A simple serum depletion method for proteomics analysis

Serum is the body fluid most often used in biomarker discovery. Albumin, the most abundant serum protein, contributes approximately 50% of the serum protein content, with an additional dozen abundant proteins dominating the rest of the serum proteome. To profile this challenging protein mixture by proteomics, the abundant proteins must be depleted to allow for detection of the low-abundant proteins, the primary biomarker targets. Current serum depletion approaches for proteomics are costly and relatively complex to couple with protein digestion. We demonstrate a simple, affordable serum depletion methodology that, within a few minutes of processing, results in two captured serum fractions – albumin-depleted and albumin-rich – which are digested in situ. We believe our method is a useful addition to the biomarker sample preparation toolbox

A simplified, cost- and time-effective procedure for genotyping pearl millet in resource-limited laboratories

Procedures for DNA extraction and genotyping of large plant populations are cumbersome and expensive for resource-limited laboratories. Through eliminating or changing several steps used in DNA extraction, PCR amplification and PAGE electrophoresis in pearl millet [Pennisetum glaucum (L.) R. Br.], we developed a modified procedure that reduced the cost of consumables and required less time without compromising data quality. In the revised procedure, DNA was extracted by incubating 0.5 - 0.7 g ground young leaf tissue in 2% CTAB/-mercaptoethanol followed by refrigerated differential centrifugations with phenol:chloroform:isoamylalcohol. Steps such as additional  phenol/chloroform treatments, DNA pellet drying followed by RNase treatments and incubation were eliminated, reducing use of costly and corrosive chemicals and saving time. DNA produced from 174 genotypes exhibited an average concentration of 640 ng/ìL and average optical density ratio of 1.9. PCR amplification of SSR markers with this DNA produced clear and scorable bands following ethidium bromide stained agarose and silver stained polyacrylamide gel eletcrophoresis. Post PCR duplexing of two or more microsatellites based on different lengths of base pairs reduced the time and cost per unit data generation by up to half as compared to single marker per PAGE. Cluster analysis performed on the marker data generated 11 SSR primers following these procedures formed two main groups from genotypes of the U.S. origin. In summary, the procedures reported are simplified, shortened and economical and well suited for resource limited laboratories engaged in molecular breeding requiring large volume of genotyping

Assessing the quality of antenatal corticosteroids in low- and middle-income countries: A systematic review.

BACKGROUND: The World Health Organization (WHO) recommends the administration of intramuscular antenatal corticosteroids to women at risk of preterm birth to prevent preterm-associated neonatal mortality and morbidity. Poor quality medicines are a major problem for health services in low- and middle-income countries (LMICs), however the quality of antenatal corticosteroids is not well understood. We aimed to conduct a systematic review of available studies describing the quality of recommended injectable antenatal corticosteroids (dexamethasone or betamethasone) in LMICs. METHODS: Structured search strategy was applied to six databases (MEDLINE, EMBASE, CINAHL, International Pharmaceutical Abstracts, Global Index Medicus, WHO Medicines Quality Database), without year or language restrictions. Any primary study reporting any medicine quality parameter (Active Pharmacological Ingredient, pH and sterility) for injectable dexamethasone or betamethasone was eligible. Two authors independently screened studies for eligibility, extracted data on included studies and applied Medicine Quality Assessment Reporting Guidelines tool to assess study quality. Results were reported narratively, stratified by country of manufacture, organisation type and level of care. RESULTS: In total, 15,547 citations were screened with two eligible studies identified that focussed on dexamethasone quality (no studies of betamethasone were identified). One study included 19 samples from 9 LMICs, and the other included "less than 100 samples" from India. The prevalence of failed dexamethasone samples ranged from 3.14% to 32.2% due to inadequate Active Pharmacological Ingredient. A higher prevalence of failed dexamethasone samples were seen at the point of care and the public sector. CONCLUSIONS: Poor quality maternal and newborn health medicines can endanger women and newborns. Though available evidence on antenatal corticosteroids quality in LMICs is limited, results suggested poor quality dexamethasone may be prevalent in some countries. More primary studies are required to confirm these findings and guide policymakers on procurement of good-quality maternal and newborn health medicines

Male Flat Jockeys Do Not Display Deteriorations in Bone Density or Resting Metabolic Rate in Accordance With Race Riding Experience: Implications for RED-S.

Despite consistent reports of poor bone health in male jockeys, it is not yet known if this is a consequence of low energy availability or lack of an osteogenic stimulus. Given the rationale that low energy availability is a contributing factor in low bone health, we tested the hypothesis that both hip and lumbar bone mineral density (BMD) should progressively worsen in accordance with the years of riding. In a cross-sectional design, male apprentice (n=17) and senior (n=14) jockeys (matched for body mass and fat free mass) were assessed for hip and lumbar spine BMD as well as both measured and predicted resting metabolic rate (RMR). Despite differences (P0.05) in hip (-0.9 ± 1.1 v -0.8 ± 0.7) and lumbar Z-scores (-1.3 ± 1.4 v -1.5 ± 1) or measured RMR (1459 ± 160 v 1500 ± 165 kcal.d-1) between apprentices and senior jockeys, respectively. Additionally, years of race riding did not demonstrate any significant correlations (P>0.05) with either hip or lumbar spine BMD. Measured RMR was also not different (P>0.05) from predicted RMR in either apprentice (1520 ± 44 kcal.d-1) or senior jockeys (1505 ± 70 kcal.d-1). When considered with previously published data examining under-reporting of energy intake and direct assessments of energy expenditure, we suggest that low BMD in jockeys is not due to low energy availability per se, but rather, the lack of an osteogenic stimulus associated with riding

Cleaning of a model food soil from horizontal plates by a moving vertical water jet

The removal of layers of a model food soil (dried Xanthan gum containing fluorescent ZnS particles) by a vertical water jet impinging normally on to the plate, generated by a solid stream nozzle which moves across the plate was reported by Köhler et al. (2014). Their experiments investigated nozzle pressures from 0.5-2.0 barg; nozzle diameters from 0.84-2.66 mm, nozzle-layer separation of 20 mm, and nozzle traverse speeds of 2.1-126 mm s-1. The flow parameters and separation are smaller than those typical of industrial jet cleaning operations. The model developed by Wilson et al. (2014; Chem. Eng. Sci., 109, 183–196) for cleaning of similar layers by a stationary impinging jet was modified to describe the case of moving nozzle. This new model predicted the trends observed in the experiments, and analysis of the data yielded a similar cleaning rate constant to that obtained previously for cleaning of similar layers by stationary jets. The model predicted a non-circular cleaning front which matched that extracted from new experiments in which the flow was interrupted in order to capture this feature. The model allowed the cleaning performance indicators suggested by Köhler et al. (2014) to be expressed quantitatively: these indicated that higher nozzle traverse speeds give increased cleaning time, energy and liquid consumption performance.The work at TU Dresden was funded by the European Union and the Free State of Saxony as part of project SAB 080951793. A short vacation study grant for LC from Fitzwilliam College, Cambridge, is gratefully acknowledged.This is the accepted manuscript. The final version is available from Elsevier at http://www.sciencedirect.com/science/article/pii/S000925091400630

Similarity solutions for unsteady shear-stress-driven flow of Newtonian and power-law fluids : slender rivulets and dry patches

Unsteady flow of a thin film of a Newtonian fluid or a non-Newtonian power-law fluid with power-law index N driven by a constant shear stress applied at the free surface, on a plane inclined at an angle α to the horizontal, is considered. Unsteady similarity solutions representing flow of slender rivulets and flow around slender dry patches are obtained. Specifically, solutions are obtained for converging sessile rivulets (0 < α < π/2) and converging dry patches in a pendent film (π/2 < α < π), as well as for diverging pendent rivulets and diverging dry patches in a sessile film. These solutions predict that at any time t, the rivulet and dry patch widen or narrow according to |x|3/2, and the film thickens or thins according to |x|, where x denotes distance down the plane, and that at any station x, the rivulet and dry patch widen or narrow like |t|−1, and the film thickens or thins like |t|−1, independent of N

Spin Calogero Particles and Bispectral Solutions of the Matrix KP Hierarchy

Pairs of $n\times n$ matrices whose commutator differ from the identity by a matrix of rank $r$ are used to construct bispectral differential operators with $r\times r$ matrix coefficients satisfying the Lax equations of the Matrix KP hierarchy. Moreover, the bispectral involution on these operators has dynamical significance for the spin Calogero particles system whose phase space such pairs represent. In the case $r=1$, this reproduces well-known results of Wilson and others from the 1990's relating (spinless) Calogero-Moser systems to the bispectrality of (scalar) differential operators. This new class of pairs $(L, \Lambda)$ of bispectral matrix differential operators is different than those previously studied in that $L$ acts from the left, but $\Lambda$ from the right on a common $r\times r$ eigenmatrix.Comment: 16 page

GB Apprentice Jockeys Do Not Have the Body Composition to Make Current Minimum Race Weights: Is It Time to Change the Weights or Change the Jockeys?

Flat jockeys in Great Britain (GB) are classified as apprentices if they are aged less than 26 years and/or have ridden less than 95 winners. To gain experience, apprentices are allocated a weight allowance of up to 7 lb (3.2 kg). Given that there is no off-season in GB flat horseracing, jockeys are required to maintain their racing weight all year round. In light of recent work determining that current apprentices are considerably heavier than previous generations and that smaller increases have been made in the minimum weight, the aim of this study was to assess if the minimum weight in GB was achievable. To make the minimum weight (50.8 kg) with the maximal weight allowance requires a body mass of ∼46.6 kg while maintaining a fat mass >2.5 kg (the lowest fat mass previously reported in weight-restricted males). Thirty-two male apprentice jockeys were assessed for body composition using dual-energy X-ray absorptiometry. The mean (SD) total mass and fat mass were 56 (2.9) kg and 7.2 (1.8) kg, respectively. Given that the lowest theoretical body mass for this group was 51.2 (2.3) kg, only one of 32 jockeys was deemed feasible to achieve the minimum weight with their current weight allowance and maintaining fat mass >2.5 kg. Furthermore, urine osmolality of 780 (260) mOsmol/L was seen, with 22 (out of 32) jockeys classed as dehydrated (>700 mOsmols/L), indicating that body mass would be higher when euhydrated. Additionally, we observed that within new apprentice jockeys licensed during this study (N = 41), only one jockey was able to achieve the minimum weight. To facilitate the goal of achieving race weight with minimal disruptions to well-being, the authors’ data suggest that the minimum weight for GB apprentices should be raised

Conscious monitoring and control (reinvestment) in surgical performance under pressure.

Research on intraoperative stressors has focused on external factors without considering individual differences in the ability to cope with stress. One individual difference that is implicated in adverse effects of stress on performance is "reinvestment," the propensity for conscious monitoring and control of movements. The aim of this study was to examine the impact of reinvestment on laparoscopic performance under time pressure