Stimulation of intermolecular ligation with E. coli DNA ligase by high concentrations of monovalent cations in polyethylene glycol solutions.
In the presence of high concentrations of the nonspecific polymer polyethylene glycol (PEG), intermolecular cohesive-end ligation with the DNA ligase from Escherichia coli was stimulated by high salt concentrations: 200 mM NaCl or 300 mM KCl in 10% (w/v) PEG 6000 solutions, and 100-200 mM NaCl or 150-300 mM KCl in 15% PEG 6000 solutions. Intermolecular blunt-end ligation with this ligase was also stimulated at 100-150 mM NaCl or 150-250 mM KCl in 15% PEG 6000 solutions. The extent of such intermolecular ligation increased and the salt concentrations at which ligation was stimulated extended to lower concentrations when we raised the temperature from 10 to 37 degrees C.


Diversity in approach to teaching and assessing ethics education for medical undergraduates: A scoping review.
There are diverse methods to teach medical ethics, and there is no single accepted approach towards its learning and assessment. The authors aim to explore the various strategies practised to teach undergraduate medical students the fundamentals of medical ethics and their evaluation. The authors reviewed the articles published from January 2014 to September 2019. The authors searched PubMed for the relevant publications and extracted the information using a data extraction sheet. Twenty-nine articles were included for the review, which fulfilled the inclusion criteria. Case-based discussions were a widely accepted strategy to learn ethics. The studies highlighted a mixed teaching approach using multiple teaching tools. A qualitative approach was preferred for the assessment through reflections, simulated patient interactions, and development of portfolios. However, there are gaps in the existing literature on the assessment strategies for ethics education. Heterogeneity still exists in the planning of the curricula, teaching, and assessment methods. These curricula suit the cultural and religious set up of that particular country. Case-based discussion is a popular teaching strategy, and there exist numerous innovative and cost-effective active teaching strategies. There is a need for studies that are more rigorous to address the evaluation of the ethics curricula. This review would help educators to choose their preferred approach based on their teaching environment.


Tree growth influenced by warming winter climate and summer moisture availability in northern temperate forests.
The role of future forests in global biogeochemical cycles will depend on how different tree species respond to climate. Interpreting the response of forest growth to climate change requires an understanding of the temporal and spatial patterns of seasonal climatic influences on the growth of common tree species. We constructed a new network of 310 tree-ring width chronologies from three common tree species (Quercus robur, Pinus sylvestris and Fagus sylvatica) collected for different ecological, management and climate purposes in the south Baltic Sea region at the border of three bioclimatic zones (temperate continental, oceanic, southern boreal). The major climate factors (temperature, precipitation, drought) affecting tree growth at monthly and seasonal scales were identified. Our analysis documents that 20th century Scots pine and deciduous species growth is generally controlled by different climate parameters, and that summer moisture availability is increasingly important for the growth of deciduous species examined. We report changes in the influence of winter climate variables over the last decades, where a decreasing influence of late winter temperature on deciduous tree growth and an increasing influence of winter temperature on Scots pine growth was found. By comparing climate-growth responses for the 1943-1972 and 1973-2002 periods and characterizing site-level growth response stability, a descriptive application of spatial segregation analysis distinguished sites with stable responses to dominant climate parameters (northeast of the study region), and sites that collectively showed unstable responses to winter climate (southeast of the study region). The findings presented here highlight the temporally unstable and nonuniform responses of tree growth to climate variability, and that there are geographical coherent regions where these changes are similar. Considering continued climate change in the future, our results provide important regional perspectives on recent broad-scale climate-growth relationships for trees across the temperate to boreal forest transition around the south Baltic Sea.


Ethical aspects of genetic disease and genetic counselling.
With the reduction in diseases due to nutritional deficiencies and infection, disorders which are wholly or partly genetic are becoming relatively more important in all branches of modern medicine. Genetic counselling has developed in recent years from just explaining to an individual or a couple the risk of them producing a handicapped child, to the possibility in many cases of better diagnosis and active intervention to reduce the risks. At the same time antenatal screening programmes have been introduced to detect women who may be carrying a fetus with a severe handicapping anomaly. The ethical aspects of these advances are considered in this article. A practical approach to the resolution of any dilemmas is proposed which concentrates on the duties incumbent on doctors and other health care workers involved with patients who have or may carry genetic disorders.


Enabling relational leadership in primary healthcare settings: lessons from the DIALHS collaboration.
Strong management and leadership competencies have been identified as critical in enhancing health system performance. While the need for strong health system leadership has been raised, an important undertaking for health policy and systems researchers is to generate lessons about how to support leadership development (LD), particularly within the crisis-prone, resource poor contexts that are characteristic of Low- and Middle-Income health systems. As part of the broader DIALHS (District Innovation and Action Learning for Health Systems Development) collaboration, this article reflects on 5 years of action learning and engagement around leadership and LD within primary healthcare (PHC) services. Working in one sub-district in Cape Town, we co-created LD processes with managers from nine PHC facilities and with the six members of the sub-district management team. Within this article, we seek to provide insights into how leadership is currently practiced and to highlight lessons about whether and how our approach to LD enabled a strengthening of leadership within this setting. Findings suggest that the sub-district is located within a hierarchical governance context, with performance monitored through the use of multiple accountability mechanisms including standard operating procedures, facility audits and target setting processes. This context presents an important constraint to the development of a more distributed, relational leadership. While our data suggest that gains in leadership were emerging, our experience is of a system struggling to shift from a hierarchical to a more relational understanding of how to enable improvements in performance, and to implement these changes in practice.


Differences in 24-hour urine composition between black and white women.
Black women are less likely to develop kidney stones and have greater bone mass than white women. However, little is known about racial differences in urine composition. Urine pH, volume, and 24-h urinary excretion of calcium, citrate, oxalate, uric acid, sodium, potassium, magnesium, phosphate, sulfate, and creatinine of 146 black women were compared with 330 white women in the Nurses' Health Study. All participants were postmenopausal non-stone formers. ANOVA was used to compare mean urinary values. Linear regression models were adjusted for age, body mass index, dietary intake, and urinary factors. On average, black women excreted 65 mg less urinary calcium (P < 0.001), 4 mg more oxalate (P < 0.001), 9 mEq less potassium (P < 0.001), 11 mg less magnesium (P = 0.003), 120 mg less phosphate (P < 0.001), and 3 mmol less sulfate (P < 0.001) per day than did white women. The urine pH of black women was 0.11 units higher (P = 0.03) and urine volume was 0.24 L less (P = 0.001). The urinary relative supersaturations of calcium oxalate (P = 0.03) and brushite (P = 0.002) were lower in black women. No other significant differences were observed. Differences in urinary calcium and pH persisted after multivariate adjustment and after exclusion of participants who were taking thiazide diuretics or those with diabetes. In conclusion, black women excrete less urinary calcium and have a higher urinary pH than do white women. These differences are not explained by differences in age, body mass index, or diet and may account for the lower incidence of both nephrolithiasis and osteoporosis in black women.


A Computational Study on the LCST Phase Transition of a POEGMA Type Thermoresponsive Block Copolymer: Effect of Water Ordering and Individual Behavior of Blocks.
The atomistic origin underlying the lower critical solution temperature (LCST) behavior of thermoresponsive copolymers in water is still elusive. Here, we report all-atom molecular dynamics simulations of block copolymers of 2-(2-methoxyethoxy)ethyl methacrylate (MEO2MA) and oligo(ethylene glycol) methyl ether methacrylate (OEGMA300) in water at various block ratios and at temperatures below and above the LCST values of each homopolymer block. Our single chain simulations showed that hydration water molecules accumulate particularly near the side chain carbon atoms by forming ordered cage-like structures via extensive hydrogen bonding between them. These water cage formations surround the entire surface of PMEO2MA- b-POEGMA300 and enable the copolymer to remain in water below the LCST. As the temperature increases, each block exhibits a separate coil-to-globule transition above its own LCST. A detailed analysis of the interactions between polymer-water and water-water revealed that this phase transition is mainly driven by the reduced local water ordering by the disruption of the water cages when the temperature is increased above the LCST. We found that the transition occurs differently in the copolymer than the POEGMA homopolymers due to the interaction of the blocks, especially around the joint of the blocks. Accordingly, the phase transition of a block acts as an additional disruptive effect on the other block's water cage structure, which reduces the LCST values of PMEO2MA and POEGMA300 in the copolymer, compared to their individual single chain homopolymers.


Cell proliferation is a key determinant of the outcome of FOXO3a activation.
The FOXO family of forkhead transcription factors have a pivotal role in determining cell fate in response to oxidative stress. FOXO activity can either promote cell survival or induce cell death. Increased FOXO-mediated cell death has been implicated in the pathogenesis of degenerative diseases affecting musculoskeletal tissues. The aim of this study was to determine the conditions under which one member of the FOXO family, FOXO3a, promotes cell survival as opposed to cell death. Treatment of primary human tenocytes with 1 pM hydrogen peroxide for 18 h resulted in increased protein levels of FOXO3a. In peroxide-treated cells cultured in low serum media, FOXO3a inhibited cell proliferation and protected against apoptosis. However in peroxide treated cells cultured in high serum media, cell proliferation was unchanged but level of apoptosis significantly increased. Similarly, in tenocytes transduced to over-express FOXO3a, cell proliferation was inhibited and level of apoptosis unchanged in cells cultured in low serum. However there was a robust increase in cell death in FOXO3a-expressing cells cultured in high serum. Inhibition of cell proliferation in either peroxide-treated or FOXO3a-expressing cells cultured in high serum protected against apoptosis induction. Conversely, addition of a Chk2 inhibitor to peroxide-treated or FOXO3a-expressing cells overrode the inhibitory effect of FOXO3a on cell proliferation and led to increased apoptosis in cells cultured in low serum. This study demonstrates that proliferating cells may be particularly susceptible to the apoptosis-inducing actions of FOXO3a. Inhibition of cell proliferation by FOXO3a may be a critical event in allowing the pro-survival rather than the pro-apoptotic activity of FOXO3a to prevail.


Myogenic response of isolated pressurized rabbit ear artery is independent of endothelium.
Central ear artery segments, removed from urethan-anesthetized rabbits, were used to assess whether distention activation was dependent on intact and functional endothelium (ENDO). Changes in external arterial diameter were measured with light-dependent diode array mounted above the vessel. After constriction with norepinephrine, slow increases in pressure from 60 to 80 or 100 mmHg performed over 120 s (pressure ramps) failed to initiate distention, but initial diameter was maintained. Rapidly applied (500 ms) pressure increases (pressure jumps) across same pressure ranges gave rise to initial distention and a myogenic response resulting in a return to almost initial diameter while new pressure was maintained for 120 s. Myogenic activity was measured from extent of recovery of vessel to its initial diameter during maintenance of pressure jumps or ramps. Rapid jumps and slow ramps were performed in presence of an intact ENDO and again after ENDO was removed by passage of intraluminal gas. With pressure jumps 60 to 80 mmHg, degree of recovery in ENDO-denuded vessels was 91.9% compared with 89.5% in ENDO intact vessels. For pressure jumps 60 to 100 mmHg, extent of recovery was 87.4 and 89.6%. During application of pressure ramps of 20 or 40 mmHg, vessel diameter did not increase by greater than 5%. There are no significant differences in these data, and we conclude that myogenic response in rabbit ear artery is mediated independently of endothelial-derived factors, irrespective of whether myogenic activation is induced by pressure jumps or ramps.


Genome-Wide Identification and Characterization of Salvia miltiorrhiza Laccases Reveal Potential Targets for Salvianolic Acid B Biosynthesis.
Laccases are widely distributed in plant kingdom catalyzing the polymerization of lignin monolignols. Rosmarinic acid (RA) has a lignin monolignol-like structure and is converted into salvianolic acid B (SAB), which is a representatively effective hydrophilic compound of a well-known medicinal plant Salvia miltiorrhiza and also the final compound of phenolic acids metabolism pathway in the plant. But the roles of laccases in the biosynthesis of SAB are poorly understood. This work systematically characterizes S. miltiorrhiza laccase (SmLAC) gene family and identifies the SAB-specific candidates. Totally, 29 laccase candidates (SmLAC1-SmLAC29) are found to contain three signature Cu-oxidase domains. They present relatively low sequence identity and diverse intron-exon patterns. The phylogenetic clustering of laccases from S. miltiorrhiza and other ten plants indicates that the 29 SmLACs can be divided into seven groups, revealing potential distinct functions. Existence of diverse cis regulatory elements in the SmLACs promoters suggests putative interactions with transcription factors. Seven SmLACs are found to be potential targets of miR397. Putative glycosylation sites and phosphorylation sites are identified in SmLAC amino acid sequences. Moreover, the expression profile of SmLACs in different organs and tissues deciphers that 5 SmLACs (SmLAC7/8/20/27/28) are expressed preferentially in roots, adding the evidence that they may be involved in the phenylpropanoid metabolic pathway. Besides, silencing of SmLAC7, SmLAC20 and SmLAC28, and overexpression of SmLAC7 and SmLAC20 in the hairy roots of S. miltiorrhiza result in diversification of SAB, signifying that SmLAC7 and SmLAC20 take roles in SAB biosynthesis. The results of this study lay a foundation for further elucidation of laccase functions in S. miltiorrhiza, and add to the knowledge for SAB biosynthesis in S. miltiorrhiza.


Avoiding Accidents at the Champagne Reception.
Using a lifting and balancing task, we contrasted two alternative views of planning joint actions: one postulating that joint action involves distinct predictions for self and other, the other postulating that joint action involves coordinated plans between the coactors and reuse of bimanual models. We compared compensatory movements required to keep a tray balanced when 2 participants lifted glasses from each other's trays at the same time (simultaneous joint action) and when they took turns lifting (sequential joint action). Compared with sequential joint action, simultaneous joint action made it easier to keep the tray balanced. Thus, in keeping with the view that bimanual models are reused for joint action, predicting the timing of their own lifting action helped participants compensate for another person's lifting action. These results raise the possibility that simultaneous joint actions do not necessarily require distinguishing between one's own and the coactor's contributions to the action plan and may afford an agent-neutral stance.


Involvement of Akt-1 and mTOR in sensitivity of breast cancer to targeted therapy.
Elucidating the response of breast cancer cells to chemotherapeutic and hormonal based drugs is clearly important as these are frequently used therapeutic approaches. A signaling pathway often involved in chemo- and hormonal-resistance is the Ras/PI3K/PTEN/Akt/mTOR cascades. In the studies presented in this report, we have examined the effects of constitutive activation of Akt on the sensitivity of MCF-7 breast cancer cells to chemotherapeutic- and hormonal-based drugs as well as mTOR inhibitors. MCF-7 cells which expressed a constitutively-activated Akt-1 gene [∆Akt-1(CA)] were more resistant to doxorubicin, etoposide and 4-OH-tamoxifen (4HT) than cells lacking ∆Akt-1(CA). Cells which expressed ∆Akt-1(CA) were hypersensitive to the mTOR inhibitor rapamycin. Furthermore, rapamycin lowered the IC50s for doxorubicin, etoposide and 4HT in the cells which expressed ∆Akt-1(CA), demonstrating a potential improved method for treating certain breast cancers which have deregulated PI3K/PTEN/Akt/mTOR signaling. Understanding how breast cancers respond to chemo- and hormonal-based therapies and the mechanisms by which they can become drug resistant may enhance our ability to treat breast cancer. These results also document the potential importance of knowledge of the mutations present in certain cancers which may permit more effective therapies.


The assessment of symptoms and functional limitations in low back pain patients: validity and reliability of a new questionnaire.
Many of the existing low back pain (LBP) questionnaires of function and symptoms have a content of different domains of disability presented as a single sum score, making it difficult to derive changes within a specific domain. The present study describes the development of a clinically derived back-specific questionnaire incorporating both a functional limitation and a symptom scale, with a further subdivision of the symptom scale in separate indices for severity and temporal aspects. The aims of the study were to assess the overall reliability and validity of the new questionnaire, named the Profile Fitness Mapping questionnaire (PFM). A total of 193 chronic LBP patients answered the PFM together with five validated criterion questionnaires. For the internal consistency of the questionnaires, the three indices of the PFM had the highest Cronbach's alpha (0.90-0.95) and all items had item-total correlations above 0.2. The correlation coefficients between the PFM and the back-specific criterion questionnaires ranged between 0.61 and 0.83, indicating good concurrent criterion validity. The best discriminative ability between patients with different pain severities was demonstrated by the functional limitation scale of the PFM. Well centered score distribution with no patient's score at the floor or the ceiling level indicates that the PFM has the potential to detect the improvement or worsening of symptoms and functional limitations in chronic LBP patients. Classification according to the International Classification of Functioning, Disability and health (ICF) of WHO revealed a high degree of homogeneous item content of the symptom scale to the domain of impairments, and of the functional limitation scale to the domain of activity limitations. The present study suggests that the PFM has a high internal consistency and is a valid indicator of symptoms and functional limitations of LBP patients. It offers the combination of a composite total score and the possibility of evaluations within specific domains of disability. Complementary evaluation of test-retest reliability and responsiveness to change is warranted.


Floor slat openings impact ammonia and greenhouse gas emissions associated with group-housed gestating sows.
According to EU legislation, group-housed gestating sows must have a minimum of 2.25 m2 floor area per sow with at least 1.3 m2 of continuous solid floor of which a maximum of 15% is reserved for drainage openings. The aim of the experiment was to quantify the impact of different drainage openings on ammonia and greenhouse gas emissions. Three successive batches of 10 gestating sows were used. Each batch was divided into two groups kept separately in two identical rooms with similar volume and surface. The solid part of the floor presented 15% drainage openings in the first room and 2.5% in the second room. The gas emissions (ammonia (NH3), methane (CH4), nitrous oxide (N2O), carbon dioxide (CO2) and water vapour (H2O)) were measured three times during 6 consecutive days. Gaseous emissions were significantly lower with 15% drainage openings with reductions of 19% for NH3 (12.77 v. 15.83 g/day per sow), 15% for CH4 (10.15 v. 11.91 g/day per sow), 10% for N2O (0.47 v. 0.52 g/day per sow), 9% for CO2 (2.41 v. 2.66 kg/day per sow) and 13% for H2O (3.25 v. 3.75 kg/day per sow). This trial showed the advantage, in an environmental point of view, to use 15% drainage openings on the solid part of partly slatted floors in pens for group-housed gestating sows.


The changing pattern of referral in acute kidney injury.
Acute kidney injury (AKI) is not only managed by nephrologists, but also by several other subspecialists. The referral rate to nephrologists and the factors influencing it are unknown. To determine the referral rate, factors affecting referral and outcomes across the spectrum of AKI in a population based study. We identified all patients with serum creatinine concentrations ≥150 µmol/l (male) or ≥130 µmol/l (female) over a 6-month period. AKI was defined according to the RIFLE classification (risk, injury, failure, loss, end stage renal disease [ESRD]). Clinical information and outcomes were obtained from each patient's case records. A total of 562 patients were identified as having AKI (incidence 2147 per million population/year [pmp/y]). One hundred and sixty-four patients (29%) were referred to nephrologists-referral rate 627 pmp/y. Forty-nine percent of patients whose serum creatinine rose to >300 µmol/l were referred compared with 22% in our previous study of 1997. Forty-eight patients required renal replacement therapy-incidence 184 pmp/y in comparison to 50 pmp/y in our previous study of 1997. Patients had higher odds of referral if they were male, of younger age and were in the F category of the RIFLE classification. Patients had lower odds of referral if they had multiple co-morbid conditions or if they were managed in a hospital without a nephrology service. There has been a significant rise in the referral rate of patients with AKI to nephrologists but even during our period of study only one-third of such patients were being referred. With rising incidence and increased awareness, the referral rate will certainly rise putting a significant burden on the nephrology services.


Pregnancy outcomes in healthy nulliparas who developed hypertension. Calcium for Preeclampsia Prevention Study Group.
To determine maternal and perinatal outcomes in nulliparas with pregnancy-associated hypertension or preeclampsia. We conducted (and reported elsewhere) a randomized, double-masked, placebo-controlled trial calcium supplementation of 4589 healthy nulliparas assigned at 13-21 weeks' gestation. This well-defined and characterized data set provided an opportunity to detail more precisely adverse maternal, fetal, and newborn outcomes in women who developed hypertension among a prospective series of healthy nulliparas. Of 4302 women observed to or beyond 20 weeks' gestation, 1073 (24.9%) developed mild or severe pregnancy-associated hypertension or preeclampsia. One hundred sixteen women of the 1073 with hypertension (10.8%) and 336 of the 3229 without hypertension (10.4%) were delivered before 37 weeks' gestation. Fetal and neonatal mortality were similar in those groups; however, selected maternal and newborn morbidities were significantly greater in women with hypertension. Significantly increased maternal morbidities included increased cesarean deliveries, abruptio placentae, and acute renal dysfunction; and significantly increased perinatal morbidities included respiratory distress syndrome, ventilatory support, and fetal growth restriction. Adverse outcomes were highest in women with severe pregnancy-associated hypertension or preeclampsia. Hypertension, especially severe hypertension, was associated with an appreciable increase in important maternal and perinatal morbidity but not perinatal mortality.


Nephritogenicity of antibodies to proteoglycans of the glomerular basement membrane--I.
We investigated nephritogenic potential of antibodies to heparan sulfate-proteoglycan of glomerular basement membrane. Glomeruli were isolated, basement membranes were prepared, proteoglycans extracted, and purified core protein was obtained. We immunized rabbits with the core protein, IgG fraction prepared from the antisera and specificity of the antibody determined. A single immunoprecipitin line in agar diffusion plate and a single band (approximately 18,000 mol wt) on the immunoblot autoradiograms were visualized. The antibody showed precise reactivity with the glomerular basement membranes. The clearance studies indicated that approximately 75% of the radioiodinated antibody disappeared from circulation within 1 h and 1-2% bound to the kidney. For nephritogenicity experiments, the antibody was intravenously administered into rats and we examined their kidneys at 1 h to 24 d later. A linear immunofluorescence of glomerular basement membranes was observed with rabbit IgG at all times while that of C3 until the 10th day. Early morphologic changes included glomerular infiltration of polymorphonuclear leukocytes with focal exfoliation of endothelium. The leukocytic infiltration subsided by the third day and was followed by progressive thickening of basement membranes, focal mesangial cell proliferation, increase in mesangial matrix, and accumulation of monocytes. Focal knob-like thickening of glomerular basement membrane was observed from the 15th day onward. Regularly-spaced electrondense deposits were seen in the lamina rara interna and externa of glomerular basement membranes and persisted throughout the investigatory period. No significant proteinuria was observed at any stage of the experiment. These findings suggest that the antibodies to the basement membrane heparan sulfate-proteoglycan are nephrotoxic but possess weak nephritogenic potential.


Nut consumption in relation to all-cause and cause-specific mortality: a meta-analysis 18 prospective studies.
Several previous meta-analyses show a consistent inverse association between nut consumption and all-cause mortality, but the associations with cause-specific mortality remain uncertain. A recent meta-analysis on nut consumption and multiple health outcomes combined incidence and mortality outcomes across most of the analyses, which may have introduced heterogeneity across studies. We conducted an updated meta-analysis to evaluate the nut-mortality association. We searched PubMed and EMBASE and we contacted authors for additional data. The final analyses included 18 prospective studies. The random-effects summary RRs for high compared with low nut consumption were 0.81 (95% CI: 0.78-0.84) for all-cause mortality (18 studies with 81 034 deaths), 0.75 (95% CI: 0.71-0.79) for CVD mortality (17 studies with 20 381 deaths), 0.73 (95% CI: 0.67-0.80) for CHD mortality (14 studies with 10 438 deaths), 0.82 (95% CI: 0.73-0.91) for stroke mortality (13 studies with 4850 deaths) and 0.87 (95% CI: 0.80-0.93) for cancer mortality (11 studies 21 353 deaths). These results were broadly consistent within subgroups according to various study and population characteristics and within sensitivity analyses that took into account potential confounders. Peanut (5 studies) and tree nut (3 studies) consumption were similarly associated with mortality risks. Dose-response analyses suggested evidence for nonlinear associations between nut consumption and mortality (P-nonlinearity <0.001 for all outcomes except cancer mortality), with mortality risk levelling off at the consumption of about 3 servings per week (12 g d-1). Our findings suggest that nut consumption is associated with reduced all-cause and cause-specific mortality, with the strongest reduction for CHD mortality. Both tree nuts and peanuts may lower mortality and most of the survival benefits may be achieved at a relative low level of nut consumption.


Colour Constancy for Image of Non-Uniformly Lit Scenes.
Digital camera sensors are designed to record all incident light from a captured scene, but they are unable to distinguish between the colour of the light source and the true colour of objects. The resulting captured image exhibits a colour cast toward the colour of light source. This paper presents a colour constancy algorithm for images of scenes lit by non-uniform light sources. The proposed algorithm uses a histogram-based algorithm to determine the number of colour regions. It then applies the K-means++ algorithm on the input image, dividing the image into its segments. The proposed algorithm computes the Normalized Average Absolute Difference (NAAD) for each segment and uses it as a measure to determine if the segment has sufficient colour variations. The initial colour constancy adjustment factors for each segment with sufficient colour variation is calculated. The Colour Constancy Adjustment Weighting Factors (CCAWF) for each pixel of the image are determined by fusing the CCAWFs of the segments, weighted by their normalized Euclidian distance of the pixel from the center of the segments. Results show that the proposed method outperforms the statistical techniques and its images exhibit significantly higher subjective quality to those of the learning-based methods. In addition, the execution time of the proposed algorithm is comparable to statistical-based techniques and is much lower than those of the state-of-the-art learning-based methods.


Thyroid function in pregnancy: what is normal?
Gestational thyroid dysfunction is common and associated with maternal and child morbidity and mortality. During pregnancy, profound changes in thyroid physiology occur, resulting in different thyroid-stimulating hormone (TSH) and free thyroxine (FT4) reference intervals compared to the nonpregnant state. Therefore, international guidelines recommend calculating trimester- and assay-specific reference intervals per center. If these reference intervals are unavailable, TSH reference intervals of 0.1-2.5 mU/L for the first trimester and 0.2-3.0 mU/L for the second trimester are recommended. In daily practice, most institutions do not calculate institution-specific reference intervals but rely on these fixed reference intervals for the diagnosis and treatment of thyroid disorders during pregnancy. However, the calculated reference intervals for several additional pregnancy cohorts have been published in the last few years and show substantial variation. We provide a detailed overview of the available studies on thyroid function reference intervals during pregnancy, different factors that contribute to these reference intervals, and the maternal and child complications associated with only minor variations in thyroid function. There are large differences in thyroid function reference intervals between different populations of pregnant women. These differences can be explained by variations in assays as well as population-specific factors, such as ethnicity and body mass index. The importance of using correct reference intervals is underlined by the fact that even small subclinical variations in thyroid function have been associated with detrimental pregnancy outcomes, including low birth weight and pregnancy loss. It is therefore crucial that institutions do not rely on fixed universal cutoff concentrations, but calculate their own pregnancy-specific reference intervals.


Competent statistical programmer: Need of business process outsourcing industry.
Over the last two decades Business Process Outsourcing (BPO) has evolved as much mature practice. India is looked as preferred destination for pharmaceutical outsourcing over a cost arbitrage. Among the biometrics outsourcing, statistical programming and analysis required very niche skill for service delivery. The demand and supply ratios are imbalance due to high churn out rate and less supply of competent programmer. Industry is moving from task delivery to ownership and accountability. The paradigm shift from an outsourcing to consulting is triggering the need for competent statistical programmer. Programmers should be trained in technical, analytical, problem solving, decision making and soft skill as the expectations from the customer are changing from task delivery to accountability of the project. This paper will highlight the common issue SAS programming service industry is facing and skills the programmers need to develop to cope up with these changes.


Identification of capric acid as a potent vasorelaxant of human basilar arteries.
To determine whether naturally occurring fatty acids, especially saturated ones, might act directly as vasodilators, segments of human basilar arteries and umbilical arteries were precontracted submaximally with prostaglandin F2 alpha and then exposed to different saturated fatty acids (C4 through C16) or unsaturated fatty acids (C14:1, C18:1, C18:2, and C18:3) at concentrations from 4 microM to 4 mM. The results showed caprate (C10) to be the most potent vasorelaxant and basilar arteries to be more responsive (EC50 = 63 microM) than umbilical arteries (EC50 = 780 microM). Caprate also inhibited contractions elicited by KCl, serotonin, and the thromboxane analogue U46619. The relaxation was independent of the endothelium, and potency was not related to the weak capacity of caprate to inhibit Ca(2+)-induced contractions of K(+)-depolarized basilar arteries. The pattern of potencies for the arteries differed, but among unsaturated fatty acids the monounsaturated (C14:1, C18:1) were more potent than the polyunsaturated (C18:2, C18:3). Comparing the potencies obtained with the concentrations reported for the free fatty acid content of arteries, brain, and plasma indicates that these lipids could influence vasomotion in health and disease.


Repression of apurinic/apyrimidinic endonuclease by p53-dependent apoptosis in hydronephrosis-induced rat kidney.
p53 plays a major role in apoptosis through activation of pro-apoptotic gene Bax. It also regulates apurinic/apyrimidinic endonuclease (APE) expression in the base excision repair pathway against oxidative DNA damages. This study investigated whether p53-dependent apoptosis is correlated with APE using an experimental rat model of hydronephrosis. Hydronephrosis was induced by partial ligation of the right ureter. Animals were sacrificed on scheduled time after unilateral ureteral obstruction and the expression of 8-OHdG, γ-H2AX, apoptotic proteins and APE was determined. The accumulated p53 activated Bax and caspase-3 7 days after hydronephrosis induction and the resulting high levels of p53-dependent apoptotic proteins and γ-H2AX tended to decrease APE. The intensities of 8-OHdG and caspase-3 immunolocalization significantly increased in obstructed kidneys than in sham-operated kidneys, although APE immunoreactivity increased after hydronephrosis induction. These results suggest that oxidative DNA damages in obstructed kidneys may trigger p53-dependent apoptosis through repression of APE.


A Systematic Review and Meta-analysis Comparing the Effectiveness and Adverse Effects of Different Systemic Treatments for Non-clear Cell Renal Cell Carcinoma.
While vascular endothelial growth factor-targeted therapy and mammalian target of rapamycin inhibition are effective strategies in treating clear cell renal cell carcinoma (ccRCC), the most effective therapeutic approach for patients with non-clear cell RCC (non-ccRCC) is unknown. To systematically review relevant literature comparing the oncological outcomes and adverse events of different systemic therapies for patients with metastatic non-ccRCC. Relevant databases including MEDLINE, Embase, and the Cochrane Library were searched up to March 24, 2016. Only comparative studies were included. Risk of bias and confounding assessments were performed. A meta-analysis was planned for and only performed if methodologically appropriate; otherwise, a narrative synthesis was undertaken. The literature search identified 812 potential titles and abstracts. Five randomized controlled trials, recruiting a total of 365 patients, were included. Three studies compared sunitinib against everolimus, one of which reported the results for non-ccRCC as a subgroup rather than as an entire randomized cohort. Individually, the studies showed a trend towards favoring sunitinib in terms of overall survival and progression-free survival (PFS; Everolimus versus Sunitinib in Patients with Metastatic Non-clear Cell Renal Cell Carcinoma hazard ratio [HR]: 1.41, 80% confidence interval [CI] 1.03-1.92 and 1.41, 95% CI: 0.88-2.27, Evaluation in Metastatic Non-clear Cell Renal Cell Carcinoma HR: 1.16, 95% CI: 0.67-2.01, Efficacy and Safety Comparison of RAD001 Versus Sunitinib in the First-line and Second-line Treatment of Patients with Metastatic Renal Cell Carcinoma HR: 1.5, 95% CI: 0.9-2.8), but this trend did not reach statistical significance in any study. Meta-analysis was performed on two studies which solely recruited patients with non-ccRCC reporting on PFS, the results of which were inconclusive (HR: 1.30, 95% CI: 0.91-1.86). Sunitinib was associated with more Grade 3-4 adverse events than everolimus, although this was not statistically significant. This systematic review and meta-analysis represent a robust summary of the evidence base for systemic treatment of metastatic non-ccRCC. The results show a trend towards favoring vascular endothelial growth factor-targeted therapy for PFS and overall survival compared with mammalian target of rapamycin inhibitors, although statistical significance was not reached. The relative benefits and harms of these treatments remain uncertain. Further research, either in the form of an individual patient data meta-analysis involving all relevant trials, or a randomized controlled trial with sufficient power to detect potential differences between treatments, is needed. We examined the literature to determine the most effective treatments for advanced kidney cancer patients whose tumors are not of the clear cell subtype. The results suggest that a drug called sunitinib might be more effective than everolimus, but the statistics supporting this statement are not yet entirely reliable. Further research is required to clarify this unmet medical need.


An embryological perspective on the early arthropod fossil record.
Our understanding of the early evolution of the arthropod body plan has recently improved significantly through advances in phylogeny and developmental biology and through new interpretations of the fossil record. However, there has been limited effort to synthesize data from these different sources. Bringing an embryological perspective into the fossil record is a useful way to integrate knowledge from different disciplines into a single coherent view of arthropod evolution. I have used current knowledge on the development of extant arthropods, together with published descriptions of fossils, to reconstruct the germband stages of a series of key taxa leading from the arthropod lower stem group to crown group taxa. These reconstruction highlight the main evolutionary transitions that have occurred during early arthropod evolution, provide new insights into the types of mechanisms that could have been active and suggest new questions and research directions. The reconstructions suggest several novel homology hypotheses - e.g. the lower stem group head shield and head capsules in the crown group are all hypothesized to derive from the embryonic head lobes. The homology of anterior segments in different groups is resolved consistently. The transition between "lower-stem" and "upper-stem" arthropods is highlighted as a major transition with a concentration of novelties and innovations, suggesting a gap in the fossil record. A close relationship between chelicerates and megacheirans is supported by the embryonic reconstructions, and I suggest that the depth of the mandibulate-chelicerate split should be reexamined.


The value of the modern vascular surgeon to the health care system: A report from the Society for Vascular Surgery Valuation Work Group.
Vascular surgeons provide an important service to the health care system. They are capable of treating a wide range of disease processes that affect both the venous and arterial systems. Their presence broadens the complexity and diversity of services that a health care system can offer both in the outpatient setting and in the inpatient setting. Because of their ability to control hemorrhage, they are critical to a safe operating room environment. The vascular surgery service line has a positive impact on hospital margin through both the direct vascular profit and loss and the indirect result of assisting other surgical and medical services in providing care. The financial benefits of a vascular service line will hold true for a wide range of alternative payment models, such as bundled payments or capitation. To fully leverage a modern vascular surgeon's skill set, significant investment is required from the health care system that is, however, associated with substantial return on the investment.


The exonuclease domain of Lassa virus nucleoprotein is involved in antigen-presenting-cell-mediated NK cell responses.
Lassa virus is an Old World Arenavirus which causes Lassa hemorrhagic fever in humans, mostly in West Africa. Lassa fever is an important public health problem, and a safe and effective vaccine is urgently needed. The infection causes immunosuppression, probably due to the absence of activation of antigen-presenting cells (dendritic cells and macrophages), low type I interferon (IFN) production, and deficient NK cell function. However, a recombinant Lassa virus carrying D389A and G392A substitutions in the nucleoprotein that abolish the exonuclease activity and IFN activation loses its inhibitory activity and induces strong type I IFN production by dendritic cells and macrophages. We show here that during infection by this mutant Lassa virus, antigen-presenting cells trigger efficient human NK cell responses in vitro, including production of IFN-γ and cytotoxicity. NK cell activation involves close contact with both antigen-presenting cells and soluble factors. We report that infected dendritic cells and macrophages express the NKG2D ligands major histocompatibility complex (MHC) class I-related chains A and B and that they may produce interleukin-12 (IL-12), IL-15, and IL-18, all involved in NK cell functions. NK cell degranulation is significantly increased in cocultures, suggesting that NK cells seem to kill infected dendritic cells and macrophages. This work confirms the inhibitory function of Lassa virus nucleoprotein. Importantly, we demonstrate for the first time that Lassa virus nucleoprotein is involved in the inhibition of antigen-presenting cell-mediated NK cell responses. The pathogenesis and immune responses induced by Lassa virus are poorly known. Recently, an exonuclease domain contained in the viral nucleoprotein has been shown to be able to inhibit the type I IFN response by avoiding the recognition of viral RNA by cell sensors. Here, we studied the responses of NK cells to dendritic cells and macrophages infected with a recombinant Lassa virus in which the exonuclease functions have been abolished and demonstrated that NK cells are strongly activated and presented effective functions. These results show that the strategy developed by Lassa virus to evade innate immunity is also effective on NK cells, explaining the weak NK cell activation observed with the wild-type virus. By providing a better understanding of the interactions between Lassa virus and the host immune system, these results are important for the field of arenavirus biology and may be useful for a vaccine approach against Lassa fever.


Giant Cornu Cutaneum Superimposed on Basal Cell Carcinoma.
Cornu cutaneum (CC) is a clinical term that describes the horn-like keratotic lesions extending vertically from the skin. Benign, premalignant or malignant lesions may be present at the base of CC. Seborrhoeic keratosis and squamous cell carcinoma (SCC) are the most commonly reported benign and malignant forms, respectively. Basal cell carcinoma (BCC) at the base is rare. Here, we report on an 85-year old female patient having multiple CC lesions, one being giant on her face and two of the lesions diagnosed with BCC at the base. This case is of significance due to the presence of giant and multiple CC and detection of BCC at the base of more than one lesion. This present case indicates the need for the treatment of possible malignant lesions underlying CC in the elderly by total surgical excision.


Yes-associated protein (YAP65) in relation to Smad7 expression in human pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is characterized by multiple alterations in the TGF-beta signaling pathway. Yes-associated protein (YAP65) interacts with Smad7 thereby influencing TGF-beta signaling. In the present study, the expression of YAP65 in PDAC was analyzed in order to elucidate the potential role of this molecule in the pathogenesis of pancreatic cancer. YAP65 mRNA expression levels in human pancreatic tissue samples and cell lines were analyzed by Northern blotting and quantitative RT-PCR. Immunohistochemistry was carried out to localize and quantify YAP65 expression in relation to Smad7 expression and Smad4 mutations. The effects of TGF-beta1 on Smad7 and YAP65 mRNA expression were analyzed by quantitative RT-PCR. Enhanced expression of YAP65 mRNA was identified by Northern blotting and quantitative RT-PCR in PDAC in comparison to the normal pancreas (2.5-fold increase) and to chronic pancreatitis (1.3-fold increase). In the normal pancreas, YAP65 was absent in acinar cells, large ducts and islet cells, but exhibited moderate to strong immunoreactivity in centroacinar cells and ductules. Tubular complexes in CP and CP-like lesions in PDAC also exhibited strong staining. In contrast, weak to moderate YAP65 immunoreactivity was present in the cancer cells. There was no correlation between YAP65 immunostaining and Smad7 staining or Smad4 mutations in the cancer samples. TGF-beta1 strongly induced Smad7 mRNA in Colo-357 and in Panc-1 cells, but only slightly induced YAP65 mRNA in Colo-357 cells. In conclusion, YAP65 is expressed mainly in centroacinar and small ductal cells in the normal pancreas. In PDAC, YAP65 is present in tubular complexes and to a lesser extent in cancer cells. Together with the known function of YAP65 in different growth and differentiation regulating pathways, it is suggested that this gene plays a role in the normal and diseased pancreas.


Ocean acidification through the lens of ecological theory.
Ocean acidification, chemical changes to the carbonate system of seawater, is emerging as a key environmental challenge accompanying global warming and other human-induced perturbations. Considerable research seeks to define the scope and character of potential outcomes from this phenomenon, but a crucial impediment persists. Ecological theory, despite its power and utility, has been only peripherally applied to the problem. Here we sketch in broad strokes several areas where fundamental principles of ecology have the capacity to generate insight into ocean acidification's consequences. We focus on conceptual models that, when considered in the context of acidification, yield explicit predictions regarding a spectrum of population- and community-level effects, from narrowing of species ranges and shifts in patterns of demographic connectivity, to modified consumer-resource relationships, to ascendance of weedy taxa and loss of species diversity. Although our coverage represents only a small fraction of the breadth of possible insights achievable from the application of theory, our hope is that this initial foray will spur expanded efforts to blend experiments with theoretical approaches. The result promises to be a deeper and more nuanced understanding of ocean acidification'and the ecological changes it portends.


Past SARS-CoV-2 infection protection against re-infection: a systematic review and meta-analysis.
Understanding the level and characteristics of protection from past SARS-CoV-2 infection against subsequent re-infection, symptomatic COVID-19 disease, and severe disease is essential for predicting future potential disease burden, for designing policies that restrict travel or access to venues where there is a high risk of transmission, and for informing choices about when to receive vaccine doses. We aimed to systematically synthesise studies to estimate protection from past infection by variant, and where data allow, by time since infection. In this systematic review and meta-analysis, we identified, reviewed, and extracted from the scientific literature retrospective and prospective cohort studies and test-negative case-control studies published from inception up to Sept 31, 2022, that estimated the reduction in risk of COVID-19 among individuals with a past SARS-CoV-2 infection in comparison to those without a previous infection. We meta-analysed the effectiveness of past infection by outcome (infection, symptomatic disease, and severe disease), variant, and time since infection. We ran a Bayesian meta-regression to estimate the pooled estimates of protection. Risk-of-bias assessment was evaluated using the National Institutes of Health quality-assessment tools. The systematic review was PRISMA compliant and was registered with PROSPERO (number CRD42022303850). We identified a total of 65 studies from 19 different countries. Our meta-analyses showed that protection from past infection and any symptomatic disease was high for ancestral, alpha, beta, and delta variants, but was substantially lower for the omicron BA.1 variant. Pooled effectiveness against re-infection by the omicron BA.1 variant was 45·3% (95% uncertainty interval [UI] 17·3-76·1) and 44·0% (26·5-65·0) against omicron BA.1 symptomatic disease. Mean pooled effectiveness was greater than 78% against severe disease (hospitalisation and death) for all variants, including omicron BA.1. Protection from re-infection from ancestral, alpha, and delta variants declined over time but remained at 78·6% (49·8-93·6) at 40 weeks. Protection against re-infection by the omicron BA.1 variant declined more rapidly and was estimated at 36·1% (24·4-51·3) at 40 weeks. On the other hand, protection against severe disease remained high for all variants, with 90·2% (69·7-97·5) for ancestral, alpha, and delta variants, and 88·9% (84·7-90·9) for omicron BA.1 at 40 weeks. Protection from past infection against re-infection from pre-omicron variants was very high and remained high even after 40 weeks. Protection was substantially lower for the omicron BA.1 variant and declined more rapidly over time than protection against previous variants. Protection from severe disease was high for all variants. The immunity conferred by past infection should be weighed alongside protection from vaccination when assessing future disease burden from COVID-19, providing guidance on when individuals should be vaccinated, and designing policies that mandate vaccination for workers or restrict access, on the basis of immune status, to settings where the risk of transmission is high, such as travel and high-occupancy indoor settings. Bill & Melinda Gates Foundation, J Stanton, T Gillespie, and J and E Nordstrom.


Leiomyoma of the gallbladder: a case report with review of the literature and discussion of the differential diagnosis.
Mesenchymal neoplasms of the gallbladder are rare, and most represent sarcomas of various histological types. To our knowledge, only a few patients with multiple Epstein-Barr virus (EBV)-associated smooth muscle tumours of the gallbladder in the setting of immunodeficiency have been reported in the English literature, but no single case of conventional leiomyoma has been well documented to date. A case of gallbladder leiomyoma in a healthy 34-year-old woman is described here. The tumour was found incidentally on a routine ultrasound examination and was removed by simple cholecystectomy. The patient is alive and well at last follow-up, 6 months after surgery. Histology and immunohistochemistry were consistent with a benign smooth muscle neoplasm that is very similar to conventional uterine leiomyoma. The tumour was negative for both EBV-encoded nuclear RNAs and EBV latent membrane antigen. The patient had no history of uterine leiomyoma or other neoplasms or clinical evidence of an immune defect. Leiomyoma should be included in the differential diagnosis of spindle cell tumours of the gallbladder and must be distinguished from leiomyosarcoma and the rare gastrointestinal stromal tumour-like neoplasms reported recently at this unusual anatomical site.


T cell receptor-proximal signals are sustained in peripheral microclusters and terminated in the central supramolecular activation cluster.
T cell receptor (TCR) signaling is initiated and sustained in microclusters; however, it's not known whether signaling also occurs in the TCR-rich central supramolecular activation cluster (cSMAC). We showed that the cSMAC formed by fusion of microclusters contained more CD45 than microclusters and is a site enriched in lysobisphosphatidic acid, a lipid involved in sorting ubiquitinated membrane proteins for degradation. Calcium signaling via TCR was blocked within 2 min by anti-MHCp treatment and 1 min by latrunculin-A treatment. TCR-MHCp interactions in the cSMAC survived these perturbations for 10 min and hence were not sufficient to sustain signaling. TCR microclusters were also resistant to disruption by anti-MHCp and latrunculin-A treatments. We propose that TCR signaling is sustained by stabilized microclusters and is terminated in the cSMAC, a structure from which TCR are sorted for degradation. Our studies reveal a role for F-actin in TCR signaling beyond microcluster formation.


Double-blind, placebo-controlled first in human study to investigate an oral vaccine aimed to elicit an immune reaction against the VEGF-Receptor 2 in patients with stage IV and locally advanced pancreatic cancer.
The investigational oral DNA vaccine VXM01 targets the vascular endothelial growth factor receptor 2 (VEGFR-2) and uses Salmonella typhi Ty21a as a vector. The immune reaction elicited by VXM01 is expected to disrupt the tumor neovasculature and, consequently, inhibit tumor growth. VXM01 potentially combines the advantages of anti-angiogenic therapy and active immunotherapy. This phase I trial examines the safety, tolerability, and immunological and clinical responses to VXM01. The randomized, placebo-controlled, double blind dose-escalation study includes up to 45 patients with locally advanced and stage IV pancreatic cancer. The patients will receive four doses of VXM01 or placebo in addition to gemcitabine as standard of care. Doses from 106 cfu up to 1010 cfu of VXM01 will be evaluated in the study. An independent data safety monitoring board (DSMB) will be involved in the dose-escalation decisions. In addition to safety as primary endpoint, the VXM01-specific immune reaction, as well as clinical response parameters will be evaluated. The results of this study shall provide the first data regarding the safety and immunogenicity of the oral anti-VEGFR-2 vaccine VXM01 in cancer patients. They will also define the recommended dose for phase II and provide the basis for further clinical evaluation, which may also include additional cancer indications. EudraCT No.: 2011-000222-29, NCT01486329, ISRCTN68809279.


Vascular reactivity in the spontaneously hypertensive stroke-prone rat. Effect of antihypertensive treatment.
This study investigated vascular responsiveness in stroke-prone spontaneously hypertensive rats (SHRSP) and the effect of antihypertensive treatment on this responsiveness. Weanling (4-week-old) male and female SHRSP and Wistar-Kyoto rats (WKY) received either the antihypertensive combination treatment of hydralazine plus hydrochlorothiazide in drinking water or tap water alone (controls) for 15 weeks. Whereas the antihypertensive combination prevented the development of hypertension in treated SHRSP (SHRSP-T), blood pressure remained unchanged in treated WKY (WKY-T). Femoral arterial smooth muscle responsiveness to KCl, norepinephrine, and calcium (in the presence of either 40 mM KCl or 1 microM norepinephrine) was not altered in SHRSP when compared with WKY. A significant increase in the sensitivity of femoral arteries to KCl and calcium (in the presence of 40 mM KCl) was seen, however, in SHRSP-T and WKY-T. An increased sensitivity to norepinephrine and calcium (in the presence of 1 microM norepinephrine) was seen only in SHRSP-T. Isoproterenol-induced relaxation was significantly attenuated in both SHRSP and SHRSP-T. Relaxation induced by sodium nitroprusside and calcium (membrane stabilization) was not different between the four groups. These results show that femoral arterial smooth muscle responsiveness to vasoconstrictor stimuli is not altered in SHRSP but that beta-adrenergic-mediated relaxation is attenuated. Antihypertensive treatment resulted in an enhanced responsiveness to these vasoconstrictor stimuli but had no effect on the relaxation properties of femoral arterial smooth muscle.


Inactivating Mutations in GT198 in Familial and Early-Onset Breast and Ovarian Cancers.
The human GT198 gene (gene symbol PSMC3IP) is located at chromosome 17q21, 470 kb proximal to BRCA1, a locus previously linked to breast and ovarian cancer predisposition. Its protein product (also known as TBPIP and Hop2) has been shown to regulate steroid hormone receptor-mediated gene activation and to stimulate homologous recombination in DNA repair. Here, we screened germline mutations in GT198 in familial and early-onset breast and ovarian cancer patients. We have identified 8 germline variants in a total of 212 index patients including reoccurring nonsense mutation c.310C>T (p.Q104X) and 5' UTR mutation c.-37A>T, each found in 2 unrelated families. Most identified index patients from cancer families had early onsets with a median age of 35 years. c.310C>T was absent in a total of 564 control individuals analyzed. GT198 gene amplification with an imbalanced mutant copy gain was identified in the blood DNA of one of the patients carrying c.310C>T. When tested, this truncating mutation abolished DNA damage-induced Rad51 foci formation. In addition, we have identified 15 somatic mutations in 2 tumors from 1 patient carrying germline mutation c.-37A>T. The presence of a somatic mutation on the wild-type allele showed that GT198 was biallelically mutated in the tumor. The somatic mutations identified near a splicing junction site caused defective alternative splicing and truncated the open reading frame. Therefore, distinct mutations may cause a similar consequence by truncating the full-length protein and inducing a loss of the wild type. Our study provides the first evidence of the presence of inactivating mutations in GT198 in familial and early-onset breast and ovarian cancer patients. Mutations in GT198, a gene regulating DNA repair, potentially contribute to an increased risk in familial breast and ovarian cancers.


Psychopathology, trauma and delinquency: subtypes of aggression and their relevance for understanding young offenders.
To examine the implications of an ontology of aggressive behavior which divides aggression into reactive, affective, defensive, impulsive (RADI) or "emotionally hot"; and planned, instrumental, predatory (PIP) or "emotionally cold." Recent epidemiological, criminological, clinical and neuroscience studies converge to support a connection between emotional and trauma related psychopathology and disturbances in the emotions, self-regulation and aggressive behavior which has important implications for diagnosis and treatment, especially for delinquent populations. Selective review of preclinical and clinical studies in normal, clinical and delinquent populations. In delinquent populations we observe an increase in psychopathology, and especially trauma related psychopathology which impacts emotions and self-regulation in a manner that hotly emotionally charged acts of aggression become more likely. The identification of these disturbances can be supported by findings in cognitive neuroscience. These hot aggressive acts can be delineated from planned or emotionally cold aggression. Our findings support a typology of diagnostic labels for disruptive behaviors, such as conduct disorder and oppositional defiant disorder, as it appears that these acts of hot emotional aggression are a legitimate target for psychopharmacological and other trauma specific interventions. The identification of this subtype of disruptive behavior disorders leads to more specific clinical interventions which in turn promise to improve hitherto unimpressive treatment outcomes of delinquents and patients with disruptive behavior.


The first ceratopsian dinosaur from South Korea.
In 2008, a new basal neoceratopsian was discovered in the Tando beds (Albian) of Tando Basin in South Korea. It represents the first ceratopsian dinosaur in the Korean peninsula and is assigned to Koreaceratops hwaseongensis gen. et sp. nov. Autapomorphies of Koreaceratops include very tall neural spines over five times higher than the associated centra in the distal caudals, and a unique astragalus divided into two fossae by a prominent craniocaudal ridge on the proximal surface. A phylogenetic analysis indicates that Koreaceratops is positioned between Archaeoceratops and all more derived neoceratopsians, and the elongation of caudal neural spines was an important derived character in non-ceratopsid neoceratopsians. The very tall caudal neural spines in Koreaceratops, Montanoceratops, Udanoceratops, Protoceratops, and Bagaceratops appear to be homoplasious, suggesting an independent adaptation, possibly for swimming. Skeletal evidence suggests that obligate quadrupedalism occurred gradually in neoceratopsians progressing from bipedal through facultative quadrupedalism, to complete quadrupedalism in Coronosauria.


Ni-Vanuatu health-seeking practices for general health and childhood diarrheal illness: results from a qualitative methods study.
A local perspective on diarrheal illness has been shown to enhance control strategies for diarrheal disease in traditional rural settings. We aimed to assess caregivers' understandings of childhood general and diarrheal illness, in one rural community in Vanuatu, to help formulate control strategies for preventing diarrheal disease. This was a descriptive study using qualitative analysis of responses to open-ended questions to provide a fuller understanding of illness. Thematic analysis with categories derived from medical anthropology was used to analyse responses and draw conclusions. Twenty-nine participants were interviewed; 22 were maternal responses, three were traditional practitioners, two were rural health care workers, one was a spiritual healer and one had a caregiver role. Respondents categorised illness as biomedical or traditional. Explanations of illness were enmeshed in and derived from both the traditional and biomedical system as the illness experience in the child under their care unfolded. Diarrheal severity influenced treatment selection and respondents expressed a preference for biomedical assistance. Respondents articulated a preference for biomedicine as the primary help-seeking resort for small children. Exclusive reliance on either traditional or biomedical options was uncommon. Local herbal remedies were the preferred home treatment when illness was known or mild, while oral rehydration therapy was used when accessing biomedical practitioners. Belief about diarrheal illness was influenced by traditional medicine and biomedicine. New evidence points to a growing preference for biomedicine as the first choice for severe childhood diarrheal illness. Diarrheal illness could be countered by maternal hand hygiene education at the medical dispensary and rural aid post.


Primary Care Practice Reports: Administrative Data Profiles for Identifying and Prioritizing Areas for Quality Improvement.
Information to help guide quality improvement activities in primary care should be readily available, routinely updated and include comparisons across groups, regions and jurisdictions. Primary care practice reports, developed jointly by the Institute for Clinical Evaluative Sciences and Health Quality Ontario, is one such effort. These data include practice demographics, the prevalence of common chronic conditions, the use of health services and measures of chronic disease prevention and management. All Ontario primary care physicians can register for the profiles online using a secure logon; the profiles are available only to them. Enhancements under development include new formats, targets and tools to support quality improvement.


A 3-week multimodal intervention involving high-intensity interval training in female cancer survivors: a randomized controlled trial.
To compare the effects of a 3-week multimodal rehabilitation involving supervised high-intensity interval training (HIIT) on female breast cancer survivors with respect to key variables of aerobic fitness, body composition, energy expenditure, cancer-related fatigue, and quality of life to those of a standard multimodal rehabilitation program. A randomized controlled trial design was administered. Twenty-eight women, who had been treated for cancer were randomly assigned to either a group performing exercise of low-to-moderate intensity (LMIE; n = 14) or a group performing high-intensity interval training (HIIT; n = 14) as part of a 3-week multimodal rehabilitation program. No adverse events related to the exercise were reported. Work economy improved following both HIIT and LMIE, with improved peak oxygen uptake following LMIE. HIIT reduced mean total body fat mass with no change in body mass, muscle or fat-free mass (best P < 0.06). LMIE increased muscle and total fat-free body mass. Total energy expenditure (P = 0.45) did not change between the groups, whereas both improved quality of life to a similar high extent and lessened cancer-related fatigue. This randomized controlled study demonstrates that HIIT can be performed by female cancer survivors without adverse health effects. Here, HIIT and LMIE both improved work economy, quality of life and cancer-related fatigue, body composition or energy expenditure. Since the outcomes were similar, but HIIT takes less time, this may be a time-efficient strategy for improving certain aspects of the health of female cancer survivors.


Hemosiderotic dermatofibroma: report of one case.
Dermatofibroma (DF) is a common benign fibrohistiocytic lesion which presents with a wide variety of clinicopathological features. Generally, the clinical diagnosis is easy, but differentiating it from other cutaneous tumors could be difficult in atypical cases and rare variants. We may find at least four different histopathological variants of DF; more than one of which may be present in a single tumor. Hemosiderotic DF is a variant composed of numerous small vessels, extravasated erythrocytes, and intra- and extracellular hemosiderotic deposits. The differential diagnosis may comprise melanoma as well as other melanocytic and nonmelanocytic tumors. We report the case of a 38-year-old man who presented with a hemosiderotic DF on the abdomen.


Genetic recombination in fused spheroplasts of Providence alcalifaciens.
Spheroplasts of Providence alcalifaciens strain P29 auxotrophs were prepared by combined treatment with glycine and lysozyme-EDTA. About 15% of spheroplasts had areas of cytoplasmic membrane exposed where cell wall was absent. The spheroplasts of different auxotrophs were mixed pairwise and fusion was attempted with polyethylene glycol or nascent calcium phosphate. After spheroplasts had regenerated to bacterial forms selection was made for recombinants. Recombinants arose at frequencies of 3.8 X 10(-6) to 1.7 X 10(-7) per spheroplast initially present, by both methods of fusion. The frequency was strongly dependent on the number of chromosomal loci used in selection. The possible order of five loci was determined and this corresponded to that on the closely related Proteus mirabilis chromosome. Control experiments excluded possibilities of auxotrophic reversion, conjugation, transformation, transfection or transduction as explanations of the results. Analysis of prototrophic clones yielded stable prototrophs or mixtures of stable prototrophs and stable recombinants. Parental types were not encountered. Unselected markers segregated among recombinants. It was concluded that the formation of recombinant bacteria was due to spheroplast fusion and that only stable products of the very temporary heteroploid state were haploid recombinants. The low frequency of recombination was ascribed to the limited number of spheroplasts with areas of exposed cytoplasmic membrane.


Supportive interventions for enhancing dietary intake in malnourished or nutritionally at-risk adults: a systematic review of nonrandomised studies.
Supportive interventions for enhancing dietary intake in malnourished or nutritionally at risk adults are frequently recommended. A recent systematic review of randomised controlled trials identified limited and poor quality evidence to support their use. Observational studies have been shown to compliment and extend their evidence. This review aimed to synthesise evidence from nonrandomised studies aiming to improve nutritional intake in nutritionally vulnerable individuals and to describe their effects on cost, nutritional, clinical and patient centred outcomes. Systematic searches of 10 electronic databases were undertaken to May 2013. Reference lists of identified studies and systematic reviews were scrutinised and hand searching of relevant meeting abstracts was undertaken. Titles and abstracts were reviewed, data extracted by two research-ers working independently and summarised using a structured narrative format. Forty-one studies (n = 3751 participants) were identified for inclusion. Interventions identified included changes to the organisation of nutritional care (n = 15), changes to the feeding environment (n = 11), modification to meals (n = 6), supplementation of meals (n = 7) and recipients of home delivered meals (n = 2). Eighteen of 23 studies reported improvements in nutritional intake; however, effects on nutritional status, clinical outcomes and costs were reported in few studies and findings were inconsistent. Eighteen studies reported patient experience, highlighting benefits to nutritional status most commonly as judged by the carers but the methods used lacked rigour. This systematic review describes a range of interventions that may be implemented in clinical practice. A limited range of outcomes are reported and it is difficult to draw any meaningful conclusions on the effect of the different methods.


The fractured landscape of RNA-seq alignment: the default in our STARs.
Many tools are available for RNA-seq alignment and expression quantification, with comparative value being hard to establish. Benchmarking assessments often highlight methods' good performance, but are focused on either model data or fail to explain variation in performance. This leaves us to ask, what is the most meaningful way to assess different alignment choices? And importantly, where is there room for progress? In this work, we explore the answers to these two questions by performing an exhaustive assessment of the STAR aligner. We assess STAR's performance across a range of alignment parameters using common metrics, and then on biologically focused tasks. We find technical metrics such as fraction mapping or expression profile correlation to be uninformative, capturing properties unlikely to have any role in biological discovery. Surprisingly, we find that changes in alignment parameters within a wide range have little impact on both technical and biological performance. Yet, when performance finally does break, it happens in difficult regions, such as X-Y paralogs and MHC genes. We believe improved reporting by developers will help establish where results are likely to be robust or fragile, providing a better baseline to establish where methodological progress can still occur.


The imitation game: Effects of social cues on 'imitation' are domain-general in nature.
Imitation has been hailed as 'social glue', facilitating rapport with others. Previous studies suggest that social cues modulate imitation but the mechanism of such modulation remains underspecified. Here we examine the locus, specificity, and neural basis of the social control of imitation. Social cues (group membership and eye gaze) were manipulated during an imitation task in which imitative and spatial compatibility could be measured independently. Participants were faster to perform compatible compared to incompatible movements in both spatial and imitative domains. However, only spatial compatibility was modulated by social cues: an interaction between group membership and eye gaze revealed more spatial compatibility for ingroup members with direct gaze and outgroup members with averted gaze. The fMRI data were consistent with this finding. Regions associated with the control of imitative responding (temporoparietal junction, inferior frontal gyrus) were more active during imitatively incompatible compared to imitatively compatible trials. However, this activity was not modulated by social cues. On the contrary, an interaction between group, gaze and spatial compatibility was found in the dorsolateral prefrontal cortex in a pattern consistent with reaction times. This region may be exerting control over the motor system to modulate response inhibition.


Silicon Reduces Black Sigatoka Development in Banana.
The effect of silicon (Si) uptake on the susceptibility of Musa acuminata to Mycosphaerella fijiensis was investigated in three experiments conducted under controlled conditions. Plants were grown in the presence of Si or not, in pots adapted for a hydroponic culture system or in pots filled with compost. The banana leaves were inoculated after 4 or 6 months of plant growth by spraying conidial suspensions or by brushing mycelia fragments. The disease progress over time was assessed using quantitative and qualitative scales. At the end of each experiment, disease severity was also analyzed using the image analysis software ASSESS. The Si concentration in the leaves of plants supplied with Si reached 10 to 28 g/kg of dry matter. The first symptoms appeared 18 days after inoculation. The disease developed more rapidly and more severely on banana plants grown without Si than on plants supplied with Si. The areas under the disease progress curve (AUDPCs) calculated for plants grown with Si were significantly lower than the AUDPCs for plants not supplied with Si, regardless of inoculation method. Thus, Si supply could be a valuable tool in integrated pest management against M. fijiensis by reducing the disease pressure on banana.


Fetal cocaine exposure causes persistent noradrenergic hyperactivity in rat brain regions: effects on neurotransmitter turnover and receptors.
Fetal cocaine exposure has been shown to cause neurobehavioral abnormalities related to ontogeny of catecholaminergic systems. In the current study, pregnant rats received cocaine (30 mg/kg s.c. daily) from gestational days 8 through 20 and markers of presynaptic and postsynaptic noradrenergic function were monitored in the offspring from birth to young adulthood. Animals in the cocaine group had normal body and brain region weights, and norepinephrine levels showed only minor differences from control values. However, fetal cocaine exposure produced marked noradrenergic hyperactivity, as assessed by norepinephrine turnover. The effect was selective for noradrenergic synapses, as dopamine levels and turnover were largely unaffected. The regional hierarchy of effects on norepinephrine turnover corresponded to the timetable of cell replication/differentiation, namely midbrain + brainstem > forebrain > cerebellum; the resemblance of this pattern to that for effects of glucocorticoids and hypoxia on noradrenergic cell differentiation suggests that cocaine alters development of noradrenergic activity by eliciting fetal hypoxia/ischemia and a consequent release of endogenous glucocorticoids. Effects of fetal cocaine exposure on development of adrenergic receptor subtypes (alpha-1, alpha-2 and beta receptors) were separable from those on noradrenergic presynaptic activity, exhibiting a different regional selectivity. Because cocaine exposure caused an increase in receptor binding superimposed on the increase in presynaptic activity, both factors are likely to contribute to a net synaptic hyperactivity and resultant behavioral teratogenesis.


TopicLens: Efficient Multi-Level Visual Topic Exploration of Large-Scale Document Collections.
Topic modeling, which reveals underlying topics of a document corpus, has been actively adopted in visual analytics for large-scale document collections. However, due to its significant processing time and non-interactive nature, topic modeling has so far not been tightly integrated into a visual analytics workflow. Instead, most such systems are limited to utilizing a fixed, initial set of topics. Motivated by this gap in the literature, we propose a novel interaction technique called TopicLens that allows a user to dynamically explore data through a lens interface where topic modeling and the corresponding 2D embedding are efficiently computed on the fly. To support this interaction in real time while maintaining view consistency, we propose a novel efficient topic modeling method and a semi-supervised 2D embedding algorithm. Our work is based on improving state-of-the-art methods such as nonnegative matrix factorization and t-distributed stochastic neighbor embedding. Furthermore, we have built a web-based visual analytics system integrated with TopicLens. We use this system to measure the performance and the visualization quality of our proposed methods. We provide several scenarios showcasing the capability of TopicLens using real-world datasets.


Calcineurin mutants render T lymphocytes resistant to cyclosporin A.
The immunosuppressants cyclosporin A (CsA) and FK506 have been widely used to prevent and treat graft rejection after human organ and tissue transplantations. CsA and FK506 associate with intracellular binding proteins (i.e., CsA with cyclophilin A and FK506 with FKBP12) to form protein/drug complexes that suppress the immune system by preventing activation of T cells in response to antigen presentation. The common target of CsA and FK506 is calcineurin, a Ca2+/calmodulin-regulated, serine/threonine-specific protein phosphatase that regulates the nuclear import of a transcription factor, NF-AT, required for expression of T cell activation genes. In previous studies, we identified calcineurin mutations that block binding by the cyclophilin A/CsA or FKBP12/FK506 complexes and thereby render yeast cells resistant to the antifungal effects of CsA or FK506. In this report, we demonstrate that the corresponding mutations in murine calcineurin render the T cell receptor signal transduction cascade CsA resistant in human Jurkat T cells. Our findings support the recently determined calcineurin X-ray crystal structure, provide evidence that calcineurin is the only CsA-sensitive component limiting signaling from the T cell receptor to the nucleus, and suggest a means to render cells and tissues resistant to the toxic side effects of CsA and FK506.


Adverse immunostimulation caused by impurities: The dark side of biopharmaceuticals.
Drug safety is an important issue, especially in the experimental phases of development. Adverse immunostimulation (AI) is sometimes encountered following treatment with biopharmaceuticals, which can be life-threatening if it results in a severe systemic inflammatory reaction. Biopharmaceuticals that unexpectedly induce an inflammatory response still enter the clinic, even while meeting all regulatory requirements. Impurities (of microbial origin) in biopharmaceuticals are an often-overlooked cause of AI. This demonstrates that the current guidelines for quality control and safety pharmacology testing are not flawless. Here, based on two case examples, several shortcomings of the guidelines are discussed. The most important of these are the lack of sensitivity for impurities, lack of testing for pyrogens other than endotoxin, and the use of insensitive animal species and biomarkers in preclinical investigations. Moreover, testing for the immunotoxicity of biopharmaceuticals is explicitly not recommended by the international guidelines. Publication of cases of AI is pivotal, both to increase awareness and to facilitate scientific discussions on how to prevent AI in the future.


Plasma trehalase activity and diabetes mellitus.
Trehalase is an enzyme which hydrolyzes the disaccharide trehalose, yielding glucose. It is widespread in nature and found in various human tissues as well as in human plasma. The synthesis and degradation of its substrate trehalose have been considered as being implicated in carbohydrate transport mechanisms. Trehalase activity has been examined in both normal subjects and diabetic patients. In the normal subjects, the frequency histogram of the enzyme activity is bimodal, indicating the existence of genetic polymorphism. The proposed model of a single autosomal locus with two alleles has been verified, with 27% of the population tested belonging to the "low-activity" phenotype and 73% being of the "high-activity" phenotype. Males have higher mean plasma trehalase activity than females. Apparently, the reverse appears to be the case in the diabetic subjects. The mean value for all nondiabetics and that of diabetics were computed and the difference was found to be statistically significant (F = 7.02, N1 = 3, N2 = 56, P less than 0.01). An experiment showed that neither the abnormally high concentration of glucose in diabetics nor any other constituent of the diabetic plasma caused an increase in plasma trehalase activity (t = 0.0724, P greater than 0.10). A Woolf and Haldane test to determine association of diabetes mellitus and plasma trehalase phenotype indicated a highly significant association with the high-activity phenotype (chi 2 = 18.5350, P less than 0.01). Thus the inference is that people with high plasma trehalase activity are more prone to develop diabetes mellitus than people with low enzyme activity.


Campylobacter pyloridis and associated gastritis: investigator blind, placebo controlled trial of bismuth salicylate and erythromycin ethylsuccinate.
An investigator blind trial was performed comparing bismuth salicylate, erythromycin ethylsuccinate, and placebo in the treatment of Campylobacter pyloridis associated gastritis in patients without peptic ulceration. Fifty patients fulfilled the study criteria. There was a strong correlation between the presence of C pyloridis and histologically confirmed gastritis. Clearance of organisms led to improvement of the gastritis. C pyloridis was cleared from 15 patients; of these, 13 had gastritis initially, which resolved in 12. Conversely, gastritis resolved in only four of 32 patients not cleared of organisms (p less than 0.0001). There was significantly greater improvement in endoscopic appearances in the patients cleared of C pyloridis compared with those whose infection persisted (p less than 0.001). In the three treatment groups organisms were cleared from 14 of 18 patients receiving the locally active bismuth salicylate, only one of 15 patients receiving erythromycin ethylsuccinate, and none of 17 patients taking placebo. These findings suggest that the ideal antimicrobial for the successful eradication of C pyloridis associated gastritis should be locally active, stable at low pH, and should penetrate gastric mucus. The resolution of gastritis and improvement in endoscopic appearances associated with clearance of C pyloridis support the view that these organisms may play a part in this condition.


[Cisapride does not modify prolonged Q-T interval in patients with liver cirrhosis].
Abnormal small bowel motility, observed in liver cirrhosis, can be reversed with cisapride. Since both cisapride and liver disease are associated with prolonged QT interval, the possibility of adverse cardiovascular effects might be expected with cisapride treatment in these patients. To evaluate QT interval and other electrocardiographic changes during long term treatment with cisapride in cirrhotic patients. Forty seven cirrhotic patients were studied. Electrocardiogram was recorded and the QT interval corrected according to Bazzett's formula was determined (normal value < 0.44 s). Seventeen patients were treated with cisapride, 10 mg tid for seven months and electrocardiographic controls were performed at the end of the treatment. The mean corrected QT interval was 0.46 +/- 0.03 s (range 0.4-0.53). 34 patients (64%) had QTc prolongation (0.47 +/- 0.02 s). Statistically significant higher values of QTc were observed in patients at Child Pugh stage B and C compared to stage A. No statistically significant difference according to the etiology of liver disease, were observed. No changes in mean QTc duration were observed during cisapride treatment. In spite that a prolonged QTc was a frequent finding in our series of selected patients, no cardiovascular adverse effects were observed with long term cisapride treatment.


Distinct roles for specific leptin receptor signals in the development of hypothalamic feeding circuits.
Circulating hormones influence multiple aspects of hypothalamic development and play a role in directing formation of neural circuits. Leptin is secreted by adipocytes and functions as a key developmental signal that promotes axon outgrowth from the arcuate nucleus (ARH) during a discrete developmental critical period. To determine the cellular mechanisms by which leptin impacts development of hypothalamic circuits, we examined roles for leptin receptor (LepRb) signals in neonatal mice. LepRb, ERK, and STAT3 signaling were required for leptin-stimulated neurite outgrowth from ARH explants in vitro. Neonatal mice with disrupted LepRb→ERK signaling displayed impaired ARH projections but were able to compensate by adulthood. LepRb→STAT3 signaling also plays a role in early circuit formation and controls the ultimate architecture of POMC, but not AgRP, projections. Thus, the developmental actions of leptin on feeding circuits are dependent on LepRb, and distinct signaling pathways are responsible for directing formation of NPY and POMC projections.


Channel selection for multispectral color imaging using binary differential evolution.
In multispectral color imaging, there is a demand to select a reduced number of optimal imaging channels to simultaneously speed up the image acquisition process and keep reflectance reconstruction accuracy. In this paper, the channel selection problem is cast as the binary optimization problem, and is consequently solved using a novel binary differential evolution (DE) algorithm. In the proposed algorithm, we define the mutation operation using a differential table of swapping pairs, and deduce the trial solutions using neighboring self-crossover. In this manner, the binary DE algorithm can well adapt to the channel selection problem. The proposed algorithm is evaluated on the multispectral color imaging system on both synthetic and real data sets. It is verified that high color accuracy is achievable by only using a reduced number of channels using the proposed method. In addition, as binary DE is a global optimization algorithm in nature, it performs better than the traditional sequential channel selection algorithm.


DNA barcoding in the Southeast Pacific marine realm: Low coverage and geographic representation despite high diversity.
The Southeast Pacific comprises two Large Marine Ecosystems, the Pacific Central-American Coastal and the Humboldt Current System; and is one of the less well known in the tropical subregions in terms of biodiversity. To address this, we compared DNA barcoding repositories with the marine biodiversity species for the Southeast Pacific. We obtained a checklist of marine species in the Southeast Pacific (i.e. Colombia, Ecuador, Chile, and Peru) from the Ocean Biodiversity Information System (OBIS) database and compared it with species available at the Barcoding of Life Data System (BOLD) repository. Of the 5504 species records retrieved from OBIS, 42% of them had at least one registered specimen in BOLD (including specimens around the world); however, only 4.5% of records corresponded to publicly available DNA barcodes including specimens collected from a Southeast Pacific country. The low representation of barcoded species does not vary much across the different taxonomic groups or within countries, but we observed an asymmetric distribution of DNA barcoding records for taxonomic groups along the coast, being more abundant for the Humboldt Current System than the Pacific Central-American Coastal. We observed high-level of barcode records with Barcode Index Number (BIN) incongruences, particularly for fishes (Actinopterygii = 30.27% and Elasmobranchii = 24.71%), reflecting taxonomic uncertainties for fishes, whereas for Invertebrates and Mammalia more than 85% of records were classified as data deficient or inadequate procedure for DNA barcoding. DNA barcoding is a powerful tool to study biodiversity, with a great potential to increase the knowledge of the Southeast Pacific marine biodiversity. Our results highlight the critical need for increasing taxonomic sampling effort, the number of trained taxonomic specialists, laboratory facilities, scientific collections, and genetic reference libraries.


The development and validation of a single SNaPshot multiplex for tiger species and subspecies identification--implications for forensic purposes.
The tiger (Panthera tigris) is currently listed on Appendix I of the Convention on the International Trade in Endangered Species of Wild Fauna and Flora; this affords it the highest level of international protection. To aid in the investigation of alleged illegal trade in tiger body parts and derivatives, molecular approaches have been developed to identify biological material as being of tiger in origin. Some countries also require knowledge of the exact tiger subspecies present in order to prosecute anyone alleged to be trading in tiger products. In this study we aimed to develop and validate a reliable single assay to identify tiger species and subspecies simultaneously; this test is based on identification of single nucleotide polymorphisms (SNPs) within the tiger mitochondrial genome. The mitochondrial DNA sequence from four of the five extant putative tiger subspecies that currently exist in the wild were obtained and combined with DNA sequence data from 492 tiger and 349 other mammalian species available on GenBank. From the sequence data a total of 11 SNP loci were identified as suitable for further analyses. Five SNPs were species-specific for tiger and six amplify one of the tiger subspecies-specific SNPs, three of which were specific to P. t. sumatrae and the other three were specific to P. t. tigris. The multiplex assay was able to reliably identify 15 voucher tiger samples. The sensitivity of the test was 15,000 mitochondrial DNA copies (approximately 0.26 pg), indicating that it will work on trace amounts of tissue, bone or hair samples. This simple test will add to the DNA-based methods currently being used to identify the presence of tiger within mixed samples.


Ribavirin for chronic hepatitis E virus infection in transplant recipients.
There is no established therapy for hepatitis E virus (HEV) infection. The aim of this retrospective, multicenter case series was to assess the effects of ribavirin as monotherapy for solid-organ transplant recipients with prolonged HEV viremia. We examined the records of 59 patients who had received a solid-organ transplant (37 kidney-transplant recipients, 10 liver-transplant recipients, 5 heart-transplant recipients, 5 kidney and pancreas-transplant recipients, and 2 lung-transplant recipients). Ribavirin therapy was initiated a median of 9 months (range, 1 to 82) after the diagnosis of HEV infection at a median dose of 600 mg per day (range, 29 to 1200), which was equivalent to 8.1 mg per kilogram of body weight per day (range, 0.6 to 16.3). Patients received ribavirin for a median of 3 months (range, 1 to 18); 66% of the patients received ribavirin for 3 months or less. All the patients had HEV viremia when ribavirin was initiated (all 54 in whom genotyping was performed had HEV genotype 3). At the end of therapy, HEV clearance was observed in 95% of the patients. A recurrence of HEV replication occurred in 10 patients after ribavirin was stopped. A sustained virologic response, defined as an undetectable serum HEV RNA level at least 6 months after cessation of ribavirin therapy, occurred in 46 of the 59 patients (78%). A sustained virologic response was also observed in 4 patients who had a recurrence and were re-treated for a longer period. A higher lymphocyte count when ribavirin therapy was initiated was associated with a greater likelihood of a sustained virologic response. Anemia was the main identified side effect and required a reduction in ribavirin dose in 29% of the patients, the use of erythropoietin in 54%, and blood transfusions in 12%. This retrospective, multicenter study showed that ribavirin as monotherapy may be effective in the treatment of chronic HEV infection; a 3-month course seemed to be an appropriate duration of therapy for most patients.


TNF-related apoptosis-inducing ligand (TRAIL) as a negative regulator of normal human erythropoiesis.
The impact of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on normal hematopoietic development was investigated using adult peripheral blood CD34(+) hematopoietic progenitor cells, induced to differentiate along the erythroid, megakaryocytic, granulocytic, and monocytic lineages by the addition of specific cytokine cocktails. TRAIL selectively reduced the number of erythroblasts, showing intermediate levels of glycophorin A (glycophorin A(interm)) surface expression, which appeared in liquid cultures supplemented with stem cell factor + interleukin 3 + erythropoietin at days 7-10. However, neither immature (day 4) glycophorin A(dim) erythroid cells nor mature (day 14) glycophorin A(bright) erythroblasts were sensitive to TRAIL-mediated apoptosis. Moreover, pre-exposure to TRAIL significantly decreased the number and size of erythroid colonies in semisolid assays. These adverse effects of TRAIL were selective for erythropoiesis, as TRAIL did not significantly influence the survival of cells differentiating along the megakaryocytic, granulocytic, or monocytic lineages. Furthermore, TRAIL was detected by Western blot analysis in lysates obtained from normal bone marrow mononuclear cells. These findings indicate that TRAIL acts in a lineage- and stage of differentiation-specific manner, as a negative regulator of normal erythropoiesis. (Blood. 2000;95:3716-3724)


Pain in multiple sclerosis: a systematic review of neuroimaging studies.
While pain in multiple sclerosis (MS) is common, in many cases the precise mechanisms are unclear. Neuroimaging studies could have a valuable role in investigating the aetiology of pain syndromes. The aim of this review was to synthesise and appraise the current literature on neuroimaging studies of pain syndromes in MS. We systematically searched PubMed and Scopus from their inception dates to the 2nd of April 2013. Studies were selected by predefined inclusion and exclusion criteria. Methodological quality was appraised. Descriptive statistical analysis was conducted. We identified 38 studies of variable methodology and quality. All studies but one used conventional structural magnetic resonance imaging, and the majority reported a positive association between location of demyelinating lesions and specific neuropathic pain syndromes. Most investigated headache and facial pain, with more common pain syndromes such as limb pain being relatively understudied. We identified a number of methodological concerns, which along with variable study design and reporting limit our ability to synthesise data. Higher quality studies were however less likely to report positive associations of lesion distribution to pain syndromes. Further high quality hypothesis-driven neuroimaging studies of pain syndromes in MS are required to clarify pain mechanisms, particularly for the commonest pain syndromes.