Genetic variants of HvCbf14 are statistically associated with frost tolerance in a European germplasm collection of Hordeum vulgare

Two quantitative trait loci (Fr-H1 and Fr-H2) for frost tolerance (FT) have been discovered on the long arm of chromosome 5H in barley. Two tightly linked groups of CBF genes, known to play a key role in the FT regulatory network in A. thaliana, have been found to co-segregate with Fr-H2. Here, we investigate the allelic variations of four barley CBF genes (HvCbf3, HvCbf6, HvCbf9 and HvCbf14) in a panel of European cultivars, landraces and H. spontaneum accessions. In the cultivars a reduction of nucleotide and haplotype diversities in CBFs compared with the landraces and the wild ancestor H. spontaneum, was evident. In particular, in cultivars the loss of HvCbf9 genetic variants was higher compared to other sequences. In order to verify if the pattern of CBF genetic variants correlated with the level of FT, an association procedure was adopted. The pairwise analysis of linkage disequilibrium (LD) among the genetic variants in four CBF genes was computed to evaluate the resolution of the association procedure. The pairwise plotting revealed a low level of LD in cultivated varieties, despite the tight physical linkage of CBF genes analysed. A structured association procedure based on a general liner model was implemented, including the variants in CBFs, of Vrn-H1, and of two reference genes not involved in FT (α-Amy1 and Gapdh) and considering the phenotypic data for FT. Association analysis recovered two nucleotide variants of HvCbf14 and one nucleotide variant of Vrn-H1 as statistically associated to FT

Genome-Wide Analysis of Glucocorticoid Receptor Binding Regions in Adipocytes Reveal Gene Network Involved in Triglyceride Homeostasis

Glucocorticoids play important roles in the regulation of distinct aspects of adipocyte biology. Excess glucocorticoids in adipocytes are associated with metabolic disorders, including central obesity, insulin resistance and dyslipidemia. To understand the mechanisms underlying the glucocorticoid action in adipocytes, we used chromatin immunoprecipitation sequencing to isolate genome-wide glucocorticoid receptor (GR) binding regions (GBRs) in 3T3-L1 adipocytes. Furthermore, gene expression analyses were used to identify genes that were regulated by glucocorticoids. Overall, 274 glucocorticoid-regulated genes contain or locate nearby GBR. We found that many GBRs were located in or nearby genes involved in triglyceride (TG) synthesis (Scd-1, 2, 3, GPAT3, GPAT4, Agpat2, Lpin1), lipolysis (Lipe, Mgll), lipid transport (Cd36, Lrp-1, Vldlr, Slc27a2) and storage (S3-12). Gene expression analysis showed that except for Scd-3, the other 13 genes were induced in mouse inguinal fat upon 4-day glucocorticoid treatment. Reporter gene assays showed that except Agpat2, the other 12 glucocorticoid-regulated genes contain at least one GBR that can mediate hormone response. In agreement with the fact that glucocorticoids activated genes in both TG biosynthetic and lipolytic pathways, we confirmed that 4-day glucocorticoid treatment increased TG synthesis and lipolysis concomitantly in inguinal fat. Notably, we found that 9 of these 12 genes were induced in transgenic mice that have constant elevated plasma glucocorticoid levels. These results suggested that a similar mechanism was used to regulate TG homeostasis during chronic glucocorticoid treatment. In summary, our studies have identified molecular components in a glucocorticoid-controlled gene network involved in the regulation of TG homeostasis in adipocytes. Understanding the regulation of this gene network should provide important insight for future therapeutic developments for metabolic diseases

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Raised serum TSH in morbid-obese and non-obese patients: effect on the circulating lipid profile

Morbid obesity is associated with a high rate of raised serum TSH associated with normal free thyroid hormones. The body repercussions of this thyroid abnormality, suggesting subclinical hypothyroidism, are still debated. In particular, it is unclear whether the raised serum TSH of obesity results in changes of circulating lipids typically observed in hypothyroidism. Aim of this study was to evaluate the impact of a raised serum TSH on the lipid profile in morbid-obese and non-obese patients. Serum TSH, FT4, FT3, Tg-Ab, TPO-Ab and lipids were measured in 55 morbid-obese (BMI > 40 kg/m(2)) and 55 non-obese (BMI < 30 kg/m(2)) patients with a raised serum TSH. Despite similar serum levels of TSH, FT4 and FT3, morbid-obese patients displayed significantly lower mean levels of total cholesterol (200.8 ± 35.6 vs. 226.9 ± 41.4 mg/dl, p < 0.001) and a significantly lower prevalence of hypercholesterolemia (50.9 vs. 72.7 %, p < 0.01) when compared with non-obese patients. Morbid-obese patients also had lower mean serum HDL cholesterol and higher serum triglycerides. The impact of a raised serum TSH on the lipid profile differs in morbid-obese compared to non-obese patients, suggesting that obese patients might not be truly hypothyroid. Measuring total cholesterol could be a helpful tool for deciding whether a morbid-obese patient with a raised serum TSH should be given levothyroxine treatment

Raised serum TSH in morbid-obese and non-obese patients: effect on the circulating lipid profile.

Morbid obesity is associated with a high rate of raised serum TSH associated with normal free thyroid hormones. The body repercussions of this thyroid abnormality, suggesting subclinical hypothyroidism, are still debated. In particular, it is unclear whether the raised serum TSH of obesity results in changes of circulating lipids typically observed in hypothyroidism. Aim of this study was to evaluate the impact of a raised serum TSH on the lipid profile in morbid-obese and non-obese patients. Serum TSH, FT4, FT3, Tg-Ab, TPO-Ab and lipids were measured in 55 morbid-obese (BMI > 40 kg/m2) and 55 non-obese (BMI < 30 kg/m2) patients with a raised serum TSH. Despite similar serum levels of TSH, FT4 and FT3, morbid-obese patients displayed significantly lower mean levels of total cholesterol (200.8 ± 35.6 vs. 226.9 ± 41.4 mg/dl, p < 0.001) and a significantly lower prevalence of hypercholesterolemia (50.9 vs. 72.7 %, p < 0.01) when compared with non-obese patients. Morbid-obese patients also had lower mean serum HDL cholesterol and higher serum triglycerides. The impact of a raised serum TSH on the lipid profile differs in morbid-obese compared to non-obese patients, suggesting that obese patients might not be truly hypothyroid. Measuring total cholesterol could be a helpful tool for deciding whether a morbid-obese patient with a raised serum TSH should be given levothyroxine treatment