You really are too kind: implications regarding friendly submissiveness in trainee therapists.
To facilitate patient growth, therapists must immerse themselves in the patient's world while also being able to see what is needed for change. This process requires finding a delicate balance between supporting and pushing patients. Therapists in training are additionally tasked with incorporating supervisors' suggestions with their own views on what is needed to help their patients. Beginning therapists with tendencies to be overly accommodating may struggle to reconcile these competing demands. Thus, the aim of the present work is to explore how trainee friendly submissiveness (FS) interfaces with psychotherapy. Prior to training, clinical graduate trainee (n = 35) FS was assessed using the Inventory of Interpersonal Problems-32. Process and outcome data were then collected from each therapist's first training case. Specifically, each trainee was assigned an undergraduate student volunteer with whom they had four non-manualized therapy sessions over the academic semester. After the third session, patients and trainees completed questionnaires assessing session impact and the working alliance, and two expert raters coded third session videotapes for techniques. Following termination, patients rated the overall helpfulness of the therapy. Trainee FS was significantly negatively associated with patient-rated depth, alliance, and overall helpfulness with moderate effects. Findings from a mediation analysis further suggested that trainees with higher FS struggled to focus the therapy in a way that felt productive to patients. Implications for clinical training are discussed.


Anti-inflammatory and free radial scavenging activities of the constituents isolated from Machilus zuihoensis.
A new biflavonol glycoside, quercetin-3-O-β-D-glucopyranoside-(3'→O-3''')-quercetin-3-O-β-D-galactopyranoside (9), together with eight known compounds was isolated for the first time from the leaves of Machilus zuihoensis Hayata (Lauraceae). The structure of compound 9 was elucidated by various types of spectroscopic data analysis. Analysis of the biological activity assay found that compound 9 showed significant superoxide anion scavenging activity (IC₅₀ is 30.4 μM) and markedly suppressed LPS-induced high mobility group box 1 (HMGB-1) protein secretion in RAW264.7 cells. In addition, the HMGB-1 protein secretion was also inhibited by quercitrin (3), ethyl caffeate (6), and ethyl 3-O-caffeoylquinate (7) treatment. In the LPS-stimulated inducible nitric oxide synthase (iNOS) activation analysis, two known compounds, quercetin (1) and ethyl caffeate (6), were found to markedly suppress nitric oxide (NO) production (IC₅₀ value, 27.6 and 42.9 μM, respectively) in RAW264.7 cells. Additionally, it was determined that ethyl caffeate (6) down-regulated mRNA expressions of iNOS, IL-1β, and IL-10 in the LPS-treatment of RAW264.7 cells via a suppressed NF-kB pathway. These results suggested for the first time that the new compound 9 and other constituents isolated from M. zuihoensis have potential anti-inflammatory and superoxide anion scavenging effects. These constituents may be useful for treating various inflammatory diseases.


Genetic classification of populations using supervised learning.
There are many instances in genetics in which we wish to determine whether two candidate populations are distinguishable on the basis of their genetic structure. Examples include populations which are geographically separated, case-control studies and quality control (when participants in a study have been genotyped at different laboratories). This latter application is of particular importance in the era of large scale genome wide association studies, when collections of individuals genotyped at different locations are being merged to provide increased power. The traditional method for detecting structure within a population is some form of exploratory technique such as principal components analysis. Such methods, which do not utilise our prior knowledge of the membership of the candidate populations. are termed unsupervised. Supervised methods, on the other hand are able to utilise this prior knowledge when it is available.In this paper we demonstrate that in such cases modern supervised approaches are a more appropriate tool for detecting genetic differences between populations. We apply two such methods, (neural networks and support vector machines) to the classification of three populations (two from Scotland and one from Bulgaria). The sensitivity exhibited by both these methods is considerably higher than that attained by principal components analysis and in fact comfortably exceeds a recently conjectured theoretical limit on the sensitivity of unsupervised methods. In particular, our methods can distinguish between the two Scottish populations, where principal components analysis cannot. We suggest, on the basis of our results that a supervised learning approach should be the method of choice when classifying individuals into pre-defined populations, particularly in quality control for large scale genome wide association studies.


Association of perfluoroalkyl substances with gestational hypertension and preeclampsia in the MIREC study.
Perfluoroalkyl substances (PFAS) have been linked with a number of developmental, reproductive, hepatic, and cardiovascular health outcomes. However, the evidence for an association between PFAS and hypertensive disorders of pregnancy (including gestational hypertension and preeclampsia) is equivocal and warrants further investigation. To examine the relationship between background levels of perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), and perfluorohexane sulfonate (PFHxS) and the development of gestational hypertension or preeclampsia in a Canadian pregnancy cohort. We also explored the potential for effect modification according to fetal sex. Maternal plasma samples were collected in the first trimester from participants in the MIREC study and were analyzed for PFOA, PFOS, and PFHxS. Blood pressure was measured during each trimester. Gestational hypertension and preeclampsia were defined using the Society of Obstetricians and Gynaecologists of Canada guidelines. Logistic regression models were used to derive adjusted odds ratios (OR) and 95% confidence intervals (CI) for associations between PFAS concentrations (per doubling of concentration as well as according to tertiles) and gestational hypertension or preeclampsia. Linear mixed models were used to examine the association between PFAS concentrations and changes in blood pressure throughout pregnancy. Data from 1739 participants were analyzed. 90% of women were normotensive throughout pregnancy, 7% developed gestational hypertension without preeclampsia, and 3% developed preeclampsia. In the full analyses, neither PFOA nor PFOS were associated with gestational hypertension or preeclampsia. However, each doubling of PFHxS plasma concentration was associated with higher odds of developing preeclampsia (OR = 1.32; 95% CI: 1.03, 1.70). In addition, participants in the highest PFHxS tertile (1.4-40.0 μg/L) had higher odds of developing preeclampsia relative to those in the lowest tertile (OR = 3.06; 95% CI: 1.27, 7.39). In stratified analyses, this effect was only apparent among women carrying a female fetus (OR = 4.90; 95% CI: 1.02, 22.3). However, among women carrying a male fetus, both PFOS and PFHxS were associated with gestational hypertension, but not preeclampsia. Higher plasma concentrations of all three PFAS were associated with increases in diastolic blood pressure throughout pregnancy, and PFOA and PFHxS were also associated with systolic blood pressure. Discrepant findings were similarly revealed in analyses stratified by fetal sex. Higher levels of PFHxS were associated with the development of preeclampsia, but not gestational hypertension. Neither PFOA nor PFOS were associated with either outcome. However, we show, for the first time, that fetal sex may modify these associations, a finding which warrants replication and further study.


A rare cause of aeroallergen-induced anaphylaxis: horse allergy.
Animal epithelia and saliva contain a substantial amount of aeroallergen material, causing both acute onset of symptoms related to hypersensitivity, and chronic inflammation of the airways leading to allergic rhinitis or asthma. Although allergy to horses is generally considered to be more prevalent in rural areas, sensitization to horse allergens is more frequent than might be expected in urban residents without any direct or occupational exposure to horses. However, horse allergy is seldom mentioned among animal-induced allergic reactions. Anaphylaxis due to horse exposure is reported even more rarely. Here, a pediatric patient with a hypersensitivity reaction after horse allergen exposure is reported, and the definition of the type of reaction is discussed, as it may be the whole set of locally induced multisystemic reactions that resembles anaphylaxis.


Evaluation of the IT Infrastructure of the RESIST Study with the Evidence-Based CIPROS Checklist.
Efficient and powerful information systems are substantial to perform medical research projects successfully. Especially, translational medicine poses specific challenges to the corresponding IT infrastructure. The RESIST study is a translational research project in oncology where xenografts inform about patients second-line treatment. DBFORM, an in-house developed system, was used as EDC system. It was enhanced with project specific features. We demonstrate how the CIPROS checklist has the potential to optimize the related requirements engineering process. The CIPROS checklist consists of 72 items, organized within 12 Aspects/Topics and was developed to assess such patient registry software systems. In this paper we use the CIPROS checklist (1) to elucidate the projects requirements and (2) to assess systems features. The application of the CIPROS checklist to fix the RESIST project requirements and system features was successful. The interplay between (1) and (2) helped to accelerate the requirements engineering process and to set up a system suitable to perform the translational research project successful.


Evolution of DNA double-strand break repair by gene conversion: coevolution between a phage and a restriction-modification system.
The necessity to repair genome damage has been considered to be an immediate factor responsible for the origin of sex. Indeed, attack by a cellular restriction enzyme of invading DNA from several bacteriophages initiates recombinational repair by gene conversion if there is homologous DNA. In this work, we modeled the interaction between a bacteriophage and a bacterium carrying a restriction enzyme as antagonistic coevolution. We assume a locus on the bacteriophage genome has either a restriction-sensitive or a restriction-resistant allele, and another locus determines whether it is recombination/repair proficient or defective. A restriction break can be repaired by a co-infecting phage genome if one of them is recombination/repair proficient. We define the fitness of phage (resistant/sensitive and repair-positive/-negative) genotypes and bacterial (restriction-positive/-negative) genotypes by assuming random encounter of the genotypes, with given probabilities of single and double infections, and the costs of resistance, repair, and restriction. Our results show the evolution of the repair allele depends on b(1)/b(0), the ratio of the burst size b(1) under damage to host cell physiology induced by an unrepaired double-strand break to the default burst size b(0). It was not until this effect was taken into account that the evolutionary advantage of DNA repair became apparent.


Failure of translational repression in the phage f2 op3 mutant is not due to an altered coat protein-RNA interaction.
A secondary phenotype of the op3 mutant of RNA bacteriophage f2 is the absence of translational repression of the phage replicase gene by the phage coat protein. We have synthesized RNA fragments corresponding to the site of translational repression for both the wild type and the op3 mutant. Using a quantitative assay, we show that the affinity of the closely related R17 coat protein for the mutant and wild type RNA fragments is the same. In addition, we find that the op3 and R17 coat proteins bind to the wild type RNA fragment with essentially identical dissociation constants. Thus, the altered regulation of replicase protein synthesis in the op3 mutant does not appear to be due simply to a reduced affinity of the translational repressor for its target site.


Highly efficient multicolor multifocus microscopy by optimal design of diffraction binary gratings.
Multifocus microscopy (MFM) allows sensitive and fast three-dimensional imaging. It relies on the efficient design of diffraction phase gratings yielding homogeneous intensities in desired diffraction orders. Such performances are however guaranteed only for a specific wavelength. Here, we discuss a novel approach for designing binary phase gratings with dual color properties and improved diffraction efficiency for MFM. We simulate binary diffraction gratings with tunable phase shifts to explore its best diffraction performances. We report the design and fabrication of a binary array generator of 3 × 3 equal-intensity diffraction orders with 74% efficiency, 95% uniformity and dual color capability. The multicolor properties of this new design are highlighted by two-color MFM imaging. Finally, we discuss the basics of extending this approach to a variety of diffraction pattern designs.


Presenilins are essential for regulating neurotransmitter release.
Mutations in the presenilin genes are the main cause of familial Alzheimer's disease. Loss of presenilin activity and/or accumulation of amyloid-beta peptides have been proposed to mediate the pathogenesis of Alzheimer's disease by impairing synaptic function. However, the precise site and nature of the synaptic dysfunction remain unknown. Here we use a genetic approach to inactivate presenilins conditionally in either presynaptic (CA3) or postsynaptic (CA1) neurons of the hippocampal Schaeffer-collateral pathway. We show that long-term potentiation induced by theta-burst stimulation is decreased after presynaptic but not postsynaptic deletion of presenilins. Moreover, we found that presynaptic but not postsynaptic inactivation of presenilins alters short-term plasticity and synaptic facilitation. The probability of evoked glutamate release, measured with the open-channel NMDA (N-methyl-D-aspartate) receptor antagonist MK-801, is reduced by presynaptic inactivation of presenilins. Notably, depletion of endoplasmic reticulum Ca(2+) stores by thapsigargin, or blockade of Ca(2+) release from these stores by ryanodine receptor inhibitors, mimics and occludes the effects of presynaptic presenilin inactivation. Collectively, these results indicate a selective role for presenilins in the activity-dependent regulation of neurotransmitter release and long-term potentiation induction by modulation of intracellular Ca(2+) release in presynaptic terminals, and further suggest that presynaptic dysfunction might be an early pathogenic event leading to dementia and neurodegeneration in Alzheimer's disease.


Association of Environmental Arsenic Exposure, Genetic Polymorphisms of Susceptible Genes, and Skin Cancers in Taiwan.
Deficiency in the capability of xenobiotic detoxification and arsenic methylation may be correlated with individual susceptibility to arsenic-related skin cancers. We hypothesized that glutathione S-transferase (GST M1, T1, and P1), reactive oxygen species (ROS) related metabolic genes (NQO1, EPHX1, and HO-1), and DNA repair genes (XRCC1, XPD, hOGG1, and ATM) together may play a role in arsenic-induced skin carcinogenesis. We conducted a case-control study consisting of 70 pathologically confirmed skin cancer patients and 210 age and gender matched participants with genotyping of 12 selected polymorphisms. The skin cancer risks were estimated by odds ratio (OR) and 95% confidence interval (CI) using logistic regression. EPHX1 Tyr113His, XPD C156A, and GSTT1 null genotypes were associated with skin cancer risk (OR = 2.99, 95% CI = 1.01-8.83; OR = 2.04, 95% CI = 0.99-4.27; OR = 1.74, 95% CI = 1.00-3.02, resp.). However, none of these polymorphisms showed significant association after considering arsenic exposure status. Individuals carrying three risk polymorphisms of EPHX1 Tyr113His, XPD C156A, and GSTs presented a 400% increased skin cancer risk when compared to those with less than or equal to one polymorphism. In conclusion, GSTs, EPHX1, and XPD are potential genetic factors for arsenic-induced skin cancers. The roles of these genes for arsenic-induced skin carcinogenesis need to be further evaluated.


Carboxyl-terminal domain III of the delta' subunit of the DNA polymerase III holoenzyme binds delta.
The delta and delta' subunits are essential components of the DNA polymerase III holoenzyme, required for assembly and function of the DnaX-complex clamp loader (tau2gammadeltadelta'chipsi). The x-ray crystal structure of delta' contains three structural domains (Guenther, B., Onrust, R., Sali, A., O'Donnell, M., and Kuriyan, J. (1997) Cell 91, 335-345). In this study, we localize the delta-binding domain of delta' to a carboxyl-terminal domain III by quantifying the interaction of delta with a series of delta' fusion proteins lacking specific domains. Purification and immobilization of the fusion proteins were facilitated by the inclusion of a tag containing hexahistidine and a short biotinylation sequence. Both NH2- and COOH-terminal-tagged full-length delta' were soluble and had specific activities comparable with that of native delta'. delta and delta' form a 1:1 heterodimer with a dissociation constant (K(D)) of 5 x 10(-7) m determined by equilibrium sedimentation. The K(D) determined by surface plasmon resonance was comparable. Domain III alone bound delta at an affinity comparable to that of wild type delta', whereas proteins lacking domain III did not bind delta. Using a panel of domain-specific anti-delta' monoclonal antibodies, we found that two of the domain III-specific monoclonal antibodies interfered with delta-delta' interaction and abolished the replication activity of DNA polymerase-III holoenzyme.


Cell sensitivity to gravity.
Cultures of human lymphocytes exposed in microgravity to the mitogen concanavalin A showed less than 3 percent of the activation of ground controls. This result supports the hypothesis, based on simulations at low g and experiments at high g, that microgravity depresses whereas high gravity enhances cell proliferation rates. The effects of gravity are particularly strong in cells undergoing differentiation.


Digital pathology evaluation of complement C4d component deposition in the kidney allograft biopsies is a useful tool to improve reproducibility of the scoring.
Complement C4d component deposition in kidney allograft biopsies is an established marker of antibody-mediated rejection. In the Banff 07 classification of renal allograft pathology, semi-quantitative evaluation of the proportion of C4d-positive peritubular capilaries (PTC) is used. We aimed to explore the potential of digital pathology tools to obtain quantitative and reproducible measure of C4d deposition in the renal allograft tissue.34 routine kidney allograft biopsies immunohistochemically stained for C4d were included in the study and were evaluated by a qualified pathologist twice, recording an approximate percentage of positive PTC and glomerular area. The same slides were scanned by Aperio ScanScope scanner. Two layers of annotations were created: layer of glomeruli and the remaining non-glomerular area. Image analysis was performed with Aperio Positive Pixel Count algorithm to quantify the proportion of C4d-positive pixels in the area analysed. The percentage of positive (defined as 2+ and 3+) pixels in glomeruli and non-glomerular area was obtained and compared to the percentage of C4d-positive PTC and C4d-positive area of glomeruli recorded by the pathologist. The correlation of digital and manual C4d-positive area scoring in glomeruli was very high (r= 0.89, p<0.0001), while the correlation for non-glomerular (digital) and PTC (manual) area was moderate (r=0.60, p<0.001). The correlation between digital and manual evaluation of C4d in non-glomerular area after exclusion of C4d-positive arterioles from analysis did not improve substantially (r = 0.59, p < 0.001). Reproducibility of digital and manual results was evaluated. For C4d deposition in PTC, agreement between the first and the second digital C4d evaluation (after re-drawing annotations) was perfect (κ=0.96, CI 0.91÷1.00) while agreement between two subsequent manual C4d scorings was substantial (κ = 0.67, CI 0.47 ÷ 0.88). Similarly, for C4d deposition in the glomeruli, agreement of digital evaluation was perfect (κ=1) while for manual scorings it was substantial (κ = 0.76, CI 0.64 ÷ 0.88). Digital evaluation of C4d deposition in allograft kidney correlates with pathologist's scoring and exceeds the latter in reproducibilty. Therefore, it provides a useful tool to control for intraobserver and interobserver variability and may serve as quality assurance measure for allograft pathology diagnosis and research.


Influence of fecal sample storage on bacterial community diversity.
Previous studies have identified a correlation, either positive or negative, between specific stool bacteria strains and certain autoimmune diseases. These conflicting data may relate to sample collection. The aim of this work was to evaluate the influence of the collection parameters of time and temperature on bacterial community composition. Samples were taken from healthy children and immediately divided in 5 sub-samples. One sample was frozen immediately at -80 ° C, while the other aliquots were frozen 12, 24, 48, and 72h later DNA extracted from each sample was used to amplify the 16S rRNA with barcoded primers. The amplified products were pooled and partial 16S rRNA sequences were obtained by pyrosequencing. Person-to-person variability in community diversity was high. A list of those taxa that comprise at least 1% of the community was made for each individual. None of these were present in high numbers in all individuals. The Bacteroides were present in the highest abundance in three of four subjects. A total of 23,701 16S rRNA sequences were obtained with an average of 1,185 reads per sample with an average length of 200 bases. Although pyrosequencing of amplified 16S rRNA identified changes in community composition over time (~10%), little diversity change was observed at 12 hours (3.06%) with gradual changes occurring after 24 (8.61%), 48 (9.72%), and 72 h (10.14%), post collection.


Arginine metabolism regulates human erythroid differentiation through hypusination of eIF5A.
Metabolic programs contribute to hematopoietic stem and progenitor cell (HSPC) fate, but it is not known whether the metabolic regulation of protein synthesis controls HSPC differentiation. Here, we show that SLC7A1/cationic amino acid transporter 1-dependent arginine uptake and its catabolism to the polyamine spermidine control human erythroid specification of HSPCs via the activation of the eukaryotic translation initiation factor 5A (eIF5A). eIF5A activity is dependent on its hypusination, a posttranslational modification resulting from the conjugation of the aminobutyl moiety of spermidine to lysine. Notably, attenuation of hypusine synthesis in erythroid progenitors, by the inhibition of deoxyhypusine synthase, abrogates erythropoiesis but not myeloid cell differentiation. Proteomic profiling reveals mitochondrial translation to be a critical target of hypusinated eIF5A, and accordingly, progenitors with decreased hypusine activity exhibit diminished oxidative phosphorylation. This affected pathway is critical for eIF5A-regulated erythropoiesis, as interventions augmenting mitochondrial function partially rescue human erythropoiesis under conditions of attenuated hypusination. Levels of mitochondrial ribosomal proteins (RPs) were especially sensitive to the loss of hypusine, and we find that the ineffective erythropoiesis linked to haploinsufficiency of RPS14 in chromosome 5q deletions in myelodysplastic syndrome is associated with a diminished pool of hypusinated eIF5A. Moreover, patients with RPL11-haploinsufficient Diamond-Blackfan anemia as well as CD34+ progenitors with downregulated RPL11 exhibit a markedly decreased hypusination in erythroid progenitors, concomitant with a loss of mitochondrial metabolism. Thus, eIF5A-dependent protein synthesis regulates human erythropoiesis, and our data reveal a novel role for RPs in controlling eIF5A hypusination in HSPCs, synchronizing mitochondrial metabolism with erythroid differentiation.


Pro-NGF from Alzheimer's disease and normal human brain displays distinctive abilities to induce processing and nuclear translocation of intracellular domain of p75NTR and apoptosis.
The pro form of neurotrophic growth factor (pro-NGF), purified by chromatography from human Alzheimer's disease (AD)-affected brains (ADhbi-pro-NGF), has been shown to induce apoptotic cell death in neuronal cell cultures through its interaction with the p75 neurotrophin receptor (p75NTR). In the present work, we report that ADhbi-pro-NGF stimulates processing of p75NTR with alpha- and gamma-secretases, yielding a 20-kd intracellular domain (p75(ICD)) that translocates to the nucleus. This process was accompanied by delayed apoptosis. In AD, p75(ICD) was significantly increased in human entorhinal cortex. Although human frontal cortex has been described as showing a higher pro-NGF increase in AD, the increase in the entorhinal cortex paralleled p75NTR processing in its intracellular domain. In addition, pro-NGF isolated from AD-affected brains differed functionally from pro-NGF isolated from comparably aged control brains, with pro-NGF isolated from control brains being unstable and undergoing degradation to NGF when added to cell culture. As p75(ICD) and pro-NGF are both mediators of apoptosis and are both found in increased levels in the cerebral cortex in AD, the present data have implications for understanding neuronal degeneration in AD.


Platelet-Rich Plasma for Elbow Pathologies: a Descriptive Review of Current Literature.
Platelet-rich plasma is used in many orthopedic pathologies such as tendinopathies and ligament injuries. The clinical results reported in the scientific literature are, however, confusing. The aim of this manuscript is to provide a narrative literature review regarding the role of PRP in the most common elbow soft tissue pathologies. The response to PRP seems to be favorable when compared to steroid injection for pain management and for patient-reported outcomes in lateral epicondylitis. PRP injection does not seem to have the potential complications associated with a steroid injection such as skin atrophy, discoloration, and secondary tendon tears. Only a few manuscripts comparing the results of PRP treatment with either extracorporeal shockwave (ESW), dry needling, or even surgical treatments in lateral epicondylitis exist. The use of PRP in other elbow pathologies such as golfer's elbow, ulnar collateral ligament injury, and distal biceps and triceps pathology is examined in few studies, with unclear recommendations. Regarding elbow pathologies, PRP injections in tennis elbow seems to be the best-studied intervention. A major limitation in these studies is the significant heterogeneity in the methods used for preparing PRP, for example employing leukocyte-rich, leukocyte-poor preparations, PRP with or without activation, which makes the results of the studies difficult to compare. Results of this review show that more studies on larger cohorts, with comparable formulations, and with longer follow-up are required to give optimal suggestions concerning the use of PRP in elbow pathologies.


Cyclo-oligo-(1 → 6)-β-D-glucosamine based artificial channels for tunable transmembrane ion transport.
Unimolecular ion channels were designed by functionalization of a new type of cyclic oligosaccharides, cyclo-oligo-(1 → 6)-β-d-glucosamines, with pentabutylene glycol chains. Their ion transporting activity was tuned by varying oligomericity. A halide selectivity sequence, Cl(-) > Br(-) > I(-) was observed.


Disruption of IFT complex A causes cystic kidneys without mitotic spindle misorientation.
Intraflagellar transport (IFT) complexes A and B build and maintain primary cilia. In the mouse, kidney-specific or hypomorphic mutant alleles of IFT complex B genes cause polycystic kidneys, but the influence of IFT complex A proteins on renal development is not well understood. In the present study, we found that HoxB7-Cre-driven deletion of the complex A gene Ift140 from collecting ducts disrupted, but did not completely prevent, cilia assembly. Mutant kidneys developed collecting duct cysts by postnatal day 5, with rapid cystic expansion and renal dysfunction by day 15 and little remaining parenchymal tissue by day 20. In contrast to many models of polycystic kidney disease, precystic Ift140-deleted collecting ducts showed normal centrosomal positioning and no misorientation of the mitotic spindle axis, suggesting that disruption of oriented cell division is not a prerequisite to cyst formation in these kidneys. Precystic collecting ducts had an increased mitotic index, suggesting that cell proliferation may drive cyst expansion even with normal orientation of the mitotic spindle. In addition, we observed significant increases in expression of canonical Wnt pathway genes and mediators of Hedgehog and tissue fibrosis in highly cystic, but not precystic, kidneys. Taken together, these studies indicate that loss of Ift140 causes pronounced renal cystic disease and suggest that abnormalities in several different pathways may influence cyst progression.


Traumatic Childhood Experiences and Posttraumatic Stress Disorder Among Veterans in Substance Use Disorder Treatment.
Posttraumatic stress disorder (PTSD) dramatically increases the risk of both substance use disorder (SUD) and suicide in veterans. Military-related trauma, however, may not be the only or most significant trauma experienced by veterans. Trauma exposure is high among those joining the military. This study sought to identify the prevalence of five types of childhood trauma (emotional, physical, and sexual abuse and emotional and physical neglect) and three adult trauma symptom clusters (intrusive thoughts, avoidance, and hyperarousal) among veterans seeking SUD treatment and to clarify the associations between types of trauma and specific symptom clusters. Veterans at three Veterans Affairs (VA) SUD treatment facilities in the Midwest completed surveys at treatment entry (n1 = 195) and at 6-month follow-up (n2 = 138). Measures included the Childhood Trauma Questionnaire-Short Form and the PTSD Checklist, either a military or a civilian version, depending on whether the most traumatic event occurred in or out of the military. The prevalence of childhood trauma was high, ranging from 40.5% experiencing physical abuse down to 22.8% experiencing sexual abuse. At baseline, 60.2% of the military trauma group met criteria for PTSD, compared with 33.9% of the civilian trauma group, a significant difference, χ2(1, N = 195) = 14.46, p < .01. Childhood emotional and physical abuse were moderately associated with intrusion and hyperarousal in the military trauma group, but in the civilian trauma group a broader spectrum of childhood traumas were associated with a broader array of symptom clusters, including avoidance. At follow-up, symptoms improved and were less associated with childhood trauma. These findings illuminate the persistence of effects of childhood trauma and recommend more targeted PTSD treatments.


Predicting punching acceleration from selected strength and power variables in elite karate athletes: a multiple regression analysis.
This study investigated the relationship between punching acceleration and selected strength and power variables in 19 professional karate athletes from the Brazilian National Team (9 men and 10 women; age, 23 ± 3 years; height, 1.71 ± 0.09 m; and body mass [BM], 67.34 ± 13.44 kg). Punching acceleration was assessed under 4 different conditions in a randomized order: (a) fixed distance aiming to attain maximum speed (FS), (b) fixed distance aiming to attain maximum impact (FI), (c) self-selected distance aiming to attain maximum speed, and (d) self-selected distance aiming to attain maximum impact. The selected strength and power variables were as follows: maximal dynamic strength in bench press and squat-machine, squat and countermovement jump height, mean propulsive power in bench throw and jump squat, and mean propulsive velocity in jump squat with 40% of BM. Upper- and lower-body power and maximal dynamic strength variables were positively correlated to punch acceleration in all conditions. Multiple regression analysis also revealed predictive variables: relative mean propulsive power in squat jump (W·kg-1), and maximal dynamic strength 1 repetition maximum in both bench press and squat-machine exercises. An impact-oriented instruction and a self-selected distance to start the movement seem to be crucial to reach the highest acceleration during punching execution. This investigation, while demonstrating strong correlations between punching acceleration and strength-power variables, also provides important information for coaches, especially for designing better training strategies to improve punching speed.


Indications, Survivorship, and Clinical Outcomes of a Rotating Hinge Total Knee and Distal Femoral Arthroplasty System.
Rotating hinge knee prostheses (with or without distal femoral replacement) are indicated in cases of unreconstructible bony or soft tissue compromise. Despite their versatility, these implants have demonstrated high rates of mechanical failure. We aimed to review clinical outcomes of a novel hinged knee arthroplasty system. We reviewed all cases in a prospective database of hinged total knee arthroplasty (with and without distal femoral arthroplasty) involving a single manufacturer's implant system at a large tertiary-level academic health sciences center. We collected reasons for surgery, implant survivorship, reasons for revision, and clinical outcomes (Short Form 12, Western Ontario and McMaster Universities Osteoarthritis Index, and Knee Society Scores). Seventy-six cases of hinged knee implants using the knee system under investigation were performed (39 hinged total knee and 37 distal femoral replacements) between 2011 and 2018, inclusive. Indications for surgery varied, with second-stage reimplantation most common for rotating hinge and fracture most common for distal femoral arthroplasty. There were 6 revisions in the rotating hinge group (none for aseptic loosening) and 5 in the distal femoral arthroplasty group (1 for aseptic loosening). Average follow-up was 2.89 ± 2.09 years. Postoperative Short Form 12 (Physical Component), Western Ontario and McMaster Universities Osteoarthritis Index, and Knee Society Scores were 30.12 ± 10.17, 55.90 ± 21.51, and 115.62 ± 45.20, respectively. This novel hinged knee system is a highly durable option for complex and revision knee arthroplasty. Early failures tend to be secondary to infection. We observed only a single case of aseptic loosening. Although these early results are encouraging, ongoing follow-up is required to determine long-term prognosis in patients receiving this implant.


One-dimensional hard x-ray field retrieval using a moveable structure.
We present a technique that allows measuring the field of an x-ray line focus using far-field intensity measurements only. One-dimensional phase retrieval with transverse translation diversity is used to recover a hard x-ray beam focused by a compound kinoform lens. The reconstruction is found to be in good agreement with independent knife-edge scan measurements taken at separated planes. The approach avoids the need for measuring the beam profile at focus and allows narrower beams to be measured than the traditional knife-edge scan.


Non-thermal atmospheric plasma induces ROS-independent cell death in U373MG glioma cells and augments the cytotoxicity of temozolomide.
Non-thermal atmospheric plasma (NTAP) is an ionised gas produced under high voltage that can generate short-lived chemically active species and induce a cytotoxic insult in cancer cells. Cell-specific resistance to NTAP-mediated cytotoxicity has been reported in the literature. The aim of this study was to determine whether resistance against NTAP could be overcome using the human glioma cell line U373MG. Non-thermal atmospheric plasma was generated using a Dielectric Barrier Device (DBD) system with a maximum voltage output of 120 kV at 50 Hz. The viability of U373MG GBM cells and HeLa cervical carcinoma cells was determined using morphology, flow cytometry and cytotoxicity assays. Fluorescent probes and inhibitors were used to determine the mechanisms of cytotoxicity of cells exposed to the plasma field. Combinational therapy with temozolomide (TMZ) and multi-dose treatments were explored as mechanisms to overcome resistance to NTAP. Non-thermal atmospheric plasma decreased cell viability in a dose (time)-dependent manner. U373MG cells were shown to be resistant to NTAP treatment when compared with HeLa cells, and the levels of intracellular reactive oxygen species (ROS) quantified in U373MG cells were much lower than in HeLa cells following exposure to the plasma field. Reactive oxygen species inhibitor N-acetyl cysteine (NAC) only alleviated the cytotoxic effects in HeLa cells and not in the relatively NTAP-resistant cell line U373MG. Longer exposures to NTAP induced a cell death independent of ROS, JNK and caspases in U373MG. The relative resistance of U373MG cells to NTAP could be overcome when used in combination with low concentrations of the GBM chemotherapy TMZ or exposure to multiple doses. For the very first time, we report that NTAP induces an ROS-, JNK- and caspase-independent mechanism of cell death in the U373MG GBM cell line that can be greatly enhanced when used in combination with low doses of TMZ. Further refinement of the technology may facilitate localised activation of cytotoxicity against GBM when used in combination with new and existing chemotherapeutic regimens.


In vivo and in vitro studies on the toxicity of Hoechst 33342 (Ho342). Implications for employing Ho342 for the isolation of haematopoietic stem cells.
The fluorescence dye Hoechst 33342 (Ho342) is employed for isolating early haematopoietic cells and the aim of this study was to evaluate the in vivo and in vitro toxicity of this compound. First, by employing a murine model we studied the influence of this dye on the morphology of the different organs of animals that have been injected intravenously with increasing doses of Ho342. Accordingly, we found that Ho342 at relatively low doses (0,3 M) caused morphological changes in the spleen and lungs and at higher doses (1,5 & 6 M) damaged also the liver. In contrast, kidneys appear to be relatively resistant to this dye. Next, since Ho342 is employed for isolating early haematopoietic cells by FACS, we have been looking for potential toxicity of this dye against normal human haematopoietic progenitors. Accordingly, CD34+ cells isolated from cord blood (CB) samples were exposed to increasing doses of Ho342 (0-50 microM). We found that the low concentration of Ho342 (10 microM) recommended for isolating HSC significantly inhibited the clonogenecity of human erythroid progenitors (BFU-E). The higher doses of Ho342 have also been toxic against normal human myeloid progenitors (CFU-GM). This study shows that Ho342 could potentially damage human cells. This fact should be considered whenever Ho342 has to be employed for isolating living cells.


Early Identification of Reading Comprehension Difficulties.
Most research on early identification of reading disabilities has focused on word reading problems and little attention has been given to reading comprehension difficulties. In this study, we investigated whether measures of language ability and/or response to language intervention in kindergarten uniquely predicted reading comprehension difficulties in third grade. A total of 366 children were administered a battery of screening measures at the beginning of kindergarten and progress monitoring probes across the school year. A subset of children also received a 26-week Tier 2 language intervention. Participants' achievement in word reading was assessed at the end of second grade, and their performance in reading comprehension was measured as the end of third grade. Results showed that measures of language ability in kindergarten significantly added to the prediction of reading comprehension difficulties over and above kindergarten word reading predictors and direct measures of word reading in second grade. Response to language intervention also proved to be a unique predictor of reading comprehension outcomes. Findings are discussed in terms of their relevance for the early identification of reading disabilities.


Performance of lactating dairy cows fed whole cottonseed coated with gelatinized cornstarch.
The handling characteristics of whole cottonseed are improved by coating with gelatinized cornstarch, but limited information is available on the effects of feeding the coated cottonseed to lactating dairy cows. Thirty-six lactating Jersey cows were used in a crossover design trial with 4-wk experimental periods to evaluate the influence of coating whole cottonseed with 2.5% gelatinized cornstarch on dry matter intake, milk yield, and composition. Cows were fed diets containing 10.2% alfalfa-orchardgrass hay, 45.2% corn silage, 15.0% coated or uncoated whole cottonseed, and 29.6% concentrate for ad libitum consumption. Coating whole cottonseed with gelatinized cornstarch tended to reduce dry matter intake, which averaged 16.2 and 15.9 kg/d for uncoated and coated cottonseed, respectively. Milk yield and composition were similar for uncoated and coated cottonseed. The yield of energy-corrected milk per unit of dry matter consumed was greater with coated cottonseed. Cows fed coated cottonseed gained body weight, but cows fed uncoated cottonseed lost weight. Concentrations of plasma urea were similar among treatments; however, NEFA concentrations were lower for cows fed coated whole cottonseed. Results of this trial indicate that coating whole cottonseed with 2.5% gelatinized cornstarch does not alter its feeding value for lactating dairy cows.


Antibody performance in western blot applications is context-dependent.
An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13 000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data.


Indeterminate melanocytic proliferations of the conjunctiva.
The purpose of this study is to test the hypothesis that a subset of conjunctival melanocytic proliferations exists that cannot be reproducibly classified as benign, malignant, or indeterminate. Three groups of excisional biopsy specimens of conjunctival melanocytic proliferations were evaluated by 5 ophthalmic pathologists. These groups included lesions that were considered by the authors to represent benign (Group 1, n = 5), malignant (Group 2, n = 5) and indeterminate melanocytic proliferations (Group 3, n = 5). The panel classified the same sections in all 3 groups in a randomized, masked fashion, first without and then with a clinical history of patient age, sex and race. The kappa statistic (k) was used to quantify the degree of agreement among observers. There was strong concordance among the panel for both Group 1 (benign, k = 0.76) and Group 2 (malignant, k = 0.70) melanocytic proliferations. There was no concordance of the panel for Group 3 (indeterminate) lesions (k = -0.045). The concordance for Groups 1 and 2 and lack of concordance for Group 3 lesions were independent of knowledge of clinical history of age, sex, and race. A subset of melanocytic proliferations of the conjunctiva exists that cannot be reproducibly classified by pathologists as benign, malignant, or indeterminate.


Use of reconstructed computed tomography for the assessment of joint spaces in the canine elbow.
To assess the accuracy of reconstructed computed tomography for imaging canine elbow joint spaces. Computed tomography scans of eight cadaveric elbows were obtained and reconstructed computed tomography images were formatted in the dorsal and sagittal planes. Humeroradial and humeroulnar joint space measurements were obtained from these images. Intra-observer and inter-observer variations in joint space measurements were assessed, as was the effect of specimen positioning (inter-image variation). After freezing, four elbows were sectioned in the dorsal plane and four in the sagittal plane. In addition to visual comparison of the frozen sections with reconstructed computed tomography images, joint space measurements were obtained from frozen sections and compared with those from reconstructed computed tomography images. Variation was assessed using statistical calculations and graphical techniques. Both inter-image and intra-observer analyses revealed good agreement and low variation between data sets. Inter-observer correlation was only moderate, though variation was low. Visually, the reconstructed computed tomography images accurately reflected the frozen section anatomy. Agreement between frozen section and reconstructed computed tomography joint space measurements was good, with minimal variation. These results indicate that reconstructed computed tomography is capable of accurately imaging elbow joint spaces and precise joint space measurements can be obtained. Reconstructed computed tomography may be useful for determining joint space measurements and detecting elbow incongruencies in dogs with elbow dysplasia.


Knowing by heart: Visceral feedback shapes recognition memory judgments.
Although theories of emotion have long noted the importance of afferent feedback from the autonomic nervous system in generating feelings, there is a growing appreciation that this feedback may also play a role in shaping cognitive experiences. At present, little is known about its functional role in memory judgments. In the current study, we examined whether afferent cardiovascular feedback shapes recognition-memory decisions and experiences when previously encountered faces are being discriminated from novel ones. To investigate this possibility, we capitalized on the natural variation in baroreceptor mediated cardiovascular feedback that is associated with the cardiac cycle, synchronizing the brief presentation of memory probes during retrieval with individual heartbeats. In Experiments 1 and 2, we found that faces presented during cardiac systole (i.e., when visceral feedback is maximal) were more likely endorsed as "old" than those presented during cardiac diastole (i.e., when afferent feedback is minimal). This pattern was present for targets and lures, and held for faces with fearful or neutral expressions. Combining this manipulation with a remember/know procedure, Experiment 3 showed that the influence of afferent cardiovascular feedback is specific to trials on which participants report a feeling of familiarity without successful recollection of pertinent contextual detail. By revealing an influence of baroreceptor mediated cardiovascular feedback on familiarity, the current findings identify the functional role of a specific autonomic channel, previously implicated in emotion, in feeling states that pertain to memory experience.


Ultraviolet laser filaments for remote laser-induced breakdown spectroscopy (LIBS) analysis: applications in cultural heritage monitoring.
We report experiments with subpicosecond UV laser filaments for the remote analysis of samples related to objects of cultural heritage. The classic laser-induced breakdown spectroscopy (LIBS) technique finds new avenues through femtosecond filamentation, and dynamic remote LIBS becomes possible. Advantages such as self-regulated laser intensity deposition on the target suggest that there is significant potential for using UV femtosecond filaments for the remote analysis of sculpture and large monuments.


Biomechanical Evaluation of the Medial Stabilizers of the Patella.
Quantification of the biomechanical properties of each individual medial patellar ligament will facilitate an understanding of injury patterns and enhance anatomic reconstruction techniques by improving the selection of grafts possessing appropriate biomechanical properties for each ligament. To determine the ultimate failure load, stiffness, and mechanism of failure of the medial patellofemoral ligament (MPFL), medial patellotibial ligament (MPTL), and medial patellomeniscal ligament (MPML) to assist with selection of graft tissue for anatomic reconstructions. Descriptive laboratory study. Twenty-two nonpaired, fresh-frozen cadaveric knees were dissected free of all soft tissue structures except for the MPFL, MPTL, and MPML. Two specimens were ultimately excluded because their medial structure fibers were lacerated during dissection. The patella was obliquely cut to test the MPFL and the MPTL-MPML complex separately. To ensure that the common patellar insertion of the MPTL and MPML was not compromised during testing, only one each of the MPML and MPTL were tested per specimen (n = 10 each). Specimens were secured in a dynamic tensile testing machine, and the ultimate load, stiffness, and mechanism of failure of each ligament (MPFL = 20, MPML = 10, and MPTL = 10) were recorded. The mean ± SD ultimate load of the MPFL (178 ± 46 N) was not significantly greater than that of the MPTL (147 ± 80 N; P = .706) but was significantly greater than that of the MPML (105 ± 62 N; P = .001). The mean ultimate load of the MPTL was not significantly different from that of the MPML ( P = .210). Of the 20 MPFLs tested, 16 failed by midsubstance rupture and 4 by bony avulsion on the femur. Of the 10 MPTLs tested, 9 failed by midsubstance rupture and 1 by bony avulsion on the patella. Finally, of the 10 MPMLs tested, all 10 failed by midsubstance rupture. No significant difference was found in mean stiffness between the MPFL (23 ± 6 N/mm2) and the MPTL (31 ± 21 N/mm2; P = .169), but a significant difference was found between the MPFL and the MPML (14 ± 8 N/mm2; P = .003) and between the MPTL and MPML ( P = .028). The MPFL and MPTL had comparable ultimate loads and stiffness, while the MPML had lower failure loads and stiffness. Midsubstance failure was the most common type of failure; therefore, reconstruction grafts should meet or exceed the values reported herein. For an anatomic medial-sided knee reconstruction, the individual biomechanical contributions of the medial patellar ligamentous structures (MPFL, MPTL, and MPML) need to be characterized to facilitate an optimal reconstruction design.


Risk factors for malignancy in systemic sclerosis patients.
Systemic sclerosis (SSc) is an autoimmune connective tissue disease with multisystem involvement. An increased incidence of cancer in SSc patients compared with the general population has been reported in several reports. Our aims in this study were to determine the most common malignancies and to investigate the possible risk factors for the development of malignancy in patients with SSc. Three hundred forty SSc patients from 13 centers were included to the study. Data of the patients were obtained by evaluating their medical records retrospectively. A total of 340 patients with SSc were evaluated. Twenty-five of the patients had 19 different types of malignancy. Bladder cancer was the most common type of cancer with four patients and was followed by breast cancer with three patients, and cervix cancer and ovarian cancer with two patients each. Other types of cancers such as squamous cell skin cancer, adenocancer with an unknown origin, multiple myeloma, chronic myeloid leukemia, papillary thyroid cancer, larynx cancer, non-small cell lung cancer, follicular type non-Hodgkin lymphoma (NHL), endometrium cancer, colon cancer, uterus cancer, neuroendocrine tumor, glioblastoma multiforme, and soft tissue sarcoma were diagnosed in one patient each. The only cancer type that showed an association with cyclophosphamide dose was bladder carcinoma. Other malignancies did not show a correlation with age, sex, smoking, type and duration of the disease, autoantibodies, organ involvement, and dose and duration of cyclophosphamide therapy. Cancer may develop in any organ in patients with SSc. Continuous screening of the patients during a follow-up period is necessary for the early detection of the tumor development.


Invasive Validation of Antares, a New Algorithm to Calculate Central Blood Pressure from Oscillometric Upper Arm Pulse Waves.
Background: Antares is an algorithm for pulse wave analysis (PWA) by oscillometric blood pressure (BP) monitors in order to estimate central (aortic) blood pressure (cBP). Antares aims to enable brachial cuff-based BP monitors to be type II-devices, determining absolute cBP values independently of potential peripheral BP inaccuracies. The present study is an invasive validation of the Antares algorithm in the custo screen 400. Methods: We followed entirely the 2017 ARTERY protocol for validation of non-invasive cBP devices, the 2013 American National Standards Institute, Inc./Association for the Advancement of Medical Instrumentation/International Organization for Standardization (ANSI/AAMI/ISO) 81060-2 and 2018 AAMI/European Society of Hypertension (ESH)/ISO validation standard protocols. In total, 191 patients undergoing cardiac catheterization were included, of which 145 patients entered analysis. Invasive cBP recordings were compared to simultaneous non-invasive cBP estimations using the Antares algorithm, integrated into an oscillometric BP monitor. Results: Mean difference between invasive and non-invasively estimated systolic cBP was 0.71 mmHg with standard deviation of 5.95 mmHg, fulfilling highest validation criteria. Conclusion: Antares is the first algorithm for estimation of cBP that entirely fulfills the 2017 ARTERY and AAMI/ESH/ISO validation protocols. The Antares algorithm turns the custo screen 400 BP monitor into a type II-device. Integration of Antares into commercially available BP monitors could make it possible to measure PWA parameters in virtually every practice in future.


Can consumers cure healthcare?
The U.S. healthcare delivery system is in crises. Costs are too high and increasingly becoming unaffordable to federal and state governments, employers and consumers. Americans are dissatisfied with the current system and believe it should be fundamentally altered or rebuilt. A solution needs to be found, and it is not the single-payer system espoused by many in Washington and elsewhere. We believe consumers can cure healthcare if (a) professionals, providers and policy experts shift their mindset from treating diseases and conditions to taking a holistic approach to the caring of people, particularly Baby Boomers and their parents; (b) technology becomes widely available to increase engagement, personalize healthcare, share experiences, make better choices and embrace convenience and (c) a cost-effective and reimbursed primary care navigator (coordinator and/or health manager), consistent with the medical home concept espoused by the American Association of Family Practitioners (AAFP) becomes a central component of public policy.


Tau-Centric Multitarget Approach for Alzheimer's Disease: Development of First-in-Class Dual Glycogen Synthase Kinase 3β and Tau-Aggregation Inhibitors.
Several findings propose the altered tau protein network as an important target for Alzheimer's disease (AD). Particularly, two points of pharmacological intervention can be envisaged: inhibition of phosphorylating tau kinase GSK-3β and tau aggregation process. On the basis of this consideration and on our interest in multitarget paradigms in AD, we report on the discovery of 2,4-thiazolidinedione derivatives endowed with such a profile. 28 and 30 displayed micromolar IC50 values toward GSK-3β, together with the capacity of inhibiting AcPHF6 aggregation of 60% and 80% at 10 μM, respectively. In addition, they showed PAMPA-BBB permeability, together with a suitable cellular safety profile. 30 also displayed inhibition of both K18 and full-length tau aggregations. Finally, both compounds were able to improve cell viability in an okadaic acid-induced neurodegeneration cell model. To the best of our knowledge, 28 and 30 are the first balanced, nontoxic, dual-acting compounds hitting tau cascade at two different hubs.


Critical appraisal of MGMT in digestive NET treated with alkylating agents.
The efficacy of alkylating agents (temozolomide, dacarbazine, streptozotocin) in patients with advanced neuroendocrine tumors (NETs) has been well documented, especially in pancreatic NETs. Alkylating agents transfer methyl adducts on DNA bases. Among them, O6-methylguanine accounts for many of their cytotoxic effects and can be repaired by the O6-methylguanine-methyltransferase (MGMT). However, whether the tumor MGMT status could be a reliable biomarker of efficacy of alkylating agents in NETs is still a matter of debate. Herein, we sought to provide a critical appraisal of the role of the MGMT status in NETs. After reviewing the molecular mechanisms of repair of DNA damage induced by alkylating agents, we aimed to comprehensively review the methods of determination of the MGMT status and its impact on prognosis, prediction of objective response and progression-free survival in patients with advanced digestive NETs treated by alkylating agents. About half of pancreatic NETs are MGMT-deficient, as determined by impaired tumor MGMT expression or by MGMT promoter methylation. Overall, while published studies are heterogeneous and mostly limited in size, they advocate that MGMT deficiency may be a relevant biomarker for increased objective response rate, prolonged progression-fee survival and overall survival in patients with advanced NETs treated by alkylating agents. While these data require confirmation in prospective controlled studies, future research should focus on the standardization of MGMT status assessment. Additional mechanisms of repair of DNA damages induced by alkylating agents should be explored in order to identify biomarkers complementary to MGMT and targets for potential antitumor synergy, such as PARP.


Is our brain hardwired to produce God, or is our brain hardwired to perceive God? A systematic review on the role of the brain in mediating religious experience.
To figure out whether the main empirical question "Is our brain hardwired to believe in and produce God, or is our brain hardwired to perceive and experience God?" is answered, this paper presents systematic critical review of the positions, arguments and controversies of each side of the neuroscientific-theological debate and puts forward an integral view where the human is seen as a psycho-somatic entity consisting of the multiple levels and dimensions of human existence (physical, biological, psychological, and spiritual reality), allowing consciousness/mind/spirit and brain/body/matter to be seen as different sides of the same phenomenon, neither reducible to each other. The emergence of a form of causation distinctive from physics where mental/conscious agency (a) is neither identical with nor reducible to brain processes and (b) does exert "downward" causal influence on brain plasticity and the various levels of brain functioning is discussed. This manuscript also discusses the role of cognitive processes in religious experience and outlines what can neuroscience offer for study of religious experience and what is the significance of this study for neuroscience, clinicians, theology and philosophy. A methodological shift from "explanation" to "description" of religious experience is suggested. This paper contributes to the ongoing discussion between theologians, cognitive psychologists and neuroscientists.


Demonstration of an immune-mediated mechanism of penicillin-induced neutropenia and thrombocytopenia.
Severe neutropenia may be a more common complication of high-dose penicillin therapy than previously recognized. This report describes five such patients, one of whom also had thrombocytopenia. The neutrophil and platelet counts rapidly increased on stopping penicillin, and the bone-marrow, which was hypocellular in some cases, became normal. Further studies on one of these patients, using a fluorescent antiglobulin technique with paraformaldehyde-fixed cells, demonstrated a complement-fixing IgG penicillin antibody reacting with the patient's granulocytes and platelets in the presence of the drug. This suggested an immune mechanism similar to the well-recognized penicillin-induced immune haemolytic anaemia. The associated bone-marrow hypoplasia may also be due to antibody-mediated suppression of penicillin-coated precursor cells.


[Primary health care reform and implications for the organizational culture of Health Center Groups in Portugal].
The health sector's increasing complexity poses major challenges for administrators. There is considerable consensus on workforce quality as a key determinant of success for any health reform. This study aimed to explore the changes introduced by an action-training intervention in the organizational culture of the 73 executive directors of Health Center Groups (ACES) in Portugal during the primary health care reform. The study covers two periods, before and after the one-year ACES training, during which the data were collected and analyzed. The Competing Values Framework allowed observing that after the ACES action-training intervention, the perceptions of the executive directors regarding their organizational culture were more aligned with the practices and values defended by the primary health care reform. The study highlights the need to continue monitoring results over different time periods to elaborate further conclusions.


Reducing intrapartum-related neonatal deaths in low- and middle-income countries-what works?
Each year, 814,000 neonatal deaths and 1.02 million stillbirths result from intrapartum-related causes, such as intrauterine hypoxia. Almost all of these deaths are in low- and middle-income countries, where women frequently lack access to quality perinatal care and may delay care-seeking. Approximately 60 million annual births occur outside of health facilities, and most of these childbirths are without a skilled birth attendant. Conditions that increase the risk of intrauterine hypoxia--such as pre-eclampsia/eclampsia, obstructed labor, and low birth weight--are often more prevalent in low resource settings. Intrapartum-related neonatal deaths can be averted by a range of interventions that prevent intrapartum complications (eg, prevention and management of pre-eclampsia), detect and manage intrapartum problems (eg, monitoring progress of labor with access to emergency obstetrical care), and identify and assist the nonbreathing newborn (eg, stimulation and bag-mask ventilation). Simple, affordable, and effective approaches are available for low-resource settings, including community-based strategies to increase skilled birth attendance, partograph use by frontline health workers linked to emergency obstetrical care services, task shifting to increase access to Cesarean delivery, and simplified neonatal resuscitation training (Helping Babies Breathe(SM)). Coverage of effective interventions is low, however, and many opportunities are missed to provide quality care within existing health systems. In sub-Saharan Africa, recent health services assessments found only 15% of hospitals equipped to provide basic neonatal resuscitation. In the short term, intrapartum-related neonatal deaths can be substantially reduced by improving the quality of services for all childbirths that occur in health facilities, identifying and addressing the missed opportunities to provide effective interventions to those who seek facility-based care. For example, providing neonatal resuscitation for 90% of deliveries currently taking place in health facilities would save more than 93,000 newborn lives each year. Longer-term strategies must address the gaps in coverage of institutional delivery, skilled birth attendance, and quality by strengthening health systems, increasing demand for care, and improving community-based services. Both short- and long-term strategies to reduce intrapartum-related mortality should focus on reducing inequities in coverage and quality of obstetrical and perinatal care.


In vivo precision of conventional and digital methods of obtaining complete-arch dental impressions.
Digital impression systems have undergone significant development in recent years, but few studies have investigated the accuracy of the technique in vivo, particularly compared with conventional impression techniques. The purpose of this in vivo study was to investigate the precision of conventional and digital methods for complete-arch impressions. Complete-arch impressions were obtained using 5 conventional (polyether, POE; vinylsiloxanether, VSE; direct scannable vinylsiloxanether, VSES; digitized scannable vinylsiloxanether, VSES-D; and irreversible hydrocolloid, ALG) and 7 digital (CEREC Bluecam, CER; CEREC Omnicam, OC; Cadent iTero, ITE; Lava COS, LAV; Lava True Definition Scanner, T-Def; 3Shape Trios, TRI; and 3Shape Trios Color, TRC) techniques. Impressions were made 3 times each in 5 participants (N=15). The impressions were then compared within and between the test groups. The cast surfaces were measured point-to-point using the signed nearest neighbor method. Precision was calculated from the (90%-10%)/2 percentile value. The precision ranged from 12.3 μm (VSE) to 167.2 μm (ALG), with the highest precision in the VSE and VSES groups. The deviation pattern varied distinctly according to the impression method. Conventional impressions showed the highest accuracy across the complete dental arch in all groups, except for the ALG group. Conventional and digital impression methods differ significantly in the complete-arch accuracy. Digital impression systems had higher local deviations within the complete arch cast; however, they achieve equal and higher precision than some conventional impression materials.


Thermal infrared radiance simulation with aggregation modeling (TITAN): an infrared radiative transfer model for heterogeneous three-dimensional surface--application over urban areas.
The thermal infrared radiance simulation with aggregation modeling (TITAN) model, presented here, is an innovative transfer radiative code in the infrared domain (3-14 microm). It takes into account the three-dimensional (3D) structure of the landscape and simulates all the radiative components introduced by this 3D structure, which are due to the reflection and emission of walls and sloping roofs. Examples are given to illustrate the new opportunities offered by TITAN over urban areas. First, a phenomenological study is conducted at four wavelengths analyzing the relative effect of all the radiative contributors to the total signal. The same analysis is performed at bottom of atmosphere, which reveals an error occurring when a flat assumption is made (between 1% and 5%). In a second example, the directional effects at sensor level are simulated and show that the radiative temperature can vary by up to 10 K.


Methanol as a gasoline extender: a critique.
The tests conducted with the three vehicles at different emission control levels suggest that, in the area of fuel economy and emissions, potential benefits with methanol blends are related to carburetion and are only significant in the case of the rich-operating cars built before emission control standards were imposed. Theoretical considerations related to methanol's leaning effect on carburetion support this conclusion. Potential advantages for methanol in these areas are therefore continuously diminishing as the older cars leave the roads. At present, these older cars use only about one-fourth of the totalc motor gasoline consumed and, before methanol could be used on a large scale, this fraction would be much smaller. The use of methanol in gasoline would almost certainly create severe product quality problems. Water contamination could lead to phase separation in the distribution system and possibly in the car tank as well, and this would require additional investment in fuel handling and blending equipment. Excess fuel volatility in hot weather may also have adverse effects on car performance if the methanol blends include typical concentrations of butanes and pentanes. Removal of these light hydrocarbon components would detract from methanol's role as a gasoline extender and if current fuel volatility specifications were maintained, its use could lead to a net loss in the total available energy for use in motor fuels. Car performance problems associated with excessively lean operation would also be expected in the case of a significant proportion of late-model cars which are adjusted to operate on lean fuel-air mixtures. If methanol does become available in large quantities, these factors suggest that it would be more practical to use it for purposes other than those related to the extending of motor gasoline, such as for gas turbines used for electric power generation. In this case, the "pure" methanol would act as a cleanburning fuel, having none of the potentially severe product quality problems associated with its use in motor gasoline, while the fuel oil or natural gas cLirrently burned in these tuLrbines CotLild be diverted to other ulses.


Teaching trainers to incorporate evidence-based medicine (EBM) teaching in clinical practice: the EU-EBM project.
Evidence based medicine (EBM) is considered an integral part of medical training, but integration of teaching various EBM steps in everyday clinical practice is uncommon. Currently EBM is predominantly taught through theoretical courses, workshops and e-learning. However, clinical teachers lack confidence in teaching EBM in workplace and are often unsure of the existing opportunities for teaching EBM in the clinical setting. There is a need for continuing professional development (CPD) courses that train clinical trainers to teach EBM through on-the-job training by demonstration of applied EBM real time in clinical practice. We developed such a course to encourage clinically relevant teaching of EBM in post-graduate education in various clinical environments. We devised an e-learning course targeting trainers with EBM knowledge to impart educational methods needed to teach application of EBM teaching in commonly used clinical settings. The curriculum development group comprised experienced EBM teachers, clinical epidemiologists, clinicians and educationalists from institutions in seven European countries. The e-learning sessions were designed to allow participants (teachers) to undertake the course in the workplace during short breaks within clinical activities. An independent European steering committee provided input into the process. The curriculum defined specific learning objectives for teaching EBM by exploiting educational opportunities in six different clinical settings. The e-modules incorporated video clips that demonstrate practical and effective methods of EBM teaching in everyday clinical practice. The course encouraged focussed teaching activities embedded within a trainer's personal learning plan and documentation in a CPD portfolio for reflection. This curriculum will help senior clinicians to identify and make the best use of available opportunities in everyday practice in clinical situations to teach various steps of EBM and demonstrate their applicability to clinical practice. Once fully implemented, the ultimate outcome of this pilot project will be a European qualification in teaching EBM, which will be used by doctors, hospitals, professional bodies responsible for postgraduate qualifications and continuing medical education.


The big and small of it: how body size evolves.
Body size is a biological variable of fundamental importance and plays a central role in analyses of life history, sexual dimorphism, allometry, and natural and sexual selection. Yet, there remains a sizeable gulf in our understanding that lies between what we hypothesize influences change in size, from the point of view of ultimate causation, and what we know about how shifts in body size are regulated from a proximate perspective. I seek here to tie these two perspectives together, and specifically to argue that an understanding of the hormonal regulation of body size is necessary for constructing hypotheses regarding how body size evolves. Recent work using model organisms points to the insulin/insulin-like growth factor pathway as playing a key role in the regulation of growth, size, reproduction, and senescence. I review the role of various components of this pathway in regulating growth and size and illustrate the evidence for different ways in which these might work to generate differences in size in various organisms. Of particular interest are the tradeoffs between size and other life history traits produced by experimental alterations in this pathway. Recent work emphasizing the ways in which body size can be altered based on extrinsic factors provides the opportunity to link advances in uncovering the proximate bases of growth and size and offers an opportunity to frame new hypotheses regarding how variation in size evolves.


Racial disparities in automated speech recognition.
Automated speech recognition (ASR) systems, which use sophisticated machine-learning algorithms to convert spoken language to text, have become increasingly widespread, powering popular virtual assistants, facilitating automated closed captioning, and enabling digital dictation platforms for health care. Over the last several years, the quality of these systems has dramatically improved, due both to advances in deep learning and to the collection of large-scale datasets used to train the systems. There is concern, however, that these tools do not work equally well for all subgroups of the population. Here, we examine the ability of five state-of-the-art ASR systems-developed by Amazon, Apple, Google, IBM, and Microsoft-to transcribe structured interviews conducted with 42 white speakers and 73 black speakers. In total, this corpus spans five US cities and consists of 19.8 h of audio matched on the age and gender of the speaker. We found that all five ASR systems exhibited substantial racial disparities, with an average word error rate (WER) of 0.35 for black speakers compared with 0.19 for white speakers. We trace these disparities to the underlying acoustic models used by the ASR systems as the race gap was equally large on a subset of identical phrases spoken by black and white individuals in our corpus. We conclude by proposing strategies-such as using more diverse training datasets that include African American Vernacular English-to reduce these performance differences and ensure speech recognition technology is inclusive.


Ten-year outcome after rapid discontinuation of prednisone in adult primary kidney transplantation.
Rapid discontinuation of prednisone after kidney transplantation potentially allows for minimization of steroid-related side effects. Although intermediate-term data with rapid discontinuation of prednisone have been promising, concern still exists regarding long-term outcomes. The 10-year experience is reported herein. Between October 1, 1999 and December 31, 2010, 1241 adult primary kidney transplants (791 living donor and 450 deceased donor) were performed using a protocol in which prednisone is discontinued after postoperative day 5. The 10-year actuarial recipient and graft survival rates and prednisone-related side effects were studied. Ten-year actuarial patient survival was 71% for living donor transplants and 62% for deceased donor transplants; 10-year graft survival was 61% for living donor transplants and 51% for deceased donor transplants, and was comparable to 10-year Scientific Registry of Transplant Recipients national data. Ten-year death-censored graft survival was 79% for living donor transplants and 80% for deceased donor transplants. Ten-year acute rejection rates were 25% for deceased donor transplants and 31% for living donor transplants; 10-year chronic rejection (interstitial fibrosis/tubular atrophy) rates were 39% for deceased donor transplants and 47% for living donor transplants. For nondiabetic recipients of living donor or deceased donor allografts, the incidence of new-onset diabetes was significantly lower than in historical controls on prednisone (P<0.001). We also found significantly reduced rates of cataracts, avascular necrosis, and cytomegalovirus infection in some subgroups. Prednisone-related side effects can be minimized in a protocol incorporating rapid discontinuation of prednisone for maintenance immunosuppression. Ten-year patient and graft outcomes remain acceptable.


Transcranial magnetic stimulation dissociates prefrontal and parietal contributions to task preparation.
Cognitive control is thought to rely upon a set of distributed brain regions within frontoparietal cortex, but the functional contributions of these regions remain elusive. Here, we investigated the disruptive effects of transcranial magnetic stimulation (TMS) over the human prefrontal and parietal cortices in task preparation at different abstraction levels. While participants completed a task-switching paradigm that assessed the reconfiguration of task goals and response sets independently, TMS was applied over the left inferior frontal junction (IFJ) and over the left intraparietal sulcus (IPS) during task preparation. In Experiment 1, TMS over the IFJ caused interference with the updating of task goals, while leaving the updating of response sets unaffected. In Experiment 2, TMS over the IPS created the opposite pattern of results, perturbing only the ability to update response sets, but not task goals. Experiment 3 furthermore revealed that TMS over the IPS interfered with task goal updating when the pulses are delivered at a later point in time during preparation. This dissociation of abstract and action-related components not only reveals distinct cognitive control processes during task preparation, but also sheds new light on how prefrontal and parietal areas might work in concert to support flexible and goal-oriented control of behavior.


Developmental differences in posterior mesofrontal cortex recruitment by risky rewards.
Might increased risk taking in adolescence result in part from underdeveloped conflict-monitoring circuitry in the posterior mesofrontal cortex (PMC)? Adults and adolescents underwent functional magnetic resonance imaging during a monetary game of "chicken." As subjects watched ostensible winnings increase over time, they decided when to press a button to bank their winnings, knowing that if they did not stop pursuing money reward before a secret varying time limit, they would "bust" and either lose the money accrued on the current trial (low-penalty trials) or forfeit trial winnings plus a portion of previous winnings (high-penalty trials). Reward accrual at risk of low penalty (contrasted with guaranteed reward) activated the PMC in adults but not in adolescents. Across all subjects, this activation (1) correlated positively with age but negatively with risk exposure and (2) was greater when subjects busted on the previous low-penalty trial. Reward accrual at risk of high penalty was terminated sooner and recruited the PMC in both adults and adolescents when contrasted with guaranteed reward. Predecision PMC activation in the high-penalty trials was significantly reduced in trials when subjects busted. These data suggest that (1) under threat of an explicit severe penalty, recruitment of the PMC is similar in adolescents and adults and correlates with error avoidance, and (2) when potential penalties for a rewarding behavior are mild enough to encourage some risk taking, predecision PMC activation by a reward/risk conflict is sensitive to previous error outcomes, predictive of risk-aversive behavior in that trial, and underactive in adolescents.


Prognostic impact of pregnancy after breast cancer according to estrogen receptor status: a multicenter retrospective study.
We questioned the impact of pregnancy on disease-free survival (DFS) in women with history of breast cancer (BC) according to estrogen receptor (ER) status. A multicenter, retrospective cohort study in which patients who became pregnant any time after BC were matched (1:3) to patients with BC with similar ER, nodal status, adjuvant therapy, age, and year of diagnosis. To adjust for guaranteed time bias, each nonpregnant patient had to have a disease-free interval at least equal to the time elapsing between BC diagnosis and date of conception of the matched pregnant one. The primary objective was DFS in patients with ER-positive BC. DFS in the ER-negative cohort, whole population, and overall survival (OS) were secondary objectives. Subgroup analyses included DFS according to pregnancy outcome and BC-pregnancy interval. With a two-sided α = 5% and β = 20%, 645 ER-positive patients were required to detect a hazard ratio (HR) = 0.65. A total of 333 pregnant patients and 874 matched nonpregnant patients were analyzed, of whom 686 patients had an ER-positive disease. No difference in DFS was observed between pregnant and nonpregnant patients in the ER-positive (HR = 0.91; 95% CI, 0.67 to 1.24, P = .55) or the ER-negative (HR = 0.75; 95% CI, 0.51 to 1.08, P = .12) cohorts. However, the pregnant group had better OS (HR = 0.72; 95% CI, 0.54 to 0.97, P = .03), with no interaction according to ER status (P = .11). Pregnancy outcome and BC-pregnancy interval did not seem to impact the risk of relapse. Pregnancy after ER-positive BC does not seem to reduce the risk of BC recurrence.


Efficiency of high- and low-voltage pulse combinations for gene electrotransfer in muscle, liver, tumor, and skin.
Gene electrotransfer is gaining momentum as an efficient methodology for nonviral gene transfer. In skeletal muscle, data suggest that electric pulses play two roles: structurally permeabilizing the muscle fibers and electrophoretically supporting the migration of DNA toward or across the permeabilized membrane. To investigate this further, combinations of permeabilizing short high-voltage pulses (HV; hundreds of V/cm) and mainly electrophoretic long low-voltage pulses (LV; tens of V/cm) were investigated in muscle, liver, tumor, and skin in rodent models. The following observations were made: (1) Striking differences between the various tissues were found, likely related to cell size and tissue organization; (2) gene expression is increased, if there was a time interval between the HV pulse and the LV pulse; (3) the HV pulse was required for high electrotransfer to muscle, tumor, and skin, but not to liver; and (4) efficient gene electrotransfer was achieved with HV field strengths below the detectability thresholds for permeabilization; and (5) the lag time interval between the HV and LV pulses decreased sensitivity to the HV pulses, enabling a wider HV amplitude range. In conclusion, HV plus LV pulses represent an efficient and safe option for future clinical trials and we suggest recommendations for gene transfer to various types of tissues.


Atomic level description of the domain closure in a dimeric enzyme: thermus thermophilus 3-isopropylmalate dehydrogenase.
The domain closure associated with the catalytic cycle is described at an atomic level, based on pairwise comparison of the X-ray structures of homodimeric Thermus thermophilus isopropylmalate dehydrogenase (IPMDH), and on their detailed molecular graphical analysis. The structures of the apo-form without substrate and in complex with the divalent metal-ion to 1.8 Å resolution, in complexes with both Mn(2+) and 3-isopropylmalate (IPM), as well as with both Mn(2+) and NADH, were determined at resolutions ranging from 2.0 to 2.5 Å. Single crystal microspectrophotometric measurements demonstrated the presence of a functionally competent protein conformation in the crystal grown in the presence of Mn(2+) and IPM. Structural comparison of the various complexes clearly revealed the relative movement of the two domains within each subunit and allowed the identification of two hinges at the interdomain region: hinge 1 between αd and βF as well as hinge 2 between αh and βE. A detailed analysis of the atomic contacts of the conserved amino acid side-chains suggests a possible operational mechanism of these molecular hinges upon the action of the substrates. The interactions of the protein with Mn(2+) and IPM are mainly responsible for the domain closure: upon binding into the cleft of the interdomain region, the substrate IPM induces a relative movement of the secondary structural elements βE, βF, βG, αd and αh. A further special feature of the conformational change is the movement of the loop bearing the amino acid Tyr139 that precedes the interacting arm of the subunit. The tyrosyl ring rotates and moves by at least 5 Å upon IPM-binding. Thereby, new hydrophobic interactions are formed above the buried isopropyl-group of IPM. Domain closure is then completed only through subunit interactions: a loop of one subunit that is inserted into the interdomain cavity of the other subunit extends the area with the hydrophobic interactions, providing an example of the cooperativity between interdomain and intersubunit interactions.


GALT protein database: querying structural and functional features of GALT enzyme.
Knowledge of the impact of variations on protein structure can enhance the comprehension of the mechanisms of genetic diseases related to that protein. Here, we present a new version of GALT Protein Database, a Web-accessible data repository for the storage and interrogation of structural effects of variations of the enzyme galactose-1-phosphate uridylyltransferase (GALT), the impairment of which leads to classic Galactosemia, a rare genetic disease. This new version of this database now contains the models of 201 missense variants of GALT enzyme, including heterozygous variants, and it allows users not only to retrieve information about the missense variations affecting this protein, but also to investigate their impact on substrate binding, intersubunit interactions, stability, and other structural features. In addition, it allows the interactive visualization of the models of variants collected into the database. We have developed additional tools to improve the use of the database by nonspecialized users. This Web-accessible database (http://bioinformatica.isa.cnr.it/GALT/GALT2.0) represents a model of tools potentially suitable for application to other proteins that are involved in human pathologies and that are subjected to genetic variations.


Underlying Breast Cancer Risk and Menopausal Hormone Therapy.
The recent Collaborative Group on Hormonal Factors in Breast Cancer (CGHFBC) publication calculated the attributable risk of breast cancer from use of estrogen alone and estrogen plus a synthetic progestogen for less than 5 to 15 or more years of use. This CGHFB report calculated attributable risk based on their findings of relative risk from pooled data from 58 studies. Notably, neither the CGHFBC nor other previous studies have examined the effect of underlying risk of breast cancer on attributable risk. This omission prompted us to determine the magnitude of the effect of underlying risk on attributable risk in this perspective. Meaningful communication of the potential risk of menopausal hormonal therapy requires providing women with the estimated risk above their existing underlying risk (ie, attributable risk). Therefore, we have estimated attributable risks from the data published by the CGHFBC, taking into account varying degrees of underlying risk. Based on the Endocrine Society Guideline on Menopausal Hormone Therapy (MHT), we divided groups into 3 categories of risk: low (1.5%), intermediate (3.0%), and high (6.0%) underlying risk of breast cancer over 5 years. In women taking estrogen plus a synthetic progestogen for 5 to 9 years, the attributable risks of MHT increased from 12, to 42, to 85 additional women per 1000 in the low-, intermediate-, and high-risk groups, respectively. The attributable risks for estrogen alone were lower but also increased based on underlying risk. Notably, the attributable risks were amplified with duration of MHT use, which increased both relative risk and breast cancer incidence.


Evaluation of pump pulsation in respirable size-selective sampling: Part III. Investigation of European standard methods.
Lee et al. (Evaluation of pump pulsation in respirable size-selective sampling: part I. Pulsation measurements. Ann Occup Hyg 2014a;58:60-73) introduced an approach to measure pump pulsation (PP) using a real-world sampling train, while the European Standards (EN) (EN 1232-1997 and EN 12919-1999) suggest measuring PP using a resistor in place of the sampler. The goal of this study is to characterize PP according to both EN methods and to determine the relationship of PP between the published method (Lee et al., 2014a) and the EN methods. Additional test parameters were investigated to determine whether the test conditions suggested by the EN methods were appropriate for measuring pulsations. Experiments were conducted using a factorial combination of personal sampling pumps (six medium- and two high-volumetric flow rate pumps), back pressures (six medium- and seven high-flow rate pumps), resistors (two types), tubing lengths between a pump and resistor (60 and 90 cm), and different flow rates (2 and 2.5 l min(-1) for the medium- and 4.4, 10, and 11.2 l min(-1) for the high-flow rate pumps). The selection of sampling pumps and the ranges of back pressure were based on measurements obtained in the previous study (Lee et al., 2014a). Among six medium-flow rate pumps, only the Gilian5000 and the Apex IS conformed to the 10% criterion specified in EN 1232-1997. Although the AirChek XR5000 exceeded the 10% limit, the average PP (10.9%) was close to the criterion. One high-flow rate pump, the Legacy (PP=8.1%), conformed to the 10% criterion in EN 12919-1999, while the Elite12 did not (PP=18.3%). Conducting supplemental tests with additional test parameters beyond those used in the two subject EN standards did not strengthen the characterization of PPs. For the selected test conditions, a linear regression model [PPEN=0.014+0.375×PPNIOSH (adjusted R2=0.871)] was developed to determine the PP relationship between the published method (Lee et al., 2014a) and the EN methods. The 25% PP criterion recommended by Lee et al. (2014a), average value derived from repetitive measurements, corresponds to 11% PPEN. The 10% pass/fail criterion in the EN Standards is not based on extensive laboratory evaluation and would unreasonably exclude at least one pump (i.e. AirChek XR5000 in this study) and, therefore, the more accurate criterion of average 11% from repetitive measurements should be substituted. This study suggests that users can measure PP using either a real-world sampling train or a resistor setup and obtain equivalent findings by applying the model herein derived. The findings of this study will be delivered to the consensus committees to be considered when those standards, including the EN 1232-1997, EN 12919-1999, and ISO 13137-2013, are revised.


Parallel synthesis and screening of a solid phase carbohydrate library.
A solid phase carbohydrate library was synthesized and screened against Bauhinia purpurea lectin. The library, which contains approximately 1300 di- and trisaccharides, was synthesized with chemical encoding on TentaGel resin so that each bead contained a single carbohydrate. Two ligands that bind more tightly to the lectin than Gal-beta-1,3-GalNAc (the known ligand) have been identified. The strategy outlined can be used to identify carbohydrate-based ligands for any receptor; however, because the derivatized beads mimic the polyvalent presentation of cell surface carbohydrates, the screen may prove especially valuable for discovering new compounds that bind to proteins participating in cell adhesion.


Carnosine induces intestinal cells to secrete exosomes that activate neuronal cells.
Recently, we showed that imidazole dipeptide such as carnosine contained abundantly in chicken breast meat improves brain function in a double-blind randomized controlled trial. However, the underlying molecular mechanisms remain unknown. Here, we investigated whether carnosine activates intestinal epithelial cells and induces the secretion of factors that activate brain function. We focused on exosomes derived from intestinal epithelial cells as mediators of brain-gut interaction. Results showed that exosomes derived from Caco-2 cells treated with carnosine significantly induced neurite growth in SH-SY5Y cells. To clarify the molecular basis of this finding, we performed integrated analysis of microRNAs (miRNAs) with altered expression in exosomes in response to carnosine treatment and mRNAs with altered expression in target cells in response to exosome treatment to identify related miRNAs and their target genes. The combination of miR-6769-5p and its target gene ATXN1 was found to be involved in the exosome-induced activation of neuronal cells.


The evolving treatment paradigm in myelofibrosis.
Myelofibrosis (MF) is a BCR-ABL1-negative myeloproliferative neoplasm diagnosed de novo or developed from essential thrombocythemia (ET) or polycythemia vera (PV). Average survival of a patient with MF is 5-7 years. Disease complications include fatigue, early satiety, pruritus, painful splenic infarcts, infections and leukemic transformation. Allogeneic hematopoietic stem cell transplant (HSCT) is the only potentially curative option for MF, but carries a risk of treatment-related mortality and is reserved for the few high-risk patients fit enough to endure the procedure. Other traditional therapies are palliative and supported by few randomized, controlled trials; thus, novel treatment strategies are needed. Discovery of the Janus kinase 2 (JAK2) gain-of-function mutation, JAK2V617F, in the majority (50-60%) of patients with MF led to increased understanding of the biology underlying MF and the development of JAK2 inhibitors to treat MF. Recent Food and Drug Administration (FDA) approval of the first JAK2 inhibitor, ruxolitinib, signaled a new era for treatment of MF. Additional JAK2 inhibitors, such as SAR302503, may become commercially available in the near future, and their distinct pharmacologic and efficacy profiles will help determine their use across the patient population. Data on JAK2 inhibitors, their role in an evolving treatment paradigm, and future directions for treatment of MF are discussed.


Tiotropium suppresses acetylcholine-induced release of chemotactic mediators in vitro.
The driving force in the progression of COPD is the development of exacerbations which are mostly the result of excessive inflammation. Bronchodilatators play an important role in the treatment of COPD. The reported reduction in exacerbation rates in COPD is due to the inhibition of vagal-mediated bronchoconstriction and mucus secretion. However, recent studies have highlighted the existence of muscarinic receptors on inflammatory cells and we have explored the possibility that tiotropium bromide might also inhibit neutrophil migration. We analysed the influence of tiotropium on the release of neutrophil chemotactic activity in response to acetylcholine (ACh) and the expression of muscarinic receptors on human alveolar macrophages (AM), A549 cells, MonoMac6 cells, and human lung fibroblasts. We found significant levels of all muscarinic receptor subtypes on all analysed cells except the fibroblasts. Fibroblasts expressed predominantly M2, receptors and did not release chemotactic activity. AM, A549 cells, and MonoMac6 cells released chemotactic active mediators after incubation with ACh. The secretion could be suppressed by more than 70% after coincubation with tiotropium. Tiotropium alone did not influence the granulocyte migration. Most of the chemotactic activity could be attributed to leukotriene B4 (LTB4). The release of interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) was not induced by ACh. From this, we suggest that the suppression of the Ach-mediated release of chemotactic substances like LTB4 modulates the inflammatory reaction. This may contribute to the decreased rate of exacerbations in COPD, which was observed in clinical trials.