The Caenorhabditis elegans Gene mfap-1 Encodes a Nuclear Protein That Affects Alternative Splicing

RNA splicing is a major regulatory mechanism for controlling eukaryotic gene expression. By generating various splice isoforms from a single pre–mRNA, alternative splicing plays a key role in promoting the evolving complexity of metazoans. Numerous splicing factors have been identified. However, the in vivo functions of many splicing factors remain to be understood. In vivo studies are essential for understanding the molecular mechanisms of RNA splicing and the biology of numerous RNA splicing-related diseases. We previously isolated a Caenorhabditis elegans mutant defective in an essential gene from a genetic screen for suppressors of the rubberband Unc phenotype of unc-93(e1500) animals. This mutant contains missense mutations in two adjacent codons of the C. elegans microfibrillar-associated protein 1 gene mfap-1. mfap-1(n4564 n5214) suppresses the Unc phenotypes of different rubberband Unc mutants in a pattern similar to that of mutations in the splicing factor genes uaf-1 (the C. elegans U2AF large subunit gene) and sfa-1 (the C. elegans SF1/BBP gene). We used the endogenous gene tos-1 as a reporter for splicing and detected increased intron 1 retention and exon 3 skipping of tos-1 transcripts in mfap-1(n4564 n5214) animals. Using a yeast two-hybrid screen, we isolated splicing factors as potential MFAP-1 interactors. Our studies indicate that C. elegans mfap-1 encodes a splicing factor that can affect alternative splicing.National Natural Science Foundation (China) (Grant 30971639)United States. National Institutes of Health (Grant GM24663

Direct activation of NADPH oxidase 2 by 2-deoxyribose-1-phosphate triggers nuclear factor kappa B-dependent angiogenesis

This is the author accepted manuscript. The final version is available from Mary Ann Liebert via the DOI in this recordAims: Deoxyribose-1-phosphate (dRP) is a pro-angiogenic paracrine stimulus released by cancer cells, platelets and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, electron paramagnetic resonance (EPR), pull-down of dRPinteracting proteins followed by immunoblotting, gene silencing of different NOXs and their regulatory co-subunits by siRNA transfection, and experiments with inhibitors of the sugar transporter GLUT1 were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NADPH oxidase 2 (NOX2) complex, but not NOX4. Increased reactive oxygen species (ROS) generation in response to NOX2 activity leads to redoxdependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces VEGF receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and the NOX2-dependent stimulation of angiogenesis by dRP was confirmed in excisional wound and matrigel plug vascularisation assays in vivo using NOX2-/- mice. Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex. Conclusions: This study describes a novel molecular mechanism underlying the pro-angiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and the upregulation of VEGFR2This work was sponsored by the Biotechnology and Biological Sciences Research Council (BB/J002690/1), the Medical Research Council (MRC/CIC/2015) and the British Heart Foundation (PG/15/40/31522

Disparities exist between National food group recommendations and the dietary intakes of women

<p>Abstract</p> <p>Background</p> <p>Preconception and pregnancy dietary intakes can influence the health of future generations. In this study we compared the food intakes of reproductive-aged women by pregnancy status, to current Australian recommendations.</p> <p>Methods</p> <p>Data are from the Australian Longitudinal Study on Women's Health, younger cohort aged 25-30 years in 2003, with self-reported status as pregnant (n = 606), trying to conceive (n = 454), given birth in the last 12 months (n = 829) or other (n = 5597). Diet was assessed using a validated 74-item food frequency questionnaire. Food group servings and nutrient intakes were compared to the Australian Guide to Healthy Eating (AGHE) and Australian Nutrient Reference Values (NRVs).</p> <p>Results</p> <p>No women met all AGHE food group recommendations. Highest adherence rates [mean (95% CI) servings/day] were for meat [85%, 1.9(1.8-1.9)], fruit [44%, 2.1(2.1-2.2)] and dairy [35%, 1.8(1.8-1.9)], with < 14% meeting remaining recommendations. Women who achieved NRVs (folate, iron, calcium, zinc, fibre) for pregnancy, breastfeeding and adult life stages were 1.5%, 3.3% and 13.7%, respectively. Compared to AGHE, women consumed more servings of fruit (4.9 vs 4.0;<it>P </it>= 0.034) and dairy (3.4 vs 2.0;<it>P </it>= 0.006) to achieve pregnancy NRVs; more dairy (2.9 vs 2.0;<it>P </it>= 0.001), less fruit (3.9 vs 5.0;<it>P </it>< .001) and vegetables (3.4 vs 7.0;<it>P </it>< .001) to achieve breastfeeding NRVs; more fruit (3.6 vs 3.0;<it>P </it>< .001), dairy (2.5 vs 2.0;<it>P </it>< .001), meat (1.8 vs 1.5;<it>P </it>= 0.015), less vegetables (3.6 vs 5.0;<it>P </it>< .001) to achieve adult NRVs.</p> <p>Conclusions</p> <p>The AGHE does not align with contemporary diets of Australian women or enable them to meet all NRVs. Current tools to guide food consumption by women during pregnancy require revision.</p