group to 660-nm light and kept the other in the dark and periodically measured luminescence in both groups of worms (summarized in Fig. 6A,B). To determine whether ATP increased in light-exposed animals, we subtracted the luminescence signal of the worms kept in the dark from that of the worms exposed to light (Fig. 6C). Worms that were given P-a had a statistically significant increase in ATP when exposed to light, whereas control worms showed no increase. The metabolite alone had no effect on ATP levels when the worms were kept in the dark (i.e. luminescence intensity remained constant throughout the experiment). The elevated luminescence signal persisted for 1 hour after the light was turned off, at which time measurement ceased. However, the luminescence intensity did not further increase during the time the light was off. It was unclear whether this persistent signal reflected the kinetics of the luciferase–luciferin reaction, luciferase expression, or actual ATP pools. Thus ATP was quantified by additional methods.

Fig. 6.

P-a treatment enables worms to capture light to generate ATP. Black lines show results from worms incubated with P-a at the indicated concentrations; gray lines show results from worms not incubated with P-a. (A) In vivo, real-time ATP levels in 1-day-old worms were tracked during exposure to light. Luciferase-expressing worms were incubated with luciferin and exposed to light at time zero. Luminescence was measured at the times shown. Data represent triplicate experiments of 12 separate sets of worms plated in 12 wells of a 96-well pate. Means and standard deviations are shown for each of the three separate runs. (B) In vivo, real-time ATP levels in worms kept in the dark. The same experiment as in A in the same 96-well plate, but the worms were kept in the dark. (C) Percentage ATP increase for worms in A relative to worms in B. (D) In vivo, real-time ATP monitoring. Groups of worms were incubated with or without P-a; light exposure began at time zero and in vivo ATP levels were determined at the times shown in each group of worms by measuring worm luminescence after the addition of luciferin. Each time point represents a different group of worms exposed to light for the times shown. Each experiment was performed in triplicate sets of 12; averages and standard deviations are shown. P-values of Student’s t-tests are also shown, representing the significance compared with the controls at the same light exposure. (E) The same experiment as described in D, but using 10-day-old worms.

As an alternative means of determining whether light stimulated ATP synthesis, we plated luciferase-expressing worms into a 96-well plate without the luciferin substrate, and exposed them to light. ATP status was determined at time zero, immediately before light exposure, and at 15-minute intervals for a total of 45 minutes by adding the luciferin substrate to a group of worms and measuring luminescence (Fig. 6D,E). We found an increase in ATP when 5-day-old and 10-day-old adult worms were fed the metabolite and exposed to light.

We further confirmed the in vivo increase in ATP using two additional ex vivo methods. After light treatment, we lysed the worms, extracted their ATP and quantified ATP in the homogenate using either the firefly luciferase assay or HPLC (supplementary material Fig. S3B,C). Both methods were consistent with the in vivo ATP measurements.

In addition to an increase in ATP, worms treated with P-a exhibited a 13% increase in respiration when exposed to light, as measured by oxygen consumption. However, light had no effect on the respiration rates in untreated worms (supplementary material Fig. S3D). This observation is consistent with an increase in ATP through oxidative phosphorylation, in accordance with the mitochondrial data. Despite the increase in ATP, the levels of reactive oxygen species (ROS) were equivalent in treated and untreated worms during 5 hous of light exposure, as measured using 2′,7′-dichlorofluorescin diacetate (supplementary material Fig. S3E). In fact, although the difference was not statistically significant, treated worms exhibited, on average, lower levels of ROS.

Light harvesting to extend life span

We next tested whether photonic energy absorption by P-a could prolong life. Life span measurements were taken in liquid cultures according to the method of Gandhi et al. and Mitchell et al. (Gandhi et al., 1980; Mitchell et al., 1979). Adult worms were incubated with P-a for 24 hours. Beginning at day 5 of adulthood, we exposed the worms to red light in a daily 5 hours:19 hours light∶dark cycle. Control worms were not given P-a or exposed to light, but otherwise were kept under identical conditions. Counts were made at 2- to 3-day intervals and deaths were assumed to have occurred at the midpoint of the interval. To obtain the half-life, we plotted the fraction alive at each count verses time and fitted the data to a two-parameter logistic function, known to accurately fit survival of 95% of the population (Vanfleteren et al., 1998). The group treated with P-a and light had a 17% longer median life span than the group that was not treated with P-a, but exposed to light (Fig. 7A,B). P-a treatment alone, in the absence of light, had no effect on life span (supplementary material Fig. S4B). Light treatment alone decreased life span by 10% (supplementary material Fig. S4B), in accordance with reports that nematodes survive better in complete darkness (Thomas, 1965). This decrease in median life span brought on by light was reversed when the worms were treated with P-a. The increased median life span with light and P-a was reproducible with different batches of worms (supplementary material Fig. S4B–E). Increasing the amount of P-a past a certain threshold, however, lead to a gradual decrease in lifespan approaching that of animals not treated with P-a (supplementary material Fig. S4B,C).

Fig. 7.

P-a and light increase C. elegans median life span. (A) Median life spans of worms treated with P-a and exposed to light versus those exposed to light but not treated with P-a. Numbers in parentheses are 95% confidence intervals (CI). (B) Life span plots of the values used for A. P-value is from an f-test. Experiments were run in triplicate. The L4 molt was used as time zero for life span analysis. Worms were grown in liquid culture at 500 worms/ml. For counting, aliquots were withdrawn and placed in a 96-well plate to give ∼10 worms per well; the worms were scored dead or alive on the basis of their movement, determined with the aid of a light microscope. A total of 60–100 worms, representing 1–2% of the total population, were withdrawn and counted at each time point for each flask.

We also examined life span longitudinally. We placed 6-day-old adult P-a- and non-P-a-treated worms into a 96-well plate, exposed them to red light for 5 hours per day and compared the percentage dead and alive after 15 days. Result: 47% of the P-a-treated worms were alive (175 alive; 200 dead) after 15 days, versus 41% of the control worms (111 alive; 163 dead), consistent with the cross-sectional experiments above.

DISCUSSION

Photoreduction of coenzyme Q

Upon incubation of: (1) isolated mouse mitochondria; (2) mouse brain, heart and lens homogenates; (3) homogenized duck fat; and (4) live C. elegans, with a representative metabolite of chlorophyll, light exposure was able to increased ATP concentrations. These observations in a variety of animal tissues perhaps demonstrate the generality of this phenomenon. To synthesize ATP, mitochondrial NADH reductase (complex I) and succinate reductase (complex II) extract electrons from NADH and succinate, respectively. These electrons are used to reduce mitochondrial CoQ10, resulting in ubiquinol (the reduced form of CoQ10). Ubiquinol shuttles the electrons to cytochrome c reductase (complex III), which uses the electrons to reduce cytochrome c, which shuttles the electrons to cytochrome c oxidase (complex IV), which ultimately donates the electrons to molecular oxygen. As a result of this electron flow, protons are pumped from the mitochondrial matrix into the inner membrane space, generating a trans-membrane potential used to drive the enzyme ATP-synthase.

The ‘pool equation’ of Kröger and Klingenberg describes the total rate of electron transfer: Vobs = VoxVred/(Vox+Vred), where Vred is ubiquinone reduction and Vox is ubiquinol oxidation (Kröger and Klingenberg, 1973). Based on this equation, the major roles of complexes I and II can be considered to maintain the mitochondrial ubiquinol pool, and to reduce ubiquinone, which should result in increased ATP synthesis. We reasoned the reduction of CoQ10 could be a potential step in the respiratory pathway in which chlorophyll metabolites could influence ATP levels, as it is known that chlorophyll-type molecules can photoreduce quinones (Chesnokov et al., 2002; Okayama et al., 1967). Indeed, a primary step during photosynthesis is the reduction of the quinone, plastoquinone, by a photochemically excited chlorophyll a (Witt et al., 1963). We hypothesized that if the reduction of mitochondrial ubiquinone could be catalyzed by a photoactivated chlorophyll metabolite, such as P-a, then ATP synthesis would be driven by light in mitochondria with these dietary metabolites. In the proposed mechanism, electrons would be transferred by a metabolite of chlorophyll to CoQ10, from a chemical oxidant present in the mitochondrial milieu. Many molecules, such as dienols, sulfhydryl compounds, ferrous compounds, NADH, NADPH and ascorbic acid, could all potentially act as electron donors. Throughout mammalian evolution, photons of red light from sunlight have been present deep inside almost every tissue in the body. Photosensitized electron transfer from excited chlorophyll-type molecules is widely hypothesized to be a primitive form of light-to-energy conversion that evolved into photosynthesis (Krasnovsky, 1976). Thus it is tempting to speculate that mammals possess conserved mechanisms to harness photonic energy.

Photoexcitation of chlorophyll and derivatives produces the excited singlet state (*1). Oxidative quenching of this singlet state by ubiquinone is possible. Electron transfer could take place through proteins or in solution. Escape from the charge transfer complex and protonation would yield ubisemiquinone, which accounts for 2–3% of the total ubiquinone content of mitochondria (De Jong and Albracht, 1994). Ubisemiquinone can be reduced to ubiquinol by repeating the above sequence or by disproportionation to give one molecule of ubiquinol and one molecule of ubiquinone. Back-electron transfer, from the photoreduced metabolite to the oxidized quinone, could be inhibited by disproportionation or by organizing the chlorophyll derivative and ubiquinol through protein binding. In line with the CoQ10 photoreduction hypothesis, we observed mitochondrial CoQ10 was reduced when isolated mitochondria were exposed to light and P-a (Table 1). Also consistent with light and/or P-a acting upstream of complexes I and II, in isolated mitochondria we observed an increase in ATP in the absence of added electron transport substrates, such as glutamate and malate (Fig. 1A; supplementary material Fig. S1A–C). However, further evidence is needed to confirm this mechanistic hypothesis.

The effect of light in vivo

Intense red light between 600 and 700 nm has been reported to modulate biological processes (Hashmi et al., 2010; Passarella et al., 1984; Wong-Riley et al., 2005), and has been investigated as a clinical intervention to treat a variety of conditions (Hashmi et al., 2010). Exposure to red light is thought to stimulate cellular energy metabolism and/or energy production by, as yet, poorly defined mechanisms (Hashmi et al., 2010). In the presence of P-a, we observed changes in energetics in animal-derived tissues initiated with light of intensity and wavelengths (≈670 nm at ≈0.8±0.2 W/m2) that can be found in vivo when outdoors on a clear day. On a clear day the amount of light illuminating your brain would allow you to comfortably read a printed book (Benaron et al., 1997). In humans, the temporal bone of the skull and the scalp attenuate only 50% of light at a wavelength of ∼670 nm (Eichler et al., 1977; Wan et al., 1981). In small animals, light can readily reach the entire brain under normal illumination (Berry and Harman, 1956; Massopust and Daigle, 1961; Menaker et al., 1970; Vanbrunt et al., 1964). Sun or room light over the range of 600–700 nm can penetrate an approximately 4-cm-thick abdominal wall with only three-to-five orders of magnitude attenuation (Bearden et al., 2001; Wan et al., 1981). Photons between 630 and 800 nm can penetrate 25 cm through tissue and muscle of the calf (Chance et al., 1988). Adipose tissue is bathed in wavelengths of light that would excite chlorophyll metabolites (Bachem and Reed, 1931; Barun et al., 2007; Zourabian et al., 2000). Thus, identification of pathways, which might have developed to take advantage of this photonic energy, may have far-reaching implications.

Dietary chlorophyll in animals

A potential pathway for photonic energy capture is absorption by dietary-derived plant pigments. Little is known about the pharmacokinetics and pharmacodynamics of dietary chlorophyll or its chlorin-type metabolites in human tissues. Here, we observed the accumulation of chlorin-type molecules in mice, rats and swine administered a diet rich in plant chlorophylls (Figs 4, 5; supplementary material Fig. S2G). Data suggests that sequestration from the diets of chlorophyll-derived molecules, which are capable of absorbing ambient photonic energy, might be a general phenomenon.

To date, the reported chlorophyll metabolites isolated from animals have been demetalated (Egner et al., 2000; Fernandes et al., 2007; Scheie and Flaoyen, 2003). The acidic environment of the stomach is thought to bring about loss of magnesium from the chlorophyll (Ferruzzi and Blakeslee, 2007; Ma and Dolphin, 1999). Our absorbance data from extracted pigments from rat fat is consistent with the presence of chlorophyll metabolites bonded to a metal (Fig. 5). If true, the presence of a metal derivative in fat tissue suggests that the pigment was actively re-metalated to take part in coordination chemistry. The identification of several metabolites in the fat and plasma of rats and swine fed a chlorophyll-rich diet that are similar to ones found in plants is significant. However, the structures of the metabolites remain to be elucidated. Chlorin-type molecules are similar in structure and photophysical properties and thus can carry out similar photochemistry (Gradyushko et al., 1970). Our data demonstrate that dietary metabolites of chlorophyll can be distributed throughout the body where photon absorption may lead to an increase in ATP as demonstrated for the chlorin P-a. Indeed, P-a could have been transformed into other metabolites, as most known metabolites of chlorophyll can be formed from P-a by reactions that normally take place in animal cells.

There relationship between the increase in ATP and the amount of added P-a was not linear (supplementary material Fig. S2A,B). ATP stimulation by light in the presence of P-a better fitted a binary on/off, rather than a graded response to P-a. Increasing concentrations of P-a elicited the same increase in ATP, after light exposure. However, with too much added P-a, ATP levels began to fall. This on/off response was also consistent with the observed cooperative binding mode of P-a with mitochondria fragments, suggesting that the threshold response may be regulated by mitochondrial binding of P-a. If chlorophyll metabolites are found to be involved in energy homeostasis, a better understanding of their pharmacodynamics and pharmacokinetics will be needed.

ATP stores and life span

Light of 670 nm wavelength that penetrates the human body, yields ∼43 kcal/mol (1.18×10−22 kcal/photon). Given estimated concentrations of chlorophyll derivatives in the body (Egner et al., 2000; Fernandes et al., 2007; Scheie and Flaoyen, 2003) and the photon flux at 670 nm (Bachem and Reed, 1931; Barun et al., 2007; Bearden et al., 2001; Benaron et al., 1997; Chance et al., 1988; Eichler et al., 1977; Menaker et al., 1970; Vanbrunt et al., 1964; Wan et al., 1981; Zourabian et al., 2000), each chlorophyll metabolite would be expected to absorb only a few photons per second. As such, one might anticipate negligible amounts of additional energy. Organization of chlorophyll metabolites into supramolecular structures, similar to chlorophyll antenna systems in photosynthetic organisms, would increase the effective cross-sectional area of photon absorption and, thus, photon catch. Indeed, our observed positively cooperative binding with mitochondrial fragments is evidence for such organization. Even so, to approach the rate of ATP synthesis powered by NADH or FADH2, sufficient P-a pigment would have to be added to turn animals green. Nevertheless, in model systems, we measure an increase in ATP upon light absorption and changes in fundamental biology (extention in life span). Regardless of the mechanism by which ATP is increased or the measured amount of the increase, perhaps the larger question is: how much of an increase in ATP is enough to make a biological difference?

In animals, treatment with P-a and light both increased ATP and median life span, suggesting that light in the presence of these light absorbing dietary metabolites can significantly affect fundamental biological processes. We previously observed that chlorophyll metabolites enabled photonic energy capture to enhance vision using a mouse model (Isayama et al., 2006; Washington et al., 2004; Washington et al., 2007). Because ATP can regulate a broad range of biological processes, we suspect that ATP modulation also played a role in vision enhancement. The increase in life span may seem contradictory, given that there are studies suggesting that limiting metabolism and ATP synthesis increases the life span of C. elegans. It has been proposed that the life span of this worm might be determined by the metabolic status during development (Dillin et al., 2002) and that there might be a coupling of a slow early metabolism and longevity (Lee et al., 2003). Other observations have led to the hypothesis that increased life span may be achieved by decreasing total energy expenditure across the worm’s entire life span (Van Raamsdonk et al., 2010). However, most studies decrease ATP synthesis from hatching through genetic engineering. By contrast, here, we were able to increase ATP during adulthood at a time when ATP stores reportedly begin to decline. For example, by day 4 of adulthood, the level of ATP and oxygen consumption can drop by as much as 50% compared to day zero (Braeckman et al., 1999; Braeckman et al., 2002). This difference in timing might account for why we observed an increase in life span in response to an increase in ATP. We note that besides caloric restriction, there are only a few interventions that are known (Petrascheck et al., 2007) to increase life span when given to an adult animal.

Alternative mechanisms of life-span extension cannot be ruled out. For example, an increase in reactive oxygen species (ROS) is thought to increase life span in C. elegans (Heidler et al., 2010; Schulz et al., 2007). Upon photon absorption, metabolites of chlorophyll can transfer energy to oxygen, resulting in the generation of singlet oxygen, a ROS. Thus life-span extension seen here might be a result of an increase in ROS due to the generation of singlet oxygen. However, our published data with blood plasma (Qu et al., 2013) and data here from C. elegans do not show an increase in ROS. As ubiquinol is a potent lipid antioxidant (Frei et al., 1990) any ROS increase might be offset by an increase in ubiquinol, generated from the photoreduction of coenzyme Q. Indeed, by producing ubiquinol, P-a might have increased life span by an alternative method by protecting against long-term oxidative damage, which is also a mechanism that has been shown to increase C. elegans life span (Ishii et al., 2004). Further research will be needed to distinguish between the above possible mechanisms.

Conclusion

Both increased sun exposure (Dhar and Lambert, 2013; John et al., 2004; Kent et al., 2013a; Kent et al., 2013b; Levandovski et al., 2013) and the consumption of green vegetables (Block et al., 1992; Ferruzzi and Blakeslee, 2007; van’t Veer et al., 2000) are correlated with better overall health outcomes in a variety of diseases of aging. These benefits are commonly attributed to an increase in vitamin D from sunlight exposure and consumption of antioxidants from green vegetables. Our work suggests these explanations might be incomplete. Sunlight is the most abundant energy source on this planet. Throughout mammalian evolution, the internal organs of most animals, including humans, have been bathed in photonic energy from the sun. Do animals have metabolic pathways that enable them to take greater advantage of this abundant energy source? The demonstration that: (1) light-sensitive chlorophyll-type molecules are sequestered into animal tissues; (2) in the presence of the chlorophyll metabolite P-a, there is an increase in ATP in isolated animal mitochondria, tissue homogenates and in C. elegans, upon exposure to light of wavelengths absorbed by P-a; and (3) in the presence of P-a, light alters fundamental biology resulting in up to a 17% extension of life span in C. elegans suggests that, similarly to plants and photosynthetic organisms, animals also possess metabolic pathways to derive energy directly from sunlight. Additional studies should confirm these conclusions.

MATERIALS AND METHODS

General procedures

Two light sources were used for all experiments, either a 300 W halogen lamp equipped with a variable transformer and band pass interference filters [500, 632, 670, 690 nm with full-width half maximum (FWHM) of 10 nm] or a 1.70 W, 660 nm, LED light bulb. Luminous power density was set to 0.8±0.2 W/m2 as measured by a LI-250A light meter (LI-COR Biosciences, Lincoln, NE). The intensity of red light used was 30–60 times less than the level of red light that we measured on a clear March afternoon in New York City and is less than the level that several organs are exposed to in vivo. Pyropheophorbide-a (P-a, 95% purity) was obtained from Frontier Scientific, Logan, UT. For all experiments, prior to exposing samples to light, we minimized light exposure by preparing samples/experiments with laboratory lights turned off, using a minimum amount of indirect sunlight that shone through laboratory windows (>0.001 W/m2).

Animal protocols were approved by the Institutional Animal Care and Use Committee of Columbia University. Mice (ICR, Charles River, Wilmington, MA) weighing 22–28 g and rats (Fisher 344, Harlan Teklad, Indianapolis, IN), weighing 300 g were used. Swine, fed a chlorophyll-rich diet have been described previously (Mihai et al., 2013).

Continuous ATP monitoring in isolated mouse liver mitochondria

Mice were fed a chlorophyll-poor, purified rodent diet supplied by Harlan (Indianapolis, IN) for a minimum of 2 weeks. We isolated mouse liver mitochondria by differential centrifugation according to existing procedures (Frezza et al., 2007) and used only preparations with a minimum respiratory control ratio above 4.0 [state III/II, using glutamate (5 mM final) and malate (2.5 mM final) as measured with an oxygen electrode from Qubit Systems Inc., Kingston, ON, Canada]. Mitochondria at a final concentration of ≈1 mg protein/ml as determined by the Coomassie Plus (Bradford) protein assay (Thermo Fisher Scientific, Rockford, IL) in buffer A (0.250 M mannitol, 0.02 M HEPES, 0.01 M KCl, 0.003 M KH2PO4, 0.0015 M MgAc2·H2O, 0.001 M EGTA, 1 mg/ml fatty acid–poor BSA, pH 7.4) were incubated with P-a for 30 minutes at 0°C. ADP was added (0.5 mM final concentration) and then 250 µl aliquots of this suspension were placed in nine wells of a 96-well plate for exposure to light at room temperature. At various times, 20 µl aliquots were withdrawn, added to 150 µl lysis buffer (10 mM Tris, pH 7.5; 100 mM NaCl; 1 mM EDTA and 1% Triton X-100), and ATP levels were determined with a commercial kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. Controls were treated in the same way, except they were: (1) incubated at 0°C without P-a (shown), (2) not exposed to light and (3) were incubated without P-a and not exposed to light.

Membrane potential measurement

Mitochondrial membrane potential was monitored in buffer A as described by Feldkamp et al. (Feldkamp et al., 2005). Measurements were made in a 3 ml cuvette placed inside a fluorescence spectrometer (Fluorormax-4, HORIBA Jobin Yvon, Horiba Scientific, Kyoto, Japan) with a final reaction volume of 1 ml. For light exposure, we used a fiber optic light guide to capture and direct light from a 660 nm LED light bulb into the spectrometer. The end of the fiber optic cable was positioned 1 cm above the reaction mixture. Prior to these experiments, light power was measured 1 cm from the end of the fiber optic cable.

Oxygen consumption measurement

Mitochondrial oxygen consumption was measured using an oxygen electrode cuvette (OX1LP-1 ml; Qubit Systems Inc., Kingston, ON, Canada) according to the manufacturer’s instructions. Reactions were run with mitochondria at a concentration of ≈1 mg protein/m’ in buffer A. For light exposure, a 660-nm LED was directed at the plastic [poly-(methyl methacrylate)] chamber.

For inhibition of respiration, sodium azide was added at a final concentration of 0.005 M from a stock solution in water. Sodium azide inhibits cytochrome oxidase (complex IV): oxygen consumption during state 3 respiration is progressively inhibited by increasing concentrations of azide (Bogucka and Wojtczak, 1966).

Analysis of zero-order ultrasensitivity

Mitochondria from sheep hearts were prepared as previously described (Smith, 1967) on two separate occasions from 2 and 1 sheep heart(s) using ‘Procedure 1’. We used mitochondrial fragments to allow P-a direct access to the respiratory chain, to minimize potential complications due to variable rates of P-a import. Mitochondrial isolation started within 1 hour of the death of the animal and the hearts were transported to the laboratory in a bath of 0.25 M sucrose, 0.1 M tris(hydroxymethyl)aminomethane (Tris) at pH 7.5, which was surrounded by ice. Mitochondria were isolated and stored in 250 µl aliquots at a concentration of ∼60 mg of protein/ml in 300 mM trehalose, 10 mM HEPES–KOH pH 7.7, 10 mM KCl, 1 mM EGTA, 1 mM EDTA and 0.1% BSA at −80°C (Yamaguchi et al., 2007) until use. The thawed mitochondria exhibited a respiratory control ratio of ∼1, indicating mitochondrial fragmentation.

Analysis of coenzyme Q redox status

We used sheep heart mitochondria because they contain relatively large amounts of CoQ10, which expedited analysis. For evaluation of CoQ10 redox ratios, frozen mitochondria were thawed at 37°C and diluted with 500 µl buffer A to create a mitochondrial stock solution, which was kept on ice until use. For reactions, 50 µl of this stock suspension was added to 500 µl of buffer A, containing 0.5 µg/ml antimycin A from a 25 µg/ml stock solution in ethanol. Antimycin A binds to the Qi site of cytochrome c reductase (complex III), thereby inhibiting the upstream oxidation of any produced ubiquinol. Pa was added (25 µM final concentration) from a 1.3 mg/ml stock solution in DMSO. The suspension was added to a test tube, mixtures purged with argon and the reactions initiated by placing the tube between two LED light bulbs (previously described). We irradiated the samples for 10 minutes at room temperature. For negative controls, we repeated the above sequence changing the following: (1) in the absence of light; (2) in the absence of added P-a; (3) with heat denatured mitochondria; and (4) in the absence of added mitochondria but with added coenzyme Q. For a positive control we added 10 µl of a stock solution of 0.25 M glutamate/0.125 M malate in Tris buffer at pH 7. For mitochondrial denaturing, 200 µl of stock mitochondrial suspension was purged with argon and placed in a bath at 70°C for 5 minutes. For control reactions without added mitochondria, a coenzyme Q stock solution in buffer A was prepared by adding ALL-QTM (DSM Nutritinal products, Switzerland), a water-soluble coenzyme Q solution containing 10% coenzyme Q, modified food starch, sucrose and medium chain triglycerides, to buffer A. For these reactions 50 µl of the water-soluble Coenzyme Q stock or the denatured suspension was used as above in place of the mitochondrial stock solution. All reactions were adjusted to give the same amount of coenzyme Q in the reaction mixture as measured by HPLC.

To quantify relative ubiquinone and ubiquinol concentrations, a 50 µl aliquot was taken from the reaction mixture and was added to 200 µl of 0.4 M perchloric acid and 100 µl isopropyl ether containing 1 mg of butylated hydroxytoluene/ml as an antioxidant. The solution was vortexed for 1 minute, centrifuged for 2 minutes at 15,000 r.p.m. and the organic phase analyzed by HPLC. HPLC conditions have been reported previously (Qu et al., 2009; Qu et al., 2011). Briefly, we used an isocratic elutent consisting of 1% sodium acetate 3% glacial acetic acid, 5% butanol in methanol at 0.6 ml/minute. The HPLC column was 50×2.1 mm, C-18, 2.6 u, 100 Å (Phenomenex, Torrance, CA). A PDA detector set at 290 nm for ubiquinol and 275 nm for ubiquinone was used. We determined relative ubiquinol and ubiquinone concentrations by their online absorption spectra using extinction coefficients of 14,200 M−1 cm−1 at 275 nm in ethanol for ubiquinone and 4640 M−1 cm–1 at 290 nm in ethanol for ubiquinol (Lester et al., 1959).

Analysis of ATP synthesis in mouse brain homogenates

To produce homogenates of mouse brain, the frontal lobe was homogenized using two strokes of a Potter S homogenizer (Sartorius AG, Goettingen, Germany) at 4°C (20 mg of brain to 1 ml buffer A). The homogenate (80 µl) was added to buffer A (920 µl) and treated as described above for liver samples. Reactions were run in triplicate and data obtained between 5 and 50 minutes after lysis. ATP production showed a linear increase during this time, which was fitted to a line, the slope of which is reported as the relative ATP synthesis rate.

Analysis of ATP synthesis in mouse lens and heart homogenates

Lenses from mice were homogenized (KONTES® DUALL® tissue grinder with glass pestle) in ATP assay buffer (0.15 mM sucrose, 0.5 mM EDTA, 5 mM magnesium chloride, 7.5 mM sodium phosphate, 2 mM HEPES) at 50 µl buffer per lens. We added 1 µl of P-a stock (1 mM) and 1 µl of ADP stock (10 mM) to 100 µl lens homogenate. The mixture was exposed to red light (671 nm at 0.8 W/m2) or kept in dark for 20 minutes. ATP concentrations were determined using a luciferase-based ATP quantification kit according to the manufacture’s instructions (Life Technologies, Grand Island, NY).

Heart tissue (20 mg) was homogenized as above in 1 ml ATP assay buffer. 10 µl of P-a (1 mM) and 10 µl of ADP (10 mM) and 940 µl of ATP assay buffer were added into 40 µl tissue homogenate. The mixture was exposed to red light and ATP was determined as described above using a luciferase based ATP kit.

Analysis of ATP concentrations in duck adipose

We removed visceral fat from a duck (Anas platyrhynchos domestica) less then 30 minutes after death by decapitation and homogenized the fat at 4°C (without buffer) in a loose-fitting Potter-Elvehjem homogenizer. We then added P-a (70 µl of a 3.3 mg/ml stock solution) and ADP (800 µl of a 10 mg/ml stock solution). The homogenate was divided into two groups: one group was kept in the dark, while the other was exposed to red light (671 nm at 0.8 W/m2); both dishes were kept at 37°C. 200-µl aliquots were taken from each dish and ATP was measured using the luciferase assay or by HPLC, as described in the literature (Ally and Park, 1992).

Analysis of the effect of light wavelength

The entire brain of a mouse was homogenized with a Dounce homogenizer (20 mg of brain to 1 ml buffer C: 0.15 mM sucrose, 0.5 mM EDTA, 5 mM MgCl2, 7.5 mM Na2HPO4, 2 mM HEPES) at 4°C. We took a 40-µl aliquot of the homogenate and added it to 940 µl buffer C. We added 10 µl P-a (from a 1 mM stock in DMSO) and placed the sample on ice for 1 hour. We then added 10 µl ADP (from a 10 mM stock). Five 100-µl portions of the suspension were added to each well of a 96-well plate and exposed to light for 40 minutes. Then, 20-µl aliquots of the mixture were lysed with 200-µl lysis buffer for 1 hour on ice, and ATP levels were determined as above using a luciferase-based ATP kit.

Analysis of red fluorescence in tissue extracts

The chlorophyll-rich diet (Harlan Teklad, Indianapolis, IN) contained 15% by weight spirulina [a food supplement produced from cyanobacteria (Ciferri, 1983)], which is equivalent to ∼0.15% by weight chlorophyll-a. The control diet was a purified diet devoid of dietary chlorophylls (Harlan Teklad). The swine chlorophyll-rich diet has been described previously (Mihai et al., 2013).

For fluorescence spectroscopy, five pigs each were given these respective diets ad libitum for 2 weeks. Whole brain or 2–7 grams of abdominal fat was homogenized with a hand-held homogenizer (Omni Micro Homogenizer (μH), Omni International, Kennesaw, GA), HPLC grade acetone (40 ml) was added and the sample was vortexed for 1 minute. Insoluble material was precipitated by centrifugation and the acetone evaporated with a rotary evaporator. The samples were resuspended in 3 ml chloroform and measured directly.

For HPLC and UV spectroscopy, we extracted 2.5 grams of fat, as described above, from rats or swine that had been given a chlorophyll-rich diet, to give a clear oil. We then added 10 ml of absolute ethanol, cooled the sample to −20°C for 30 minutes, pelleted the insoluble material by centrifugation, separated and evaporated the ethanol with a rotary evaporator and re-suspended the sample in 500 µl of absolute ethanol. For plasma, we added 4 ml of plasma to 1 ml of saturated NaCl and 10 ml ethyl acetate, vortexed the sample for 1 minute and separated the layers by centrifugation. We removed the ethyl acetate layer, evaporated the ethyl acetate and re-suspended the resulting film in 300 µl of absolute ethanol. The samples were then used for HPLC and UV spectroscopy. A Waters (Milford, MA) HPLC system with a 600 pump, a 2475 fluorescent detector, a 2998 photodiode array (PDA) detector and a C18, 2.6 u, 100 Å, 150×2.10 mm column (Phenomenex, Torrance, CA) was used for HPLC. Excitation was set to 410 nm and emission set to 675 nm. Absorbance between 275 and 700 was recorded. We used a mobile phase of acetonitrile containing 10% isopropyl alcohol and 0.1% formic acid (solvent A) and water containing 0.1% formic acid (solvent B). Compounds were eluted at a flow rate of 0.3 ml/minute with a 50∶50 mixture of A∶B for 5 minutes, which was changed linearly to 100∶0, A∶B over 15 minutes. At 35 minutes, the flow was increased to 0.5 ml/minute.

In vivo imaging

Animals were imaged with a Maestro™ In-Vivo Imaging System (CRi, Hopkinton, MA), as described by Bouchard et al.; the animals were skinned to reduce interference from skin autofluoresence (Bouchard et al., 2007).

General C. elegans maintenance

Worms were a gift from Dr Cristina Lagido (Department of Molecular and Cell Biology, University of Aberdeen Institute of Medical Sciences, Foresterhill, Aberdeen, UK) (Lagido et al., 2009; Lagido et al., 2001). Nematode husbandry has been described previously (Wood, 1988). Briefly, animals were maintained on nematode growth medium (NGM) agar (Nunc) using E. coli strain OP50 as a food source. To obtain synchronous populations, we expanded a mixed population on egg yolk plates (Krause, 1995). Worm eggs were isolated from the population by treatment with 1% NaOCl/0.5 M NaOH solution (Emmons et al., 1979) and transferred to a liquid culture with E. coli strain OP50, carbenicillin (50 µg/ml) and amphotericin B (0.1 µg/ml; complete medium).

Real-time ATP monitoring in C. elegans

We administered the P-a chlorophyll metabolite by adding it to the culture medium for a minimum of 24 hours. To confirm P-a uptake, we washed away the culture medium containing P-a, suspended the worms in fresh medium and determined the fluorescence spectra in the worms. Treated worms had signature chlorophyll-derived fluorescence, whereas control worms that were not given P-a exhibited no such fluorescence, confirming metabolite uptake.

Method A

Worms were grown in liquid culture at a density of 10,000 worms/ml. Twenty-four hours before the experiment, the culture was split into control and treatment groups and varying amounts of a P-a stock solution in DMSO were added to the treated groups. Control worms were given DMSO vehicle. Worms were washed with M9 buffer (IPM Scientific, Eldersburg, MD) to remove food and unabsorbed P-a and resuspended at 3000 worms/ml. 50 µl of worm suspension from each of these groups were plated into a well of a 96-well plate. Each experimental group was plated into a minimum of 12 wells. To assay ATP stores by luminescence, 100 µl of luminescence buffer containing D-luciferin was added to each well, according to the literature (Lagido et al., 2009; Lagido et al., 2001) and luminescence was recorded in a plate reader. The luminescence buffer was a citric phosphate buffer at pH 6.5, 1% DMSO, 0.05% Triton X-100 and D-luciferin (100 µM). After initial ATP measurements, half of the worms from each experimental group were exposed to LED light centered at 660 nm at 1±2 W/m2; the other half was kept in the dark by covering the plate with aluminium foil. ATP (luminescence signal) was recorded periodically. The amount of ATP synthesized was reported as the difference within an experimental group between the luminescence signal of worms kept in the dark and the worms exposed to light. All experimental procedures outside of red light exposure were performed under dim light. The experiment was repeated three times with different populations of worms.

Method B

Worms were plated as above, with each experimental group divided into 12 wells of a 96-well plate. Four identical 96-well plates were made, each containing worms treated with varying concentrations of P-a and control worms. At time zero, 100 µl of luminescence buffer was added to a plate and in vivo ATP was assayed as luminescence. The remaining three plates were exposed to light and ATP assays were performed every 15 minutes for 45 minutes by the addition of 100 µl of luminescence buffer and the recording of luminescence.

In vitro ATP monitoring in C. elegans

One-day-old adult worms in liquid culture were incubated with P-a for 24 hours, washed with M9 buffer and re-suspended in M9 buffer at 50,000 worms/ml. The control group was incubated in DMSO vehicle without P-a. 100 µl of each worm suspension was placed into 18 centrifuge tubes. At time zero, six tubes from each group were placed in liquid nitrogen and the remaining tubes exposed to red light. Then, at 15 and 30 minutes, six tubes from each group were placed into liquid nitrogen. To measure ATP, we removed the centrifuge tubes from the liquid nitrogen and placed them in boiling water for 15 minutes to lyse the worms (Artal-Sanz and Tavernarakis, 2009). The resulting solution was cleared by centrifugation for 5 minutes at 15,000 rpm and ATP in the lysate was measured using the luciferase assay according to the manufacturer’s instructions or by HPCL according to established protocols (Ally and Park, 1992).

Analysis of C. elegans oxygen consumption

Oxygen consumption was measured using a Clark-type oxygen electrode (Qubit Systems Inc.), as described (Anderson and Dusenbery, 1977; Zarse et al., 2007). One-day-old adult worms in liquid culture at a density of ∼10,000 worms/ml were incubated with P-a (25 µM) for 24 hours in complete medium. Animals were washed three times with M9 buffer to remove bacteria and excess P-a and resuspended in M9 buffer at 10,000 worms/ml. One-ml aliquots of this suspension were transferred into the respiration chamber and respiration was measured at 25°C for 10 minutes while being exposed to an LED light centered at 660 nm at 1±2 W/m2. The control group was treated in the same way but not incubated with P-a.

Analysis of ROS formation in C. elegans

ROS formation was quantified as described by Schulz et al. (Schulz et al., 2007). Three-day-old worms were synchronized in liquid culture at a density of 500 worms/ml in complete medium, then divided into control and treatments groups. The treatment group was incubated for 24 hours with 12 µM P-a and the control group in DMSO vehicle. Bacterial food and P-a were removed by three repeated washes with M9 and the worms resuspended to 500 worms/ml M9 buffer. 50 µl of the suspension from each group was added to the wells of a 96-well plate with opaque walls and a transparent bottom. A 100 µM 2′,7′-dichlorofluorescin diacetate (Sigma-Aldrich, St. Louis, MO) solution in M9 buffer was prepared from a 100 mM 2′,7′-dichlorofluorescin diacetate stock solution in DMSO. 50 µl of this solution were pipetted into the suspensions, resulting in a final concentration of 50 µM. Additional controls included worms without 2′,7′-dichlorofluorescin diacetate and wells containing 2′,7′-dichlorofluorescin diacetate without animals; these were prepared in parallel. Five replicates were measured for each experimental and control group. Immediately after addition of 2′,7′-dichlorofluorescin diacetate, the fluorescence was measured in a SpectraMax M5 microplate reader (Molecular Devices, LLC, Sunnyvale, CA) at excitation and emission wavelengths of 502 and 523 nm. The plates were then exposed to red LED light and fluorescence was re-measured at 2.5 and 5 hours under conditions equivalent to those used previously.

Life span analysis

Population studies

Life span measurements were performed according to the method of Gandhi et al. and Mitchell et al. (Gandhi et al., 1980; Mitchell et al., 1979) with some modifications. Eggs were harvested and grown in darkness in a liquid culture at room temperature. To prevent any progeny developing, 5-fluoro-2′-deoxyuridine (FUDR) (Sigma-Aldrich, 120 µM final) was added at 35 hours after egg isolation, during the fourth larval molt. At day 4 of adulthood, the culture was split into control and experimental groups. The experimental group was treated with 12 µM P-a from a stock solution in DMSO. The control group was given the DMSO vehicle alone. The treated and control cultures were then split into two or three. The final density of worms in all reaction flasks was 500 worms/ml; each flask contained 10 ml, therefore a total of 5000 worms. The following day (day 5 of adulthood), worms were exposed to LED light centered at 660 nm at 1±2 W/m2 for 5 hours. Light exposure was repeated every day until the end of the experiment. For counting, aliquots were withdrawn and placed in a 96-well plate to give ∼10 worms per well; the worms were scored dead or alive on the basis of their movement, determined with the aid of a light microscope. A total of 60–100 worms (representing 1–2% of the total population) were withdrawn and counted at each time point for each flask. Counts were made at 2–3-day intervals and deaths were assumed to have occurred at the midpoint of the interval. Any larvae that hatched from eggs produced before the FUDR was added remained small in the presence of FUDR and were not counted. We used the L4 molt as time zero for life span analysis. To obtain the half-life, we plotted the fraction alive at each count verses time and fitted the data to a two-parameter logistic function using the software GraphPad Prism (GraphPad Software, Inc., La Jolla, CA). The two-parameter model is known to fit survival of 95% of the population fairly accurately (Vanfleteren et al., 1998). Because changes in environment, such as temperature, worm density and the amount of food, can influence life span, control measurements were conducted at the same time under identical conditions. The concentration of P-a dropped (∼75%) throughout the life span studies and it was not adjusted (supplementary material Fig. S4F).

Life span measurements in 96-well microtiter plates

Life span was measured as described in the literature (Solis and Petrascheck, 2011), except that P-a was added at day 4 and light treatment commenced at day 5. Scoring (fraction alive) was done once on day 15.

Footnotes

  • Competing interests

    The authors declare no competing interests.

  • Author contributions

    C.X. conducted studies with worms, and ATP measurements in mitochondria and tissue homogenates. J.Z. conducted ATP measurements in mitochondria and tissue homogenates. D.M. conducted metabolite-binding distribution studies. I.W. designed and supervised the study and wrote the manuscript.

  • Funding

    This work was supported by the Department of the Navy, Office of Naval Research [grant number N00014-08-1-0150 to I.W.]; the Nanoscale Science and Engineering Initiative of the National Science Foundation [grant numbers CHE-0117752, CHE-0641532 to I.W.]; and the New York State Office of Science, Technology and Academic Research (NYSTAR).

  • Supplementary material available online at http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.134262/-/DC1

  • Received April 30, 2013.
  • Accepted October 15, 2013.

References

Sunlight is the most abundant energy source on this planet. However, the ability to convert sunlight into biological energy in the form of adenosine-5′-triphosphate (ATP) is thought to be limited to chlorophyll-containing chloroplasts in photosynthetic organisms. Here we show that mammalian mitochondria can also capture light and synthesize ATP when mixed with a light-capturing metabolite of chlorophyll. The same metabolite fed to the worm Caenorhabditis elegans leads to increase in ATP synthesis upon light exposure, along with an increase in life span. We further demonstrate the same potential to convert light into energy exists in mammals, as chlorophyll metabolites accumulate in mice, rats and swine when fed a chlorophyll-rich diet. Results suggest chlorophyll type molecules modulate mitochondrial ATP by catalyzing the reduction of coenzyme Q, a slow step in mitochondrial ATP synthesis. We propose that through consumption of plant chlorophyll pigments, animals, too, are able to derive energy directly from sunlight.

Source: Light-harvesting chlorophyll pigments enable mammalian mitochondria to capture photonic energy and produce ATP | Journal of Cell Science

Buy Infectious Burden: The Cause Of Aging And Age-Related Disease: Read Books Reviews – Amazon.com ✓ FREE DELIVERY possible on eligible purchases

Source: Amazon.com: Infectious Burden: The Cause Of Aging And Age-Related Disease eBook: Michael Lustgarten: Kindle Store

Gelatin is my new favorite superfood! It’s rich in protein, great for your joints, skin, hair, digestive system, immune system, and is accessible and affordable

Gelatin may conjure images in your mind of that slimy, unnaturally green and orange dessert you got in the cafeteria with your school lunches, but dessert made with gelatin is just the tip of the iceberg when it comes to using this potent superfood. Gelatin is a byproduct of meat processing and comes from the bones and connective tissues of animals. It may sound unsavory, but these parts of animals are actually extremely good for you.

There was a time when people consumed nearly all of an animal, while today we simply end up with the processed and packaged meat, or muscle tissue. What we are missing out on are the nutrients found in those discarded parts. When you buy and use pure gelatin in your smoothies, you are adding back in important amino acids that have many benefits for your health.

Gelatin is composed primarily of collagen, the protein that makes up the connective tissues, bones, and skin of animals, like cows and chickens. It includes several amino acids, the building blocks of protein, but does not provide a complete complement of those needed in the human diet. Gelatin is highest in the amino acids glycine and proline.

Gelatin is used in many industries, not just for food. It is used in photography, pharmaceuticals, glues, and cosmetics. In the food industry, gelatin is used for its ability to act as a gelling agent. You will find it in the ingredient list for any gummy candy, marshmallows, but also in margarine, yogurt, cream cheese, and low-fat processed foods. Gelatin is not a vegan food, but there are substitutes if you don’t eat meat. These include agar and carrageen, both of which come from seaweed. Gelatins labeled as kosher are often vegan.

Health Benefits of Gelatin

So why should you add gelatin to your smoothie recipes? There are many reasons. The amino acids that are prevalent in gelatin have great health benefits, and most of us don’t get enough of them in our diets. Here are the top health benefits attributed to gelatin.

Protein: Why consider using unnatural, processed protein powders when you have this natural source available? Gelatin contains between six and twelve grams of protein in every tablespoon. It does not include a complete protein, but it is a significant source of many of the amino acids you need in your diet.

Digestion: Gelatin naturally attracts and binds water to itself, which means that when you add it to your diet, foods that you consume move more easily through your digestive tract.

Detoxification: Glycine, one of the main components of gelatin, is an amino acid that helps the liver function well. You need your liver functioning because it is responsible for eliminating toxins from the bloodstream.

Joint Health: There has been some research that indicates gelatin can help promote healthy joints and bones. Gelatin comes from the joints and bones of animals, so it only makes sense that it would help with ours. The proteins in gelatin may help prevent the degeneration of collagen that occurs naturally with aging. It has even been shown that eating gelatin reduces stiffness and pain in joints afflicted with arthritis.

Skin, Hair, and Nails: Eating gelatin regularly promotes healthy skin, nails, and hair because these tissues are made largely of proteins. Gelatin can make hair and nails stronger and less brittle, and improve the elasticity and appearance of skin. Although there is little evidence, some people claim gelatin also reduces the appearance of cellulite.

Sleep: It’s glycine to the rescue again. This amino acid has been proven to increase the quality of sleep when consumed regularly. Getting more glycine in your diet will also reduce your drowsiness during the day.

Weight Loss: Gelatin may increase the amount of human growth hormone that your body produces. This in turn helps to increase your metabolism so that you burn more calories. Additionally, the high protein content of gelatin means that it keeps you feeling full longer.

Ulcers: Gelatin has even been shown to reduce the size of stomach ulcers. It is believed to act by healing and strengthening the mucous lining of the stomach, which helps to heal ulcers, but also to prevent them from forming in the first place.

FAQs

Are all gelatins the same? No. Some gelatins have additives. Look for the highest quality gelatin that is 100 percent pure. See my recommended brands below under where to buy.

Do vegan substitutes for gelatin have the same nutrient profile? Unfortunately, no, the seaweed substitutes do not have the same superfood components that gelatin does. Carrageen and agar are largely made up of complex carbohydrates and fiber. They may not have the same health properties as gelatin, but they can be used when you need a gelling agent for a recipe.

Is bone broth the same thing as gelatin? If you do any research into the health benefits of gelatin, you will see bone broth come up again and again. In the past when people ate most of the parts of animals out of necessity, the bones went into the soup pot to make a broth. This broth contains the same proteins as gelatin. You can make bone broth and get the same health benefits as you will with gelatin, but it’s a lot more work.

What does gelatin taste like? Gelatin has no flavor. The gelatin that you see in dessert packets at the grocery store have had flavorings and sugar added. Pure gelatin without those additives is flavorless, which means you can add it to anything.

Gelatin in Smoothies

You truly can add gelatin to any smoothie you make because it has no flavor. Consider replacing your processed protein powder with this pure and natural protein source that will also give you so many other health benefits.

You can experiment with the amount of gelatin that you add to your recipes, but one tablespoon per smoothie serving is a good general rule. If you find that the gelatin clumps up in your smoothie, there are a couple of things you can try. Dissolve your gelatin in a small amount of warm water, or whatever liquid is going into your smoothie, and then add it to the blender. Another option is to use hydrolysate gelatin, which dissolves in cold liquids.

Here’s one of my favorite gelatin containing smoothies. For all of my smoothies featuring gelatin, click here.

Blueberry Gelatin Protein Smoothie

  • 1 cup milk (coconut, almond, hemp, or my favorite, raw milk)
  • 1/2 avocado, seed removed
  • 1 cup frozen blueberries
  • 1 tablespoon gelatin
  • 1 tablespoon coconut oil
  • A dash of cinnamon
  • A little honey, maple syrup, or stevia to sweeten

Place all of the ingredients into your high-speed blender and blend for around 30-45 seconds or until nice and smooth. Note, if you don’t have a very powerful blender you might want to blend everything except the coconut oil first, then drizzle it on and blend for another 5-10 seconds to avoid clumping.

I’d love to know if you use gelatin in your smoothies or cooking and what health benefits you have experienced – please divulge in the comments below.

Where to Buy Gelatin

Your local health food or grocery store likely carries gelatin. There are two brands of gelatin I recommend. Bernard Jensen’s 100% Pure Gelatin and Great Lakes Kosher Gelatin. Jensen’s gelatin contains a whopping 15 grams of protein per serving (1 tablespoon).

Sources for this blog post include:

http://www.peta.org/living/vegetarian-living/gelatin-alternatives.aspx
http://www.thehealthyhomeeconomist.com/the-crucial-reason-you-need-more-gelatin-in-your-diet/
http://wellnessmama.com/7419/12-uses-for-gelatin/
http://en.wikipedia.org/wiki/Gelatin
http://thecoconutmama.com/2013/01/gelatin-challenge-update-and-q-a/

Source: Gelatin: The Forgotten Protein Packed Superfood

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roundbreaking trial to see if it is possible to regenerate the brains of dead people, has won approval from health watchdogs.

A biotech company in the US has been granted ethical permission to recruit 20 patients who have been declared clinically dead from a traumatic brain injury, to test whether parts of their central nervous system can be brought back to life.

Scientists will use a combination of therapies, which include injecting the brain with stem cells and a cocktail of peptides, as well as deploying lasers and nerve stimulation techniques which have been shown to bring patients out of comas.

The trial participants will have been certified dead and only kept alive through life support. They will be monitored for several months using brain imaging equipment to look for signs of regeneration, particularly in the upper spinal cord – the lowest region of the brain stem which controls independent breathing and heartbeat.

The team believes that the brain stem cells may be able to erase their history and re-start life again, based on their surrounding tissue – a process seen in the animal kingdom in creatures like salamanders who can regrow entire limbs.

Dr Ira Pastor, the CEO of Bioquark Inc. said: “This represents the first trial of its kind and another step towards the eventual reversal of death in our lifetime.

“We just received approval for our first 20 subjects and we hope to start recruiting patients immediately from this first site – we are working with the hospital now to identify families where there may be a religious or medical barrier to organ donation.

“To undertake such a complex initiative, we are combining biologic regenerative medicine tools with other existing medical devices typically used for stimulation of the central nervous system, in patients with other severe disorders of consciousness.

“We hope to see results within the first two to three months.”

A doctors looks at an MRS scan
The patients will be monitored using MRI scans for several months  Credit: Chronis Jons

The ReAnima Project has just received approach from an Institutional Review Board at the National Institutes of Health in the US and in India, and the team plans to start recruiting patients immediately.

The first stage, named ‘First In Human Neuro-Regeneration & Neuro-Reanimation’ will be a non-randomised, single group ‘proof of concept’ and will take place at Anupam Hospital in Rudrapur, Uttarakhand India.

The peptides will be administered into the spinal cord daily via a pump, with the stem cells given bi-weekly, over the course of a 6 week period.

Dr Pastor added: “It is a long term vision of ours that a full recovery in such patients is a possibility, although that is not the focus of this first study – but it is a bridge to that eventuality.”

Brain stem death is when a person no longer has any brain stem functions, and has permanently lost the potential for consciousness and the capacity to breathe.

A person is confirmed as being dead when their brain stem function is permanently lost.

However, although brain dead humans are technically no longer alive, their bodies can often still circulate blood, digest food, excrete waste, balance hormones, grow, sexually mature, heal wounds, spike a fever, and gestate and deliver a baby.

Recent studies have also suggested that some electrical activity and blood flow continues after brain cell death, just not enough to allow for the whole body to function.

And while human beings lack substantial regenerative capabilities in the central nervous system, many non-human species, such as amphibians and certain fish, can repair, regenerate and remodel substantial portions of their brain and brain stem even after critical life-threatening trauma.

“Through our study, we will gain unique insights into the state of human brain death, which will have important connections to future therapeutic development for other severe disorders of consciousness, such as coma, and the vegetative and minimally conscious states, as well as a range of degenerative CNS conditions, including Alzheimer’s and Parkinson’s disease,” added Dr Sergei Paylian, Founder, President, and Chief Science Officer of Bioquark Inc.

Commenting on the trial, Dr Dean Burnett, a neuroscientist at the Cardiff University’s Centre for Medical Education said: “While there have been numerous demonstrations in recent years that the human brain and nervous system may not be as fixed and irreparable as is typically assumed, the idea that brain death could be easily reversed seems very far-fetched, given our current abilities and understanding of neuroscience.

“Saving individual parts might be helpful but it’s a long way from resurrecting a whole working brain, in a functional, undamaged state.

Source: Dead could be brought ‘back to life’ in groundbreaking project 

A 2010 article published in Oncology Reports states pancreatic cancer is among the most aggressive forms of human cancer, characterized by a very high mortality rate. It represents the fourth leading cause of cancer death in United States, killing 32,000 people annually. With a 5-year survival rate of only 3 percent and a median survival rate of less than six months, pancreatic cancer carries one of the poorest prognoses. The diagnosis of pancreatic cancer is one of the worst things a doctor ever has to tell a patient. The only FDA-approved therapies for it, Gemcitabine and Erlotinib, produce objective responses in less than 10 percent of patients, while causing severe side-effects in the majority. There is a desperate need for new options.

Clinical research to test new treatments is split into phases. Phase I trials are just to make sure the treatment is safe, to see how much you can give before it becomes toxic. Curcumin, the natural yellow pigment in the spice turmeric has passed a number of those. In fact, there was so little toxicity, the dosing was limited only by the number of pills patients were willing to swallow.

Phase II trials are conducted to see if the drug actually has an effect. Curcumin did, in 2 of the 21 patients that were evaluated. One patient had a 73 percent tumor reduction, but the effect was short-lived. One lesion remained small, but a curcumin-resistant tumor clone emerged. The other patient, who had a stable disease for over 18 months, showed slow improvement over a year. In fact, the only time that patient’s cancer markers bumped up was during a brief three-week stint where the curcumin was stopped.

Love This? Never Miss Another Story.

So curcumin does seem to help some patients with pancreatic cancer, and most importantly, there appears to be little downside. No curcumin-related toxic effects were observed in up to doses of eight grams per day. What happens after eight grams? We don’t know because no one was willing to take that many pills. The patients were willing to go on one of the nastiest chemotherapy regimens on the planet, but didn’t want to be inconvenienced with swallowing a lot of capsules.

The only surefire way to beat pancreatic cancer is to prevent it in the first place. In 2010 I profiled a study conducted by the National Institutes of Health, the largest such study in history, which found that dietary fat of animal origin was associated with increased pancreatic cancer risk.

Which animal fat is the worst? The second largest study has since chimed in to help answer that question. Researchers found that poultry was the worst, with 72 percent increased risk of pancreatic cancer for every 50 grams of daily poultry consumption. Fifty grams is just about a quarter of a chicken breast. The reason white meat came out worse than red may be because of the cooked meat carcinogens in chicken, the heterocyclic amines that build up in grilled and baked chicken. These mutagenic chemicals have been associated with doubling pancreatic cancer risk.

Meat has been associated with significantly increased risk, whereas fake meat is associated with significantly less risk. Those who eat plant-based meats like veggie burgers or veggie dogs three or more times a week had less than half the risk of fatal pancreatic cancer. Legumes and dried fruit appear to be similarly protective.

My grandfather died of pancreatic cancer. By the time the first symptom arose, a dull ache in his gut, it was too late. That’s why we need to work on preventing it.

Carcinogens in grilled and baked chicken may increase the risk of pancreatic cancer, while curcumin may help even in advanced stages of the disease.

Source: Turmeric Curcumin and Pancreatic Cancer | Care2 Healthy Living

Imagine being charged with a DUI when it’s been hours since you’ve had a drink, only to later discover that your body brews its own alcohol.

That’s what happened to an upstate New York woman when she blew a blood alcohol level more than four times the legal limit. Just before Christmas in Hamburg, New York, a judge dismissed the charges after being presented with evidence the woman suffers from “auto-brewery syndrome.”

“I had never heard of auto-brewery syndrome before this case,” attorney Joseph Marusak told CNN on the condition his client’s identity remain anonymous. “But I knew something was amiss when the hospital police took the woman to wanted to release her immediately because she wasn’t exhibiting any symptoms.”

“That prompts me to get on the Internet and see if there is any sort of explanation for a weird reading,” adds Marusak. “Up pops auto-brewery syndrome and away we go.”

“I’m in touch with about 30 people who believe they have this same syndrome, about 10 of them are diagnosed with it,” said Panola College Dean of Nursing Barbara Cordell, who has studied the syndrome for years. “They can function at alcohol levels such as 0.30 and 0.40 when the average person would be comatose or dying. Part of the mystery of this syndrome is how they can have these extremely high levels and still be walking around and talking.”

Extremely rare condition

Also known as gut-fermentation syndrome, this rare medical condition can occur when abnormal amounts of gastrointestinal yeast convert common food carbohydrates into ethanol. The process is believed to take place in the small bowel, and is vastly different from the normal gut fermentation in the large bowel that gives our bodies energy.

First described in 1912 as “germ carbohydrate fermentation,” it was studied in the 1930s and ’40s as a contributing factor to vitamin deficiencies and irritable bowel syndrome. Cases involving the yeast Candida albicans and Candida krusei have popped up in Japan, and in 2013 Cordell documented the case of a 61-year-old man who had frequent bouts of unexplained drunkenness for years before being diagnosed with an intestinal overabundance of Saccharomyces cerevisiae, or brewer’s yeast, the same yeast used to make beer.

Flat tire a blessing

It was a beautiful fall afternoon in 2014 when Marusak’s client met her husband at a restaurant for food and drinks. She consumed “four drinks between noon and 6 p.m.” says Marusak, “less than one drink an hour. We hired a local pharmacologist who said that a woman of her size and weight having four drinks in that period of time should be between 0.01 and 0.05 blood alcohol levels.” That would be beneath the legally impaired level of 0.08 BAC in New York state.

And here’s the “crazy thing,” says Marusak. “Her husband drives to meet friends and she is driving home. She gets a flat close to home but doesn’t want to change the tire so keeps on driving. Another driver sees her struggling with the car and calls it in as an accident. So if she hadn’t had that flat tire, she’d not know to this day that she has this condition.”

Because she blew a blood alcohol level of nearly 0.40, police procedure is to take the accused to a hospital, as that level is considered extremely life-threatening.

Instead of allowing his wife to be released as the hospital recommended based on her lack of drunken symptoms, the husband asked for tests to be run. Sure enough, Marusak says, the results showed a blood alcohol level of 0.30, hours and hours after her last drink. That prompted Marusak to do his own sleuthing.

“I hired two physician assistants and a person trained in Breathalyzers to watch her and take blood alcohol levels over a 12-hour period and had it run at the same lab used by the prosecution,” said Marusak. “Without any drinks, her blood level was double the legal limit at 9:15 a.m., triple the limit at 6 p.m. and more than four times the legal limit at 8:30 p.m., which correlates with the same time of day that the police pulled her over.”

Even more strange, says Marusak, is the fact that the woman exhibited no signs of the levels until she reached a blood alcohol level of between 0.30 and 0.40.

“That’s when she started to feel a bit wobbly on her feet.” Marusak explains that by pointing to the world of alcoholism, where the bodies of “functioning alcoholics” adapt to the high levels of booze in their blood.

Even though the Hamburg judge dismissed the case against his client, Marusak says it’s not over yet.

“I’ve heard the DA’s office says they plan to appeal. I’ll know more by the middle of January.”

Assistant Erie County District Attorney Christopher Belling confirmed a review of the judge’s decision is underway but declined to comment further.

In the meantime, Marusak’s client is treating her condition with anti-fungal medications and a yeast-free diet with absolutely no sugar, no alcohol and very low carbs. While that works for some, Cordell says, others relapse or find little relief

Source: Woman charged with DUI has ‘auto-brewery syndrome’ – CNN.com

For decades, scientists believed that excess body fat was mere storage for unused calories. However, research conducted over the past 20 years suggests added fat is more than a little extra cushion—fat cells are actually “toxic factories,” each one producing inflammatory cytokines (chemical messengers of inflammation) throughout the body and causing potentially serious damage to your health. It is this understanding that has led experts to more closely examine the effects of being overweight, even when an individual is considered physically fit.

In 1998, the National Institutes of Health (NIH) published Clinical Guidelines on the Identification, Evaluation and Treatment of Overweight and Obesity in Adults. These guidelines noted being overweight but in good physical health would reduce the risk of premature death— in other words, being physically fit mattered more than body fat percentage.

But in 2015, the International Journal of Epidemiology released the results of a study that suggested the “fat but fit” theory wasn’t true, based on the health data of more than 1.3 million Swedish men whom researchers followed for 30 years. Those study authors found that the beneficial effects of exercise declined as obesity rates increased. Compared to physically fit obese men, normal-weight men who were not physically fit had a lower risk of dying.

These results are backed by a prior study published in January 2015 that identified a link between increased levels of fat in the body— regardless of physical fitness— and high levels of inflammation. Inflammation is the root cause of all disease, especially chronic conditions, such as heart disease, diabetes, cancer and Alzheimer’s disease. Another study published in the journal Clinical Cancer Research in 2015 observed a correlation between increased levels of white fat tissue and poorer prognosis in early-stage breast cancer. White fat, known as white adipose tissue, is fat stored for energy, but it also plays a role in raising inflammation levels when found in excess throughout the body.

Abdominal obesity, which is fat centralized in the belly, is a sign of high levels of visceral fat in the body. Visceral fat is the type of fat that accumulates in arteries and around organs, and has been credited with increased inflammation and disease risk. Emerging research has found that while this still holds true, fat may be further differentiated. A December 2014 study found that fat deposits may exist on the surface of the myocardium (muscular wall of the heart) and be contained completely beneath the membrane that encloses the heart— in contact with major coronary arteries and their branches. This fat, known as epicardial adipose tissue (EAT), is highly correlated with obesity, and thought to play a role in the development and vulnerability of plaque in the coronary arteries.

More on this…

  • Losing weight? 10 ways to do it cheaper

  • Facebook users recruit friends for diet, supplement programs — but is it legit?

  • 7 reasons why you’re working out and still not losing weight

If being fit doesn’t protect against the dangers of excess weight gain, what can?

While fitness is still an important component of optimal health, it is not a standalone marker.

If you are struggling with losing weight, you will reap significant benefits by increasing lean body mass with exercise.

Here are 3 other tactics that can help you lose weight and lower your disease risk:

1. Assess body fat rather than BMI
One of the primary challenges facing the nation today is the standard of measurement for obesity. At present, obesity is defined by body mass index (BMI), which is essentially a height-to-weight ratio. For example, a man who is 5 feet 10 inches tall weighs 220 pounds and has 12 percent body fat would be considered obese, according to the BMI scale. However, anyone with 12 percent body fat is not overweight or obese. This person is likely a bodybuilder with very high levels of lean muscle. His body fat percentage is a better indicator of his health risk. BMI drastically underscores fat levels in the aging population, particularly postmenopausal women who have lost substantial muscle mass that has been replaced with fat and yet their weight remains steady.

A bioelectrial impedance assessment (BIA) is a more comprehensive look at body composition, assessing lean body mass, body fat, and body water percentages, as well as showing where primary fat stores exist. These assessments are generally available through a physician’s office. Monitoring your body fat rather than BMI will help you better assess your overall health and weight management goals.

2. Add a probiotic to your supplement regimen
Research continues to identify the gut flora as a contributing factor to multiple aspects of health, including weight management and inflammation levels. Unfortunately, the typical American diet often leads to imbalances in the microbiota of the gut favoring the development of intestinal inflammation and increased risk of disease. A daily probiotic (not a dairy-based, sugar-laden probiotic) can help promote healthy bacteria in the gut. According to one study, the Lactobacillus plantarum strain offers the greatest potential for suppressing chronic inflammation in the gut. In November 2015, one study uncovered evidence that the landscape of the bacteria in your gut may be the greatest factor in determining which foods will optimally improve an individual’s weight and general health.

3. Consume a clean, nutrient-rich, whole-foods diet
While certain research may say that the Mediterranean diet is good for some people and that the Paleo diet is good for others, one fact remains: Whole foods are best. Strive to consume a wide variety of fresh vegetables and low-sugar fruits organically or locally sourced. Enjoy a mix of lean proteins from animal sources along with plant-based proteins that are high in fiber, like quinoa. Keep sugar, artificial sweeteners and ingredients, and processed foods out of your diet. These foods contribute to toxins in the body and negatively impact healthy gut microbiota.

Achieving optimal health is always a work in progress. Set small goals every month, week, and day that will drive progress. You don’t have to be perfect, but you should try to make everyday choices, a choice that will maximize your wellbeing— mind, body, and spirit.

Dr. Jennifer Landa is Chief Medical Officer of BodyLogicMD, the nation’s largest franchise of physicians specializing in bioidentical hormone therapy. Dr. Jen spent 10 years as a traditional OB-GYN, and then became board-certified in regenerative medicine, with an emphasis on bio-identical hormones, preventative medicine and nutrition. She is the author of “The Sex Drive Solution for Women.”  Learn more about her programs at www.jenlandamd.com.

For decades, scientists believed that excess body fat was mere storage for unused calories.

Source: ‘Fat but fit’: How carrying excess weight can have long-term health consequences | Fox News

Dr. Gerd Lindner and Dr. Beren Ataç

Desmond  just sent me an e-mail with the below summary of an interview that he conducted with Dr. Gerd Lindner (who works with Dr. Roland Lauster) and his PhD candidate student (now doctor?) Beren Ataç at the recent WCHRS2014 in South Korea.  At the end of this post, I have embedded the video of Dr. Ataç’s presentation that was also filmed by Desmond.

FYI — Dr. Ataç’s Phd thesis was titled: “Development of a vascularized human hair follicle equivalent” and her mentors for that project included Dr. Gerd Lindner and Dr. Roland Lauster.

From Desmond:

Here’s my recount of the discussion I had with Dr Linder & Dr Atac about their work.

Firstly, it is with great excitement to mention that their work into regeneration of a hair follicle did not stop in 2010 after their ground breaking paper was published but rather continued at a remarkable pace with significant breakthroughs being made and some patents filed. Their presentation at the congress gave a great insight into how far along they actually are. It is also important to mention that their lab is subdivided into several teams, each working on regenerating a particular organ of the body such as the liver, kidney and of course the hair follicle.

Their aim is to have at least 10 organ models that are of human origin in order to provide a much better prediction of how a drug would perform in a clinical trial compared to animal studies. A FDA study showed that more than 92% of substances tested in animals show false negative results, and have to be excluded from use in/on humans because of toxic effects. They gave a few examples of where investigational drugs showed to be safe in animal studies but proved to be fatal in human subjects. Tegenero trial being an example.

The hair follicle team (Dr Lindner, Lauster & Atac) have FOUR goals:

1) To create a microchip system where many organs thrive.

2) To create a human hair follicle model that allows rapid screening of compounds that may have an impact on hair regeneration or removal! This may be performed on a single follicle or on a follicle embedded in an engineered full thickness skin equivalent

3) To engineer neopapillae (ECM coated dermal papilla cell spheroids) that will be transplantable into human subjects for patients suffering from Androgenetic Alopecia.

4) and ultimately, to have personalised chips of all genetic backgrounds to give a full picture of pharmacokinetics & pharmacodynamics of an investigational drug.

As for what they have achieved so far:

1) In 2010: Their original paper was published which we are well aware of.

2) In 2011: They bioengineered “human micro-hair follicles” in vitro. These micro-follicles displayed key characteristics of human vellus-like hair follicles. Mesenchymal, ectodermal and neuro-ectodermal originated primary cells from dissected human hair follicles were isolated and expanded. Dermal papilla fibroblasts were kept under low adherent culture conditions (along the same line as the EVAL scaffolds of the Taiwanese that we came across) resulting in the formation of dermal papilla-like aggregates. They then forced keratinocytes and melanocytes to attach to these dermal papilla spheres to allow further follicular development. The result was a self-organizing micro-organoid made up of separate segments enclosed by extracellular matrix membranes, sheath formations and a hair shaft–like fiber. Central ECM proteins and defined mesenchymal and epithelial markers were expressed. Furthermore, inner root sheath formation was found to be present and the melanocyte markers “p-Mel17”, “c-kit” and “TRP-1” were expressed in the supra-papillary region of the microfollicle. These results showed that the de novo formation of human microfollicles in vitro is possible and contains all the basic hair follicle like characteristics.

At this point they realised that after the addition of keratinocytes and melanocytes, the self-organizing micro-organoids followed a stringent pattern of follicular-like formation by generating polarized segments, sheath formations and the production of a hair shaft-like fiber. But the bio-engineered hairs were vellus-like and didn’t turn terminal. This is most probably due to lack of nutrient and oxygen supply during cell culture but may also be caused by an altered gene expression, a problem that Dr Jahoda’s team faced a few years later with their 3D hanging drop spheroid cultures.

Since then, they transferred their culturing method to a perfused bioreactor system and finally came to the conclusion that the best way to improve the microfollicle development is by also co-culturing endothelial cells with the hair follicle which turn into micro-blood vessels and are normally feeding the hair follicles the necessary oxygen, hormones and nutrients. In fact, our hair follicles are very well vascularised, and one can see where they are coming from.

3) So in 2013, they went at it again. They again used an ultra-low adherent attachment conditions. The low-adherent surface which is polycarbonate-based mimics mesenchymal condensation during embryonic development. Under these conditions, DP cells self-aggregate and are then coated with keratinocytes, melanocytes and endothelial cells. After 48 hours the newly formed micro-follicles are placed in a multi-organ chip platform to grow. They also used a new 3D matrix environment to enhance gene expression. These micro-follicles were cultured for 14 days, which showed further improvements in hair follicle-like expressions as you’ll see in the presentation.

So, I guess although they haven’t managed to completely replicate a fully functional (terminal) hair follicle, these follicles look very promising indeed. Some may even call it the endgame (of chess), where there are very few pieces left to play. Exciting times indeed and what a wonderful team of individuals working on such a revolutionary project. The Lauster team as we know them is made up of some great minds: Dr Gerd Lindner and Beren Atac to name a few. I wish them all the very best and I’m sure they’ll have very exciting news to share with the world in a few years.

https://youtu.be/BgASnUOUMN4

Source: June | 2014 | The End of Hair Loss and Balding by 2020

Getting too little sleep during the week can increase some risk factors for diabetes, but sleeping late on weekends might help improve the picture, a small U.S. study suggests.

Researchers conducted a sleep experiment with 19 healthy young men and found just four nights of sleep deprivation were linked to changes in their blood suggesting their bodies weren’t handling sugar as well as usual.

But then, when they let the men get extra sleep for the next two nights, their blood tests returned to normal, countering the effect of the short-term sleep deprivation.

“It gives us some hope that if there is no way to extend sleep during the week, people should try very hard to protect their sleep when they do get an opportunity to sleep in and sleep as much as possible to pay back the sleep debt,” said lead study author Josaine Broussard of the University of Colorado Boulder.

The study doesn’t prove sleeping late every weekend can counter the ill effects of insufficient rest every other night of the week, Broussard cautioned.

And it doesn’t prove that catching up on sleep will prevent diabetes.

“We don’t know if people can recover if the behavior is repeated every week,” Broussard added by email. “It is likely though that if any group of people suffer from sleep loss, getting extra sleep will be beneficial.”

To assess the impact of sleep on diabetes risk, Broussard and colleagues focused on what’s known as insulin sensitivity, or the body’s ability to use the hormone insulin to regulate blood sugar. Impaired insulin sensitivity is one risk factor for type 2 diabetes, which is associated with age and obesity and happens when the body can’t properly convert blood sugar into energy.

The researchers did two brief sleep experiments. On one occasion, the volunteers were permitted just 4.5 hours of rest for four nights, followed by two evenings of extended sleep that amounted to 9.7 hours on average. On another occasion, the same men were allowed to sleep 8.5 hours for four nights.

After the four nights of sleep deprivation, the volunteers’ insulin sensitivity had fallen by 23 percent and their bodies had started to produce extra insulin. But when researchers checked again after two nights of extended rest, the men’s insulin sensitivity, and the amount of insulin their bodies produced, had returned to normal, mirroring what was seen during the portion of the experiment when the volunteers consistently got a good nights’ rest.

The volunteers were given a calorie-controlled diet to limit the potential for their food and drink choices to influence the outcomes. In the real world, when people don’t get enough sleep they tend to overeat, which may limit how much results from this lab experiment might happen in reality, the authors note in a report scheduled for publication in the journal Diabetes Care.

“The results from the present study are unlikely to be fully reflective of what may occur in persons who are older, overweight or obese, or have other potent risk factors for diabetes,” said James Gangwisch, a researcher at Columbia University who wasn’t involved in the study.

Chronically sleep-deprived people are more likely to develop other health problems, though, ranging from obesity to high blood pressure to cognitive deficits, the study authors point out.

“By catching up on sleep on the weekends, people are reducing average extent and severity of the effects of sleep deprivation,” Gangwisch added by email. “Ideally, we would all get sufficient sleep on a nightly basis.”

Getting too little sleep during the week can increase some risk factors for diabetes, but sleeping late on weekends might help improve the picture, a small U.S. study suggests.

Source: Sleeping in on weekends may help reduce diabetes risk | Reuters

Early markers of heart disease are worse with depressive symptoms, but that association was lessened or eliminated with regular physical activity, an observational study showed.

Higher Beck Depression Inventory-II scores correlated with more inflammation as indicated by C-reactive protein levels (P<0.001), more oxidative stress assessed by lower antioxidant glutathione (P<0.001), and poorer vascular function measured by both the augmentation index and subendocardial viability ratio (P=0.008 and P=0.001).

Those associations persisted through adjustment for a number of variables, including weight, age, and some cardiovascular risk factors, Arshed A. Quyyumi, MD, of Emory Clinical Cardiovascular Research Institute in Atlanta, and colleagues reported in a research letter in the Jan. 19 issue of the Journal of the American College of Cardiology.

But getting the recommended level of physical activity interacted significantly with depressive symptom scores for inflammation and cardiac function.

“Thus, vascular stiffening and systemic inflammation that accompany worsening depressive symptoms were more pronounced in sedentary subjects, and these relationships were attenuated in subjects engaged in regular moderate to vigorous physical activity,” the researchers wrote.

“Our findings highlight potential mechanisms by which depressive disorders are linked to cardiovascular disease risk, and support the routine assessment of depressive symptoms to improve cardiovascular disease risk stratification,” they concluded. Physical exercise appears to prevent the adverse cardiovascular consequences of depression, but these findings need to be confirmed in a randomized trial.”

Their study included 965 individuals (median age 49) free of heart disease, cerebrovascular, or peripheral arterial disease at baseline who hadn’t previously been diagnosed with any affective, psychotic, or anxiety disorder.

Activity trims inflammation and cardiac function associations

Source: Could Exercise Shut Down Heart Effects of Depression? | Medpage Today

The Project Avalon Community Forum

Project Avalon Community Forum

Source: The Project Avalon Community Forum

Very high doses of vitamin D may help critically ill patients with respiratory failure leave the hospital sooner, a small study suggests.

Vitamin D is thought to increase the ability of immune cells to fight infection—but hospitalized patients often have insufficient levels of it because of their lack of exercise and exposure to the sun.

For the study, 31 patients were divided into three groups. Two of the groups received high doses of vitamin D3 (a total of 250,000 or 500,000 international units over five days), and one received a placebo.

Significantly shorter stay

“These dosages were significantly higher than normal daily doses and were intended to quickly restore vitamin D levels in patients who have low levels,” says Jenny Han, assistant professor of medicine at Emory University.

Study participants who received a placebo had an average blood level of 21 ng/ml vitamin D. The Endocrine Society defines deficiency as less than 20 ng/ml and insufficiency as between 20 and 30 ng/ml.

The average length of hospital stay was statistically significant for those who received the higher dose of vitamin D: 36 days for placebo, 25 days for lower dose, and 18 days for higher dose.

The length of stay in intensive care also tended to decrease (average 23 days for placebo, 18 days for lower dose vitamin D, 15 days for higher dose vitamin D), but the change was not statistically significant. A similar result was observed for duration of ventilator support.

The majority of the patients in the study had severe sepsis or septic shock; 43 percent had some type of infection upon admission. Some had cardiovascular or neurologic diseases.

More research is needed to determine the effect of vitamin D on patient recovery, the authors say.

“These data can inform the design of a larger, adequately powered randomized controlled trial on the efficacy of high-dose vitamin D3 on host immunity and other indices associated with recovery,” they write.

The findings were presented at a recent meeting of the American Thoracic Society.

Can a super-dose of vitamin D cut hospital stays? – Futurity.

Scientists from Sheffield University say their low-intensity ultrasound device can reduce the healing time of skin ulcers and bedsores by as much as 30%, according to a university news release.

The handheld device was developed by Mark Bass, a PhD in biochemistry at the British university, along with several other colleagues. Bass and his team found that ultrasound technologies transmit a vibration through the skin, waking up the cells inside the wound site which can stimulate and accelerate the healing process. The discovery falls in line with several other efforts over the last few years that look to enhance the healing process, including a study that worked with a nanoparticle platform that researchers found could also accelerate the healing process.

However this technology could set itself apart because it involves a handheld device that could be used to treat patients who suffer from painful skin wounds, particularly diabetic and elderly patients. Skin ulcers occur frequently in patients suffering from diabetes, and can not only be painful, but can escalate to the point of requiring amputation—something that could be avoided with this new handy device that could cut a third of the healing time off.

The key to its development actually sprung from the idea of fooling damaged cells into believing they are at a much earlier stage in the body’s life cycle than they actually are. It’s been known for some time that humans possess much higher regenerative capabilities earlier in life. In fact, the human body actually possesses the capability of perfect scar-free healing while still in the womb—a notion Bass and his team kept in mind when trying to manipulate the damaged cells at the site of a wound.

The group found that when they subjected the wounded area to nano-vibrations, they would cause channels to open within the cellular membrane of the surrounding skin cells, allowing calcium to flow across the membrane. This calcium plays a key role in many of the signalling mechanisms within the cell, which in turn endows the cell with a new front-back orientation. This new orientation causes the cells to move toward the damaged site, effectively pulling the edges of the wound together much like sutures would.

Of course, accelerated wound healing isn’t the only progress being made in the realm of regenerative medicine. Just last year scientists at the University of Edinburgh were able to successfully grow a fully functioning organ from transplanted lab-created cells in a living animal. While growing organs in a controlled environment had been done previously, this marked the first occasion that such a feat was accomplished inside a living mammal.

The common denominator is that regenerative medicine and enhanced healing technologies are quickly rising to the fore, as researchers look to push the boundaries of treatment to new heights.

Bass noted that it is possible to enhance the effects of this ultrasound technology even further with continued refinement of the device and how it is used. Most notably, because ultrasound is relatively risk free, he believes we could see such a device in broad clinical use within the next three or four years.

First Functioning Organ Grown in Living Animal

 

Scientists at the University of Edinburgh have successfully grown a fully functioning organ from transplanted laboratory-created cells in a living animal. While researchers have grown organs in controlled lab environments, this marks the first time that an organ has been created within a living mammal.

Researchers created a thymus, an organ located next to the heart that produces important immune cells, known as T cells, which are vital for guarding against disease.

The scientists were able to take cells called fibroblasts, and turn them into thymus cells in lab mice. Thymus cells are completely different kind of cell from fibroblasts, which were created in this experiment using reprogramming. The reprogrammed thymus cells were capable of supporting development of T cells, a specialized function that only thymus cells can perform, according to materials from the university recounted by a press release from the University of Edinburgh.

thymus
On the left, specialized thymus cells were grown after reprogramming fibroblasts. On the right, lab-grown cells were implanted into a mouse kidney to create a functional “mini-thymus” in a living animal. Credit: MRC Centre for Regenerative Medicine, University of Edinburgh.

Once researchers mixed these reprogrammed cells with other key thymus cell types and transplanted them into a mouse, the cells formed a replacement organ. The new organ had the same structure, complexity, and functionality as a healthy adult thymus. Researchers hope that with further study, the discovery may lead to new treatments for those with a weakened immune system.

This is the first time researchers have created an entire living organ from cells that were created outside of the body through the process of reprogramming. The technique may also offer a way of making patient-matched T cells in the laboratory that could be used in patient-specific cell therapies.

This discovery could prove to be groundbreaking in the growing exploration of regenerative medicine. Recently engineers at MIT designed a biodegradable implantable tissue that can naturally grow bone in the body. The potential impact of lab-created organs and tissue could be monumental when considering the growing number of patients around the world various awaiting transplants.

Despite the various challenges to mass-producing lab-created organs, recent developments such as the lab-generated thymus could prove to be a major step in the direction of organ development and replacement. It also has opened the door to generating specific cells in a lab, paving the way for new innovative cell therapies.

All of these efforts are done with the end goal being to harness the body’s own repair mechanisms, and learning how to manipulate and control these mechanisms to treat diseases. Once these lab-created cells are introduced to the body, they can serve as a catalyst for the immune system, sparking T cell creation and other natural therapeutic responses.

Using Ultrasound to Boost Wound Healing | Qmed.

Earlier this month a guy named Todd Fassler was bitten by a rattlesnake in San Diego, KGTV San Diego reports. In itself this isn’t terribly unusual—the CDC estimates that roughly 7,000 to 8,000 people a year get bit by a venomous snake in the U.S. And somewhere between five and six people die from these bites each year.

What raised eyebrows, though, was Fassler’s hospital bill—all $153,000 of it. KGTV reporter Dan Haggerty shared it on Twitter. Take a look.

It’s not clear whether Fassler has insurance—and whether these are dollar amounts that he will in fact have to pay out of pocket. But the confusion over health care pricing is common for Americans who receive bills and can’t be sure where the numbers come from. I reached out to Fassler for comment but he wasn’t immediately available.

Here’s what we do know based on that photo: The bulk of his hospital bill—$83,000 of it— is due to pharmacy charges. Specifically, charges for the antivenin used to treat the bite. KGTV reports that Fassler depleted the antivenin supplies at two local hospitals during his five-day visit. Nobody expects antivenin to be cheap. But $83,000?

There’s currently only one commercially-available antivenin for treating venomous snakebites in the U.S. — CroFab, manufactured by U.K.-based BTG plc. And with a stable market of 7,000 to 8,000 snakebite victims per year and no competitors, business is pretty good. BTG’s latest annual report shows CroFab sales topped out at close to $63 million British pounds, or $98 million dollars last fiscal year. The antivenin costs hospitals roughly $2,300 per vial, according to Bloomberg, with a typical dose requiring four to six vials. In some cases multiple doses are needed, according to CroFab’s promotional website.

BTG has fought aggressively to keep competitors off the market. A competing product, Anavip, just received FDA approval this year and likely won’t be on the market until late 2018. This lack of competition is one reason why snakebite treatments rack up such huge hospital bills — $55,000. $89,000. $143,000. In May of this year, a snakebit Missouri man died after refusing to seek medical care, saying he couldn’t afford the bill.

But the other reason why hospitals charge so much is the byzantine negotiating process that happens between hospitals and insurance companies to determine the final payout amount. In the case of the $143,000 snakebite in 2012, for instance, Scripps Hospital in San Diego explained that “it is important to understand that these charges are not reflective of what Scripps will be paid. At this time, the patient’s insurance company has not yet paid the bill, and Scripps is in negotiations with the company for the final amount.”

In many cases a hospital bill isn’t actually a bill, but essentially an instrument in a complex negotiation between insurers and caregivers, with bewildered patients stuck in the middle. It’s difficult to know which charges are real and which ones aren’t, and which bills to pay and which ones to ignore. It’s one reason why medical debt is a huge factor in so many bankruptcies.

Hospital bills that amount to legal fictions certainly don’t help consumers keep themselves out of debt trouble. Todd Fassler’s bill is a perfect example — he left the hospital on July 9, 2015. His bill said his $153,000 payment was due by July 27.

 

This $153,000 rattlesnake bite is everything wrong with American health care – The Washington Post.

he assortments of bacteria that live within the intestines of isolated tribes are far more diverse than the microbes found in the guts of Americans — and scientists say such findings have implications for modern-day maladies ranging from obesity to antibiotic resistance.

The latest studies into the varying genetic signature of microbes found in the intestinal tract — also known as the microbiome — focus on Yanomami Indians in Venezuela’s Amazon region as well as on Papua New Guineans. The studies were published this week by Science Advances and Cell Reports, respectively.

In both cases, researchers found a greater variety of bacterial species than is commonly found in industrial societies.

 

“These findings suggest that lifestyle practices that reduce bacterial dispersal — specifically, sanitation and drinking water treatment — might be an important cause of microbiome alterations,” the University of Alberta’s Jens Walter, senior author of the Papua New Guinea study, said in a news release.

Sanitation practices are generally a good thing, but scientists say beneficial bacteria are lost along the way. For example, the team behind the research in Venezuela found that the Yanomami tribespeople harbored bacteria that may play a role in boosting immune response and metabolizing carbohydrates. Another example is Oxalobacter formigenes, a microbe that’s linked to a decreased risk of kidney stones.

“The challenge is to determine which are the important bacteria whose function we need to be healthy, and have a healthy, educated immune system and a healthy metabolic system,” said Maria Dominguez-Bello, a medical researcher at New York University’s Langone Medical Center who is the senior author of the study.

‘Alarming’ antibiotic resistance

Dominguez-Bello and her colleagues also found that the Yanomami tribespeople, who were “uncontacted” by Western visitors until 2009, nevertheless had gut bacteria with genes that could activate resistance to antibiotics. Some of the resistance genes could counter even the third- and fourth-generation synthetic antibiotics created to fight modern diseases.

The researchers say their findings imply that bacteria may possess an ancient but complex set of defense mechanisms that swing into action whenever they come across new threats.

Co-author Gautam Dantas, an immunologist at Washington University School of Medicine, told reporters that the finding was “alarming to us.”

“It emphasizes the need to ramp up our research for new antibiotics, because otherwise we’re going to lose this battle against infectious diseases,” Dantas told reporters.

The gut bacteria were extracted from fecal samples as well as skin swabs and mouth swabs, and then subjected to genetic analysis. The fact that the microbial communities were more diverse is in line with previous studies that have focused on Hadza hunter-gatherers in Tanzania and the Matses people of the Peruvian Amazon.

What’s good for the gut

The microbiome has become a topic of increasing interest in recent years, because scientists suspect it plays a crucial role in human health. The best-known illustration of the microbiome’s importance is the use of “fecal transplants” to cure a life-threatening intestinal infection known as C. difficile. In the future, microbiome therapy could address autism, obesity, food allergies and immune deficiencies.

Some of the bacteria identified in the guts of the Yanomami “might have therapeutic value” for such conditions, said Jose Clemente from the Icahn School of Medicine at Mount Sinai, another co-author of the study.

Dominguez-Bello emphasized that microbiome studies could help the Yanomami as well as more industrialized societies.

“It seems inevitable that the world is converging to westernized lifestyles,” she told reporters, “and so far it has been inevitable to observe how Amerindians when they integrate, or Africans when they westernize — how they quickly suffer our current diseases, obesity, diabetes. So I think that by learning what went wrong with our lifestyle … we’ll also benefit them in not suffering the same health consequences.”

In addition to Dominguez-Bello, Dantes and Clemente, the authors of the Science Advances study, “The Microbiome of Uncontacted Amerindians,” include Erica Pehrsson, Martin Blaser, Kuldip Sandhu, Zhan Gao, Bin Wang, Magda Magris, Glida Hidalgo, Monica Contreras, Óscar Noya-Alarcón, Orlana Lander, Jeremy McDonald, Mike Cox, Jens Walter, Phaik Lyn Oh, Jean Ruiz, Selena Rodriguez, Nan Shen, Se Jin Song, Jessica Metcalf and Rob Knight.

In addition to Walter, the authors of the Cell Reports study, “The Gut Microbiota of Rural Papua New Guineans: Composition, Diversity Patterns and Ecological Processes,” include Inés Martínez, James Stegen, Maria Maldonado-Gómez, A. Murat Eren, Peter Siba and Andrew R. Greenhill.

Microbiome Marvels: Tribes’ Gut Bacteria Reveal Biological Surprises – NBC News.com.

Anti Angiogenesis Foods

 

This is your last chance, after this there is no turning back.
You take the blue pill, the story ends; you wake up in your bed, and believe whatever you want. You take the red pill, you stay in wonderland and I show you how deep the rabbit hole goes.I know you’re out there, I can feel you now.
I know that you’re afraid of us…You’re afraid of change. I don’t know the future, I didn’t come here to tell you how this will all end…I came here to tell you how it’s going to begin

Clip3.jpg JPEG Image, 464 × 329 pixels.

 

At first, follicles release inflammatory proteins, or cytokines, which alert the immune system to a wound, researchers say.

The immune system responds by sending macrophages to the problem area. Macrophages are white blood cells that engulf and devour pathogens, but they also release cytokines that can trigger a variety of responses in cells, such as causing them to proliferate.

In this particular situation, macrophages will secrete signaling molecules called tumor necrosis factor alpha, which at certain levels, will prompt plucked and unplucked follicles to grow hair.

A robust regenerative response seemed to be dependent on the density of signaling behaviors, researchers said.

For example, when researchers plucked mouse hairs in a diffuse pattern, in an area with a diameter larger than 6 millimeters, no hairs regenerated.

But when a dense concentration of hairs were pulled in an area 3 to 5 millimeters across, the plucked hairs grew back and new hairs sprouted nearby.

“The quorum sensing circuit we describe here provides a way for injured hair follicles to collectively assess the magnitude and extent of injury that the skin has sustained and make an all-or-none decision whether or not to regenerate.”

Researchers discover trick to regrowing lost hair – LA Times.

Norsworthy has been in prison since 1987, serving a life sentence for second-degree murder. (S) he has twice delayed scheduled parole hearings in recent months.

 

SACRAMENTO, Calif. —A federal judge on Thursday ordered California’s corrections department to provide a transgender inmate with sex change surgery, the first time such an operation has been ordered in the state.

U.S. District Court Judge Jon Tigar in San Francisco ruled that denying sex reassignment surgery to 51-year-old Michelle-Lael Norsworthy violates her constitutional rights. Her birth name is Jeffrey Bryan Norsworthy.

The ruling marks just the second time nationwide that a judge has issued an injunction directing a state prison system to provide the surgery, said Ilona Turner, legal director at the Transgender Law Center in Oakland, which helped represent Norsworthy.

The previous order in a Massachusetts case was overturned last year and is being appealed to the U.S. Supreme Court.

In his ruling in California, Tigar said the surgery has actually been performed just once on an inmate, an apparent reference to a person who castrated himself in Texas then was given the surgery out of necessity.

Norsworthy, who was convicted of murder, has lived as a woman since the 1990s and has what Tigar termed severe gender dysphoria — a condition that occurs when people’s gender at birth is contrary to the way they identify themselves.

“The weight of the evidence demonstrates that for Norsworthy, the only adequate medical treatment for her gender dysphoria is SRS,” Tigar wrote, referring to sex reassignment surgery.

California Department of Corrections and Rehabilitation officials said they are considering whether to appeal the ruling.

“This decision confirms that it is unlawful to deny essential treatment to transgender people” in or out of prison, said Kris Hayashi, executive director of the Transgender Law Center. “The bottom line is no one should be denied the medical care they need.”

If the order stands, Norsworthy would be the first inmate to receive such surgery in California, said Joyce Hayhoe, a spokeswoman for the federal receiver who controls California prison medical care.

Hayhoe said it’s not known how much the surgery would cost, but it could run as high as $100,000, depending on the circumstances.

Corrections officials, in previous court filings, argued that Norsworthy has received proper medical and mental health care for more than 15 years and is in no immediate medical danger if the surgery is not performed.

Her care included counseling, mental health treatment and hormone therapy that the department said “has changed her physical appearance and voice to that of a woman” while helping her find her gender identity.

That care is consistent with what other judges nationwide have found to be appropriate for transgender inmates, the department said.

Norsworthy has been in prison since 1987, serving a life sentence for second-degree murder. She has twice delayed her scheduled parole hearings in recent months.

She currently is housed at Mule Creek State Prison, an all-male prison in Ione, 40 miles southeast of Sacramento.

The sex change surgery would prompt practical problems, the department said.

It said keeping Norsworhy in a men’s prison could invite violence, including possible assault and rape.

But she could also face danger at a women’s prison – or pose a threat herself – because she had a history of domestic violence before her murder conviction, the department said.

Last month, attorneys for the transgender inmate convicted of murder in Massachusetts asked the U.S. Supreme Court to overturn a ruling denying her request for sex reassignment surgery.

A federal judge in 2012 ordered the Massachusetts Department of Correction to grant the surgery to Michelle Kosilek, but the ruling was overturned in December by the 1st U.S. Circuit Court of Appeals.

As in California, the appeal in Massachusetts cited security concerns about protecting the inmate.

Courts in other states have ordered hormone treatments, psychotherapy and other treatments but not surgery.

 

via Judge orders California to pay for inmate’s sex change | News – KCRA Home.

A 1,000-year-old Anglo-Saxon salve of onion, garlic, and part of a cow’s stomach could potentially eradicate the MRSA superbug problem.

An Anglo-Saxon Expert, Christina Lee, of the University of Nottingham, spotted the eye infection remedy in a medical volume called Bald’s Leechbook that was held in the British Library in London. According to CBS News, it is one of the earliest known textbooks in medicine dating back to the 10th Century.

When they tested the mixture to the superbug Methicillin-resistant Staphylococcus aureu (MRSA) in lab-cultured Petri dishes and on the infected wounds of mice, they found that it killed 999 out of 1,000 bacterial cells.

 

“We thought that Bald’s eye salve might show a small amount of antibiotic activity, because each of the ingredients has been shown by other researchers to have some effect on bacteria in the lab,” says Freya Harrison, one of the lead Nottingham researchers, in a statement. “But we were absolutely blown away by just how effective the combination of ingredients was.”

 

Lee and her team were “astonished” as they studied the results of their experiments. She believes that modern research can learn more from past knowledge and findings that are found in old world tomes and medical books.

 

“When we built this recipe in the lab I didn’t really expect it to actually do anything,” microbiologist Steve Diggle said, noted CBS. “When we found that it could actually disrupt and kill cells in S. aureus biofilms, I was genuinely amazed. Biofilms are naturally antibiotic resistant and difficult to treat so this was a great result.”

 

The group of researchers recreated the 10th century concoction according to its original instructions. The text was translated from Old English, and in the recipe they made use of two species of Allium (garlic and onion or leek), wine, and bile from a cow’s stomach. They let the potion brew in a brass pot, then strained through a cloth, and left to sit for nine days, according to ABC News.

 

The AncientBiotics team at the University of Nottingham is currently looking for more funding to further their research. The preliminary study was presented at the conference of the Social for General Microbiology in Birmingham in England last Wednesday.

According to the U.S. CDC, the MRSA has become a major problem in the healthcare industry. It affects around 90,000 Americans yearly and has already costed billions of dollars. The MRSA kills off around 20,000 people each year.

The end of doctors:

 

Hospital admissions plummeted at Vidant Health-operated facilities after it started remotely monitoring patients suffering from heart failure, diabetes and high blood pressure. Patients at the University of Pittsburgh Medical Center send doctors pictures of their wounds from offsite in order to find out if they need antibiotics. And a patient at the hospital had his blood-pressure medication adjusted after he wirelessly sent the doctors biometrics from his home.

Such stories, which were just chronicled in The Wall Street Journal, are becoming increasingly common. Remote monitoring can empower patients and lead to more fine-tuning of care while saving hospitals money and helping hospitals meet their Affordable Care Act (ACA) goals, like reducing hospitals readmissions (or else facing a reduction in Medicare reimbursement).

 

Sales of remote-monitoring devices are expected to top $32 billion this year, according to Market Research Group, and will grow at a compounded annual growth rate of 9.2% between 2014 and 2019.

 

 

MRSA Superbug Killed by 1000-Year-Old Medieval Eye Infection Treatment : Latinos Health News : Latinos Health.

TaurineTaurine plays a major role in good liver function via detoxification and the formation of bile. Inadequate levels of taurine are common in many patients with chemical sensitivities and allergies. Taurine is the major amino acid required by the liver for the removal of toxic chemicals and metabolites from the body. Impaired body synthesis of taurine will reduce the ability of the liver to detoxify environmental chemicals such as chlorine, chlorite bleach, aldehydes produced from alcohol excess, alcohols, petroleum-based solvents and ammonia. Recent findings are demonstrating, that taurine is one of the major nutrients involved in the body’s detoxification of harmful substances and drugs, and should be considered in the treatment of all chemically sensitive patients. Taurine is helpful for high blood cholesterol and gall bladder problems, alcohol withdrawal, hepatitis and jaundice.

via Positive Health Online | Article – A Healthy Liver and Weight Loss.

“IT’S like coffee times ten,” raves one enthusiast. “I use it a couple of times a week and problems solve themselves. At the end of the day, I haven’t wasted hours on frivolous websites. At the end of the week, my apartment is clean.” This marvel of productivity is not a new energy drink or an experimental wonder drug but a simple electrical device that he built at home for less than $10. Whenever this physicist feels like an extra burst of motivation, he places electrodes on his skull and sends a jolt of electricity into his brain.

The currents, which are typically applied for ten to 20 minutes, are hundreds of times smaller than the seizure-inducing shocks used in electroconvulsive therapy. Plans to make such transcranial direction current stimulation (tDCS) machines are freely available online and their components can be bought at hobbyist stores. Kits cater to those lacking soldering skills, and now companies are emerging offering nicely designed and packaged brain zappers for mainstream consumers.

Not everyone using tDCS is seeking to become more efficient in their daily life. Some hope to enhance their concentration for study or video gaming; others want to boost their memory, speed up learning or induce meditative calm. Yet more are trying to self-medicate for conditions such as depression, chronic pain and motor, sensory or neurological disorders. The benefits might sound implausible, but there is some science to support them. The idea goes back a long way. Scribonius Largus, a first-century Roman physician, prescribed the shock of an electric ray for headaches, and in the 19th century electrical pioneers such as Luigi Galvani and Alessandro Volta toyed with crude bioelectric experiments. It was not until the 1960s, however, that the first rigorous studies of electrical brain stimulation took place.

Directing the flow

The theory behind tDCS is that a weak direct current alters the electric potential of nerve membranes within the brain. Depending on the direction of the current, it is said to make it easier or more difficult for neurons in a brain circuit to fire. Position the electrodes correctly and choose the right current, so the idea goes, and you can boost or suppress all kinds of things. Some researchers have reported that tDCS can reduce pain, ease depression, treat autism and Parkinson’s disease, control cravings for alcohol and drugs, repair stroke damage, and accelerate recovery from brain injuries, to say nothing of improving memory, reasoning and fluency. Remarkably, some effects seem to persist for days or even months. And the closer that scientists look at tDCS, the more they seem to find. Scientific papers about the technology appear at an ever-faster rate.

Hardly surprising, then, that DIY brain hackers want in on the action. Christopher Zobrist, a 36-year-old entrepreneur based in Vietnam, is one of them. With little vision he has been registered as blind since birth due to an hereditary condition of his optic nerve that has no established medical treatment. Mr Zobrist read a study of a different kind of transcranial stimulation (using alternating current) that had helped some glaucoma patients in Germany recover part of their vision. Despite neither the condition nor the treatment matching his own situation, Mr Zobrist decided to try tDCS in combination with a visual training app on his tablet computer. He quickly noticed improvements in his distance vision and perception of contrast. “After six months, I can see oncoming traffic two to three times farther away than before, which is very helpful when crossing busy streets,” he says.

Online communities dedicated to tDCS are full of similar stories. More still claim to have gained cognitive enhancements that give them an edge at work or play. Users follow the latest scientific papers avidly and attempt to replicate the results at home, discussing the merits of different currents, waveforms and “montages” (arrangements of the electrodes on the skull).

Happiness and health may always be more than just a 9-volt battery away

Dissenting voices are rare. Here and there are tales of people who experienced headaches, nausea, confusion or sleeplessness after tDCS, while temporary visual effects and mild skin burns are fairly common. There have been no reports of seizures, serious injuries or deaths. But that does not mean it is without risk, says Peter Reiner, co-founder of the National Core for Neuroethics at the University of British Columbia. He says DIY users may place electrodes incorrectly, thus stimulating the wrong part of their brain, or reverse the polarity of current, potentially impairing the very things they are trying to improve. No one really knows how tDCS interacts with chemical stimulants or recreational drugs like marijuana, or with pre-existing conditions like epilepsy. Even something as fundamental as being left-handed can alter the functional organisation of the brain. And if the benefits of tDCS can persist for weeks, perhaps its side-effects can linger, too. Many neuroscientists are particularly worried that the use of tDCS by children and young adults could affect their long-term neural development.

Some of these concerns can be addressed by manufacturing tDCS devices to make it difficult, or impossible, to exceed recommended currents or to apply the electrodes incorrectly. One such product already exists. The Foc.us V2, made by Transcranial, a London company, is advertised as a $199 pocket-sized controller that pairs with a $99 headset intended to help with concentration and reaction speed while videogaming. Donning the headset automatically positions the electrodes on the left and right temples, and both the duration and maximum current are capped. A second headset provides a different montage aimed at improving performance and motivation while exercising.

In reality, however, there is no guarantee that even slick products are any safer than a pocket-money brain stimulator assembled at home from a 9-volt battery, electrodes, a few wires and other components. Unlike the tDCS machines used for medical trials and clinical research, consumer versions may not have been assessed by any official body for safety or effectiveness. If the maker insists they are for use only by healthy adults to enhance cognition or leisure activities and make no diagnostic or therapeutic claims, such “wellness” devices have slipped under the regulatory radar of both the Medical Devices Directive in Europe and the Food and Drug Administration (FDA) in America.

That worries some experts. A recent paper from the Institute for Science and Ethics at the University of Oxford points out that consumer tDCS products are mechanically and functionally equivalent to medical neurostimulation devices that require licensing. Why regulate the version that is likely to be operated responsibly by health professionals, and not the one freely available to unskilled and inexperienced users? The Nuffield Council on Bioethics agrees, recommending in 2013 that the European Commission should consider regulating all such gadgets under its medical devices regime, regardless of the purposes for which they are marketed.

The Institute for Science and Ethics proposes a graded regulation system that errs on the side of consumer choice for tDCS devices, requiring comprehensive, objective information about risks and benefits to allow users to make informed decisions. But it wants supplying brain zappers to children to be made illegal. Last year the FDA allowed transcutaneous electrical nerve stimulator (TENS) machines for headache relief as it rated them as low-to moderate-risk devices. TENS devices use a different waveform to tDCS and target cranial nerves rather than the brain itself, but they rely on a similar controller and head-mounted electrodes. Before allowing new TENS products to be sold, the FDA now wants to see evidence that the components are not likely to cause injury, that the controller can reliably provide the correct output, that there are no thermal or mechanical hazards, and that clinical data demonstrate the device is safe and effective as a headache treatment. Recent draft FDA guidelines for wellness devices suggest tDCS machines may eventually be regulated in a similar way.

Going underground

The University of British Columbia’s Dr Reiner doubts that any manufacturer today can provide such information for tDCS. Even if they could, the cost of gathering it would make consumer devices more expensive. “When you can make a tDCS device yourself for less than $20, we would advise strongly against heavy regulation because it will only drive the technology underground,” he says.

Proving the effectiveness of brain stimulation will be difficult. Although it may well do something, exactly what is open to question. As the hype around tDCS grows, some neuroscientists are starting to question whether the technology really is the panacea it appears to be.

In 2013 Teresa Iuculano and Roi Cohen Kadosh of the Department of Experimental Psychology at the University of Oxford split volunteers up into three groups and asked them to learn a made-up mathematical notation system. The first two groups received tDCS to different parts of the brain previously associated with numerical understanding and learning, while a non-functional “sham” device was used on the third group as a control. After a week, all three groups were tested on how well they had learned the new notation system, and whether they could use it in practice. The first group showed an improvement in learning compared with the control group, but a decrease in their ability to apply their knowledge, while the second group experienced the opposite result. This suggests that the brain is actually rather well balanced: boost performance in one cognitive realm through stimulation, and aptitude in another will naturally diminish.

There is also the possibility that a variation in individual responses to tDCS will overshadow any general effects. In a study published last year, Dr Cohen Kadosh set up two groups: one of people who were anxious when presented with mathematical problems, and another who had confidence in their ability to breeze through numerical quizzes. When treated with tDCS to their prefrontal cortices, the nervous individuals improved their reaction time on simple arithmetical problems and showed reduced levels of stress. Given the same treatment, the confident group had longer reaction times and no less stress. “If you can get exactly the opposite results with a different population, that shows DIY brain hackers and companies marketing stimulation to improve gaming or other abilities are not on the right track,” says Dr Cohen Kadosh. “We need to understand how the brain works in different people.”

Felipe Fregni, director of the Laboratory of Neuromodulation at Harvard Medical School, says tDCS has been shown to accelerate the learning of new skills. But he agrees that individual variation is important, noting that younger people sometimes do not improve as much as older subjects, and that people at later stages of learning may even experience detrimental effects. “The more science you know, the more confused you can become of what really is the effect of tDCS,” says Dr Fregni.

One advantage of the deluge of scientific papers is that they can be subjected to meta-analysis, whereby studies can be statistically combined to tease out new discoveries. Last year, Jared Horvath, a neuroscientist at the University of Melbourne in Australia, published a meta-analysis of 30 measurements taken during tDCS studies, including neural responses, oxygen levels and electrical activity in the brain. Surprisingly, he found that tDCS had a reliable effect on only one: the electrical response of muscles to stimulus, and even that has steadily declined in studies over the last 14 years. Mr Horvath believes this indicates that the response has historically been measured poorly and that it too will eventually disappear as techniques mature.

Equally troublesome is a meta-analysis of the cognitive and behavioural effects on healthy adults that Mr Horvath subsequently carried out. As before, he included only the most reliable studies: those with a sham control group and replicated by other researchers. It left 200 studies claiming to have discovered beneficial effects on over 100 activities such as problem solving, learning, mental arithmetic, working memory and motor tasks. After his meta-analysis, however, tDCS was found to have had no significant effect on any of them.

If tDCS alters neither the physiology of the brain nor how it performs, thinks Mr Horvath, then evidence suggests it is not doing anything at all. Marom Bikson, a professor of biomedical engineering at City University of New York, disagrees. “I can literally make you fall on your butt using the ‘wrong’ type of tDCS,” he says. Dr Bikson thinks the biggest challenge for tDCS is optimising techniques, such as the dose.

Mr Horvath notes that many papers measure 20 or more outcomes, with brain stimulation showing a weak effect on one or two. “But in the title and abstract, that’s all they talk about,” he says. “No one mentions the tons of effects that tDCS didn’t have an impact on but that technically it should have if it is doing what the researcher thinks it is.”

Another problem might be the small sample size, sometimes as few as ten or 15 people. Mr Horvath says future studies should use at least 150 subjects. There is, of course, the possibility that Mr Horvath’s analyses are flawed. His paper included only one-off sessions, while many scientists believe the effects of tDCS accumulate with repetition. However, too few multiple-session studies exist for a valid meta-analysis. Dr Cohen Kadosh points out that individual variations could make the technology look as though it is doing nothing when in fact it has real but opposing effects in different people. Mr Horvath insists that his analysis allows for this possibility.

Critics might also wonder why Mr Horvath omitted tests where tDCS seems to have been most effective, in alleviating, for instance, clinical conditions such as depression. He admits that would be useful but says, “If something doesn’t demonstrate any type of effect in healthy people, it becomes incredibly difficult, if not impossible, to argue why it would work within a clinical population.”

Not all neuroscientists are defending the status quo. “I’m not surprised that he found no effect from conventionally applied tDCS,” says Jamie Tyler, a professor at Arizona State University and one of the founders of Thync, a Silicon Valley startup that recently unveiled a smartphone-controlled tDCS device. Thync tried to replicate some basic tDCS findings on cognition but could not do so. Dr Tyler now believes that tDCS may not directly stimulate the brain at all but instead modulates cranial nerves in the skull, like the headache-busting TENS technology. He designed the Thync device, a pocket-sized unit with disposable pre-shaped electrodes, to target these nerves with the aim of generating either relaxed or energetic mental states.

A shot of caffeine

Dr Tyler recently published a study of 82 people with a control. Its results suggest that Thync’s device can reduce psychophysiological stress by altering skin conductivity (a measure used in pseudoscientific lie detectors), stress enzymes and heart rate variability. He likens Thync’s “modified tDCS” programs to ingesting either a third of a cup of coffee or a glass of wine, and says no effect has been found on cognitive processes like working memory. While Thync’s stimulator is not yet available to the public, the firm was willing to give your correspondent a pre-launch trial.

The Thync device attaches with one sticky electrode on the right temple and one behind the right ear. The unit is controlled via a smartphone app, with the user able to adjust the intensity but not the duration of the session. At first, the unit generated a barely perceptible crawling feeling on the skin near the electrodes, building gradually to a pronounced tingling sensation. Over the 20-minute session, the strength of the signal varied up and down according to a preset routine. It felt itchy at times and, at its most powerful, caused muscles in the forehead to spasm alarmingly. Although the experience was not altogether unpleasant, any extra energy or focus proved, alas, elusive. Dr Tyler acknowledged that perhaps one in four people do not perceive any immediate benefit from the device.

Even for those who find themselves susceptible to its charms, the challenges for a product like Thync are formidable. The cognitive enhancements of a strong cup of tea or a glass of vintage Burgundy are well established. And partaking of them can be socially acceptable, deliciously enjoyable and rapidly achieved. None of these can be said of a disconcerting gizmo that needs half an hour to work and causes eyebrows to raise, both literally and socially.

Regardless of their questionable utility and effectiveness, tDCS gadgets are too novel, cheap and alluring to simply dismiss. Consumer-wellness devices like Thync may appeal to those who cannot use caffeine or alcohol for medical or religious reasons, and there will always be healthy overachievers seeking to supercharge their cognition for study or work. More importantly, tDCS presents the tantalising promise of relief from some medical conditions for which traditional therapies are either ineffective or unaffordable. As the University of Melbourne’s Mr Horvath says, “If there are ten percent of people who are feeling a huge effect, even if that’s placebo, who are we to say no to them?”

If people want to experiment with tDCS, there seems to be no reason to prevent them, provided it is done in the safest way possible. Devices could be regulated lightly with a view to safety rather than effectiveness, and neuroscientists encouraged to design future studies with more rigour. Happiness and health may always be more than just a 9-volt battery away, but brain hacking looks like it is here to stay.

Neurostimulation: Hacking your brain | The Economist.

If it was some africian girl getting part of her sex organs cut off there would be killings in the street. But cut off some little white boys dick and all is well!

Proof there is something in the water in flordia that makes all their brains just fucking turn upside down.

DELRAY BEACH — In what is believed to be a first in the nation, a Palm Beach County judge on Tuesday ordered the arrest of a Boynton Beach woman because she refuses to have her son circumcised.

Circuit Judge Jeffrey Gillen signed the arrest warrant for 30-year-old Heather Hironimus after telling her attorney Thomas Hunker that her concerns about her 4-year-old son’s welfare were irrelevant.

“Is she in the courtroom or not?” the judge asked.
hand holding scalpel photo

Explaining that Hironimus and her son had sought refuge at an unspecified domestic violence shelter to block the boy’s father from carrying out plans for the circumcision, Hunker admitted she had decided not to attend the hearing.

Her absence defied an order Gillen signed Feb. 25, ordering Hironimus to bring the boy to court on Friday to be turned over to his father, Dennis Nebus. When she didn’t appear, Gillen gave her until 2 p.m. Monday to appear in court with her son.

Exactly how the dispute will unfold is unclear. Once she is arrested, she will be brought before Gillen, Hunker said. It is likely the child will be turned over to Nebus. Gillen has given him authority to have the circumcision done without Hironimus’ approval.

Nebus’ attorneys declined comment. But, Hunker said, Hironimus is scared both for herself and her son.

“She doesn’t believe she should be incarcerated for protecting her child,” he said.

The strange and convoluted case has spawned international headlines and the attention of self-described “intactivists,” who are against the forced circumcision of children. Carrying signs declaring, ““His Body, His Right,” and “Whose Penis? Whose Body? Whose Rights?” about 15 activists gathered outside the South County Courthouse to support Hironimus, who has used social media to publicize her son’s plight.

The now controversial case began as a simple paternity lawsuit. Three months after his son was born in October 2010, Nebus went to court to claim his parental rights. Nebus, 47, of Boca Raton, and Hironimus never married.

In 2012, the two signed a parenting agreement. Hironimus agreed her son could be circumcised as long as Nebus paid for it.

Soon after, she had second thoughts about allowing her son to undergo the procedure. After various court hearings, Gillen in May ruled that the agreement trumped her reservations. The 4th District Court of Appeal upheld his decision without comment.

The agreement, Gillen wrote, is clear and unambiguous. “Mother agrees to and shall timely execute any and all documents reasonably necessary to effectuate the circumcision of the minor child,” he wrote, quoting the agreement.

Before reaching his conclusion, he heard from pediatric urologist Charles Flack. The physician testified that while the procedure wasn’t medically necessary it’s advisable. Penile cancer only afflicts uncircumcised men. Further, uncircumcised men are at a greater risk of contracting HIV and other sexually transmitted diseases. The procedure, Flack testified, lasts only 17 minutes and causes little discomfort.

Men who gathered outside the courthouse disagreed. Carl Silverman, who drove from his home in Royal Palm Beach to participate in the protest, said he still remembers the pain of the circumcision he had done as an infant. “From my earliest days, I remember a searing pain down there,” he said.

Mothers who gathered with their children for the protest said the operation would be particularly traumatic for Hironimus’ son, who is not being named by court order. “(He’s) 4 1/2 years old and I know he does not want this,” said Kristen Shockley, of Boynton Beach, who said she is friends with Hironimus. “He’ll remember it. I know I remember things that happened to me when I was four.”

Attorney Andrew DeLaney, who wrote a paper on female genital mutilation and male circumcision while in law school at St. John’s University and has been watching the case unfold from his home in Texas, said Gillen’s decision seems draconian given the passage of time and the mother’s current wishes.

“A reasonable judge, in my opinion, would void the agreement due to the change of circumstances and recognize that it is not in the best interests of this boy to force the mother into consenting to the procedure at this point,” he said.

But, others said, a court order is a court order. The agreement, approved by Gillen, is just that. It can’t be flaunted by those who signed off on it and now have second thoughts.

Further, the medical community appears to be on Gillen’s side. While stopping short of mandating circumcision, the federal Centers for Disease Control and Prevention in November issued guidelines saying the benefits of circumcision far outweigh the risks. In the guidelines, it urges doctors to tell parents of males the benefits of the procedure.

Editor’s Note: An earlier version of this story incorrectly said protestors opposed the forced sterilization of children. They oppose the forced circumcision of children.

Warrant issued for Florida woman who won’t allow son’s… | www.ajc.com.

The female body: Shape-shifting

Brian Buntz

Popularized by the Russian economist Nikolai Kondratiev, long wave theory holds that decades of economic progress follow from technological breakthroughs such as was the case with the development of the steam engine, the railway, electrical and chemical engineering, automobiles, and computing technology.

In the most recent period, the microprocessor is the single most important technology, making possible everything from personal computers and smartphones, to smart bionic limbs and wireless-enabled medical devices. Indeed, much of our very culture now seems to revolve around the microprocessor.

Perhaps another technology will emerge as a key driver of medical technology in years to come. And medicine could be one of the principal industries to benefit from the next decades-long technological period, which we could be on the cusp of entering now. The Slovak theorist Daniel Smihula refers to the next decades-long phase as the post-informational technological revolution, and expects it to begin between 2015 and 2020.

A 2010 Allianz report also forecasted a wave of medical technology innovation playing a central role in the next long-term technological phase, arguing that such periods typically emerge after major financial crashes or periods of economic stagnation, and that the Great Recession may be one such example of that. Kondratiev himself believed in a long-term boom–bust cycle, asserting that the Great Depression would not spell the end of capitalism but give rise to a new period of economic success in the West. Stalin apparently disagreed and had the theorist shot by a firing squad.

Whether long-wave theorists are right about the early 21stcentury giving rise to another technological megacycle, there is a definite need for a new wave of innovation in healthcare—in part because the world’s graying population. By 2050, the population percentage in the United States that is over 65 stands to roughly double—and nearly triple in Asia and Latin America. Add to that growing pressures to contain healthcare costs and an uptick in chronic diseases, and we’ve got a big problem on your hands.

If Kondratiev’s grand vision is true, there is a good chance that much of the prognosticating about the future of medical technology will seem myopic by comparison. For one thing, a lot of projections about healthcare’s future are based on applications of electronics. And while electronics will undoubtedly play an integral in an ever-widening number of medical technologies, long-wave theory holds that one technology revolution lays the groundwork for the next. So it is possible that the innovation made possible by electronics could give rise to other technological fields that would characterize the next era. Contenders could include fields like nanotechnology, genomics, biotechnology, or 3-D printing, any of which may ultimately catalyze a wave of long-term medical innovation.

Such a shift may be already underway. The Economist just penned an article stating that the U.S. healthcare system is a “wasteful and inefficient industry, is in the throes of great disruption.” Similar upheaval can be seen elsewhere.

Perhaps revolution is a good word to describe the next period of technological evolution in medicine. While there is clearly a need for novel devices that make healthcare more precise and efficient, any new technology that threatens entrenched medical business models must battle against those who would preserve the status quo.

Economist article below:

THE best-known objective of America’s Affordable Care Act of 2010—commonly known as Obamacare—was