Deep Learning-Based Algorithm for Detecting Aortic Stenosis Using Electrocardiography.
Background Severe, symptomatic aortic stenosis (AS) is associated with poor prognoses. However, early detection of AS is difficult because of the long asymptomatic period experienced by many patients, during which screening tools are ineffective. The aim of this study was to develop and validate a deep learning-based algorithm, combining a multilayer perceptron and convolutional neural network, for detecting significant AS using ECGs. Methods and Results This retrospective cohort study included adult patients who had undergone both ECG and echocardiography. A deep learning-based algorithm was developed using 39 371 ECGs. Internal validation of the algorithm was performed with 6453 ECGs from one hospital, and external validation was performed with 10 865 ECGs from another hospital. The end point was significant AS (beyond moderate). We used demographic information, features, and 500-Hz, 12-lead ECG raw data as predictive variables. In addition, we identified which region had the most significant effect on the decision-making of the algorithm using a sensitivity map. During internal and external validation, the areas under the receiver operating characteristic curve of the deep learning-based algorithm using 12-lead ECG for detecting significant AS were 0.884 (95% CI, 0.880-0.887) and 0.861 (95% CI, 0.858-0.863), respectively; those using a single-lead ECG signal were 0.845 (95% CI, 0.841-0.848) and 0.821 (95% CI, 0.816-0.825), respectively. The sensitivity map showed the algorithm focused on the T wave of the precordial lead to determine the presence of significant AS. Conclusions The deep learning-based algorithm demonstrated high accuracy for significant AS detection using both 12-lead and single-lead ECGs.


Phospholipase activity on N-acyl phosphatidylethanolamines is critically dependent on the N-acyl chain length.
We have recently shown that an endogenous phospholipase A2 from bovine erythrocytes does not hydrolyse NAPEs (N-acyl L-alpha-phosphatidylethanolamines), which accumulate remarkably in this system [Florin-Christensen, Suarez, Florin-Christensen, Wainszelbaum, Brown, McElwain and Palmer (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 7736-7741]. Here we investigate the causes underlying this resistance. N-acylation of PE (L-alpha-phosphatidylethanolamine) results in alteration of charge, head-group volume and conformation, the last two features depending on the N-acyl chain length. To evaluate each effect separately, we synthesized NAPEs with selected N-acyl chain length. We found that phospholipase A2 has considerable activity against N-acetyl PE, but is poorly active against N-butanoyl PE and only marginally active against N-hexanoyl PE, whereas the activity is completely lost when N-hexadecanoyl PE is presented as a substrate. On the other hand, N-hexanoyl PE does not inhibit phospholipase A2 activity, suggesting that this substrate fails to enter the hydrophobic channel. Phospholipase C presents a similar, but less sharp pattern. Molecular dynamics simulations of the polar head group of selected NAPEs reveal a substantially increased conformational variability as the N-acyl chain grows. This larger conformational space represents an increased impairment limiting the access of these molecules to the active site. Our data indicate that, whereas a change in charge contributes to diminished activity, the most relevant effects come from steric hindrance related to the growth of the N-acyl chain.


Notch signaling controls the differentiation of transporting epithelia and multiciliated cells in the zebrafish pronephros.
Epithelial tubules consist of multiple cell types that are specialized for specific aspects of organ function. In the zebrafish pronephros, multiciliated cells (MCCs) are specialized for fluid propulsion, whereas transporting epithelial cells recover filtered-blood solutes. These cell types are distributed in a ;salt-and-pepper' fashion in the pronephros, suggesting that a lateral inhibition mechanism may play a role in their differentiation. We find that the Notch ligand Jagged 2 is expressed in MCCs and that notch3 is expressed in pronephric epithelial cells. Morpholino knockdown of either jagged 2 or notch3, or mutation in mind bomb (in which Notch signaling is impaired), dramatically expands ciliogenic gene expression, whereas ion transporter expression is lost, indicating that pronephric cells are transfated to MCCs. Conversely, ectopic expression of the Notch1a intracellular domain represses MCC differentiation. Gamma-secretase inhibition using DAPT demonstrated a requirement for Notch signaling early in pronephric development, before the pattern of MCC differentiation is apparent. Strikingly, we find that jagged 2 knockdown generates extra cilia and is sufficient to rescue the kidney cilia mutant double bubble. Our results indicate that Jagged 2/Notch signaling modulates the number of multiciliated versus transporting epithelial cells in the pronephros by way of a genetic pathway involving repression of rfx2, a key transcriptional regulator of the ciliogenesis program.


The developmental origins of osteoporotic fracture.
Undernutrition and other adverse influences arising in fetal life or immediately after birth have a permanent effect on body structure, physiology and metabolism. Evidence is now accumulating from human studies that programming of bone growth might be an important contributor to the later risk of osteoporotic fracture. Body weight in infancy is a determinant of adult bone mineral content, as well as of the basal levels of activity of the GH/IGF-1 and HPA axes, and recent work has suggested a central role for vitamin D. Epidemiological studies have suggested that maternal smoking and nutrition during pregnancy influence intrauterine skeletal mineralization. Finally, childhood growth rates have been directly linked to the risk of hip fracture many decades later. Further work is needed to use this approach to develop novel therapeutic and preventative strategies to reduce the burden of osteoporotic fractures in the population.


Sustainable Development for Whom and How? Exploring the Gaps between Popular Discourses and Ground Reality Using the Mexican Jatropha Biodiesel Case.
In the last decade, jatropha-based bioenergy projects have gotten significant attention as a solution to various social, economic, and environmental problems. Jatropha's popularity stemmed out from different discourses, some real and some perceived, in scientific and non-scientific literature. These discourses positioned jatropha as a crop helpful in producing biodiesel and protecting sustainability by reducing greenhouse gas emissions compared to fossil fuels and increasing local, rural development by creating jobs. Consequently, many countries established national policies that incentivized the establishment of jatropha as a bioenergy feedstock crop. In this paper, we explore the case of jatropha bioenergy development in Yucatan, Mexico and argue that the popular discourse around jatropha as a sustainability and rural development tool is flawed. Analyzing our results from 70 semi-structured interviews with community members belonging to a region where plantation-scale jatropha projects were introduced, we found that these projects did not have many significant social sustainability benefits. We conclude from our case that by just adding bioenergy projects cannot help achieve social sustainability in rural areas alone. In ensuring social sustainability of bioenergy projects, future policymaking processes should have a more comprehensive understanding of the rural socioeconomic problems where such projects are promoted and use bioenergy projects as one of the many solutions to local problems rather than creating such policies based just on popular discourses.


Evaluation of ricin A chain-containing immunotoxins directed against CD19 and CD22 antigens on normal and malignant human B-cells as potential reagents for in vivo therapy.
Ricin A chain-containing immunotoxins (IT-As) specific for the human B-cell antigens, CD22 and CD19, were constructed using the monoclonal antibodies, HD6 and HD37, respectively. IT-As were prepared by coupling intact antibodies, F(ab')2, or Fab' fragments to native or chemically deglycosylated ricin A chain. The IT-As were then evaluated for cytotoxicity to normal and neoplastic human B-cells in vitro with the major objective of appraising their suitability for in vivo therapy of human B-cell tumors. The IT-As prepared with both the HD6 and HD37 antibodies were specifically toxic to normal B-cells and to most of the neoplastic B-cell lines tested. However, the IT-As prepared from HD6 were generally more potent than those prepared from HD37. On Daudi cells, to which the two antibodies bound in similar numbers and with similar affinities, IT-As prepared with intact HD6 antibody or its Fab' fragment were 10-fold and 1.5- to 4-fold more potent, respectively, than the corresponding HD37 IT-As. The IT-As constructed from intact HD6 antibody and native or deglycosylated A chain reduced protein synthesis in Daudi cells by 50% at a concentration of 1.2 X 10(-11) M indicating that they were only 5-fold less toxic to the cells than ricin itself. Intact HD37 IT-As produced equivalent inhibition of protein synthesis at 1.5 X 10(-10) M. With both antibodies, IT-As constructed from the Fab' fragments were 10- to 20-fold less potent than their intact antibody counterparts. Different neoplastic B-cell lines varied in sensitivity to the IT-As. In most cases, their sensitivity correlated with the levels of CD19 and CD22 antigens expressed. Neither HD6 nor HD37 IT-As affected the ability of normal human bone marrow cells to form granulocyte-macrophage colony-forming units in soft agar, suggesting that both antigens are absent from these progenitor cells. Examination of sections of frozen human tissues using immunoperoxidase staining procedures indicated that the antibodies did not bind to a panel of normal tissues lacking B-lymphocytes. These results suggest that HD6 and HD37 IT-As are candidates for in vivo therapy in humans with certain B-cell tumors. However, HD6 IT-As are more potent, reduce protein synthesis more completely, and hence appear to be the ITs of choice for treating tumors expressing the CD22 antigen.


The neuronal replicator hypothesis.
We propose that replication (with mutation) of patterns of neuronal activity can occur within the brain using known neurophysiological processes. Thereby evolutionary algorithms implemented by neuro- nal circuits can play a role in cognition. Replication of structured neuronal representations is assumed in several cognitive architectures. Replicators overcome some limitations of selectionist models of neuronal search. Hebbian learning is combined with replication to structure exploration on the basis of associations learned in the past. Neuromodulatory gating of sets of bistable neurons allows patterns of activation to be copied with mutation. If the probability of copying a set is related to the utility of that set, then an evolutionary algorithm can be implemented at rapid timescales in the brain. Populations of neuronal replicators can undertake a more rapid and stable search than can be achieved by serial modification of a single solution. Hebbian learning added to neuronal replication allows a powerful structuring of variability capable of learning the location of a global optimum from multiple previously visited local optima. Replication of solutions can solve the problem of catastrophic forgetting in the stability-plasticity dilemma. In short, neuronal replication is essential to explain several features of flexible cognition. Predictions are made for the experimental validation of the neuronal replicator hypothesis.


Localization of cellular src mRNA during development and in the differentiated bipolar neurons of the adult neural retina in Xiphophorus.
The expression of the c-src gene in embryonic and adult tissue of the teleost fish Xiphophorus helleri was analyzed by in-situ hybridization. The highly conserved fish c-src gene was found to be expressed at high levels in midterm embryos, where c-src mRNA was localized in developing neurons of the sensory layer of the differentiating retina and in the developing brain. In adult tissues the expression of c-src was found to persist in certain cell types of the brain and the neural retina, especially in the bipolar cells of the inner nuclear layer, which are postmitotic, fully differentiated mature neurons. Thus c-src in Xiphophorus appears to be a developmentally regulated proto-oncogene which is important for neuronal differentiation during organogenesis, but whose persistence of expression in certain terminally differentiated neurons strongly suggests a particular maintenance function for c-src in these cells as well.


Oxidative DNA damage and defence gene expression in the mouse lung after short-term exposure to diesel exhaust particles by inhalation.
Exposure to diesel exhaust particles (DEP) is suspected to contribute to lung cancer and cardiopulmonary diseases. In recent years generation of reactive oxygen species capable of inducing cellular oxidative stress has been in focus as one of the underlying mechanisms behind the genotoxic effects of particles. However, the role of the antioxidative defence system still needs to be clarified, especially in relation to low-dose DEP exposures. The aim of this study was to characterize the effects of short-term exposure to DEP in terms of DNA damage and expression of key response genes towards oxidative stress in lungs of mice. Mice were exposed by inhalation to 20 or 80 mg/m3 DEP inhaled as either a single dose, or four lower doses (5 and 20 mg/m3) inhaled on four consecutive days. Our results indicate that HO-1 mRNA expression in lung tissue was up-regulated after both types of DEP exposures, whereas OGG1 expression was only up-regulated after repeated exposures. The level of oxidative DNA damage in terms of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) was increased in the lung tissue after a single exposure, whereas increased levels of DNA strand breaks was observed in bronchoalveolar lavage cells after repeated DEP exposures. The levels of 8-oxodG and OGG1 mRNA in lung tissue were mirror images. This suggests that after repeated exposures, up-regulation of DNA repair counteracts an increased rate of 8-oxodG formation leaving the steady state level of 8-oxodG in DNA unchanged. In conclusion, this study indicates that a single high dose of DEP generates 8-oxodG in lung tissue, whereas the same dose inhaled as four low-exposures may up-regulate the antioxidative defence system and protect against generation of 8-oxodG.


Vaginal Ring Use in a Phase 3 Microbicide Trial: A Comparison of Objective Measures and Self-reports of Non-adherence in ASPIRE.
This analysis compares self-reports of product use with objective measures of non-adherence-quarterly plasma dapivirine levels and monthly residual dapivirine (DPV) levels in used rings-in MTN-020/ASPIRE, a phase 3 trial of a monthly DPV vaginal ring among women aged 18-45 years in Malawi, South Africa, Uganda and Zimbabwe. For participants on active product (N = 1211) we assessed self-reported monthly non-adherence, as measured by (1) whether the ring was ever out, and out for ≥ 12 h in the previous month and, (2) by a self-rating scale assessing ability to keep the vaginal ring inserted, and compared the self-reports to two biomarkers of non-use separately and as a composite measure. For this analysis, a plasma DPV value ≤ 95 pg/ml and residual ring ≥ 23.5 mg were used to classify non-adherence (i.e. the ring never being in the vagina the previous month). Compared to self-reports, non-adherence was found to be substantially higher for the composite measure as well as its two components, an indication that ring removal was likely underreported in the trial. The discrepancy between the self-report measure of ring outage and the composite indicator was greater for those aged 18-21 than for those older, evidence that younger women are more likely to underreport non-adherence. Despite underreporting of non-adherence, self-reports of the ring never being out were significant in predicting the composite objective measure. Furthermore, the association between the self-rating scale and the objective measure was in the expected direction and significant, although 11% of those 18-21 and 7% of those 22+ who rated their ability to keep the ring inserted as good, very good or excellent in the 4 weeks prior to exit were considered non-adherent according to the objective measure. This analysis indicates that while self-reports are significantly associated with objective measures of adherence in the ASPIRE trial, they were inflated-more so by those younger-and therefore may have limited utility identifying those who have challenges using products as directed. ClinicalTrials.gov number NCT01617096.


Drosophila melanogaster genes for U1 snRNA variants and their expression during development.
We have cloned and characterized a complete set of seven U1-related sequences from Drosophila melanogaster. These sequences are located at the three cytogenetic loci 21D, 82E, and 95C. Three of these sequences have been previously studied: one U1 gene at 21D which encodes the prototype U1 sequence (U1a), one U1 gene at 82E which encodes a U1 variant with a single nucleotide substitution (U1b), and a pseudogene at 82E. The four previously uncharacterized genes are another U1b gene at 82E, two additional U1a genes at 95C, and a U1 gene at 95C which encodes a new variant (U1c) with a distinct single nucleotide change relative to U1a. Three blocks of 5' flanking sequence similarity are common to all six full length genes. Using specific primer extension assays, we have observed that the U1b RNA is expressed in Drosophila Kc cells and is associated with snRNP proteins, suggesting that the U1b-containing snRNP particles are able to participate in the process of pre-mRNA splicing. We have also examined the expression throughout Drosophila development of the two U1 variants relative to the prototype sequence. The U1c variant is undetectable by our methods, while the U1b variant exhibits a primarily embryonic pattern reminiscent of the expression of certain U1 variants in sea urchin, Xenopus, and mouse.


Effect of prior intensive therapy in type 1 diabetes on 10-year progression of retinopathy in the DCCT/EDIC: comparison of adults and adolescents.
The aim of this study was to examine differences between adolescents and adults in persistence of the benefits of intensive therapy 10 years after completion of the Diabetes Control and Complications Trial (DCCT). During the Epidemiology of Diabetes Interventions and Complications (EDIC) study, progression of retinopathy from DCCT closeout to EDIC year 10 was evaluated in 1,055 adults and 156 adolescents. During 10 years of follow-up, HbA(1c) (A1C) was similar between original intensive (INT) and conventional (CON) groups and between former adolescents and adults. At EDIC year 10, adults in the former INT group continued to show slower progression of diabetic retinopathy than those in the CON group (adjusted hazard reduction 56%, P < 0.0001), whereas in adolescents this beneficial effect had disappeared (32%, P = 0.13). Seventy-nine percent of observed differences in the prolonged treatment effect between adults and adolescents at year 10 were explained by differences in mean A1C during DCCT between adolescents and adults (8.9 vs. 8.1%), particularly between INT adolescents and adults (8.1 vs. 7.2%). Prior glycemic control during DCCT is vital for the persistence of the beneficial effects of INT therapy 10 years later. Lowering A1C to as close to normal as safely possible without severe hypoglycemia and starting as early as possible should be attempted for all subjects with type 1 diabetes. These results underscore the importance of maintaining A1C at target values for as long as possible because the benefits of former INT treatment wane over time if A1C levels rise.


Cost-effectiveness of Endovascular Therapy for Acute Ischemic Stroke: A Systematic Review of the Impact of Patient Age.
Purpose To determine the impact of patient age on the cost-effectiveness of endovascular therapy (EVT) in addition to standard care (SC) in large-vessel-occlusion stroke for patients aged 50 to 100 years in the United States. Materials and Methods A decision-analytic Markov model was used to estimate direct and indirect lifetime costs and quality-adjusted life years (QALYs). Age-dependent input parameters were obtained from the literature. Deterministic and probabilistic sensitivity analysis for age at index stroke were used. The willingness-to-pay (WTP) was set to thresholds of $50 000, $100 000, and $150 000 per QALY. The study applied a U.S. setting for health care and societal perspectives. Incremental costs and effectiveness were derived from deterministic and probabilistic sensitivity analysis. Acceptability rates at different WTP thresholds were determined. Results EVT+SC was the dominant strategy in patients aged 50 to 79 years. The highest incremental effectiveness (2.61 QALYs) and cost-savings (health care perspective, $99 555; societal perspective, $146 385) were obtained in 50-year-old patients. In octogenarians (80-89 years), EVT+SC led to incremental QALYs at incremental costs with acceptability rates of more than 85%, more than 99%, and more than 99% at a WTP of $50 000, $100 000, and $150 000 per QALY, respectively. In nonagenarians (90-99 years), acceptability rates at a WTP of $50 000 per QALY dropped but stayed higher than 85% and higher than 95% at thresholds of $100 000 and $150 000 per QALY. Conclusion Using contemporary willingness-to-pay thresholds in the United States, endovascular therapy in addition to standard care reduces lifetime costs for patients up to 79 years of age and is cost-effective for patients aged 80 to 100 years.


Matrix multiplication by optical methods: experimental verification.
Matrix multiplication has been performed by optical analog methods using the scheme of Heinz et al. The most critical element was F(a), the spatial filter corresponding to the two-dimensional Fourier transform of a. In the operation c = b x a, errors in the c elements were 1-2% when the a elements were all approximately the same. Very large errors occurred when the a elements differed significantly, and these were used directly [instead of the F(a) derived values] in the numerical comparisons. These errors were attributed to the limited linear range of the amplitude transmittance characteristic of the film used. Suggestions for future work are made.


A novel metal-free amidosulfenylation of alkenes leading to β-azolyl sulfides.
A novel and environmentally benign method for C-S and C-N bond formation by the direct amidosulfenylation of alkenes has been developed under metal-free conditions. Various alkenes and azoles were transformed into the corresponding β-azolyl sulfides in ionic liquids. The wide substrate scope, good functional group tolerance, and ease of operation make this reaction attractive for the synthesis of nitrogen- and sulfur-containing molecules.


Religion and physical health among older Israeli Jews: findings from the SHARE-Israel study.
Despite decades of research on religious determinants of health, this subject has not been systematically investigated within Jewish populations, in Israel or the diaspora. The present paper is part of a series of studies using large-scale population data sources to map the impact of religiousness on the physical and mental health of Jews. To identify religious predictors of physical health in a national probability sample of older Israeli Jews. The data derive from the Israeli sample of the Survey of Health, Ageing and Retirement in Europe (SHARE), a cross-national survey program involving nearly a dozen nations. The Israeli sample comprises 1287 Jewish respondents aged 50 or over. Outcome measures include single-item assessments of self-rated health, long-term health problems, and activity limitation, as well as validated measures of diagnosed chronic diseases, physical symptoms, and activities of daily living (ADL) and instrumental ADL (IADL). Recent synagogue attendance is a significant predictor of better health for six of the seven health measures, even after adjusting for age and several other covariates and mediators, including measures of health-related behavior and social support. Prayer, by contrast, is inversely associated with health according to five measures, perhaps reflecting its use as a coping mechanism for individuals with health problems. This study presents modest evidence of a salutary effect of Jewish religiousness on this population of older adults. Religiousness, in the form of synagogue participation, was seen to serve a protective function, and prayer a coping function.


Comparing parasitological vs serological determination of Schistosoma haematobium infection prevalence in preschool and primary school-aged children: implications for control programmes.
To combat schistosomiasis, the World Health Organization (WHO) recommends that infection levels are determined prior to designing and implementing control programmes, as the treatment regimens depend on the population infection prevalence. However, the sensitivity of the parasitological infection diagnostic method is less reliable when infection levels are low. The aim of this study was to compare levels of Schistosoma haematobium infection obtained by the parasitological method vs serological technique. Infection levels in preschool and primary school-aged children and their implications for control programmes were also investigated. Infection prevalence based on serology was significantly higher compared with that based on parasitology for both age groups. The difference between infection levels obtained using the two methods increased with age. Consequentially, in line with the WHO guidelines, the serological method suggested a more frequent treatment regimen for this population compared with that implied by the parasitological method. These findings highlighted the presence of infection in children aged ⩽5 years, further reiterating the need for their inclusion in control programmes. Furthermore, this study demonstrated the importance of using sensitive diagnostic methods as this has implications on the required intervention controls for the population.


Light sheet microscopy reveals more gradual light attenuation in light-green versus dark-green soybean leaves.
Light wavelengths preferentially absorbed by chlorophyll (chl) often display steep absorption gradients. This over-saturates photosynthesis in upper chloroplasts and deprives lower chloroplasts of blue and red light. Reducing chl content could create a more even leaf light distribution and thereby increase leaf light-use efficiency and overall canopy photosynthesis. This was tested on soybean cultivar 'Clark' (WT) and a near-isogenic chl b deficient mutant, Y11y11, grown in controlled environment chambers and in the field. Light attenuation was quantified using a novel approach involving light sheet microscopy. Leaf adaxial and abaxial surfaces were illuminated separately with blue, red, and green wavelengths, and chl fluorescence was detected orthogonally to the illumination plane. Relative fluorescence was significantly greater in deeper layers of the Y11y11 mesophyll than in WT, with the greatest differences in blue, then red, and finally green light when illuminated from the adaxial surface. Modeled relative photosynthesis based on chlorophyll profiles and Beer's Law predicted less steep gradients in mutant relative photosynthesis rates compared to WT. Although photosynthetic light-use efficiency was greater in the field-grown mutant with ~50% lower chl, light-use efficiency was lower in the mutant when grown in chambers where chl was ~80% reduced. This difference is probably due to pleiotropic effects of the mutation that accompany very severe reductions in chlorophyll and may warrant further testing in other low-chl lines.


Development and characterization of dilutable self-microemulsifying premicroemulsion systems (SMEPMS) as templates for preparation of nanosized particulates.
The utilization of self-microemulsifying premicroemulsion systems (SMEPMS) as templates for preparing poorly water-soluble compounds in the nanosized range represents a promising strategy. Fenofibrate was formulated with n-butyl L-lactate, Tween 80, and a number of cosurfactants (ethanol, 1-propanol, and PEG 600), diluted with the water phase (either water or saccharide solution) and then subjected to a freeze-drying (FD) process to obtain SMEPMS nanosized particulates. Results demonstrated that the particle size after resuspension of these FD SMEPMS nanosized particulates in water was too large, so the addition of saccharide solutions (lactose, mannitol, glucose, sucrose, and trehalose) as the solid carrier to prevent particles from aggregating seemed to be necessary and workable due to steric hindrance and repulsion. However, instability of these resuspended FD nanosized particulates after 30-90 minutes still occurred, and the addition of 0.5% sodium lauryl sulfate in the resuspending medium was able to retard the aggregation and maintain the particle size within the nano-range. Evaluation by scanning electron microscopy and X-ray powder diffraction also confirmed the results. It was concluded that using an SMEPMS formulation with PEG 600 as the cosurfactant, and in the presence of a suitable saccharide as an anticaking agent and FD process were able to produce fenofibrate nanoparticles.


Synthesis of All-Carbon Disubstituted Bicyclo[1.1.1]pentanes by Iron-Catalyzed Kumada Cross-Coupling.
1,3-Disubstituted bicyclo[1.1.1]pentanes (BCPs) are important motifs in drug design as surrogates for p-substituted arenes and alkynes. Access to all-carbon disubstituted BCPs via cross-coupling has to date been limited to use of the BCP as the organometallic component, which restricts scope due to the harsh conditions typically required for the synthesis of metallated BCPs. Here we report a general method to access 1,3-C-disubstituted BCPs from 1-iodo-bicyclo[1.1.1]pentanes (iodo-BCPs) by direct iron-catalyzed cross-coupling with aryl and heteroaryl Grignard reagents. This chemistry represents the first general use of iodo-BCPs as electrophiles in cross-coupling, and the first Kumada coupling of tertiary iodides. Benefiting from short reaction times, mild conditions, and broad scope of the coupling partners, it enables the synthesis of a wide range of 1,3-C-disubstituted BCPs including various drug analogues.


Immediate and delayed potentiating effects of tumor necrosis factor-α on TRPV1 sensitivity of rat vagal pulmonary sensory neurons.
We studied acute effects of tumor necrosis factor-α (TNFα) on the sensitivity of isolated rat vagal pulmonary sensory neurons. Our results showed the following. First, a brief pretreatment with a low dose of TNFα (1.44 nM, 9 min) enhanced the sensitivity of transient receptor potential vanilloid type 1 (TRPV1) receptors in these neurons in two distinct phases: the inward current evoked by capsaicin was amplified (Δ = 247%) immediately following the TNFα pretreatment, which gradually declined toward control and then increased again reaching another peak (Δ = 384%) after 60-90 min. Second, the immediate phase of this potentiating effect of TNFα was completely abolished by a pretreatment with a selective cyclooxygenase-2 (COX-2) inhibitor, NS-398, whereas the delayed potentiation was only partially attenuated. Third, in sharp contrast, TNFα did not generate any potentiating effect on the responses to non-TRPV1 chemical activators of these neurons. Fourth, the selectivity of the TNFα action on TRPV1 was further illustrated by the responses to acid (pH 6.0); TNFα did not affect the rapid transient current mediated by acid-sensing ion channels but significantly augmented the slow sustained current mediated by TRPV1 in the same neurons. Fifth, in anesthetized rats, a similar pattern of acute sensitizing effects of TNFα on pulmonary C-fiber afferents and the involvement of COX-2 were also clearly shown. In conclusion, a brief pretreatment with TNFα induced both immediate and delayed potentiating effects on the TRPV1 sensitivity in pulmonary sensory neurons, and the production of COX-2 arachidonic acid metabolites plays a major role in the immediate sensitizing effect of TNFα.


Tisagenlecleucel cellular kinetics, dose, and immunogenicity in relation to clinical factors in relapsed/refractory DLBCL.
The anti-CD19 chimeric antigen receptor (CAR)-T cell therapy tisagenlecleucel was evaluated in the global, phase 2 JULIET study in adult patients with relapsed/refractory diffuse large B-cell lymphoma (DLBCL). We correlated tisagenlecleucel cellular kinetics with clinical/product parameters in 111 patients treated in JULIET. Tisagenlecleucel persistence in responders and nonresponders, respectively, was demonstrated for 554 and 400 days maximum by flow cytometry and for 693 and 374 days maximum by quantitative polymerase chain reaction (qPCR). No relationships were identified between cellular kinetics (qPCR) and product characteristics, intrinsic/extrinsic factors, dose, or immunogenicity. Most patients with 3-month response had detectable transgene at time of response and continued persistence for ≥6 months. Expansion (maximal expansion of transgene/CAR-positive T-cell levels in vivo postinfusion [Cmax]) was potentially associated with response duration but this did not reach statistical significance (hazard ratio for a twofold increase in Cmax, 0.79; 95% confidence interval, 0.61-1.01). Tisagenlecleucel expansion was associated with cytokine-release syndrome (CRS) severity and tocilizumab use; no relationships were observed with neurologic events. Transgene levels were associated with B-cell levels. Dose was associated with CRS severity, but this was not statistically significant after adjusting for baseline tumor burden. In contrast to the results from B-cell precursor acute lymphoblastic leukemia (B-ALL) and chronic lymphocytic leukemia, similar exposure was observed in DLBCL in this study regardless of response and expansion was lower in DLBCL than B-ALL, likely from differences in cancer location and/or T-cell intrinsic factors. Relationships between expansion and CRS severity, and lack of relationships between dose and exposure, were similar between DLBCL and B-ALL. Tisagenlecleucel cellular kinetics in adult relapsed/refractory DLBCL improve current understanding of in vivo expansion and its relationships with safety/efficacy endpoints. This trial was registered at www.clinicaltrials.gov as #NCT02445248.


Chairside polishing of heat-cured acrylic resin: an SEM and EDA study.
This study investigated the surfaces produced on two commonly used heat-cured acrylic resins by two chairside polishing systems compared to conventional lathe polishing. It used energy dispersive analysis (EDA) and scanning electron microscopy (SEM) to investigate the abrasive materials bonded in the silicone points that are marketed for chairside polishing of acrylic resin. Using the three polishing systems (conventional as control, Shofu acrylic polishing system [AcryPoint], and Kenda Queen silicone polisher), three operators polished specimens using a defined protocol. Specimens were subjected to SEM and confocal microscopy. EDA was undertaken on the silicone polishing points. Conventional polishing produced the superior surface, followed by the surfaces produced by the Shofu system and then Kenda Queen. However, both chairside polishing systems produced smoother surfaces on acrylic resin than the surfaces produced by either steel or tungsten carbide burs. Both the Shofu acrylic polishing system and Kenda Queen silicone polisher contain beads of cerium, which is widely used in industry for polishing of glass. Silicone polishing points marketed for polishing of acrylic resin could be used to produce smooth surfaces that have been adjusted with burs where there is no access to a laboratory lathe.


Roles of eyes, leg proprioceptors and statocysts in the compensatory eye movements of freely walking land crabs (Cardisoma guanhumi)
The compound eyes, the canal organs of the statocysts and proprioceptors in the legs all generate compensatory eye movements in the horizontal plane in the land crab Cardisoma guanhumi. Frequency analyses of the compensatory eye reflexes elicited by each of these inputs show that visual (V) and proprioceptive (P) reflexes respond best below 0.1 Hz, while statocyst (S) reflexes only achieve a high gain above this frequency. They thus increase the range of frequencies over which compensation can occur. Eye and body movements were recorded in an arena under all possible combinations of crabs seeing or blind (V+ or V-), with or without statocysts (S+ or S-) and freely walking or passively transported on a trolley (P+ or P-). Intact crabs (V+S+P+) show good stabilisation of the eyes in space, the only movements with respect to external coordinates being saccadic resetting movements (fast phases of nystagmus). The eyes thus compensate well for body turns, but are unaffected by translatory movements of the body and turns that are not accompanied by a change in the orientation of the long axis of the body in space. In the absence of any one sense, compensation for rotation is significantly impaired, whether measured by the increase in the width of the histograms of changes in the angular positions of the eyes in space ( capdelta &phgr; E), by the mean angular velocity of the eyes (slope of regression line, mE) with respect to the angular velocity of the body (mB) or by response gain plotted against angular acceleration of body turn (a). The absence of two senses reduces the crab's ability to compensate still further, with the statocyst-only condition (V-S+P-) being little better than the condition when all three senses are absent (V-S-P-).Such multisensory control of eye compensation for body rotation is discussed both in terms of making use of every available cue for reducing retinal slip and in making available the information content of the optic flow field.


Molecular basis of ligand dissociation in β-adrenergic receptors.
The important and diverse biological functions of β-adrenergic receptors (βARs) have promoted the search for compounds to stimulate or inhibit their activity. In this regard, unraveling the molecular basis of ligand binding/unbinding events is essential to understand the pharmacological properties of these G protein-coupled receptors. In this study, we use the steered molecular dynamics simulation method to describe, in atomic detail, the unbinding process of two inverse agonists, which have been recently co-crystallized with β(1) and β(2)ARs subtypes, along four different channels. Our results indicate that this type of compounds likely accesses the orthosteric binding site of βARs from the extracellular water environment. Importantly, reconstruction of forces and energies from the simulations of the dissociation process suggests, for the first time, the presence of secondary binding sites located in the extracellular loops 2 and 3 and transmembrane helix 7, where ligands are transiently retained by electrostatic and Van der Waals interactions. Comparison of the residues that form these new transient allosteric binding sites in both βARs subtypes reveals the importance of non-conserved electrostatic interactions as well as conserved aromatic contacts in the early steps of the binding process.


The unobjectionable part of the transference.
Even those aspects of transference which initially favor the analytic process and seem to have the least connection with resistance do become integral parts of the transference neurosis and contribute massively to some of the most subtle difficulties in the process, especially in its resolution. These phenomena are, by their very appearance of rationality and cooperation, all the more difficult to bring under analytic scrutiny. They operate as resistances not only to the analysis of preoedipal conflicts, but even more effectively in the case of neurotic disorders centered on inadequate resolution of oedipal conflicts, i.e., in the so-called classical neurosis, "the case of the ideal analytic patient." I suggest, therefore, that the appearance of the "unobjectionable component" be regarded not only as a welcome manifestation of certain conflict-free psychic elements, but also as the manifest resultant of a complex web of unconscious conflicts which must be, and are capable of being, sought for and described. Further, that their analysis may be facilitated by the use of a process analogous to that employed in the analysis of dreams, particularly with respect to secondary revision. Finally, I have emphasized that these aspects of transference, which we are tempted to explain, and in effect dismiss, by reductionist references to early development, be regarded rather as the complex resultant of a prolonged historical experience, and that they be so interpreted.


The effect of Swedish and American smokeless tobacco extract on periodontal ligament fibroblasts in vitro.
Use of moist snuff is widespread in Sweden. In 2004 approximately 8oo,ooo Swedes were daily users which corresponds to 22% of the male population and 3% of the female population. The aim of the present study was to evaluate the effect of Swedish moist snuff extract on PDLfibroblast growth and hard tissue production and compare with moist snuff extract from USA. Periodontal ligament cells (PDL-cells) were obtained from 3 healthy subjects (1 female 14 years, 2 males 14 and 17 years) from the root surface of premolars extracted for orthodontic reasons. The cells were isolated from explants and grown in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum (FBS) and cultivated in 37 degrees C with 5% CO2 in air. Snuff extract in concentrations 0.3%, 1% and 3% (in DMEM with 1% FBS) was tested. Cells from each individual were tested three times, each time in triplicate. Photographs were taken at o and 24 hours with a digital camera and analysed in terms of growth and morphology. Then the cell suspension was frozen and later thawed for examination of the production of alkaline phosphatase after exposure to different snuff concentrations. This in vitro study has shown that PDL cells from 3 different subjects demonstrated a reduced number of cells at exposure to 3% of both Swedish and American snuff extract. The production of alkaline phosphatase after 2 hours was similarly reduced from cells exposed to 3% snuff extract. Further studies have to be made to understand the effect of smokeless tobacco on periodontal tissues. However, from this study can be concluded that smokeless tobacco has biological effects in terms of reduced PDL cell growth and production of alkaline phosphatase


The global metabolic transition: Regional patterns and trends of global material flows, 1950-2010.
Since the World War II, many economies have transitioned from an agrarian, biomass-based to an industrial, minerals-based metabolic regime. Since 1950, world population grew by factor 2.7 and global material consumption by factor 3.7-71 Gigatonnes per year in 2010. The expansion of the resource base required by human societies is associated with growing pressure on the environment and infringement on the habitats of other species. In order to achieve a sustainability transition, we require a better understanding of the currently ongoing metabolic transition and its potential inertia. In this article, we present a long-term global material flow dataset covering material extraction, trade, and consumption of 177 individual countries between 1950 and 2010. We trace patterns and trends in material flows for six major geographic and economic country groupings and world regions (Western Industrial, the (Former) Soviet Union and its allies, Asia, the Middle East and Northern Africa, Latin America and the Caribbean, and Sub-Saharan Africa) as well as their contribution to the emergence of a global metabolic profile during a period of rapid industrialization and globalization. Global average material use increased from 5.0 to 10.3 tons per capita and year (t/cap/a) between 1950 and 2010. Regional metabolic rates range from 4.5 t/cap/a in Sub-Saharan Africa to 14.8 t/cap/a in the Western Industrial grouping. While we can observe a stabilization of the industrial metabolic profile composed of relatively equal shares of biomass, fossil energy carriers, and construction minerals, we note differences in the degree to which other regions are gravitating toward a similar form of material use. Since 2000, Asia has overtaken the Western Industrial grouping in terms of its share in global resource use although not in terms of its per capita material consumption. We find that at a sub-global level, the roles of the world regions have changed. There are, however, no signs yet that this will lead to stabilization or even a reduction of global resource use.


Structure-function analyses of thrombomodulin by gene-targeting in mice: the cytoplasmic domain is not required for normal fetal development.
Thrombomodulin (TM) is a widely expressed glycoprotein receptor that plays a physiologically important role in maintaining normal hemostatic balance postnatally. Inactivation of the TM gene in mice results in embryonic lethality without thrombosis, suggesting that structures of TM not recognized to be involved in coagulation might be critical for normal fetal development. Therefore, the in vivo role of the cytoplasmic domain of TM was studied by using homologous recombination in ES cells to create mice that lack this region of TM (TMcyt/cyt). Cross-breeding of F1 TMwt/cyt mice (1 wild-type and 1 mutant allele) resulted in more than 300 healthy offspring with a normal Mendelian inheritance pattern of 25.7% TMwt/wt, 46.6% TMwt/cyt, and 27.7% TMcyt/cyt mice, indicating that the tail of TM is not necessary for normal fetal development. Phenotypic analyses showed that the TMcyt/cyt mice responded identically to their wild-type littermates after procoagulant, proinflammatory, and skin wound challenges. Plasma levels of plasminogen, plasminogen activator inhibitor 1 (PAI-1), and alpha2-antiplasmin were unaltered, but plasmin:alpha2-antiplasmin (PAP) levels were significantly lower in TMcyt/cyt mice than in TMwt/wt mice (0.46 +/- 0.2 and 1.99 +/- 0.1 ng/mL, respectively; P <.001). Tissue levels of TM antigen were also unaffected. However, functional levels of plasma TM in the TMcyt/cyt mice, as measured by thrombin-dependent activation of protein C, were significantly increased (P <.001). This supported the hypothesis that suppression in PAP levels may be due to augmented activation of thrombin-activatable fibrinolysis inhibitor (TAFI), with resultant inhibition of plasmin generation. In conclusion, these studies exclude the cytoplasmic domain of TM from playing a role in the early embryonic lethality of TM-null mice and support its function in regulating plasmin generation in plasma.


[Prognostic implication of iris phenotype in ischemic heart disease].
Specific features of the eye iris were studied at iridobiomicroscopy in 250 patients with verified ischemic heart disease (IHD). Criteria of its iridodiagnosis have been formulated. The detected associations of HLA-antigens A3, B14, B15, B41 and Cw3 with typical for IHD iris changes confirmed hereditary predisposition of the main iridomarks. The results of iridoscopy may appear useful for prosnostification of IHD risk. It is suggested to consider iridologic phenotype characteristics among genetic markers indicating predisposition to IHD.


Syphilis, sex and psychiatry, 1789-1925: Part 2.
Syphilis has changed the course of history, shaped the path of medicine and had more influence on psychiatry than any other illness. This paper, part two of a two-part series, investigates the historical, social and cultural aspects of the interaction of syphilis and psychiatry. By the end of the 19th century, social changes such as population growth, mass migration from Eastern Europe and technological developments led to a great rise in syphilis. By 1900, it was estimated that 5-20% of the population of Europe and the USA had, or would have, syphilis. By 1914, there were over 100,00 new cases and 3 million cases of syphilis in Great Britain. There was a constant interaction between syphilis, prostitution and sexual crime; it was the likely motivation for the Jack the Ripper murders, if not many in the next century. The idea of hereditary syphilis fitted perfectly into the theory of degeneration and coursed through psychiatry and caught the attention of Adolf Hitler, facilitating his antisemitic paranoia. Psychiatric progress passed to the German school, led by Kraepelin who did his first research into the symptoms and course of neurosyphilis. In 1906, Wasserman's serological test for syphilis showed that latent lesions could be present. Any doubt about the cause of syphilis was finally eliminated when Noguchi and Moore demonstrated the presence of treponema pallidum in paretic brains in 1913. German academic psychiatry defined psychiatric practice for the next century but malariotherapy, the first physical treatment in psychiatry, was announced by Julius Wagner-Juarreg in Vienna in 1917, bringing hope to the incurable and destroying the climate of therapeutic nihilism that haunted psychiatry. The first trial of malariotherapy in Australia was done by Reginald Ellery at Mont Park Hospital In 1927 in Melbourne. The discovery of penicillin was a caesura, ending malariotherapy and leading many to regard syphilis as a night-extinct illness, but this turned out to be an illusion. Syphilis is returning in new forms in tandem with the AIDS epidemic. Written-off endlessly by its obituarists, syphilis abides.


Mechanism of action of cytochalasin: evidence that it binds to actin filament ends.
To test the idea that cytochalasin retards actin assembly by binding to filament ends, we have designed a new assay for cytochalasin binding in which the number of filament ends can be varied independently of the total actin concentration. Actin is reacted with polylysine-coated polystyrene beads to make filament ends (Brown and Spudich, 1979, J. Cell Biol. 80:499-504) and then reacted with [3H]cytochalasin B. We have found that cytochalasin B binds to beads in the presence of actin, and that the number of cytochalasin B binding sites can be varied as a function of the number of filament ends independent of the total actin concentration by varying the bead concentration.


Numerical abstraction in young domestic chicks (Gallus gallus).
In a variety of circumstances animals can represent numerical values per se, although it is unclear how salient numbers are relative to non-numerical properties. The question is then: are numbers intrinsically distinguished or are they processed as a last resort only when no other properties differentiate stimuli? The last resort hypothesis is supported by findings pertaining to animal studies characterized by extensive training procedures. Animals may, nevertheless, spontaneously and routinely discriminate numerical attributes in their natural habitat, but data available on spontaneous numerical competence usually emerge from studies not disentangling numerical from quantitative cues. In the study being outlined here, we tested animals' discrimination of a large number of elements utilizing a paradigm that did not require any training procedures. During rearing, newborn chicks were presented with two stimuli, each characterized by a different number of heterogeneous (for colour, size and shape) elements and food was found in proximity of one of the two stimuli. At testing 3 day-old chicks were presented with stimuli depicting novel elements (for colour, size and shape) representing either the numerosity associated or not associated with food. The chicks approached the number associated with food in the 5vs.10 and 10vs.20 comparisons both when quantitative cues were unavailable (stimuli were of random sizes) or being controlled. The findings emerging from the study support the hypothesis that numbers are salient information promptly processed even by very young animals.


Development of a standard operating procedure and checklist for rapid sequence induction in the critically ill.
Rapid sequence induction (RSI) of critically ill patients outside of theatres is associated with a higher risk of hypoxia, cardiovascular collapse and death. In the prehospital and military environments, there is an increasing awareness of the benefits of standardised practice and checklists. We conducted a non-systematic review of literature pertaining to key components of RSI preparation and management. A standard operating procedure (SOP) for in-hospital RSI was developed based on this and experience from large teaching hospital anaesthesia and critical care departments. The SOP consists of a RSI equipment set-up sheet, pre-RSI checklist and failed airway algorithm. The SOP should improve RSI preparation, crew resource management and first pass intubation success while minimising adverse events. Based on the presented literature, we believe the evidence is sufficient to recommend adoption of the core components in the suggested SOP. This standardised approach to RSI in the critically ill may reduce the current high incidence of adverse events and hopefully improve patient outcomes.


Preferential glutathione conjugation of a reverse diol epoxide compared with a bay region diol epoxide of benzo[a]pyrene in human hepatocytes.
Many studies have examined the relationship between polymorphisms in glutathione S-transferase genes and cancer in people exposed to polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyrene (BaP), but the results to date have been modest. Missing from these studies has been an exploration of the formation of the appropriate glutathione conjugates in humans. We incubated human hepatocytes from 10 donors with racemic anti-BaP-7,8-diol-9,10-epoxide (BPDE), believed to be a major ultimate carcinogen of BaP, or with the noncarcinogenic reverse diol epoxide, racemic anti-BaP-9,10-diol-7,8-epoxide (rev-BPDE). Incubations were carried out for 12 or 24 h. We used high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry-selected reaction monitoring at m/z 464 --> m/z 317 to analyze the incubation mixtures for the mercapturic acid products that would result from glutathione conjugation. The standard mercapturic acids were synthesized by reaction of BPDE or rev-BPDE with N-acetylcysteine. We obtained convincing evidence in human hepatocytes for mercapturic acid formation from rev-BPDE in all 10 samples, in amounts up to 17 pmol/ml. However, we could detect mercapturic acids from BPDE in only 1 of 10 samples (0.05 pmol/ml). Taken together with our similar previous results of analyses of phenanthrene metabolites in human hepatocytes and human urine, the results of this study indicate that conjugation of BPDE with glutathione is a minor pathway in humans, indicating that glutathione S-transferase genotyping is not an effective method for assessing risk of PAH-induced cancer in humans, at least with respect to the diol epoxide pathway of PAH carcinogenesis.


Comparison of 2-D and 3-D estimates of placental volume in early pregnancy.
Ultrasound estimation of placental volume (PlaV) between 11 and 13 wk has been proposed as part of a screening test for small-for-gestational-age babies. A semi-automated 3-D technique, validated against the gold standard of manual delineation, has been found at this stage of gestation to predict small-for-gestational-age at term. Recently, when used in the third trimester, an estimate obtained using a 2-D technique was found to correlate with placental weight at delivery. Given its greater simplicity, the 2-D technique might be more useful as part of an early screening test. We investigated if the two techniques produced similar results when used in the first trimester. The correlation between PlaV values calculated by the two different techniques was assessed in 139 first-trimester placentas. The agreement on PlaV and derived "standardized placental volume," a dimensionless index correcting for gestational age, was explored with the Mann-Whitney test and Bland-Altman plots. Placentas were categorized into five different shape subtypes, and a subgroup analysis was performed. Agreement was poor for both PlaV and standardized PlaV (p < 0.001 and p < 0.001), with the 2-D technique yielding larger estimates for both indices compared with the 3-D method. The mean difference in standardized PlaV values between the two methods was 0.007 (95% confidence interval: 0.006-0.009). The best agreement was found for regular rectangle-shaped placentas (p = 0.438 and p = 0.408). The poor correlation between the 2-D and 3-D techniques may result from the heterogeneity of placental morphology at this stage of gestation. In early gestation, the simpler 2-D estimates of PlaV do not correlate strongly with those obtained with the validated 3-D technique.


Attributable mortality of central line associated bloodstream infection: systematic review and meta-analysis.
To identify the attributable mortality of central line associated blood stream infections (CLABSI) through meta-analysis. Meta-analysis of case control and cohort studies, matched and unmatched, that reported on mortality of patients with and without CLABSI was performed. MEDLINE, CENTRAL, CINAHL were searched. Non-interventional studies of all languages that reported mortality in patients with CLABSI were included. Data were extracted on patient population, study setting, design, diagnostic criteria for CLABSI, and mortality. Results from studies comparing mortality due to CLABSI were pooled using a random effects model with assessment of heterogeneity. Heterogeneity of studies was assessed with an I (2) statistic and a funnel plot was generated to assess for publication bias. Eighteen studies were included with 1,976 CLABSI cases. Of the included studies, 17 took place in intensive care unit settings, most involved a mixed population of medical and surgical patients, and ten were matched using an illness severity index. Our findings show an odds ratio of in hospital death associated with CLABSI as 2.75 (CI 1.86-4.07) and 1.51 (CI 1.08-2.09) in the subgroup of the ten matched studies. Those studies where greater than 30 % of CLABSI were attributed to coagulase-negative Staphylococcus had an odds ratio of death of 1.64 (95 % CI 1.02-2.65) compared with 4.71 (95 % CI 1.54-14.39). CLABSI is associated with a significantly increased risk of death supporting the use of extensive efforts to reduce these infections.


Interventions integrating health and academic interventions to prevent substance use and violence: a systematic review and synthesis of process evaluations.
Within increasingly constrained school timetables, interventions that integrate academic and health education to reduce substance use and violence may hold promise as a category of intervention that can positively affect both academic and health outcomes. There are no current systematic reviews exploring the effectiveness of such interventions or factors that affect their implementation. A total of 19 bibliographic databases and 32 websites were searched. References were also extracted from the reference lists of included studies, and experts and authors were contacted to identify relevant studies. We included reports with no restrictions on language or date. References were screened on title/abstract and those not thus excluded were screened on full report. Data extraction and appraisal followed the Critical Appraisal Skills Programme, Evidence for Policy and Practice Information and Co-ordinating Centre and Cochrane tools. Extracted process data were qualitatively meta-synthesised for common themes. Seventy-eight thousand four hundred fifty-one unique references were identified, and 62 reports were included. A total of 16 reports (reporting on 15 studies of 12 interventions) evaluated process. Key facilitators of integrated academic and health curricula were supportive senior management and alignment of the intervention with school ethos; a positive teaching environment, including positive perceptions around the ability to be flexible in the adaptation and delivery of integrated academic and health curricula; positive pre-existing student and teacher attitudes towards intervention content; and parental support of interventions, largely through reinforcement of messaging at home. Important barriers were over-burdened teachers, with little time to learn and implement integrated curricula. Several useful facilitating and inhibiting factors linked to the implementation of interventions that integrate academic and health education for reduced substance use and/or violence were identified, providing tentative but insightful evidence of context-specific issues that may impact intervention success. However, overall, there is still a considerable gap in our understanding of how to achieve the successful implementation of these interventions.


Secretory apparatus assessed by analysis of pancreatic secretory stress protein expression in a rat model of chronic pancreatitis.
Secretory stress proteins (SSP) are a family of proteins including isoforms of pancreatitis-associated protein (PAP) and pancreatic stone protein (PSP/reg). In vitro exposure to trypsin results in the formation of insoluble fibrillar structures. SSP are constitutively secreted into pancreatic juice at low levels. The WBN/Kob rat is a model for chronic pancreatitis, displaying focal inflammation, destruction of the parenchyma and changes in the architecture of the acinar cell; the synthesis and secretion of SSP are also increased. We have investigated the secretory apparatus by SSP immunohistochemistry at the light- and electron-microscopical (EM) levels. Immunocytochemistry of PSP/reg in Wistar control rats reveals low levels, with individual acinar cells exhibiting high immunoreactivity in zymogen granules. PAP is not detectable. In the WBN/Kob rat, PSP/reg and PAP immunoreactivity is markedly increased. Double immunofluorescence for PSP/reg and PAP I or II demonstrates that these proteins colocalize to the same cell. Acinar cells change their secretory architecture by fusion of zymogen granules and elongation of the fused organelles. The immunogold technique has demonstrated an increase of SSP in zymogen granules in WBN/Kob rats. PSP/reg-positive zymogen granules fuse to form elongated structures with fibrillar contents. An extensive PSP/reg-positive fibrillar network is established in the cytosol. Extracellular fibrils have been observed in several ductules. Thus, SSP-derived fibrils form concomitantly with acinar damage in the WBN/Kob rat. Based on the known tryptic cleavage site of SSP, the in vivo generation of fibrils is presumably the result of premature trypsin activation.


NEUROBID--an EU-funded project to study the developing brain barriers.
Brain diseases are one of the most prevalent groups of diseases in Europe with estimated annual costs amounting to euro386 billion. Data collected by the WHO suggest that brain diseases are responsible for 35% of Europe's total disease burden. In the treatment of neurological disease, the blood brain barrier (BBB) still represents an obstacle for the delivery of drugs to the brain and thus a major challenge for the development of therapeutic regimens. Understanding the molecular basis and functioning of the BBB in health and disease, including transport mechanisms across the BBB, therefore holds significant potential for future strategies to prevent and ameliorate neurological disease. Recent research indicates that some neurological disorders have a developmental etiologic component. The major goal of the NEUROBID project is thus to understand the molecular mechanisms and function of the BBB in health and disease both in the developing brain and the adult central nervous system. With an interdisciplinary consortium from the fields of developmental neurobiology and BBB research, NEUROBID aims to (i) understand the involvement of normal and disturbed BBB function in normal and abnormal brain development and (ii) to develop novel strategies for drug delivery to the brain. Unique transport mechanisms across the BBB will be used to target potential therapeutic macromolecular and cellular agents specifically to the brain barriers and transport them into the brain. The main target disorders of NEUROBID are non-inherited neurodevelopmental disorders arising from perinatal adverse exposure, such as cerebral palsy, and classic adult neurological disorders such as multiple sclerosis and stroke. In the long term, NEUROBID hopes to pave the way for new treatment strategies and thus reduce the economic and social burden of neurological disease.


Semiparametric estimation of covariance matrices for longitudinal data.
Estimation of longitudinal data covariance structure poses significant challenges because the data are usually collected at irregular time points. A viable semiparametric model for covariance matrices was proposed in Fan, Huang and Li (2007) that allows one to estimate the variance function nonparametrically and to estimate the correlation function parametrically via aggregating information from irregular and sparse data points within each subject. However, the asymptotic properties of their quasi-maximum likelihood estimator (QMLE) of parameters in the covariance model are largely unknown. In the current work, we address this problem in the context of more general models for the conditional mean function including parametric, nonparametric, or semi-parametric. We also consider the possibility of rough mean regression function and introduce the difference-based method to reduce biases in the context of varying-coefficient partially linear mean regression models. This provides a more robust estimator of the covariance function under a wider range of situations. Under some technical conditions, consistency and asymptotic normality are obtained for the QMLE of the parameters in the correlation function. Simulation studies and a real data example are used to illustrate the proposed approach.


Pleiotropic role for MYCN in medulloblastoma.
Medulloblastoma (MB) is the most common malignant brain tumor of childhood. Sonic Hedgehog (SHH) signaling drives a minority of MB, correlating with desmoplastic pathology and favorable outcome. The majority, however, arises independently of SHH and displays classic or large cell anaplastic (LCA) pathology and poor prognosis. To identify common signaling abnormalities, we profiled mRNA, demonstrating misexpression of MYCN in the majority of human MB and negligible expression in normal cerebella. We clarified a role in pathogenesis by targeting MYCN (and luciferase) to cerebella of transgenic mice. MYCN-driven MB showed either classic or LCA pathologies, with Shh signaling activated in approximately 5% of tumors, demonstrating that MYCN can drive MB independently of Shh. MB arose at high penetrance, consistent with a role for MYCN in initiation. Tumor burden correlated with bioluminescence, with rare metastatic spread to the leptomeninges, suggesting roles for MYCN in both progression and metastasis. Transient pharmacological down-regulation of MYCN led to both clearance and senescence of tumor cells, and improved survival. Targeted expression of MYCN thus contributes to initiation, progression, and maintenance of MB, suggesting a central role for MYCN in pathogenesis.


Safety and effectiveness of hormonal treatment versus hormonal treatment with vigabatrin for infantile spasms (ICISS): a randomised, multicentre, open-label trial.
Infantile spasms constitutes a severe infantile epilepsy syndrome that is difficult to treat and has a high morbidity. Hormonal therapies or vigabatrin are the most commonly used treatments. We aimed to assess whether combining the treatments would be more effective than hormonal therapy alone. In this multicentre, open-label randomised trial, 102 hospitals (Australia [three], Germany [11], New Zealand [two], Switzerland [three], and the UK [83]) enrolled infants who had a clinical diagnosis of infantile spasms and a hypsarrhythmic (or similar) EEG no more than 7 days before enrolment. Participants were randomly assigned (1:1) by a secure website to receive hormonal therapy with vigabatrin or hormonal therapy alone. If parents consented, there was an additional randomisation (1:1) of type of hormonal therapy used (prednisolone or tetracosactide depot). Block randomisation was stratified for hormonal treatment and risk of developmental impairment. Parents and clinicians were not masked to therapy, but investigators assessing electro-clinical outcome were masked to treatment allocation. Minimum doses were prednisolone 10 mg four times a day or intramuscular tetracosactide depot 0·5 mg (40 IU) on alternate days with or without vigabatrin 100 mg/kg per day. The primary outcome was cessation of spasms, which was defined as no witnessed spasms on and between day 14 and day 42 from trial entry, as recorded by parents and carers in a seizure diary. Analysis was by intention to treat. The trial is registered with The International Standard Randomised Controlled Trial Number (ISRCTN), number 54363174, and the European Union Drug Regulating Authorities Clinical Trials (EUDRACT), number 2006-000788-27. Between March 7, 2007, and May 22, 2014, 766 infants were screened and, of those, 377 were randomly assigned to hormonal therapy with vigabatrin (186) or hormonal therapy alone (191). All 377 infants were assessed for the primary outcome. Between days 14 and 42 inclusive no spasms were witnessed in 133 (72%) of 186 patients on hormonal therapy with vigabatrin compared with 108 (57%) of 191 patients on hormonal therapy alone (difference 15·0%, 95% CI 5·1-24·9, p=0·002). Serious adverse reactions necessitating hospitalisation occurred in 33 infants (16 on hormonal therapy alone and 17 on hormonal therapy with vigabatrin). The most common serious adverse reaction was infection occurring in five infants on hormonal therapy alone and four on hormonal therapy with vigabatrin. There were no deaths attributable to treatment. Hormonal therapy with vigabatrin is significantly more effective at stopping infantile spasms than hormonal therapy alone. The 4 week period of spasm cessation required to achieve a primary clinical response to treatment suggests that the effect seen might be sustained, but this needs to be confirmed at the 18 month follow-up. The Castang Foundation, Bath Unit for Research in Paediatrics, National Institute of Health Research, the Royal United Hospitals Bath NHS Foundation Trust, the BRONNER-BENDUNG Stifung/Gernsbach, and University Children's Hospital Zurich.


Tuning the Kondo resonance in two-dimensional lattices of cerium molecular complexes.
Cerium intermetallics have raised a lot of interest for the past forty years thanks to their very unusual and interesting electronic and magnetic properties. This can be explained by the peculiar electronic configuration of Ce (4f1) that allows different oxidation states leading to singular behavior such as quantum phase transitions, heavy-fermion behavior and the Kondo effect. In this work, we used a mixed-valence molecular analogue to study the Kondo effect down to the atomic scale by means of scanning tunneling microscopy/spectroscopy (STM/STS) for which new many-body effects are expected to emerge due to reduced dimensionality and specific chemical environment of the 4f-ion. For that purpose, double-decker molecular complexes hosting a Ce ion were synthesized and adsorbed onto Ag and Cu (111) surfaces forming two-dimensional lattices. As a result, we observed a zero-bias conductance resonance on Ag only indicative of a Kondo effect arising from the coupling between a molecular spin and the conducting electrons of the metallic surface. The emergence of the Kondo effect is discussed in terms of intermolecular and molecule/substrate interactions. This work expands the little knowledge to date on the structural and related electronic properties of Ce-based molecular systems on surfaces. In particular, it shows that Ce-based double deckers are good platforms to obtain insight into 4f-induced many-body effects down to the nanometer scale and in two-dimensional lattices. Moreover, this outcome has a strong impact for future applications of molecular devices in which both metals are commonly used as electrical contacts.


Preoperative education on postoperative delirium, anxiety, and knowledge in pulmonary thromboendarterectomy patients.
Delirium is the most common postoperative psychiatric condition in intensive care settings and can lead to increased complications and costs. To evaluate the impact of multifaceted preoperative patient education on postoperative delirium, anxiety, and knowledge and to explore predictors of postoperative delirium, days of mechanical ventilation, and days in the intensive care unit (ICU) in patients undergoing pulmonary thromboendarterectomy. A prospective, randomized controlled trial was conducted on consented patients from October 2011 to April 2013. Patients were randomized in a 1 to 1 ratio to receive either an individualized 45-minute multifaceted preoperative education (experimental group, n = 63) or standard education (control group, n = 66). Participants completed the State-Trait Anxiety Inventory and Knowledge Test before and after the education. Data on incidence of delirium, days of mechanical ventilation, ICU days, and cardiopulmonary parameters were collected. The experimental group had significantly more knowledge about postoperative care (P< .001) and fewer days of mechanical ventilation (P = .04) than the control group. The 2 groups did not differ significantly in anxiety, incidence of delirium, or ICU days. In exploratory multivariate analyses, hearing impairment was a positive predictor for days of delirium (P = .009), days of mechanical ventilation (P< .001), and ICU days (P= .049), whereas the posttest knowledge was a negative predictor for days of mechanical ventilation (P = .02). The patient education appeared to be effective in improving knowledge and reducing days of mechanical ventilation. Hearing impairment was an unexpected predictor of adverse outcomes for patients but may be amenable to nursing intervention.


Quantum coherent energy transfer over varying pathways in single light-harvesting complexes.
The initial steps of photosynthesis comprise the absorption of sunlight by pigment-protein antenna complexes followed by rapid and highly efficient funneling of excitation energy to a reaction center. In these transport processes, signatures of unexpectedly long-lived coherences have emerged in two-dimensional ensemble spectra of various light-harvesting complexes. Here, we demonstrate ultrafast quantum coherent energy transfer within individual antenna complexes of a purple bacterium under physiological conditions. We find that quantum coherences between electronically coupled energy eigenstates persist at least 400 femtoseconds and that distinct energy-transfer pathways that change with time can be identified in each complex. Our data suggest that long-lived quantum coherence renders energy transfer in photosynthetic systems robust in the presence of disorder, which is a prerequisite for efficient light harvesting.


Many LINE1 elements contribute to the transcriptome of human somatic cells.
While LINE1 (L1) retroelements comprise nearly 20% of the human genome, the majority are thought to have been rendered transcriptionally inactive, due to either mutation or epigenetic suppression. How many L1 elements 'escape' these forms of repression and contribute to the transcriptome of human somatic cells? We have cloned out expressed sequence tags corresponding to the 5' and 3' flanks of L1 elements in order to characterize the population of elements that are being actively transcribed. We also examined expression of a select number of elements in different individuals. We isolated expressed sequence tags from human lymphoblastoid cell lines corresponding to 692 distinct L1 element sites, including 410 full-length elements. Four of the expression tagged sites corresponding to full-length elements from the human specific L1Hs subfamily were examined in European-American individuals and found to be differentially expressed in different family members. A large number of different L1 element sites are expressed in human somatic tissues, and this expression varies among different individuals. Paradoxically, few elements were tagged at high frequency, indicating that the majority of expressed L1s are transcribed at low levels. Based on our preliminary expression studies of a limited number of elements in a single family, we predict a significant degree of inter-individual transcript-level polymorphism in this class of sequence.


Synthesis, structure and antimicrobial property of green composites from cellulose, wool, hair and chicken feather.
Novel composites between cellulose (CEL) and keratin (KER) from three different sources (wool, hair and chicken feather) were successfully synthesized in a simple one-step process in which butylmethylimidazolium chloride (BMIm(+)Cl(-)), an ionic liquid, was used as the sole solvent. The method is green and recyclable because [BMIm(+)Cl(-)] used was recovered for reuse. Spectroscopy (FTIR, XRD) and imaging (SEM) results confirm that CEL and KER remain chemically intact and homogeneously distributed in the composites. KER retains some of its secondary structure in the composites. Interestingly, the minor differences in the structure of KER in wool, hair and feather produced pronounced differences in the conformation of their corresponding composites with wool has the highest α-helix content and feather has the lowest content. These results correlate well with mechanical and antimicrobial properties of the composites. Specifically, adding CEL into KER substantially improves mechanical strength of [CEL+KER] composites made from all three different sources, wool, hair and chicken feathers i.e., [CEL+wool], [CEL+hair] and [CEL+feather]. Since mechanical strength is due to CEL, and CEL has only random structure, [CEL+feather] has, expectedly, the strongest mechanical property because feather has the lowest content of α-helix. Conversely, [CEL+wool] composite has the weakest mechanical strength because wool has the highest α-helix content. All three composites exhibit antibacterial activity against methicillin resistant Staphylococcus aureus (MRSA). The antibacterial property is due not to CEL but to the protein and strongly depends on the type of the keratin, namely, the bactericidal effect is strongest for feather and weakest for wool. These results together with our previous finding that [CEL+KER] composites can control release of drug such as ciprofloxacin clearly indicate that these composites can potentially be used as wound dressing.


Endoplasmic reticulum transmembrane protein TMTC3 contributes to O-mannosylation of E-cadherin, cellular adherence, and embryonic gastrulation.
Protein glycosylation plays essential roles in protein structure, stability, and activity such as cell adhesion. The cadherin superfamily of adhesion molecules carry O-linked mannose glycans at conserved sites and it was recently demonstrated that the transmembrane and tetratricopeptide repeat-containing proteins 1-4 (TMTC1-4) gene products contribute to the addition of these O-linked mannoses. Here, biochemical, cell biological, and organismal analysis was used to determine that TMTC3 supports the O-mannosylation of E-cadherin, cellular adhesion, and embryonic gastrulation. Using genetically engineered cells lacking all four TMTC genes, overexpression of TMTC3 rescued O-linked glycosylation of E-cadherin and cell adherence. The knockdown of the Tmtcs in Xenopus laevis embryos caused a delay in gastrulation that was rescued by the addition of human TMTC3. Mutations in TMTC3 have been linked to neuronal cell migration diseases including Cobblestone lissencephaly. Analysis of TMTC3 mutations associated with Cobblestone lissencephaly found that three of the variants exhibit reduced stability and missence mutations were unable to complement TMTC3 rescue of gastrulation in Xenopus embryo development. Our study demonstrates that TMTC3 regulates O-linked glycosylation and cadherin-mediated adherence, providing insight into its effect on cellular adherence and migration, as well the basis of TMTC3-associated Cobblestone lissencephaly.


Evaluation of the impact of the 2012 Rhode Island health care worker influenza vaccination regulations: implementation process and vaccination coverage.
In October 2012, the Rhode Island Department of Health (HEALTH) amended its health care worker (HCW) vaccination regulations to require all HCWs to receive annual influenza vaccination or wear a surgical mask during direct patient contact when influenza is widespread. Unvaccinated HCWs failing to wear a mask are subject to a fine and disciplinary action. To describe the implementation of the 2012 Rhode Island HCW influenza vaccination regulations and examine their impact on vaccination coverage. Two data sources were used: (1) a survey of all health care facilities subject to the HCW regulations and (2) HCW influenza vaccination coverage data reported to HEALTH by health care facilities. Descriptive statistics and paired t tests were performed using SAS Release 9.2. For the 2012-2013 influenza season, 271 inpatient and outpatient health care facilities in Rhode Island were subject to the HCW regulations. Increase in HCW influenza vaccination coverage. Of the 271 facilities, 117 facilities completed the survey (43.2%) and 160 facilities reported vaccination data to HEALTH (59.0%). Between the 2011-2012 and 2012-2013 influenza seasons, the proportion of facilities having a masking policy, as required by the revised regulations, increased from 9.4% to 94.0% (P < .001). However, the proportion of facilities implementing Advisory Committee on Immunization Practices-recommended strategies to promote HCW influenza vaccination did not increase. The majority of facilities perceived benefits to collecting HCW influenza vaccination data, including strengthening infection prevention efforts (83.2%) and improving patient and coworker safety (75.2%). Concurrent with the new regulations, influenza vaccination coverage among employee HCWs in Rhode Island increased from 69.7% in the 2011-2012 influenza season to 87.2% in the 2012-2013 season. Rhode Island's experience demonstrates that statewide HCW influenza vaccination requirements incorporating mask wearing and moderate penalties for noncompliance can be effective in improving influenza vaccination coverage among HCWs.


Degradation of the viral strand of phiX174 parental replicative-form DNA in a rep- host.
A progressive degradation of the parental viral strand label is observed upon infection of a Rep- mutant of Escherichia coli by 32P-labeled phiX174. Very little parental label remains in the RF (replicative form) by 47 min after infection. Concomitant with this degradation, replicative intermediates are formed which sediment at 21s, the rate of RF I (supercoiled-closed circular DNA), in a neutral sucrose gradient but which denature and sediment in alkaline gradients as single strands of unit size and larger. These denaturable 21s replicative intermediates have been shown previously to be RF molecules containing an elongated viral strand. Addition of chloramphenicol at 7 min after infection at 30 mug/ml, a concentration sufficient to block RF leads to SS (single strand) synthesis but not RF leads to RF synthesis in a wild-type host cell, reduced the amount of viral strand elongation but did not prevent viral strand degradation. The addition of chloramphenicol at 150 mug/ml at 7 min after infection totally prevents both the degradation of the parental label and the formation of the replicative intermediates with elongated tails. We infer that degradation of the viral strand requires the gene A-mediated nicking of the viral strand but not the concomitant elongation of the viral strand.


Lamotrigine-associated rash: to rechallenge or not to rechallenge?
The major burden of illness in bipolar disorder (BD) is in the depressive pole. Lamotrigine has been shown to be useful in the long-term prophylaxis of depressive episodes in BD. Current guidelines recommend discontinuing lamotrigine in patients who develop rash. Our objective in this paper is to review literature to identify possible predictors of serious vs. benign rash that might help guide clinical decision-making and recommend titration strategy for re-introduction of lamotrigine, if indicated. We performed a search of the literature between 1966 and July 2008 to investigate the phenomenon of lamotrigine-induced rash and rechallenge procedures. The search identified six reports, and we were able to identify another case series from reviewing the bibliography of all of the above papers. We reviewed all the papers of lamotrigine rash rechallenge that resulted from the literature search. These papers describe 44 cases of lamotrigine rechallenge. Currently, there are 39 reported cases in the literature of successful lamotrigine rechallenge after a rash and five cases with rash recurrence. There are some characteristics of the rash that can help identify serious cases from benign ones. In addition, very slow titration of lamotrigine is crucial to the reduction of rash recurrence rate. Several cases that develop benign rash on lamotrigine can be rechallenged without adverse consequences. We believe that lamotrigine rechallenge in bipolar depression is an under-utilized option in our clinical armamentarium, and further studies are needed to guide us in this area.


Quantifying Parkinson's disease severity using mobile wearable devices and machine learning: the ParkApp pilot study protocol.
The clinical assessment of Parkinson's disease (PD) symptoms can present reliability issues and, with visits typically spaced apart 6 months, can hardly capture their frequent variability. Smartphones and smartwatches along with signal processing and machine learning can facilitate frequent, remote, reliable and objective assessments of PD from patients' homes. To investigate the feasibility, compliance and user experience of passively and actively measuring symptoms from home environments using data from sensors embedded in smartphones and a wrist-wearable device. In an ongoing clinical feasibility study, participants with a confirmed PD diagnosis are being recruited. Participants perform activity tests, including Timed Up and Go (TUG), tremor, finger tapping, drawing and vocalisation, once a week for 2 months using the Mobistudy smartphone app in their homes. Concurrently, participants wear the GENEActiv wrist device for 28 days to measure actigraphy continuously. In addition to using sensors, participants complete the Beck's Depression Inventory, Non-Motor Symptoms Questionnaire (NMSQuest) and Parkinson's Disease Questionnaire (PDQ-8) questionnaires at baseline, at 1 month and at the end of the study. Sleep disorders are assessed through the Parkinson's Disease Sleep Scale-2 questionnaire (weekly) and a custom sleep quality daily questionnaire. User experience questionnaires, Technology Acceptance Model and User Version of the Mobile Application Rating Scale, are delivered at 1 month. Clinical assessment (Movement Disorder Society-Unified Parkinson Disease Rating Scale (MDS-UPDRS)) is performed at enrollment and the 2-month follow-up visit. During visits, a TUG test is performed using the smartphone and the G-Walk motion sensor as reference device. Signal processing and machine learning techniques will be employed to analyse the data collected from Mobistudy app and the GENEActiv and correlate them with the MDS-UPDRS. Compliance and user aspects will be informing the long-term feasibility. The study received ethical approval by the Swedish Ethical Review Authority (Etikprövningsmyndigheten), with application number 2022-02885-01. Results will be reported in peer-reviewed journals and conferences. Results will be shared with the study participants.


An introduction to teamwork: findings from an evaluation of an interprofessional education experience for 1000 first-year health science students.
Effective interprofessional collaboration is an important factor in addressing health care needs and priorities. Educators and health care practitioners have argued that interprofessional education (IPE) is necessary to equip students with the knowledge, skills, attitudes, and behaviors to work collaboratively and ultimately deliver enhanced patient/client care. The University of Toronto has implemented an introductory IPE session for approximately 1000 health science students that focuses on teamwork. This session provides students with an opportunity to be sensitized to the client's perspective and become familiar with roles and perspectives of different professions. A mixed method pre/post research design was developed to examine changes in students' perceptions and attitudes regarding IPE following their participation in this session. This study also endeavored to explore the pedagogic effectiveness of this large-scale IPE session. Students completed pre and post surveys based on the Interprofessional Attitudes Questionnaire and the Interdisciplinary Education Perception Scale. A total of 399 surveys were matched for pre/post analysis, and 25 students participated in follow-up focus group interviews. Findings from this study reveal that a significant shift in many indicators occurred after this single intervention. Despite the large numbers of students, which meant a complex range of logistical factors to negotiate, our findings indicated that it is feasible to deliver a successful IPE session to a large cohort of first-year students. We suggest that the findings presented in this report can be of value to other interprofessional groups of course developers.


Mutational Testing in Gastrointestinal Stromal Tumor.
Targeted treatment has become a major modality in cancer management. Such cancer drugs are generally designed to treat tumors with certain genetic/genomic makeups. Mutational testing prior to prescribing targeted therapy is crucial in identifying who can receive clinical benefit from specific cancer drugs. Over the last two decades, gastrointestinal stromal tumors (GISTs) have evolved from histogenetically obscure to being identified as distinct gastrointestinal mesenchymal tumors with well-defined clinical and molecular characteristics, for which multiple lines of targeted therapies are available. Although the National Comprehensive Cancer Network (NCCN) strongly recommends mutational testing for optimal management of GIST, many GIST patients still have neither a mutation test performed or any mutation-guided cancer management. Here, we review the mutation-guided landscape of GIST, mutational testing methods, and the recent development of new therapies targeting GIST with specific mutations.


Chloride dependent amino acid transport in the human small intestine.
Carriers of beta amino acids and imino acids in the small intestine of rabbits and guinea pigs are chloride dependent, and a cotransport of chloride, sodium, and 2-methyl-aminoisobutyric acid has been shown. This study examines the chloride dependence of amino acid transport in the human small intestine. The steady state tissue uptake of amino acids, given as the ratio between substrate concentration in intracellular and extracellular water after 35 minutes incubation at 37 degrees C, was determined in mucosal biopsy specimens from the duodenum of patients undergoing diagnostic upper endoscopy and compared using one way analysis of variance. Uptake of leucine and alpha-methyl-D-glucoside in the duodenum and the distal ileum did not differ. The accumulation of all substrates was sodium dependent. In the absence of mucosal chloride the uptake of taurine, glycine, and 2-methyl-aminoisobutyric acid was significantly reduced while that of leucine and alpha-methyl-D-glucoside was unaffected and the reduction of beta alanine uptake not statistically significant. Uptake of 2-methyl-aminoisobutyric acid and proline showed mutual inhibition. Leucine did not reduce uptake of the beta amino acids. In conclusion, chloride dependent transport processes for 2-methyl-amino-isobutyric acid, taurine, and glycine are present in the human small intestine.


Origin of the blueshift of water molecules at interfaces of hydrophilic cyclic compounds.
Water molecules at interfaces of materials exhibit enigmatic properties. A variety of spectroscopic studies have observed a high-frequency motion in these water molecules, represented by a blueshift, at both hydrophobic and hydrophilic interfaces. However, the molecular mechanism behind this blueshift has remained unclear. Using Raman spectroscopy and ab initio molecular dynamics simulations, we reveal the molecular mechanism of the blueshift of water molecules around six monosaccharide isomers. In the first hydration shell, we found weak hydrogen-bonded water molecules that cannot have a stable tetrahedral water network. In the water molecules, the vibrational state of the OH bond oriented toward the bulk solvent strongly contributes to the observed blueshift. Our work suggests that the blueshift in various solutions originates from the vibrational motions of these observed water molecules.


Skin and soft tissue infections in immunocompetent patients.
The increasing incidence of skin and soft tissue infections requires family physicians to be familiar with the management of these conditions. Evidence of systemic infection, such as fever, tachycardia, and hypotension, is an indication for inpatient management. Urgent surgical referral is imperative for those with life-threatening or rapidly advancing infections. In patients with uncomplicated abscesses measuring less than 5 cm in diameter, surgical drainage alone is the primary therapeutic intervention. Wound irrigation using tap water has similar outcomes as irrigation using sterile water. When antimicrobials are indicated, choice of agents depends on local resistance and susceptibility patterns. In settings where suspicion of methicillin-resistant Staphylococcus aureus (MRSA) is low, beta-lactam antibiotics are the first-line treatments for uncomplicated skin and soft tissue infections without focal coalescence or trauma. When empiric coverage for MRSA is indicated and the infection is uncomplicated, oral agents, such as tetracyclines, trimethoprim/sulfamethoxazole, and clindamycin, are preferred. Vancomycin is the first-line agent for MRSA in hospitalized patients, and newer agents, such as linezolid, daptomycin, and tigecycline, should be reserved for patients who do not respond to or cannot tolerate vancomycin therapy. There are insufficient data to support eradicating the carrier state in patients with MRSA or their contacts with nasal mupirocin or antibacterial body washes. Standard infection-control precautions, including proper and frequent handwashing, are a mainstay of MRSA prevention.


Resting-state connectivity in the prodromal phase of schizophrenia: insights from EEG microstates.
Resting-state EEG microstates are thought to reflect the momentary local states and interactions of distributed neural networks in the brain. Several changes in resting-state EEG microstates have been described in acutely ill patients with schizophrenia, but it is not known whether these represent trait or state abnormalities. The present study aimed to investigate this issue by assessing EEG microstate characteristics in high-risk individuals (HR) and clinically stable first-episode patients with schizophrenia (SZ) with low symptom levels, compared to each other and healthy controls (HC). Participants were 18 HR, 18 SZ, and 22 HC subjects. 64-channel resting-state EEG recordings were used for microstate analyses. Microstates were clustered into four classes (A-D) according to their topography. Temporal parameters and topographies of microstates were compared among groups. Microstate class A displayed higher coverage and occurrence in HR than SZ and HC, while microstate class B covered significantly more time in SZ compared to both HR and HC. Microstate class B displayed an aberrant spatial configuration in SZ, and to a lesser extent also in HR, compared to HC, with patients exhibiting significantly higher activity in the vicinity of the left posterior cingulate. Microstate abnormalities observed in HR were similar to those previously reported in acutely ill patients with schizophrenia. Moreover, there was evidence that HR and SZ might share specific disturbances in brain functional connectivity. These findings raise the possibility that certain abnormalities in resting-state EEG microstates might be associated with an increased risk for psychosis.


Keratocyte fragments and cells utilize competing pathways to move in opposite directions in an electric field.
Sensing of an electric field (EF) by cells-galvanotaxis-is important in wound healing [1], development [2], cell division, nerve growth, and angiogenesis [3]. Different cell types migrate in opposite directions in EFs [4], and the same cell can switch the directionality depending on conditions [5]. A tug-of-war mechanism between multiple signaling pathways [6] can direct Dictyostelium cells to either cathode or anode. Mechanics of motility is simplest in fish keratocytes, so we turned to keratocytes to investigate their migration in EFs. Keratocytes sense electric fields and migrate to the cathode [7, 8]. Keratocyte fragments [9, 10] are the simplest motile units. Cell fragments from leukocytes are able to respond to chemotactic signals [11], but whether cell fragments are galvanotactic was unknown. We found that keratocyte fragments are the smallest motile electric field-sensing unit: they migrate to the anode, in the opposite direction of whole cells. Myosin II was essential for the direction sensing of fragments but not for parental cells, while PI3 kinase was essential for the direction sensing of whole cells but not for fragments. Thus, two signal transduction pathways, one depending on PI3K, another on myosin, compete to orient motile cells in the electric field. Galvanotaxis is not due to EF force and does not depend on cell or fragment size. We propose a "compass" model according to which protrusive and contractile actomyosin networks self-polarize to the front and rear of the motile cell, respectively, and the electric signal orients both networks toward cathode with different strengths.