Reliable and robust molecular sexing of the hen harrier (Circus cyaneus) using PCR-RFLP of the CHD1 gene

The hen harrier (Circus cyaneus) is a bird of prey that is persecuted in the United Kingdom, and there is a need for a DNA-based individual identification and sexing system for the use in forensic investigations. This study reports a new set of PCR primers for the chromo-helicase-DNA-binding protein 1 gene, which allows sexing using PCR-RFLP. Instead of exonic primers that amplify across a large intron, this set consists of a primer within the intron, enabling reduction in amplicon sizes from 356 to 212 bp and 565 to 219 bp in W and Z chromosomes. DNA degradation and dilution experiments demonstrate that this set is significantly more robust than one that amplifies across the intron, and sequencing of the intronic primer-binding region across several individuals shows that it is highly conserved. While our objective is to incorporate this primer set into an STR-based individualization kit, it may in the meantime prove useful in forensic or conservation studies

[\u3csup\u3e3\u3c/sup\u3eH]‑Raffinose, a Novel Radioactive Label for Determining Organ Sites of Catabolism of Proteins in the Circulation

The primary tissue sites of catabolism of plasma proteins with long circulating half-lives are unknown. It has been difficult to identify these sites because plasma proteins are delivered to tissues at relatively slow rates but are rapidly degraded intracellularly within lysosomes. Therefore, a tracer attached to protein is lost from the site of uptake before an amount sufficient for quantitation can accumulate. We hypothesized that sucrose (Glupal-2 /3Fruf) would be a useful label to circumvent this difficulty because of the stability of sucrose in lysosomes; and thus, sucrose should remain in tissue long after the protein to which it was attached had been degraded to products released from the lysosome. [G-3HJRaffinose (RAF, Galpal- Glupcwl- 2flFruf) was selected as the vehicle for attaching sucrose to protein. [31XjRAF was converted to the C-6 aldehydogalactose form with galactose oxidase and then covalently coupled to protein by reductive amination using NaBH3CN. [‘H]RAF was coupled first to two relatively long lived plasma proteins, bovine serum albumin and fetuin. The half-lives of these proteins in the rat circulation (&,* = -24 h) was unchanged, suggesting that RAF did not alter the normal mechanisms of protein clearance. When attached to short lived proteins with known sites of catabolism, such as asialofetuin, RNase B, and heatdenatured albumin, neither the tissue nor cellular sites of uptake of the proteins were altered. Thus, 13H]RAFasialofetuin was recovered almost exclusively (\u3e90%) in the liver parenchymal cell fraction, while both [JHJRAF-labeled RNase B and heat-denatured albumin were recovered primarily (\u3e85%) in nonparenchymal cells. In addition, the RAF label was observed to reside stably (tl,2 \u3e 100 h) in the liver following degradation of the carrier protein; in contrast, radioactivity from “‘Ilabeled asialofetuin or RNase B was rapidly (tl,z \u3c 30 min) lost from liver. Radioactivity from 13H]RAF-proteins was recovered in a lysosomally enriched subcellular fraction in liver and consisted of a low molecular weight species (-llOO), containing both glucose and fructose in a ratio similar to that in the original protein. The results of these studies establish that 13H]RAF useful radioactive tracer for detecting the tissue and cellular sites of catabolism of long lived circulating proteins

Fractional flow reserve vs. angiography in guiding management to optimize outcomes in non-ST-segment elevation myocardial infarction: the British Heart Foundation FAMOUS-NSTEMI randomized trial

Aim: We assessed the management and outcomes of non-ST segment elevation myocardial infarction (NSTEMI) patients randomly assigned to fractional flow reserve (FFR)-guided management or angiography-guided standard care. Methods and results: We conducted a prospective, multicentre, parallel group, 1 : 1 randomized, controlled trial in 350 NSTEMI patients with ≥ coronary stenosis ≥30% of the lumen diameter assessed visually (threshold for FFR measurement) (NCT01764334). Enrolment took place in six UK hospitals from October 2011 to May 2013. Fractional flow reserve was disclosed to the operator in the FFR-guided group (n = 176). Fractional flow reserve was measured but not disclosed in the angiography-guided group (n = 174). Fractional flow reserve ≤0.80 was an indication for revascularization by percutaneous coronary intervention (PCI) or coronary artery bypass surgery (CABG). The median (IQR) time from the index episode of myocardial ischaemia to angiography was 3 (2, 5) days. For the primary outcome, the proportion of patients treated initially by medical therapy was higher in the FFR-guided group than in the angiography-guided group [40 (22.7%) vs. 23 (13.2%), difference 95% (95% CI: 1.4%, 17.7%), P = 0.022]. Fractional flow reserve disclosure resulted in a change in treatment between medical therapy, PCI or CABG in 38 (21.6%) patients. At 12 months, revascularization remained lower in the FFR-guided group [79.0 vs. 86.8%, difference 7.8% (−0.2%, 15.8%), P = 0.054]. There were no statistically significant differences in health outcomes and quality of life between the groups. Conclusion: In NSTEMI patients, angiography-guided management was associated with higher rates of coronary revascularization compared with FFR-guided management. A larger trial is necessary to assess health outcomes and cost-effectiveness

Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform

Important insights into aging have been generated with the genetically tractable and short-lived budding yeast. However, it is still impossible today to continuously track cells by high-resolution microscopic imaging (e.g., fluorescent imaging) throughout their entire lifespan. Instead, the field still needs to rely on a 50-y-old laborious and time-consuming method to assess the lifespan of yeast cells and to isolate differentially aged cells for microscopic snapshots via manual dissection of daughter cells from the larger mother cell. Here, we are unique in achieving continuous and high-resolution microscopic imaging of the entire replicative lifespan of single yeast cells. Our microfluidic dissection platform features an optically prealigned single focal plane and an integrated array of soft elastomer-based micropads, used together to allow for trapping of mother cells, removal of daughter cells, monitoring gradual changes in aging, and unprecedented microscopic imaging of the whole aging process. Using the platform, we found remarkable age-associated changes in phenotypes (e.g., that cells can show strikingly differential cell and vacuole morphologies at the moment of their deaths), indicating substantial heterogeneity in cell aging and death. We envision the microfluidic dissection platform to become a major tool in aging research.

Dynamical Causes of the 2010/11 Texas–Northern Mexico Drought

The causes of the Texas–northern Mexico drought during 2010–11 are shown, using observations, reanalyses, and model simulations, to arise from a combination of ocean forcing and internal atmospheric variability. The drought began in fall 2010 and winter 2010/11 as a La Niña event developed in the tropical Pacific Ocean. Climate models forced by observed sea surface temperatures (SSTs) produced dry conditions in fall 2010 through spring 2011 associated with transient eddy moisture flux divergence related to a northward shift of the Pacific–North American storm track, typical of La Niña events. In contrast the observed drought was not associated with such a clear shift of the transient eddy fields and instead was significantly influenced by internal atmospheric variability including the negative North Atlantic Oscillation of winter 2010/11, which created mean flow moisture divergence and drying over the southern Plains and southeast United States. The models suggest that drought continuation into summer 2011 was not strongly SST forced. Mean flow circulation and moisture divergence anomalies were responsible for the summer 2011 drought, arising from either internal atmospheric variability or a response to dry summer soils not captured by the models. The summer of 2011 was one of the two driest and hottest summers over recent decades but it does not represent a clear outlier to the strong inverse relation between summer precipitation and temperature in the region. Seasonal forecasts at 3.5-month lead time did predict onset of the drought in fall and winter 2010/11 but not intensification into summer 2011, demonstrating the current, and likely inherent, inability to predict important aspects of North American droughts

Materials of the round table “Analytical methods and computer experiment in soft matter theory”

The paper contains materials of the discussion regarding the development, the problems of today and the prospects of analytical methods and computer simulations in soft matter theory which took place during the Workshop on modern problems of soft matter theory (August 27–31, 2000, Lviv, Ukraine).У статті подано матеріали дискусії щодо розвитку, сучасного стану та перспектив аналітичних методів і комп’ютерного моделювання в теоріі м’якої речовини, що відбулася підчас Наради із сучасних проблем теорії м’якої речовини (27–31 серпня 2000 р., м. Львів)

Active cooling control of the CLEO detector using a hydrocarbon coolant farm

We describe a novel approach to particle-detector cooling in which a modular farm of active coolant-control platforms provides independent and regulated heat removal from four recently upgraded subsystems of the CLEO detector: the ring-imaging Cherenkov detector, the drift chamber, the silicon vertex detector, and the beryllium beam pipe. We report on several aspects of the system: the suitability of using the aliphatic-hydrocarbon solvent PF(TM)-200IG as a heat-transfer fluid, the sensor elements and the mechanical design of the farm platforms, a control system that is founded upon a commercial programmable logic controller employed in industrial process-control applications, and a diagnostic system based on virtual instrumentation. We summarize the system's performance and point out the potential application of the design to future high-energy physics apparatus.Comment: 21 pages, LaTeX, 5 PostScript figures; version accepted for publication in Nuclear Instruments and Methods in Physics Research