There is a nanomotor in your mitochondria thats spins 9k rpms, thats faster than a corvet

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ATM I'm reading this book "Dirt To Soil". It's mostly about soil improvement and it also stresses the importance of a variety of flowers as part of your cover crops to attract pollinaters and also predator insects to keep pests in check. That might be an option to reduce the fly problems naturally.

It's a very interesting read anyway, in case you're interested.

yuge gabe brown fan! guy is an innovator and great inspiration to farmers everywhere. his story is a great.
I'll add the book to my list thanks!
 

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That's all find and dandy, but now tell me how midichlorians work. When people are suffering and dying, is that god's work also or are you going to blame that on a scapegoat? You wouldn't have to cure cancer if an all perfect god created life. All this shows is that evolution is beyond your ability to reason and that you evolved from a plant.
I'll get back to you on midichlorians later, its not in my field of view at the mo.

weather you think we evolved from a plants or not wasn't in my field of view either, the idea that at the cellular levels we are a few ions separated is mind boggling to me, a simple farmer. plants create oxygen as they grow, we burn oxygen as we grow, we are in need of each other for healthy life, that's what interests me.
the light in each is the spark of God.

1 In the beginning God created the heaven and the earth.


2 And the earth was without form, and void; and darkness was upon the face of the deep. And the Spirit of God moved upon the face of the waters.


3 And God said, Let there be light: and there was light.


4 And God saw the light, that it was good: and God divided the light from the darkness.


5 And God called the light Day, and the darkness he called Night. And the evening and the morning were the first day.


6 And God said, Let there be a firmament in the midst of the waters, and let it divide the waters from the waters.


7 And God made the firmament, and divided the waters which were under the firmament from the waters which were above the firmament: and it was so.


8 And God called the firmament Heaven. And the evening and the morning were the second day.


9 And God said, Let the waters under the heaven be gathered together unto one place, and let the dry land appear: and it was so.


10 And God called the dry land Earth; and the gathering together of the waters called he Seas: and God saw that it was good.


11 And God said, Let the earth bring forth grass, the herb yielding seed, and the fruit tree yielding fruit after his kind, whose seed is in itself, upon the earth: and it was so.


12 And the earth brought forth grass, and herb yielding seed after his kind, and the tree yielding fruit, whose seed was in itself, after his kind: and God saw that it was good.


13 And the evening and the morning were the third day.


14 And God said, Let there be lights in the firmament of the heaven to divide the day from the night; and let them be for signs, and for seasons, and for days, and years:


15 And let them be for lights in the firmament of the heaven to give light upon the earth: and it was so.


16 And God made two great lights; the greater light to rule the day, and the lesser light to rule the night: he made the stars also.


17 And God set them in the firmament of the heaven to give light upon the earth,


18 And to rule over the day and over the night, and to divide the light from the darkness: and God saw that it was good.


19 And the evening and the morning were the fourth day.


20 And God said, Let the waters bring forth abundantly the moving creature that hath life, and fowl that may fly above the earth in the open firmament of heaven.


21 And God created great whales, and every living creature that moveth, which the waters brought forth abundantly, after their kind, and every winged fowl after his kind: and God saw that it was good.


22 And God blessed them, saying, Be fruitful, and multiply, and fill the waters in the seas, and let fowl multiply in the earth.


23 And the evening and the morning were the fifth day.


24 And God said, Let the earth bring forth the living creature after his kind, cattle, and creeping thing, and beast of the earth after his kind: and it was so.


25 And God made the beast of the earth after his kind, and cattle after their kind, and every thing that creepeth upon the earth after his kind: and God saw that it was good.

more at https://www.biblegateway.com/passage/?search=Genesis+1&version=KJV

god made water....light... soil...grass...rain...cattle all in the beginning, all are essential to a full great life.

I'm here reveling in it and bringing the good news to doubters and some of the lost that have ears to hear.
 

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Abstract
Mitochondria play crucial roles in regulating metabolism and longevity. A body of recent evidences reveals that the gut microbiome can also exert significant effects on these activities in the host. Here, by summarizing the currently known mechanisms underlying these regulations, and by comparing mitochondrial fission–fusion dynamics with bacterial interactions such as quorum sensing, we hypothesize that the microbiome impacts the host by communicating with their intracellular relatives, mitochondria. We highlight recent discoveries supporting this model, and these new findings reveal that metabolite molecules derived from bacteria can fine‐tune mitochondrial dynamics in intestinal cells and hence influence host metabolic fitness and longevity. This perspective mode of chemical communication between bacteria and mitochondria may help us understand complex and dynamic environment–microbiome–host interactions regarding their vital impacts on health and diseases.

the human genome(20,000 and 25,000 genes. ) is rather small compared to the genome of the gut bacteria, virome, fungi, parasites.(3.3 million nonredundant microbial genes )It has been estimated that the microbes in our bodies collectively make up to 100 trillion cells, ten-fold the number of human cells, and suggested that they encode 100-fold more unique genes than our own genome

if I have Laszlo Boras right, our mitochondria spin one way producing metabolic water for our cells and ATP to run our systems, some? of the gut biomes mitochondria run the opposite direction doing something with the protons that our human mitochondria does the other way.
why this might matter is how many of each are doing the spinning could determine if you are going to be sick or not and why you might be sick.
what is your yuge gut biome made of compared to the relatively small genome of your "human" mitochondria.
Boras will tell you that the human mitochondria enzymatic reactions create depleted deuterium cellular water, and some of the gut bacterias will actually add to the deuterium load? https://cdn.onb.it/2020/02/Boros.pdf

when the physics and microbiology gets drilled down to the quantum levels...muh head spins trying to grasp the details God worked out...I mean random selections? junkyard tornado? hell nah!

I'm going out to look at sunrise feed my systems some blue green red and purple light!
 

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I did a search for jack.kruse last week and didnt find anything.

His talks blow me away!

Structured water is from light sunlight.

Blocking people from sunlight makes people stupid and obedient!

Robert o becker.
Many books you should get and read.
Gilbert ling.
4th phase of water.
The body electric
Many more.

 

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A separate study by Japan Crush found that 30% of unmarried men had never dated a woman at all.


Blue light destroys leptin, wrecks melatonin production, wrecks sex hormones production...blue light is out of balance.



The energy we get and use from the sun has changed us dramatically.

Artificial lights EMF and our use of electron transport chain has been altered...

Biggest users of mitochondria in our bodies , heart and brain...biggest disease vector, heart...brain is catching up in terms of diseases....
Fuck up your mitochondria and you are fooked...
 

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Should a section of the genome be naturally died off to strengthen the earth's populations?

Should stupid and sick be propagated?

Should a vax be made for every malady?

Why when you can be an abortion proponent, practitioner, why is every weak sick stupid person on earth worth extraordinary measures? that saving them with a vax or antibiotic in the long term wrecks our ability to walk outside without a mask.
'Ape and Essence' by Aldous Huxley deals with all of this in a very readable novel.
 

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Dr boras explains iron in hemoglobin, how it reacts to electromagnetism 17:30




References sited in description.
 

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Gerald Pollack's talks about water and deuterium and photosynthesis, mitochondrial health.

We are light beings.

Genesis 1:3
And God said, β€œLet there be light,” and therewas light.
 

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Selective loss of mitochondrial genome can be caused by certain unsaturated fatty acids

Abstract
Various unsaturated fatty acids had different effectiveness for maintaining the continued replication of functional mitochondria in an unsaturated fatty acid auxotroph of Saccharomyces cerevisiae (KD115). Certain isomers of octadecenoic acid (i.e., cis-9) and eicosatrienoic acid (i.e.,cis-8,11,14) permitted continued replication of mitochondria and provided cultures that contained only 4 to 5% cells that formed petite colonies. On the other hand, cultures grown with cis-12- or cis-13-octadecenoic acid or cis-11,14,17-eicosatrienoic acid, produced a 12- to 16-fold greater frequency of petite mutants (50–60%) after 8 to 10 generations of growth. The production of the petite mutants occurred despite adequate incorporation of these unsaturated fatty acids into cellular phospholipids and an apparently normal ability to undergo the initial steps in the induction of cellular respiration. The evidence suggests that some cellular processes necessary for continued mitochondrial replication depend on the structural features of the fatty acyl chains as well as the overall content of unsaturated fatty acids in membrane phospholipids. Impairment of that process by certain inadequate fatty acids or by an inadequate supply of a suitable fatty acid leads to a permanent loss of the mitochondrial genome from the cells of subsequent generations.



According to "Researchers" PUFA or linoleic acids will wreck your mitochondria. Via oxidative stress reactions.

150 years ago we ate very little seeds oils in a year, today we are soaked in it and as a consequence we are fat , stupid and sicker than ever in history.

Check out premier researcher Bill Landis.

MONDAY, DECEMBER 14, 2020
Bill Lands (aka William E.M. Lands, PhD) Resources on n-6 and n-3 PUFAs

Bill Lands is one of the titans in the field of human PUFA metabolism. As Wikipedia puts it: "...[he] is an American nutritional biochemist who is among the world's foremost authorities on essential fatty acids."
I don't call him "Bill" to be familiar, after he retired he switched from publishing as William E.M. to Bill.
Here's a select bibliography of interesting things he's written and done.
He's pretty fiery, so be sure to watch those video of his speech in (I think) 1999 at the NIH workshop, whence this quote:
Lands: (paraphrasing a soldier) "The limited n-3 and the excessive n-6 has 'screwed' the military."
β€” Tucker Goodrich (@TuckerGoodrich) August 5, 2017
It's in four parts, but it's well worth it.
Or this:
"Fifty years later, I still cannot cite a definite mechanism or mediator by which saturated fat is shown to kill people." (Lands 2008)
Graff, G., Sacks, R. W., & Lands, W. E. M. (1983). Selective loss of mitochondrial genome can be caused by certain unsaturated fatty acids. Archives of Biochemistry and Biophysics, 224(1), 342–350. https://doi.org/10.1016/0003-9861(83)90218-7
Lands, B. (2008). A critique of paradoxes in current advice on dietary lipids. Progress in Lipid Research, 47(2), 77–106. https://doi.org/10.1016/j.plipres.2007.12.001
Lands, B. (2014). Historical perspectives on the impact of n-3 and n-6 nutrients on health. Progress in Lipid Research, 55, 17–29. https://doi.org/10.1016/j.plipres.2014.04.002
Lands, B. (2020, February 1). Essential Fatty Acids Home Page. EFA Education. https://efaeducation.org/
NIH. (1999a, April 7). 1 of 4β€”Dr William Lands on Cardiovascular Disease Omega-6.
NIH. (1999b, April 7). 2 of 4β€”Dr William Lands on Cardiovascular Disease Omega-6.
NIH. (1999c, April 7). 3 of 4β€”Dr William Lands on Cardiovascular Disease Omega-6.
NIH. (1999d, April 7). 4 of 4β€”Dr William Lands on Cardiovascular Disease Omega-6.
NIH. (1999e, April 7). Workshop on the Essentiality of Omega-6 and Omega-3 Fatty Acids. https://ods.od.nih.gov/pubs/conferences/w6w3_abstracts.html
Strandjord, S. E., Lands, B., & Hibbeln, J. R. (2018). Validation of an equation predicting highly unsaturated fatty acid (HUFA) compositions of human blood fractions from dietary intakes of both HUFAs and their precursors. Prostaglandins, Leukotrienes, and Essential Fatty Acids, 136, 171–176. https://doi.org/10.1016/j.plefa.2017.03.005
Wikipedia. (2020). William E.M. Lands. In Wikipedia. https://en.wikipedia.org/w/index.php?title=William_E.M._Lands&oldid=994164694


List courtesy of Tucker Goodrich blog.
 

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DNA's sugar, deoxyribose, has five carbon atoms, which are connected to each other to form what looks like a ring. Four carbons plus an oxygen are part of the five-sided ring. The fifth carbon branches off the ring. ... The sugar in DNA is called a deoxyribose because it doesn't have a hydroxyl group at the 2' position.

 

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"...the soviets proved that the activity of the electron transport chain -the mitochondrial enzymes that extract energy from our food - is diminished not only in animals that are exposed to radio waves, but in animals exposed to magnetic fields from ordinary electric power lines."

patients with neurocirculatory asthenia cant get enough oxygen from same amount of air as normal people,- they suffer from a defect of aerobic metabolism- the radio waves and added electricity around them interferes with their mitochondrial use of oxygen...its part of the covid "flu" puzzle!...

...research included from Russian radio wave studies on humans and animals...remember how embassies are always allegedly being bombarded with microwaves to get into the CIA operatives systems?! they know how bad and dangerous electromagnetic smog is to human cellular structures...but go ahead and give your goy baby a cell phone to call you from the crib!

https://dm5migu4zj3pb.cloudfront.net/manuscripts/101000/101777/JCI46101777.pdf...paper on neurocirculatory asthenia
 
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According to publish literature all these diseases Neurocirculatory asthenia, radio waves sickness, anxiety disorder, chronic fatigue syndrome and myalgic encphalmyelitis-predispose to elevated levels of blood cholesterol and all carry an increased risk of death from heart disease. so do porphyria and oxygen deprivation. the fundamental defects in this disease of many names is that although enough oxygen and nutrients reached the cells the mitochondria the power also the cells can not efficiently use that oxygen and nutrients, and not enough energy is produced to satisfy the requirements of heart, brain, muscle and organs. this effectively starves the entire body including the heart of oxygen and can eventually damage the heart. in addition neither sugars nor fats are efficiently utilized by the cells, causing unutilized sugar to build-up in the blood leading to diabetes as well as unutilised fats to be deposited in arteries.
 

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Jack Kruse is onto something!
Light
Water
Magnetism
 

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Every animal has allotted to it a constant number of heart beats per lifetime. if it lives fast and furiously like a shrew or a mouse, it will use up its quota of heartbeats in a much shorter time than if its metabolic personality is a more temperate one
Donald r. Griffin
listening in the dark
i.gr-assets.com/images/S/compressed.photo.goodr...

calorie restriction the only method of prolong life in all animals-warm blooded and cold blooded, hibernators and non hibernators food restricted animals use less oxygen.

humans internal organs comprise less than 10% of our weight use 70% of our resting energy it is our internal organs that decide how long we live.

the engines of our lives the electron transport chain in the mitochondria... the food and oxygen combine and decide how fast these spin that speed determines our rate of living and lifespan. that speed is determined by body temp and the amount of food we eat.

another way to slow our rate of living is by poisoning the electron transport chain . using electromagnetic fields you can slow the ETC.

you can slow metabolism via electronic smog, you may live longer but ultimately way less healthy with far more metabolic syndrome and diseases...


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Outlined in his book, The Rate of Living published in 1928, Pearl conducted a series of experiments in drosophila and cantaloupe seeds that corroborated Rubner's initial observation that a slowing of metabolism increased lifespan.
 

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Here is an interesting video with a computer model showing an accurate representation of the actual DNA replication machine that's occurring right now inside your body



We have no ways to directly observe molecules and what they do -- but Drew Berry wants to change that. He demos his scientifically accurate (and entertaining!) animations that help researchers see unseeable processes within our own cells.
 

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Blue light, B12, scott kelly astronaut hyper methylation
 
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D0q5BMJV4AANOeO.jpeg

Am sunlight sets your brain up for best sleep
 

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SIVAM electronic surveillance in the amazon, they can hear a twig snap anywhere in forest? they blame "climate change" for the devastation in rain forests and Colorado's aspen forests, and the 3k year old cedars of the world...but really its pulsed RF and electromagnetic smog!

its why you have covid911



A sophisticated network of sensory equipment--including satellites, surveillance aircraft and dozens of radar systems spread across the jungle--will allow air-traffic controllers to follow clandestine flights, help isolated towns to quickly alert officials of epidemics and track environmental data. The first of those radars is scheduled to go up in Jacareacanga this month.

Colorado has a yuge network of radio communication beacons...all killing the ecosystem...its climate change alright, but its not from fossil fuel burning its from ionosphere wrecking!
 

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Doug wallace lecture. Mitochondrial etiology of common complex disease.

The boundries of reality interfere with explaining and helping to understand disease modalities.

Skip to about 17 minute mark lots of introductions you miss.
 

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keep your leg muscles in growth mode!...muscle is the longevity organ. the younger you are the better the start you have on longevity, your lifetime routines for longevity and healthy aging should be in a habit by about age 30. nobody wants sarcopenia!



Decline in skeletal muscle mitochondrial function with aging in humans



Abstract
Cumulative mtDNA damage occurs in aging animals, and mtDNA mutations are reported to accelerate aging in mice. We determined whether aging results in increased DNA oxidative damage and reduced mtDNA abundance and mitochondrial function in skeletal muscle of human subjects. Studies performed in 146 healthy men and women aged 18–89 yr demonstrated that mtDNA and mRNA abundance and mitochondrial ATP production all declined with advancing age. Abundance of mtDNA was positively related to mitochondrial ATP production rate, which in turn, was closely associated with aerobic capacity and glucose tolerance. The content of several mitochondrial proteins was reduced in older muscles, whereas the level of the oxidative DNA lesion, 8-oxo-deoxyguanosine, was increased, supporting the oxidative damage theory of aging. These results demonstrate that age-related muscle mitochondrial dysfunction is related to reduced mtDNA and muscle functional changes that are common in the elderly.
Many structural and functional changes occur with age in skeletal muscle in a wide range of species. In Caenorhabditis elegans, muscle changes resembling those in humans precede neuronal changes, and are a determinant of morbidity (1). Age-related muscle wasting, muscle weakness, and reduced aerobic capacity result in many metabolic disorders and diminished physical performance in humans (2–4). Reduced muscle mitochondrial function could contribute to age-related muscle dysfunction and reduced aerobic capacity. Increased prevalence of mtDNA mutations (5, 6) and decreased mtDNA abundance (7, 8) have been proposed as underlying causes of mitochondrial dysfunction in aging. This finding is based on a hypothesis that cumulative oxidative damage could be the cause of aging (9).

The rate of synthesis of contractile and mitochondrial proteins in human skeletal muscle declines with advancing age and may alter muscle metabolic capacity in older people (2–4). The activity of oxidative enzymes and content mRNA transcripts encoding mitochondrial proteins are also reduced in older muscles (3, 7, 10, 11). Reduced synthesis and activity of specific proteins can alter muscle functions. The major functional role of mitochondria is ATP generation, but it remains unclear whether mitochondrial ATP production rate (MAPR) in skeletal muscle declines with age in humans. Previous studies that attempted to address this question are not in agreement, reporting that MAPR is either unchanged with age (12–16) or declines (17–19). These differences may arise from the use in some studies of inadequate sample sizes, failure to account for wide variations in physical fitness and diet, and the inclusion of subjects with metabolic abnormalities or undergoing surgical procedures at the time of analysis. Most of the previous studies examined discrete groups of younger and older people so it is unclear whether changes in mitochondria occur continuously across the adult life span or arise more rapidly later in life. We therefore performed a comprehensive study to examine whether muscle mitochondrial function declines with age in humans by using a large group of well-characterized healthy men and women across a wide age span. We also sought to determine causes of age-related changes in mitochondrial function by examining the content of mitochondrial proteins, gene transcripts encoding mitochondrial proteins, mtDNA, and DNA oxidation.

snip

Results
Subject Characteristics. Body weight did not significantly change with age, but older people had reduced lean body and leg mass, and body fat and BMI were increased compared with young and middle-aged people (Table 1). Reported physical activity levels during the preceding year did not differ with age.


MAPR and Citrate Synthase Activity. MAPR per gram of muscle declined with age β‰ˆ8% per decade by using substrates that supply electrons primarily to complex I (e.g., glutamate plus malate) or complex II (succinate plus rotenone), respectively, of the respiratory chain (Fig. 1). Similar results were obtained for other substrates tested (not shown). After normalizing MAPR per milligram of mitochondrial protein, there was still a β‰ˆ5% decline per decade (Fig. 1).

Decline in muscle MAPR and citrate synthase activity with age. (A–C) MAPR is shown by using glutamate plus malate (A), pyruvate plus palmitoyl-Lcarnitine plus ketoglutarate plus malate (B), and succinate plus rotenone as substrates (C), respectively. (D) Citrate synthase activity. (E and F) MAPR by using glutamate plus malate and succinate plus rotenone, respectively, after normalization for mitochondrial protein. n = 146 for all measurements.
" data-icon-position="" data-hide-link-title="0" style="-webkit-font-smoothing: antialiased; box-sizing: border-box; background-color: transparent; font-weight: normal; text-decoration: none; outline: 0px !important; color: rgb(0, 90, 150); display: block; border: 0px; box-shadow: rgba(0, 0, 0, 0.15) 0px 2px 10px 0px;">Fig. 1.

Fig. 1.
Decline in muscle MAPR and citrate synthase activity with age. (A–C) MAPR is shown by using glutamate plus malate (A), pyruvate plus palmitoyl-Lcarnitine plus ketoglutarate plus malate (B), and succinate plus rotenone as substrates (C), respectively. (D) Citrate synthase activity. (E and F) MAPR by using glutamate plus malate and succinate plus rotenone, respectively, after normalization for mitochondrial protein. n = 146 for all measurements.

VO2max and Meal Glucose Tolerance. VO2max while cycling declined with age β‰ˆ8% per decade in both men and women (data not shown). VO2max was positively related to lean mass of the legs (r = 0.88, P < 0.001) and MAPR (r = 0.54, P < 0.001). After covariate adjustment for leg lean mass, which decreased 3% per decade (P < 0.01), the decline in VO2max was still 5% per decade (P < 0.001, Fig. 2). Furthermore, VO2max remained positively correlated with MAPR (Fig. 2). Together, leg lean mass and MAPR explained 86% of the variance in VO2max.

MAPR is related to VO2max and meal glucose tolerance. (A and B) VO2max, after covariate adjustment for leg lean mass, declined with age (A) and was positively associated with MAPR, n = 91 (B). (C) Fasting plasma glucose was not different between younger and older people (n = 10/group), but glucose excursion after a mixed meal was higher in older people (*, P < 0.05). (D) Post meal glucose area under the curve (AUC) was inversely related to MAPR.
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Discussion
The current study demonstrates that older people have significantly higher oxidative damage to DNA and that mtDNA abundance decreases with age. This decreased mtDNA abundance is associated with lower content of mRNA transcripts that encode mitochondrial proteins. We found that mitochondrial protein content and activity of a key oxidative enzyme (citrate synthase) are reduced in skeletal muscle from older people. There was also a continuous decline in mitochondrial capacity for oxidative phosphorylation (ATP production) with advancing age in skeletal muscle samples from a large number of healthy men and women between the ages of 18 and 87 years. The change in mitochondrial ATP production was closely related to VO2max and glucose tolerance after a mixed meal.

A major finding in the current study was that MAPR declined with age in a well characterized group of healthy adults when expressed either per unit of muscle mass or after normalization for mitochondrial protein. These results indicate that the decline in MAPR in older muscles is due to a combination of reduced mitochondrial content and a functional alteration in the existing mitochondrial population. An in vitro MAPR assay was used so it is possible that mitochondrial function could be affected during the preparation process. However, it is unlikely that this would cause a systematic age-related effect. Moreover, our results agree with recent findings that mitochondrial function is reduced in older people, assessed by either in vitro mitochondrial respiration (19) or in vivo NMR spectroscopy (17, 18). The present data were obtained from a larger number of people than used in the previous studies and there were no systematic differences in the quality of the mitochondrial preparations detected so the observed changes in MAPR appear to reflect a true change with age.

Many studies have shown that aerobic exercise enhances muscle mitochondrial biogenesis (25). We therefore controlled for physical activity by including only subjects who were not regularly performing vigorous physical activity. We also kept the participants on a standard diet for 3 days, confirmed that they were free of overt cardiovascular or metabolic disease, and were using few if any medications. However, as in nearly all studies of aging, it is impossible to exclude the possibility that there are undetected differences in daily physical activity between younger and older people that may be important, or that some of the observed changes could be due to an interaction between aging and sedentary behavior. Discrepancies among previous reports that addressed whether MAPR declines with age in human muscle may be related to the variable control over activity levels and diet. The inclusion of subjects who were undergoing limb surgery or suspected of having metabolic disorders (15, 16) may have also affected previous findings. Additionally, most investigations have been performed on either the quadriceps or calf muscles but the deltoid (15) or the tibialis anterior (12) were examined in two of the studies that reported no effect of age on MAPR, raising the possibility that age effects may vary among muscle groups. It was reported that the decline with age in citrate synthase activity differs between the vastus lateralis and gastrocnemius muscles (26), but additional comparisons among muscles in humans are needed to confirm this finding.

MAPR was closely associated with VO2max, even after adjusting for differences in leg lean mass. This finding suggests that muscle mitochondrial function is a determinant of VO2max in untrained individuals (27) and contributes to the decline in VO2max with advancing age. This finding is in contrast to exercise-trained individuals, in which VO2max is more limited by blood supply to the working muscle (28). In support of our findings, it was shown that oxygen uptake in contracting hindlimb muscles is reduced in old vs. young sedentary rats, even after matching for convective oxygen delivery, indicating that oxygen utilization in the mitochondria is reduced (29).

Another potential effect of mitochondrial dysfunction in older people is impaired glucose tolerance and diabetes (30, 31). Reduced MAPR in older people or first-degree relatives of people with type 2 diabetes was hypothesized to cause accumulation of intramuscular lipids and insulin resistance (17, 31). Consistent with that hypothesis, we found an inverse relationship between MAPR and the glycemic response to a mixed meal. However, glucose area under the curve was also positively related to trunk fat mass, measured by dual x-ray absorptiometry (r = 0.52, P < 0.01), and there is evidence that abdominal fat may be more important than either age or mitochondrial function for determining the increase in insulin resistance with aging (10, 32). Further, we recently showed that insulin stimulates transcription and translation of mitochondrial genes and proteins and increases MAPR, but this response is blunted in people with type 2 diabetes (21). Thus, we remain open to the possibility that insulin resistance contributes to mitochondrial dysfunction rather than the reverse. Further validation studies are therefore needed to establish the relationships between mitochondrial dysfunction, muscle lipid content, and insulin resistance.

We used a tandem MS method in conjunction with isotope labeling to quantify the content of multiple mitochondrial proteins in muscle samples from younger and older people. Previously, the activity of citrate synthase has been used as a measure of mitochondrial content (3, 10, 26). The decline in citrate synthase activity with age was supported by the proteomic analysis, in which citrate synthase and eight other mitochondrial proteins were reduced in older muscles. It should be noted that we focused our proteomic analysis on the 13 mitochondrial proteins that were present in all 10 pairs of samples from representative young and older people. Approximately 600 proteins have been identified in mitochondria from human heart muscle (33). However, the purpose of that investigation was to catalog as many proteins as possible in mitochondria isolated from several grams of tissue and the relative content of individual proteins was not reported. In contrast, our intent was to quantify multiple proteins from individual subjects. We therefore measured mitochondrial protein abundance within a whole-tissue homogenate to minimize the selective or variable loss of individual proteins during mitochondrial purification, and because using equal amounts of protein, as required for ICAT labeling, would minimize or eliminate the ability to detect the overall reduction in mitochondrial protein content. To our knowledge, this is the first demonstration of lower mitochondrial protein content in older human muscle by using this approach.

Content and function of specific proteins in muscle depends on protein synthesis and breakdown. Mitochondrial protein synthesis declines with age in human muscle (3). This decline may be due to reduced mRNA template availability because both COX3 and COX4 transcript levels declined significantly with age, in agreement with earlier work in rats and humans (7, 10, 11). Mitochondrial gene transcripts from both mtDNA and nuclear DNA are similarly reduced with age but it is not yet clear how coordinated expression between the genomes is controlled. We recently reported that mRNA abundance of three nuclear-derived transcription factors that regulate mitochondrial biogenesis [peroxisome-proliferator receptor coactivator 1 Ξ± (PGC-1Ξ±), nuclear respiratory factor 1 (NRF-1), and mitochondrial transcription factor A (TFAM)] did not change with age in human muscle (10). Thus, further work on the effect of age on the action of these and other nuclear signals that regulate mitochondrial biogenesis is needed.

Another important finding was that mtDNA content in muscle declined with age. This decline could contribute to mitochondrial dysfunction by reducing template availability for transcription and translation of key mitochondrial proteins. Collectively the present findings help explain the reduction in mitochondrial protein synthesis, protein content, activity of individual enzymes, and ultimately the ability of mitochondria to perform oxidative phosphorylation in older muscle.

The reduction in muscle mtDNA with aging could be attributable to accumulated oxidative damage (9). Our finding that DNA oxidation increased with age is consistent with this oxidative theory of aging and is in agreement with rodent studies that found higher DNA oxidation in older animals (34, 35). We used total DNA for this measurement to avoid introducing artifacts when separating nuclear and mtDNA, and therefore the measurement does not distinguish how the DNA damage is distributed in the nuclear and mitochondrial genomes. The level of oxidized bases is reported to be two to three times higher in mtDNA than nuclear DNA despite the fact that capacity to repair these lesions may actually be higher in mitochondria (36). The activity of antioxidant enzymes is also higher in older rat muscles (37). Together these results indicate that production rate of reactive oxygen species increases with age, exceeding the capacity of antioxidant defense enzymes and DNA repair. Oxidative damage is associated with increased mtDNA mutations and deletions in older muscles (5, 38). The importance of mtDNA damage was recently demonstrated in mice in which accumulation of mtDNA mutations resulted in accelerated aging and shorter lifespan (39). Oxidative damage to proteins, lipids, and other cellular components may also adversely affect the function of aging cells (40, 41).

In summary, the current study demonstrates that age-related reduction in muscle mtDNA and increased DNA oxidation is associated with reduced levels of mitochondrial gene transcripts and proteins. These changes are closely related to declining capacity for mitochondrial ATP production in skeletal muscle and collectively may contribute to lower physical function and higher insulin resistance that are common in older people.
 
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