Search

The circadian clock and the hypoxia signaling pathway are regulated by an integrated interplay of positive and negative feedback limbs that incorporate energy homeostasis and carcinogenesis. We show that the negative circadian regulator CRY1 is also a negative regulator of hypoxiainducible factor (HIF). Mechanistically, CRY1 interacts with the basic helixloop-helix domain of HIF-1α via its tail region. Subsequently, CRY1 reduces HIF-1α half-life and binding of HIFs to target gene promoters. This appeared to be CRY1 specific since genetic disruption of CRY1 but not CRY2 affected the hypoxia response. Further, CRY1-deficiency could induce cellular HIF levels, proliferation and migration, which could be reversed by CRISPR/Cas9 or shRNA mediated HIF knock-out. Altogether, our study provides a mechanistic explanation for genetic association studies linking a disruption of the circadian clock with hypoxia-associated processes such as carcinogenesis.

Background Ageing is characteristically accompanied by changes in vascular endothelial markers and growth factor as well as increased cellular death. We analysed the associations of the plasma levels of vascular cell adhesion molecule-1 (VCAM-1), hepatocyte growth factor (HGF) and soluble Fas (sFAS), and their combinations, with 4-year mortality to identify new biomarkers. Methods A total of 238 individuals, both community-dwelling and institutionalised, aged 89‒91 years and participating in the Vitality 90 + study were included. Biomarkers of endothelial function (VCAM-1), growth factor (HGF) and a marker of apoptosis (sFAS) were determined from plasma using Luminex® technology. This newly-determined data was combined with earlier data, e.g., 4-year mortality and medical history. Results Subjects who died during the follow-up had higher baseline plasma levels of VCAM-1, sFas, and HGF. When other known risk factors were adjusted for, subjects in the highest concentration tertile for VCAM-1 (HR 1.85; 95% CI, 1.12–3.05) and HGF (HR 2.22; 95% CI, 1.33–3.71) had higher mortality compared to those in the lowest tertile. In the adjusted analyses, when compared to subjects with none of the biomarkers in the highest concentration tertile, mortality was also higher when sFas and VCAM-1 were simultaneously (HR 2.03; 95% CI, 1.13–3.64) or all three were simultaneously (HR 3.63; 95% CI, 1.65–7.97) in the highest concentration tertile. Conclusions Our results suggest that increased concentrations of these biomarkers, separately and in combination, associate with mortality among the aged and are prognostic markers of death.

We thank Weingartner et al. for their thoughtful comments on our report [(1)][1]. The current guidelines for the management of dyslipidemia endorsed by the European Atherosclerosis Society (EAS) and the European Society of Cardiology (ESC) do not include a clear recommendation for the use of

Top of pageAbstract The post-genomic era necessitates new tools to decode the vast nucleotide sequence data into biologically relevant knowledge. This problem has created a new field in molecular biology and genetics called functional genomics, which addresses the functions of genes and gene products at the level of whole organisms. Viral vector systems have been explored as one solution to solve these problems. We have constructed a new baculovirus expression vector system (BEVS) based tetra-promoter vector (pBVboostFG) that enables screening of gene or cDNA libraries for functional genomic studies. The vector enables an all-in-one strategy for gene expression in mammalian, insect and bacterial cells. In addition, it is also suitable for direct use in vivo in gene therapy. Recombinant virus preparation is based on an improved mini Tn7 transpositional system that allows straightforward and fast production of baculoviruses with high diversity and negligible background. Genes or cDNAs were cloned into pBVboostFG using the bacteriophage lambda's recombination system and production of 20 different proteins was studied in bacteria, insect and mammalian cells. As an example of using the vector for library screening, a chromophoric Thr65-Tyr-Gly67-stretch of EGFP was destroyed and subsequently restored by PCR and the EGFP library was screened for the color variants. All studied proteins were efficiently produced using pBVboostFG system and with different expression hosts. In addition, three different fluorescent protein variants were screened out from the generated chromophore library. Finally, the BVboostFG vector was used without any additional subcloning steps to efficiently transduce choroid plexus epithelial cells in rat brain in vivo. The pBVboostFG system enables easy and efficient production of desired proteins or screening of genome-wide libraries in multiple cell types with one-step cloning. This makes it an efficient new platform technology for functional genomics and gene therapy.