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Objectives International cutoff points for the diagnosis of sarcopenia are not applicable to the Chilean population due to previous evidence of a lower lean mass and strength in this population. Dual-energy x-ray absorptiometry is used to establish fat-free mass cutoff points to define sarcopenia in the Chilean population and analyze its association with handgrip strength in older adults. Methods Appendicular fat-free mass (AFFM) was calculated from 4062 dual-energy x-ray absorptiometries of healthy Chileans, ages 18 to 99 y. Possible cutoff points for sarcopenia were obtained using four methods: A) Normative, –2 standard deviation (SD) below mean AFFM/height2 (AFFMI) of adults age Results Using the normative method, sarcopenia was defined as an AFFMI Conclusions The AFFM of young Chileans is lower than that reported in Western countries but similar to Latin American data; therefore, the use of the traditional normative method would not be appropriate with –2 SD to establish cutoff points, and using –1 DS resulted in values that are higher than Baumgartner's. Stratification is advantageous because this method throws expected values of AFFM for each population; however, overdiagnosis of sarcopenia is a possibility and thus the method requires a representative sample. The percentage method is simple and showed the expected decrease of muscle mass with age, and also correlated well with handgrip strength in elderly women. Thus, this method represented our method of choice to detect sarcopenia.

Changes in DNA methylation, one of the most studied epigenetic mechanisms, are considered an initial marker for early cancer detection. We evaluated how availability of dietary factors (folates and vitamin B12) involved in one-carbon metabolism may contribute to DNA methylation changes of cancer-related genes in human subjects. We studied, by pyrosequencing, the methylation of tumor suppressor gene p16, DNA repair genes MLH1 and MGMT, and the repetitive element LINE-1 (as a surrogate for global DNA methylation), in blood of elderly individuals (n = 249) who had been exposed to folic acid (FA) through FA-fortified wheat flour during the last 12 years. We found that serum folate and to a lesser extent, vitamin B12 concentrations, were significantly correlated with DNA methylation of p16, MLH1, and MGMT, but not with LINE-1. High serum folate concentrations (>45.3 nmol/L) were present in 31.1% of the participants. Although the methylated fraction of CpG sites in p16, MLH1, and MGMT was low (1.17–3.8%), high folate concentrations were significantly associated with methylation at the 3rd tertile of specific CpG sites in all genes with OR between 1.97 and 4.17. This study shows that a public policy, like food fortification with FA that increases circulating serum folate levels, could affect methylation levels of specific genes linked to cancer risk. Our present results deserve additional studies to clarify the real impact of high FA levels for risk of cancer in a whole population chronically exposed to a fortified food such as wheat flour. ISRCTN 48153354 and ISRCTN 02694183

Summary Problem Up to 35% of hospitalized patients may experience functional decline during or after hospitalization. Subjective Global Assessment (SGA) and handgrip strength at admission, have been proposed as simple and accessible tools to predict functional decline, but there are few studies in hospitalized patients to confirm these findings. Objective To assess the predictive value of handgrip strength at hospital admission, on functional decline after 30 days. Methods 125 non-critical patients hospitalized for medical and surgical conditions, were studied in El Pino hospital in Santiago, Chile. Upon admission, nutritional status was assessed by SGA, functional status through the Karnofsky index (KI), and handgrip strength by dynamometry. Change in functionality was assessed by the difference between KI at admission and 30 days later. Multivariate logistic regression models were used to establish associations between the variables at hospital admission, and subsequent functional decline. Results Thirty days post-hospital admission, 28.8% of the sample showed functional decline. In a multivariate analysis, only handgrip strength was associated with this decline (β = −0.025, OR = 0.974 (CI 0.956–0.992), p = 0.007). Conclusions Handgrip strength upon hospital admission can be a useful independent and early method to predict deterioration of functional status during hospitalization.

Objectives Predictive equations are frequently used to estimate resting energy expenditure (REE) because indirect calorimetry (IC) is not always available and is expensive. The aim of this study was to determine the concordance between the estimation of REE using predictive equations and its measurement by IC. Methods This was an analysis of the registry of indirect calorimetry performed in non-hospitalized participants. Harris–Benedict, FAO/WHO/UNU, Mifflin St. Jeor, and European Society for Clinical Nutrition and Metabolism (ESPEN) equations were used to estimate REE in these individuals. The concordance between measured and estimated REE using real, ideal, and adjusted weight was calculated using the concordance coefficient analysis of Lin and Bland– Altman plots in all participants and in subgroups separated according to their body mass index. Results We retrieved 680 measurements and discarded 247 that did not comply with the inclusion criteria. Thus, we studied 433 participants ages 36 y (29–48 y). Of the participants, 341 were women (79%) and the participants had a body mass index (BMI) of 30 kg/m2 (26.7–33.1 kg/m2). All predictive equations had concordance values 10% ranged from 36% to 87%. The ESPEN equation had the greater proportion of erroneous estimations of REE in all participants and BMI subgroups when real weight was used. Conclusions We observed a low level of concordance between REE estimated using predictive equations and measured by IC. These results should alert clinicians about the inaccuracy of predictive equations.

Objective Omega-3 fatty acids may reduce albuminuria and cardiovascular risk factors in patients with chronic kidney disease (CKD). We aimed to assess the effects of omega-3 fatty acid supplementation on albuminuria, blood pressure, pulse wave velocity, and inflammatory markers in patients with CKD. Methods Patients with CKD and a urine albumin excretion of at least 30 mg/g creatinine were supplemented for 3 months with 3,666 mg/day of docosahexaenoic and eicosapentaenoic acids or a corn oil supplement. The study was double blind. At baseline, 6 weeks, and 12 weeks, fasting blood and morning spot urine samples were obtained. Blood pressure, carotid intima media thickness, and pulse wave velocity were measured. The main outcome measure was a reduction of ≥20% in urine albumin. Results One hundred patients were randomized (50 received omega-3 fatty acids and 50 received corn oil). Four patients who received omega-3 fatty acids and 5 who received vegetable oil were lost to follow-up. In patients receiving omega-3 fatty acids, the omega-3 index increased from 3.08 (2.32-3.81) to 5.48 (3.045-7.04) percent. A 20% reduction in urine albumin excretion was observed in 13 participants of the control group and 19 participants of omega-3 group (Fisher's exact P = .274). However, the supplement had a significant and positive effect on pulse wave velocity and triglyceride level. Conclusion An omega-3 fatty acid supplement of 3,666 mg/day did not modify urine albumin excretion in patients with CKD but did improve pulse wave velocity and serum triglyceride levels.

espanolIntroduccion: la restriccion proteica es fundamental en el manejo de la enfermedad renal cronica. Objetivo: evaluamos la utilidad de la medicion de nitrogeno ureico urinario como marcador de restriccion proteica. Metodos: participantes jovenes sanos fueron divididos aleatoriamente en dos grupos. Un grupo recibio una dieta con 30 kcal/dia/kg peso corporal y 1 g/proteina/dia/kg peso corporal y el otro recibio una dieta con la misma cantidad de calorias pero con 0,6 g/kg peso corporal de proteina. Al inicio, a los siete y a los 14 dias, los participantes respondieron una encuesta dietaria de recordatorio de 24 horas. Ademas, recolectaron orina de 24 horas y se les tomo una muestra aislada de orina al comienzo y a los 14 dias de la intervencion para medir creatinina y nitrogeno ureico. Resultados: cuarenta y un participantes de 29 ± 5 anos completaron el estudio. El grupo que consumio 0,6 g/kg de proteinas redujo su ingesta proteica de 0,88 ± 0,06 a 0,59 ± 0,05 g/kg/dia durante la intervencion. En este grupo se observo una reduccion significativa en la excrecion urinaria de nitrogeno ureico en 24 horas. No se observo tal reduccion en el grupo que consumio 1 g/kg de proteinas. La tasa de probabilidad de detectar una reduccion en la ingesta proteica en las encuestas dietarias, cuando se observaba una disminucion en la excrecion urinaria de nitrogeno ureico/mg creatinina de 24 horas, fue de 5,75 (intervalos de confianza de 95% = 1,29-25,55, p = 0,02). No hubo cambios significativos en la excrecion de nitrogeno ureico en las muestras aisladas de orina. Conclusion: las mediciones repetidas de nitrogeno ureico urinario en 24 horas son un marcador de restriccion dietaria de proteinas. EnglishBackground: protein restriction is the mainstay of dietary management of chronic kidney disease. Aim: to assess the usefulness of urine urea nitrogen measurement as a marker of protein restriction. Methods: healthy young participants were randomly divided in two groups. During 14 days, one group received a diet containing 30 kcal/kg body weight and 1 g protein/kg body weight and the other group received a diet with the same amount of calories and 0.6 g/kg of proteins. At baseline, seven days and 14 days, 24 h dietary recalls were answered by the participants. They collected 24 hour urine and provided spot urine samples at baseline and at the end of the intervention, to measure creatinine and urea nitrogen. Results: forty-one participants aged 29 ± 5 years completed the follow-up. According to 24h dietary recalls, the group receiving 0.6 g/kg protein reduced significantly the protein intake during the intervention from 0.88 ± 0.06 to 0.59 ± 0.05 g/kg/day. A significant reduction in 24 h urea nitrogen excretion was also observed in this group. In the group receiving 1 g/kg of protein, no significant changes in 24 h urea nitrogen excretion were observed. Among all participants, the odds ratio of observing a reduction in protein intake in the dietary survey was 5.75 (95% confidence intervals 1.29-25.55, p = 0.02), when a reduction in 24 h urea nitrogen excretion corrected by creatinine was observed. No changes were observed in urea nitrogen excretion in spot urine samples. Conclusions: repeated urea nitrogen excretion measured in 24 h urine samples can be a reliable indicator of dietary protein restriction.

Introduction Muscle mass and function are among the most relevant factors that contribute to an optimal quality of life, and are strong predictors of mortality in the elderly. Loss of lean tissues and deterioration of muscle function have been described as one of the many complications of type 2 diabetes mellitus (DM2), but most studies do not isolate age as an intervening factor. Aim To study whether adult DM2 patients up to 60 years of age have decreased muscle mass and function compared with healthy non-diabetic (ND) subjects of similar age. Methodology Appendicular fat-free mass (ApFFM) by dual X-ray absorptiometry (DEXA), handgrip strength (HS), quadriceps strength (QS), 12 min walking capacity (12MW) and the Timed Up and Go test (TUG) were measured in 100 DM2 patients and 39 ND controls. Muscle quality, or the ratio between lean mass and muscle strength of upper and lower limbs, and the functional limitations associated with pain and stiffness assessed according to the Western Ontario and McMaster Universities Arthrosis Index (WOMAC) were also recorded. Specific tests were performed to rule out microvascular diabetic complications (retinal and peripheral nerves), metabolic control, kidney function and vitamin D status and examine their association with ApFFM and function. Results ApFFM was significantly higher among DM2 female patients and lower among diabetic men. However opposite results were obtained when individual values were corrected for body mass index (BMI), specifically among women, who were more likely to be obese. As for muscle strength and global functionality tests, significantly better performances in TUG, 12MW, QS and HS were observed among ND subjects of both sexes. These differences prevailed even after excluding diabetic patients with microvascular complications as well as those with more than 10 years of diabetes. Muscle quality was also significantly better among ND women. Higher scores of pain and stiffness in the WOMAC scale correlated with 12MW and TUG in both groups but did not correlate with ApFFM. Conclusions We found a clear deterioration of lean mass and muscle functions among adult DM2 patients of up to 60 years old, independent of length of disease, metabolic control, vitamin D status and presence of microvascular complications and pain.

Background Sedentariness may be an important risk factor for sarcopenia. The aim of this work was to assess the association between muscle mass and strength and markers of usual physical activity such as activity energy expenditure and peak oxygen uptake. Methods Young and old participants were assessed measuring body composition by DEXA (double beam X ray absorptiometry), handgrip strength, peak oxygen consumption and workload during an exercise calorimetry in a braked cycle ergometer and a 72 h activity energy expenditure using Actiheart actigraphs. A heart rate/energy expenditure curve derived from the exercise calorimetry was used to calibrate each actigraph. Sarcopenia was defined as having an appendicular fat free mass index below 7.5 kg/m2 and 5.6 kg/m2 in men and women respectively, or a handgrip strength z score below 1, using local normal data or having both parameters below the cutoff points. Results We analyzed data from 192 assessments performed in participants aged 22 to 88 years (106 women). Sarcopenic participants (as determined by muscle mass, strength or both) had a significantly lower peak oxygen uptake and work load and a significantly lower activity energy expenditure. When analyzing lean mass and strength as continuous variables, peak oxygen consumption was a significant predictor of fat free mass in men. Among women, the association was observed only when percentage of muscle mass was expressed as a z score. Conclusions Activity energy expenditure and peak oxygen consumption are associated with a lower muscle mass and the presence of sarcopenia and should be considered as risk factors for this condition.

Background: the prognostic value of maximal inspiratory and expiratory pressures on functional capacity and mortality of hospitalized patients are not well established. Aim: to evaluate the prognostic value of respiratory pressures in hospitalized patients. Methods: patients admitted to a general hospital in Santiago-Chile were prospectively studied. Within 48 hours of admission, handgrip strength and inspiratory and expiratory pressures were measured. Subjective global assessment of nutritional status (SGA) was determined and Apache II score was calculated. Functional status was assessed using the Karnofski index. Patients were followed for a period of 30 days. Mortality and decline in functional capacity, defined as a reduction in at least two stages of the Karnofski index were determined. Normal values for handgrip strength and respiratory pressures were obtained in 366 healthy subjects aged 20 to 89 years, thus the results obtained in patients were expressed as age and sex matched z-scores. Results: one hundred and eight patients were recruited and 18 had to be excluded. Thus, 90 patients aged 58 ± 16 years (46 females) were studied. During the observation period, six patients died and nine experienced a decline in functional status. Patients who died had significantly lower maximal inspiratory and expiratory pressures, hand grip strength and worse SGA. Logistic regression analysis only accepted maximal expiratory pressure expressed as z-score as a predictor of mortality. In addition, it was the only significant predictor of death or functional decline. Conclusions: maximal expiratory pressure on admission was a predictor of death or functional decline at 30 days.

OBJECTIVETo determine whether probiotic prophylaxes reduce the odds ofClostridium difficileinfection (CDI) in adults and children.DESIGNIndividual participant data (IPD) meta-analysis of randomized controlled trials (RCTs), adjusting for risk factors.METHODSWe searched 6 databases and 11 grey literature sources from inception to April 2016. We identified 32 RCTs (n=8,713); among them, 18 RCTs provided IPD (n=6,851 participants) comparing probiotic prophylaxis to placebo or no treatment (standard care). One reviewer prepared the IPD, and 2 reviewers extracted data, rated study quality, and graded evidence quality.RESULTSProbiotics reduced CDI odds in the unadjusted model (n=6,645; odds ratio [OR] 0.37; 95% confidence interval [CI], 0.25–0.55) and the adjusted model (n=5,074; OR, 0.35; 95% CI, 0.23–0.55). Using 2 or more antibiotics increased the odds of CDI (OR, 2.20; 95% CI, 1.11–4.37), whereas age, sex, hospitalization status, and high-risk antibiotic exposure did not. Adjusted subgroup analyses suggested that, compared to no probiotics, multispecies probiotics were more beneficial than single-species probiotics, as was using probiotics in clinical settings where the CDI risk is ≥5%. Of 18 studies, 14 reported adverse events. In 11 of these 14 studies, the adverse events were retained in the adjusted model. Odds for serious adverse events were similar for both groups in the unadjusted analyses (n=4,990; OR, 1.06; 95% CI, 0.89–1.26) and adjusted analyses (n=4,718; OR, 1.06; 95% CI, 0.89–1.28). Missing outcome data for CDI ranged from 0% to 25.8%. Our analyses were robust to a sensitivity analysis for missingness.CONCLUSIONSModerate quality (ie, certainty) evidence suggests that probiotic prophylaxis may be a useful and safe CDI prevention strategy, particularly among participants taking 2 or more antibiotics and in hospital settings where the risk of CDI is ≥5%.TRIAL REGISTRATIONPROSPERO 2015 identifier: CRD42015015701Infect Control Hosp Epidemiol2018;771–781

Chronic renal disease (CRD) in its pre-dialysis stage is an important risk factor for mortality among adults. The aim of this study was to assess the effects of CRD on mortality among consultants in Chilean public primary care clinics. We obtained information about serum creatinine, urinary albumin excretion (UAE), blood pressure, and body mass index of 5224 consultants [3379 females aged 67 (59-75) years and 1845 males aged 68 (59-75) years] in three clinics of Metropolitan Santiago. Kaplan-Meier curves and Cox proportional hazard regression models were used to determine risk factors for mortality, determined 41 months after obtaining the blood samples. During the follow-up period, 262 patients died (33% due to circulatory causes and 29% due to tumors). Kaplan-Meier curves showed that there was a significant association between survival, estimated glomerular filtration rate, and UAE. Cox models showed that serum creatinine, UAE, a lower body mass index, and a history of diabetes were significant mortality predictors. A sensitivity analysis performed eliminating extreme ages (less than 50 and more than 80 years), included high diastolic pressure as a predictor of survival. We conclude that among patients with CRD in its pre-dialysis stage, UAE is an important predictor of survival, along with serum creatinine. A low body mass index was associated with a higher mortality.

Immunosenescence has been described as age-associated changes in the immune function which are thought to be responsible for the increased morbidity with age. Human Natural Killer (NK) cells are a specialized heterogeneous subpopulation of lymphocytes involved in immune defense against tumor and microbial diseases. Interestingly, aging-related NK cell dysfunction is associated with features of aging such as tumor incidence, reduced vaccination efficacy, and short survival due to infection. It is known that NK cell effector functions are critically dependent on cytokines and metabolic activity. Our aim was to determine whether there is a difference in purified human NK cell function in response to high concentration of IL-2 between young and elder donors. Here, we report that the stimulation of human NK cells with IL-2 (2000 U/mL) enhance NK cell cytotoxic activity from both young and elderly donors. However, while NK cells from young people responded to IL-2 signaling by increasing mitochondrial mass and mitochondrial membrane potential, no increase in these mitochondrial functional parameters was seen in purified NK cells from elderly subjects. Moreover, as purified NK cells from the young exhibited an almost three-fold increase in PGC-1α expression after IL-2 (2000 U/mL) stimulation, PGC-1α expression was inhibited in purified NK cells from elders. Furthermore, this response upon PGC-1α expression after IL-2 stimulation promoted an increase in ROS production in NK cells from elderly humans, while no increase in ROS production was observed in NK cells of young donors. Our data show that IL-2 stimulates NK cell effector function through a signaling pathway which involves a PGC-1α-dependent mitochondrial function in young NK cells, however it seems that NK cells from older donors exhibit an altered IL-2 signaling which affects mitochondrial function associated with an increased production of ROS which could represent a feature of NK cell senescence.

Background Lack of weight gain throughout adult life could mimic the beneficial effects of energy restriction in humans. The present study aimed to assess the effects of weight stability or gain, over a period of 10 years, on telomere length, sirtuin 1 and 6 expression, and carotid intima media thickness. Methods We studied 148 healthy adults (age range 20–59 years; 101 females) who had an objective record of their weight 10 years before. They were classified as weight losers, weight maintainers, weight gainers and extreme weight gainers. A fasting blood sample was obtained for routine laboratory and isolation of peripheral blood mononuclear cells, to extract DNA and RNA, and to measure telomere length and sirtuin 1 and 6 expression, respectively. Carotid intima media thickness was measured by ultrasound. Body composition was measured by Dual-energy X-ray absorptiometry. Results In the 10-year period, 24 participants lost weight (17 females), 65 maintained weight (41 females), 25 gained weight (15 females) and 34 were extreme weight gainers (28 females). Female weight gainers had a higher body mass index, waist circumference, total body fat and homeostatic model assessment insulin resistance. Male weight gainers had a higher hip circumference and total body fat. No differences in telomere length, sirtuin 1 expression and carotid intima media thickness were observed between weight gainers and maintainers. Conclusions No effect of weight maintenance or gain was observed on metabolic and vascular markers of ageing.

Several cardiovascular diseases (CVD) observed in adulthood have been associated with environmental influences during fetal growth. Here, we show that maternal exposure to cadmium, a ubiquitously distributed heavy metal and main component of cigarette smoke is able to induce cardiovascular morpho-functional changes in the offspring at adult age. Heart morphology and vascular reactivity were evaluated in the adult offspring of rats exposed to 30 ppm of cadmium during pregnancy. Echocardiographic examination shows altered heart morphology characterized by a concentric left ventricular hypertrophy. Also, we observed a reduced endothelium-dependent reactivity in isolated aortic rings of adult offspring, while endothelium-independent reactivity remained unaltered. These effects were associated with an increase of hem-oxygenase 1 (HO-1) expression in the aortas of adult offspring. The expression of HO-1 was higher in females than males, a finding likely related to the sex-dependent expression of the vascular cell adhesion molecule 1 (VCAM-1), which was lower in the adult female. All these long-term consequences were observed along with normal birth weights and absence of detectable levels of cadmium in fetal and adult tissues of the offspring. In placental tissues however, cadmium levels were detected and correlated with increased NF-κB expression – a transcription factor sensitive to inflammation and oxidative stress – suggesting a placentary mechanism that affect genes related to the development of the cardiovascular system. Our results provide, for the first time, direct experimental evidence supporting that exposure to cadmium during pregnancy reprograms cardiovascular development of the offspring which in turn may conduce to a long term increased risk of CVD.

Introduction. The inhibition of angiotensin-converting enzyme could be useful to avoid sarcopenia in the elderly. Materials and methods. We compared in a prospective double blind trial, the effects of treatment with enalapril or nifedipine on muscle performance in hypertensive elderly subjects. Patients were followed for nine months, and at baseline, 4.5 months and the end of follow-up, quadriceps and hand grip muscle strength, walking capacity, timed up and go and the short physical performance test were measured. Results. During follow-up, more subjects on nifedipine than on enalapril discontinued the medication due to side-effects. No differences in the evolution of muscle strength, walking capacity or functional measures were observed. At nine months, plasma angiotensin-converting enzyme activity decreased by 6.0±2.5 U/L among patients on enalapril and increased by 8.5±4.2 U/L (p Conclusion. In this group of elderly subjects, enalapril was not superior to nifedipine with regard to the age-related decline of muscle performance.

Tissue accumulation of advanced glycation end products (AGEs) is associated with ageing, both in diabetics and nondiabetic subjects.The purpose of this study was to assess immunostaining for AGEs, specifically carboxymethyl-lysine (CML) and receptor for AGEs (RAGE), in muscle tissue of healthy male subjects differing in age and weight stability.Muscle tissue was obtained during hernia surgery in middle-aged men reporting weight maintenance (WM, n = 10) or weight gain (WG, n = 7), and also in 4 elderly men. Tissue inmunostaining for CML and RAGE was performed.Intensity of CML and RAGE staining were highly correlated (r = 0.84) and also significantly associated with weight change and age. Muscle AGEs accretion was statistically associated with muscle expression of oxidative injury (8-hydroxy-deoxyguanosine and 4-hydroxy-2-nonenal) and inflammatory markers (tumor necrosis factor-alpha).The increase of skeletal muscle AGEs/RAGE and markers of inflammation and oxidative injury in association with weight gain and old age suggest a pathogenic role of AGEs in weight gain and in sarcopenia of aging.

to asses Total Energy Expenditure (TEE) in healthy Chilean institutionalized or independently older people Methods: twenty seven young (27-30 years), 27 institutionalized (65 years old) and 27 free-living older (65 years old) participants were studied. Body composition was estimated by dual energy X-ray absorptiometry. Physical activity energy expenditure (AEE) and TEE were assessed using Actiheart accelerometers. The Mini Nutritional Assessment (MNA) was applied and Timed Up and Go (TUG) was measured.AEE was 171, 320 and 497 kcal/day in institutionalized, free living older and young participants, respectively (p0.01 between young and older participants). Both absolute TEE and TEE/RMR was higher in young people. Multiple regression analysis accepted age, MNA and TUG as significant predictors of AEE (r2 = 0.24 p0.01).AEE and PAL were lower among older people, with no differences by institutionalization.Objetivo: evaluar el Gasto Energético Total (GET) en ancianos sanos que viven institucionalizados o independientes en Chile. Método: se evaluaron veintisiete jovenes (27-30 años), 27 adultos mayores institucionalizados (65 años ) y 27 ancianos independientes (65 años). Se midió la composición corporal utilizando absorciometría bifotónica de rayos X. Se calculó el gasto energético por actividad física (GEAF) y el gasto energético total (GET) utilizando acelerómetros Actiheart; se aplicó Mini Nutritional Assessment (MNA) y se midió el Timed Up and Go (TUG). Resultados: el GEAF fue 171, 320 y 497 kcal/día en ancianos institucionalizados, independientes y jóvenes, respectivamente (p.

Adequate folate levels are essential for successful pregnancy outcomes. We aimed to study the relationship between placental mRNA and protein levels of folate transporters to birth weight.Placental folate transporters (FOLR1, RFC1 and HCP1/PCFT) mRNA and protein levels in basal (BP) and chorionic plate (CP) of small (SGA), appropriate (AGA) and large (LGA) for gestational age term infants (≥37 weeks gestation, n = 111) were determined by real-time PCR and Western blot respectively.FOLR1 and HCP1/PCFT mRNA were lower in both plates of SGA and LGA placentas compared to AGA (p 0.01) and RFC1 mRNA was lower only in CP (p 0.02). RFC1 protein levels were lower in BP of SGA (p 0.05) and LGA (p 0.01), and FOLR1 protein levels were lower in CP of SGA (p 0.02) and LGA (p 0.01) groups compared to AGA. HCP1/PCFT protein levels remained unchanged in all groups.Placentas of SGA and LGA groups showed a reduced mRNA expression and protein levels of folate transporters, with some differences depending on the location within the placenta (BP or CP). This suggests the presence of specific placental regulation mechanisms in gene expression that may be associated to birth weight.

Obesity is a risk factor for diabetes and its consequences, including accelerated ageing and mortality. The underlying factor could be accumulation of certain lipid moieties, such as ceramides (CER) and diacylgycerol (DAG) within muscle tissue, which are known to promote insulin resistance (IR), induce inflammation and oxidative injury, ultimately altering muscle function.First, to study the relationship between body composition and age (independent variables) with skeletal muscle accumulation of lipid species, oxidative injury and strength. Second, to analyze the relationship between muscle tissue metabolites and insulin resistance, inflammation and lymphocyte telomere length, the latter as an indicator of ageing.The sample included 56 healthy sedentary males, scheduled for inguinal hernia surgery, aged 27 to 80 y. Each individual was subject to anthropometric measurements, body composition assessment through radiologic densitometry (DEXA), measurement of handgrip and quadriceps strength, serum biochemical parameters (lipoproteins, creatinine, high sensitivity C reactive protein [hsCRP], fasting and post glucose insulin and glucose concentrations for calculation of IR through the Matsuda and HOMA-IR indexes), and extraction of peripheral leukocytes for measurement of telomere length. During the surgical procedure, a sample of muscle tissue was obtained (anterior abdominal oblique) in order to measure CER and DAG (and sub species according to chain length and saturation) by mass spectrometry, 4 hydroxy-2-nonenal adducts (4-HNE) using electron microscopy immunohistochemistry, and carboxymethyl-lisine (CML) by immunohistochemistry, the latter as indicators of oxidative stress (OS).Body mass index (BMI) of twenty six individuals was25 k/m2, while BMI of 7 was30 k/m2. Overweight/obese individuals, did not exhibit differences in skeletal muscle lipid metabolites, however total CER and specific long chain CER sub-species (20 and 22 carbon) increased significantly among individuals with a central fat distribution (n = 14) as well as in glucose intolerant subjects (n =23). A negative association was found between mononuclear leukocyte telomere length and 20 and 22 carbon CER (rho = - 0.4 and -0.5 0 p0.05). Muscle strength was not associated with any of the measured muscle metabolites or markers of OS. A multiple regression analysis accepted central abdominal fat and telomere length as significant predictors of CER (R2 = 0.28).An association was found between accumulation of specific ceramide species in muscle tissue and abdominal obesity, glucose intolerance and shortening of leukocyte telomeres, although not with muscle oxidative injury or dysfunction.

Summary Background and aims Weight maintenance within normal standards is recommended for prevention of conditions associated with oxidative injury. To compare oxidative damage in a post mitotic tissue, between adults differing in long-term energy balance. Methods During hernia surgery, a sample of skeletal muscle was obtained in 17 non-obese adults. Subjects were divided into two groups according to their self-reported weight change: weight maintainers (WM) reported 5 kg increment. Muscle immunohistochemistry for 8-hydroxy-deoxyguanosine (8OHdG), 4-Hydroxy-2-nonenal (4HNE), and TNF-α, as markers of oxidative injury and inflammation, were performed. As known positive controls for oxidative injury, we included 10 elderly subjects (66–101 yr). Anthropometric measures and blood samples for clinical laboratory and serum cytokines (TNF-α and IL-6) were obtained. Results 8OHdG was higher in WG compared with WM (149.1±16.2 versus 117.8±29.5, P = 0.03 ), and was associated with anthropometric indicators of fat accumulation. 4HNE was similar in WG compared with WM (10.9±7.6 versus 9.8±6.3) but noticeably higher in elderly subjects (21.5±15.3, P = 0.059 ). TNF-α protein in WG was higher compared with WM (114.0±41.7 versus 70.1±23.3, P = 0.025 ), and was associated with weight increase. Conclusions Moderate self-reported weight increase, and body fat accumulation, suggesting long-term positive energy balance is associated with muscle DNA oxidative injury and inflammation.

The aim was to assess the effects of resistance training and vitamin D supplementation on physical performance of healthy elderly subjects. Ninety-six subjects, aged 70 years or more with 25 OH vitamin D levels of 16 ng/ml or less, were randomized to a resistance training or control group. Trained and control groups were further randomized to receive in a double blind fashion, vitamin D 400 IU plus 800 mg of calcium per day or calcium alone. Subjects were followed for nine months. Serum 25 OH vitamin D increased from 12.4 ± 2.2 to 25.8 ± 6.5 ng/ml among subjects supplemented with vitamin D. Trained subjects had significant improvements in quadriceps muscle strength, the short physical performance test and timed up and go. The latter improved more in trained subjects supplemented with vitamin D. At the end of the follow up, gait speed was higher among subjects supplemented with vitamin (whether trained or not) than in non-supplemented subjects (838 ± 147 and 768 ± 127 m/12 min, respectively, p = 0.02). Romberg ratio was lower among supplemented controls than non-supplemented trained subjects (128 ± 40% and 144 ± 37%, respectively, p = 0.05). In conclusion, vitamin D supplementation improved gait speed and body sway, and training improved muscle strength.

Vitamin B-12 deficiency is prevalent among the elderly population but it is often unrecognized because the clinical manifestations are not present. Objective: To evaluate the effects of a nutritional supplement fortified with vitamin B-12 on well nourished, free-living elderly subjects. Patients and methods: Healthy elderly subjects attending two of four clinics were allocated to receive, over six months’ duration, a nutritional supplement with 3.8 µg of vitamin B-12. Subjects attending the other two clinics served as controls. Serum vitamin B-12 levels were measured at baseline and 6 months after the supplementation was started. Results: After 6 months of consuming the supplement fortified with vitamin B-12, serum B-12 concentration increased from 350.1 ± 166.5 pmol/L to 409.0 ± 166.1 and decreased in the control group from 319.4 ± 129.1 to 290.1 ± 135.7 (ANOVA, p < 0.005). Conclusion: A supplementation with 3.8 µg /day of vitamin B-12 led to significant improvements in the serum concentrations of vitamin B-12 in older persons.

Prebiotics increase intestinal levels of health-promoting bacteria implicated in decreasing pathogen colonization, stimulating immune functions and stabilizing gut barrier functions, parameters which are altered in burn patients. We propose that regular intake of a prebiotic, oligofructose (OF), might help to improve the altered gastrointestinal (GI) permeability observed in burn patients. A randomized, double-blind, controlled clinical trial was carried out in 41 burn patients (mean burn surface area=17.1+/-8.2%) who ingested daily 6 g of oligofructose (OF group) or sucrose as placebo (Control group) during 15 days. Gastrointestinal permeability to sucrose and lactulose/mannitol (L/M) was evaluated on days 1 (before treatment) 3, 7, 14 and 21. A permeability test was also performed in 18 healthy subjects as controls. Thirty-one patients completed the protocol (dropout rate=24.4%). Healthy subjects had a basal sucrose excretion of 21.3 mg (14.0-32.5 mg) and a basal L/M ratio of 0.017% (0.009-0.022%). Sucrose excretion increased 5-fold and L/M ratio 4.4-fold in burn patients on day 1 and these high levels of marker excretion decreased significantly throughout the study (p=0.016 and 0.000001, respectively). No differences between the OF and Control groups were observed for sucrose excretion or L/M ratio. In conclusion, the normalization of gastrointestinal permeability is not accelerated by prebiotic intake.

Immune response is impaired in the elderly. Our aim was to study the effects of a special nutritional formula on the immune response and response to influenza and pneumococcal vaccination in elderly subjects.Sixty healthy subjects agedor = 70 years, with a Mini Mental scoreor = 22 were studied. Half of the subjects received a special nutritional formula (in addition to the regular diet) providing, among other nutrients, 480 kcal, 31 g proteins, 120 IU vitamin E, 3.8 microg vitamin B12, 400 microg folic acid, 10(9) cfu Lactobacillus paracasei (NCC 2461), and 6 g of fructo-oligosaccharides. At 4 months of follow-up, subjects were vaccinated against influenza and pneumococcus. Lymphokine production by mononuclear cells (PBMC), lymphocyte subpopulations, and natural killer cell (NK) activity were measured at baseline and 4 months of follow-up (before vaccination). Antibodies against influenza and pneumococcal antigens and flu-stimulated production of interferon gamma and interleukin-2 by PBMC were measured at 4 and 6 months. Skin response to 7 recall antigens and body composition were assessed at baseline and at 4 and 12 months. All infections occurring during the study period were recorded.NK activity increased in supplemented subjects and decreased in nonsupplemented individuals. Interleukin-2 production by PBMC and the proportion of T cells with NK activity decreased in controls and did not change in supplemented subjects. Supplemented subjects reported less infections than nonsupplemented individuals (in 13% and 22% of scheduled visits, respectively; p = .02).This nutritional supplement increased innate immunity and protection against infections in elderly people.

The mechanism by which homocysteine (Hcy) causes endothelial dysfunction is probably mediated by oxidative stress. The aim of this study was to evaluate the effect of oxidative stress on endothelial function in young and elderly hyperhomocysteinemic (HHcy) men. A total of 35 HHcy (Hcy > 15 micro mol/L), young (n = 15; 20-40 y) and elderly men (n = 20; > 65 y) and 33 normohomocysteinemic (NHcy; controls) young (n = 14) and elderly (n = 19) men (Hcy < 13 micro mol/L), without classic cardiovascular risk factors were recruited. Serum Hcy, folate, and vitamin B-12, whole-blood glutathione, plasma total antioxidants status, TBARS, and 8-F(2alpha) isoprostanes were determined. Noninvasive ultrasound measurements of endothelium-dependent (EDVR) and -independent dilatation (EIVR) were performed. EDVR, EIVR, and markers of oxidative stress did not differ among the groups. Folate concentrations were higher in elderly than in young men (P < 0.001), independent of Hcy concentrations. Vitamin B-12 concentrations were lower in HHcy than in NHcy elderly men (P < 0.045). EDVR was correlated with folate concentrations in young men (r = 0.40, P = 0.04) and negatively with BMI in elderly men (r = -0.52, P = 0.002). In the present study, HHcy with normal serum folate concentrations was not associated with poor EDVR or oxidative stress in healthy young and elderly men.

Objective There is a seasonal variation in the incidence of diabetes mellitus and cardiovascular diseases. However, there is very little information about the seasonal variation in insulin sensitivity. We report the seasonal variation in insulin sensitivity in a group of elderly subjects followed for 1 y. Methods Healthy elderly (≥70 y) subjects living independently were included. Fifty percent of subjects received a daily nutritional supplement that provided 400 kcal, 15 g of protein, and 50% of vitamin daily reference values (DRVs). Those receiving and not receiving supplements were randomly assigned to a resistance exercise training program. Every 6 mo (winter, summer, and winter), body composition was measured by dual-energy x-ray absorptiometry and blood samples were used to measure serum lipids, fasting and postprandial glucose, and insulin levels. Results One hundred eight subjects (31 supplemented and trained, 28 supplemented, 16 trained, and 33 without supplementation or training) completed the follow up. Higher homeostasis assessment of insulin sensitivity, postprandial insulin, and fasting triacylglycerol levels were observed during the summer than during the winter. Body fat increased steadily during the study period, and fat-free mass did not change. Serum low-density lipoprotein cholesterol decreased significantly in the supplemented and trained group and increased in the non-intervention group. Conclusions In this group of elderly subjects, insulin resistance and triacylglycerol levels were higher during the summer. Nutritional supplementation and training had a positive effect on serum low-density lipoprotein cholesterol.

Background: The relationship between hyperhomocysteinemia and cardiovascular disease has not been totally elucidated. Hypothesis: The study aimed to verify the association between hyperhomocysteinemia and endothelial dysfunction before and after modification of total homocysteine (tHcy) serum levels with vitamin supplementation in young male subjects devoid of any other cardiovascular risk factor. Methods: Twenty hyperhomocysteinemic (tHcy gt; 15 μmol/1) male volunteers (≤ 40 years) and 20 age-matched subjects with normal tHcy levels (tHcy < 13 μmol/1) were included. Exclusion criteria were smoking, hypertension, diabetes, vitamin ingestion, obesity, hypercholesterolemia, renal failure, and positive antiphospholipid antibodies. Serum tHcy, folate, vitamin B12 levels, activated protein C and S, protein C resistance, fibrinogen, prothrombin, thrombin, antithrombin III, and in vitro oxidation of low-density lipoprotein (LDL) particles were measured. Noninvasive ultrasound measurements of endothelium-dependent (EDD) and -independent dilatation (EID) of the brachial artery were performed. Each pair was then randomly assigned to receive a vitamin capsule (0.6 mg folic acid, 0.8 mg B12, and 2.0 mg B6) or an identical placebo during 8 weeks, in a double-blind study design. After the treatment phase, blood samples and vascular reactivity were repeated. Results: Nine pairs of volunteers received vitamins and 11 received placebo. Hyperhomocysteinemic subjects had lower baseline serum levels of vitamin B12. Serum folate levels, antithrombotic function, in vitro LDL oxidation, and EDD were similar in all groups. After the vitamin supplementation, serum folic acid levels increased significantly both in normo-and hyperhomocysteinemic subjects, unlike vitamin B12, which increased only in the hyperhomocysteinemic individuals. Plasma tHcy decreased significantly in the supplemented groups. Treatment with vitamins was not associated with improvement in EDD or antithrombotic function. Conclusions: Mild hyperhomocysteinemia is not associated with endothelial dysfunction in young male subjects with no additional cardiovascular risk factors, and reduction of tHcy by vitamin supplementation does not modify EDD in this age group. In this sample, tHcy was more related to vitamin B12 than to folic acid status.

Prebiotics stimulate the growth of bifidogenic bacteria in the gut. The aim of this work was to assess the effects of a prebiotic mixture on the immune response in healthy elderly people.Healthy free-living elderly people (age,or = 70 years), receiving a nutritional supplement that provided 1.6 MJ, 15 g of protein, and 50% of vitamin daily reference values per day, were randomly assigned to receive a prebiotic mixture (6 g/d of a 70% raftilose and 30% raftiline mixture) or placebo (6 g of maltodextrin powder) for 28 weeks. At week 2 of the study, all subjects were vaccinated with influenza and pneumococcal vaccines. At weeks 0, 2, and 8 of the study, serum total proteins, albumin, immunoglobulins, saliva secretory immunoglobulin A (IgA), and serum titers of influenza A and B and pneumococcal antibodies were measured. At week 8, cultured peripheral monocyte cell secretion of interleukin-4, interferon-gamma, and lymphocyte proliferation, stimulated with phytohemagglutinin and influenza antigen, were measured.Sixty-six subjects were considered eligible for the study, and 43 (20 receiving prebiotics and 23 receiving placebo) were considered for final analyses on a per protocol basis. No changes in serum proteins, albumin, immunoglobulins, and secretory IgA were observed. Antibodies against influenza B and pneumococcus increased significantly from weeks 0 to 8, with no significant differences between groups. Antibodies against influenza A did not increase. No effects of prebiotics on interleukin-4 and interferon-gamma secretion by cultured monocytes were observed.No immunological effects of prebiotics were observed in this study.

OBJECTIVES: To compare endothelium-dependent vasomotor response in healthy younger and older subjects without classic cardiovascular risk factors, with high and normal fasting homocysteine (tHcy) levels. DESIGN: We compared endothelium-dependent vasodilatation, using ultrasound, in healthy younger (aged 18–40) and older (≥70) people with normal ( 15 μmol/L) tHcy levels. Exclusion criteria were smoking, personal history of cardiovascular disease, hypertension, chronic diseases, vitamin intake, obesity, abnormal serum lipids levels, and creatinine higher than 130 μmol/L. SETTING: Research laboratory. MEASUREMENTS: In addition to tHcy levels, serum folate and vitamin B12 levels were measured. RESULTS: We studied 17 younger and 12 older hyperhomocysteinemic subjects and respective aged-matched normohocysteinemic subjects. Endothelium-dependent vasodilatation was lower in the hyperhomocysteinemic older people (P < .01) than in all younger subjects and in normohomocysteinemic older people. Serum vitamin B12 levels were higher in younger and older normal controls. Folic acid levels were higher in younger controls and in both older groups. CONCLUSIONS: This study shows an effect of high circulating tHcy on vascular reactivity in older people. Because serum levels of tHcy are associated with nutritional status of vitamin B12 and folic acid, prospective studies are necessary to demonstrate the effects of a long-term nutritional supplementation with vitamins on vascular function and global cardiovascular risk.

In the placenta, 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) limits fetal glucocorticoid exposure and its inhibition has been associated to low birth weight. Its expression, encoded by the HSD11B2 gene is regulated by DNA methylation. We hypothesized that maternal diets supplemented with folic acid (FA) during pregnancy modify the expression of placental HSD11B2 through gene methylation. Wistar rats were fed with high (8 mg/kg) or normal low (1mg/kg, control) levels of FA during pregnancy. Concentrations of mRNA and protein in placentas were determined by qRT-PCR and Western blot respectively. Methylation in five CpG sites of the placental HSD11B2 promoter (−378 to −275) was analyzed by bacterial cloning and subsequent sequencing. In the FA-supplemented group, mRNA and protein levels of 11β-HSD2 decreased by 58% and increased by 89%, respectively, only in placentas attached to males. In controls, most CpG sites were not methylated except for the CpG2 site which was 80% methylated. CpG2 methylation level increased under the FA treatment; however, only in placentas attached to females was this increase significant (113%). This change was not related to HSD11B2 expression. Fetal weight of females from FA- supplemented mothers was 6% higher than females from control mothers. In conclusion, this is the first study reporting that FA over supplementation during pregnancy modifies the placental HSD11B2 gene expression and methylation in a sex-dependent manner, suggesting that maternal diets with high content of FA can induce early sex-specific responses, which may lead to long-term consequences for the offspring.

Background Simple and reliable methods to evaluate muscle mass in older people are lacking. Aim To evaluate ultrasound as a measure of RF muscle mass and quality in healthy subjects of different ages and gender, assessing its concordance with dual energy X-ray densitometry (DEXA) and association with muscle strength and walking capacity. Methods We selected 54 adults of both genders, aged 20–55 years and 51 adults older than 60 years. Ultrasound images of the RF were obtained at the mid-thigh to measure its thickness and ultrasonographic density using a GE Logiq e equipment. Body composition was assessed by DEXA. Quadriceps isometric strength and 12 minutes’ walk were also measured and gender specific t scores for older adults were calculated using the values obtained in adults. Results RF ultrasound measurements correlated significantly with lean body mass assessed by DEXA (Double energy X-ray absorptiometry). The concordance between both measures was also adequate. Older people had lower muscle mass and worse ultrasound parameters than adults. Older males with a t score for quadriceps strength of −2 or less, had a significantly higher RF grayscale density. Older males with a 12 minutes’ walk t score of −2 or less and old males and females with a walking speed of 1 m/s or less had a lower RF thickness. Discussion There is a good concordance between RF ultrasound and DEXA. Conclusions Assessment of RF using ultrasound appears to be a reliable and accurate method to evaluate muscle mass in older people.

To explore the usefulness of carotid ultrasound examination as a marker of aging and predictor of disability among older people.Carotid ultrasound, measuring carotid intima media thickness (CIMT) and recording the presence of plaques, was carried out in 152 adults aged 29-59 years (47 women) and in 107 older adults aged 61-88 years (86 women). In all, clinical routine laboratory parameters and lymphocyte telomere length as T/S ratio were measured. Among older adults, 12-min walk, timed up and go, hand grip and quadriceps strength were determined.CIMT was significantly higher among older people and T/S ratio was significantly higher in young women. Carotid plaques were found in one adult and 17 older people. A multiple regression analysis accepted age, systolic blood pressure and T/S ratios as independent predictors of CIMT (R(2) = 0.51). Among older people, a logistic regression accepted age and the presence of carotid plaques as significant predictors of a 12-min walk speed below 1 m/s.An abnormal 12-min walk as an indicator of functional decline among older people is associated with the presence of carotid artery plaques. CIMT is independently associated with age.

To test the variability within and between subject of glycemic response test following the ingestion of a standard food.Glucose and insulin response of a standard meal (white bread) was performed in ten healthy volunteers and repeated under identical conditions for 6 times. Blood glucose and insulin levels were measured in the fasted state and over the 180 min following commencement of consumption of the foods The Area Under the Curve (AUC) for glucose and insulin was calculated for the values above baseline for the 3-hour period following the standard meal. Within and between coefficient of variation was calculated.The total intra-individual variation of the gAUC was 51.8% range 24.9 to 91.4%. The inter-individual variation of the gAUC in the complete study was 75.2% . The total intra-individual variation of the iAUC was 51.9%. ranged: 7.7 to 103%. The inter-individual variation in the complete study was 86%.Glucose and insulin response to a reference food has low reliability, therefore limits its clinical utility for individual dietary prescription.Objetivo: Evaluar la variabilidad inter e intra individual de la respuesta glicémica frente a la ingestión de un alimento estándar en sujetos sanos. Material y métodos: Se midió en 6 oportunidades la respuesta glicémica e insulínica de un alimento estándar (pan batido blanco) en 10 voluntarios sanos. A cada sujeto se le midió la glicemia e insulina en ayuno a los 15, 45, 60, 90, 120, 150 y 180 minutos, después de ingerir el alimento estándar. Posteriormente se calculó el área bajo la curva (ABC) de glicemia e insulina en cada sujeto. Se calculó el coeficiente de variación intra e inter individual del ABC de glucosa e insulina. Resultados: La variación intra-individual total del ABC de glucosa fue de 51,8% (rango: 24,9%-91,4%). La variación inter-individual del ABC de glucosa total fue 75,2%. La variación intra-individual del ABC de insulina fue de 51,9% (rango: 7,7%-103%). La variación inter-individual del ABC de insulina total fue de 86%. Conclusión: La respuesta glicémica e insulínica a un alimento estándar tiene poca reproducibilidad, tanto intra e inter individual. En consecuencia, su utilidad clínica para la prescripción dietaria individual, está limitada.

The Chilean Ministry of Health developed a healthy lifestyles intervention directed to adults with overweight and cardiovascular risk factors, called "Program on Healthy Eating and Physical Activity" (PASAF).To evaluate the impact of PASAF on nutritional status and metabolic parameters.We analyzed databases from three primary care centers belonging to a municipality of Metropolitan Santiago. We selected adults enrolled in the PASAF during three years (2007-2009). The program lasted four months and included an assessment of anthropometric and metabolic parameters at baseline and at the end, eight workshops with a nutritionist, seven with a psychologist and 32 sessions of physical activity.We evaluated 526 subjects aged ≥18 years (93% females), of whom 85.6% attended the last appointment for assessment. Analyzing available data, attendance to workshops was50% of the scheduled sessions. Weight, body mass index and waist circumference decreased significantly (median: -1.4 kg, -0.6 kg/m² and -3 cm, respectively). The median weight loss was 1.8% of initial weight and 17.1% of participants experienced a decrease ≥5% of their initial weight. There were significant improvements in lipid levels and blood pressure among participants with lower initial excess weight. A reduction in fasting blood glucose was observed only among subjects who lost ≥5% of their initial weight.The PASAF modestly reduced nutritional parameters. Correction of metabolic parameters was especially effective in less obese subjects. The attendance to workshops was low.

Alcohol ingestion may promote lipid peroxidation, and the presence of polyunsaturated fatty acids in liver lipids may be essential for the generation of liver damage through this mechanism. The aim of this study is to examine fatty acid composition of liver lipids in chronic alcoholics with and without histological liver damage. A percutaneous liver biopsy was performed to 28 patients hospitalized for treatment of their alcoholism. Liver total lipids were extracted from a portion of the tissue sample and fatty acid composition was measured by gas chromatography. Another piece of the sample was sent for histological study. Six patients had histological cirrhosis or alcoholic hepatitis in their biopsies, the rest of the patients had minimal changes. Patients with liver damage had higher levels of oleic acid and total monoenoic fatty acids, a higher 18:1/18:0 ratio, lower levels of polyunsaturated fatty acids, a lower 20:4/18:2 ratio, and a lower peroxidability index in liver total lipids, than patients without liver damage. Alcoholic patients with asymptomatic liver damage have less unsaturated fatty acids in liver total lipids than their counterparts with normal livers.

Excessive ethanol ingestion induces hypoandrogenism in male subjects. To confirm its presence and to study its relationship with the degree of liver damage and alcohol abstinence, plasma sex hormones were measured in alcoholic patients without liver failure, after two different abstinence periods. Patients were 30 male chronic alcoholics admitted to the Alcoholism Ward for treatment of their addiction. On admission, we measured: testosterone (T), estradiol (E), follicle stimulating hormone (FSH), luteinizing hormone (LH) and sex-hormone binding globulin (SHBG). A liver biopsy was also performed. These measurements were repeated at discharge and were also done in 15 normal volunteers. On admission (mean abstinence 1.9 +/- 1.7 days) total T was similar to controls, FSH was lower (p0.02) and high levels of SHBG were found (3.5 fold increase, as compared to controls). Histologically, 9 patients had normal liver; 14 had moderate alterations and 7 showed marked alterations. Hormonal values were not different in these 3 groups. At discharge, 11.1 +/- 4.7 days after admission, T, E and FSH did not show significant changes but LH decreased (8.2 +/- 5.2 mIU/ml vs 12.9 +/- 4.1, p0.001); SHBG also decreased (65.4 +/- 21.6 nmol/l vs 117.2 +/- 33.3, p0.001) to values that still were twice those of controls. It is concluded that alcoholic patients without clinical signs of liver failure have normal plasma testosterone levels, irrespective of their histologic liver alterations and high plasma SHBG levels that decreased significantly after a short abstinence. The concomitant LH decrease suggests that hypoandrogenism is likely in these patients. Fast changes in SHBG levels rise the possibility that this protein is candidate marker of alcoholism.

Alcohol ingestion promotes lipoperoxidation and alters cellular antioxidant mechanisms. Alpha-tocopherol levels decrease in alcoholics as severity of liver damage increases. The aim of this protocol was to study the effects of a long-term oral 500 mg vitamin E daily supplementation in decompensated ambulatory alcoholic cirrhotics.67 subjects were included in this double blind trial; 33 patients received vitamin E and 34 patients received placebo tablets of identical appearance during 1 year. Each month, the patients were seen by a nurse practitioner who was in charge of detecting alcohol ingestion and checking adherence to treatment. Every 3 months, the patients underwent a medical examination, and blood samples were taken for clinical laboratory analysis and serum vitamin E measurement.Alpha-tocopherol levels were significantly lower in patients with more severe liver disease. This difference was not significant when vitamin E levels were corrected by cholesterol. Oral supplementation significantly increased serum vitamin E levels in the experimental group. Alcohol ingestion and hospitalization rates were similar in both groups. Life table analysis did not show significant differences in mortality between the two groups.Vitamin E supplementation with adequate doses of an alpha-tocopheryl acetate formulation during 1 year did not influence hepatic laboratory parameters, mortality or hospitalization rates of decompensated alcoholic cirrhotics, although serum levels of the vitamin significantly increased.

Objectives Folate supplementation may be associated with an increased risk of developing several types of cancer and a derangement of immune function. Among the latter, Natural killer (NK) cells are involved in non–MHC-restricted natural immunity against malignant target cells. Abnormalities in NK cell number or function have been associated with a higher cancer risk. The aim of this study was to study in vitro the possible effect of different concentrations of 5-methyltetrahydrofolic acid (5-MTHF) or folic acid on NK cell cytotoxic function, and expression of the stimulatory and inhibitory receptors KIRDL4, KIRDL3, and NKG2D. Methods Volunteer-derived peripheral mononuclear cells (PBMC) and highly enriched NK cells (95% CD56+ CD16+) were grown in folic acid free–RPMI 1640, supplemented either with folic acid or 5-MTHF (15–100 nM) during 72 h to 96 h. Results No differences in the cytolytic activity of PBMC and enriched NK cells were observed. After 96 h of in vitro culture without folate or supplemented with FA or 5-MTHF (30 or 100 nM), there were no changes in the percentage of HPNK receptor-positive cells. Conclusions Our data indicate that a high dose of 5-MTHF or folic acid does not influence NK cell function in vitro.

Exercise training during the dialytical procedure may have positive cardiovascular effects and prevent or revert muscle wasting in patients undergoing chronic hemodialysis.To evaluate the effects of an exercise training program in patients undergoing chronic hemodialysis.Fifteen patients on chronic hemodialysis aged 21 to 69 years (three females) were included in the study. Nine of these were included in an exercise training program. During 16 weeks, exercise sessions were carried out during each dialytical procedure that included a warm-up period, aerobic exercises done using standing cycles, and resistance exercises, performed using Thera-Band(®) elastic bands and loops. Borg scale was used to control the intensity of training. At baseline and at the end of the study, a blood sample prior and after the dialytical procedure was obtained to measure C reactive protein, tumor necrosis factor α and interleukin 6. Quadriceps muscle strength, six minutes' walk and quality of life using the SF-36 questionnaire, were also measured.Four experimental subjects did not complete the study period, two that withdrew before starting, one due to problems with the venous access and one that decided to withdraw after 1 month of training. Among the five patients that finished the training period, significant improvements in the six minutes' walk and quadriceps strength were observed in the experimental group. No significant changes were observed among controls. No changes were observed in either group in C reactive protein, tumor necrosis factor and interleukin 6 levels or quality of life.Among patients undergoing chronic hemodialysis exercise training improves endurance and muscle strength.

Thirteen asymptomatic chronic alcoholic patients were studied to investigate the early stages of portal hypertension in alcoholic liver disease and the effects of withdrawal and ethanol on hepatic function and hemodynamic variables. None of the patients presented clinical signs of decompensated liver disease, and their liver biopsies showed normal liver or moderate alterations only. In basal conditions and after an intravenous ethanol infusion (1 g/kg body weight), hepatic venous pressure gradient and hepatic blood flow using indocyanine green were measured through hepatic vein catheterization. Hepatic sinusoidal vascular resistance and indocyanine green intrinsic clearance were also calculated. Portal blood flow measurements were obtained by Doppler ultrasound. No correlation was observed between hepatic venous pressure gradient and histologic features, (steatosis, necrosis, fibrosis, inflammation and hepatocyte surface area). In basal conditions, portal hypertension was not found in any case. After ethanol, portal pressure increased significantly (p0.001); in four cases it rose to or above 5 mmHg. Portal blood flow, hepatic blood flow and hepatic vascular resistance also increased significantly. Intrinsic indocyanine green clearance decreased slightly but significantly. No significant correlations were found between portal pressure, hepatic resistance and the histologic parameters. It was concluded that alcoholic patients, without clinical or laboratory evidence of liver failure and with minimal or moderate histologic alterations, have normal portal pressures. After an intravenous ethanol load, however, four out of 13 patients (31%) reached levels of 5 mmHg or more, irrespective of their liver histology.

To evaluate the association between loss of fat-free mass and mortality among older people.Information of healthy Chilean older people evaluated by dual energy X-ray absorptiometry was used, identifying those who died in a period of 12 years. A Cox proportional hazards model was used to identify mortality predictors. Life tables were constructed calculating survival using predictive variables.Information from 1413 participants aged 74.3 ± 5.6 years (1001 women), was obtained. During the follow-up (median 1594 days), 221 participants died. The Cox model identified age and appendicular fat-free mass as predictors of death (hazard ratios 1.08 and 0.85, respectively). According to life tables, participants in the lower sex-specific quartile for appendicular fat-free mass/height had significantly higher mortality. This association was significant among participants aged over 73 years.A low fat-free mass was predictive of mortality in older people.

Human adipocyte precursor cells (APC) have been characterized in their proliferation and differentiation potential from subcutaneous, omental, and mesenteric depots, mostly from morbidly obese patients. Cells from the preperitoneal adipose compartment have not been characterized yet, least of all when obtained from normal weight subjects. The aim was to compare proliferation and differentiation of subcutaneous (SC) and preperitoneal (PP) APC derived from adipose tissue in healthy subjects with different body mass. SC and PP adipose tissue was obtained during surgery of inguinal hernias in five healthy non-obese subjects and three obese otherwise healthy men. APC, obtained by collagenase digestion, were cultured. Proliferation was assayed by cell counting and differentiation by oil red O staining and flow cytometry using Nile Red staining. Proliferation of SC was higher than PP APC. Such differences between both compartments were even higher in APC obtained from obese patients. Conversely PP APC differentiated earlier in vitro compared with SC cells. These results agree with published data on fat cell proliferation. However regarding differentiation, our data show that APC from deeper depots (in this case PP) differentiate earlier than subcutaneous APC. This is different to previous studies performed in mesenteric or omental adipose tissue.

Antibiotic-associated diarrhea is one of the most common adverse effects of antimicrobials. Any antimicrobial can potentially produce diarrhea but beta-lactamics have a higher risk. Among these, amoxicillin is widely indicated in ambulatory practice. One of the alternatives suggested to prevent antibiotic-associated diarrhea, is the use of the probiotic Saccharomyces boulardii.To evaluate whether the concomitant use of Saccharomyces boulardii and amoxicillin can prevent antibiotic associated diarrhea in ambulatory adults with acute infections diseases, without provoking other adverse effects.Eighty six adults (aged 15 to 81 years) with acute infectious diseases, excluding those arising in the gastrointestinal tract, that received a prescription of oral amoxicillin for 5 to 10 days, were included. In a controlled randomized, double blind trial, 41 patients were assigned to receive lyophilized Saccharomyces boulardii (500 mg/day) during 12 days, and 45 patients were assigned to placebo for the same period.Ten percent of patients (9/86) reported acute diarrhea, 9,8% (4/41) in the experimental group and 11.196 (5/45) in the control group (p = 100). No adverse effects were associated to the use of the probiotic.Saccharomyces boulardii (500 mg/day) did not prevent diarrhea related to amoxicillin.

We assessed the impact of high serum folate concentration on erythrocyte S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH) concentrations, SAM/SAH ratio, CpG methylation levels across the promoter region of the extracellular superoxide dismutase (ec-SOD) gene, and ec-SOD activity in healthy men.Serum folate levels were measured in 111 subjects who were categorized in quintiles according to their folate status. Subjects located at the lowest, middle, and upper quintiles were selected for assessment of SAM and SAH by high-performance liquid chromatography, C677T genotype of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene, ec-SOD methylation of CpG sites in lymphocytes genomic DNA by bisulfate treatment, and ec-SOD activity by a chemical assay.Sixteen subjects were in the lowest serum folate quintile (23.6 nmol/L), 17 in the middle (34-42 nmol/L), and 14 in the highest (45nmol/L). SAM concentration was higher in the upper than in the middle and lowest quintiles (5.57 +/- 1.58, 2.52 +/- 0.97, 2.29 +/- 1.2 micromol/L; P0.0001). SAH concentration was higher in the upper compared with the lowest quintile (0.76 +/- 0.24 versus 0.52 +/- 0.23 micromol/L, P0.001). There were no differences in the SAM/SAH ratio, ec-SOD activity, methylation status of CpG sites of the ec-SOD gene, and TMTHFR C677T genotype between groups.Serum folate concentrations in the highest quintile among healthy humans are associated with increased erythrocyte SAM and SAH concentrations, but not with SAM/SAH ratio or with methylation levels of CpG sites across the promoter region of the ec-SOD gene. Further research is required to determine if these findings are beneficial or harmful.

Summary Background Endothelial dysfunction is a key process in atherosclerosis. Hypomethylation is one of the postulated mechanisms involved in atherogenesis and is mainly secondary to a decrease in essential factors such as, folate and vitamin B12 for the biosynthesis of S -adenosylmethionine (SAM), the main methyl-group donor for methylation reactions. Aim To investigate in an animal model, whether hypomethylation, secondary to folate or vitamin B12 deficiency, affects endothelium-dependent relaxation (EDR) induced by acetylcholine (ACh). Methods Adult male Wistar rats were divided into 4 groups of 12 rats each: folate and B12 deficiency (FB12D 0mg folate/kg, 0μg/kg B12), folate deficiency (FD 0mg folate/kg and 50μg/kg B12), B12 deficiency (B12D: 8mg/kg folate and 0μg/kg B12 and control diet (CD)). After eight weeks the animals were killed and thoracic aorta and liver removed. Serum concentration of homocysteine, folate and vitamin B12 were determined. Hepatic levels of SAM and S -adenosylhomocysteine (SAH) were measured, as indicator of hypomethylation. ACh-induced EDR and sodium nitroprusside (SNP)-induced endothelium-independent relaxation (EIR), in isolated aorta rings were evaluated. Results Hcy concentrations were significantly increased in the folate and B12 deficient groups. SAM and the SAM/SAH ratio were lower in the FD and FB12D than in the control and B12D group. Folate, B12 deficiency, serum Hcy levels and hepatic SAM/SAH ratio did not affect EDR neither EIR. Conclusions In adult Wistar rats, chronic folate or folate plus vitamin B12 deficiency generates hypomethylation which is not related to an alteration of endothelial function.

In acute illnesses, plasma glucose levels are often increased and generally parallel the severity of stress. Hyperglycemia caused by reduced insulin sensitivity and reduced insulin secretion is associated with increased susceptibility to infections. Maintaining blood glucose levels at or below 110 mg/dl reduces morbidity and mortality in critically ill patients.To measure the glucose and insulin responses of four commercially available enteral formulas compared with a standard meal reference product.The glycemic index (GI) and the insulin index (II) were determined in a randomized, cross over protocol in 38 healthy volunteers between 18 and 46 years of age. Each subject underwent five tests: three with the standard meal (bread) and two with the study products. The enteral formulas were Clinutren HPR (whole protein of high protein value), Crucial (casein peptide based formula), Peptamen, (whey peptide based formula), Glytrol (formula for diabetics with whole protein with fiber). Each study product was evaluated 10 times.The diabetic formula and the high protein energy dense formulas induced a significantly lower GI (p0.02) compared with the standard meal. The GI response did not appear to be due to enhanced insulin secretion. The other tested formulas had lower GI than the standard meal, but in addition they exhibited increased II The whey based peptide formulation produced the highest insulin response (p0.03).Both GI and II are related to the concentration, form and type of protein contained in the enteral formula. The whey peptide formulation produced a low GI with the highest insulin index. Based on the low GI of these enteral products, all can be useful to provide nutritional support during metabolic stress, without adding an additional challenge to blood glucose management.

Background and aims: An accurate diagnosis of sarcopenia is required. The aim of this study is to correlate the results of two methods to define sarcopenia using cross sectional body composition data, with actual loss of fat free mass. Methods: Healthy older subjects (926 females and 381 males aged 70 years or more) and healthy young adults (425 females and 151 males aged 20 to 40 years) were studied. Body composition was assessed by double beam X ray absorptiometry (DEXA). Among older subjects, a contemporary subsample of 148 females and 45 males had two or more measurements, separated by 4.8±1.5 years and loss of fat free mass per year was calculated. In the whole sample, total and appendicular lean body mass index were calculated as total or appendicular lean body mass/height. Using data from young people, sex specific t scores were obtained. In older subjects residuals were derived from a regression equation, using total or appendicular fat free mass as the dependent variable and height, fat free mass and age as independent variables. Results: The concordance between residuals and t scores to define sarcopenia was 68 and 72%, respectively. Among subjects with two or more measurements, men and women lost a mean of 521±454 and 221±399 g/year of fat free mass, respectively. The odds ratio of losing more than 822 g lean body mass/year among men or 514 g lean body mass/year among women was 2.63 and 2.64 for subjects classified in the two lowest quintiles of sarcopenia, using t scores or residuals, respectively. Conclusions: Cross sectional body composition data can predict loss of fat free mass among older people.

Introduction: Advanced glycoxidation end-products (AGEs) are involved in age-related conditions and diabetic complications. Diet intake contributes to their circulating concentrations. Aim: To measure serum and urinary AGEs in non-diabetic volunteers and relate their concentration to body composition, blood chemistry and dietary ingestion. Methods: We studied 41 adult men (31 middle-aged adults and 10 elderly). A nutritional assessment including a dietary recall designed for detection of AGE ingestion (specifically carboxymethyl-lysine(CML)), and anthropometric measurements were performed. Also serum lipoproteins, insulin, glucose, leptin and C reactive protein (CRP). AGEs were measured in serum and urine samples using size exclusion chromatography and flow injection assay (FIA); the technical procedures were first employed in 11 heterogeneous diabetics, as positive controls for this methodology. Results: Serum and urinary chromatograms indicated that areas under the curve were not different in younger compared with elderly adults. AGEs did not correlate with dietary intake, body composition, nor metabolic parameters, however they correlated significantly with renal function and CRP concentration. Discussion: In these non-diabetic volunteers, with low CML intake, serum and urinary concentration of AGEs were not related to dietary intake. AGEs were related to renal function and CRP, but not to body composition, lipoproteins, insulin and glucose. Key terms: Advanced glycation end-products, Fluorescent AGEs, glycoxidation.

To study the effects of a special nutritional supplement on bone mineral density and bone turnover markers in Chilean elderly subjects with femoral osteoporosis.Public primary health care clinics in Chile.Free living elderly subjects with femoral osteoporosis.Subjects were randomized to receive the usual nutritional supplement provided by the Chilean Ministry of Health or a special nutritional supplement providing, among other nutrients, 90 mg isoflavones, 800 mg calcium, 400 IU vitamin D, 60 ug vitamin K and 31 g proteins per day.At baseline, and after six and twelve months of supplementation, body composition, bone mineral density, serum 25 OH vitamin D, intact parathyroid hormone (iPTH), osteocalcin, decarboxylated osteocalcin, urinary aminoterminal telopeptide of type I collagen (NTX), deoxypyridoline cross links (Dpd) and equol were measured. Every month, urinary daidzein was measured in a morning urine sample.No differences between treatment groups were observed in body composition or bone mineral density changes. The group receiving the special supplement had a significant increase in serum 25 OH vitamin D and a significant decrease in serum iPTH and decarboxylated osteocalcin. No association between daidzein or equol excretion and changes in bone mineralization was observed.A special supplement delivered to elderly subjects with osteoporosis improved serum vitamin D and reduced serum iPTH and undercarboxylated osteocalcin levels but did not affect BMD.