The document discusses healthy aging and the effects of nutraceuticals on gene expression. It covers several topics related to aging including the hallmarks of aging like inflammation, glycation, methylation, and oxidative stress. It provides information on maintaining healthy aging through diet, nutraceutical supplementation, and exercise. Tips are given on anti-inflammatory herbs, foods rich in omega-3, and ways to reduce advanced glycation end products and chronic inflammation.

1. Healthy aging and effects of
nutraceuticals on gene
expression
Maria Vranceanu
University of Medicine and Pharmacy Cluj Napoca
Romania

2.  Understanding the hallmarks of aging
 Articulating the many questions about the future of aging for individuals and for
societies
 Understanding the mechanism behind aging procress
 What happens to our body built with age?
 Evaluating current trends, potential changes and challenge in speculating about
aging’s implications for individual experiences.
 Understanding the promise held by antiaging medicine and study of aging within
and across disciplines
 Tips about healthy eating, nutraceuticals and exercising to develop a healtheir and
better habits of aging
 Imaging onself as a future aging person and as a part of an aging study cohort!
Learning Objectives

3. Outline
 Introduction: aging and healthy aging
 The Hallmarks of aging
 Inflammation
 Glycation
 Methylation
 Oxidtaive stress
 Conclusion

4. Ageing or aging =the process of becoming older.
represents the accumulation of changes in a human being over
time,encompassing physical, psychological, and social changes.
Aging is among the greatest known risk factors for most human
diseases: of the roughly 150,000 people who die each day
across the globe, about two thirds die from age-related causes.
Physiological or
normal
does not require therapeutic
interventions
Pathological
Requires therapeutic
interventions
Hayflick (1998) argues that the aging process (primary aging)
itself can be differentiated from processes that cause disease
(secondary aging)

5. Healthy aging
the development and maintenance of optimal mental, social, and physical well-
being and function in older adults. This will most likely be achieved when
communities are safe, promote health and well-being and use health services and
community programs to prevent or minimize diseases

6. The hallmarks of aging
Lopez-Otic C et al. Cell.2013 Jun 6;153(6): 1194-1217

8. Acute inflammation
begins within seconds to minutes following the injury of tissues.
The damage may be purely physical, or it may involve the activation of an immune response.
Three main processes occur:
• Increased blood flow due to dilation of blood vessels (arterioles) supplying the region
• Increased permeability of the capillaries, allowing fluid and blood proteins to move into the
interstitial spaces
• Migration of neutrophils (and perhaps a few macrophages) out of the capillaries and venules
and into interstitial spaces

9. Chronic Inflammation
~
Overload of proinflammatory
cytokines in the body which
leads to catastrophic effects
when released sistematically
in the body

10. Chronic Inflammation
SKIN AGING
KIDNEY AND LIVER DAMAGE
ARTHROSIS
OSTEOPOROSIS
SARCOPENIA
BRAIN AGING
OBESITY
INSULIN RESISTANCE
TUMORS
ATHEROSCLEROSIS-MI
IMMUNOSENESCENCE

11. The key to successful aging is firstly linked to the need to decrease chronic inflammation,
without compromising an acute response in case of exposure of the organism to pathogens.

12. Clinical monitoring
Get tested for chronic inflammation
• Omega 3 screening
-Ratio omega 6/omega3; AA/EPA and AA/DHA
reference parameters: OMS-4:1 antiaging medicine 3:1 and
2:1OMS, 1:1 AM
• C reactive protein
Reference parameters: OMS 0-10 mg/L
Antiaging medicine: < 1 mg/L
• hsCRP-stronger predictor of heart diseases than LDL cholesterol
References parameters in antiaging medicine < 0.8mg/L
• Fibrinogen
Reference parameters: OMS 150-400mg/dL
antiaging medicine < 280mg/dL

13. Proinflammatory
cytokines
Optimal anti-inflammatory
range
TNF-α
IL-1B
IL-6
IL-8
< 8.1 pg/mL
<15 pg/mL
<12 pg/mL
<32 pg/mL

14. Controlling excess inflammation
Fat classification
Unsaturated fats
SATURATED
TRANS-trans unsatureted
INTERESTERIFIED
Monounsaturated
OMEGA 7
Palmitoleic acid
Vaccinic acid
OMEGA 9
Oleic acid
Erucic acids
Polyunsaturated
Omega 3-ALA, EPA, DHA
Omega 6-Linoleic acid LA, gamma linolenic acid
GLA, Arachidonic acid AA

15. Omega
3
Inflammation
Plt aggregation
Omega
6
Inflammation
Plt aggregation

16. Omega 6/ Omega 3
Too few omega 6 EFA’s could lead to impaired cell growth and
differentiation, increased risk of bleeding and impaired
immune function.
Too few omega 3’s, on the other hand, could lead to increased
risk of cancer, blood clots and inflammatory and autoimmune
diseases.
It is important to have a balance between the different kinds
of eicosanoids

17. DHA / EPA
Which is the role of DHA?
Which is the role of EPA?
DHA / EPA
Which is the role of DHA?
Which is the role of EPA?

18. CHRONIC
DISEASES RISK FACTORS COMMENTS OMEGA-3 OMEGA-6
Cardiovascular
Diseases
Arrhythmias
(irregular heart
beat)
Causes sudden
cardiac death
Lowers Increases
Thrombosis (clot)
Leads to
myocardial
infarction or
stroke
Lowers Increases
Atherosclerotic
plaque
Leads to
atherosclerosis
Lowers Increases
HDL Good cholesterol Increases Lowers
LDL Bad cholesterol Lowers Increases
Triglycerides
Cardiovascular
risk factor
Lowers Increases
Inflammatory
Responses
IL-1 (Interleukin 1)
Inflammation
response
Lowers Increases
IL-6 (Interleukin 6)
Inflammation
response
Lowers Increases
CRP (C-reactive
protein)
Inflammation
response
Lowers Increas

19. Food rich in omega 3
• Mackerel 5134 mg/100 grams
• Salmon 2260 mg/100 grams
• Cod liver oil 2664 mg in a single tablespoon
• Herring 1729 mg/100 grams
• Sardines 1480 mg/100 grams
• Anchovies 2113 mg/100 grams
• Oyster 672 mg/100 grams
• Walnuts2542 mg per ounce, which amounts to 7 about
walnuts.
• Chia Seeds 4915 mg per ounce (28 grams).
• Flaxseeds 2338 mg per tablespoon of seeds, 7196 mg per
tablespoon of oil.
• Hemp Seeds 1000mg per tablespoon of seed
• Soybeen 1443 mg per 100 grams

20. OMEGA-3 AND BIOAVAILABILITY
• Very limited amount of Δ-6-desaturase in the digestive tract and
no more than 5% of ingested ALA can be converted in the final
metabolites EPA and DHA
• Δ-6-desaturase is more deficient in elderly, in diabetics,
hypertensives and in persosns who suffer from neurodegenerative
diseases.
• Direct supplementation with bioavailable omega 3 is more useful
• Dosage: 2-3 g/day

21. Antiinflammatory herbs
Zingiber officinalis(ginger roots)
-suppresses prostaglandin synthesis by inhibiting
cyclo-oxygenase-1 and cyclo-oxygenase-2
-suppresses leukotriene biosynthesis, by inhibiting 5-
lipoxygenase
-association with the Alpina galanga (Zingiberaceae
family), is able to inhibit the induction of several
genes involved in the inflammatory response.
-6-Shogaol performs an action against neuro-
inflammation in animal models of Parkinson's disease,
in LPS-induced inflammation and in transient
ischemies.
-6-shogaol can however play a role in the inhibition of
activation of glial cells and in reducing memory
deficits in animal models of dementia
Dosage: 300-600mg/day

22. Boswellia serrata
one of the ancient and most valued herbs in Ayurveda. “Gajabhakshya”, a Sanskrit name
sometimes used for Boswellia, suggests that elephants enjoy this herb as a part of their diet.
• Modern medicine and pharmacology strongly point out to its use as an:
• antiarthritic
• Antiinflammatory
• Antihyperlipidemic
• antiatherosclerotic (anticoronary plaque),
• Analgesic
• hepatoprotective
The anti-inflammatory effect is linked to its ability to inhibit certain
reactions, such as cyclo-oxygenase, just like the most classic anti-
inflammatories.
The anti-inflammatory mechanisms of Boswellia:- inhibition of 5-
lipoxygenase (5-LO). Boswellic acid selectively inhibit the 5-
lipoxygenase blocking the synthesis of leukotrienes: leukotriene B4
(LTB4) for acute inflammation and LTC4, LTD4, LTE4, for chronic
inflammation.
Dosage : 400-900mg/day
Ayurveda:
• antirheumatic (antiarthritis) activity
of gugguls-the gum-resins of trees
• Dysentery
• fevers (antipyretic),
• skin and blood diseases
• cardiovascular diseases
• asthma, cough,
• Astringent
• Diuretic
• hepatoprotective

23. Curcuma longa
-strong anti-inflammatory action
Suppresses genes that ratched up inflammation COX2gene
-detoxifying and immunostimulating agent
-the curcuminoids are also able to exert an antioxidant action,
with the blocking of free circulating radicals and the inhibition of
the formation of new ones.
-contributes to inhibiting angiogenesis, a process involved in the
development and proliferation of neoplastic cells . It is a potent
inhibitor of NF-kB and its inhibitory effect on NF-kB leads mainly
to a cellular apoptotic response.
-In addition, curcumin stimulates the expression of pro-apoptotic
Bax and inhibits the activation of the anti-apoptotic Mcl-1 and Bcl-
2, also altering the expression of the apoptotic mechanism
associated with NF-kB, p38 and p53 proteins.
poor bioavailability-recommended in association with bioperine or
liposomal curcumin
Dosage: 600-1000mg/day

24. Polygonum cuspidatum and resveratrol
resveratrol=natural polyphenol, able to exert an anti-
oxidant and anti-inflammatory activity.
Polygonum cuspidatum is one of the most naturally
occurring resveratrol plants.
• reduce significantly the production of NO and PGE2,
the expression of iNOS and COX-2 and the
production of ROS induced by the network of pro-
inflammatory cytokines.
• suppresses the mRNA expression of TNF-a and
promotes the mRNA expression of anti-
inflammatory molecules, interleukin-10
• promotes the expression of the BDNF important
molecule involved in the processes of neuronal
plasticity and in the response to the conditions of
hypoxic stress
Dosage: 100-300 mg /day

25. Evening Primrose oil-Oenothera biennis
rich in gammalinolenic acid
(GLA)
• GLA is metabolized to
dihomogammalinolenic acid
(DGLA), the immediate
precursor of prostaglandin
E1 (PGE1), an eicosanoid
with known anti-
inflammatory and
immunoregulatory
properties
• a competitive inhibitor of
PGE2 and LTs and thus
suppress inflammation
• Dosage 3-4g/day

27. Antiinflammatory diet
Abundant fruit and vegetables
Healthy fat: olive oil, avocado ,
nuts
Low glycemic load diet
Green tea
Turmeric
Ginger
Exercises
• Caloric restriction
decrease inflammation
• Intermitent fasting
decrease inflammation

28. Glycation
• Non-enzymatic reaction between reducing sugars, such as glucose,
and proteins, lipids or nucleic acids.
• Glycation has to be distinguished from glycosylation, which is an
enzymatic reaction.
• Since its first description by Maillard in 1912 and its involvement in
food browning during thermal processing by Hodge 50 years later, its
presence in living systems and involvement in various pathologies of
the human body, including aging and diabetes, have been an
intensive field of research.

29. Schematic presentation of the Maillard reaction.
Reactive carbonyl groups of a reducing sugar react with neutrophilic free amino groups of proteins to form a reversible Schiff
base. Through rearrangement a more stable Amadori product is formed. Dependent on the nature of these early glycation end
products, protein adducts or protein crosslinks are formed.

30. AGEs are a very heterogeneous group of molecules. Since the
discovery of the first glycated protein, glycated hemoglobin in
diabetes, numerous other AGEs have been detected.
The link between AGE and rAGE(receptor AGE) determines cellular activation and a process
of intracellular oxidative stress, which leads to the production of various types of
inflammatory cytokines, of various growth and transcription factors.
The rAGE-AGE bond, also tends to lead to self-amplification, more AGEs more rAGE develop,
with significant final tissue damage, linked to the generation and production of free
radicals and to the establishment of inflammatory processes.
No Known Positive Attributes of A.G.E.s

31. A.G.E.s Stimulate Multiple Inflammatory and Metabolic Pathways
AGEs RECEPTORS
Macrophages
Epithelial
Mesangial
Endothelium
AGE crosslinks
Multispecific AGE receptors
RAGE
AGE R-1
AGE R-2
AGE R-3-GALECTIN
MSRA
STIMULATION
OF
INFLAMMATION
AND
METABOLIC
PATHWAYS
Cytokines
(TNFa, IL-1, IL-6)
Extracellular Matrix
Production
(CTGF, TGFa)
Metabolic Pathway
Activation
(PKCa)
Gene Expression
(NFkB, ERK, 1, 2,
JNK)
Oxidative Stress
(NADPH Oxidase)
Adhesion Molecules
(VCAM-1, ICAM)
Growth Factors(TGFa,
CTGF, PDGF, VEGF, IGF-1

32. AGEs=Cofactor in aging process
Glycotoxins accumulate in our collagen and skin, cornea, brain and nervous
system, arteries and vital organs as we age.
Unfortunately, glycotoxins are highly resistant to the normal processes of protein
turnover and renewal that maintain the healthy tone of youthful body tissues
and organs.
It is a cofactor of age related deseases,particularly cardiovascular, kidney, eyes,
brain, skin diseases.
The damage caused by
glycation is irreversible

33. AGEs Sources
Modern Diet are linked to hight AGEs levels.
This is mostly due to popular methods of cooking that expose food to dry heat.
included barbecuing, grilling, roasting, baking, frying, sautéing, broiling, searing and toasting.
These cooking methods may make food taste, smell and look good, but they raise AGEs to
dangerous levels.
Dry heat causes AGE formation to increase by 10 to 100 times the levels in uncooked foods.
Foods highest in AGEs include meat (especially red meat), certain cheeses, fried eggs, butter,
cream cheese, margarine, mayonnaise, oils and nuts. Fried foods and highly processed
products also contain high levels.
So even if your diet appears reasonably healthy, you may consume an unhealthy amount of
harmful AGEs just because of the way your food is cooked.

34. How much is to much?
a high-AGE diet?
above 15,000 kilo units daily.
the amount of AGEs in common foods
1 fried egg: 1,240 kU/l
1 scrambled egg: 75 kU/l
2 ounces (57 grams) of toasted
bagel: 100 kU/l
2 ounces of fresh bagel: 60 kU/l
1 tablespoon of cream: 325 kU/l
¼ cup (59 ml) of whole milk: 3 kU/l
3 ounces of grilled chicken: 5,200 kU/l
3 ounces of poached chicken: 1,000 kU/l
3 ounces of french fries: 690 kU/l
3 ounces of baked potato: 70 kU/l
3 ounces (85 grams) of broiled
steak: 6,600 kU/l
3 ounces of braised beef: 2,200 kU/l

35. Clinical monitoring
AGE reader-indirect method

36. Hematochemical parameters
glycemia
glycated hemoglobin (Hb-glycated): is a much more useful parameter of glycemia in diagnosis and monitoring, as
it is an expression of long-term average blood sugar.
fructosamine: it is the product deriving from the glycation of albumin. Indicates the average glucose level in the 2-3
weeks prior to sampling; is not affected by food intake at the time of collection.
basal and postprandial insulinemia: the relationship between blood sugar and insulin has led to the
development of the HOMA index (Homeostasis Model Assessment) that highlights the risk or the peripheral resistance
to insulin, both in the individual with normal body mass index and in the obese and in the prediabetic.

37. Glycation Treatment
AGEs restricted diet
LGL diet
Exercises
Nutraceuticals
Carnosine
Pyridoxamine
Zingiber officinalis
Alfa lipoic acid
Aerva Lanata

38. Carnosine
Dipeptide molecule, made up of the amino acids beta-alanine and histidine.
It is highly concentrated in muscles and brain tissues.
• antioxidant properties. ROS scavenger
• acts as an antiglycating agent,
• suppresses glycation by chelating metal ions, reducing possible damage to the DNA
itself.
• it also protects the other protein molecules from the toxic effects related to the
production of AGE, acting almost as a "sacrificial site" (glycating themselves) and, by
binding to the glycated proteins, it favors its elimination.
• Intracellular buffer-delay senescence in fibroblasts
• is also able to inhibit the aggregation of beta-amyloid peptides in Alzheimer's disease
and to counteract the harmful effects linked to senescence of alpha-synuclein in
Parkinson’s disease
• Useful in skin aging treatment
• Dosage: 1000mg/day
Properties

39. PM(Pyridoxamine)
• Pyridoxamine(PM)is one of 3 naturally occuring vitamers of
Vitamin B6. The other B6 vitamers are pyridoxal (PL) and
pyridoxine (PN), the latter being the form primarily used in
supplements currently produced by suppliers. PN is found mainly
in plant sources and PM and PL are found almost exclusively in
animal sources.
• Various forms of vitamin B6 have been used for decades for
dietary supplemental and medical therapeutic purposes.
Although PM has not itself been used as much as other forms, all
indications are that its safety and toxicity profile should be at
least as good as the other forms.
• In chronic use at a dose under 100 mg daily, there are no reports
of any negative effects from the use of vitamin B6.
• Powerful inhibitor og AGE and ALE(advanced lipoxidation end
products)

40. Alpha-Lipoic Acid
 mitochondrial fatty acid highly involved in energy metabolism. It is synthesized in the body
and can be consumed through eating meat.
 in supplement form, it has shown benefit for various forms of oxidation and inflammation.
These effects protect against heart diseases, liver diseases, diabetes, and neurological
decline associated with aging.
 ALA is a potent anti-oxidant compound. It works with mitochondria and the body's natural
anti-oxidant defenses.
 ALA is also seen as an anti-aging compound since it can reverse some of the oxidant damage
related to the effects of aging.
 Appears to reduce biomarkers of oxidation
 Appears to be a slight reducing effect on HbA1c
 Appears to reduce biomarkers of lipid peroxidation (MDA mostly)
 DOSAGE: 450-600mg/day

41. Methylation, gene expression and aging
What is methylation?
 a process by which methyl groups are added to the DNA molecule.
 can change the activity of a DNA segment without changing the sequence.
 located in a gene promoter, DNA methylation typically acts to repress
gene transcription.
 DNA methylation is essential for normal development and is associated with a
number of key processes including genomic imprinting, X-chromosome
inactivation, repression of transposable elements, aging and carcinogenesis,
DNA repair
 Two of DNA's four bases, cytosine and adenine, can be methylated.
 Methylation requires an abudant supply of methyl donors( Methionine, Choline,
SAMe) and of methylating factors: Vit B6, B12, Folic acid, Zn.
 When methylation is deficient good genes get turned OFF and bad genes get
turned ON.
Hight plasmatic levels of homocysteine are a signal of impaired methylation

42. Age-specific DNA methylation changes and age-related diseases
 DNA methylation patterns change over time →role in age-related diseases.
 The number one age-related disease is cancer and indeed one of the best
predictors of tumors is the age of patients
 In most cancer types, global DNA hypomethylation can be observed →potential
causal factor in reducing genome stability and increasing chromosomal
aberrations.
 For some genes, site-specific hypermethylation of promoter regions are found in
both aging and tumorigenesis, which makes them bona fide candidates for
increased cancer susceptibility.
 Examples: insulin-like growth factor-II (IGFII) hypermethylated in cancer 1
(HIC1), caspase-8 (CASP8), glutathione S-transferase pi (GSTP1), suppressor of
cytokine signaling 1 (SOCS1), RAS association domain family 1A (RASSF1A),
p16/CDKN2A, adenomatosis polyposis coli (APC) and estrogen receptor 1 (ESR1)

43. Age-specific DNA methylation changes and age-related diseases
 inflammatory diseases show an age-related increase of DNA methylation. Inflammation is
often associated with DNA hypermethylation of specific genes as initially reported for
ulcerative colitis.
 the gradual decrease of methylation in promoter fragments of the (TNFα) gene, which is
important for inflammatory reactions, could be a main factor in the onset of chronic
inflammatory conditions with older age
 Alzheimer’s disease, amyloid precursor protein (APP) gene showed gradual
hypomethylation in the promoter. Age-dependent changes of methylation for the
presenilin 1 (PS1) gene, which is essential for the formation of the γ-secretase complex,
also imply gene expression changes.
 reduction of DNA methylation decrease the resting metabolic rate.
 hypomethylation caused by increased levels of homocysteine and S-
adenosylhomocysteine (SAH), leads to inhibition of cellular methylation reactions.
 it is currently assumed that regular intake of nutrients involved in the metabolism of the
methyl group, like folic acid or vitamin B12, can slow down the gradual hypomethylation
observed during the aging process.
 However, the relationship between folate status and DNA methylation levels is complex
and is likely influenced by folate availability

44. Homocysteine
 nonprotein sulfur-containing amino acid
 Derived from metabolic demethylation of dietary methionine
 Present in plasma in more forms:
 <1% free reduced
 70-80 % bound to proteins
 20-30 % free oxidized

45. FACTORS THAT INCREASE PLASMA HOMOCYSTEINE LEVELS

46. Lifestyle on tHcy levels in the Hordaland
Homocysteine Study. JAMA 1995.
 Moderate alcohol consumption
 Chronic high alcohol consumption
 Beer consumption
 More than 4 coffee /day
 Cigarette smoking
 Diet with less than
B6 1.2 mg/day
Folic acid 0.5 mg/day
B12 2.4 ug/day

47. Homocysteine and disease-CAD
CAD- hyperhomocysteine negatively affect endothelial cell function
A Predictive factor for hypertension-circulating homocysteine is related
to increased arterial stiffness in prehypertensive patients

48. HOMOCYSTEINE HAS ATHERO-THROMBOTIC PROPERTIES
 Intimal thickening
 Elastic lamina disruption
 Smooth muscle hypertrophy
 Marked platelet accumulation
 Platelet-enriched occlusive thrombi

49. Hyperhomocysteinemia as a Risk Factor for Atherosclerosis
 Nitric oxide (NO) not only relaxes the arteries, but also prevents plaque
formation
 A high concentration of homocysteine blocks EDRF( Endothelial derived
relaxing factor) thus initiating the arteriosclerotic process.
 Furthermore the homocysteine irritates the muscle cells of the arteries
causing a proliferation of the arteriosclerotic process.

50. Physiological conditions
NO+Homocysteine
S-NITROSO-HOMOCYSTEINE
Antiplatelets activity
vasodilatation
Auto-oxidation
ROS

51. Pathological condition
NO+ Homocysteine
S-NITROSO-HOMOCYSTEINE
Auto-oxidation
ROS
Antiplatelets activity
vasodilatation

52.  Hyperhomocysteinemia induces inhibition of
dimethylarginine-dimethylaminohydrolases
(DDAH), the enzymes that normally degrade
asymmetric dimethylarginine (ADMA).
 ADMA levels are increased in
hyperhomocysteinemia
 ADMA inhibits basal and stimulated
endothelium-dependent relaxation in cerebral
blood vessels; moreover it inhibits production
of NO by endothelial NOS (eNOS) and also may
promote eNOS uncoupling.

53. The relationship has been investigated between
homocysteine and cognitive decline in 1,241
subjects aged 61 to 73 years, followed up over 4
years
Cross-sectional analyses showed that higher
concentrations of homocysteine were
significantly related to poorer performances at
all neuropsychological tests. Longitudinal
analyses confirmed this finding.
The odds of cognitive decline
was 2.8-fold ( p
< 0.05) higher in subjects
with homocysteine levels
above 15mol/L compared
with those with
homocysteine levels below
10mol/L.

54. Homocysteine level and Alzheimer’s disease eine
Temporal medial lobe atrophy and
HyperHcy in AD
Hyppocampal width decreases with age
In 156 subjects with non clinical memory problems,
hippocampal width (MRI), decresed significantly with
increasing Hcy levels( no effect of age)
Hyper Hcy damage Hippocampus
Williams J h et al., Minimal Hyppocampal width relates to plasma Hcy in community dwelling older
people. Age Ageing, 2002:31 440-444.

55. Studies of newly diagnosed dementia are
required in order to establish whether the
elevated homocysteine levels precede
the onset of dementia or result from
dementiarelated nutritional and vitamin
deficiencies.
A total of 1092 subjects without dementia
(667 women and 425 men; mean age, 76
years) from the Framingham Study
constituted our study sample.
Mean Follow up: 8 yrs (1-11).
111 developed dementia (83 AD).

56. An increased plasma Hcy level is a strong,
independent risk factor for the development
of dementia and Alzheimer’s disease.
With a plasma level > 14
µmol/l, the risk nearly doubles.

57. The correlation between serum folate and the severity of
atrophy of the neocortex was 20.40 (P = 0.03) (N.30) .
Among a subset of 15 participants with significant
numbers of Alzheimer disease lesions in the neocortex, the
correlation between folate and atrophy was 20.80 (P =
0.0006).
Conclusion.
Low serum folate is strongly associated with
atrophy of the cerebral cortex.

58. Conclusion: Elevated tHcy levels are associated
with decreased cognitive performance in non
demented elderly people, and the relation was most
marked for psychomotor speed.
This association was independent of structural brain
changes (atrophy; WMH; lacune) on MRI.

59.  Individuals with higher plasma
homocysteine levels had, on average,
more cortical atrophy and more
hippocampal atrophy
No association was observed between
plasma homocysteine levels and
amygdalar atrophy
• Results support the hypothesis that
higher plasma homocysteine levels are
associated with more atrophy of the
hippocampus and cortical regions in
elderly at risk of Alzheimer's disease.

60. Methylation. Clinical monitoring
Genetic aspects Plasmatic homocysteine
concentration
Optimum ranges: 5-
10µmol/L
Other plamsmatic
parameters
Folic acid
B12
B6
Zn r

61. MTHFR, C677T polymorphysm and Homocysteine level
MTHFR plays a critical role in homocysteine
metabolism by catalyzing the conversion of
5, 10 methylenetetrahydrofolate to
5-methyltetrahydrofolate, the predominant
circulatory form of folate and the methyl-group
donor in the B12-dependent remethylation of
homocysteine to methionine
Two common polymorphisms of MTHFR gene, the
thermolabile C677T and A1298C polymorphism
may contribute to hyperhomocysteinemia

62. Lowering plasma Homocysteine
Mechanisms of Homocysteine Detoxification
• A potent remethylation agent is TMG, which stands for trimethylglycine.
The tri means there are three methyl groups on each glycine molecule
that can be transferred to homocysteine to transform (remethylate) it into
methionine and SAMe.
• The remethylation (or detoxification) of homocysteine requires the
following minimum factors: (1) folic acid, (2) vitamin B12, (3) zinc, and
(4) TMG.

63. Basic Pro-methylation Protocol
Folic Acid 1600mcg
B6 100mg
B12 500mcg
TMG 750mg

64. ADVANCED PRO-METHYLATION
PROTOCOL
• Folic acid,up to 10000 mcg a day.
• Vitamin B12,up to 5000 mcg a day.
• Vitamin B6,up to 300 mg a day.
• Pyridoxal-5-phosphate(P5P),up to 50 mg.
• Zinc, 30 to 90 mg a day.
• Choline, up to 5000 mg a day.
• Trimethylglycine, up to 6000 mg a day.
• SAMe,up to 800 mg a day.

65. Oxidative stress

66. Reactive oxygen species (ROS) are produced by living organisms as a result of
normal cellular metabolism and environmental factors, such as air pollutants or
cigarette smoke.
ROS are highly reactive molecules and can damage cell structures such as
carbohydrates, nucleic acids, lipids, and proteins and alter their functions.
Aerobic organisms have integrated antioxidant systems, which include enzymatic
and nonenzymatic antioxidants that are usually effective in blocking harmful
effects of ROS.
However, in pathological conditions, the antioxidant systems can be overwhelmed

67. Oxidative stress contributes to many pathological conditions and diseases

68. Oxidants
Endogenous Sources of ROS
 ROS are produced from molecular oxygen as a result of normal cellular metabolism.
 ROS can be divided into 2 groups: free radicals and nonradicals.
 Molecules containing one or more unpaired electrons and thus giving reactivity to
the molecule are called free radicals.
 When 2 free radicals share their unpaired electrons, nonradical forms are created.
The 3 major ROS that are of physiological significance are
superoxide anion (O2
−.), hydroxyl radical (•OH), and
hydrogen peroxide (H2O2).

69. Exogenous Source of Oxidants

70. Antioxidants
The human body is equipped with a variety of antioxidants that serve to
counterbalance the effect of oxidants. For all practical purposes, these
can be divided into 2 categories:
enzymatic and nonenzymatic
enzymatic

71. Nonenzymatic Scavenger of Antioxidant Defenses
Scavenger against various
radicals
(HO •, ROO • and O2)
Reducing the tocopheryl
radical regenerates vitamin E

72. Glutathione
GSH is highly abundant in all cell
compartments and is the major soluble
antioxidant.
GSH detoxifies hydrogen peroxide and lipid
peroxides via action of GSH-Px.
GSH donates its electron to H2O2 to reduce it
into H2O and O2.
Reduced glutathione donates protons to
membrane lipids and protects them from
oxidant attacks.
GSH/GSSG ratio
is a major biomarker of oxidative stress

73. Vitamin E (α-Tocopherol)
 Lipid-soluble vitamin E is concentrated in the hydrophobic
interior site of cell membrane and is the principal defense
against oxidant-induced membrane injury.
 Vitamin E donates electron to peroxyl radical, which is
produced during lipid peroxidation.
 α-Tocopherol is the most active form of vitamin E and the
major membrane-bound antioxidant in cell.
 Vitamin E triggers apoptosis of cancer cells and inhibits
free radical formations.

74. Carotenoids (β-Carotene)
Carotenoids are pigments found in plants.
Primarily, β-carotene has been found to react with
peroxyl (ROO•), hydroxyl (•OH), and superoxide (O2
−.)
radicals.
Carotenoids show their antioxidant effects in low
oxygen partial pressure but may have pro-oxidant
effects at higher oxygen concentrations.
Both carotenoids and retinoic acids (RAs) are capable
of regulating transcription factors.
β-Carotene inhibits the oxidant-induced NF-κB
activation and interleukin (IL)-6 and TNFα production.
Carotenoids also affect apoptosis of cells.
Antiproliferative effects of RA have been shown in
several studies. This effect of RA is mediated mainly by
retinoic acid receptors and vary among cell types.
In mammary carcinoma cells, retinoic acid receptor
was shown to trigger growth inhibition by inducing cell
cycle arrest, apoptosis, or both
Βcaroten molecule 3D image

75. Effects of Oxidative Stress on DNA
ROS can lead to DNA modifications in several ways:
Degradation of basis
Single or double strand DNA break
purine, pyrimidine or sugar-bound modifications
mutations, deletions or translocations
cross-linking with proteins.
Most of these DNA modifications are highly relevant to carcinogenesis, aging, and
neurodegenerative, cardiovascular, and autoimmune diseases.
Tobacco smoke, redox metals, and nonredox metals, such as iron, cadmium, chrome, and
arsenic, are also involved in carcinogenesis and aging by generating free radicals or binding
with thiol groups.
Formation of 8-OH-G is the best-known DNA damage occurring via oxidative stress

76. Clinical monitoring
d-ROMs test
BAT test
(biological
antioxidant
potential)
 The d-ROMs test essentially determines the concentration of hydroperoxides (ROOH) in the
blood
 can be performed on samples of whole blood (generally finger prick capillary blood)
 the units of measurement for the test is expressed in U. CARR.
 normal range 250-300 uCARR
 One U. CARR is equal to 0.08 mg/dL of a solution of hydrogen peroxide.

77. BAT test (biological antioxidant potential)
The test can be performed on venous serum
it examines the blood concentration of
antioxidants as agents that can reduce iron
from the ferric (Fe3+) to ferrous (Fe2+) form

78. • Measurement of Urinary 8-OHdG
• Measurement of F2-isoprostane
• Measurement of Malondialdehyde
Other test

79. Antioxidant therapy
Vitamine C
Vitamine E
Glutamine
Combined antioxidants (selenium, β-
carotene, vitamin C, vitamin E and
methionine)
Glutathione precursors [S-adenosyl
methionine (SAMe)]
Coenzyme Q10
Pycnogenol
Alfa lipoic acid
Resveratrol
Quercetin
Hesperedin

80. Conclusions
It is not possible to stop aging but we can slow
it down
Diet plays an important role and the right
choice of nutrients can help in prevention of
illness and increase the quality of life.
There is great interest in anti-aging substances
derived from food, nutraceuticals, such as
antioxidants, herbs, omega-3 fatty acids. These
substances have beneficial effect on digestive
and immune systems, and modulate
inflammatory and degenerative processes in
the body.
Exercise are very important too in healthy
aging

81. Age is an issue of mind over
matter.
If you don’t mind, it doesn’t
matter 
Vranceanu.Maria@umfcluj.ro
Thank you!

82. References
1. Alic N., Partridge L. Death and dessert: nutrient signaling pathways and ageing. Curr Opin Cell Biol. 2011;23:738–7436.
2. De Luca d'Alessandro, Bonacci S., Giraldi G. Aging populations: the health and quality of life of the elderly. Clin Ter. 2011;162:e13–e18.
3. De Martinis M., Franceschi C., Monti D., Ginaldi L. Inflamm-ageing and lifelong antigenic load as major determinants of ageing rate and
longevity. FEBS Letters. 2005;579(10):2035–2039. doi: 10.1016/j.febslet.2005.02.055.
4. Fontana L., Partridge L., Longo V.D. Extending healthy life span—from yeast to humans. Science. 2010;328:321–3264.
5. Franceschi C. Cell proliferation, cell death and aging. Aging Milano. 1989;1(1):3–15.
6. Franceschi C., Bonafè M., Valensin S., et al. Inflamm-aging. An evolutionary perspective on immunosenescence. Annals of the New York Academy
of Sciences. 2000;908:244–254.
7. Franceschi C., Monti D., Sansoni P., Cossarizza A. The immunology of exceptional individuals: the lesson of centenarians. Immunology
Today. 1995;16(1):12–16. doi: 10.1016/0167-5699(95)80064-6.
8. Giunta B., Fernandez F., Nikolic W. V., et al. Inflammaging as a prodrome to Alzheimer's disease. Journal of Neuroinflammation. 2008;5, article
51 doi: 10.1186/1742-2094-5-51.
9. Haigis M.C., Yankner B.A. The aging stress response. Mol Cell. 2010;40:333–344.
10. Harman D. The free radical theory of aging. Antioxid Redox Signal. 2003;5:557–61.
11. Jones PA, Baylin SB. The epigenomics of cancer. Cell 2007; 128: 683-92. [4] Trojer P, Reinberg D. Histone, lysine, demethylases and their impact on
epigenetics. Cell 2006; 125: 213-7. [5] Scriver CR. Nutrient gene interactions: the gene is not the disease and viceversa. Am J Clin Nutr 1988; 48:
1505-9.
12. Michikawa Y, Mazzucchelli F, Bresolin N, Scarlato G, Attardi G. Aging-dependent large accumulation of point mutations in the human mtDNA control
region for replication. Science. 1999;286:774–9. doi: 10.1126/science.286.5440.774.
13. Prakash D., Gupta C. Role of antioxidant polyphenols in nutraceuticals and human health. In: Prakash D., Sharma G., editors. Phytochemicals of
Nutraceutical Importance. CABI International Publishers; Wallingford: 2014. pp. 208–228.
14. Rodenhiser D, Mann M. Epigenetics and human disease: translating basic biology in to clinical applications. Can Med Assoc J 2006; 174: 341-8.
15. Salminen A., Kauppinen A., Kaarniranta K. Phytochemicals suppress nuclear factor-kappaB signaling: impact on health span and the aging
process. Curr Opin Clin Nutr Metab Care. 2012;15:23–28.
16. Salmon A.B., Richardson A., Pérez V.I. Update on the oxidative stress theory of aging: does oxidative stress play a role in aging or healthy
aging? Free Radic Biol Med. 2010;48:642–655.
17. Thorpe SR, Baynes JW. Maillard reaction products in tissue proteins: new products and new perspectives. Amino Acids. 2003;25:275–81. doi:
10.1007/s00726-003-0017-9.