Inhibition of SIRT1 Impairs the Accumulation and Transcriptional Activity of HIF-1α Protein under Hypoxic Conditions

Sirtuins and hypoxia-inducible transcription factors (HIF) have well-established roles in regulating cellular responses to metabolic and oxidative stress. Recent reports have linked these two protein families by demonstrating that sirtuins can regulate the activity of HIF-1 and HIF-2. Here we investigated the role of SIRT1, a NAD+-dependent deacetylase, in the regulation of HIF-1 activity in hypoxic conditions. Our results show that in hepatocellular carcinoma (HCC) cell lines, hypoxia did not alter SIRT1 mRNA or protein expression, whereas it predictably led to the accumulation of HIF-1α and the up-regulation of its target genes. In hypoxic models in vitro and in in vivo models of systemic hypoxia and xenograft tumor growth, knockdown of SIRT1 protein with shRNA or inhibition of its activity with small molecule inhibitors impaired the accumulation of HIF-1α protein and the transcriptional increase of its target genes. In addition, endogenous SIRT1 and HIF-1α proteins co-immunoprecipitated and loss of SIRT1 activity led to a hyperacetylation of HIF-1α. Taken together, our data suggest that HIF-1α and SIRT1 proteins interact in HCC cells and that HIF-1α is a target of SIRT1 deacetylase activity. Moreover, SIRT1 is necessary for HIF-1α protein accumulation and activation of HIF-1 target genes under hypoxic conditions

Mapping variants in thyroid hormone transporter MCT8 to disease severity by genomic, phenotypic, functional, structural and deep learning integration

Predicting and quantifying phenotypic consequences of genetic variants in rare disorders is a major challenge, particularly pertinent for 'actionable' genes such as thyroid hormone transporter MCT8 (encoded by the X-linked SLC16A2 gene), where loss-of-function (LoF) variants cause a rare neurodevelopmental and (treatable) metabolic disorder in males. The combination of deep phenotyping data with functional and computational tests and with outcomes in population cohorts, enabled us to: (i) identify the genetic aetiology of divergent clinical phenotypes of MCT8 deficiency with genotype-phenotype relationships present across survival and 24 out of 32 disease features; (ii) demonstrate a mild phenocopy in ~400,000 individuals with common genetic variants in MCT8; (iii) assess therapeutic effectiveness, which did not differ among LoF-categories; (iv) advance structural insights in normal and mutated MCT8 by delineating seven critical functional domains; (v) create a pathogenicity-severity MCT8 variant classifier that accurately predicted pathogenicity (AUC:0.91) and severity (AUC:0.86) for 8151 variants. Our information-dense mapping provides a generalizable approach to advance multiple dimensions of rare genetic disorders

Mapping variants in thyroid hormone transporter MCT8 to disease severity by genomic, phenotypic, functional, structural and deep learning integration

Predicting and quantifying phenotypic consequences of genetic variants in rare disorders is a major challenge, particularly pertinent for ‘actionable’ genes such as thyroid hormone transporter MCT8 (encoded by the X-linked SLC16A2 gene), where loss-of-function (LoF) variants cause a rare neurodevelopmental and (treatable) metabolic disorder in males. The combination of deep phenotyping data with functional and computational tests and with outcomes in population cohorts, enabled us to: (i) identify the genetic aetiology of divergent clinical phenotypes of MCT8 deficiency with genotype-phenotype relationships present across survival and 24 out of 32 disease features; (ii) demonstrate a mild phenocopy in ~400,000 individuals with common genetic variants in MCT8; (iii) assess therapeutic effectiveness, which did not differ among LoF-categories; (iv) advance structural insights in normal and mutated MCT8 by delineating seven critical functional domains; (v) create a pathogenicity-severity MCT8 variant classifier that accurately predicted pathogenicity (AUC:0.91) and severity (AUC:0.86) for 8151 variants. Our information-dense mapping provides a generalizable approach to advance multiple dimensions of rare genetic disorders

Gaps in smiles and services: a cross-sectional study of dental caries in refugee-background children

BACKGROUND: Refugees are reported to experience high rates of dental disease, although there are limited data on refugee children. The aim of this study was to report on oral health in refugee-background children in Australia, and to assess their follow-up at dental services. METHODS: Cross-sectional study of opportunistic oral health screening and subsequent dental service use in refugee-background children attending a refugee health clinic in Victoria, Australia, between November 2006-November 2010. RESULTS: 350 patients (0-18 years, mean age 8 years 7 months) had oral health screening; 241 (68.9%) were born overseas, (176 Africa, 65 other countries) and 109 (31.1%) were born in Australia to African-background families. Parents were concerned about oral health in 65/341 (19.1%) children, with specific concern about caries in only 9/341 (2.6%). On assessment, 155/336 (46.1%) had visible caries and 178/345 (51.6%) had caries experience (dmft/DMFT > 0). Where parents were concerned about caries, they were likely to be present (positive predictive value = 100%), however absence of parent concern about caries was not reassuring (negative predictive value = 56.1%).Compared to Australian-born children of African background; African-born children were more likely to be referred for further dental care (adjusted PR 1.33, 95% CI [1.02-1.73]), although there was no statistically significant difference in caries prevalence. African-born children were less likely to have caries compared to other overseas-born children (adjusted PR 0.73, 95% CI [0.58 - 0.93]). Overall 187/344 (54.4%) children were referred for further dental care; 91/124 (73.4%) attended any dental appointment. Attendance rates were 90% with a phone reminder system for appointments, attendance reduced when this system lapsed. CONCLUSIONS: Oral health is an important public health issue in refugee-background children, despite low levels of parent concern and very few parent reported caries. Routine direct oral health assessment is important in refugee-background children and co-ordinated health systems may help improve their attendance at dental services

Investigation of the diffusion behavior of sodium in Cu(In,Ga)Se₂ layers

Sodium diffusion in Cu(In,Ga)Se-2(CIGS) layers was investigated over a temperature range from 157 degrees C to 400 degrees C. The diffusion profiles were measured by secondary ion mass spectrometry. Sodium ions diffused from a sodium fluoride (NaF) layer on the CIGS surface into the CIGS layer. From Na diffusion profiles, the diffusion along grain boundaries could be distinguished from the diffusion into the grain interior. Atom-probe tomography measurements reveal that even at a low temperature of 157 degrees C bulk diffusion of sodium into CIGS occurs. Based on this data, the slower diffusion coefficient in the volume can be described by the Arrhenius equation D-Na(V) = 9.7 x 10(-9) exp(-0.36 eV/kBT) cm(2) s(-1) and the fast diffusion along the grain boundaries by D-Na(GB) = 6.5 x 10(-9) exp(-0.21 eV/kBT) cm 2 s(-1). Hence, we propose that sodium ions do not only passivate grain boundaries, but also act as dopants in the CIGS bulk. (C) 2014 AIP Publishing LL