AMEN-RA’S

AGE INHIBITION REGIMEN (AIR)

AN EXPOSITION ON THE EXPERIMENTAL, THEORETICAL & PRACTICAL

UNDERPINNINGS OF LIFESPAN EXTENSION
& OPTIMAL HEALTH PROMOTION,
A DISCOURSE ON THE METHOD OF GEROSTASIS

VOLUME II
Part II of IX

THE AMEN FAST

ABSTEMIOUS EATING ENABLES LIFE EXTENSION BY ENHANCING THE
INTEGRITY OF THE GENOME:
THE TALE OF TELOMERES
Dr. N!n Sava-Siva Amen-Ra, Dr.PH, MA, MSW, LGSW
 AMENTA ARCHIVES
© MMXV
Damascus, Maryland USA

ISBN 0-9741469-3-5

Correspondence concerning this publication is accepted at the following address:

Amen-Ra@AmentaEliteAthlete.com

Copies of this publication may be procured at the following website:

www.AmentaEliteAthlete.com

DISCLAIMER: This treatise does not constitute an attempt to dispense medical advice.
Rather, it is intended to inform individuals devoid of disease how they can best insure
freedom from infirmity and increase their odds of attaining exceptional longevity and vitality.
Information is offered and arguments are advanced in support of logically coherent claims
contained herein. Rational agents are hereby advised to rely principally on their own
reasoning faculties to critically evaluate the veracity of the Author’s statements. The Protocol
presented in the pages of this book is admittedly austere. As such, it will not appeal to the
masses of mankind nor ought it to be attempted by anyone deficient in discipline and
discernment—such efforts are bound to be abortive. There is risk inherent in anything
remotely rigorous and this Regimen is no exception to the rule. If, however, the attested
benefits of the Amen Protocol are substantive, the risk is arguably outweighed thereby.
 CONTENTS

LIFESPAN LENGTH & TELOMERE LENGTH: AVIAN EVIDENCE

LIFESPAN LENGTH & TELOMERE LENGTH: MAMMALIAN EVIDENCE

LIFESPAN LENGTH & TELOMERE LENGTH: AGGREGATE ‘ANIMALIAN’

EVIDENCE

LIFESPAN LENGTH & TELOMERE LENGTH: EPIDEMIOLOGIC EVIDENCE

LIFESPAN LENGTH & TELOMERE LENGTH: GUAGING GLYCEMIA

LIFESPAN LENGTH & TELOMERE LENGTH: GUAGING GLYCATION

CHOLESTEROL CURTAILS TELOMERES; FASTING SUPPRESSES

CHOLESTEROL SYNTHESIS: EXPLORING CONVOLUTED, YET COMPELLING
CORRELATIONS
 ______________________________________

“…[N]o physician, in so far as he is a physician, considers his own good in what he

prescribes, but the good of his patient; for the true physician is also a ruler having
the human body as a subject, and is not a mere money-maker….”

Plato, The Republic, Book I

_______________________________________

Proteins comprise the principal parts of the bodies of animals. Hence, sustaining the
structural soundness of our proteins should serve as the central strategy of any systematic
anti-aging agenda. This is certainly so with respect to our Regimen. However, it must be
understood that information is essential to the assemblage of proteins and such information is
embodied in the genome. As it were, the chemical constituents of the genome are
themselves subject to senescence. Consequently, any concerted effort to suppress senescence
must provide for the protection of the genome. Central to this provisionment of protection
are structures so important that the devotion of an entire installment of Volume II to their
discussion is indeed defensible. The Author has elected to organize our ensuing analysis in
terms of information acquired from particular classes of organisms—avian and mammalian
generally. Thence we turn to a truly integrative ‘Eco-Evolutionary’ analysis, summating the
science of senescence-stimulating telomere shortening in several species. Subsequently we
shall narrow our evidentiary analysis to epidemiological investigations encompassing that
most masterful member of the Order Primates, Man.

Lifespan Length & Telomere Length: Avian Evidence

Part II of the present Volume pertains primarily to a single molecular structure that is
essential to an understanding of aging and to interventions aimed at its inhibition: telomeres.
In Part III of the present Volume we shall see the importance of ensuring protein integrity in
our effort to attenuate the celerity of somatic senescence. Presently we wish to underscore
the essentiality of cellular integrity to our anti-aging agenda. Central to cellular integrity is the
maintenance of the replicative capacity of cells. Cells incur damage over time and must be
replaced in the interest of optimal physiological functioning. Cellular replacement is
dependent upon the duplication of chromosomes. Chromosomes cannot duplicate
indefinitely however. After a certain series of species-specific divisions, cellular senescence
ensues. At this point, proper cell processes are impaired and cellular suicide commonly
commences. The explanation for the inability of cells to divide incessantly centers upon the
progressive degradation of the ends of chromosomes with each successive cell division.
These ends of chromosomes, the Reader will recall from Part I, are called telomeres [Gk
!"#$% telos, end.] Though an extended excerpt from an article in The American Journal of
Human Biology was included in Part I of Volume II and contributed considerably to our
understanding of telomeres, we have elected to offer an additional introductory explanation
of this extremely important structure. As explained in the informative text, Human Molecular
Genetics:

[T]he ends of human chromosomes are protected by a repeat sequence

(TTAGGG)n, that is maintained by a special RNA containing enzyme
system, telomerase. Telomerase is present in the human germline but is absent
from most somatic tissues, and telomere length declines by 50-100 bp [base
pairs] with each cell generation. In prolonged culture, normal cells reach a
point of senescence, where they stop dividing.I

Since cellular senescence is associated with the truncation of telomeres, it seems reasonable
to expect that the biological age of organisms be likewise linked to telomere length. This
straightforward supposition was substantiated by a superb study published in the journal
Molecular Ecology. The Abstract explains that:

Explaining variation in life expectancy between individuals of the same age is

fundamental to our understanding of population ecology and life history
evolution. Variation in the length and rate of loss of the protective telomere
chromosome caps has been linked to cellular lifespan. Yet, the extent to
which telomere length and dynamics predict organismal lifespan in nature is
still contentious. Using longitudinal samples taken from a closed population
of Acrocephalus sechellensis (Seychelles warblers) [an avian (bird) species] studied
for over 20 years, we describe the first study into life-long telomere dynamics
(1-17 years) and their relationship to mortality under natural conditions
(n=204 individuals). We show that telomeres shorten with increasing age and
body mass, and that shorter telomeres and greater rates of telomere
shortening predicted future mortality. Our results provide the first clear and
unambiguous evidence of a relationship between telomere length and
mortality in the wild, and substantiate the prediction that telomere length and
shortening rate can act as an indicator of biological age [and] further to
chronological age when exploring life history questions in natural conditions.I

As to why body mass might contribute to a curtailment of telomere length we are left to
speculate. As long as we accept the inherent limitations of such speculation, the Author shall
advance a clarifying conjecture. Mass mirrors mitosis. That is, a multicellular organism’s
mass partly reflects the plentitude of its cells. Cellular plentitude is a product of cell division
(mitosis). Mitosis begins with chromosomal duplication. Duplication of chromosomes
catalyzes continued curtailment of telomere length. It should be considered that both fasting
and caloric restriction reduce the rapidity of cell replication and this anti-mitotic action may
mediate the life-extending effect of these interventions through a mechanism involving
telomere length modulation. This theoretical framework may link the most robust anti-aging
interventions (i.e. CR & CF) to a correspondingly robust index of aging (i.e. telomere
length).

IStrachan T & Read AP. Human Molecular Genetics, 3rd Edition, 2004: 499.
I
Barrett EL, Burke TA, Hammers M, Komdeur J, Richardson DS. Telomere length and dynamics predict
mortality in a wild longitudinal study. Molecular Ecology. 2013; 22(1):249-59.
 An analogous avian study published in the Proceedings of the National Academy of Sciences
found that finch lifespan likewise mirrors telomere length. The authors of the analysis
explained that:

The attrition of telomeres, the ends of eukaryotic chromosomes, is thought

to play an important role in deterioration with advancing age. The observed
variation in telomere length among individuals of the same age is therefore
thought to be related to variation in potential longevity. Studies of this
relationship are hampered by the time scale over which individuals need to
be followed, particularly in long-lived species where lifespan variation is
greatest. So far, data are based either on simple comparisons of telomere
length among different age classes or on individuals whose telomere length is
measured at most twice and whose subsequent survival is monitored for only
a short proportion of the typical lifespan. Both approaches are subject to
bias. Key studies, in which telomere length is tracked from early in life, and
actual lifespan recorded, have been lacking. We measured telomere length in
zebra finches (n=99) from the nestling stage and at the various points
thereafter, and recorded their natural lifespan (which varied from less than 1
to almost 9 y). We found telomere length at 25 d to be a very strong
predictor of realized lifespan (P<0.001); those individuals living longest had
relatively long telomeres at all points at which they were measured.
Reproduction increased adult telomere loss, but this effect appeared transient
and did not influence survival. Our results provide the strongest evidence
available of the relationship between telomere length and lifespan and
emphasize the importance of understanding factors that determine early life
telomere length. I

As with the previous investigation, the strength of this study lies in its longitudinal layout.
Both studies substantiate the supposition that the assessment of telomere length has
predictive power—more precisely, the power to predict avian lifespan. It is interesting to
note that reproduction was found to accelerate the attrition of telomeres. This makes sense
in the light of the Mass-Mitogen Model advanced by the Author above. Reproduction
augments elaboration of anabolic hormones. Anabolism entails increased mass and increased
mitosis. Increased mitosis promotes punctuation of telomere length. Testosterone,
androgens, estrogen and other anabolic steroidal hormones exert their mitogenic effects
especially (though not exclusively) upon organs of the reproductive system. Thus, the fact
that the discernible diminishment of telomere length linked to reproduction was insufficient
to ultimately impair longevity is not entirely surprising.

Evidence in avian organisms indicates that telomere length is predictive of lifespan.

I
Heidinger BJ, Blount JD, Boner W, Griffiths K, Metcalfe NB, Monaghan P. Telomere length in early life
predicts lifespan. Proceedings of the National Academy of Sciences (USA). 2012; 109(5):1743-8.
Lifespan Length & Telomere Length: Mammalian Evidence

The impressive investigations in avian organisms indicated above are reinforced by a

momentous metabolic study in a mammalian organism. Published in the journal Aging Cell,
this investigation endeavored to ascertain the effects of genetically altering the expression of
the enzyme telomerase on the physiological ages of mice of multiple strains. The Abstract is
instructive:

...We have determined the sera metabolite profile of 117 wild-type mice of
different genetic backgrounds ranging from 8 to 129 weeks of age. This has
allowed us to define a robust metabolomic signature and a derived
metabolomic score that reliably/accurately predicts the age of wild-type mice.
In the case of telomerase-deficient mice, which have a shortened lifespan,
their metabolomic score predicts older ages than expected. Conversely, in the
case of mice that overexpress telomerase, their metabolic score corresponded
to younger ages than expected. Importantly, telomerase reactivation late in
life by using a TERT-based gene therapy recently described by us
significantly reverted the metabolic profile of old mice to that of younger
mice, further confirming an anti-aging role for telomerase. Thus, the
metabolomic signature associated with natural mouse aging accurately
predicts aging produced by telomere shortening, suggesting that natural
mouse aging is in part produced by the presence of short telomeres. These
results indicate that the metabolomic signature is associated with the
biological age rather than with the chronological age. This constitutes one of
the first aging-associated metabolomic signatures in a mammalian organism.I

To clarify the importance of this study we should appreciate that it establishes (I) that
genetic diminishment of telomere length correlates with a metabolically senescent profile,
(II) that genetic enhancement of telomere length correlates with a metabolically juvenescent
profile and (III) that augmenting telomere length late in life through genetic manipulation
modifies the metabolic profile of mice, making the animal more juvenescent. It is this
potential to modify telomere length in mammals with the aim of suppressing senescence that
is of greatest interest to us.
A separate mammalian study has shown that the rapidity with which truncated
telomeres accumulate exerts an effect on lifespan. The investigators exposit thuswise:

Aberrantly short telomeres result in decreased longevity in both humans and

mice with defective telomere maintenance. Normal populations of humans
and mice present high interindividual variation in telomere length, but it is
unknown whether this is associated with their lifespan potential. To address
this issue, we performed a longitudinal telomere length study along the
lifespan of wild-type and transgenic telomerase reverse transcriptase mice.
We found that mouse telomeres shorten ~100 times faster than human
telomeres. Importantly, the rate of increase in the percentage of short
telomeres, rather than the rate of telomere shortening per month, was a

ITomas-Loba A, Bernardes de Jesus B, Mato JM, Blasco MA. A metabolic signature predicts age in mice. Aging
Cell. 2013; 12(1):93-101.
 significant predictor of lifespan in both mouse cohorts, and those individuals
who showed a higher rate of increase in the percentage of short telomeres
were also the ones with a shorter lifespan. These findings demonstrate that
short telomeres have a direct impact on longevity in mammals, and they
highlight the importance of performing longitudinal telomere studies to
predict longevity.II

The import of the above investigation lies in two principal findings. First, in keeping with
the Author’s Mass-Mitogen Model, rapid accumulation of short telomeres accelerates aging.
Short telomeres accumulate as a consequence of cell division, as a consequence of mitosis.
In this context, it would have been interesting to ascertain the existence of any correlation
between murine mass, the rate with which short telomeres accumulate, and eventual (i.e.
realized) lifespan. [Perhaps an experimentalist will be inclined to investigate such a
hypothesis issuing from the mind of a humble theorist such as the Author upon reading this
Treatise.] Second, the study comparatively quantifies the rate with which truncated telomeres
accumulate in the mouse and in Man, a rate that accords with the divergent lifespans of our
respective Families—Muridae and Hominidae.
The most momentous mammalian study to establish a relationship between
telomeres and longevity was undertaken by Spanish scientists from Madrid’s National
Cancer Centre. The paper was published in the Journal of the European Molecular Biology
Organization (EMBO). Before presenting the Abstract of the investigation in full, permit the
Author to clarify its content. The investigators inserted a gene governing the expression of
telomerase into an adenovirus—that is, into a viral vector. They then injected this viral
vector into two groups of experimental mice, aged 1 and 2 years respectively. They verified
the expression of the enzyme in the tissues of the telomerase-transfected mice. They
subsequently assessed various age-associated indices. Most importantly, they observed the
effect of the intervention on lifespan. As elaborated in the Abstract:

A major goal in aging research is to improve health during aging. In the case
of mice, genetic manipulations that shorten or lengthen telomeres result,
respectively, in decreased or increased longevity. Based on this, we have
tested the effects of a telomerase gene therapy in adult (1 year of age) and old
(2 years of age) mice. Treatment of 1- and 2-year old mice with an adeno-
associated virus (AAV) of wide tropism expressing mouse TERT
[telomerase] had remarkable beneficial effects on health and fitness, including
insulin sensitivity, osteoporosis, neuromuscular coordination and several
molecular biomarkers of aging. Importantly, telomerase-treated mice did not
develop more cancer than their control littermates, suggesting that the
known tumorigenic activity of telomerase is severely decreased when
expressed in adult or old organisms using AAV vectors. Finally, telomerase-
treated mice, both at 1-year and at 2-years of age, had an increase in median
lifespan of 24 and 13%, respectively. These beneficial effects were not
observed with a catalytically inactive TERT, demonstrating that they require
telomerase activity. Together, these results constitute a proof-of-principle of

IIVera E, Bernardes de Jesus B, Foronda M, Flores JM, Blasco MA. The rate of increase of short telomeres
predicts longevity in mammals. Cell Reports. 2012; 25; 2(4):732-7.
 a role of TERT in delaying physiological aging and extending longevity in
normal mice through a telomerase-based treatment, and demonstrate the
feasibility of anti-aging gene therapy.I

It is important to underscore the fact that overexpression of telomerase did not increase the
incidence of cancer. Among the alterations associated with carcinogenesis is the acquisition
of the ability to express telomerase, an ability that is absent or attenuated in most mature
cells (excepting sex cells or gametes). This ability permits cancer cells to divide incessantly,
the primary property responsible for their pathological character. Obviously, augmenting
expression of the enzyme would be otiose and inimical if cancer accrued along with lifespan
lengthening. Fortunately, this effect did not ensue. In a related observation, it is also
important to highlight the “healthspan” augmenting effects of the intervention—that is,
observed improvements in glycemia, physical fitness, coordination, and musculo-skeletal
integrity. Lengthening life without maintaining a high level of health would be less than
worthless.
We are, of course, more concerned with interventions that increase maximum lifespan
than those that merely increase average lifespan. Though the above abstract indicates only
the latter, the Results reveal the former. As stated therein:

To address whether our TERT [telomerase] gene therapy approach was able
to extend longevity, we performed Kaplan–Meier [statistical] analysis of mice
treated with either vectors [active or inactive for conferring telomerase
expression] in both age groups. We observed a significant extension of
lifespan in [active telomerase] TERT treated mice compared to the...non-
treated controls in both age groups. TERT treatment led to an increase in
survival of 24% in the 1 year-old group and of 13% in the 2 year-old group (p
< 0.05 for both groups) compared to...controls.... We also found a significant
increase in the median survival of the longest-lived mice (90th percentile) in
the [active telomerase] TERT group compared to the...controls, suggesting
that TERT may affect maximum longevity. Indeed, the longest-lived [active
telomerase] TERT treated mice from the 1 and 2 year-old groups surpassed
by 13 and 20%, respectively, the maximum longevity of the corresponding
longest-lived...controls. The positive effects on longevity may come from
both proliferating and non-proliferating tissues as [the viral vector] infects
both tissue types. Moreover, it has been recently demonstrated that
telomerase plays important roles in post-mitotic tissues.I

In short, maximum lifespan was increased by more than 10 percent in young adult mice and
20 percent in old adult mice.

IBernardes de Jesus B, Vera E, Schneeberger K, Tejera AM, Ayuso E, Bosch F, Blasco MA. Telomerase gene
therapy in adult and old mice delays aging and increases longevity without increasing cancer. EMBO (Molecular
Medicine). 2012; 4(8):691-704.
Figure II:II A. Kaplan-Mier curves illustrating extension of lifespan accompanying induced
expression of telomerase in mice. Excerpted from Bernardes de Jesus et alia 2012.
___________________________________

The above evidence persuades us that efforts to augment the expression of

telomerase or otherwise inhibit the attrition of telomeres with age may promote
prolongation of lifespan. This evidence becomes more persuasive when wedded with
evidence indicating that caloric restriction contributes to the lengthening and maintenance of
telomeres. Published in the Public Library of Science (PLOS), the study by Spanish investigators
Drs. Elsa Vera and colleagues employed animals whose expression of the enzyme telomerase
was experimentally increased. As explained in their Introduction:

...[W]e address here the effect of CR on telomere dynamics and telomere

function longitudinally during the lifetime of wild-type and telomerase
transgenic mice in a C57BL/6 genetic background, as well as study its impact
on several health indicators, cancer, and longevity. In this context, we
demonstrate that CR slows down telomere shortening and the accumulation
of telomere damage with aging in CR [wild-type] mice, a situation that
mimics mTERT over-expression. These positive effects of CR on telomere
length are observed in a wide range of tissues, including peripheral blood
mononuclear cells. Importantly, under our experimental settings TgTERT
mice under CR show a significant lifespan extension compared to wild-type
mice under CR. In contrast, wild-type mice under CR did not present a
significant lifespan extension compared to wild-type control mice. These
results demonstrate that CR synergizes with telomerase expression resulting
in a significant lifespan extension. A similar synergism was previously
observed between telomerase expression and higher level of tumor
suppressors, which result in a safe cancer protective background for
telomerase expression.I

Several aspects of the study summarized above warrant explication and elaboration. First,
the fact that caloric restriction was not found to extend lifespan in normal mice is plausibly
attributable to the fact that the control group was itself an experimental group. That is, the
control group was not permitted to feed freely in the customary ad libtum manner. Instead,
the control group was subjected to ~10% caloric restriction relative to a ‘true’ control group,
a group permitted to feed freely. Studies have shown (as cited and discussed in Volume I)
that even this moderate degree of dietary restriction can conduce to significant lifespan
extension. Moreover, as indicated also in Volume I, the condition of modern Man is
characterized by unfettered access to food—particularly palatable food. Thus, the most
meaningful ‘control’ group permitting extrapolation to humans is a true, ad libitum group.
Second, transgenic augmentation of telomerase, though it led to lifespan prolongation, also
induced an increased incidence of cancer. This effect mitigated the diminution of mortality
mediated by telomerase. Third, recognizing the cancer-thwarting effect of CR, the
investigators opined that combining telomerase treatment with CR might counter the
carcinogenic potential of transgenic telomerase, thereby revealing its otherwise salubrious,
anti-aging effect. This is precisely what their investigation established. Before we proceed, it
is prudent to reflect upon the capacity of transgenic telomerase induction to increase the
incidence of cancer. The Author cannot conscientiously condone a course of action whose
potential risks outweigh its potential benefits. In keeping with this commitment, I am
convinced by the available evidence that efforts to enhance the expression of telomerase (or
enhance the preservation of telomeres) is an important avenue of anti-aging science that
should be pursued when practicable. One such element of this evidence is an informative
investigation appearing in Oncogene. The summation of the study states that:

IVera E, Bernardes de Jesus B, Foronda M, Flores JM, Blasco MA. Telomerase reverse transcriptase synergizes
with calorie restriction to increase health span and extend mouse longevity. PLoS One. 2013; 8(1).
 Many degenerative diseases that occur with aging, as well as premature aging
syndromes, are characterized by presenting cells with critically short
telomeres. Telomerase reintroduction is envisioned as a putative therapy for
diseases characterized by telomere exhaustion. K5-mTert transgenic mice
overexpress telomerase in a wide spectrum of tissues. These mice have a
higher incidence of both induced and spontaneous tumors, resulting in
increased mortality during the first year of life. Here, we show that in spite of
this elevated tumor incidence and the initial lower survival, K5-mTert mice
show an extension of the maximum lifespan from 1.5 to 3 months,
depending on the transgenic line, which represents up to a 10% increase in
the mean lifespan compared to wild-type littermates. This longer lifespan is
coincidental with a lower incidence of certain age-related degenerative
diseases, mainly those related to kidney function and germline integrity.
Importantly, these effects of telomerase overexpression cannot be attributed
to dramatic differences in telomere length in aged K5-Tert mice compared to
wild-type mice....These findings indicate that telomerase overexpression
extends the maximum lifespan of mice.I

As to why augmented expression of telomerase might initially increase mortality in animal

models, it is possible that the inherently high rate of cell replication extant in younger
animals promotes the proliferation of already incipient cancer cells that would otherwise not
expand in the absence of inducement by telomerase. Telomerase can thus be viewed as a
permissive, rather than a causative agent in the context of cancer. However, when animals
attain adult dimensions and the rate of growth declines and desists, telomerase becomes
more benign, exerting a potent prolongevity effect. Following this reasoning, it seems
evident that children should not be exposed to agents that are known to appreciably
augment expression of telomerase. This amounts to a conservative caveat applicable to
innumerable bioactive agents that children should be advised to avoid.
We now return our attention to the investigation by Dr. Vera and colleagues. Most
important from the Amenite perspective is the finding that the combination of CR and
telomerase amplification lengthened lifespan beyond that of CR alone. This is evidence of
Selective Summation/Selective Synergism, our inveterate aim. As the Amenite Agenda
emphasizes information and interventions that can be practically implemented in the interest
of lifespan prolongation or gerostasis, what can we do with the evidence presented above?
Caloric restriction is already a cornerstone of our Regimen. Conversely, we cannot commit
ourselves to gene therapy; we won’t willingly inject genetically altered viruses into our bodies
bearing segments of telomeric DNA destined to integrate into our genomes, with the aim of
eliciting augmented expression of the life-extending enzyme telomerase. Even if this were
practicable, it would be problematic as this method of genetic modification can conduce to
carcinogenesis in some cases. Fortunately, as we shall soon see, there are viable alternative
adjuvants.

IGonzález-Suárez E, Geserick C, Flores JM, Blasco MA. Antagonistic effects of telomerase on cancer and aging
in K5-mTert transgenic mice. Oncogene. 2005; 24(13):2256-70.
 Evidence in mammalian organisms indicates that telomere length is predictive of
lifespan.

Lifespan Length & Telomere Length: Aggregate ‘Animalian’ Evidence

Epidemiological investigations ordinarily involve analyses of the human species solely.

However, this needn’t be strictly so. As the etymology of the appellation implies, it
essentially entails any intellectual labor, undertaking, or insight (hence, logos, logia, logical)
centered upon (hence, epi) entire or particular segments of populations (hence, demos). And
though only exceptional orders of animals deliberately design what can be deemed a demos,
domus, or discernible domiciles in the manner of Man, they can correctly be conceived as
constituting population clusters. More momentous and interesting than the Author’s
elaborate etymological excursion is the applicability and utility of epidemiological tools or
techniques, the ability of epidemiology to elucidate biological phenomena, especially
aggregately. The two scientists, the results of whose research shall occupy this single section
of our Treatise, exquisitely illustrate what we might call ‘Animalian Epidemiology’ in action.
For they have succeeded in synthesizing the research we recounted above into an organized
framework, clustering classes and species and quantifying correlations between telomere
length and lifespan thereamong. Their Abstract in Experimental Gerontology informs us that:

Research on the physiological causes of senescence aim to identify common

physiological mechanisms that explain age-related declines in fitness across
taxonomic groups. Telomeres are repetitive nucleotide sequences found on
the ends of eukaryotic chromosomes. Past research indicates that telomere
attrition is strongly correlated with inter-specific rates of aging, though these
studies cannot distinguish whether telomere attrition is a cause or
consequence of the aging process. We extend previous research on this topic
by incorporating recent studies to test the hypothesis that telomeres shorten
more slowly with age in slow-aging animals than in fast-aging ones. We
assembled all studies that have quantified cross-sectional (i.e. between-
individual) telomere rates of change (TROC) over the lifespans of wild
animals. This included 22 estimates reflecting absolute TROC (TROCabs,
bp/yr., primarily measured using the terminal restriction fragment length
method), and 10 estimates reflecting relative TROC (TROCrel, relative
telomere length/yr., measured using qPCR), from five classes (Aves,
Mammalia, Bivalvia, Reptilia, and Actinopterygii). In 14 bird species, we
correlated between-individual (i.e. cross-sectional) TROCabs estimates with
both maximum lifespan and a phylogenetically-corrected principle
component axis (pcPC1) that reflected the slow-fast axis of life-history
variation. Bird species characterized by faster life-histories and shorter
maximum lifespans had faster TROCabs. In nine studies, both between-
individual and within-individual TROC estimates were available (n=8 for
TROCabs, n=1 for TROCrel). Within-individual TROC estimates were
generally greater than between-individual TROC estimates, which is
indicative of selective disappearance of individuals with shorter telomeres.
However, the difference between within- and between-individual TROC
 estimates was only significant in two out of nine studies. The relationship
between within-individual TROCabs and maximum lifespan did not differ
from the relationship of between-individual TROCabs and maximum
lifespan. Overall, our results provide additional support for the hypothesis
that TROC is correlated with inter-specific rates of aging and compliment
the intra-specific research that also find relationships between telomere
attrition and components of fitness.I

Refreshingly, the rarity of this research is reflected in the rationality of the researchers
themselves. Permit me to expound Amenites. Often the Author is impelled to underscore
the implications of investigations undertaken by scientists who seem insufficiently informed
about the broader context of their area of investigation and often oblivious to the practical
consequences of their findings—this despite promising pronouncements pertaining to
translational biomedical research; meaning research able to be applied rapidly and realistically
to the augmentation of human health, vitality, and longevity. In contrast to some exceedingly
cloistered scientists, Drs. Dantzer and Fletcher perceive and propound the significance of
their entirely non-experimental study: Both within and between diverse species, short
telomeres signify shorter lifespans whereas longer telomeres are linked to longevity. Simple
and (so far seemingly) unambiguous. This insightful epidemiological analysis aimed at the
Animal Kingdom has its counterparts in Man. In the ensuing section, we shall see these
conceptual counterparts.

Short telomeres equate with truncated lifespan; long telomeres equate with lengthened
lifespan. This is what the aggregate evidence in animals indicates.

Lifespan Length & Telomere Length: Epidemiologic Evidence

The mammal we are most interested in is Man. It is necessary to note that mice are more
prolific in their production of paltry telomeres than humans, whose rate of aging is ~25
times slower than that of murids. Importantly, epidemiological investigations have
convincingly confirmed that telomere length is a predictor of life expectancy in our species.
One such analysis appeared in the journal Aging Cell. It is this study that we shall focus our
attention upon in consideration of its clarity, its conclusions, and its comportment with a
general theoretical framework formerly fashioned by the Author:

Leukocyte telomere length (LTL) is ostensibly a bio-indicator of human

aging. Here we report that African Americans have longer LTL than whites.
We studied cross-sectionally 2453 individuals from the National Heart, Lung,
and Blood Study (age = 19-37 years), comprising 1742 whites and 711
African Americans. We measured LTL by Southern blots of the terminal
restriction fragments length. In 234 participants, telomere repeats were also
measured by quantitative polymerase chain reaction (qPCR). Adjusted for age
and body mass index (BMI), the respective leukocyte telomere lengths (mean
+/- SEM) were considerably longer in African Americans than in whites

IDantzer B, Fletcher QE. Telomeres shorten more slowly in slow-aging wild animals than in fast-aging ones.
Experimental Gerontology. 2015 Sep 5. pii: S0531-5565(15)30036-X.
 both in the Family Heart Study (7.004 +/- 0.033 kb [kilobases, i.e. units of
one thousand nucleotide bases of DNA] vs. 6.735 +/- 0.024 kb, p<0.0001).
We confirmed the racial effect on LTL by qPCR (3.038 +/- 0.565 T/S units
for African Americans vs. 2.714 +/- 0.487 T/S units for whites, p<0.001).
Cross-sectionally, sex- and BMI-adjusted LTL became shorter with age
(range 19-93 years) at a steeper slope in African Americans than in whites
(0.029 kb/year) vs 0.02 kb/year), respectively, p=0.0001). We suggest that
racial difference in LTL arises from a host of interacting biological factors,
including replication rates of hematopoietic stem cells.I

First, it is important to appreciate that the investigators confirmed a consistent finding—that

telomeres shorten as organisms age, the organism in this case being Man. [More specifically,
the scientists strengthened the association between age and telomere attrition. Because the
study was cross-sectional instead of longitudinal it is not definitively confirmative of
senescence-associated shortening of telomeres as a properly prospective project would
prove.] Now we note the novel knowledge. African Americans exhibited longer telomeres
than Caucasian Americans. ‘Of what significance is this?’ the thoughtful Amenite or other
Intellect might inquire. Recall the reason for our reliance upon animal evidence in the arena
of Biogerontology. Simply stated, humans live too long to permit protracted interventional
studies, the termini of which are differential death. We therefore avail ourselves of animal
studies wherein the effects of interventions (e.g. caloric restriction, fasting, neutraceutical or
pharmaceutical agents, genetic modification, &c.) can be observed within a practical period.
However, if a robust marker of aging has been identified in diverse organisms, and this
indicator is indeed applicable to Man, analyzing differences in such an aging index can yield
important insights into the nature of senescence in our own species. This is evidently the
case with telomeres. As explained by the authors in their informative Introduction:

Mounting evidence suggests that leukocyte telomere length (LTL) is a bio-

indicator of human aging, cardiovascular aging in particular. LTL is heritable,
although it is unknown how much of this heritability relates to birth LTL and
the rate of its shortening from birth onward. Age-dependent LTL shortening
is due to successive divisions of hematopoietic stem cells (HSCs) and
progenitor cells (PCs) that form peripheral leukocytes. Inflammation and
oxidative stress—two central elements in the biology of aging and aging-
related diseases—were reported to be associated with LTL. Inflammation
entails an increase in number and diminished biological life of leukocyte
subsets in the circulation, which would heighten the demand on HSCs/PCs
to replicate, a phenomenon expressed in an accelerated telomere attrition and
ultimately shortened LTL. Oxidative stress heightens the loss of telomere
repeats per cell division. The compounded effect of inflammation/oxidative
stress on the paces of both aging and LTL attrition conceivably explains the
shortened LTL observed in individuals with aging-related diseases,

IHunt SC, Chen W, Gardner JP, Kimura M, Srinivasan SR, Eckfeldt JH, Berenson GS, Aviv A. Leukocyte
telomeres are longer in African Americans than in whites: the National Heart, Lung, and Blood Institute Family
Heart Study and the Bogalusa Heart Study. Aging Cell. 2008; 7(4):448-450.
 particularly atherosclerotic cardiovascular (CV) disease, which is strongly
linked to inflammation.I

Among other things, the preceding excerpt sheds additional light on the interrelationship
between aging, adiposity, and inflammation. Adiposity increases inflammation owing to the
exudation of inflammatory adipokines by overly abundant adipose cells. Excessive
inflammation amplifies replication of white blood cells (i.e. leukocytes), accordingly
accelerating telomere attrition. Telomere attrition, in turn, catalyzes aging. We shall return to
this topic in the ensuing section.
The significance of the study by Dr. Steven Hunt and his colleagues lies partly in its
pedagogic parsimony. An explanation is now in order. It was in Evolutionary Nutrition that the
Author initially advanced his meta-theory of human evolution. Central to this theory is the
postulation that humans harbor adaptations to an austere evolutionary environment, the
climatic conditions of which catalyzed chronic caloric restriction among the ancestors of
Man. Originally, this ancestral population was entirely Africoid. Among the adaptations
incurred by our Africoid ancestors was increased metabolic efficiency—that is, the ability to
subsist on a paltry portion of provisions. We are aware of the broad, biological relationship
between metabolic rate and interspecific longevity—animals with longer lifespans have lower
rates of energy expenditure. Europoid peoples are an evolutionary emanation or off-shoot of
the ancestral Africoid population. Among the reasons for racial differentiation was the
nature of the environment to which the ancestors of Europoids migrated and adapted—a
cold climate in which heightened energy expenditure and energy intake were equally
essential, equally advantageous. A consistent consequence of heightened energy intake and
expenditure is accelerated aging or reduced lifespan potential. Under the auspices of
undertaking his doctoral degree in Epidemiology, the Author confirmed the hypothesis that
Africoid peoples (specifically African Americans) exhibit lower metabolic rates than
Europoid peoples (specifically Caucasian Americans).II It was the Author’s intention to
combine this component of his Dissertation with a comparative analysis of maximum
lifespan potential in African Americans relative to Caucasian Americans. Unfortunately,
unavoidable academic and administrative impediments precluded the pursuit of this project.
Other, independent sources of information indicate that the Author’s theoretical
framework—particularly the postulation of heightened lifespan potential in persons of
appreciable Africoid admixture—is scientifically sound. One such source of information is
the body of evidence in question. In establishing that African Americans exhibit appreciably
longer telomeres than European Americans, researchers have reinforced the foundation of
the Author’s Hypometabolic Theory of Human Evolution, albeit unbeknownst thereto.
Preliminarily, it is important to understand that the instantaneous essence of cross-sectional
studies commonly permits comparisons between groups. Obversely, the extended essence of
longitudinal studies commonly permits comparisons within groups, as the individuals
included therein can serve as their own internal controls from one time-point to another.
Principally, this is what makes the prospective study performed by Dr. Rewak of Harvard’s
School of Public Health particularly powerful, poignant, and persuasive. For she and her

IHunt SC, Chen W, Gardner JP, Kimura M, Srinivasan SR, Eckfeldt JH, Berenson GS, Aviv A. Leukocyte
telomeres are longer in African Americans than in whites: the National Heart, Lung, and Blood Institute Family
Heart Study and the Bogalusa Heart Study. Aging Cell. 2008; 7(4):448-450.
IIAmen-Ra N, Velasco-Mondragon E, Hossain M, Bronner Y. Energy expenditure differs between Black and

White Americans: implications for obesity prevention research. Food & Nutrition Sciences. 2012; 3(7):914-924.
team tested telomere length both at birth and later in time in a longitudinal fashion. This is
how they summarized their findings in the journal Biological Psychology:

Recent work suggests that leukocyte telomere length (LTL), a marker of

cellular aging, is sensitive to effects of social stress and may also provide early
indication of premature aging. Using data from a birth cohort with LTL
information at birth and in middle adulthood we examined a potential source
of race-based health disparity by testing the hypothesis that Blacks would
demonstrate a faster rate of telomere shortening than Whites. Linear
regression analyses were conducted and adjusted for pack years, BMI,
education and social factors, diet, exercise, marital status, and age. At birth
Black individuals had LTLs that were longer, on average, than their White
counterparts (&=3.85, p<0.01). However, rate of shortening was greater for
Blacks, who showed a larger difference in length between birth and
adulthood (&=5.10, p=0.01) as compared with Whites, resulting in smaller
racial differences in absolute adult LTL.I

What does this study signify? Several things, though we shall focus on only a few. First, it
empirically establishes what the Author’s evolutionary hypothesis postulated a decade
prior—that Africoid peoples possess a longer lifespan potential, evidenced in this particular
case by longer telomeres at birth. It also affirms the Author’s explanation, offered in
Evolutionary Nutrition, that African Americans tend to attain lower average lifespan owing to
adverse environmental factors that are largely self-imposed. Such factors frequently found
among Blacks are basically behavioral in nature, encompassing overindulgence, inadequate
activity, and poor diet quality coincident with a phenotype fashioned for metabolic efficiency
and a paucity of proteinaceous products, particularly meat.

It is important to inform the Reader that the phenomenon of “mortality crossover”

is generally acknowledged in epidemiological, gerontological, and actuarial circles—that is,
the observation that the African American population exhibits a consistent upward shift in
survival at the extremity of the aging curve (especially at the nonagenarian and centenarian
juncture). Arguably, this is indicative of an inherent, innate, adaptive attribute enabling
attainment of advanced age among African Americans. The Author’s identification of
significant, population-level differences in metabolism between Blacks and Whites
substantially solidifies this theoretical framework. So too does Dr. Hunt’s finding that
African Americans harbor lengthier telomeres and Dr. Rewak’s ascertainment of the fact
that African Americans’ telomeres are indeed lengthier than their congenitally less longevous
leucous brethren even at birth. Of interest in this respect is Dr. Hunt’s sensible speculation
that the observed differences in telomere length between Blacks and Whites may be
attributable to differential rates of cell replication. Importantly, reduced cellular replication is
reflective of a reduced rate of metabolism, the very phenomenon that the Author’s study
established among African Americans. Hence, despite the fact that definitive, experimental
evidence is all-but impossible to obtain in the area of human Biogerontology, the

IRewak M, Buka S, Prescott J, De Vivo I, Loucks EB, Kawachi I, Non AL, Kubzansky LD. Race-related health
disparities and biological aging: does rate of telomere shortening differ across blacks and whites? Biological
Psychology. 2014; 99:92-9.
accumulation of evidence from independent sources increases our confidence that integral
aspects of our avowed understanding of human aging are indeed accurate.

Epidemiological evidence accords with experimental and observational animal

evidence indicating that aging is associated with attrition of telomeres. Inherent differences
in the rate of telomere truncation as evidenced by longer telomeres in Africoid peoples may
underlie observed differences in lifespan potential between races.

Lifespan Length & Telomere Length: Gauging Glycemia

We are aware that a fundamental effect of fasting is the suppression of serum glucose
concentration. We are also aware that lifelong, cyclic fasting is sufficient to induce extension
of animal lifespan. We shall now explore evidence linking these two observations. The link
lies largely in the length of telomeres and the effect of sugar in shortening them. This basic
physiological finding was discerned by Dr. Dabouras and colleagues and published in the
journal Neuroscience Letters. The exposition of their investigation is technical yet telling. They
employed an in vitro cellular system utilizing neurologic cells called oligodendrocytes. An
excerpt from their Abstract ensues:

To investigate aspects of aging on rat oligodendrocytes, cells of an

oligodendrocyte cell line, so-called OLN-93, were cultured either in the
presence or absence of glucose. Our data demonstrated that glucose-induced
aging in vitro caused an elongation and thickening of cell processes and
significantly increased the expression of netrin [a receptor mediating multiple
developmental functions including positional guidance of neuronal growth]
reflecting a more mature state of oligodendrocyte development. A possible
age-inducing effect of glucose is also supported by...shortening of telomeres
in glucose-treated oligodendrocytes.I

Despite its deceptively daunting terminology, the study is simple and substantive in its
significance. It straightforwardly shows that sugar somehow shortens the telomeres of a
subset of cells specific to the central nervous system. As the investigators intimate, this effect
is evidently an accompaniment of aging at the cellular level. Does dextrose directly diminish
telomere length? The answer to this question, as a study by Japanese investigators from
Nagoya University’s Department of Geriatrics seems to suggest, is not necessarily. Rather, it
seems that sugar impairs the enzyme that enables the maintenance of telomeres—that is,
high glucose concentration compromises the catalytic activity of the enzyme telomerase. As
explained in their abstract:

The elderly are prone to postprandial hyperglcemia that increases their

cardiovascular risk. Although insulin therapy is necessary to treat diabetes,
high plasma concentrations of insulin may cause the development of

IDabouras V, Rothermel A, Reininger-Mack A, Wien SL, Layer PG, Robitzki AA. Exogenous application of
glucose induces aging in rat cerebral oligodendrocytes as revealed by alteration in telomere length. Neuroscience
Letters. 2004; 368(1):68-72.
 atherosclerosis and accelerate endothelial senescence. We assumed that high
glucose causes stress-induced premature senescence and replicative
senescence and examined the regulatory role of insulin in endothelial
senescence and functions under different glucose conditions. Exposure of
human endothelial cells to high glucose (22 mM) for 3 days increased
senescence-associated-&-galactosidase activity, a senescence marker, and
decreased telomerase activity, a replicative senescence marker. Physiological
concentrations of insulin preserved telomere length and delayed endothelial
senescence under high-glucose conditions. The effect of insulin under high
glucose conditions was associated with reduced reactive oxygen species and
increased nitric oxide (NO)….I

In the in vitro system employed by these scientists, exposure of human cells to heightened
sugar suppressed the activity of telomerase and significantly shortened the ends of
chromosomes. As expressed in their Results section:

Telomerase activity decreased significantly after 3 days of exposure to high

glucose in [human cell culture], and subsequently, telomere length was
significantly shortened by 4 weeks, which indicated replicative senescence.I

To reiterate, this finding affords us increased insight into the mechanism by which elevated
glucose exerts a deleterious effect on telomeres as a consequence of enzyme inhibition. Is
such sugar-mediated telomere-truncation as established by these two studies evident also at
the organismal level? Stated alternatively, is there an in vivo analogue to the aforecited in vitro
studies? This is what Indian investigator Dr. Monickaraj and colleagues found. More
specifically, they sought to determine whether diabetes (i.e. decidedly ‘dietetically-
determined’, Type II diabetes mellitus) is associated with telomere truncation. The rationale
for this approach is clear—diabetics constitute a clinical cohort characterized by elevated
circulating sugar; ascertaining any difference in telomere length between diabetics and
persons devoid of diabetes strongly implicates elevated sugar as a likely etiological agent in
the shortening of telomeres. Implicated it is. As the Abstract explains:

Although shortened telomeres were shown to be associated with several risk

factors of diabetes, there is lack of data on their relationship with
mitochondrial dysfunction. Therefore, we compared the relationship
between telomere length and mitochondrial DNA (mtDNA) content in
patients with type 2 diabetes mellitus (T2DM; n = 145) and in subjects with
normal glucose tolerance (NGT; n =145). Subjects were randomly recruited
from the Chennai Urban Rural Epidemiology Study. mtDNA content and
telomere length were assessed by Real-Time PCR. Malonodialdehyde, a
marker of lipid peroxidation was measured by thiobarbituric acid reactive
substances (TBARS) using fluorescence methodology. Adiponectin levels

IMatsui-Hirai H, Hayashi T, Yamamoto S, Ina K, Maeda M, Kotani H, Iguchi A, Ignarro LJ, Hattori Y. Dose-
dependent modulatory effects of insulin on glucose-induced endothelial senescence in vitro and in vivo: a
relationship between telomeres and nitric oxide. The Journal of Pharmacology & Experimental Therapeutics. 2011;
337(3): 591-9.
 were measured by radioimmunoassay. Oxidative stress as determined by lipid
peroxidation (TBARS) was significantly (p < 0.001) higher in patients with
T2DM compared to NGT subjects. In contrast, the mean telomere length,
adiponectin and mtDNA content were significantly (p < 0.001) lower in
patients with T2DM compared to NGT subjects. Telomere length was
positively correlated with adiponectin, HDL, mtDNA content and good
glycemic/lipid control and negatively correlated with adiposity and insulin
resistance. On regression analysis, shortened telomeres showed significant
association with T2DM even after adjusting for waist circumference and
adiponectin but lost its significance when further adjusted for telomere
length, TBARS and insulin resistance. Our study emphasizes the clustering of
accelerated aging features viz., shortened telomeres, decreased mtDNA
content, hypoadiponectinemia, low HDL, and increased oxidative stress in
Asian Indian type 2 diabetes patients.I

Ignoring for our present purposes the investigators’ interest in such extraneous outcomes as
mitochondrial DNA content, oxidative stress, and adiposity, we focus instead on the most
pertinent finding: Diabetes (and, presumably, heightened serum glucose concentration)
conduces to the curtailment of telomeres. The Reader is herein reminded that diabetes is
associated with physiological phenomena indicative of accelerated aging. Thus, the
association between diabetes and telomere length logically links both states to senescence.
From our perspective, a study conducted by Dr. Ahmad and colleagues and
published in the journal Diabetic Medicine is somewhat superior to the aforecited investigation.
It is superior insofar as it involved individuals devoid of diabetes and, unlike Dr. Monickarj’s
study, it analyzed actual glucose concentration and correlated this measure with telomere
length. [Recall that the Amen Protocol is principally directed at individuals devoid of
discernible disease]. In their informative Abstract, the Swedish investigators from the
Genetic and Molecular Epidemiology Unit of Lund University summarized their study and
explained its underlying logic:

Background:...Skeletal muscle is a major metabolic organ and plays important

roles in glucose metabolism, insulin sensitivity, and insulin action. Muscle
telomere length reflects the myocyte’s exposure to harmful environmental
factors. Leukocyte telomere length is considered a marker of muscle telomere
length and is used in epidemiological studies to assess associations with
ageing-related diseases where muscle physiology is important. However, the
extent to which leukocyte and muscle telomere lengths are correlated is
unknown, as are their relative correlations with glucose and insulin
concentrations. The purpose of this study was to determine the extent of
these relationships.

Methods: Leucocyte and muscle telomere length were measured by

quantitative real-time polymerase chain reaction in participants from the
Malmö Exercise Intervention (n=27) and the Prevalence, Prediction and

IMonickaraj F, Aravind S, Gokulakrishnan K, Sathishkumar C, Prabu P, Prabu D, Mohan V, Balasubramanyam

M. Accelerated aging as evidenced by increased telomere shortening and mitochondrial DNA depletion in
patients with type 2 diabetes. Molecular & Cellular Biochemistry. 2012; 365(1-2):343-50.
 Prevention studies (n=31). Participants in both studies were free from Type
2 diabetes. We assessed the association between leucocyte telomere length,
muscle telomere length and metabolic traits using Spearman correlations [a
statistical technique enabling the determination of the degree of association
between variables] and multivariate linear regression. [Statistical] analysis was
used to assess agreement between leucocyte and muscle telomere length.

Results: In age-, study-, diabetes family history- and sex-adjusted models,

leucocyte and muscle telomere length were positive correlated (r = 0.39, 95%
CI 0.15-0.59). Leucocyte telomere length was inversely associated with 2-hr
glucose concentrations (r = -0.58, 95% CI -1.0 to -0.16), but there was no
correlation between muscle telomere length and 2-h glucose concentration
(r=0.05, 95% CI -0.35 to 0.46) or between leucocyte or muscle telomere
length with other metabolic traits.

Conclusions: In summary, the current study supports the use of leucocyte

telomere length as a proxy for muscle telomere length in epidemiological
studies of Type 2 diabetes aetiology.I

As is commonly the case, our unbeknowing empirical emissaries were interested in ideas
extraneous to the Amenite Agenda. These we can appropriately ignore. The diligent
scientists have, however, done us a definite service in determining that the lower the level of
circulating sugar, the longer the length of telomeres in the leucocytes of the subjects.
Additionally, it was rather astute of these investigators to ascertain whether the length of
telomeres in the genetic material of the subjects’ muscles similarly correlated with the
concentration of sugar. It did not, as the careful Reader will have noted. However, Amenites
are inclined to imagine that a significant association may have been observed if the glucose
concentration had been assessed after a lengthier fast—if not of an elongated Amenesque
interval of 22-23 hours, then assuredly more than the 2 hour interval adopted by the
investigators. In defense of my epidemiological colleagues, however, it is not easy to impose
idealistic observational or experimental conditions on animals of such a ‘sapient’ species as
Man. Instead, we must often “get in where we fit in”. Skeletal muscle is less labile than
leukocyte cells, renewing their constituents far less rapidly. Consequently, the length of
telomeres in muscle may reflect fasting glucose concentration over a more considerable
temporal interval. A study confirming (or disconfirming) the Author’s speculative contention
could be carried out in an animal model with relative ease. This illustrates the important idea
that, although we are interested in the attenuation of aging in humans, animal studies are
often far superior in terms of the scientific information they are able to afford us. This is
precisely why our multivolume work relies so substantially on evidence acquired from
studies in nonhuman animals, and often accords clinical and epidemiological investigations a
secondary scientific status.

IAhmad S, Heraclides A, Sun Q, Elgzyri T, Ronn T, Ling C, Isomaa B, Eriksson KF, Groop L, Franks PW,
Hansson O. Telomere length in blood and skeletal muscle in relation to measures of glycaemia and
insulinaemia. Diabetic Medicine. 2012; 29(10):e377-81.
 A most interesting analysis was undertaken by Spanish investigators from the
University of Barcelona’s Neuroscience Institute. Published in the journal Brain, Behavior, and
Immunology, the principal purport of the study was to ascertain the association among glucose
regulation, telomere length, and depression. As a means of ascertaining glycemic control, the
investigators employed a 2-hour glucose tolerance test. This test involves administering a
sugar solution to subjects and gauging the concentration of serum glucose after an interval
of 2 hours. Essentially, what they found was that depressed subjects exhibited elevated ‘post-
load’ glucose concentration and, importantly, shorter leukocyte telomere length. As their
informative abstract explains:

Chronic mood disorders have been associated with a shortened telomere, a

marker of increased mortality rate and aging, and impaired cellular immunity.
However, treatment may confound these relationships. We examined the
relationship of glucose tolerance, white blood cell count and telomere length
to depression in newly diagnosed, anti-depressant-naïve patients. Subjects
with major depression (n=15), and matched healthy control subjects (n=70)
underwent a two-hour glucose tolerance test and evaluation of blood cell
count and telomere content. The depression group had significantly higher
two-hour glucose concentrations and a lower lymphocyte count than control
subjects (respective means [SD] for two-hour glucose were 125.0 mg/dL
[67.9] vs 84.6 [25.6] (p<.001)….Telomere content was significantly shortened
in the depression group (87.9 [7.6]) compared to control subjects (101.0
[14.3]; p<0.01). Abnormal glucose tolerance…and a shortened telomere are
present in the course of depression independently of the confounding effect
of antidepressant treatment, supporting the concept of major depression as
an accelerated aging disease.I

This study is of interest in view of its validation of the link between elevated glucose
concentration and corresponding curtailment of telomere length. It is also of interest in that
it underscores an underappreciated relationship between glycemia, mental illness, mortality,
and aging. Even if the relationship between hyperglycemia and depression is not causal, it is
clear that the mitigation of hyperglycemia can counter the curtailment of telomere length.
Among individuals inclined to develop Depression, an Amenesque intervention aimed at
attenuating glycemica may mitigate the increased risk of mortality to which they are subject,
owing to the telomeric protection proffered by our Protocol. There is even the plausible
possibility that our System, in aggregate, may diminish Depression in a deeper, more
fundamental way by modulating the molecular mediators of this most prevalent mental
disorder.

Excursus: The Author is presently undertaking research entailing the application of

the AOHP to the area of addiction. Denominated the Amen Anti-Addiction
Intervention (AAI), it offers the prospect of preventing and treating diverse
addictions through a single, multi-modal intervention encompassing fasting,
scheduled feeding, caloric restriction, rigorous exercise, selective supplementation,

IGarcia-Rizo C, Fernandez-Egea E, Miller BJ, Oliveira C, Justicia A, Griffith JK, Heaphy CM, Bernardo M,
Kirkpatrick B. Abnormal glucose tolerance, white blood count, and telomere length in newly diagnosed,
antidepressant-naive patients with depression. Brain, Behavior, and Immunology. 2013; 28:49-53.
 and meditation. The Author’s preliminary investigations indicate that each of the
aforementioned aspects of the Intervention alter fundamental physical and
behavioral factors that influence addictive behaviors. Importantly, Depression is a
disorder that is intimately associated with addiction. In light of the above finding
that Depression is associated with heightened glucose concentration and impaired
glucose regulation and given the fact that the Amen Protocol is so effective in
attenuating hyperglycemia/dysglycemia, it is clear that this aspect of our Regimen is
yet another means by which addictive (and depressive) symptomatology can be
suppressed.

In Part I of Volume II, we encountered an epidemiological investigation by Dr.

Miryoung Lee and colleagues. Though the researchers’ principal interest lay in
cardiovascular-related outcomes or the interrelationship between adiposity, anthropometric
measures, and telomere length, they also, importantly, ascertained an inverse relationship
between fasting glucose concentration and telomere length. That is, they found that among a
cohort of “normoglycemic” individuals, the lower the fasting glucose concentration, the
lengthier the participants’ telomeres. As the Abstract delineates, they sought to:

Background:...[A]ssess the relationship between telomere length and adiposity,

using dual-energy X-ray absorptiometry (DXA) and magnetic resonance
imaging (MRI), in addition to conventional anthropometric proxies including
body mass index (BMI) and cardiovascular disease risk factors.

Methods: A cross-sectional sample of 309 non-Hispanic white participants in

the Fels longitudinal Study aged 8 to 80 yr (52% female) was included.
Average telomere length was measured by quantitative PCR [i.e. polymerase
chain reaction].

Results: Telomere length was negatively correlated with age (r = -0.32,

P<0.0001) and had numerous significant correlations with established
cardiovascular disease risk factors including waist circumference (r = -0.33),
apolipoprotein B (r = -0.26), systolic blood pressure (r = -0.28), and fasting
serum glucose (r = -0.15); all P<0.0025. [Recall, Reader, that “r” is a
statistical measure ranging from -1 to +1, with the respective extrema (-1 or
+1) indicating complete correlation, either inverse or positive.] In backward
selection linear regression models of telomere length, adiposity measures
were consistently retained in the best models; BMI, waist circumference, hip
circumference, total body fat, and visceral adipose tissue volume were all
inversely associated with telomere length at the nominal P<0.05 level or
lower, independent of age, sex, systolic blood pressure, and fasting serum
lipid, lipoprotein, and glucose concentrations. The negative association of
BMI with telomere length was stronger among younger than older
participants (P for interaction, 0.03).

Conclusions: Individuals with higher total and abdominal adiposity have lower
telomere length, a marker of cellular senescence, suggesting obesity may
 hasten the aging process. Longitudinal studies are required to establish the
causal association of early life adiposity with biological aging.I

The authors of the study were certainly justified in highlighting adiposity as the most
prominent correlate of curtailed telomeres. After all, the reported magnitude of the inverse
correlation between waist circumference and telomere length was considerably greater than
that for fasting glucose concentration and telomere length (r = -0.33 and r = -0.15,
respectively). Nevertheless, both correlations were statistically significant. There is a further
point to consider. Though obesity has attained epidemic proportions in the Americas and
throughout the developed world, individuals of normal bodyweight and body composition
are prevalent in the population. Certainly “normoglycemia” is the norm. That is, most
persons in a given population will not exhibit fasting glucose concentration comparable to
clinically diagnosed diabetics. However, most persons in a given population are not in a
fasting state and nearly none practice perpetual fasting analogous to the Amen Fast. This is
significant insofar as studies such as the one under analysis only entail assessments of
transient fasting in individuals unaccustomed to chronic fasting. Hence, what is revealed
therein is, for example, differing abilities to regulate glucose during relatively brief periods of
dietary restriction and the effect of such restriction on a measure such as telomere length.
This is important and applicable information. However, it would be far more interesting to
assess the effects of systematic, cyclic fasting on such a measure as telomere length. Dr. Lee
and colleagues reported that the mean fasting glucose concentration of the 300+ participants
was ~90 milligrams per deciliter (mg/dL). This is exceedingly high by Amen standards. The
reported range was ~60 to ~290 mg/dL. Admirably, the lower limit is not appreciably
higher than that of the Author as commonly registered at the mid-point of his daily fast (~55
mg/dL). There is an important difference, however. The Author and other Amenites
experience low levels of circulating glucose for 20+ hours daily for years on end.
Presumably, randomly selected study participants do not similarly subject themselves to daily
cyclic fasting of the Amenite sort. Hence, it is justifiable for Adherents to draw stronger
conclusions from such conservative studies insofar as our intervention is far more intensive
and, ex hypothesi, far more efficacious.

Because fasting is the most facile means of suppressing circulating sugar

concentration, because elevated sugar concentration conduces to telomere shortening,
because telomere shortening is associated with accelerated aging, fasting ineluctably emerges
as an efficacious avenue of lifespan prolongation.

Lifespan Length & Telomere Length: Gauging Glycation

The preceding section showed the significance of elevated glucose concentration as a

deleterious determinant of telomere length. Telomere length is an accurate index of aging.
The Amenistic implication of this is clear: Fasting potently suppresses sugar concentration
and consequently facilitates telomere maintenance and lifespan prolongation. Amen
Adherents understand that glycation is a causal factor in the acceleration of aging. We are
also aware that the most efficacious means of mitigating glycation is by curtailing the

ILee M, Martin H, Firpo MA, Demerath EW. Inverse association between adiposity and telomere length: The
Fels Longitudinal Study. American Journal of Human Biology. 2011; 23(1):100-6.
concentration of circulating glucose—that is, by a program of protracted fasting such as that
prescribed by the Amen Protocol. Of interest in this respect is evidence of a correlation
between glycation and telomere length. Specifically, epidemiological evidence indicates that
individuals with impaired glucose tolerance—abnormally elevated glucose concentration
resulting from an impaired ability to metabolize sugar efficiently—exhibit a correlation
between telomere length and the extent of hemoglobin glycation. That is, glucose intolerant
individuals exhibit an inverse correlation between the degree of glycation of their constituent
hemoglobin and the length of telomeres in their leukocytes. This was established by Indian
investigators and published in the journal Atherosclerosis. Though the researchers were
primarily interested in the correlation of cardiovascular complications with telomere length,
they also attended to other important metabolic and pathophysiological outcomes such as
indices of inflammation, oxidation, and, most importantly from our present perspective,
glycemia and glycation. Their undertaking was explained in their Abstract as follows:

Objective: Shortening of telomere length has been reported in several

conditions including Type 2 diabetes and atherosclerosis. The aims of this
study were (1) to assess whether telomere shortening occurs at the stage of
pre-diabetes, i.e., impaired glucose tolerance (IGT) and (2) whether telomere
shortening was greater in Type 2 diabetic subjects with atherosclerotic
plaques.

Methods: Subjects with impaired glucose tolerance (IGT) (n=30), non-diabetic

control subjects (n=30), Type 2 diabetic patients without (n=30) and with
atherosclerotic plaques (n=30) were selected from the Chennai Urban Rural
Epidemiology Study (CURES), an ongoing epidemiological population-based
study. Southern-blot analysis was used to determine mean terminal restriction
fragment (TRF) length, a measure of average telomere size, in leukocyte size,
in leukocyte DNA. Levels of thiobarbituric acid reactive substances
(TBARS), protein carbonyl content (PCO) and high sensitive C-reactive
protein (hs-CRP) were measured by standard methodologies. Carotid intima-
media thickness (IMT) [an indication of arterial thickness] was assessed by
high resolution B-mode ultrasonography.

Results: The mean (+/- SE) [i.e. standard error, an ‘index of imprecision’]
TRF lengths were significantly lower in IGT subjects (6.97 +/- 0.3 kb;
p=0.002) and lower still in Type 2 diabetic subjects without plaques (6.21
+/- 0.2; p=0.0001) and lowest in Type 2 diabetic subjects with
atherosclerotic plaques (5.39 +/- 0.2; p=0.0001) when compared to control
subjects (8.7 +/- 0.5). In IGT subjects, TRF length was positively correlated
to HDL cholesterol and negatively correlated to glycated hemoglobin
(HbA1c), TBARS, PCO, HOMA-IR and IMT. In multiple linear regression
analysis, presence of diabetes, HDL cholesterol and increased TBARS levels
appear as significant determinants of telomere shortening.
 Conclusion: Telomere shortening is seen even at the stage of IGT. Among
subjects with Type 2 diabetes, those with atherosclerotic plaques had greater
shortening of telomere length compared to those without plaques.I

Apart from the item of particular interest to us (i.e. glycation) this study is especially
important insofar as it elegantly establishes that telomere length decreased with increasing
degree of damage done by elevated sugar, from normoglycemic control subjects, to
individuals with impaired glucose tolerance, to persons with diagnosed diabetes, to diabetic
individuals having incurred appreciable atherosclerosis owing ostensibly to elevated
circulating sugar. It was further found that those with impaired glucose tolerance exhibited a
correlation between the degree of dysglycemia and the length of their telomeres—the more
intense the impairment in glucoregulatory ability, the shorter the telomeres. Further, among
this same IGT group, the greater the glycation of their hemoglobin, the shorter their
telomeres.
If glycation is correlated with telomere length, if the same processes that promote
glycation cause telomeres to shorten, it should follow that agents able to inhibit glycation can
conserve telomere length, protecting them from the pathological effects of hyperglycemia.
As shall be discussed in Part I of Volume III, a class of compounds chemically related to
guanidine exerts an appreciable, inhibitory effect against glycation. Such is the case with the
agent aminoguanidine. It was established by Chinese investigators from the Capital Medical
University in Beijing that the application of aminoguanidine to a cellular system suppressed
glycation and promoted the preservation of telomere length. The researchers delineate the
conceptual components of their study as follows:

Background: The accumulation of free radicals and advanced glycation end

products (AGEs) in cells plays a very important role in replicative
senescence. Aminoguanidine (AG) has potential antioxidant effects and
decreases AGE levels. This study aimed to investigate its effect on replicative
senescence in vitro.

Methods: The effects of aminoguanidine on morphology, replicative lifespan,

cell growth and proliferation, AGEs, DNA damage, DNA repair ability and
telomere length were observed in human fetal diploid fibroblasts (2BS).

Results: Aminoguanidine maintained the non-senescent phenotype of 2BS

cells even at late population doubling (PD) and increased cumulative
population doubling by at least 17-21 PDs. Aminoguanidine also improved
the potentials of growth and proliferation of 2BS cells as detected by MTT
assay. The AGE levels of late PD cells grown from early PD in DMEM [a
nutrient medium] containing aminoguanidine decreased significantly
compared with those of late PD control cells and were similar to those of
young control cells. In addition, the cells pretreated with aminoguanidine had
a significant reduction in DNA strand breaks when they were exposed to 200
micromol/L H2O2 [peroxide] for 5 minutes, which indicated that the

IAdaikalakoteswari A, Balasubramanyam M, Ravikumar R, Deepa R, Mohan V. Association of telomere

shortening with impaired glucose tolerance and diabetic macroangiopathy. Atherosclerosis. 2007; 195(1):83-9.
 compound had a strong potential to protect genomic DNA against oxidative
stress. And most of the cells exposed to 100 micromol/L H2O2 had much
shorter comet tails and smaller tail areas after incubation with amino-
guanidine-supplemented [medium], which indicated that the compound
strongly improved the DNA repair abilities of 2BS cells. Moreover, PD55
cells grown from PD28 in 2 mmol/L aminoguanidine retain telomere lengths
of 7.94 kb or 8.12 kb, which was 0.83 kb or 1.11 kb longer than of control
cells.

Conclusion: Aminoguanidine delays replicative senescence of 2BS cells and the

senescence-delaying effect of aminoguanidine appear to be due to its many
biological properties including its potential for proliferation improvement, its
inhibitory effect on AGE formation, antioxidant effect, improvement of
DNA repair ability and the slowdown of telomere shortening.I

Another antiglycative agent is of interest in the present context. Compounds from

the plant belonging to the genus Astragalus have been found to exhibit a four-fold effect:
glucose suppression, glycation inhibition, telomere preservation, and lifespan lengthening.
First, we evaluate evidence of its antiglycative ability. This effect was established by Japanese
investigators from Kumamoto University’s Graduate School of Medical and Pharmaceutical
Sciences. They straightforwardly summarize in their Abstract as follows:

Because advanced glycation end product (AGE) inhibitors such as

pyridoxamine significantly inhibit the development of retinopathy
[pathological dysfunction of the retinal nerve] and neurophathy [pathological
dysfunction of peripheral nerves due to glucose-mediated damage] in the
streptozotocin-induced diabetic rat, treatment with AGE inhibitors is
believed to be a potential strategy for the prevention of lifestyle-related
diseases such as diabetic complications. A crude extract of Astragali Radix
(AR; roots of Astragalus membranaceus) inhibits the formation of N(epsilon)-
(carboxymethyl)lysine (CML) and pentosidine during the incubation of
bovine serum albumin with ribose [a sugar]. In the present study, compounds
were isolated from AR that prevented CML and pentosidine formation.
Astragalosides significantly inhibited the formation of both CML and
pentosidine and astragaloside V had the strongest inhibitory effect among all
of the isolated compounds. These data suggest that AR and astragalosides
may be a potentially useful strategy for the prevention of clinical diabetic
complications by inhibiting AGEs.II

Reiteratively, Astragalus evidently inhibits glycation in an organism in which diabetes has

been experimentally induced. [The ideality of Astragalus’ ability to inhibit pentosidine
accumulation especially shall be evident when we review research presented in Volume III:I

IWang PC, Zhang J, Zhang ZY, Tong TJ. Aminoguanidine delays the replicative senescence of human diploid
fibroblasts. Chinese Medical Journal. 2007; 120(22): 2028-2035.
IIMotomura K, Fujiwara Y, Kiyota N, Tsurushima K, Takeya M, Nohara T, Nagai R, Ikeda T. Astragalosides

isolated from the root of Astragalus radix inhibit the formation of advanced glycation end products. Journal of
Agricultural & Food Chemistry. 2009; 57(17): 7666-72.
indicating an inverse relationship between the amount of the inimical glycating agent
(originating from the effect of superoptimal amounts of the sugar ribose) and maximum
lifespan in mammalian models.]
Independently, it has been shown that Astragalus promotes the prolongation and
preservation of telomeres by augmenting the activity of the enzyme telomerase. This was
established by Iberian investigators from the aptly named “Telomeres and Telomerase
Group” of the Spanish National Cancer Center in Madrid. As their Abstract explains:

...[W]e show that a small-molecule activator of telomerase (TA-65) purified

from the root of Astragalus membranaceus is capable of increasing average
telomere length and decreasing the percentage of critically short telomeres
and of DNA damage in haploinsufficient mouse embryonic fibroblasts
(MEFs) that harbor critically short telomeres and a single copy of the
telomerase RNA Terc gene (G3 Terc(+/-) MEFs). Importantly, TA-65 does
not cause telomere elongation or rescue DNA damage in similarly treated
telomerase-deficient G3 Terc(-/-) littermate MEFs. These results indicate
that TA-65 treatment results in telomerase-dependent elongation of short
telomeres and rescue of associated DNA damage, thus demonstrating that
the TA-65 mechanism of action is through the telomerase pathway. In
addition, we demonstrate that TA-65 is capable of increasing mouse
telomerase reverse transcriptase levels in some mouse tissues and elongating
critically short telomeres when supplemented as part of a standard diet in
mice. Finally, TA-65 dietary supplementation in female mice leads to an
improvement of certain health-span indicators including glucose tolerance,
osteoporosis and skin fitness, without significantly increasing global cancer
incidence.I

Thus, added to the ability of Asgragalus to inhibit glycation is its ability to lengthen telomeres.

We now add to our burgeoning knowledge the ability of Astragalus to lengthen

lifespan. This prolongevity potential of the plant was established in the organism C. elegans
and reported in The Biochemical Journal. The Abstract indicates that:

Late-onset neurodegenerative diseases are characterized by progressive

accumulation of aggregation-prone proteins and global disruption of the
proteostasis network, e.g. abnormal polyQ (polyglutamine) aggregation in
Huntington’s disease. Astragalus membranaceus polysaccharide (astragalan) has
recently been shown to modulate aging and proteotoxic stress pathways.
Using Caenorhabditis elegans models, we now show that astragalan not only
reduces polyQ aggregation, but also alleviates the associated neurotoxicity.
We also reveal that astragalan can extend the adult lifespan of wild-type and
polyQ nematodes, indicating a connection of its anti-aging benefit with the
toxicity-suppressing effect. Further examination demonstrates that astragalan
can extend the lifespan of daf-2 and age-1, but not daf-16, mutant nematodes

IBernardes de Jesus B, Schneeberger K, Vera E, Tejera A, Harley CB, Blasco MA. The telomere activator TA-
65 elongates short telomeres and increases health span of adult/old mice without increasing cancer incidence.
Aging Cell. 2011; 10(4): 604-21.
 of the insulin-like aging and stress pathway, suggesting a lifespan-regulation
signaling independent of DAF (abnormal dauer [a developmental stage]
formation)-2/IGF-1R (insulin-like growth factor 1 receptor), but dependent
on the DAF-16/FOXO (forkhead box O) transcription factor, a pivotal
integrator of divergent signaling pathways related to both lifespan regulation
and stress resistance. We also show that a subset of DAF-16 downstream
genes are regulated by astragalan, including the DAF-16 transcriptional target
gene scl-20, which is itself constitutively up-regulated in transgenic polyQ
nematodes. These findings, together with our previous work on LEA (late
embryogenesis abundant) proteins and trehalsose, provide revealing insight
into the potential of stress and lifespan regulators in the prevention of
proteotoxic disorders.I

Now it is not nearly surprising to the Neophyte why Astragalus extract is included in the
Amen Apothecary and, further, why it is fitting to focus on its effects in our expository
Volume on the Amen Fast & Telomeres: Similar to our sacred Fast the effects of Astragalus
include reduction of glycemia, attenuation of glycation, lengthening (and/or maintenance) of
telomeres and, most momentously, maximization of lifespan potential. Please permit,
pensive Proselyte, the ‘Perennial Pedagogue’ to “pile on”, identifying another agent included
in our Apothecary that is analogously ingested during our Fast and which possesses
telomere-preserving power. This agent is resveratrol. First let us focus on its AGE-inhibiting
ability. This was demonstrated by Drs. Ciddi and Dodda in a paper published in
Pharmacological Reports. As indicated in their Abstract:

Background: Various mechanisms with a complex integrating paradigm have

been implicated in diabetic complications. The present study was aimed to
evaluate the aldose reductase (AR) and advanced glycation end products
(AGEs) inhibitory activity of resveratrol (RSV) and its potential in the
treatment of diabetic complications such as cataract and nephropathy.

Methods: RSV was studied for its inhibitory activity against rat lens AR
(RLAR) and rat kidney AR (RKAR) in vitro along with its ability to inhibit
formation of AGEs. Anticataract activity of RSV was demonstrated using
sugar induced lens opacity model in isolated cattle lens. Furthermore the
involvement of RSV in streptozotocin-induced diabetic nephropathy was
investigated by assessing the key markers of kidney function along with the
formation of AGEs. The potent AR inhibitor, fidarestat was as a standard.

Results: RSV exhibited inhibitory activity against RLAR and RKAR with IC50
values of 4.99 µg/ml (21.9 µM) and 5.49 µg/ml (24.5 µM), respectively. It
also showed a significant inhibition of AGEs formation in vitro. In sugar-
induced lens opacity model, RSV displayed a significant protective effect
preventing opacification and formation of polyols in cattle lens. RSV
significantly improved glycaemic status and renal function in diabetic rats

IZhang H, Pan N, Xiong S, Zou S, Li H, Xiao L, Cao Z, Tunnacliffe A, Huang Z. Inhibition of polyglutamine-
mediated proteotoxicity by Astragalus membranaceus polysaccharide through the DAF-16/FOXO transcription
factor in Caenorhabditis elegans. The Biochemistry Journal. 2012; 441(1): 417-24.
 with a significant decrease in the formation of AGEs in the kidneys.

Conclusions: The results obtained in this study underline the potential of RSV
as a possible therapeutic agent against long-term diabetic complications.I

So resveratrol demonstrably reduces glycation and counters cataract formation and kidney
damage—common complications of diabetes due to the damaging effects of elevated sugar.
Now we turn to telomeres and their response to resveratrol provisionment. We are informed
in the ensuing Abstract in Oncotarget that:

Aging is the predominant risk factor for cardiovascular diseases and

contributes to a considerably more severe outcome in patients with acute
myocardial infarction. Resveratrol, a polyphenol found in red wine, is a
caloric restriction mimetic with potential anti-aging properties which has
emerged as a beneficial nutraceutical for patients with cardiovascular disease.
Although resveratrol is widely consumed as a nutritional supplement, its
mechanism of action remains to be elucidated fully. Here, we report that
resveratrol activates human nicotinamide phosphoribosyltransferase
(NAMPT), SIRT4 and telomerase reverse transcriptase (hTERT) in human
aortic smooth muscle cells. Similar observations were obtained in resveratrol
treated C57BL/6J mouse heart and liver tissues. Resveratrol can also
augment telomerase activity in both human pulmonary microvascular
endothelial cells and A549 cells. Blocking NAMPT and SIRT4 expression
prevents induction of hTERT in human aortic smooth muscle cells while
overexpression of NAMPT elevates the telomerase activity induced by
resveratrol in A549 cells. Together, these results identify a NAMPT-SIRT4-
hTERT axis as a novel mechanism by which resveratrol may affect the anti-
aging process in human aortic smooth muscle cells, mouse hearts and other
cells. These findings enrich our understanding of the positive effects of
resveratrol in human cardiovascular diseases.II

What, the reflective Amenite may inquire, is the relevance of the research related in this
section to fasting? Simple. Glycation and telomere truncation are closely correlated. The
same conditions that catalyze glycation cause curtailment of telomeres. As we have seen,
specific substances such as Astragalus, resveratrol, and aminoguanidine simultaneously
suppress glycation, promote the preservation of telomere length, and promote the
prolongation of life. The most effective inhibitory intervention against glycation is fasting.
Therefore, the Amen Fast is a fascicle means of mitigating telomere truncation and,
accordingly, is an effective anti-aging intervention.

ICiddi V, Dodda D. Therapeutic potential of resveratrol in diabetic complications: In vitro and in vivo studies.
Pharmacological Reports. 2014 Oct; 66(5):799-803.
II
Huang P, Riordan SM, Heruth DP, Grigoryev DN, Zhang LQ, Ye SQ. A critical role of nicotinamide
phosphoribosyltransferase in human telomerase reverse transcriptase induction by resveratrol in aortic smooth
muscle cells. Oncotarget. 2015 Mar 14.
 Lifespan is linked to telomere length. Telomere length is linked to glycation. Inhibiting
glycation ensures telomere protection. Intermittent fasting (enhanced by agents included in
the Amen Apothecary) inhibits glycation and thereby advances the aim of lifespan
prolongation.

Cholesterol Curtails Telomeres; Fasting Suppresses Cholesterol Synthesis: Exploring

Convoluted, Yet Compelling, Correlations

In the Author’s Evolutionary Nutrition, the inimical effects of cholesterol were considered.
Readers interested in these ideas are directed thereto. We shall have further occasion to
discuss cholesterol in Volume III, Part II of AIR—its inclusion is deemed desirable insofar
as The Amen Diet, being thoroughly vegetal, is devoid of exogenous cholesterol. Presently,
we are interested in the interface between cholesterol, telomeres, aging, and fasting. First, it
is important to relate to the Reader the fact that chemically countering cholesterol synthesis
has been found to promote the preservation of telomeres. The molecular modulation of
cholesterol synthesis was effectuated by exposing animals to a widely prescribed
pharmaceutical drug belonging to the statin family. Mechanistically, statins exert their effect
by inhibiting enzymes involved in the synthesis and uptake of cholesterol. The most marked
molecular effect, and the most meaningful from a clinical perspective is the suppression of
serum cholesterol. We shall cite two relevant studies—both observational, epidemiological
investigations. The first was undertaken by French scientists from the Université de
Bourgogne in Dijon and reported in the journal Atherosclerosis. The Abstract appears as
follows:

Background:…We investigated the relationship between prior statin therapy

and leukocyte telomere length (LTL), as well as their interaction with
potential new biomarkers of oxidative deoxyribonucleic acid (DNA) lesions
and reactive oxygen species-induced inflammation.

Methods & Results: From patients admitted for an acute myocardial infarction,
LTL was assessed by quantitative polymerase chain reaction (Q-PCR), and
leukocyte Finkel-Biskis-Jinkins osteosarcoma (FOS) and 8-oxoguanine DNA
glycosylase (OGG1) messenger ribonucleic acid (mRNA) levels were
measured by retrotranscription Q-PCR. Patients under prior chronic statin
therapy were older, their mean LTL was longer than patients not under statin
therapy (1.29 +/- 0.11 vs. 1.25 +/- 0.11 T/S ratio, p=0.008). In contrast,
FOS and OGG1 mRNA levels were similar for the 2 groups. LTL decreased
with increasing age, FOS, and OGG1 mRNA levels. Statin therapy remained
associated with longer LTL, even after adjustment for confounding factors
(p=0.001), and in younger patients (! 64 y). Even in groups matched for
propensity scores for statin use, LTL was markedly longer in patients under
statin therapy.

Conclusions: Our observational study showed that statin therapy was

associated with longer LTL. These data bring to light opportunities to
identify new targets for early primary preventive treatment strategies.
 Moreover, our study raised FOS and OGG1 as new relevant biomarkers of
LTL.I

Again we reiterate the consistent finding that telomere length lessened with age. Next we
consider a more incisive epidemiological study undertaken by Italian investigators from
Naples University’s Department of Internal Medicine. Their investigation is more incisive
and informative insofar as it assessed the effect of statins on the activity of the enzyme
telomerase in addition to determining the effect of the agent on telomere length. Their
abstract states that:

Recent evidence suggests a link between statins and telomere biology.

Whether statin treatment may modulate telomerase activity and affect
telomere erosion rate is unknown. We aimed at investigating the potential
impact of statin therapy on peripheral blood mononuclear cells telomerase
activity, its implication on LTL variability, and its association with telomere
shortening rates with aging. The cross-sectional study was conducted in 230
subjects (age range: 30-86 y) stratified according to statins treatment. LTL
was measured by quantitative polymerase chain reaction and telomerase
activity by a PCR-ELISA protocol. Subjects on statin treatment showed
higher telomerase activity independently of multiple covariates, including age,
gender, smoking habits, lipid, systemic inflammation, glucose, and blood
pressure levels (P=0.019). Indeed, subjects on statin treatment showed
significantly lower telomere erosion along with aging. Every 1 yr increment in
age, LTL decreases by 0.058 kb [kilobases, i.e. 103 nucleotide bases] in no
statin and 0.033 kb in statin groups, respectively, as well as the major
difference in telomere attrition between groups was found after the age of 65
yr (P<0.0001). In summary, statins, modulating telomerase activity, affect
telomere erosion along with aging.II

Not surprisingly, the Italian investigators confirmed the finding that telomere length lessens
with age. Importantly, they also found that statin treatment reduced the rapidity of telomere
truncation with age, an effect attributable to the activation of telomerase.
A more compelling case for the anti-aging efficacy of statins could be made if it were
established that the agent increases lifespan in an animal species. This has indeed been
established in the organism Drosophila in a study executed by Dr. Stephen Spindler and
colleagues. Their abstract indicates that:

Statins such as simvastatin are 3-hydroxy-3-methylglutaryl coenzyme A

(HMG-CoA) reductase inhibitors and standard therapy for the prevention
and treatment of cardiovascular diseases in mammals. Here we show that
simvastatin significantly increased mean and maximum lifespan of Drosophila
melanogaster...and enhanced cardiac function in aging flies by significantly

ISaliques S, Teyssier JR, Vergely C, Lorgis L, Lorin J, Farnier M, Donzel A, Sicard P, Berchoud J, Lagrost AC,
Touzery C, Ragot S, Cottin Y, Rochette L, Zeller M. Circulating leukocyte telomere length and oxidative stress:
a new target for statin therapy. Atherosclerosis. 2011; 219(2): 753-60.
IIBoccardi V, Barbieri M, Rizzo MR, Marfella R, Esposito A, Marano L, Paolisso G. A new pleiotropic effect of

statins in elderly: modulation of telomerase activity. FASEB Journal. 2013, June 7.

 reducing heart arrhythmias and increasing the contraction proportion of the
contraction/relaxation cycle. These results appeared independent of internal
changes in ubiquinone or juvenile hormone levels. Rather, they appeared to
involve decreased protein prenylation. Simvastatin decreased the membrane
association (prenylation) of specific small Ras GTPases [signaling molecules]
in mice. Both farnesyl...and type 1 geranylgeranyl transferase...inhibitors
increased Drosophila lifespan. These data are the most direct evidence to
date that decreased protein prenylation can increase cardiac health and
lifespan in any metazoan species, and may explain the pleiotropic (non-
cholesterol related) health effects of statins.I

Cholesterol is a complex chemical whose consumption, synthesis, signaling, and excretion

exert innumerable effects on the physiology of the body. What the research we have
reviewed thusfar reveals is that agents which ordinarily inhibit the synthesis of cholesterol
(specifically statins) are able to extend the lifespan of multicellular organisms via avenues
involving alterations of the manner in which membrane lipids are molecularly modified (e.g.
by farneyslation and geranylation). There are undoubtedly other unobserved physiological
alterations induced by the inhibition of cholesterol synthesis that explain the ability of this
intervention to extend lifespan.

In order to augment our confidence that cholesterol is an active accomplice in the

aging process, it is advantages to ascertain whether other established anti-aging interventions
alter cholesterol metabolism in an inhibitory manner. Importantly, investigations
encompassing two prominent pillars of our Tetractys—cyclic fasting (CF) and caloric
restriction (CR) aid us in this agenda. First, we shall apprise ourselves of an investigation by
Italian researchers from the Università di Cagliari. Reported in the journal Mechanisms of Aging
& Development, the abstract explains that:

The effects of ageing on the metabolism of cholesterol were examined in

three different organs (liver, aorta, and brain) of 6-, 12- and 24-month old
male Sprague-Dawley rats. Ageing was associated with a significant increase
in intracellular cholesterol esters in all three organs. Steady state mRNA
levels of multidrug resistance protein (MDR) and acylCoA:cholesterol acyl
transferase (ACAT), enzymes involved in cholesterol import and
esterification, were also increased. By contrast, expression of mRNA for
neutral cholesterol ester hydrolase (nCEH) and caveolin-1, proteins involved
in cholesterol ester hydrolysis and export, were significantly reduced. Dietary
restriction is the only intervention shown to extend lifespan and retard age-
related declines in function in mammals. To further explore the possible
correlation between changes in cholesterol esterification and aging, we
analyzed cholesterol metabolism in liver, aorta, and brain of aged rats
exposed to two dietary restriction regiments: intermittent (alternate-day)
fasting (IF) and food intake restriction (60% of ad libitum feeding). Both

ISpindler SR, Li R, Dhahbi JM, Yamakawa A, Mote P, Bodmer R, Ocorr K, Williams RT, Wang Y, Ablao KP.
Statin treatment increases lifespan and improves cardiac health in Drosophila by decreasing specific protein
prenylation. PLoS One. 2012; 7(6):e39581.
 dietary regimens attenuated the age-related changes in cholesterol esters and
in the expression of genes involved in cholesterol metabolism. These results
provide evidence that distinctive age-associated changes in intracellular
cholesterol metabolism occur in rats. Furthermore, these modifications can
be partially reversed by dietary restriction, a condition known to affect the
ageing process. Age-related changes in cholesterol metabolism may play a
role in triggering and/or aggravating senescence-related disorders
characterized by altered cholesterol metabolism.I

Overlooking the overzealous investigators’ erroneous remark that “Dietary restriction is the
only intervention shown to extend lifespan...”, let us rapidly recapitulate the researchers’
most important results. They found that: (I) cholesterol concentration increases with age in
several organs (II) enzymes that effectuate cholesterol uptake increase with age (III) enzymes
that effectuate cholesterol expurgation decrease with age and, finally, (IV) both cyclic fasting
and caloric restriction retard the age-related increase in cholesterol accumulation and avert
the age-associated alteration of enzymes involved in cholesterol metabolism.
While the above study employed the “alternate-day” style of cyclic fasting, a joint
team of Belgian and Algerian investigators undertook an analysis of the effect of fasting for
15 hours daily on cholesterol concentration in a desert-adapted rodent commonly called the
sand rat (Psammomys obesus). Interestingly, this experimental organism was utilized by
investigators owing to its propensity to incur obesity under conditions of caloric excess. This
feature makes it a fitting subject for facile physiological comparisons to the human animal.
As the abstract in the International Journal of Molecular Medicine indicates:

The present report concerns several post-mortem variables examined in sand

rats that were either maintained on a vegetal diet (control animals) or
exposed first during a 20-day transition period to a mixed diet consisting of a
fixed amount of a hypercaloric food and decreasing amounts of the vegetal
food and then to a 30-day experimental period of exposure to the
hypercaloric food. During the latter period, all animals were either given free
access to food or fasting daily for 15 h, i.e. from 5:00 p.m. to 8:00 a.m. The
body weight, liver wet weight, pancreas wet weight, plasma glucose and
haemoglobin A1c concentration, plasma insulin concentration, insulinogenic
index, insulin resistance HOMA, plasma cholesterol and triglyceride
concentration, liver triglyceride and phospholipid content were measured.
Pancreatic islet (insulin GLUT2) and liver (lipid droplets) histology were also
examined. The main findings consisted in a lower body weight of fasting
than non-fasting animals, a higher liver weight in non-diabetic and diabetic
rats than in control non-fasting (but not so in fasting) animals, a decrease of
pancreas weight in non-diabetic and diabetic as distinct from control animals,
a fasting-induced decrease in plasma glucose, plasma insulin and insulin
resistance HOMA [an index of the “metabolic syndrome”], plasma
cholesterol and triglyceride concentration and triglyceride liver content.II

IMulas MF, Demuro G, Putzolu M, Cavallini G, Donati A, Bergamini E, Dessi S. Dietary restriction
counteracts age-related changes in cholesterol metabolism in the rat. Mechanisms of Ageing & Development. 2005;
126(6-7): 648-54.
IIBelkacemi L, Selselet-Attou G, Bulur N, Louchami K, Sener A, Malaisse WJ. Intermittent fasting modulation
 Excursus: Though a 15-hour fast is not exceedingly abstemious by Amenite
standards, these scientists were seeking to replicate the rather relaxed rigor of the
Ramadan fast. As Ramadan proscribes partaking of food or drink during daylight
hours, this may merely mean a fast of 8-10 hours or less, as it is common for the
adherents of Islam to eat immediately upon sunset and subsequently, preceding
sunrise.

Thus we see that cyclic fasting for a period far less lengthy than our Amenite interval of 23
hours is sufficient to suppress circulating cholesterol concomitant with a reduction in
circulating glucose concentration. An investigation undertaken a quarter century ago
established the ability of fasting to suppress cholesterol more convincingly. It is of more
interest to Amenites for two reasons: (I) the duration of the fast was a full 24-hr day—clearly
comparable to the ideal Amen interval of 23+ hours and (II) the investigators assessed the
activity of the enzyme HMG-CoA reductase in addition to absolute cholesterol
concentration. The tandem of Scottish scientists reported their principal findings in the
Biochemistry Journal thusly:

Long-term starvation (24h) resulted in a marked (80%) decrease in total

enzyme activity as well as in the [effective activity of the enzyme] (from 79.3
± 2.4% in control fed animals to 25.2 ± 3.0% in starved animals). This
observation confirms the inference from our previous studies on diabetic rats
(Easom & Zammit, 1985), namely that the phosphorylation state of HMG-
CoA reductase can change chronically in response to altered physiological
conditions. As a result of the combined effects of the changes in total activity
of HMG-CoA reductase and of its phosphorylation state, the expressed
activity was diminished to about 6% of control values after 24 h starvation.I

The Reader is reminded that the direction of this discourse is dictated determinately by the
observation that pharmacologic inhibition of the enzyme HMG-CoA reductase with statins
promoted the preservation of telomere length. Our discursive detour into a discussion of
diet was deemed fitting insofar as fasting has been found to concomitantly curtail cholesterol
concentration and promote telomere preservation, probably owing to its suppression of
circulating glucose. Exactly why cholesterol concentration is correlated with telomere length
is obscure. It is known, however, that insulin exerts an influence on the endogenous
production and processing of cholesterol. Insulin is, of course, elaborated especially in
response to elevated glucose. Thus, perhaps cholesterol is merely a passive player in the
pernicious process leading to telomere truncation whereas high circulating sugar is the
sinister central player in accelerating the shortening of telomeres and, therewith, aging. It
seems that we cannot be quite sure entirely on the basis of available evidence. However, the
cluster of correlations between indices of aging, telomere length, glucose concentration,
fasting, cholesterol, and the conditions controlling cholesterol concentration in the body is

of the diabetic syndrome in sand rats. III. Post-mortem investigations. International Journal of Molecular Medicine.
2001; 27(1): 95-102.
I
Easom RA, Zammit VA. Acute effects of starvation and treatment of rats with anti-insulin serum, glucagon
and catecholamines on the state of phosphorylation of hepatic 3-hydroxy-3-methylglutaryl-CoA reductase in
vivo. Biochemistry Journal. 1987; 241(1):183-8.
too compelling to ignore, despite its complexity and lack of clarity. In this context, our
appreciation of an illuminating epidemiological investigation by British and Saudi scientists is
well warranted. This study established an interrelationship between telomere length,
glycemia, and cholesterol concentration. Specifically, diabetics (clearly characterized by
elevated glucose concentration) exhibited elevated cholesterol and shorter telomeres than
comparable controls devoid of diabetes. As their Abstract explains:

South Asians have a higher risk of type 2 diabetes mellitus (T2DM) and
cardiovascular disease (CVD) than white Caucasians, for a given BMI.
Premature biological aging, assessed by reduction in telomere length (TL),
may be mediated by factors resulting from altered metabolic profiles
associated with obesity. We hypothesise that ethnicity and metabolic status
represent detrimental factors contributing to premature biological aging.
Therefore we assessed TL in two South Asian, age and BMI- matched
cohorts (T2DM (n = 142) versus non-T2D (n = 76)) to determine the effects
of BMI, gender and CVD profile on biological aging. Genomic DNA was
obtained from the UKADS cohort [a sample of subjects selected for
epidemiological analysis]; biochemical and anthropometric data was collected
and TL was measured by quantitative real-time PCR. Our findings indicated
a gender-specific effect with reduced TL in T2Dm men compared with non-
T2DM men (P = 0.006). Additionally, in T2DM men, TL was inversely
correlated with triglycerides and total cholesterol (r = -0.419, P < 0.01; r = -
0.443, P < 0.01). In summary, TL was reduced amongst South Asian T2DM
men and correlated with triglycerides and total cholesterol. This study
highlights enhanced biological ageing among South Asian, T2DM men,
which appears to be tracked by changes in lipids and BMI, suggesting that
raised lipids and BMI may directly contribute to premature aging.I

To reiterate, whether raised cholesterol contributes directly or indirectly to attenuated

telomere length is unascertainable presently. Its involvement is undeniable, however, if the
accumulated evidence has been interpreted accurately by the Author and other primary
investigators. Our confidence in the correctitude of our interpretation of the evidence is
increased by a conceptually complementary study. In this epidemiological investigation
undertaken by Italian scientists, subjects were selected on the basis of cardiovascular
condition and chronological age. To our approval and appreciation, among their uppermost
outcomes of interest was telomere length and fasting glucose concentration. Both measures
were closely correlated among myocardial infarct sufferers—that is, among subjects
succumbing to the most serious clinical effect of elevated cholesterol. As they explain in
their Abstract.

We performed a cross-sectional study to examine the difference in leukocyte

telomere length among three groups of subjects: patients with type 2 diabetes

IHarte AL, da Silva NF, Miller MA, Cappuccio FP, Kelly A, O’Hare JP, Barnett AH, Al-Daghri NM, Al-Attas
O, Alokail M, Sabico S, Tripathi G, Bellary S, Kumar S, McTernan PG. Telomere length attrition, a marker of
biological senescence, is inversely correlated with triglycerides and cholesterol in South Asian males with type 2
diabetes mellitus. Experimental Diabetes Research. 2012: 895185.
 mellitus without history of previous myocardial infarction (Type2DM),
patients with type 2 diabetes mellitus with evidence of previous myocardial
infarction (Type2DM+MI), and healthy control subjects (CTR). The main
objective of the present study is to investigate differences in telomere length
between the studied groups of subjects, with the aim to clarify if telomere
length could be a reliable marker associated with MI in Type2DM patients.
Secondary end point is the identification of associations between leukocyte
telomere length and selected variables related to glycemic control, pro-
inflammatory status and lipidic profile.

Research Design & Methods: A total of 272 elderly subjects, 103 Type2DM
(mean age 70 ± 4 years, 59% males), 65 Type2DM+MI elderly (mean age 68
± 7 years, 68% males), and 104 CTR (mean age 69 ± 7 years, 50% males)
were studied. Telomere length, defined as T/S (Telomere-Single copy gene
ratio), was determined in leukocytes by quantitative real-time polymerase
chain reaction (real-time PCR)-based assay. Moreover, we assessed: (1) high
sensitive C reactive protein (hsCRP), fibrinogen and plasminogen-activator
inhibitor-1 (PAI-1) as inflammatory markers; (2) fasting glucose, insulin,
glycated haemoglobin (HbA1C) and waist-to-hip ratio as markers of glycemic
control; (3) total-cholesterol, and triglycerides as markers of lipidic profile, in
all sample population. The use of statins and sulfonylurea, as well as the
presence of some relevant diabetes complications (nephropathy and
retinopathy) were also assessed.

Conclusion: Type2DM+MI elderly patients have leukocyte telomere lengths

shorter than those of Type2DM (without MI) and healthy [controls].
Moreover, glucose HbA1C and waist-to-hip ratio, variables related to
glycemic control, showed a significant inverse correlation with leukocyte
telomere length.I

This conclusion is corroborated by a comparable study concerning the correlative

consequences of elevated glucose concentration and cardiovascular pathology on telomere
length. This investigation is especially illuminating insofar as it shows how the severity of
telomere shortening increases stepwise with the increase in clinical severity of the conditions
under consideration—that is, diabetic persons with cholesterol-containing atherosclerotic
plaques had shorter telomeres than diabetics devoid of such plaques, who had shorter
telomeres than individuals with impaired glucose tolerance, who, in turn, had shorter
telomeres than controls. As their abstract in Atherosclerosis explains:

Shortening of telomere length has been reported in several conditions

including Type 2 diabetes and atherosclerosis. The aims of this study were (1)
to assess whether telomere shortening occurs at the stage of pre-diabetes, i.e.,

IOivieriF, Lorenzi M, Antonicelli R, Testa R, Sirolla C, Cardelli M, Mariotti S, Marchegiani F, Marra M,

Spazzafumo L, Bonfigli AR, Procopio A. Leukocyte telomere shortening in elderly Type2DM patients with
previous myocardial infarction. Atherosclerosis. 2009; 206(2):588-93.
 impaired glucose tolerance (IGT) and (2) whether telomere shortening was
greater in Type 2 diabetic subjects with atherosclerotic plaques.

Methods: Subjects with impaired glucose tolerance (IGT) (n=30), non-diabetic

control subjects (n=30), Type 2 diabetic patients without (n=30) and with
atherosclerotic plaques (n=30) were selected from the Chennai Urban Rural
Epidemiology Study (CURES), an ongoing epidemiological population-based
study. Southern-blot analysis was used to determine mean terminal restriction
fragment (TRF) length, a measure of average telomere size, in leukocyte
DNA. Levels of thiobarbituric acid reactive substances (TBARS), protein
carbonyl content (PCO) and high sensitive C-reactive protein (hs-CRP) were
measured by standard methodologies. Carotid intima-media thickness (IMT)
was assessed by high resolution B-mode ultrasonography.

Results: The mean (+/-S.E.) TRF lengths were significantly lower in IGT
subjects (6.97+/-0.3 kb; p=0.002) and lower still in Type 2 diabetic subjects
without plaques (6.21 +/-0.2; p=0.00001) and lowest in Type 2 diabetic
subjects without atherosclerotic plaques (5.93 +/-0.2; p=0.0001) when
compared to control subjects (8.7+/-0.5). In IGT subjects, TRF length was
positively correlated to HDL cholesterol and negatively correlated to glycated
hemoglobin (HbA1c), TBARS, PCO, HOMA-IR and IMT. In multiple linear
regression analysis, presence of diabetes, HDL cholesterol and increased
TBARS levels appear as significant determinants of telomere shortening.

Conclusion: Telomere shortening is seen even at the stage of IGT. Among

subjects with Type 2 diabetes, those with atherosclerotic plaques had greater
shortening of telomere length compared to those without plaques.I

Fasting glucose concentration, impaired post-absorptive glucose concentration, age,

telomere length, clinical consequences of chronic hypercholesterolemia—the above
epidemiological analyses again link them all. Additional elements are hereby added to our
explanatory framework, clarifying a complex issue considerably.
In aggregate, the implications of the investigations encountered in this section are as
follows. First, heightened levels of cholesterol compromise longevity. This is apparent from
two independent evidentiary angles—the extension of metazoan lifespan by an agent that
inhibits cholesterol concentration in clinical applications and the protection of telomeres
from shortening by statins—the same cholesterol-suppressing substance that attenuates
aging in the organism C. elegans. Insofar as we are aware of the molecular mechanism by
which statins exert their principal effects (i.e. inhibition of the enzyme HMG-CoA reductase)
we are confident in contending that fasting can conduce to lifespan prolongation through
this same mechanism—that is, prevention of telomere truncation via cholesterol curtailment.
This reasoning rests on the findings that fasting attenuates expression of the cholesterol-
enhancing enzyme, HMG-CoA reductase. Further, on the strength of the discovery by Drs.
Easom and Zammit determining that fasting for the duration of a day diminishes the

IAdaikalkoteswari A, Balasubramanyam M, Ravikumar R, Deepa R, Mohan V. Association of telomere

shortening with impaired glucose tolerance and diabetic macroangiopathy. Atherosclerosis. 2007; 195(1): 83-9.
enzymatic activity of HMG-CoA by upwards of eighty percent, we are confident that the
distinctive nature of the Amen Fast is particularly productive of cholesterol curtailment and
consequent telomere preservation and, ultimately, lifespan lengthening.

The Amen Fast conceivably conduces to lifespan prolongation by inhibiting the

enzymatic synthesis of cholesterol, which (somehow) spares telomeres from shortening.

Amenistic Meditation May Mitigate Mortality by Maintaining Telomere Length

The Author deems it appropriate to mention meditation in this section of our Treatise for
two principal reasons (I) Amen Meditation is executed only during the Fasting Phase and (II)
the particular posture of meditation prescribed by our Protocol promotes telomere
preservation and, perforce, lifespan prolongation.I To be sure, the twice-daily Amen Exercise
Sessions are executed solely during the fasted state as well and, as we shall see in Volume IV,
exercise ordinarily elicits elongation of telomeres. However, as Amen Meditation is executed
intermittently throughout the Amen Fast and because the psycho-spiritual vacuity that our
modality of meditation is meant to induce mirrors the visceral vacuity that fasting facilitates,
the convergent effect of fasting and meditation on telomere elongation is altogether topical
to this Treatise. It must be mentioned that Amen Meditation entails two fundamental forms,
essentially erect or recumbent, standing or lying. The lying form we designate with either the
Yoga-inspired Sanskrit signifier Sava-Sana (i.e. “Corpse Posture”) or, preferentially, with the
Ancient Egyptian appellation:

Irw sdjr [Phonetically Arew Sedjer] (“Lying Form”)

The standing form we designate with either the Jainist-inspired Sanksrit signifier Kayotsarga
(i.e. “Forgetting the Body”) or, preferentially, with the Ancient Egyptian appellation:

Irw aha [Phonetically Arew Aha] (“Standing Form”)

Excursus: As we shall discuss in further detail in Volume VI (On the Nature of Nous:
Mind & the Mitigation of Mortality), both forms of Amen Meditation have specific,
practical benefits. When one is able to invest appreciable time and energy to the aim
of enhancing self-awareness through asceticism, the extended intervals of ‘Auto-
Ataraxy’II (a Nunistic neologism) or meditation that Arew Sedjer permit are especially

ISjögren P, Fisher R, Kallings L, Svenson U, Roos G, Hellenius ML. Stand up for health—avoiding sedentary
behaviour might lengthen your telomeres: secondary outcomes from a physical activity RCT in older people.
British Journal of Sports Medicine. 2014; 48(19): 1407-9.
II‘Auto-Ataraxy’ is a Nunistic neologism. Whereas “auto” is a Greek term signifying “self”, “ataraxy” derives

from !!"#"$%" (ataraksia) impassiveness, < ! + !"#&''()* (tarassein) to disturb, stir up. In invoking this term
the Author intends to convey the connotation that Amen Meditation aims at a state of self-imposed stillness,
sedateness, and emotional equanimity.
 ideal. It is also ideal for Anchorite Initiates as it facilitates the prolonged practice
essential to attaining the mental mastery, which is the ultimate aim of Amen
Meditation. Conversely and complimentarily, Arew Aha is ideal for easily integrating
intermittent meditation sessions in short intervals throughout the length of long
days in almost any environment affording sufficient solitude and quietude. Though
life in the modern world may make such solitude and quietude increasingly
infrequent, the nature of standing posture of Arew Aha necessitates minimal space
and no accoutrements for sitting or lying—it can be conducted clandestinely in the
quiet corner of any cubicle or small cell or equally well in an expansive wooded area,
the shore of a lake, or an isolated island retreat in the middle of nowhere. Indeed, an
aim of Amen Meditation is to make Nowhere accessible from Anywhere.

In a surprising study conducted by Swedish scientists and published in the British Journal of
Sports Medicine, it was found that whereas exercise was not efficacious in extending the
telomeres of aged individuals, reducing the amount of time spent sitting was in fact effective.
This seems to suggest that simply standing stimulates telomere elongation. The Summary of
the study is as follows:

Background: Telomere length has been associated with a healthy lifestyle and
longevity. However, the effect of increased physical activity on telomere
length is still unknown. Therefore, the aim was to study the relationship
between changes in physical activity level and sedentary behaviour and
changes in telomere length.

Methods: Telomere length was measured in blood cells 6 months apart in 49,
68-year-old, sedentary, overweight individuals taking part in a randomised
controlled physical activity intervention trial. The intervention group received
individualised physical activity on prescription. Physical activity was
measured with a 7-day diary, questionnaires and a pedometer. Sitting time
was measured with the short version of The International Physical Activity
Questionnaire.

Results: Time spent exercising as well as steps per day increased significantly
in the intervention group. Reported sitting time decreased in both groups.
No significant associations between changes in steps per day and changes in
telomere length were noted. In the intervention group, there was a negative
correlation between changes in time spent exercising and changes in telomere
length (rho=-0.39, p=0.07) [Though not statistically significant as indicated
by the probability value (p) greater than 0.05.] On the other hand, in the
intervention group, telomere lengthening was significantly associated with
reduced sitting time (rho=-0.68, p=0.02).

Conclusions: Reduced sitting time was associated with telomere lengthening in

blood cells in sedentary, overweight 68-year-old individuals participating in a
6-month physical activity intervention trial.

Importantly, other investigations have established the ability of exercise (especially in

younger persons) to lengthen or preserve telomeres. In a chain of reasoning that the Author
openly appreciates, a team of Russian researchers interested in novel drug development
speculated that exercise induces catabolism of the carnosine stored in muscle.I Carnosine is
comprised of the amino acids alanine and histidine. Histidine, the Reader is reminded,
contains the anti-glycative imidazolium functional group. We have already seen that
suppression of glycation correlates with telomere preservation. Thus, this is one logical
linkage between exercise and extension of telomeres. There is a more proximate linkage. An
elegant in vitro study by Beijing-based scientist Dr. Shao established that exposing cells to
carnosine itself was able to forestall shortening of telomeres in human fibroblasts.I
Extending these insightful analyses in an Amenistic manner, it is possible that the prolonged
fasting that our Protocol prescribes promotes catabolism of carnosine stored in skeletal
muscle, and that this effect is enhanced not only by the inclusion of the amino acids alanine
and histidine in the Amen Amino Aliksir, it is also augmented by the additive effects of
exercise and the Irew Aha mode of meditation. This complex cluster of conditions
conceivably conduces to lifespan extension via consequent telomere extension—reasoning
refreshingly redolent of Selective Summation.

A fundamental form of Amen Meditation, Arew Aha, entails standing. Amen

Meditation is executed exclusively during the Amen Fast. Studies support the supposition
that standing promotes the preservation of telomeres. Amen Meditation may mitigate
mortality by means of maintaining telomere length.

IBabizhayev MA, Vishnyakova KS, Yegorov YE. Hormone-brain-aging relationships, broadly reactive with
imidazole-containing dipeptides: targeting of telomere attrition as an aging biomarker and dynamic telomerase
activity flirting. Journal of Basic Clinical Physiology & Pharmacology. 2014. Aug 13.
IShao L, Li QH, Tan Z. L-carnosine reduces telomere damage and shortening rate in cultured normal

fibroblasts. Biochemical & Biophysical Research Communications. 2004; 324(2):931-6.