Involvement of global genome repair, transcription coupled repair, and chromatin remodeling in UV DNA damage response changes during development

Nucleotide Excision Repair (NER), which removes a variety of helix-distorting lesions from DNA, is initiated by two distinct DNA damage-sensing mechanisms. Transcription Coupled Repair (TCR) removes damage from the active strand of transcribed genes and depends on the SWI/SNF family protein CSB. Global Genome Repair (GGR) removes damage present elsewhere in the genome and depends on damage recognition by the XPC/RAD23/Centrin2 complex. Currently, it is not well understood to what extent both pathways contribute to genome maintenance and cell survival in a developing organism exposed to UV light. Here, we show that eukaryotic NER, initiated by two distinct subpathways, is well conserved in the nematode Caenorhabditis elegans. In C. elegans, involvement of TCR and GGR in the UV-induced DNA damage response changes during development. In germ cells and early embryos, we find that GGR is the major pathway contributing to normal development and survival after UV irradiation, whereas in later developmental stages TCR is predominantly engaged. Furthermore, we identify four ISWI/Cohesin and four SWI/SNF family chromatin remodeling factors that are implicated in the UV damage response in a developmental stage dependent manner. These in vivo studies strongly suggest that involvement of different repair pathways and chromatin remodeling proteins in UV-induced DNA repair depends on developmental stage of cells

Prednisolone induces the Wnt signalling pathway in 3T3-L1 adipocytes

Contains fulltext : 117166.pdf (publisher's version ) (Open Access)Synthetic glucocorticoids are potent anti-inflammatory drugs but show dose-dependent metabolic side effects such as the development of insulin resistance and obesity. The precise mechanisms involved in these glucocorticoid-induced side effects, and especially the participation of adipose tissue in this are not completely understood. We used a combination of transcriptomics, antibody arrays and bioinformatics approaches to characterize prednisolone-induced alterations in gene expression and adipokine secretion, which could underlie metabolic dysfunction in 3T3-L1 adipocytes. Several pathways, including cytokine signalling, Akt signalling, and Wnt signalling were found to be regulated at multiple levels, showing that these processes are targeted by prednisolone. These results suggest that mechanisms by which prednisolone induce insulin resistance include dysregulation of wnt signalling and immune response processes. These pathways may provide interesting targets for the development of improved glucocorticoids

Drug prescriptions unadapted to the renal function in patients aged 80 years and older (a.)

Background: Drug-related problems are common in older people. Often they are related to low estimated glomerular filtration rate (eGFR), which has a high prevalence among older adults. The aim of this study was to investigate inappropriate drug prescriptions and dose adaptations in a very old population and their relationship with the eGFR. Method: Design: A cross-sectional study within a Belgian prospective population-based cohort study (the BELFRAIL study) of 539 participants aged 80 years and older (mean age 85 years). Drug prescriptions at inclusion were reported by the participant's responsible general practitioner. The eGFR was estimated using the MDRD equation. Based on their eGFR, the participants were divided in three groups: > 50, 30-50 and < 30 ml/min/1.73 m², respectively. Drug prescriptions were analysed in different eGFR groups. The prevalence and odds ratios of inappropriate drugs and the unadjusted defined daily doses (DDD) of the participant eGFRs were calculated. Results: Thirty-six (of 111) and eight (of 31) of the participants with an eGFR between 30-50 and < 30 ml/min/1.73 m², respectively, had at least one inappropriate drug prescribed. No decrease in mean DDD, was observed in any prescribed drug in both lower eGFR groups. Participants with a lower eGFR were at higher risk of receiving gliclazide (OR: 4.51; 95% CI: 1.45-14.02) or unadjusted doses of allopurinol (OR: 3.48; 95% CI: 1.26-9.61). Conclusion: Drug prescriptions inappropriate for patient eGFR are common in subjects aged 80 years and older, despite automatic eGFR reporting

A HPLC-based glycoanalytical protocol allows the use of natural O-glycans derived from glycoproteins as substrates for glycosidase discovery from microbial culture

Many disorders are characterised by changes in O-glycosylation, but analysis of O-glycosylation has been limited by the availability of specific endo- and exo-glycosidases. As a result chemical methods are employed. However, these may give rise to glycan degradation, so therefore novel O-glycosidases are needed. Artificial substrates do not always identify every glycosidase activity present in an extract. To overcome this, an HPLC-based protocol for glycosidase identification from microbial culture was developed using natural O-glycans and O-glycosylated glycoproteins (porcine stomach mucin and fetuin) as substrates. O-glycans were released by ammonia-based β-elimination for use as substrates, and the bacterial culture supernatants were subjected to ultrafiltration to separate the proteins from glycans and low molecular size molecules. Two bacterial cultures, the psychrotroph Arthrobacter C1-1 and a Corynebacterium isolate, were examined as potential sources of novel glycosidases. Arthrobacter C1-1 culture contained a β-galactosidase and N-acetyl-β-glucosaminidase when assayed using 4-methylumbelliferyl substrates, but when defucosylated O-glycans from porcine stomach mucin were used as substrate, the extract did not cleave β-linked galactose or N-acetylglucosamine. Sialidase activity was identified in Corynebacterium culture supernatant, which hydrolysed sialic acid from fetuin glycans. When both culture supernatants were assayed using the glycoproteins as substrate, neither contained endoglycosidase activity. This method may be applied to investigate a microbial or other extract for glycosidase activity, and has potential for scale-up on high-throughput platforms