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Vegetarian Diet Affects Genes of Oxidative Metabolism and Collagen Synthesis

  • Ludwig Boltzmann-Cluster Oncology (LB-CO) | Medical University Vienna

Abstract and Figures

A vegetarian diet is known to prevent a series of diseases but may influence the balance of carbohydrate and fat metabolism as well as collagen synthesis. This study compares expression patterns of relevant genes in oral mucosa of omnivores and vegetarians. Quantitative reverse transcriptase polymerase chain reaction was applied for analysis of mRNA levels from carnitine transporter OCTN2, hepatic CPT1A and nonhepatic CPT1B isoforms of carnitine palmitoyltransferase and collagen (CCOL2A1) in oral mucosa. Compared with volunteers with traditional eating habits, carbohydrate consumption was significantly higher (+22%) in vegetarians. This was associated with a significant stimulation of CPT1A (+50%) and OCTN2 (+10%) and a lowered collagen synthesis (-10%). These novel findings provide further insight into the association of a changed fat metabolism and reduced collagen synthesis in vegetarians, which could also play a role in the aging process.
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Original Paper
Ann Nutr Metab 2008;53:29–32
DO I: 10 .1159/000152 871
Vegetarian Diet Affects Genes of
Oxidative Metabolism and Collagen
Heidrun Karlic a Daniela Schuster a, c Franz Varga b Gerhard Klindert d
Alexander Lapin d Alexander Haslberger a, c Michael Handschur
a Ludwig Boltzmann Institute for Leukemia Research and Hematology,
b Ludwig Boltzmann Institute of Osteology
at the Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 4th Medical Department, Hanusch Hospital,
c Department of Nutritional Sciences, University of Vienna, and
d Sozialmedizinisches Zentrum Sophienspital der
Stadt Wien, Vienna , Austria
It is well established that a vegetarian diet may lower
the risk of cancer (e.g., colon cancer) and age-associated
diseases [for a review, see ref.
1 ]. A vegetarian diet may
also shift the macronutrient balance (fat and proteins vs.
carbohydrate) in favor of carbohydrates
[2, 3] . As a high-
er protein intake is known to optimize gains in muscle
[4] and stimulates fat metabolism [5] , a carbohy-
drate-rich diet could stimulate a downregulation of key
enzymes of fat metabolism.
There is evidence t hat the transcription of about 25,000
genes is regulated by nutrition, exercise and hormones
[6–8] . However, the transcription of 3 fat metabolism-
associated genes, OCTN2 (organic cation transporter),
CPT1A and CPT1B (hepatic and muscular isoform of
carnitine palmitoyltransferase), is regulated in response
to diet, exercise and aging
[9, 10] .
In addition, nutritional factors are associated with
buildup of musculoskeletal mass [for a review, see e.g. ref.
11 ]. As there is a close correlation of protein synthesis
with mRNA expression for collagen (CCOL2A1)
[10 , 12] ,
the latter provides a suitable model for evaluating effects
on the expression of relevant proteins.
Our assumption that many metabolism-associated
genes are regulated in a systemic manner results from a
Key Words
Vegetarian diet mRNA expression Metabolism
Collagen Omnivores
Background/Aim: A vegetarian diet is known to prevent a
series of diseases but may influence the balance of carbohy-
drate and fat metabolism as well as collagen synthesis. This
study compares expression patterns of relevant genes in
oral mucosa of omnivores and vegetarians. Methods: Quan-
titative reverse transcriptase polymerase chain reaction was
applied for analysis of mRNA levels from carnitine transport-
er OCTN2, hepatic CPT1A and nonhepatic CPT1B isoforms of
carnitine palmitoyltransferase and collagen (CCOL2A1) in
oral mucosa. Results: Compared with volunteers with tradi-
tional eating habits, carbohydrate consumption was signifi-
ca nt ly h ig her (+22 %) i n ve ge ta ria ns . T his wa s as so cia ted w it h
a significant stimulation of CPT1A (+50%) and OCTN2 (+10%)
and a lowered collagen synthesis (–10%). Conclusion: These
novel findings provide further insight into the association of
a changed fat metabolism and reduced collagen synthesis
in vegetarians, which could also play a role in the aging pro-
cess. Copyr ight © 2008 S. Karger AG, B asel
Received: July 5, 20 07
Accepted after revision: June 17, 2008
Publis hed online: Septemb er 5, 2008
H. Karlic
Ludwig Boltzmann Institute for Leu kemia Research and Hematology
Hanusch Hospital, Heinrich Colli nstrasse 30 , AT–1140 Vienna (Austri a)
Tel. +43 699 192 4 1457, Fax +43 699 1914 3214
© 200 8 S. Karger AG, Basel
Accessible online at:
Karlic /Schuster /Varga /Klindert /Lapin /
Ann Nutr Metab 2008;53:29–32
larger comparative study using white blood cells and
[13] as well as from preliminary experiments for
this study comparing white blood cells and oral mucosa.
Thus, oral mucosa obtained from mouth swabs was cho-
se n to eva luate po ssible differences in gene reg ulation b e-
tween probands with traditional eating habits and vege-
Materials and Methods
Study Population
A group of 116 volunteers participated in this study: 86 omni-
vores (43 females/43 males, mean age 24 years, range 18–31) were
compared with 30 vegetarians (15 females/15 males, mean age 23
years, ra nge 19–26). Nutritiona l questionnaires bas ed on a previous
European study
[14] were given to all participants. Only subjects
with a normal body mass index (BMI 18.5–24.9) were included.
N o n i n v a s i v e S a m p l i n g
Buccal cells were collected on sterile cotton sticks by twirling
at least 10 times on each of the inner cheeks. Then the cells were
released into 1 ml storage buffer for nucleic acid preparation
(commercially obtained from Roche Diagnostics).
Analysis of mRNA Expression
Isolation of mRNA and preparation of cDNA was carried out
according to standard procedures. Quantitative reverse tran-
scriptase polymerase chain reaction (RTQ-PCR) was carried out
using a LightCycler
TM System (Roche) or a Rotorgene 6000, which
allows amplification and detection (by fluorescence) in the same
tube, using a kinetic approach. Quantitative PCR was done using
a real-time PCR system (RTQ-PCR) which was also applied for
analysis of mRNA expression using commercially available Taq-
Man primers and probes (Applied Biosystems). As a control, to
confirm the intra-indiv idual stabilit y of gene expression, samples
of 1 volunteer obtaine d on 5 dif ferent days were ana lyzed showing
identical gene expression patterns (SD ! 1%).
S t a t i s t i c s
Evaluation of results was done using the StatView 5.01 statis-
tics program using ANOVA to find out significant differences
between the groups. All tests were done double sided at a signifi-
cance level of 5% (p ! 0.05).
R e s u l t s
Results of Nutritional Questionnaires
Evaluation of nutritional questionnaires indicated
that nutritional habits of vegetarians differed mainly
with respect to the refusal of meat consumption and a
relatively higher intake of milk products (610 meals/
week) in the majority of vegetarians (55%, SD 10%) com-
pared with 3–6 meals/week in the majority of the om-
nivorous group (60%, SD 15%). In addition, one fish meal
was reported in the omnivorous group (85%, SD 17%) but
not in vegetarians. Thirty percent (SD 10%) of volunteers
from each group reported an intake of supplements (vi-
tamins, 2–3 times/week).
Expression of Fat Metabolism-Associated Genes
Transcript levels of OCTN2, CPT1A and CPT1B are
shown in figure 1 .
The mean expression rate of organic cation transport-
er OCTN2 in buccal mucosa from omnivores was 75%
(SD 7%) of the standard gene (glucose 6-phosphatase,
G6PD). OCTN2 expression was significantly higher in
vegetarians (84% of G6PD, SD 3%; p ^ 0.05).
In omni vore s, t he mean ex pression r ate of CPT1A was
88% of G6PD (SD 8%), which was significantly higher in
vegetarians (153% of G6PD, SD 8%; p ^ 0.001). In omni-
vores, the mean expression rate of CPT1B was 88% of
G6PD (SD 8%) compared with 78% of G6PD (SD 8%; p !
0.05) in vegetarians. Interestingly, there was a 1:
1 ratio
between CPT1A and CPT1B in omnivores but a 2:
1 ratio
between CPT1A and CPT1B in vegetarians.
Relative mRNA expression
Fig. 1. Gene expression of fat metabolism-associated genes. Com-
pared with omnivores, the gene expression rate (percentage of
mRNA in relation to the standard gene G6PD) of OCTN2 (re-
sponsible for carnitine uptake) is significantly higher in vegetar-
ians (
a p ^ 0.05). Considering mitochondrial carnitine palmito-
yltransferases, CPT1A is significantly upregulated (
b p ^ 0.001)
and CPT1B is significantly downregulated in vegetarians (
c p ^
0.05). There was a 1:
1 ratio between CPT1A and CPT1B in omni-
vores and a 2:
1 ratio between CPT1A and CPT1B in vegetarians.
Vegetarians and Genes of Oxidative
Metabolism and Collagen Synthesis
Ann Nutr Metab 2008;53:29–32
Synthesis of Collagen (CCOL2A1)
As shown in figure 2 , relative mRNA levels of
CCOL2A1 in oral mucosa were higher than the standard
gene G6PD in all cases. However, the expression was re-
duced by 10% in vegetarians (140% of G6PD, SD 5%) as
compared with omnivores (157% of G6PD, SD 5%; p ^
Our new model system uses noninvasive sampling of
oral mucosa for the analysis of mRNA synthesis of me-
tabolism-associated genes which are known to be regu-
lated in a systemic manner. Taken together, we could
show that a vegetarian diet has a significant impact on
genes regulating essential features of carnitine metabo-
lism, which are also affected in a series of diseases rang-
ing from diabetes to cancer
[9] .
The stimulation of carnitine uptake as evidenced by
an elevated OCTN2 expression which was observed in
vegetarians participating in this study may compensate
lower carnitine levels which are usually sufficient not to
lead to any evident metabolic disturbances. An existing
systemic carnitine deficiency resulting for example from
inherited mutations of the OCTN2 gene is dramatically
worsened by a strictly vegetarian diet
[15] .
Intracellular carnitine is also essential for the trans-
port of fatty acids into mitochondria as mediated by car-
nitine palmitoyltransferases
[16] . This could support the
interpretation of the difference in the CPT1A/CPT1B re-
lation between omnivores (1:
1) and vegetarians (2: 1), re-
sulting from an upregulation of CPT1A and a downregu-
lation of CPT1B, in the sense that carbohydrate metabo-
lism is favored in vegetarians, whereas in omnivores,
there is a more balanced relation between carbohydrate
and fat metabolism.
The stimulatory effect of protein supplementation on
fat metabolism supports a plethora of previous data [for
a review, see e.g. ref.
11, 17 ] documenting the importance
of dietary protein for the maintenance of musculoskeletal
Relative mRNA levels
Omnivores Vegetarians
Fig. 2. G ene ex pres si on of co ll ag en. As co mpa re d w it h om ni vo re s,
the gene expression rate (percentage of mRNA in relation to the
standa rd gene G6PD) of collagen (CCOL2A1) is signif icantly low-
er in vegetarians ( * p ^ 0.05).
Tab le 1. Primers for mRNA analyses
Gene Sequence Reference
Karlic et al. [9]
Razeghi et al.
[20], 2001
Razeghi et al.
[20], 2001
This study
Hochhaus et al.
[21], 1996
Karlic /Schuster /Varga /Klindert /Lapin /
Ann Nutr Metab 2008;53:29–32
mass and confirms our observation indicating reduced
synthesis of collagen (CCOL2A1) in vegetarians. In addi-
tion, an increased risk of osteoporosis, which is also
known to be associated with a lowered collagen synthesis,
has been associated in vegetarians with a low cobalamin
(vitamin B
12 ) status. Cobalamin is an essential vitamin
for DNA synthesis and is primarily present in a protein-
bound form in foods of animal origin
[18] . Additional
evidence for collagen disorders resulting from a low-pro-
tein diet is given for a series of cutaneous manifestations
[19] .
In conclusion, our data show that a vegetarian lifestyle
has an impact on fat metabolism causing a remarkable
stimulation of carnitine uptake and a reduction in colla-
gen synthesis-associated genes.
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... These studies have found that the risk of major chronic diseases such as obesity, heart disease, certain cancer, some chronic degenerative disease, stroke, and type-2 diabetes may be reduced by adopting a completely plant-based diet (Craig, 2010;Key, Appleby, Davey, Allen, & Spencer, 2003;(American Dietetic Association, 2003;Craig & Mangels, 2009) Key, Appleby, & Rosell, 2006;Le & Sabate, 2014;Mishra, Xu, Agarwal, Gonzales, Levin, & Barnard, 2013;Tonstad, Stewart, Oda, Batech, Herring, & Fraser, 2013). On the other hand, other studies have shown that restrictive and monotonous vegetarian diets bring a risk of nutritional deficits (Appleby, Thorogood, Mann, & Key, 1999;Craig, 2009;Fraser, 2009;Karlic et al., 2008;McEvoy, Temple, & Woodside, 2012). Based on this research, there are two possible effects of a vegetarian diet on health. ...
Research on the relationship between vegetarianism and subjective well-being (SWB) has produced inconsistent results, which may partly be due to small sample sizes and divergent operationalizations of well-being. For these reasons, the present study aimed to thoroughly examine this association in two large representative samples from Germany (Study 1: N = 12,905, including 665 vegetarians) and Australia (Study 2: N = 15,532, including 383 vegetarians) using a consensual conceptualization of SWB (composed of an affective component, i.e., positive and negative affect, and a cognitive component, i.e., life satisfaction). Results of t-tests showed that vegetarians reported slightly higher scores in negative affect (Study 1 and 2), but also slightly higher levels of satisfaction with health (Study 1 and 2) and life satisfaction (Study 1) compared to meat eaters. No differences emerged regarding positive affect in either study. These differences between vegetarians and meat-eaters in some components of SWB, although significant due to the large sample sizes, are small at best (d around .15). Because sex, age, and education were associated with diet type and SWB, analyses controlling for socio-demographic variables were also conducted. In these ANCOVAS, the effect of diet emerged only for one out of the five formerly significant comparisons (negative affect in Study 1, d = .09) while the differences between vegetarians and meat eaters in both satisfaction with health and life, as well as the effect on negative affect in Study 2, all became non-significant (d around .05). Taken together, the very small effects found in the t-tests for some components of SWB seem to be due to socio-demographic variables, meaning that the true effects of diet on SWB are non-existent or negligible.
... Collagen genes have not commonly been examined in the context of dietary change. One such rare study revealed that vegetarian diet decreases COL2 A1 gene expression in the mucosa, suggesting that diets affect the collagen metabolic pathway [23] . Interestingly, the weight loss achieved by VLCD in this study was mostly due to loss of lean body mass (LBM) rather than a depletion of fat stores. ...
Background: In bariatric surgery, preoperative very low-calorie diets (VLCD) may better meet the technical demands of surgery by shrinking the liver. However, diets may affect tissue healing and influence bowel anastomosis in an as-yet-undefined manner. Objective: This randomized controlled trial aimed to examine the effect on collagen deposition in wounds in patients on a 4-week VLCD before laparoscopic gastric bypass. Setting: University hospital. Methods: The trial was undertaken in patients undergoing laparoscopic Roux-en-Y gastric bypass, with a control group (n = 10) on normal diet and an intervention group (n = 10) on VLCD (800 kcal) for 4 weeks. The primary outcome measured was expression of collagen I and III in skin wounds, with biopsies taken before and after the diet and 7 days postoperatively as a surrogate of anastomotic healing. Secondary outcome measures included liver volume and fibrosis score, body composition, operating time, blood loss, hospital stay, and complications. Results: Patients in both groups were similar in age, sex, body mass index (53.4 versus 52.8 kg/m2), co-morbidities, liver volume, and body composition. Expression of mature collagen type I was significantly decreased in diet patients compared with controls after 4 weeks of diet and 7 days after surgery. This was significant decrease in liver volume (23% versus 2%, P = .03) but no difference in operating times (129 versus 139 min, P = .16), blood loss, length of stay, or incidence of complications. Conclusions: Preoperative diets shrink liver volume and decrease expression of mature collagen in wounds after surgery. Whether the latter has a detrimental effect on clinical outcomes requires further evaluation.
... Improvement after fillers has been related to mechanical stress, inducing collagen synthesis through fibroblasts activation. Collagen synthesis can also be modulated by diet, with vegetarian diet being associated with reducing this synthesis (Karlic et al., 2008). However, the longevity of fillers has been recently related to local content of adipose-derived stem cells as well as on filler implantation ability to proliferate and to differentiate which can be related to the metabolic status of different subjects (Kruglikov & Wollina, 2015). ...
... 40 The alimentation of vegetarians is poor in carnitine, and for this reason a compensatory significant stimulation of hOCTN2 expression in oral mucosa has been observed. 41 Drugs such as β-lactam antibiotics, which possess a quaternary nitrogen as carnitine does, are able to interfere with carnitine transport by hOCTN2 and to cause carnitine deficiency. 42 Multidrug and toxic compound extrusion (MATE) proteins mediate the H + -coupled electroneutral exchange of endogenous and exogenous OCs as their final excretion step in the luminal membranes of the renal tubules and bile canaliculi. ...
The introduction of chemotherapy in the treatment of cancer is one of the most important achievements of modern medicine, even allowing the cure of some lethal diseases such as testicular cancer and other malignant neoplasms. The number and type of chemotherapeutic agents available have steadily increased and have developed until the introduction of targeted tumor therapy. It is now evident that transporters play an important role for determining toxicity of chemotherapeutic drugs not only against target but also against nontarget cells. This is of special importance for intracellularly active hydrophilic drugs, which cannot freely penetrate the plasma membrane. Because many important chemotherapeutic agents are substrates of transporters for organic cations, this review discusses the known interaction of these substances with these transporters. A particular focus is given to the role of transporters for organic cations in the development of side effects of chemotherapy with platinum derivatives and in the efficacy of recently developed tyrosine kinase inhibitors to specifically target cancer cells. It is evident that specific inhibition of uptake transporters may be a possible strategy to protect against undesired side effects of platinum derivatives without compromising their antitumor efficacy. These transporters are also important for efficient targeting of tyrosine kinase inhibitors to cancer cells. However, in order to achieve the aims of protecting from undesired toxicities and improving the specificity of uptake by tumor cells, an exact knowledge of transporter expression, function, regulation under normal and pathologic conditions, and of genetically and epigenetically regulation is mandatory.
Objectives: The aim of the present study was to compare the results of fractional microneedle radiofrequency (FMR) therapy in vegan and omnivorous participants. Methods: A total of 30 vegan and 30 omnivorous women who were treated with FMR therapy for combating aging were included in the study. The clinical results were examined in Months 3 and 6 based on the Fitzpatrick Wrinkle Scale (FWS). Individual satisfaction was investigated in Month 6 using the Patient's Global Impression of Change (PGIC). Results: At the onset of the treatment, there was no significant difference with regard to the FWS scores in both groups. The decrease in the FWS score was significantly lower in vegans after 3 months (p = 0.01). Vegans had worsened clinical outcomes by Month 6 (p = 0.01). The PGIC scores were significantly lower in vegans (p = 0.01). Conclusion: A vegan diet adversely affects the outcome of FMR therapy.
Background and objectives: Skin photoaging is related to extrinsic environmental exposures, mainly represented by ultraviolet radiation. One of the treatment options is laser resurfacing. As nutritional status is involved in cutaneous photodamage, we evaluated whether dietary patterns can also influence the response to facial resurfacing. Our prospective multicentric study involves three dermatologic centers specialized in laser therapy in northern Italy. The study aims to compare the outcome of a CO2 ablative laser therapy between omnivore and vegan patients. Study design/materials and methods: Fifty-three omnivore and fifty-three vegan women undergoing ultrapulsed CO2 resurfacing for photodamaged facial skin were enrolled in this study. Clinical improvement was evaluated 3 and 6 months after the treatment using the modified Dover score. Results: After laser treatment, vegans showed slower complete re-epithelialization (P < 0.001*) and disappearance of the erythema (P < 0.001*). After 3 and 6 months, vegans showed worse outcomes in terms of fine lines (P < 0.001* and P < 0.001*, respectively) and tactile roughness (P = 0.003* and P = 0.002*, respectively) compared with omnivores, while they did not differ in mottled pigmentation. Conclusions: The present study suggests that diet influences the clinical outcome of fractioned CO2 laser treatment. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.
Background: The aging of facial structures depends on genetic, anatomic, chronologic, and environmental factors that affect the skin and underlying tissues. Microfocused ultrasound with visualization (MFU-V) has emerged as a safe and effective treatment for skin laxity. As the nutritional status may contribute to skin aging, it would be interesting to evaluate whether different dietary patterns can also influence the response to MFU-V treatment for skin laxity. Aims: The aim of this study is to compare the outcome of MFU-V therapy between omnivore and vegan patients. Methods: Twenty-seven vegan and twenty-seven omnivorous women who underwent MFU-V treatment for laxity of lower face and neck were enrolled. The clinical outcome was evaluated using the FLR (Facial Laxity Rating) scale after 3 and 6 months from treatment. Results: At baseline, no significant differences were found in terms of FLR scale in both treated sites. After 3 months, reduction in FLR scale was significantly lower for vegans both on face (P = .04) and neck (P = .004). At 6 months, vegan patients had a worse clinical outcome on lower face (P = .001) and neck (P < .001). Conclusion: The present study suggests that a vegan diet may negatively influence the outcome of a MFU-V treatment.
Background: Postsurgical skin healing can result in different scars types, ranging from a fine line to pathologic scars, in relation to patients' intrinsic and extrinsic factors. Although the role of nutrition in influencing skin healing is known, no previous studies investigated if the vegan diet may affect postsurgical wounds. Objective: The aim of this study was to compare surgical scars between omnivore and vegan patients. Methods and materials: This is a prospective observational study. Twenty-one omnivore and 21 vegan patients who underwent surgical excision of a nonmelanoma skin cancer were enrolled. Postsurgical complications and scar quality were evaluated using the modified Scar Cosmesis Assessment and Rating (SCAR) scale. Results: Vegans showed a significantly lower mean serum iron level (p < .001) and vitamin B12 (p < .001). Wound diastasis was more frequent in vegans (p = .008). After 6 months, vegan patients had a higher modified SCAR score than omnivores (p < .001), showing the worst scar spread (p < .001), more frequent atrophic scars (p < .001), and worse overall impression (p < .001). Conclusion: This study suggests that a vegan diet may negatively influence the outcome of surgical scars.
Background: Photodynamic therapy (PDT) is an approved and effective treatment for actinic keratosis (AK). The time of complete skin healing is estimated to range between 5-10 days, but the role of nutrition in influencing it has never been evaluated. Objective: The aim of this study was to compare the time of skin healing and side effects in omnivores and vegans treated with PDT for AK. Materials and methods: Thirty omnivore and thirty vegan patients, treated with PDT for AK, were enrolled. Side effects, according to Local Skin Response (LSR) score, were compared after 3, 7 and 30 days; the time of complete skin healing was recorded. Results: At day 3, day 7 and day 30 post treatment vegan group showed higher total LSR score (p = 0,008, p < 0.001, p < 0.001 respectively), highlighting higher oedema and vesiculation at day 3 (p < 0.001, p = 0.002 respectively), erythema, desquamation, oedema and vesiculation at day 7 (p < 0.001, p < 0.001, p < 0.001, p < 0.001 respectively) and erythema and desquamation after 30 days (p < 0.001, p < 0.001 respectively). The difference of complete skin healing was statistically significant (p < 0.001). Conclusion: The present study suggests that diet may have a prognostic and predictive role on PDT outcomes in term of side effects and time of skin repair.
Full-text available
The effects of recombinant human tumor necrosis factor alpha (TNF alpha) on collagen production and gene expression in cultured fibroblasts were studied. Cells were labeled with [3H]proline, and the radioactivity of collagenase-sensitive and -resistant proteins were used to calculate the rates of protein production. The net production of collagen relative to total proteins was inhibited by TNF alpha (0-1.2 nM) in a dose- and time-related manner. The specific activities of the free [3H]proline pool, which were similar in control and TNF alpha-treated cells, were used to calculate the absolute rates of protein production. The absolute rate of collagen production was decreased by 50% in the presence of 1.2 nM TNF alpha during 24-h incubations (851 +/- 104 versus 426 +/- 39 pmol/micrograms of DNA/h; p less than 0.01), whereas noncollagen protein production and the rate of procollagen secretion were unchanged. We found no evidence of cellular toxicity in cultured cells treated with TNF alpha. In addition, TNF alpha did not affect cell proliferation as determined by [6-3H]thymidine incorporation into DNA. Most of the collagen produced by the cultured fibroblasts was type I. Using hybridization with specific DNA probes there was an approximately 50% decrease in the quantity of procollagen alpha 1(I) mRNA, without changes in the quantity of alpha tubulin mRNA or the size of the transcripts, in cells incubated with TNF alpha. Interleukin-1 (2.5 ng/ml) also decreased the levels of procollagen alpha 1(I) mRNA by approximately 50%. Cycloheximide (0.1 mM), an inhibitor of protein synthesis, blocked the inhibitory effect of both TNF alpha and interleukin-1 on procollagen alpha 1(I) mRNA. Nuclear run-off assays demonstrated that TNF alpha decreased procollagen alpha 1(I) transcriptional activity by 50% and had no effects on alpha tubulin gene transcription. Thus, TNF alpha decreases collagen gene transcription, collagen mRNA levels, and collagen production in cultured fibroblasts.
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The enzyme activity and the expression (protein and mRNA concentrations) of genes encoding for hepatic carnitine palmitoyl-transferases (CPT) I and II were studied during neonatal development, in response to nutritional state at weaning and during the fed-starved transition in adult rats. The activity, the protein concentration and the level of mRNA encoding CPT I are low in foetal-rat liver and increase 5-fold during the first day of extra-uterine life. The activity and gene expression of CPT I are high during the entire suckling period, in the liver of 30-day-old rats weaned at 20 days on to a high-fat diet and in the liver of 48 h-starved adult rats. The activity and CPT I gene expression are markedly decreased in the liver of rats weaned on to a high-carbohydrate diet. By contrast, the activity, the protein concentration and the level of mRNA encoding CPT II are already high in the liver of term foetuses and remain at this level throughout the suckling period, irrespective of the nutritional state of the animals either at weaning or in the adult.
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Interferon-alpha (IFN-alpha) induces cytogenetic responses of variable degree in patients with chronic myelogenous leukemia (CML). We sought to establish the relationship between BCR-ABL transcript numbers measured by competitive 2-step reverse transcription polymerase chain reaction (RT-PCR) and cytogenetic status in CML patients treated with IFN-alpha. A total of 250 peripheral blood and 55 bone marrow samples with 127 Philadelphia chromosome positive (Ph+) and 6 Ph-/BCR-ABL+ CML patients were investigated. Twenty-one patients were studied at diagnosis with IFN-alpha, 24 had a complete cytogenetic response, 21 a partial response, 12 a minor response, 26 no response, and 23 were unknown. Using nested RT-PCR, all 305 samples were positive for BCR-ABL transcripts. To standardize results for variability in RNA and cDNA quantity and quality, we quantified total ABL transcripts in each sample as internal control. The validity of ABL as internal control was shown by comparison with glucose-6-phosphate dehydrogenase transcript levels in 145 samples. The median BCR-ABL transcript numbers (and BCR-ABL/ABL ratios expressed as percentages) were 400/micrograms RNA (O.04%) in complete responders, 20,500/micrograms RNA (7.1%) in partial responders, 170,000/micrograms RNA (21.0%) in minor responders, and 430,000/micrograms RNA (58.7%) in nonresponders (P < .001). The cytogenetic results correlated with the BCR-ABL transcript numbers (r = .82; P < .001) and BCR-ABL/ABL ratios (r = .84; P < .001). Grouping the ratios BCR-ABL/ABL as less than 2%, 2% to 14% and greater than 14% to compare with cytogenetic complete response, partial response, and minor/nonresponse, the concordance between the two methods was 82% (chi2 P< .0001). We conclude that quantitative PCR with internal controls is as sensitive and reliable method for monitoring patients on IFN-alpha and reduces the need for repeated marrow investigations.
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To describe methods and dietary habits of a large population cohort. Prospective assessment of diet using diet diaries and food-frequency questionnaires, and biomarkers of diet in 24-h urine collections and blood samples. Free living individuals aged 45 to 75 years living in Norfolk, UK. Food and nutrient intake from a food-frequency questionnaire on 23 003 men and women, and from a 7-day diet diary from 2117 men and women. Nitrogen, sodium and potassium excretion was obtained from single 24-h urine samples from 300 individuals in the EPIC cohort. Plasma vitamin C was measured for 20 846 men and women. The food-frequency questionnaire (FFQ) and the food diary were able to determine differences in foods and nutrients between the sexes and were reliable as judged by repeated administrations of each method. Plasma vitamin C was significantly higher in women than men. There were significant differences in mean intake of all nutrients measured by the two different methods in women but less so in men. The questionnaire overestimated dairy products and vegetables in both men and women when compared with intakes derived from the diary, but underestimated cereal and meat intake in men. There were some consistent trends with age in food and nutrient intakes assessed by both methods, particularly in men. Correlation coefficients between dietary intake assessed from the diary and excretion of nitrogen and potassium in a single 24-h urine sample ranged from 0.36 to 0.47. Those comparing urine excretion and intake assessed from the FFQ were 0.09 to 0.26. The correlations between plasma vitamin C and dietary intake from the first FFQ, 24-h recall or diary were 0.28, 0.35 and 0.40. EPIC Norfolk is one of the largest epidemiological studies of nutrition in the UK and the largest on which plasma vitamin C has been obtained. Methods for obtaining food and nutrient intake are described in detail. The results shown here for food and nutrient intakes can be compared with results from other population studies utilising different methods of assessing dietary intake. The utility of different methods used in different settings within the main EPIC cohort is described. The FFQ is to be used particularly in pooled analyses of risk from diet in relation to cancer incidence within the larger European EPIC study, where measurement error is more likely to be overcome by large dietary heterogeneity on an international basis. Findings in the UK, where dietary variation between individuals is smaller and hence the need to use a more accurate individual method greater, will be derived from the 7-day diary information on a nested case-control basis. 24-h recalls can be used in the event that diary information should not be forthcoming from some eventual cases. Combinations of results utilising all dietary methods and biomarkers may also be possible.
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Previous studies suggest that the failing heart reactivates fetal genes and reverts to a fetal pattern of energy substrate metabolism. We tested this hypothesis by examining metabolic gene expression profiles in the fetal, nonfailing, and failing human heart. Human left ventricular tissue (apex) was obtained from 9 fetal, 10 nonfailing, and 10 failing adult hearts. Using quantitative reverse transcription-polymerase chain reaction, we measured transcript levels of atrial natriuretic factor, myosin heavy chain-alpha and -beta, and 13 key regulators of energy substrate metabolism, of which 3 are considered "adult" isoforms (GLUT4, mGS, mCPT-I) and 3 are considered "fetal" isoforms (GLUT1, lGS, and lCPT-I), primarily through previous studies in rodent models. Compared with the nonfailing adult heart, steady-state mRNA levels of atrial natriuretic factor were increased in both the fetal and the failing heart. The 2 myosin heavy chain isoforms showed the highest expression level in the nonfailing heart. Transcript levels of most of the metabolic genes were higher in the nonfailing heart than the fetal heart. Adult isogenes predominated in all groups and always showed a greater induction than the fetal isogenes in the nonfailing heart compared with the fetal heart. In the failing heart, the expression of metabolic genes decreased to the same levels as in the fetal heart. In the human heart, metabolic genes exist as constitutive and inducible forms. The failing adult heart reverts to a fetal metabolic gene profile by downregulating adult gene transcripts rather than by upregulating fetal genes.
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Aging affects oxidative metabolism in liver and other tissues. Carnitine acyltransferases are key enzymes of this process in mitochondria. As previously shown, the rate of transcription and activity of carnitine palmitoyltransferase CPT1 are also related to carnitine levels. In this study we compared the effect of dietary l-carnitine (100 mg l-carnitine/kg body weight/day over 3 months) on liver enzymes of aged rats (months 21-24) to adult animals (months 6-9) and age-related controls for both groups. The transcription rate of CPT1, CPT2, and carnitine acetyltransferase (CRAT) was determined by quantitative reverse transcription real-time PCR (RTQPCR) and compared to the activity of the CPT1A enzyme. The results showed that the transcription rates of CPT1, CPT2, and CRAT were similar in aged and adult control animals. Carnitine-fed old rats had a significant (p<0.05) 8-12-fold higher mean transcription rate of CPT1 and CRAT compared to aged controls, adult carnitine-fed animals, and adult controls, whereas the transcription rate of CPT2 was stimulated 2-3-fold in carnitine-fed animals of both age groups. With regard to the enzymatic activity of CPT1 there was a 1.5-fold increase in the old carnitine group compared to all other groups. RNA in situ hybridization also indicated an enhanced expression of CPT1A in hepatocytes from l-carnitine-supplemented animals. These results suggest that l-carnitine stimulates transcription of CPT1, CPT2, and CRAT as well as the enzyme activity of CPT1 in the livers of aged rats.
Conference Paper
Aging is associated with remarkable changes in body composition. Loss of skeletal muscle, a process called sarcopenia, is a prominent feature of these changes. In addition, gains in total body fat and visceral fat content continue into late life. The cause of sarcopenia is likely a result of a number of changes that also occur with aging. These include reduced levels of physical activity, changing endocrine function (reduced testosterone, growth hormone, and estrogen levels), insulin resistance, and increased dietary protein needs. Healthy free-living elderly men and women have been shown to accommodate to the Recommended Dietary Allowance (RDA) for protein of 0.8 g . kg(-1) . d(-1) with a continued decrease in urinary nitrogen excretion and reduced muscle mass. While many elderly people consume adequate amounts of protein, many older people have a reduced appetite and consume less than the protein RDA, likely resulting in an accelerated rate of sarcopenia. One important strategy that counters sarcopenia is strength conditioning. Strength conditioning will result in an increase in muscle size and this increase in size is largely the result of increased contractile proteins. The mechanisms by which the mechanical events stimulate an increase in RNA synthesis and subsequent protein synthesis are not well understood. Lifting weight requires that a muscle shorten as it produces force (concentric contraction). Lowering the weight, on the other hand, forces the muscle to lengthen as it produces force (eccentric contraction). These lengthening muscle contractions have been shown to produce ultrastructural damage (microscopic tears in contractile proteins muscle cells) that may stimulate increased muscle protein turnover. This muscle damage produces a cascade of metabolic events which is similar to an acute phase response and includes complement activation, mobilization of neutrophils, increased circulating an skeletal muscle interleukin-1, macrophage accumulation in muscle, and an increase in muscle protein synthesis and degradation. While endurance exercise increases the oxidation of essential amino acids and increases the requirement for dietary protein, resistance exercise results in a decrease in nitrogen excretion, lowering dietary protein needs. This increased efficiency of protein use may be important for wasting diseases such as HIV infection and cancer and particularly in elderly people suffering from sarcopenia. Research has indicated that increased dietary protein intake (up to 1.6 g protein . kg(-1) . d(-1)) may enhance the hypertrophic response to resistance exercise. It has also been demonstrated that in very old men and women the use of a protein-calorie supplement was associated with greater strength and muscle mass gains than did the use of placebo.
Background Cobalamin deficiency is prevalent in vegetarians and has been associated with increased risk of osteoporosis. Aim of the study To examine the association between cobalamin status and bone mineral density in adolescents formerly fed a macrobiotic diet and in their counterparts. Methods In this cross–sectional study bone mineral density (BMD) and bone mineral content (BMC) were determined by DEXA in 73 adolescents (9–15 y) who were fed a macrobiotic diet up to the age of 6 years followed by a lacto–(–ovo–) vegetarian or omnivorous diet. Data from 94 adolescents having consumed an omnivorous diet throughout their lives were used as controls. Serum concentrations of cobalamin, methylmalonic acid (MMA) and homocysteine were measured and calcium intake was assessed by questionnaire. Analysis of covariance (MANCOVA) was performed to calculate adjusted means for vitamin B12 and MMA for low and normal BMC and BMD groups. Results Serum cobalamin concentrations were significantly lower (geometric mean (GM) 246 pmol/L vs. 469 pmol/L) and MMA concentrations were significantly higher (GM 0.27 µmol/L vs. 0.16 µmol/L) in the formerly macrobiotic–fed adolescents compared to their counterparts. In the total study population, after adjusting for height, weight, bone area, percent lean body mass, age, puberty and calcium intake, serum MMA was significantly higher in subjects with a low BMD (p = 0.0003) than in subjects with a normal BMD. Vitamin B12 was significantly lower in the group with low BMD (p = 0.0035) or BMC (p = 0.0038) than in the group with normal BMD or BMC. When analyses were restricted to the group of formerly macrobiotic–fed adolescents, MMA concentration remained higher in the low BMD group compared to the normal BMD group. Conclusion In adolescents, signs of an impaired cobalamin status, as judged by elevated concentrations of methylmalonic acid, were associated with low BMD. This was especially true in adolescents fed a macrobiotic diet during the first years of life, where cobalamin deficiency was more prominent.
A 12-year old boy suffered episodes of vomiting, lethargy, and hypoglycaemia from the age of 1 year. Adhering to a vegetarian diet caused an increase in frequency and severity of the attacks. It was found that he was suffering from systemic carnitine deficiency that responded promptly to treatment with L-carnitine.
In this short review we summarize the effect of age on glucose homeostasis. The concept of decreased glucose tolerance with increasing age is introduced, followed by evidence for this phenomenon. Specifically we review the evidence for changes in fasting glucose as a function of age and the effect of age on HbA1c. The role of age on hepatic glucose production and glucose uptake is then discussed in detail and we review the evidence that supports the concept that with advancing age hepatic glucose sensitivity to insulin is unaltered. We then review the large evidence for the role of age on the purported decrease in peripheral tissue sensitivity to insulin and conclude that the issue is unsettled. The decrease attributed to age is no longer significant when confounders are controlled for, the largest being obesity. We next present evidence that beta-cell sensitivity to glucose remains intact with aging. A review of age-related disorders due to hyperglycemia and confounding effects on the relationships of age and glucose tolerance is presented next. Finally we present new evidence that when the revised criteria for the diagnosis of type 2 diabetics as proposed by the American Diabetes Association and WHO are used, a greater percentage of the elderly will not be diagnosed. We conclude that, although glucose intolerance increases with aging, which is accompanied with other disorders, it is possible to ameliorate this effect with alteration of diet and exercise.