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Preliminary results for the extraction and measurement of cosmogenic in situ C-14 from quartz
Radiocarbon is produced within minerals at the earth's surface (in situ production) by a number of spallation reactions. Its relatively short half-life of 5730 yr provides us with a unique cosmogenic nuclide tool for the measurement of rapid erosion rates ( gt 10(-3) cm yr(-1)) and events occurring over the past 25 kyr. At SUERC, we have designed and built a vacuum system to extract C-14 from quartz which is based on a system developed at the University of Arizona. This system uses resistance heating of samples to a temperature of approximately 1100 degreesC in the presence of lithium metaborate(LiBO2) to dissolve the quartz and liberate any carbon present. During extraction, the carbon is oxidized to CO2 in an O-2 atmosphere so that it may be collected cryogenically. The CO2 is Subsequently purified and converted to graphite for accelerator mass spectrometry (AMS) measurement. One of the biggest problems in measuring in situ C-14 is establishing a low and reproducible system blank and efficient extraction of the in situ C-14 Component. Here, we present initial data for C-14- free CO2, derived from geological carbonate and added to the vacuum system to determine the system blank. Shielded quartz samples (which should be C-14 free) and a surface quartz sample routinely analyzed at the University of Arizona were also analyzed at SUERC, and the data compared with values derived from the University of Arizona system
<em>DAAM2</em> variants cause nephrotic syndrome via actin dysregulation.
The discovery of >60 monogenic causes of nephrotic syndrome (NS) has revealed a central role for the actin regulators RhoA/Rac1/Cdc42 and their effectors, including the formin INF2. By whole-exome sequencing (WES), we here discovered bi-allelic variants in the formin DAAM2 in four unrelated families with steroid-resistant NS. We show that DAAM2 localizes to the cytoplasm in podocytes and in kidney sections. Further, the variants impair DAAM2-dependent actin remodeling processes: wild-type DAAM2 cDNA, but not cDNA representing missense variants found in individuals with NS, rescued reduced podocyte migration rate (PMR) and restored reduced filopodia formation in shRNA-induced DAAM2-knockdown podocytes. Filopodia restoration was also induced by the formin-activating molecule IMM-01. DAAM2 also co-localizes and co-immunoprecipitates with INF2, which is intriguing since variants in both formins cause NS. Using in vitro bulk and TIRF microscopy assays, we find that DAAM2 variants alter actin assembly activities of the formin. In a Xenopus daam2-CRISPR knockout model, we demonstrate actin dysregulation in vivo and glomerular maldevelopment that is rescued by WT-DAAM2 mRNA. We conclude that DAAM2 variants are a likely cause of monogenic human SRNS due to actin dysregulation in podocytes. Further, we provide evidence that DAAM2-associated SRNS may be amenable to treatment using actin regulating compounds
Whole-Exome Sequencing Identifies Causative Mutations in Families with Congenital Anomalies of the Kidney and Urinary Tract.
Item does not contain fulltextBACKGROUND: Congenital anomalies of the kidney and urinary tract (CAKUT) are the most prevalent cause of kidney disease in the first three decades of life. Previous gene panel studies showed monogenic causation in up to 12% of patients with CAKUT. METHODS: We applied whole-exome sequencing to analyze the genotypes of individuals from 232 families with CAKUT, evaluating for mutations in single genes known to cause human CAKUT and genes known to cause CAKUT in mice. In consanguineous or multiplex families, we additionally performed a search for novel monogenic causes of CAKUT. RESULTS: In 29 families (13%), we detected a causative mutation in a known gene for isolated or syndromic CAKUT that sufficiently explained the patient's CAKUT phenotype. In three families (1%), we detected a mutation in a gene reported to cause a phenocopy of CAKUT. In 15 of 155 families with isolated CAKUT, we detected deleterious mutations in syndromic CAKUT genes. Our additional search for novel monogenic causes of CAKUT in consanguineous and multiplex families revealed a potential single, novel monogenic CAKUT gene in 19 of 232 families (8%). CONCLUSIONS: We identified monogenic mutations in a known human CAKUT gene or CAKUT phenocopy gene as the cause of disease in 14% of the CAKUT families in this study. Whole-exome sequencing provides an etiologic diagnosis in a high fraction of patients with CAKUT and will provide a new basis for the mechanistic understanding of CAKUT.1 september 201