Sexually transmitted infections and prostate cancer among men in the U.S. military

10.1158/1055-9965.EPI-08-1167Cancer Epidemiology Biomarkers and Prevention18102665-267

Automation of the ELISpot assay for high-throughput detection of antigen-specific T-cell responses

The enzyme linked immunospot (ELISpot) assay is a fundamental tool in cellular immunology, providing both quantitative and qualitative information on cellular cytokine responses to defined antigens. It enables the comprehensive screening of patient derived peripheral blood mononuclear cells to reveal the antigenic restriction of T-cell responses and is an emerging technique in clinical laboratory investigation of certain infectious diseases. As with all cellular-based assays, the final results of the assay are dependent on a number of technical variables that may impact precision if not highly standardised between operators. When studies that are large scale or using multiple antigens are set up manually, these assays may be labour intensive, have many manual handling steps, are subject to data and sample integrity failure and may show large inter-operator variability. Here we describe the successful automated performance of the interferon (IFN)-γ ELISpot assay from cell counting through to electronic capture of cytokine quantitation and present the results of a comparison between automated and manual performance of the ELISpot assay. The mean number of spot forming units enumerated by both methods for limiting dilutions of CMV, EBV and influenza (CEF)-derived peptides in six healthy individuals were highly correlated (r > 0.83, p < 0.05). The precision results from the automated system compared favourably with the manual ELISpot and further ensured electronic tracking, increased through-put and reduced turnaround time

<i>PIK3CA</i>-associated developmental disorders exhibit distinct classes of mutations with variable expression and tissue distribution.

Mosaicism is increasingly recognized as a cause of developmental disorders with the advent of next-generation sequencing (NGS). Mosaic mutations of &lt;i&gt;PIK3CA&lt;/i&gt; have been associated with the widest spectrum of phenotypes associated with overgrowth and vascular malformations. We performed targeted NGS using 2 independent deep-coverage methods that utilize molecular inversion probes and amplicon sequencing in a cohort of 241 samples from 181 individuals with brain and/or body overgrowth. We identified &lt;i&gt;PIK3CA&lt;/i&gt; mutations in 60 individuals. Several other individuals ( &lt;i&gt;n&lt;/i&gt; = 12) were identified separately to have mutations in &lt;i&gt;PIK3CA&lt;/i&gt; by clinical targeted-panel testing ( &lt;i&gt;n&lt;/i&gt; = 6), whole-exome sequencing ( &lt;i&gt;n&lt;/i&gt; = 5), or Sanger sequencing ( &lt;i&gt;n&lt;/i&gt; = 1). Based on the clinical and molecular features, this cohort segregated into three distinct groups: (a) severe focal overgrowth due to low-level but highly activating (hotspot) mutations, (b) predominantly brain overgrowth and less severe somatic overgrowth due to less-activating mutations, and (c) intermediate phenotypes (capillary malformations with overgrowth) with intermediately activating mutations. Sixteen of 29 &lt;i&gt;PIK3CA&lt;/i&gt; mutations were novel. We also identified constitutional &lt;i&gt;PIK3CA&lt;/i&gt; mutations in 10 patients. Our molecular data, combined with review of the literature, show that &lt;i&gt;PIK3CA&lt;/i&gt; -related overgrowth disorders comprise a discontinuous spectrum of disorders that correlate with the severity and distribution of mutations

Rights and responsibilities of individuals participating in medical research

Current geophysical knowledge of the planet Mercury is based upon observations from ground-based astronomy and flybys of the Mariner 10 spacecraft, along with theoretical and computational studies. Mercury has the highest uncompressed density of the terrestrial planets and by implication has a metallic core with a radius approximately 75% of the planetary radius. Mercury’s spin rate is stably locked at 1.5 times the orbital mean motion. Capture into this state is the natural result of tidal evolution if this is the only dissipative process affecting the spin, but the capture probability is enhanced if Mercury’s core were molten at the time of capture. The discovery of Mercury’s magnetic field by Mariner 10 suggests the possibility that the core is partially molten to the present, a result that is surprising given the planet’s size and a surface crater density indicative of early cessation of significant volcanic activity. A present-day liquid outer core within Mercury would require either a core sulfur content of at least several weight percent or an unusual history of heat loss from the planet’s core and silicate fraction. A crustal remanent contribution to Mercury’s observed magnetic field cannot be ruled out on the basis of current knowledge. Measurements from the MESSENGER orbiter, in combination with continued ground-based observations, hold the promise of setting on a firmer basis our understanding of the structure and evolution of Mercury’s interior and the relationship of that evolution to the planet’s geological history