Diversity in parasitic nematode genomes: the microRNAs of Brugia pahangi and Haemonchus contortus are largely novel

<b>BACKGROUND:</b> MicroRNAs (miRNAs) play key roles in regulating post-transcriptional gene expression and are essential for development in the free-living nematode Caenorhabditis elegans and in higher organisms. Whether microRNAs are involved in regulating developmental programs of parasitic nematodes is currently unknown. Here we describe the the miRNA repertoire of two important parasitic nematodes as an essential first step in addressing this question. <b>RESULTS:</b> The small RNAs from larval and adult stages of two parasitic species, Brugia pahangi and Haemonchus contortus, were identified using deep-sequencing and bioinformatic approaches. Comparative analysis to known miRNA sequences reveals that the majority of these miRNAs are novel. Some novel miRNAs are abundantly expressed and display developmental regulation, suggesting important functional roles. Despite the lack of conservation in the miRNA repertoire, genomic positioning of certain miRNAs within or close to specific coding genes is remarkably conserved across diverse species, indicating selection for these associations. Endogenous small-interfering RNAs and Piwi-interacting (pi)RNAs, which regulate gene and transposon expression, were also identified. piRNAs are expressed in adult stage H. contortus, supporting a conserved role in germline maintenance in some parasitic nematodes. <b>CONCLUSIONS:</b> This in-depth comparative analysis of nematode miRNAs reveals the high level of divergence across species and identifies novel sequences potentially involved in development. Expression of novel miRNAs may reflect adaptations to different environments and lifestyles. Our findings provide a detailed foundation for further study of the evolution and function of miRNAs within nematodes and for identifying potential targets for intervention

A Novel Quantitative Simple Brain Metric Using MR Imaging for Preterm Infants

BACKGROUND AND PURPOSE: The application of volumetric techniques to preterm infants has revealed brain volume reductions. Such quantitative data are not available in routine neonatal radiologic care. The objective of this study was to develop simple brain metrics to compare brain size in preterm and term infants and to correlate these metrics with brain volumes from volumetric MR imaging techniques.MATERIALS AND METHODS: MR images from 189 preterm infants <30 weeks’ gestational age or <1250 g birthweight scanned at term-equivalent age and 36 term infants were studied. Fifteen tissue and fluid measures were systematically evaluated on 4 selected sections. The results were correlated with total brain, gray matter, white matter, and CSF volumes. RESULTS: The mean bifrontal, biparietal, and transverse cerebellar diameters were reduced (−11.6%, 95% confidence interval [CI], −13.8% to −9.3%; −12%, 95% CI, −14% to −9.8%; and −8.7%, 95% CI, −10.5% to −7% respectively) and the mean left ventricle diameter was increased (+22.3%, 95% CI, 2.9%–41.6%) in preterm infants (P < .01). Strong correlations were found between the bifrontal and biparietal measures with total brain tissue volume, whereas the size of the ventricles and the interhemispheric measure correlated with CSF volume. Intraobserver reliability was high (intraclass correlation coefficients [ICC], >0.7), where interobserver agreement was acceptable for tissue measures (ICC, >0.6) but lower for fluid measures (ICC, <0.4) CONCLUSIONS: Simple brain metrics at term-equivalent age showed smaller brain diameters and increased ventricle size in preterm infants compared with full-term infants. These measures represent a reliable and easily applicable method to quantify brain growth and assess brain atrophy in this at-risk population

A microflow cytometer for microsphere-based immunoassays using integrated optics and inertial particle focussing

We present work towards a microflow cytometer for performing multiplex immunoassays using commercially available fluorescently-labelled microspheres. The device consists of a silica chip with integrated GeO2:SiO2 channel waveguides which deliver excitation light orthogonally to an etched flow channel [1], [2]. The rectangular cross section, 2:1 aspect ratio flow channel and flow rate create an inertial focussing effect on the microspheres [3] which ensures they flow through the plane of maximum optical excitation, halfway up the height of the channel, with minimal positional variation. The optical waveguide core is fabricated by magnetron sputtering of GeO2:SiO2 films which are then etched to form channel waveguides by ICP etching. The silica cladding, up to 13.5 µm thick, is deposited by either flame hydrolysis deposition or a combination of magnetron sputtering followed by PECVD. Fluidic channels are etched with ICP etching. Channels with the dimensions of 14.1 µm x 27.5 µm and near vertical sidewalls (91°±4°) have been produced in silica as shown in the cross section in Figure 1A. Figure 1B shows a device with the fluidic channel etched through waveguides clad with PECVD silica. Design parameters were established with PDMS test channels 25.5 µm deep by 12.2 µm wide. Figures 2A and 2B show transmission fluorescence imaging of streaks from multiple 5.6µm diameter microspheres flowing at 0.49 m/s down the fluidic channel. The microspheres are shown to be focused into a tight stream at 15 mm from the channel entrance in Figure 2C, indicating the minimum channel length required for the final devices. Future work will include dual channel quantification of microsphere fluorescence and development of an assay for TNFalpha and later multiplex measurements. Collection of fluorescence with channel waveguides and also characterisation of transmission measurements from flowing microspheres will also be studied

Anticoagulant rodenticides in red kites (Milvus milvus) in Britain in 2017 and 2018

Second generation anticoagulant rodenticides (SGARs) can be toxic to all mammals and birds. Various studies have shown that, in Britain, there is widespread exposure to SGARs in a diverse range of predatory mammals and birds, including red kites (Milvus milvus) which scavenge dead rats, a target species for rodent control. The Wildlife Incident Monitoring Scheme (WIIS) and the Predatory Bird Monitoring Scheme (PBMS) have shown that some mortalities result from this secondary exposure. In the present study, we analysed liver SGAR residues in 77 red kites that had been found dead in Britain in either 2017 or 2018. The carcasses were submitted to and necropsied by the Disease Risk Analysis and Health Surveillance (DRAHS) programme, the PBMS, the WIIS for England & Wales, the WIIS for Scotland and the Raptor Health Scotland study; the livers from the kites were subsequently analysed for SGAR residues. All the organisations are partners in the WILDCOMS network that promotes collaboration among surveillance schemes that monitor disease and contaminants in vertebrate wildlife. All of the 66 kites from England & Wales and 10 of the 11 red kites from Scotland had detectable liver residues of at least one SGAR. When considering the sample of kites as a whole, brodifacoum, difenacoum and bromadiolone were each detected in 73, 71 and 60 kites, respectively. Difethialone was found in 11 individuals while flocoumafen was detected in only one bird. Sum liver SGAR concentrations ranged between non-detected and 1218 ng/g wet wt. (arithmetic mean: 246 ng/g, median 154 ng/g). Post-mortem examinations indicated that 13 (16.8%) of red kites examined had internal haemorrhaging that was not associated with detectable trauma and had detectable liver SGAR concentrations. These birds had sum SGAR liver concentrations that ranged from 135 ng/g wet weight to 1218 ng/g wet weight. SGARs were considered a contributory cause of death in these cases. The stewardship scheme for anticoagulant rodenticides came fully into force in mid-2016 as re-registration of products for use in the UK was completed. A key aim is to reduce exposure of non-target wildlife to anticoagulant rodenticides but stewardship also aims to maintain efficacious rat control and so the number and density of AR-contaminated rats may remain unchanged. However, diligent searching, removal and safe disposal of poisoned rats, as promoted by stewardship, might be expected to reduce the availability of poisoned dead rats to red kites [and other scavengers] and thereby reduce the proportion of birds that are exposed and/or the magnitude of exposure. Concomitant with stewardship was a relaxation of the indoor use only restriction previously applied to brodifacoum, flocoumafen and difethialone, the three most acutely toxic SGARs. Any consequent increase in outdoor use of these three SGARs could increase the risk of secondary exposure in red kites. We therefore compared the data in the current report with that collected in 2015 and 2016 to determine if there was any evidence of a change in pattern or magnitude of exposure in red kites that might be connected to stewardship and/or change in usage restriction. The proportion of red kites exposed to SGARs in 2015 (90.6%), 2016 (89.6%) 2017 (96,4%) and 2018 (100%) was always 90% or more; the higher percentages in 2017 and 2018 were principally due to a greater proportion of birds from Scotland containing residues. Brodifacoum and difenacoum were the most prevalent compounds (89% of red kites across the four years for each compound) along with bromadiolone (75%). On average, there were residues of three different SGARs in each kite liver. There was no significant difference between years in liver sum (Σ) SGAR concentrations. We investigated if there was a change between years in the exposure of red kites to brodifacoum, flocoumafen and difethialone, the compounds for which indoor only usage restrictions were relaxed in 2016. To enable statistical analysis of data on residue prevalence, it was necessary to pool the data into two-year blocks. Data on presence/absence of detectable brodifacoum, flocoumafen or difethialone residues were therefore compared for 2015/16 (pre and year of implementation of change in usage restriction) and 2017/18 (post-change in usage restriction). The proportion of red kites with detectable residues was 82% (50 out of 61 red kites) in 2015/16 but significantly higher (95%; 73 out of 77 red kites) in 2017/18. However, there was also an increase [albeit not statistically significant] in the proportion of red kites with detectable liver difenacoum or bromadiolone residues (90% in 2015/16 vs. 97% in 2017/18). Therefore, these data may simply reflect an increase in the prevalence of exposure to SGARs generally rather than any effect of change in usage restriction. There was no difference between the four years in the summed magnitude of liver brodifacoum, difethialone and flocoumafen concentrations. The percentage of red kites examined that were diagnosed as birds in which SGARs were implicated as a contributory cause of death did not differ significantly between individual years nor show a significant trend across the years; the overall average across the four years was 22%. However, if data were pooled by pairs of years (2017/8 vs 2015/16), the proportion of red kites for which SGARs were implicated as a contributory cause of death was lower (18%) in 2017/18 than in 2015/16 (33%) for red kites from England & Wales. Our findings do not indicate that there has been any reduction in exposure in red kites to SGARs following implementation of stewardship, in terms of either the proportion of individuals exposed or the magnitude of residues detected. There is some evidence (depending upon the statistical approach used) that the proportion of red kites in which SGARs were implicated as a contributory mortality factor has decreased in more recent years. There was no clear evidence that relaxation of usage restrictions on brodifacoum, difethialone and flocoumafen has altered the pattern of residue accumulation in red kites to date

Localized f electrons in CexLa1-xRhIn5: dHvA Measurements

Measurements of the de Haas-van Alphen effect in CexLa1-xRhIn5 reveal that the Ce 4f electrons remain localized for all x, with the mass enhancement and progressive loss of one spin from the de Haas-van Alphen signal resulting from spin fluctuation effects. This behavior may be typical of antiferromagnetic heavy fermion compounds, inspite of the fact that the 4f electron localization in CeRhIn5 is driven, in part, by a spin-density wave instability.Comment: 4 pages, 4 figures, submitted to PR

Time Dependence of Tip Morphology during Cellular/Dendritic Arrayed Growth

Succinonitrile-1.9 wt pct acetone has been directionally solidified in 0.7 X 0.7-cm-square cross section pyrex ampoules in order to observe the cell/dendrite tip morphologies, not influenced by the 'wall effects', which are present during growth in the generally used thin (about 200 gm) crucibles. The tips do not maintain a steady-state shape, as is generally assumed. Instead, they fluctuate within a shape envelope. The extent of fluctuation increases with decreasing growth speed, as the micro structure changes from the dendritic to cellular. The influence of natural convection has been examined by comparing these morphologies with those grown, without convection, in the thin ampoules

S. cerevisiae Srs2 helicase ensures normal recombination intermediate metabolism during meiosis and prevents accumulation of Rad51 aggregates

YesWe investigated the meiotic role of Srs2, a multi-functional DNA helicase/translocase that destabilises Rad51-DNA filaments and is thought to regulate strand invasion and prevent hyper-recombination during the mitotic cell cycle. We find that Srs2 activity is required for normal meiotic progression and spore viability. A significant fraction of srs2 mutant cells progress through both meiotic divisions without separating the bulk of their chromatin, although in such cells sister centromeres often separate. Undivided nuclei contain aggregates of Rad51 colocalised with the ssDNA-binding protein RPA, suggesting the presence of persistent single-strand DNA. Rad51 aggregate formation requires Spo11-induced DSBs, Rad51 strand-invasion activity and progression past the pachytene stage of meiosis, but not the DSB end-resection or the bias towards interhomologue strand invasion characteristic of normal meiosis. srs2 mutants also display altered meiotic recombination intermediate metabolism, revealed by defects in the formation of stable joint molecules. We suggest that Srs2, by limiting Rad51 accumulation on DNA, prevents the formation of aberrant recombination intermediates that otherwise would persist and interfere with normal chromosome segregation and nuclear division.Biotechnology and Biological Sciences Research Council (BB/K009346/1

Women, men and coronary heart disease: a review of the qualitative literature

Aim. This paper presents a review of the qualitative literature which examines the experiences of patients with coronary heart disease. The paper also assesses whether the experiences of both female and male patients are reflected in the literature and summarizes key themes. Background. Understanding patients' experiences of their illness is important for coronary heart disease prevention and education. Qualitative methods are particularly suited to eliciting patients' detailed understandings and perceptions of illness. As much previous research has been 'gender neutral', this review pays particular attention to gender. Methods. Published papers from 60 qualitative studies were identified for the review through searches in MEDLINE, EMBASE, CINAHL, PREMEDLINE, PsychINFO, Social Sciences Citation Index and Web of Science using keywords related to coronary heart disease. Findings. Early qualitative studies of patients with coronary heart disease were conducted almost exclusively with men, and tended to generalize from 'male' experience to 'human' experience. By the late 1990s this pattern had changed, with the majority of studies including women and many being conducted with solely female samples. However, many studies that include both male and female coronary heart disease patients still do not have a specific gender focus. Key themes in the literature include interpreting symptoms and seeking help, belief about coronary 'candidates' and relationships with health professionals. The influence of social roles is important: many female patients have difficulties reconciling family responsibilities and medical advice, while male patients worry about being absent from work. Conclusions. There is a need for studies that compare the experiences of men and women. There is also an urgent need for work that takes masculinity and gender roles into account when exploring the experiences of men with coronary heart disease

Eutectic colony formation: A phase field study

Eutectic two-phase cells, also known as eutectic colonies, are commonly observed during the solidification of ternary alloys when the composition is close to a binary eutectic valley. In analogy with the solidification cells formed in dilute binary alloys, colony formation is triggered by a morphological instability of a macroscopically planar eutectic solidification front due to the rejection by both solid phases of a ternary impurity that diffuses in the liquid. Here we develop a phase-field model of a binary eutectic with a dilute ternary impurity and we investigate by dynamical simulations both the initial linear regime of this instability, and the subsequent highly nonlinear evolution of the interface that leads to fully developed two-phase cells with a spacing much larger than the lamellar spacing. We find a good overall agreement with our recent linear stability analysis [M. Plapp and A. Karma, Phys. Rev. E 60, 6865 (1999)], which predicts a destabilization of the front by long-wavelength modes that may be stationary or oscillatory. A fine comparison, however, reveals that the assumption commonly attributed to Cahn that lamella grow perpendicular to the envelope of the solidification front is weakly violated in the phase-field simulations. We show that, even though weak, this violation has an important quantitative effect on the stability properties of the eutectic front. We also investigate the dynamics of fully developed colonies and find that the large-scale envelope of the composite eutectic front does not converge to a steady state, but exhibits cell elimination and tip-splitting events up to the largest times simulated.Comment: 18 pages, 18 EPS figures, RevTeX twocolumn, submitted to Phys. Rev.