Re-mapping the molecular features of the human immunodeficiency virus type 1 and human T-cell lymphotropic virus type 1 Brazilian sequences using a bioinformatics unit established in Salvador, Bahia, Brazil, to give support to the viral epidemiology studies

The analysis of genetic data for human immunodeficiency virus type 1 (HIV-1) and human T-cell lymphotropic virus type 1 (HTLV-1) is essential to improve treatment and public health strategies as well as to select strains for vaccine programs. However, the analysis of large quantities of genetic data requires collaborative efforts in bioinformatics, computer biology, molecular biology, evolution, and medical science. The objective of this study was to review and improve the molecular epidemiology of HIV-1 and HTLV-1 viruses isolated in Brazil using bioinformatic tools available in the Laboratório Avançado de Sáude Pública (Lasp) bioinformatics unit. The analysis of HIV-1 isolates confirmed a heterogeneous distribution of the viral genotypes circulating in the country. The Brazilian HIV-1 epidemic is characterized by the presence of multiple subtypes (B, F1, C) and B/F1 recombinant virus while, on the other hand, most of the HTLV-1 sequences were classified as Transcontinental subgroup of the Cosmopolitan subtype. Despite the high variation among HIV-1 subtypes, protein glycosylation and phosphorylation domains were conserved in the pol, gag, and env genes of the Brazilian HIV-1 strains suggesting constraints in the HIV-1 evolution process. As expected, the functional protein sites were highly conservative in the HTLV-1 env gene sequences. Furthermore, the presence of these functional sites in HIV-1 and HTLV-1 strains could help in the development of vaccines that pre-empt the viral escape process.

Cellular origin of T-cell lymphomas.

In this issue of Blood, Iqbal et al, having compiled gene expression profiles from >300 peripheral T-cell lymphomas, expand previous findings on the diagnostic value of molecular signatures that correlate with different histological types of T-cell lymphomas. They report the discovery of 2 molecular subgroups of peripheral T-cell lymphomas, not otherwise specified (PTCL, NOS), characterized by high expression of either GATA-binding protein 3 (GATA-3) or t-box 21 (TBX21) transcription factors and corresponding target genes, with the GATA3 subgroup being associated with distinctly worse prognosis. In an independent study, Wang et al(2) also show that GATA3 expression in a subset of PTCL, NOS identifies a subgroup of patients with inferior survival.

Metabolic effects of fructose in humans

Abstract : Fructose is a simple sugar, whose consumption has increased over the past decades. In rodents, a high-fructose diet (HFrD) induces several features of the metabolic syndrome. The aim of the studies included in this thesis was to investigate the metabolic effects of a HFrD in humans, with a focus on insulin sensitivity and ectopic fat deposition. Moreover, we addressed the question whether these effects may differ between individuals according to gender and the genetic background. The first study was designed to evaluate the impact of a 4-week HFrD on insulin sensitivity and lipid metabolism in 7 healthy men. Insulin sensitivity, intrahepatocellular lipids (IHCL) and intramyocellular lipids (IMCL) contents were measured before and after 1 and 4 weeks of HFrD (1.5 g fructose/kg body weight/day). Insulin sensitivity was assessed by a 2-step hyperinsulinemic euglycemic clamp. IHCL and IMCL were measured by 1H-magnetic resonance spectroscopy (MRS). Fructose caused significant (P<0.05) increases in fasting plasma concentrations of triacylglycerol (TG) (+36%), VLDL-TG (+72%) and glucose (+6%) without any change in body weight, IHCL, IMCL, and insulin sensitivity. In the second study, muscle biopsies were taken from five of these healthy male subjects before and after 4 weeks of HFrD. mRNA concentrations of 18 genes involved in lipid and carbohydrate metabolism were quantified by real-time quantitative PCR. We found that a 4-week HFrD increased the expression of genes involved in lipid synthesis, while it decreased those involved in insulin sensitivity and lipid oxidation; these molecular changes maybe early markers of insulin resistance and altered lipid metabolism. The third study aimed at delineating whether male and females equally respond to a HFrD. For this purpose, higher doses of fructose (twice the dose of the previous study) were provided to 8 healthy young males and 8 healthy young females over 6 days. HFrD significantly increased fasting TG in males (+71 %), whereas this increase was markedly blunted in females (+16%). Males also developed hepatic insulin resistance, characterized by increased hepatic glucose output (+12%), and showed higher alanine aminotransferase concentration (+38%), but none of these effect was observed in females. This study suggests that short-term HFrD leads to hypertriglyceridemia and hepatic insulin resistance in men, but premenopausal women seem protected against these effects. Finally, the fourth study investigated whether healthy offspring of type 2 diabetic patients (OffT2D), a subgroup of individuals prone to metabolic disorders due to their genetic background, may have exacerbated response to HFrD. Eight healthy males (Ctrl) and 16 OffT2D received a HFrD and isocaloric diet in a randomized order. In both groups, HFrD significantly increased IHCL (Ctrl: +76%; OffT2D: +79%) and fasting plasma VLDL-TG (Ctrl: +51 %; OffT2D: +110%). In absolute values, these increments were significantly higher in OffT2D, suggesting that these individuals may be more prone to developing metabolic disorders when challenged by high fructose intake. In order to better delineate the specific effects of fructose vs the hypercaloric energy content, we repeated the complete metabolic investigations after an isocaloric high glucose diet in four of the eight Ctrl volunteers. After a high glucose diet, TG and IHCL concentrations remained similar to the control values, in contrast to the marked increases observed after the HFrD. In conclusion, the studies included in this thesis provided novel insights into the metabolic effects of fructose in humans. They showed that fructose may rapidly increase fasting VLDL-TG, IHCL and lead to hepatic insulin resistance; these effects seem specific to fructose, and potential mechanisms may involve both stimulation of hepatic de novo lipogenesis and decreased lipid oxidation. Moreover, the results suggest that women seem protected against such deleterious effects, while OffT2D displayed exacerbated response. Résumé : Le fructose est un sucre simple, dont la consommation a augmenté durant les dernières décennies. Dans les modèles animaux, un régime riche en fructose (RRFru) peut induire plusieurs composantes du syndrome métabolique. Le but de cette thèse était d'étudier les effets d'un régime riche en fructose sur la sensibilité à l'insuline et la déposition de lipides ectopiques chez l'humain, et si ces effets variaient selon le genre ou le background génétique. La première étude avait pour but d'évaluer l'effet d'un RRFru d'une durée de 4 semaines sur la sensibilité à l'insuline et le métabolisme des lipides chez des hommes sains. La sensibilité à l'insuline, les lipides intrahépatiques (IHCL) et intramusculaires (IMCL) ont été mesurés avant, et après 1 et 4 semaines du RRFru (1.5 g fructose/kg/jour). La sensibilité à l'insuline a été déterminée par un clamp hyperinsulinémique euglycémique, et les IHCL/IMCL par spectroscopie à résonnance magnétique. Le fructose a augmenté les concentrations plasmatiques à jeun des VLDL- triglycérides (TG) (+72%) et de glucose (+6%), sans induire de changement au niveau de la sensibilité à l'insuline, IHCL ou IMCL. Dans la deuxième étude, des biopsies de muscle squelettique ont été prélevées chez cinq de ces volontaires avant et après les 4 semaines de RRFru. Les concentrations de mRNA de 18 gènes impliqués dans le métabolisme des lipides et des hydrates de carbone ont été mesurées par RT-PCR quantitative. Le RRFru a augmenté l'expression de gènes impliqués dans la synthèse de lipides, et diminué celles de gènes impliqués dans la sensibilité à l'insuline et l'oxydation de lipides. Ces changements pourraient constituer des altérations précoces de la sensibilité à l'insuline et du métabolisme lipidique en réponse au fructose. La troisième étude avait pour but de définir si les réponses au RRFru étaient semblables entre les hommes et les femmes. Pour ceci, des doses plus élevées de fructose ont été administrées à 8 jeunes hommes et 8 jeunes femmes durant 6 jours. Le RRFru a augmenté les TG chez les hommes (+71 %), et de manière nettement plus modeste chez les femmes (+16%). Les hommes ont développé une résistance hépatique à l'insuline, ainsi qu'une augmentation des concentrations d'alanine aminotransférase (+38%), mais aucun de ces effets n'a été observé chez les femmes. Cette étude suggère qu'à court terme, un RRFru mène à une hypertriglycéridémie et résistance hépatique à l'insuline chez l'homme, tandis que les femmes semblent en être protégées. Finalement, la 4ème étude a investigué si des personnes apparentées à des patients diabétiques de type 2 (AppDT2), qui constituent un groupe d'individus à risque de développer des maladies métaboliques en raison de leur background génétique, avaient des réponses plus marquées au RRFru. Huit hommes sains (Ctrl) et 16 AppDT2 on reçu dans un ordre randomisé un RRFru et une diète isocalorique durant 6 jours. Dans les deux groupes, le RRFru a augmenté significativement les IHCL (Ctrl: +76%; AppDT2: +79%) et les VLDL-TG plasmatiques à jeun (Ctrl: +51%; AppDT2: +110%). En valeurs absolues, ces deux augmentations étaient plus importantes dans le groupe des AppDT2, suggérant que ces individus sont plus à risque de développer des problèmes métaboliques suite à un apport de fructose. Afin de définir les effets spécifiques du fructose, quatre des huit sujets Ctrl ont été soumis à un régime riche en glucose. Après le régime riche en glucose, les concentrations de TG et d'IHCL étaient semblables aux valeurs obtenues après une diète isocalorique, contrairement aux nombreux effets observés après le RRFru. En conclusion, ces différentes études ont démontré que chez l'humain, le fructose peut rapidement induire une augmentation des VLDL-TG à jeun, des IHCL et une résistance hépatique à l'insuline ; ces effets semblent être spécifiques au fructose. De plus, les différents résultats obtenus montrent que les femmes développent des effets moindres en réponse au fructose, contrairement aux AppDT2, chez qui les effets du fructose semblent plus marqués. Résumé grand public : Le fructose est un sucre simple, présent naturellement et en faibles quantités dans les fruits, mais également constituant du sucrose - appelé aussi sucre de table. Depuis les années 1970, la consommation de fructose a augmenté dans les pays industrialisés et émergents, principalement par le biais d'une hausse de consommation de boissons sucrées de type soda. Dans des modèles animaux tels que les rongeurs, un régime riche en fructose mène au développement de plusieurs facteurs de risques étroitement liés aux maladies cardiovasculaires, à l'obésité et au diabète de type 2; ceux-ci sont caractérisés par une augmentation des concentrations de glucose et de lipides sanguins, ainsi qu'une accumulation de lipides dits « ectopiques », à savoir dans le foie et les muscles. Le but de cette thèse était de définir les effets d'un régime riche en fructose chez l'être humain. De plus, nous nous sommes intéressés à savoir si ces effets étaient semblables entre différents groupes d'individus, à savoir des personnes de sexe masculin / féminin, ou des personnes dont au moins un des parents est diabétique de type 2. Pour ceci, différents groupes de volontaires (hommes, femmes, avec histoire familiale de diabète de type 2) âgés de 18-30 ans se sont soumis à une alimentation enrichie en fructose, d'une durée allant de 6 à 28 jours, suivant l'étude à laquelle ils participaient. La quantité de fructose consommée en plus de l'alimentation normale durant ces périodes équivalait au contenu en fructose de 2-4 litres de boissons sucrées par jour. Des prises de sang ont été effectuées au terme de chacun de ces différents régimes, ainsi que des mesures de sensibilité à l'insuline et de concentrations de lipides dans le foie et le muscle par résonnance magnétique nucléaire, en collaboration avec l'Hôpital de l'Ile de Berne. Les résultats montrent qu'après 6 jours de régime riche en fructose, les volontaires sains de sexe masculin ont presque doublé leurs concentrations de lipides sanguins et hépatiques. De plus, le foie de ces volontaires réagissait moins bien à l'insuline, ce qui pourrait mener à long terme à des maladies métaboliques comme le diabète de type 2. Un des mécanismes postulés est que le fructose pourrait stimuler la formation de lipides dans le foie, contribuant ainsi à un dysfonctionnement de cet organe. De manière surprenante, des femmes d'âge et d'IMC (Indice de Masse Corporelle) comparables aux hommes étudiés n'ont pas développé ces différents effets en réponse au régime riche en fructose. Il semblerait donc qu'elles possèdent certaines propriétés pouvant les «protéger », du moins à court terme, des problèmes métaboliques induits par le fructose. De tels mécanismes sont pour l'heure inconnus, mais il est possible que des différences hormonales, ou de répartition de la masse graisseuse dans le corps, puissent jouer un rôle. Enfin, nous avons également démontré que chez certaines personnes ayant au moins un parent (père ou mère) diabétique de type 2, les augmentations de lipides sanguins et hépatiques induits par le fructose étaient plus marquées que chez des volontaires sans parents diabétiques. Ceci est néanmoins à tempérer par le fait que nous avons observé une grande hétérogénéité des réponses parmi ces individus, découlant certainement d'interactions complexes entre différents facteurs tels que la génétique, le mode de vie, l'alimentation et l'activité physique. Ces différents résultats donnent lieu à une meilleure compréhension du rôle de facteurs alimentaires dans le développement de problèmes métaboliques tels que le diabète de type 2. Ils vont également permettre de tester différentes approches thérapeutiques. Bien qu'ayant été obtenus avec des doses de fructose importantes, ces études soulignent l'effet potentiellement dangereux pour la santé d'une alimentation riche en sucres.

Anatomic and functional outcome after 23-gauge sutureless transconjunctival vitrectomy, peeling and intravitreal triamcinolone for idiopathic macular epiretinal membranes

Purpose:To report the functional, anatomic outcome and safety profile of 23-gauge pars plana vitrectomy (PPV) combined with peeling and intravitreal injection of triamcinolone acetonide (TA) in eyes with idiopathic epiretinal membranes (ERM). Methods:Retrospective, nonrandomized study of consecutive patients who underwent 23-gauge transconjunctival sutureless PPV with subsequent membrane peeling and intravitreal TA injection for an idiopathic ERM. All patients were operated between February 2007 and February 2008 at the Jules Gonin University Eye Hospital. The minimum follow-up was 6-months. Results:Thirty-nine eyes of 39 patients were included. The mean follow-up was 7 months (SD: 2.2, range: 6-15 months). Twenty-two (56%) eyes were pseudophakic and 17 (44%) were phakic at the time of surgery. Mean preoperative intraocular pressure (IOP) was 14 mmHg (SD: 3.5). At the final follow-up mean IOP was 14.5 (SD: 2.7) that did not differ significantly from the IOP at baseline (P: 0.14- 2-tailed t test). Five patients (13%) needed temporary topical anti-glaucoma treatment.Mean preoperative BCVA was 0.28 decimal equivalent (logMAR 0.54, SD: 0.2, range: 1.0 - 0.2). and improved significantly (P <0.0001, 2-tailed t test) to a mean of 0.6 decimal equivalent (logMAR 0.22, SD: 0.16, range: 0.6 - 0) at the final follow-up. The visual acuity improved by a mean of 3.2 lines (SD 2.1, range 0- 8). Twenty-nine patients (74%) demonstrated a gain of 3 or more lines.Mean central macular thickness (CMT) was 456 µm (SD: 77) at the baseline that reduced significantly (P <0.0001, 2-tailed t test) at the final follow-up to 327µm (SD: 79). Average CMT reduction was 131µm (SD: 77, range: 36- 380 µm). A subgroup analysis of 15 selected cases that had CMT measurement 1 week after surgery demonstrated that 84% of the total final reduction in CMT occurred during the first week. Conclusions:23-gauge sutureless transconjunctival vitrectomy with the concomitant administration of intravitreal TA is a safe and effective technique for the treatment of idiopathic ERM and may speed up anatomical recovery.

Association between pre-diagnostic circulating vitamin D concentration and risk of colorectal cancer in European populations:a nested case-control study

Objective. To examine the association between pre-diagnostic circulating vitamin D concentration, dietary intake of vitamin D and calcium, and the risk of colorectal cancer in European populations. Design Nested case-control study. Setting. The study was conducted within the EPIC study, a cohort of more than 520 000 participants from 10 western European countries. Participants: 1248 cases of incident colorectal cancer, which developed after enrolment into the cohort, were matched to 1248 controls. Main outcome measures. Circulating vitamin D concentration (25-hydroxy-vitamin-D, 25-(OH)D) was measured by enzyme immunoassay. Dietary and lifestyle data were obtained from questionnaires. Incidence rate ratios and 95% confidence intervals for the risk of colorectal cancer by 25-(OH)D concentration and levels of dietary calcium and vitamin D intake were estimated from multivariate conditional logistic regression models, with adjustment for potential dietary and other confounders. Results. 25-(OH)D concentration showed a strong inverse linear dose-response association with risk of colorectal cancer (P for trend <0.001). Compared with a pre-defined mid-level concentration of 25-(OH)D (50.0-75.0 nmol/l), lower levels were associated with higher colorectal cancer risk (<25.0 nmol/l: incidence rate ratio 1.32 (95% confidence interval 0.87 to 2.01); 25.0-49.9 nmol/l: 1.28 (1.05 to 1.56), and higher concentrations associated with lower risk (75.0-99.9 nmol/l: 0.88 (0.68 to 1.13); ≥100.0 nmol/l: 0.77 (0.56 to 1.06)). In analyses by quintile of 25-(OH)D concentration, patients in the highest quintile had a 40% lower risk of colorectal cancer than did those in the lowest quintile (P<0.001). Subgroup analyses showed a strong association for colon but not rectal cancer (P for heterogeneity=0.048). Greater dietary intake of calcium was associated with a lower colorectal cancer risk. Dietary vitamin D was not associated with disease risk. Findings did not vary by sex and were not altered by corrections for season or month of blood donation. Conclusions The results of this large observational study indicate a strong inverse association between levels of pre-diagnostic 25-(OH)D concentration and risk of colorectal cancer in western European populations. Further randomised trials are needed to assess whether increases in circulating 25-(OH)D concentration can effectively decrease the risk of colorectal cancer.

Echovirus 30 associated with cases of aseptic meningitis in state of Pará, Northern Brazil

Investigation of the aetiology of viral meningitis in Brazil is most often restricted to cases that occur in the Southern and Southeastern Regions; therefore, the purpose of this study is to describe the viral meningitis cases that occurred in state of Pará, Northern Brazil, from January 2005-December 2006. The detection of enterovirus (EV) in cerebrospinal fluid was performed using cell culture techniques, RT-PCR, nested PCR and nucleotide sequencing. The ages of the 91 patients ranged from < one year old to > 60 years old (median age 15.90 years). Fever (87.1%), headache (77.0%), vomiting (61.5%) and stiffness (61.5%) were the most frequent symptoms. Of 91 samples analyzed, 18 (19.8%) were positive for EV. Twelve were detected only by RT- PCR followed by nested PCR, whereas six were found by both cell culture and RT-PCR. From the last group, five were sequenced and classified as echovirus 30 (Echo 30). Phylogenetic analyses revealed that Echo 30 detected in Northern Brazil clustered within a unique group with a bootstrap value of 100% and could constitute a new subgroup (4c) according to the phylogenetic tree described by Oberste et al. (1999). This study described the first molecular characterization of Echo 30 in Brazil and this will certainly contribute to future molecular analyses involving strains detected in other regions of Brazil.

CD209 in inflammatory bowel disease: a case-control study in the Spanish population

BACKGROUND The etiology of Ulcerative Colitis (UC) and Crohn's Disease (CD), considered together as Inflammatory Bowel Diseases (IBD), involves environmental and genetic factors. Although some genes are already known, the genetics underlying these diseases is complex and new candidates are continuously emerging. The CD209 gene is located in a region linked previously to IBD and a CD209 functional polymorphism (rs4804803) has been associated to other inflammatory conditions. Our aim was to study the potential involvement of this CD209 variant in IBD susceptibility. METHODS We performed a case-control study with 515 CD patients, 497 UC patients and 731 healthy controls, all of them white Spaniards. Samples were typed for the CD209 single nucleotide polymorphism (SNP) rs4804803 by TaqMan technology. Frequency comparisons were performed using chi2 tests. RESULTS No association between CD209 and UC or CD was observed initially. However, stratification of UC patients by HLA-DR3 status, a strong protective allele, showed that carriage of the CD209_G allele could increase susceptibility in the subgroup of HLA-DR3-positive individuals (p = 0.03 OR = 1.77 95% CI 1.04-3.02, vs. controls). CONCLUSION A functional variant in the CD209 gene, rs4804803, does not seem to be influencing Crohn's disease susceptibility. However, it could be involved in the etiology or pathology of Ulcerative Colitis in HLA-DR3-positive individuals but further studies are necessary.

An iterative Multiscale Finite-Volume algorithm converging to the exact solution

The multiscale finite volume (MsFV) method has been developed to efficiently solve large heterogeneous problems (elliptic or parabolic); it is usually employed for pressure equations and delivers conservative flux fields to be used in transport problems. The method essentially relies on the hypothesis that the (fine-scale) problem can be reasonably described by a set of local solutions coupled by a conservative global (coarse-scale) problem. In most cases, the boundary conditions assigned for the local problems are satisfactory and the approximate conservative fluxes provided by the method are accurate. In numerically challenging cases, however, a more accurate localization is required to obtain a good approximation of the fine-scale solution. In this paper we develop a procedure to iteratively improve the boundary conditions of the local problems. The algorithm relies on the data structure of the MsFV method and employs a Krylov-subspace projection method to obtain an unconditionally stable scheme and accelerate convergence. Two variants are considered: in the first, only the MsFV operator is used; in the second, the MsFV operator is combined in a two-step method with an operator derived from the problem solved to construct the conservative flux field. The resulting iterative MsFV algorithms allow arbitrary reduction of the solution error without compromising the construction of a conservative flux field, which is guaranteed at any iteration. Since it converges to the exact solution, the method can be regarded as a linear solver. In this context, the schemes proposed here can be viewed as preconditioned versions of the Generalized Minimal Residual method (GMRES), with a very peculiar characteristic that the residual on the coarse grid is zero at any iteration (thus conservative fluxes can be obtained).

Prevalence and genotyping of hepatitis C virus in blood donors in the state of Pará, Northern Brazil

Given the scarcity of epidemiological information on hepatitis C virus (HCV) infection in Northern Brazil, we determined the prevalence and genotypic frequency in blood donors in the state of Pará (PA). Blood samples from all of the blood donors at the Fundação HEMOPA (blood bank of PA) from 2004-2006 were screened for the presence of antibodies to anti-HCV and samples seroreactive to anti-HCV were further tested for HCV RNA using real-time PCR. In total, 116 HCV-RNA samples were genotyped, based on maximum likelihood phylogenetic analyses, using BioEdit, Modelgenerator, PHYML and FigTree software. The population consisted of 242,726 volunteers who donated blood from 2004-2006; the most common subgroup was males between the ages of 18-29 years old (37.30%). Within the whole group, 1,112 blood donors (0.46%) had indeterminate or positive serology; among these, 28.78% were males whose ages ranged from 18-29 years. A diagnosis of chronic HCV infection was confirmed for 304 donors (60.20% males; 66.45% were 30-49 years old), resulting in a prevalence of HCV RNA in 0.13% of the samples (304 of 242,726). HCV genotyping revealed a high frequency of genotype 1 (108/116) followed by genotype 3 (8/116). This study found HCV infection to be relatively infrequent in PA; genotype 1 was most commonly isolated. This information can help guide prevention and control policies aimed at efficient diagnosis and control measures.

Genome-wide analysis of SREBP1 activity around the clock reveals its combined dependency on nutrient and circadian signals.

In mammals, the circadian clock allows them to anticipate and adapt physiology around the 24 hours. Conversely, metabolism and food consumption regulate the internal clock, pointing the existence of an intricate relationship between nutrient state and circadian homeostasis that is far from being understood. The Sterol Regulatory Element Binding Protein 1 (SREBP1) is a key regulator of lipid homeostasis. Hepatic SREBP1 function is influenced by the nutrient-response cycle, but also by the circadian machinery. To systematically understand how the interplay of circadian clock and nutrient-driven rhythm regulates SREBP1 activity, we evaluated the genome-wide binding of SREBP1 to its targets throughout the day in C57BL/6 mice. The recruitment of SREBP1 to the DNA showed a highly circadian behaviour, with a maximum during the fed status. However, the temporal expression of SREBP1 targets was not always synchronized with its binding pattern. In particular, different expression phases were observed for SREBP1 target genes depending on their function, suggesting the involvement of other transcription factors in their regulation. Binding sites for Hepatocyte Nuclear Factor 4 (HNF4) were specifically enriched in the close proximity of SREBP1 peaks of genes, whose expression was shifted by about 8 hours with respect to SREBP1 binding. Thus, the cross-talk between hepatic HNF4 and SREBP1 may underlie the expression timing of this subgroup of SREBP1 targets. Interestingly, the proper temporal expression profile of these genes was dramatically changed in Bmal1-/- mice upon time-restricted feeding, for which a rhythmic, but slightly delayed, binding of SREBP1 was maintained. Collectively, our results show that besides the nutrient-driven regulation of SREBP1 nuclear translocation, a second layer of modulation of SREBP1 transcriptional activity, strongly dependent from the circadian clock, exists. This system allows us to fine tune the expression timing of SREBP1 target genes, thus helping to temporally separate the different physiological processes in which these genes are involved.

Guidelines for specialized nutritional and metabolic support in the critically-ill patient. Update. Consensus SEMICYUC-SENPE: Obese patient

As a response to metabolic stress, obese critically-ill patients have the same risk of nutritional deficiency as the non-obese and can develop protein-energy malnutrition with accelerated loss of muscle mass. The primary aim of nutritional support in these patients should be to minimize loss of lean mass and accurately evaluate energy expenditure. However, routinely used formulae can overestimate calorie requirements if the patient's actual weight is used. Consequently, the use of adjusted or ideal weight is recommended with these formulae, although indirect calorimetry is the method of choice. Controversy surrounds the question of whether a strict nutritional support criterion, adjusted to the patient's requirements, should be applied or whether a certain degree of hyponutrition should be allowed. Current evidence suggested that hypocaloric nutrition can improve results, partly due to a lower rate of infectious complications and better control of hyperglycemia. Therefore, hypocaloric and hyperproteic nutrition, whether enteral or parenteral, should be standard practice in the nutritional support of critically-ill obese patients when not contraindicated. Widely accepted recommendations consist of no more than 60-70% of requirements or administration of 11-14 kcal/kg current body weight/day or 22-25 kcal/kg ideal weight/day, with 2-2.5 g/kg ideal weight/day of proteins. In a broad sense, hypocaloric-hyperprotein regimens can be considered specific to obese critically-ill patients, although the complications related to comorbidities in these patients may require other therapeutic possibilities to be considered, with specific nutrients for hyperglycemia, acute respiratory distress syndrome (ARDS) and sepsis. However, there are no prospective randomized trials with this type of nutrition in this specific population subgroup and the available data are drawn from the general population of critically-ill patients. Consequently, caution should be exercised when interpreting these data.

Guidelines for specialized nutritional and metabolic support in the critically-ill patient. Update. Consensus SEMICYUC-SENPE: Septic patient

Nutritional metabolic management, together with other treatment and support measures used, is one of the mainstays of the treatment of septic patients. Nutritional support should be started early, after initial life support measures, to avoid the consequences of malnutrition, to provide adequate nutritional intake and to prevent the development of secondary complications such as superinfection or multiorgan failure. As in other critically-ill patients, when the enteral route cannot be used to ensure calorie-protein requirements, the association of parenteral nutrition has been shown to be safe in this subgroup of patients. Studies evaluating the effect of specific pharmaconutrients in septic patients are scarce and are insufficient to allow recommendations to be made. To date, enteral diets with a mixture of substrates with distinct pharmaconutrient properties do not seem to be superior to standard diets in altering the course of sepsis, although equally there is no evidence that these diets are harmful. There is insufficient evidence to recommend the use of glutamine in septic patients receiving parenteral nutrition. However, given the good results and absence of glutamine-related adverse effects in the various studies performed in the general population of critically-ill patients, these patients could benefit from the use of this substance. Routine use of omega-3 fatty acids cannot be recommended until further evidence has been gathered, although the use of lipid emulsions with a high omega-6 fatty acid content should be avoided. Septic patients should receive an adequate supply of essential trace elements and vitamins. Further studies are required before the use of high-dose selenium can be recommended.

Additive effects of LPL, APOA5 and APOE variant combinations on triglyceride levels and hypertriglyceridemia: results of the ICARIA genetic sub-study.

BACKGROUND Hypertriglyceridemia (HTG) is a well-established independent risk factor for cardiovascular disease and the influence of several genetic variants in genes related with triglyceride (TG) metabolism has been described, including LPL, APOA5 and APOE. The combined analysis of these polymorphisms could produce clinically meaningful complementary information. METHODS A subgroup of the ICARIA study comprising 1825 Spanish subjects (80% men, mean age 36 years) was genotyped for the LPL-HindIII (rs320), S447X (rs328), D9N (rs1801177) and N291S (rs268) polymorphisms, the APOA5-S19W (rs3135506) and -1131T/C (rs662799) variants, and the APOE polymorphism (rs429358; rs7412) using PCR and restriction analysis and TaqMan assays. We used regression analyses to examine their combined effects on TG levels (with the log-transformed variable) and the association of variant combinations with TG levels and hypertriglyceridemia (TG > or = 1.69 mmol/L), including the covariates: gender, age, waist circumference, blood glucose, blood pressure, smoking and alcohol consumption. RESULTS We found a significant lowering effect of the LPL-HindIII and S447X polymorphisms (p < 0.0001). In addition, the D9N, N291S, S19W and -1131T/C variants and the APOE-epsilon4 allele were significantly associated with an independent additive TG-raising effect (p < 0.05, p < 0.01, p < 0.001, p < 0.0001 and p < 0.001, respectively). Grouping individuals according to the presence of TG-lowering or TG-raising polymorphisms showed significant differences in TG levels (p < 0.0001), with the lowest levels exhibited by carriers of two lowering variants (10.2% reduction in TG geometric mean with respect to individuals who were homozygous for the frequent alleles of all the variants), and the highest levels in carriers of raising combinations (25.1% mean TG increase). Thus, carrying two lowering variants was protective against HTG (OR = 0.62; 95% CI, 0.39-0.98; p = 0.042) and having one single raising polymorphism (OR = 1.20; 95% CI, 1.39-2.87; p < 0.001) or more (2 or 3 raising variants; OR = 2.90; 95% CI, 1.56-5.41; p < 0.001) were associated with HTG. CONCLUSION Our results showed a significant independent additive effect on TG levels of the LPL polymorphisms HindIII, S447X, D9N and N291S; the S19W and -1131T/C variants of APOA5, and the epsilon4 allele of APOE in our study population. Moreover, some of the variant combinations studied were significantly associated with the absence or the presence of hypertriglyceridemia.

Influence of environmental conditions on the distribution of Central Asian green toads with three ploidy levels.

We studied the distribution of Palearctic green toads (Bufo viridis subgroup), an anuran species group with three ploidy levels, inhabiting the Central Asian Amudarya River drainage. Various approaches (one-way, multivariate, components variance analyses and maximum entropy modelling) were used to estimate the effect of altitude, precipitation, temperature and land vegetation covers on the distribution of toads. It is usually assumed that polyploid species occur in regions with harsher climatic conditions (higher latitudes, elevations, etc.), but for the green toads complex, we revealed a more intricate situation. The diploid species (Bufo shaartusiensis and Bufo turanensis) inhabit the arid lowlands (from 44 to 789 m a.s.l.), while tetraploid Bufo pewzowi were recorded in mountainous regions (340-3492 m a.s.l.) with usually lower temperatures and higher precipitation rates than in the region inhabited by diploid species. The triploid species Bufo baturae was found in the Pamirs (Tajikistan) at the highest altitudes (2503-3859 m a.s.l.) under the harshest climatic conditions.

Long-term risk of incident type 2 diabetes and measures of overall and regional obesity: the EPIC-InterAct case-cohort study

BACKGROUND Waist circumference (WC) is a simple and reliable measure of fat distribution that may add to the prediction of type 2 diabetes (T2D), but previous studies have been too small to reliably quantify the relative and absolute risk of future diabetes by WC at different levels of body mass index (BMI). METHODS AND FINDINGS The prospective InterAct case-cohort study was conducted in 26 centres in eight European countries and consists of 12,403 incident T2D cases and a stratified subcohort of 16,154 individuals from a total cohort of 340,234 participants with 3.99 million person-years of follow-up. We used Prentice-weighted Cox regression and random effects meta-analysis methods to estimate hazard ratios for T2D. Kaplan-Meier estimates of the cumulative incidence of T2D were calculated. BMI and WC were each independently associated with T2D, with WC being a stronger risk factor in women than in men. Risk increased across groups defined by BMI and WC; compared to low normal weight individuals (BMI 18.5-22.4 kg/m(2)) with a low WC (<94/80 cm in men/women), the hazard ratio of T2D was 22.0 (95% confidence interval 14.3; 33.8) in men and 31.8 (25.2; 40.2) in women with grade 2 obesity (BMI≥35 kg/m(2)) and a high WC (>102/88 cm). Among the large group of overweight individuals, WC measurement was highly informative and facilitated the identification of a subgroup of overweight people with high WC whose 10-y T2D cumulative incidence (men, 70 per 1,000 person-years; women, 44 per 1,000 person-years) was comparable to that of the obese group (50-103 per 1,000 person-years in men and 28-74 per 1,000 person-years in women). CONCLUSIONS WC is independently and strongly associated with T2D, particularly in women, and should be more widely measured for risk stratification. If targeted measurement is necessary for reasons of resource scarcity, measuring WC in overweight individuals may be an effective strategy, since it identifies a high-risk subgroup of individuals who could benefit from individualised preventive action.