Regulation of R7 and R8 differentiation by the spalt genes.
Photoreceptor development begins in the larval eye imaginal disc, where eight distinct photoreceptor cells (R1-R8) are sequentially recruited into each of the developing ommatidial clusters. Final photoreceptor differentiation, including rhabdomere formation and rhodopsin expression, is completed during pupal life. During pupation, spalt was previously proposed to promote R7 and R8 terminal differentiation. Here we show that spalt is required for proper R7 differentiation during the third instar larval stage since the expression of several R7 larval markers (prospero, enhancer of split mdelta0.5, and runt) is lost in spalt mutant clones. In R8, spalt is not required for cell specification or differentiation in the larval disc but promotes terminal differentiation during pupation. We show that spalt is necessary for senseless expression in R8 and sufficient to induce ectopic senseless in R1-R6 during pupation. Moreover, misexpression of spalt or senseless is sufficient to induce ectopic rhodopsin 6 expression and partial suppression of rhodopsin 1. We demonstrate that spalt and senseless are part of a genetic network, which regulates rhodopsin 6 and rhodopsin 1. Taken together, our results suggest that while spalt is required for R7 differentiation during larval stages, spalt and senseless promote terminal R8 differentiation during pupal stages, including the regulation of rhodopsin expression.


Bone density, body composition and menstrual history of sedentary female former gymnasts, aged 20-32 years.
Few studies have examined the effects of retirement from sports involving regular, high impact and weight bearing activity on bone mass. This cross-sectional study compared total body and regional areal bone mineral density (aBMD, g/cm(2)) within female former gymnasts and women who had never participated in structured sport or exercise, and explored relations between aBMD of these former gymnasts and their duration of retirement. Eighteen sedentary female former gymnasts (GYM) and 18 sedentary controls (CON) were recruited. GYM displayed a broad range of duration of retirement (3-12 years) and a wide age range (20-32 years). GYM and CON were paired individually to match for age, body mass and stature. GYM had commenced training at least 3 years pre-menarche and had trained post-menarche for 2 or more years. They had trained continuously for 5-12 years and had retired between age 14 and 22 years. Measurements of aBMD and body composition were made using dual energy X-ray absorptiometry (DXA). Group mean values of physical and skeletal characteristics were compared using paired t-tests. Linear regression was used to explore possible relations of aBMD within GYM to duration of retirement. GYM displayed a higher aBMD than CON at all measurement sites, which ranged in magnitude from 6% for the total body ( P=0.004), to 11% for the total femur ( P=0.006). Elevations of aBMD within GYM equated to T-scores ranging from +0.8 (arms) to +1.7 (legs). There were no differences in body composition or age of menarche between groups, although 11 of 18 GYM reported a history of irregular menses. There was no significant decline of aBMD with increasing duration of retirement in GYM. The results suggested that an elevated bone mass in female former gymnasts was retained during early adulthood, in spite of a cessation of training for up to 12 years.


Effect of short peptides containing lysine and epsilon-aminocaproic acid on fibrinolytic activity of plasmin and topoisomerase II action on supercoiled DNA.
Effects of eight short peptides containing lysine and epsilon-aminocaproic acid (EACA) on prolongation of the clot lysis time, as well as hemolytic and antibacterial activities were investigated. Interaction with plasmids pBR322 and pUC19 with the use of ethidium bromide assay and determination of influence on the activity of topoisomerase I and II were also tested. Examined compounds inhibited fibrinolytic activity of plasmin and five of them were more active than EACA. Amides of dipeptides were most active antifibrinolytics (IC50 < 0.2 mM). According to the obtained data, the significant inhibition of fibrinolytic activity of plasmin was not associated with hemolytic effects. Examined compounds did not show antibacterial activity (MIC > 512 mg/L). DNA binding effects determined with the use of ethidium bromide were weak for all peptides and similar to those observed with EACA. Six compounds inhibited topoisomerase II action on supercoiled DNA.


Sacral plexus disorder caused by a wooden toothpick in the rectum.
A 67-year-old man presented with a 2-month history of pain in his right buttock and lower limb. MRI depicted right L5/S1 lateral recess stenosis requiring surgical treatment; however, preoperative CT showed an approximately 7 cm long, thin, rod-shaped structure in the rectum, which was ultimately determined to be an accidentally ingested toothpick. It was removed surgically 6 days after diagnosis, because right leg pain worsened rapidly. The pain disappeared thereafter, and the symptoms have not recurred since. The pain might have been localised to the right buttock and posterior thigh in the early stages because the fine tip of the toothpick was positioned to the right of the anterior ramus of the S2 spinal nerve. Although sacral plexus disorder caused by a rectal foreign body is extremely rare, physicians should be mindful to avoid misdiagnosis.


Youth and Young Adult Blunt Use Predicts Progression to Other Nicotine Product Use in the United States.
Little cigars or cigarillos (LCCs) are frequently modified to smoke cannabis ("blunts") by youth and young adults. This study investigated whether young blunt users who are otherwise nicotine-naïve are more likely to initiate other tobacco products compared to never blunt users. Data were from four waves of the Truth Longitudinal Cohort (TLC), a national probability-based sample of youth and young adults (aged 15-24 years) in the United States (Wave 1: January-April 2017; Wave 2: February-May 2018; Wave 3: February-May 2019; Wave 4: September-December 2019). The sample was restricted to nicotine naïve respondents at Wave 1 with possible ever use of blunts (N = 5,284). Logistic regression analyses tested whether ever blunt use at Wave 1 predicted initiation of nicotine products by Wave 4, controlling for established risk factors. Compared to never-blunt users, ever users of blunts at Wave 1 had significantly higher odds of ever using cigars (OR: 4.74; 95% CI: 1.80-12.47; p = 0.002), e-cigarettes (OR: 4.66; 95% CI: 2.42-8.95; p < 0.001), cigarettes (OR: 3.02; 95% CI: 1.17-7.84, p = 0.023), or hookah (OR: 3.47; 95% CI: 1.07-11.29, p = 0.039) by Wave 4. Cannabis (never blunt) use by Wave 1 predicted ever use of e-cigarettes (OR: 3.45, 95% CI: 2.38-5.02, p < 0.001), cigarettes (OR: 3.81; 95% CI: 2.26-6.43, p < 0.001), or hookah (OR: 2.13; 95% CI: 1.12-4.05, p = 0.021) by Wave 4. Blunts are a point of nicotine initiation that places users at increased risk of progression to cigars, while the same relationship was not found for cannabis alone.


Symptom dimensions in the course of childhood-onset schizophrenia.
The symptom dimensions of childhood-onset schizophrenia (COS) are described by focussing on the clinical features of 44 patients at onset of illness during the first episode and at follow-up investigation 42 years after onset. All subjects were re-diagnosed according to DSM IV. The symptomatology was evaluated with the Positive and Negative Symptom Scale (PANSS) at onset and at follow-up. Two principal component factor analyses with varimax-rotation were applied to the complete items set of the PANSS. The frequencies of positive, negative, and global symptoms were compared longitudinally in an ANOVA-repeated measures design. The factor analysis revealed 5 orthogonal symptom dimensions (factors) at onset of psychosis: Cognition, social withdrawal, antisocial behaviour, excitement, and reality distortion. At follow-up a five-factor solution was found, too, but different dimensions emerged: a positive, negative, excitement, cognitive, and anxiety/depression component which fits to the 5-factor model of White et al. (1997). The first psychotic episode of EOS is accompanied with more unspecific symptoms such as social withdrawal and antisocial behavior. In the later stages of (COS) the structure of symptom dimensions changes to that known from adult-onset schizophrenia (AOS). The results indicate that COS and AOS are comparable nosological entities and that more than 3 dimensions are required to describe the relevant clinical symptom structure. Positive and global symptoms decreased significantly during the course of illness. The frequencies of negative symptoms did not change which demonstrates their disabling impact.


Autospecific gammadelta thymocytes that escape negative selection find sanctuary in the intestine.
alphabeta or gammadelta thymocytes whose T-cell receptors (TCRs) recognize endogenously expressed antigens (Ag) are autospecific and, thus, potentially self-reactive. In the thymus, such T cells are eliminated during T-cell development through a process known as negative selection. As a model of negative selection of gammadelta T cells, we have used G8 gammadelta-T cell transgenic mice, which express a gammadelta TCR that recognizes the nonpolymorphic MHC class I TL(b) molecule. Here, we demonstrate that negative selection of autospecific gammadelta T cells is almost complete in the adult thymus but is markedly attenuated in the neonatal thymus. A consequence of this attenuated negative selection is that potentially self-reactive gammadelta thymocytes are allowed to escape negative selection, undergo extrathymic differentiation, and find sanctuary in the intestinal epithelium. Interestingly, the ability of these potentially self-reactive gammadelta T cells to find sanctuary requires both the intestinal epithelial environment and the extrathymic presence of the self-Ag. The implications of these findings on the development and persistence of autoreactive T cells in autoimmune disease are discussed.


Investigating the long-term legacy of drought and warming on the soil microbial community across five European shrubland ecosystems.
We investigated how the legacy of warming and summer drought affected microbial communities in five different replicated long-term (>10 years) field experiments across Europe (EU-FP7 INCREASE infrastructure). To focus explicitly on legacy effects (i.e., indirect rather than direct effects of the environmental factors), we measured microbial variables under the same moisture and temperature in a brief screening, and following a pre-incubation at stable conditions. Specifically, we investigated the size and composition of the soil microbial community (PLFA) alongside measurements of bacterial (leucine incorporation) and fungal (acetate in ergosterol incorporation) growth rates, previously shown to be highly responsive to changes in environmental factors, and microbial respiration. We found no legacy effects on the microbial community size, composition, growth rates, or basal respiration rates at the effect sizes used in our experimental setup (0.6 °C, about 30% precipitation reduction). Our findings support previous reports from single short-term ecosystem studies thereby providing a clear evidence base to allow long-term, broad-scale generalizations to be made. The implication of our study is that warming and summer drought will not result in legacy effects on the microbial community and their processes within the effect sizes here studied. While legacy effects on microbial processes during perturbation cycles, such as drying-rewetting, and on tolerance to drought and warming remain to be studied, our results suggest that any effects on overall ecosystem processes will be rather limited. Thus, the legacies of warming and drought should not be prioritized factors to consider when modeling contemporary rates of biogeochemical processes in soil.


An Australian approach to the policy translation of deliberated citizen perspectives on biobanking.
Deliberative public engagement is recommended for policy development in contested ethical areas. Scholars provide little guidance on how deliberative outputs can be translated to policy. This paper describes the processes we undertook to design a deliberative public forum for citizens to develop recommendations on biobanking that were adopted as health policy. The 4-day forum, held in 2008 in Perth, Western Australia, was designed in collaboration with academic experts. Deliberant recommendations were recorded in a formal report presented to policy-makers. Deliberations were audio-taped and transcribed. Translation involved transcript analyses, comparison of recommendations to other stakeholder views and post-forum consultations. Sixteen citizens made recommendations on ethical, legal and social issues related to biobanking. Most recommendations were translated into biobanking guidelines, with which Western Australia government health agencies must comply. The value of deliberative public participation in policy-making was most evident when trade-offs in competing interests, hopes and concerns were required. Translation issues included the impact of a small number of participants with limited socio-demographic diversity on procedural and policy legitimacy. Assessing the sufficiency of diversity in citizen representation was central to the deliberation-to-translation process. Institutional context facilitated the uptake of deliberation and translation processes. The use of these processes influenced policy substance and credibility among stakeholders and contributed to the state government directive that policy compliance be mandatory. We urge others to publish deliberation-to-translation processes so that best-practices may be identified.


Understanding Periodic and Non-periodic Chemistry in Periodic Tables.
The chemical elements are the "conserved principles" or "kernels" of chemistry that are retained when substances are altered. Comprehensive overviews of the chemistry of the elements and their compounds are needed in chemical science. To this end, a graphical display of the chemical properties of the elements, in the form of a Periodic Table, is the helpful tool. Such tables have been designed with the aim of either classifying real chemical substances or emphasizing formal and aesthetic concepts. Simplified, artistic, or economic tables are relevant to educational and cultural fields, while practicing chemists profit more from "chemical tables of chemical elements." Such tables should incorporate four aspects: (i) typical valence electron configurations of bonded atoms in chemical compounds (instead of the common but chemically atypical ground states of free atoms in physical vacuum); (ii) at least three basic chemical properties (valence number, size, and energy of the valence shells), their joint variation across the elements showing principal and secondary periodicity; (iii) elements in which the (sp)8, (d)10, and (f)14 valence shells become closed and inert under ambient chemical conditions, thereby determining the "fix-points" of chemical periodicity; (iv) peculiar elements at the top and at the bottom of the Periodic Table. While it is essential that Periodic Tables display important trends in element chemistry we need to keep our eyes open for unexpected chemical behavior in ambient, near ambient, or unusual conditions. The combination of experimental data and theoretical insight supports a more nuanced understanding of complex periodic trends and non-periodic phenomena.


Translation and validation of the Korean Confusion Assessment Method for the Intensive Care Unit.
Delirium is a common problem and associated with poor outcomes in intensive care unit (ICU) patients. Diagnosis of delirium in ICU patients is limited and usually underdiagnosed by physicians. The Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) is one of the most widely used screening methods for detection of ICU delirium. Our goal was to translate and validate the CAM-ICU for use in the Korean ICU setting. Translation of the CAM-ICU was done according to the guidelines suggested by the Translation and Cultural Adaptation Group. For validation and interrater reliability assessment of the Korean CAM-ICU, two nurses independently assessed delirium in ICU patients and the results were compared with the reference evaluation, which was done by a psychiatrist using the Diagnostic and Statistical Manual of Mental Disorders IV (DSM-IV). Twenty-two patients were evaluated by two nurses and one psychiatrist expert independently. During the study period, we have continuously educated study nurses. Based on DSM-IV criteria, 16 out of 22 (72.7%) patients developed delirium. The sensitivities of the two nurses' evaluations using the Korean CAM-ICU were 89.80% for nurse 1 and 77.40% for nurse 2. Their specificities were 72.40% and 75.80% and their overall accuracy was 83.33% and 88.37% respectively. The Korean CAM-ICU was done with reasonable interrater reliability between nurse 1 and nurse 2 (κ = 0.81, p < 0.001). The Korean CAM-ICU showed good validity and could be incorporated into clinical practice in Korean ICUs. ISRCTN: ISRCTN50265663.


Platelet adhesion and thrombus formation on subendothelium in platelets deficient in glycoproteins IIb-IIIa, Ib, and storage granules.
Patients whose platelets are deficient in glycoprotein (GP) Ib, IIb-IIIa (thrombasthenia), or granule substances (storage pool deficiency, SPD) were studied to define further the properties of platelets that mediate platelet adhesion and thrombus formation on subendothelium. Both nonanticoagulated and citrated blood were exposed to everted, de-endothelialized rabbit vessel segments under controlled flow conditions and shear rates varying from 650 to 3,300 sec-1. Morphometry was used to measure platelet thrombus dimensions and the percentage of the subendothelial surface covered with contact (C) or spread (S) platelets. Adhesion was defined as C + S. The results in SPD demonstrated (1) reduced thrombus dimensions in delta-SPD (pure dense granule deficiency) in proportion to the magnitude of the dense granule defect; (2) an even greater reduction in thrombus dimensions in patients with combined deficiencies of alpha and dense granules (alpha delta-SPD); and (3) impaired platelet adhesion at several conditions in alpha delta-SPD and, in delta-SPD, a hematocrit-dependent impairment of adhesion in citrated blood at 2,600 sec-1. In thrombasthenia, platelets were present as a monolayer on the subendothelial surface in both nonanticoagulated and citrated blood, indicating an absolute requirement for GPIIb-IIIa in promoting platelet-platelet interaction at all shear rates and perfusion times. Two types of abnormalities in platelet-vessel wall interactions were observed. In nonanticoagulated blood, the percentage of platelets in the C phase was consistently increased at all shear rates, but C + S values were normal. These observations indicate that platelets deficient in GPIIb-IIIa do not spread normally on the subendothelial surface exposed to nonanticoagulated blood. With citrated blood, the C + S value in thrombasthenia was reduced at both 800 and 2,600 sec-1, as in von Willebrand's disease, and a similar degree of reduction (about 50%) was observed in normal blood treated with a monoclonal antibody to GPIIb-IIIa. The findings, together with theoretical considerations, are consistent with an hypothesis that GPIIb-IIIa mediates the spreading of platelets on subendothelium following the initial attachment through GPIb and that GPIIb-IIIa may be considered an adhesion site on the platelet membrane. Abnormalities of GPIIb-IIIa may, depending on the conditions of study, result in either increased values of C platelets or decreased values of C + S. The results of the study further suggest that a complex interaction of platelet granule factors and membrane GP mediate platelet adhesion and thrombus formation.


The l(3)S12 locus of Drosophila melanogaster: heterochromatic position effects and stage-specific misexpression of the gene in P element transposons.
l(3)S12 is a vital locus whose function is required in embryos, early larvae, late pupae and oogenesis. We have identified a cold-sensitive allele, l(3)S12(3), and characterized conditional misexpression of the gene associated with this mutation as well as with several euchromatic insertions of l(3)S12+ transposons. Surviving cold-sensitive mutants as well as underexpression variants generated by P element transformation display a phenotypic syndrome that can include delayed development, abnormal bristle morphology, and female sterility. Using these phenotypes, defects in putative "early" and "late" l(3)S12 expression can be identified. The sensitivity of certain l(3)S12+ insertions to site-specific euchromatic position effect appears to be due to separation of the gene from an endogenous enhancer element during cloning. This enhancerless construct can be used to identify and perhaps to select "permissive" euchromatic sites, presumably adjacent to enhancer elements, which in some cases permit elevated production not only of the l(3)S12 message, but also of a P element-l(3)S12 fusion transcript. Certain of these permissive sites appear to control stage-specific expression, and we propose that this system may be used to identify, clone, and characterize such loci. Heterochromatic position effect on this locus has been demonstrated. Available evidence suggests that the l(3)S12 gene may be involved in protein synthesis, perhaps encoding a ribosomal protein.


Potential Mechanism of Action of 3'-Demethoxy-6-O-demethyl-isoguaiacin on Methicillin Resistant Staphylococcus aureus.
Bacterial infections represent one of the main threats to global public health. One of the major causative agents associated with high morbidity and mortality infections in hospitals worldwide is methicillin-resistant Staphylococcus aureus. Therefore, there is a need to develop new antibacterial agents to treat these infections, and natural products are a rich source of them. In previous studies, we reported that lignan 3'-demethoxy-6-O-demethylisoguaiacin, isolated and characterized from Larrea tridentate, showed the best activity towards methicillin-resistant S. aureus. Thus, the aim of this study was to determine the potential molecular mechanism of the antibacterial activity of 3'-demethoxy-6-O-demethylisoguaiacin against methicillin-resistant S. aureus using microarray technology. Results of microarray genome expression were validated by real-time polymerase chain reaction (RT-PCR). The genetic profile expression results showed that lignan 3'-demethoxy-6-O-demethylisoguaiacin had activity on cell membrane affecting proteins of the ATP-binding cassette (ABC) transport system causing bacteria death. This molecular mechanism is not present in any antibacterial commercial drug and could be a new target for the development of novel antibacterial agents.


Impacts of age and gender at the risk of underlying medical conditions and death in patients with avian influenza A (H7N9): a meta-analysis study.
The objective of our study was to conduct a series of analyses that examined the impacts of age and gender at the risk of underlying medical conditions (UMCs) and death in patients with influenza A (H7N9). We began by searching for potentially relevant articles in English or Chinese before February 28, 2018. Additionally, we reviewed our own files and reference lists of articles identified by this search. The association between death and UMCs was significant in H7N9 patients, with an OR of 1.49 (95% CI: 1.24-1.78). Subgroup analyses showed that having two or more UMCs of any type (OR: 2.24; P=0.044), chronic respiratory diseases (OR: 1.81; P=0.032), and chronic cardiovascular disease (OR: 1.63; P=0.013) had an association with increased fatality in H7N9 patients. Age (60 years or older) [adjusted OR (AOR): 1.86; P=0.032] and gender (male: AOR: 1.68, P=0.006; female: AOR: 1.88, P=0.044) were significantly associated with death in H7N9 patients with UMCs compared to H7N9 patients without any UMC. Stratification analyses found statistically significant increased death in H7N9 patients with UMCs who were 60 years of age and older (AOR: 2.72; P<0.001) and gender (male; AOR=1.64; P=0.033), compared to H7N9 patients without these respective conditions. Impacts of age are substantial and significant at the risk of UMCs and death in H7N9 patients. This analysis did not find a significant difference in gender comparisons. Efforts should particularly focus on reducing fatality rates in patients with combined risks from UMCs and other significant impact factor such as age (60 years or older).


Terpenoids and sterols from Saussurea cauloptera.
A total of 27 natural products were isolated from Saussurea cauloptera, including two new eudesmane-type sesquiterpenoids, the gerin derivatives 2 and 3, and one new ent-labdane diterpenoid, compound 9. The known compounds included six sesquiterpenoids, eleven triterpenoids, six sterols, and one lignan. Their structures were elucidated on the basis of extensive spectroscopic and mass-spectrometric analyses, as well as by X-ray crystallography in the case of gerin (1). The structurally related compounds 1-4 were found to exhibit strong inhibitory activities against human gastric carcinoma (SGC-7901) cells.


Translation and cross-cultural adaptation into Italian of the self-administered FLARE-RA questionnaire for rheumatoid arthritis.
The aim was to provide a translation into Italian with cross-cultural adaptation of the French FLARE-Rheumatoid Arthritis (RA) questionnaire, and to test its acceptability, feasibility, reliability and construct validity in a single-centre cohort study. The French version of the FLARE-RA questionnaire was cross-culturally adapted and translated into Italian following an established forward-backward translation procedure, with independent translations and backtranslations. To validate the Italian version we tested the internal validity with Cronbach's alpha, test-retest reliability with the intraclass correlation coefficient, agreement between assessments with Bland-Altman plots and construct validity with Spearman's correlation coefficients. The questionnaire was tested on 283 consecutive RA outpatients (mean age 56.1±13.9 years, 226/283 females, median disease duration 12.6 years ranging from 0.2 to 70.6). For the global score (11 items) the Cronbach's alpha coefficient was 0.94. The intraclass correlation coefficient was 0.87 (95% CI, 0.76-0.96). The correlation of FLARE-RA global score was 0.59 (95% CI, 0.50-0.66) with the Disease Activity Score on 28 joints, 0.63 (95% CI, 0.55-0.71) with the Simplified Disease Activity Index, 0.77 (95% CI, 0.71-0.83) with the RA Impact of Disease and 0.67 (95% CI, 0.59-0.73) with the Health Assessment Questionnaire. The Italian version of the FLARE-RA is feasible, brief and easy to administer. The translated and cross-cultural adapted showed accordingly to be valid and reliable. This questionnaire has some practical advantages, such as clarity, comprehensiveness, simplicity, and a minimum filling time. The development of cross-cultural adapted questionnaires in different languages is of pivotal importance to obtain standardized and comparable data across countries.


Single-Tooth Replacement Using Dental Implants Supporting All-Ceramic and Metal-Based Reconstructions: Results at 18 Months of Loading.
The aim of this study was to compare customized zirconia and titanium abutments with respect to survival rates and technical, biologic, and esthetic outcomes. A total of 28 patients with single implants were randomly assigned to 12 customized zirconia (test, AC) and 16 customized titanium (control, MC) abutments. Technical, biologic, and esthetic outcomes were assessed after a mean follow-up time of 18 months. No biologic complications were observed, and no statistical difference for the bone-to-implant distance was found at 18 months (AC -0.05 ± 0.51 mm vs MC -0.28 ± 0.77 mm; P = .40). A similar discoloration of the peri-implant mucosa was observed (ΔEAC 9.6 ± 5.4, ΔEMC 7.6 ± 5.3; P = .46). The mean papilla score values evaluation presented no statistically significant differences between the test and control groups (AC 2.07 ± 0.94, MC 1.96 ± 0.84). At 18 months, reconstructions based on zirconia and titanium abutments exhibited similar survival rates and similar clinical outcomes.


The S2-S3 Loop of Kv7.4 Channels Is Essential for Calmodulin Regulation of Channel Activation.
Kv7.4 (KCNQ4) voltage-gated potassium channels control excitability in the inner ear and the central auditory pathway. Mutations in Kv7.4 channels result in inherited progressive deafness in humans. Calmodulin (CaM) is crucial for regulating Kv7 channels, but how CaM affects Kv7 activity has remained unclear. Here, based on electrophysiological recordings, we report that the third EF hand (EF3) of CaM controls the calcium-dependent regulation of Kv7.4 activation and that the S2-S3 loop of Kv7.4 is essential for the regulation mediated by CaM. Overexpression of the mutant CaM1234, which loses the calcium binding ability of all four EF hands, facilitates Kv7.4 activation by accelerating activation kinetics and shifting the voltage dependence of activation leftwards. The single mutant CaM3, which loses the calcium binding ability of the EF3, phenocopies facilitating effects of CaM1234 on Kv7.4 activation. Kv7.4 channels co-expressed with wild-type (WT) CaM show inhibited activation when intracellular calcium levels increase, while Kv7.4 channels co-expressed with CaM1234 or CaM3 are insensitive to calcium. Mutations C156A, C157A, C158V, R159, and R161A, which are located within the Kv7.4 S2-S3 loop, dramatically facilitate activation of Kv7.4 channels co-expressed with WT CaM but have no effect on activation of Kv7.4 channels co-expressed with CaM3, indicating that these five mutations decrease the inhibitory effect of Ca2+/CaM. The double mutation C156A/R159A decreases Ca2+/CaM binding and completely abolishes CaM-mediated calcium-dependent regulation of Kv7.4 activation. Taken together, our results provide mechanistic insights into CaM regulation of Kv7.4 activation and highlight the crucial role of the Kv7.4 S2-S3 loop in CaM regulation.


Face validity of a synthetic papilla designed for biliary sphincterotomy training.
Background and study aims Endoscopic retrograde cholangiopancreatography (ERCP) is considered one of the most technically demanding endoscopic procedures. Still, limited data are available on simulators in ERCP training. Recently, the Boškoski-Costamagna ERCP Trainer was validated as a realistic training model by our study group. As an extension to this model, a novel synthetic papilla has been designed allowing to train biliary sphincterotomy. We aimed to determine the face validity of this synthetic papilla and its didactic value for training sphincterotomy. Methods Expert participants, each with more a than 2500 ERCPs lifetime experience, were invited to perform a biliary sphincterotomy and fill out a questionnaire on the realism of the procedure and the didactic value. Results A total of 40 ERCP experts were included, originating from 16 different countries. Experts' opinion on realism of performing a biliary sphincterotomy was rated with a median of 7 on a 10-point Likert scale, resemblance of the performed maneuvers 8 and tactile feedback 7. When asked if the cutting was perceived as realistic, experts rated a 6 and the cutting result was rated 8. The potential of the cutting papilla as a training tool for novices was rated 4 on a 4-point scale and there was a high agreement among the experts to include the papilla in the training of novices (rating 4). Conclusion This is the first synthetic papilla available for training sphincterotomy on the Boškoski-Costamagna ERCP Trainer and it demonstrates good face validity. ERCP experts highly agree on its didactic value and added value in the training curriculum of novice endoscopists.


Caregivers' Experience of Decision-Making regarding Diagnostic Assessment following Cognitive Screening of Older Adults.
Targeted screening for dementia among older adults in primary healthcare has potential benefits such as better clinical outcomes and the opportunity to access services. Cognitive screening can be followed up by further diagnostic assessment to determine a diagnosis of dementia. Unfortunately, the rates of accepting further diagnostic assessment following cognitive screening are low. The objective of this study was to explore the caregivers' decision-making process regarding uptake of diagnostic assessment following positive screening results. A qualitative design was employed, and interpretative phenomenological analysis was used to analyze the data. Three major themes in caregiver decision-making were identified: gathering information, protecting the patient, and balancing obligation and convenience in caregiving. These findings suggest that the decision-making process involved effort to process information through observations of the patient and that caregivers emphasized quality of life.


Strategic roadmap for an early diagnosis of Alzheimer's disease based on biomarkers.
The diagnosis of Alzheimer's disease can be improved by the use of biological measures. Biomarkers of functional impairment, neuronal loss, and protein deposition that can be assessed by neuroimaging (ie, MRI and PET) or CSF analysis are increasingly being used to diagnose Alzheimer's disease in research studies and specialist clinical settings. However, the validation of the clinical usefulness of these biomarkers is incomplete, and that is hampering reimbursement for these tests by health insurance providers, their widespread clinical implementation, and improvements in quality of health care. We have developed a strategic five-phase roadmap to foster the clinical validation of biomarkers in Alzheimer's disease, adapted from the approach for cancer biomarkers. Sufficient evidence of analytical validity (phase 1 of a structured framework adapted from oncology) is available for all biomarkers, but their clinical validity (phases 2 and 3) and clinical utility (phases 4 and 5) are incomplete. To complete these phases, research priorities include the standardisation of the readout of these assays and thresholds for normality, the evaluation of their performance in detecting early disease, the development of diagnostic algorithms comprising combinations of biomarkers, and the development of clinical guidelines for the use of biomarkers in qualified memory clinics.


Galectin-3 is required for resident microglia activation and proliferation in response to ischemic injury.
Growing evidence suggests that galectin-3 is involved in fine tuning of the inflammatory responses at the periphery, however, its role in injured brain is far less clear. Our previous work demonstrated upregulation and coexpression of galectin-3 and IGF-1 in a subset of activated/proliferating microglial cells after stroke. Here, we tested the hypothesis that galectin-3 plays a pivotal role in mediating injury-induced microglial activation and proliferation. By using a galectin-3 knock-out mouse (Gal-3KO), we demonstrated that targeted disruption of the galectin-3 gene significantly alters microglia activation and induces ∼4-fold decrease in microglia proliferation. Defective microglia activation/proliferation was further associated with significant increase in the size of ischemic lesion, ∼2-fold increase in the number of apoptotic neurons, and a marked deregulation of the IGF-1 levels. Next, our results revealed that contrary to WT cells, the Gal3-KO microglia failed to proliferate in response to IGF-1. Moreover, the IGF-1-mediated mitogenic microglia response was reduced by N-glycosylation inhibitor tunicamycine while coimmunoprecipitation experiments revealed galectin-3 binding to IGF-receptor 1 (R1), thus suggesting that interaction of galectin-3 with the N-linked glycans of receptors for growth factors is involved in IGF-R1 signaling. While the canonical IGF-1 signaling pathways were not affected, we observed an overexpression of IL-6 and SOCS3, suggesting an overactivation of JAK/STAT3, a shared signaling pathway for IGF-1/IL-6. Together, our findings suggest that galectin-3 is required for resident microglia activation and proliferation in response to ischemic injury.


MammaPrint 70-gene profile quantifies the likelihood of recurrence for early breast cancer.
Over the past few years, a variety of multigene expression profiles have been developed to improve prognostication for early stage breast cancer and reduce overtreatment with chemotherapy. MammaPrint is the only test cleared by the US Food and Drug Administration for the prognostication of early breast cancer. The MammaPrint assay examines the expression of 70 genes in the primary tumor to stratify patients diagnosed with early stage breast cancer into good and poor prognosis groups. This evaluation reviews the development of the 70-gene profile, including validation studies involving patients with node-negative and 1 - 3 node-positive disease, the conversion of the 70-gene profile to a high-throughput diagnostic test, and the continuing prospective MINDACT clinical trial. The MammaPrint assay should help to determine which women with early breast cancer could be spared adjuvant chemotherapy.


Optimization and Modeling of Material Removal Rate in Wire-EDM of Silicon Particle Reinforced Al6061 Composite.
The mechanical, physical and interfacial properties of aluminum alloys are improved by reinforcing the silicon carbide particles (SiCp). Machinability of such alloys by traditional methods is challenging due to higher tool wear and surface roughness. The objective of research is to investigate the machinability of SiCp reinforced Al6061 composite by Wire-Electrical Discharge Machining (wire-EDM). The effect of wire-EDM parameters namely current (I), pulse-on time (Ton), wire-speed (Ws), voltage (Iv) and pulse-off time (Toff) on material removal rate (MRR) is investigated and their settings are optimized for achieving the high MRR. The experiments are designed by using Taguchi L16 orthogonal arrays. The MRR obtained at different experiments are analyzed using statistical tools. It is observed that all the chosen process parameters showed significant influence of on the MRR with contribution of 27.39%, 22.08%, 21.32%, 15.76% and 12.94% by I, Iv, Toff, Ton and Ws, respectively. At optimum settings, the Wire-EDM resulted in MRR of 65.21 mg/min and 62.41 mg/min for samples with 4% and 8% SiCp. The results also indicated reinforcing SiCp upto 8% showed marginally low influence on MRR. Microstructural investigation of the cut surface revealed the presence of craters with wave pattern on its surface. The top surface of the crater is featured by the recast layers connecting adjacent craters. Further, the statistical model is developed using linear regression to predict the MRR (?2-73.65%) and its predicting accuracy is verified by the confirmation trials. The statistical model is useful for predicting the MRR for different settings of the process parameters. The optimized settings can be used to improve the machining productivity by increasing the MRR while machining of Al6061-SiCp (upto 8 wt. %) alloy by wire-EDM industries.


[Alcohol use by future physicians--medical and social problem].
The aim of the study was to assess the kind of alcohol disorders and their recurrence among students of medicine. Authors analysed 143 people using Alcohol Use Disorder Identification Test (AUDIT). It was found that 54.2% of students drink alcohol once a month or rarely, and 41.5% at least twice a month. The standard amount of alcohol for 53.1% of interviewed students were 1-2 standard drinks (10 g of ethanol), while every fourth student drinks 5 and more standard drinks. Hazardous drinking was found in 46.5% of studied subjects, three times more among men than women. The positive results of AUDIT obtained 11.9% of interviewed students. The percentage of people with alcohol problems found after AUDIT was 4.3%. Alcohol use among students of medicine is a medical and social problem according to performed studies.


Responding to the challenge. A status report on shared decision-making programs.
Interactive video programs have been developed to provide patients with tailored information about treatment choices for their medical conditions. The article provides a status report on the use and production of these programs. Evaluation results are presented in terms of patient satisfaction, patient preference shifts and impacts on clinical practice. The future of shared medical decision-making is discussed.


Variation in osmotic response to sustained mannitol administration.
Osmotic agents such as mannitol remain a mainstay in the management of cerebral edema and raised intracranial pressure. Some patients do not respond to sustained mannitol administration with the expected rise in serum osmolality, and this may correlate with lack of therapeutic efficacy. To examine the variation in osmotic response to mannitol therapy and identify factors associated with a lack of an osmotic response to sustained mannitol administration. Data on consecutive patients admitted to a Neurology/Neurosurgery Intensive Care Unit who received scheduled doses of mannitol for at least 48 h were extracted from a prospectively collected database. All patients received intravenous isotonic saline solutions and had serial measurements of serum sodium and osmolality, at least twice daily. Non-responders were defined using two thresholds, a rise in serum sodium of < or =1 or < or =5 mEq/l over the 48-hour period. The cohort included 167 patients the majority with intracerebral and subarachnoid hemorrhage and brain tumors. 73 patients (44%) did not respond to mannitol with a rise in sodium of > or =5 mEq/l, and 37 (22%) did not see a rise of 1 mEq/l over 48 h of treatment. There were minor differences between responders and non-responders (> or =5 mEq/l) in terms of age (56 +/- 15 vs. 48 +/- 14), total mannitol dose (0.9 +/- 0.2 vs. 0.7 +/- 0.2 g/kg), and cumulative fluid balance at 72 h (91 +/- 1653 vs. -610 +/- 1692 ml). Multivariate analysis found that younger age, lower weight-adjusted mannitol dose, and more negative fluid balance were associated with lack of osmotic response. A substantial proportion of patients receiving sustained mannitol do not manifest the expected osmotic response. This lack of response may correlate with the failure of clinical efficacy seen in a subgroup of patients, who then require alternate agents such as hypertonic saline. This association merits further exploration.


Prevalence of bacteriologically-confirmed pulmonary tuberculosis in urban Blantyre, Malawi 2019-20: Substantial decline compared to 2013-14 national survey.
Recent evidence shows rapidly changing tuberculosis (TB) epidemiology in Southern and Eastern Africa, with need for subdistrict prevalence estimates to guide targeted interventions. We conducted a pulmonary TB prevalence survey to estimate current TB burden in Blantyre city, Malawi. From May 2019 to March 2020, 115 households in middle/high-density residential Blantyre, were randomly-selected from each of 72 clusters. Consenting eligible participants (household residents ≥ 18 years) were interviewed, including for cough (any duration), and offered HIV testing and chest X-ray; participants with cough and/or abnormal X-ray provided two sputum samples for microscopy, Xpert MTB/Rif and mycobacterial culture. TB disease prevalence and risk factors for prevalent TB were calculated using complete-case analysis, multiple imputation, and inverse probability weighting. Of 20,899 eligible adults, 15,897 (76%) were interviewed, 13,490/15,897 (85%) had X-ray, and 1,120/1,394 (80%) sputum-eligible participants produced at least one specimen, giving 15,318 complete cases (5,895, 38% men). 29/15,318 had bacteriologically-confirmed TB (189 per 100,000 complete-case (cc) / 150 per 100,000 with inverse weighting (iw)). Men had higher burden (cc: 305 [95% CI:144-645] per 100,000) than women (cc: 117 [95% CI:65-211] per 100,000): cc adjusted odds ratio (aOR) 2.70 (1.26-5.78). Other significant risk factors for prevalent TB on complete-case analysis were working age (25-49 years) and previous TB treatment, but not HIV status. Multivariable analysis of imputed data was limited by small numbers, but previous TB and age group 25-49 years remained significantly associated with higher TB prevalence. Pulmonary TB prevalence for Blantyre was considerably lower than the 1,014 per 100,000 for urban Malawi in the 2013-14 national survey, at 150-189 per 100,000 adults, but some groups, notably men, remain disproportionately affected. TB case-finding is still needed for TB elimination in Blantyre, and similar urban centres, but should focus on reaching the highest risk groups, such as older men.


Promises and pitfalls in implementation science from the perspective of US-based researchers: learning from a pre-mortem.
Implementation science is at a sufficiently advanced stage that it is appropriate for the field to reflect on progress thus far in achieving its vision, with a goal of charting a path forward. In this debate, we offer such reflections and report on potential threats that might stymie progress, as well as opportunities to enhance the success and impact of the field, from the perspective of a group of US-based researchers. Ten mid-career extramurally funded US-based researchers completed a "pre-mortem" or a group brainstorming exercise that leverages prospective hindsight to imagine that an event has already occurred and to generate an explanation for it - to reduce the likelihood of a poor outcome. We came to consensus on six key themes related to threats and opportunities for the field: (1) insufficient impact, (2) too much emphasis on being a "legitimate science," (3) re-creation of the evidence-to-practice gap, (4) difficulty balancing accessibility and field coherence, (5) inability to align timelines and priorities with partners, and (6) overly complex implementation strategies and approaches. We submit this debate piece to generate further discussion with other implementation partners as our field continues to develop and evolve. We hope the key opportunities identified will enhance the future of implementation research in the USA and spark discussion across international groups. We will continue to learn with humility about how best to implement with the goal of achieving equitable population health impact at scale.


Escherichia coli limits Salmonella Typhimurium infections after diet shifts and fat-mediated microbiota perturbation in mice.
The microbiota confers colonization resistance, which blocks Salmonella gut colonization1. As diet affects microbiota composition, we studied whether food composition shifts enhance susceptibility to infection. Shifting mice to diets with reduced fibre or elevated fat content for 24 h boosted Salmonella Typhimurium or Escherichia coli gut colonization and plasmid transfer. Here, we studied the effect of dietary fat. Colonization resistance was restored within 48 h of return to maintenance diet. Salmonella gut colonization was also boosted by two oral doses of oleic acid or bile salts. These pathogen blooms required Salmonella's AcrAB/TolC-dependent bile resistance. Our data indicate that fat-elicited bile promoted Salmonella gut colonization. Both E. coli and Salmonella show much higher bile resistance than the microbiota. Correspondingly, competitive E. coli can be protective in the fat-challenged gut. Diet shifts and fat-elicited bile promote S. Typhimurium gut infections in mice lacking E. coli in their microbiota. This mouse model may be useful for studying pathogen-microbiota-host interactions, the protective effect of E. coli, to analyse the spread of resistance plasmids and assess the impact of food components on the infection process.


Metabolomics and cognition in African American adults in midlife: the atherosclerosis risk in communities study.
Clinical studies have shown alterations in metabolic profiles when patients with mild cognitive impairment and Alzheimer's disease dementia were compared to cognitively normal subjects. Associations between 204 serum metabolites measured at baseline (1987-1989) and cognitive change were investigated in 1035 middle-aged community-dwelling African American participants in the biracial Atherosclerosis Risk in Communities (ARIC) Study. Cognition was evaluated using the Delayed Word Recall Test (DWRT; verbal memory), the Digit Symbol Substitution Test (DSST; processing speed) and the Word Fluency Test (WFT; verbal fluency) at visits 2 (1990-1992) and 4 (1996-1998). In addition, Cox regression was used to analyze the metabolites as predictors of incident hospitalized dementia between baseline and 2011. There were 141 cases among 1534 participants over a median 17.1-year follow-up period. After adjustment for established risk factors, one standard deviation increase in N-acetyl-1-methylhistidine was significantly associated with greater 6-year change in DWRT scores (β=-0.66 words; P=3.65 × 10-4). Two metabolites (one unnamed and a long-chain omega-6 polyunsaturated fatty acid found in vegetable oils (docosapentaenoate (DPA, 22:5 n-6)) were significantly associated with less decline on the DSST (DPA: β=1.25 digit-symbol pairs, P=9.47 × 10-5). Two unnamed compounds and three sex steroid hormones were associated with an increased risk of dementia (all P<3.9 × 10-4). The association of 4-androstene-3beta, 17beta-diol disulfate 1 with dementia was replicated in European Americans. These results demonstrate that screening the metabolome in midlife can detect biologically plausible biomarkers that may improve risk stratification for cognitive impairment at older ages.


Tetraethylammonium modifies gap junctions between pancreatic beta-cells.
Gap junctions between pancreatic beta-cells were quantitatively assessed in freeze-fracture replicas of isolated rat islets of Langerhans incubated for 90 min with or without the potassium conductance blocker tetraethylammonium (TEA). The results show that TEA increases the median number of particles per beta-cell gap junction but not the frequency of gap junctions at both nonstimulating and threshold-stimulating concentrations of glucose. TEA increased the relative gap junctional area at both concentrations of glucose. TEA had no effect on insulin release at a basal concentration of glucose but potentiated that release at the threshold glucose level. Thus TEA modifies beta-cell gap junctions independently of its effect on insulin release. However, the junctional changes observed were greater when insulin release was also elevated.


Lactiplantibacillus brownii sp. nov., a novel psychrotolerant species isolated from sauerkraut.
A Gram-stain-positive, rod-shaped, facultatively anaerobic and homofermentative strain, named WILCCON 0030T, was isolated from sauerkraut (fermented cabbage) collected from a local market in the Moscow region of Russia. Comparative analyses based on 16S rRNA gene sequence similarity and whole genome relatedness indicated that strain WILCCON 0030T was most closely related to the type strains Lactiplantibacillus nangangensis NCIMB 15186T, Lactiplantibacillus daoliensis LMG 31171T and Lactiplantibacillus pingfangensis LMG 31176T. However, the average nucleotide identity and digital DNA-DNA hybridization prediction values with these closest relatives only ranged from 84.6 to 84.9 % and from 24.1 to 24.7 %, respectively, and were below the 95.0 and 70.0% thresholds for species delineation. Substantiated by further physiological and biochemical analyses, strain WILCCON 0030T represents a novel species within the genus Lactiplantibacillus for which we propose the name Lactiplantibacillus brownii sp. nov. (type strain WILCCON 0030T=DSM 116485T=LMG 33211T).


A nicking enzyme-assisted allosteric strategy for self-resetting DNA switching circuits.
The self-regulation of biochemical reaction networks is crucial for maintaining balance, stability, and adaptability within biological systems. DNA switching circuits, serving as basic units, play essential roles in regulating pathways, facilitating signal transduction, and processing biochemical reaction networks. However, the non-reusability of DNA switching circuits hinders its application in current complex information processing. Herein, we proposed a nicking enzyme-assisted allosteric strategy for constructing self-resetting DNA switching circuits to realize complex information processing. This strategy utilizes the unique cleavage ability of the nicking enzyme to achieve the automatic restoration of states. Based on this strategy, we implemented a self-resetting DNA switch. By leveraging the reusability of the DNA switch, we constructed a DNA switching circuit with selective activation characteristics and further extended its functionality to include fan-out and fan-in processes by expanding the number of functional modules and connection modes. Furthermore, we demonstrated the complex information processing capabilities of these switching circuits by integrating recognition, translation, and decision functional modules, which could analyze and transmit multiple input signals and realize parallel logic operations. This strategy simplifies the design of switching circuits and promotes the future development of biosensing, molecular computing, and nanomachines.


Effects of an odor or taste stimulus applied to an artificial teat on the suckling behavior of newborn dairy calves.
In their first days of life, dairy calves in artificial rearing systems often have difficulty using an artificial teat for feeding. We examined the age at which calves are able to stand up voluntarily and suckle as well as their suckling behavior when presented with a plain dry teat versus a dry teat modified with a presumably attractive odor or taste substance. Single-housed newborn dairy calves (n = 51) were presented for ten consecutive days with a two-minute two-choice test, in which suckling time was recorded for 1) a plain (control) teat versus a glucose-coated teat (taste test) and 2) a plain teat versus a teat with a "Freshly Cut Grass" odor (odor test). On average, the calves were able to stand up voluntarily and suckle from the second or third day of age on. The "Freshly Cut Grass" odor had no significant effect on their suckling behavior. In contrast, the calves showed a significant preference for suckling the glucose-coated teat and displayed a significantly longer total suckling time in the taste test compared to the odor test. There were no significant differences between sexes regarding suckling behavior. The results of the present study show that glucose had a significant effect on the calves' teat preference and significantly increased total suckling time with a dry artificial teat. As such, glucose may increase suckling motivation in non-efficient drinkers or ill calves with low motivation to suckle.


Economic determinants and dietary consequences of food insecurity in the United States.
This paper reviews recent research on the economic determinants and dietary consequences of food insecurity and hunger in the United States. The new Current Population Study (CPS) food insecurity and hunger measure shows that hunger rates decline sharply with rising incomes. Despite this strong relationship, confirmed in other national datasets, a one-to-one correspondence between poverty-level incomes and hunger does not exist. In 1995, 13.1% of those in poverty experienced hunger and half of those experiencing hunger had incomes above the poverty level. Panel data indicate that those who are often food insufficient are much more likely than food-sufficient households to have experienced recent events that stress household budgets, such as losing a job, gaining a household member or losing food stamps. Cross-sectional work also demonstrates the importance of food stamps because benefit levels are inversely related to food insufficiency. Concern for the dietary consequences of domestic food insufficiency is well placed; recent research shows that the odds of consuming intakes <50% of the recommended dietary allowance (RDA) are higher for adult women and elderly individuals from food-insufficient households. Preschoolers from food-insufficient households do not consume significantly lower amounts than those from food-sufficient households, but mean intakes for the rest of members in those very same households are significantly lower for the food insufficient. This research highlights the importance of food insecurity and hunger indicators, further validates the use of self-reported measures and points to areas of need for future research and interventions.


Approaching Genetics Through the MHC Lens: Tools and Methods for HLA Research.
The current SARS-CoV-2 pandemic era launched an immediate and broad response of the research community with studies both about the virus and host genetics. Research in genetics investigated HLA association with COVID-19 based on in silico, population, and individual data. However, they were conducted with variable scale and success; convincing results were mostly obtained with broader whole-genome association studies. Here, we propose a technical review of HLA analysis, including basic HLA knowledge as well as available tools and advice. We notably describe recent algorithms to infer and call HLA genotypes from GWAS SNPs and NGS data, respectively, which opens the possibility to investigate HLA from large datasets without a specific initial focus on this region. We thus hope this overview will empower geneticists who were unfamiliar with HLA to run MHC-focused analyses following the footsteps of the Covid-19|HLA & Immunogenetics Consortium.


Effect of mobile telephone reminders on treatment outcome in HIV: evidence from a randomised controlled trial in India.
To assess whether customised mobile phone reminders would improve adherence to therapy and thus decrease virological failure among HIV infected patients starting antiretroviral treatment (ART). Randomised controlled trial among HIV infected patients initiating antiretroviral treatment. Three diverse healthcare delivery settings in south India: two ambulatory clinics within the Indian national programme and one private HIV healthcare clinic. 631 HIV infected, ART naïve, adult patients eligible to initiate first line ART were randomly assigned to mobile phone intervention (n=315) or standard care (n=316) and followed for 96 weeks.. The intervention consisted of customised, interactive, automated voice reminders, and a pictorial message that were sent weekly to the patients' mobile phones for the duration of the study. The primary outcome was time to virological failure (viral load >400 copies/mL on two consecutive measurements). Secondary outcomes included ART adherence measured by pill count, death rate, and attrition rate. Suboptimal adherence was defined as mean adherence <95%. Using an intention-to-treat approach we found no observed difference in time to virological failure between the allocation groups: failures in the intervention and standard care arms were 49/315 (15.6%) and 49/316 (15.5%) respectively (unadjusted hazard ratio 0.98, 95% confidence interval 0.67 to 1.47, P=0.95). The rate of virological failure in the intervention and standard care groups were 10.52 and 10.73 per 100 person years respectively. Comparison of suboptimal adherence was similar between both groups (unadjusted incidence rate ratio 1.24, 95% CI 0.93 to 1.65, P=0.14). Incidence proportion of patients with suboptimal adherence was 81/300 (27.0%) in the intervention arm and 65/299 (21.7%) in the standard care arm. The results of analyses adjusted for potential confounders were similar, indicating no significant difference between the allocation groups. Other secondary outcomes such as death and attrition rates, and subgroup analysis also showed comparable results across allocation groups. In this multicentre randomised controlled trial among ART naïve patients initiating first line ART within the Indian national programme, we found no significant effect of the mobile phone intervention on either time to virological failure or ART adherence at the end of two years of therapy.Trial registration Current Controlled Trials ISRCTN79261738.


Chromatic adaptation in the underwater environment.
The effects of human visual chromatic adaptation underwater are usually reported in subjective terms. The present study aimed to generate data that could be discussed within the internationally recognized CIE color metric. Adaptation was assessed in two underwater environments (blue and green backgrounds). Divers were required to color match stimuli using either a Burnham-type colorimeter or the Munsell Limit Color Cascade. Comparisons of the chromaticity coordinates of matches made in air and underwater with those defined by color and luminance meters revealed an increased sensitivity to wavelengths approximately complementary to those of the background. Laboratory simulations of the underwater environments broadly replicated the field results. The data allow comparisons with complex mathematical models of chromatic adaptation that have been derived from orthodox experiments. The data might be helpful in quantifying the limits of human visual performance underwater.


Comparison of candesartan, enalapril, and their combination in congestive heart failure: randomized evaluation of strategies for left ventricular dysfunction (RESOLVD) pilot study. The RESOLVD Pilot Study Investigators.
We investigated the effects of candesartan (an angiotensin II antagonist) alone, enalapril alone, and their combination on exercise tolerance, ventricular function, quality of life (QOL), neurohormone levels, and tolerability in congestive heart failure (CHF). Seven hundred sixty-eight patients in New York Heart Association functional class (NYHA-FC) II to IV with ejection fraction (EF) <0.40 and a 6-minute walk distance (6MWD) <500 m received either candesartan (4, 8, or 16 mg), candesartan (4 or 8 mg) plus 20 mg of enalapril, or 20 mg of enalapril for 43 weeks. There were no differences among groups with regard to 6MWD, NYHA-FC, or QOL. EF increased (P=NS) more with candesartan-plus-enalapril therapy (0.025+/-0.004) than with candesartan alone (0.015+/-0.004) or enalapril alone(0.015+/-0.005). End-diastolic (EDV) and end-systolic (ESV) volumes increased less with combination therapy (EDV 8+/-4 mL; ESV 1+/-4 mL; P<0.01) than with candesartan alone (EDV 27+/-4 mL; ESV 18+/-3 mL) or enalapril alone (EDV 23+/-7 mL; ESV 14+/-6 mL). Blood pressure decreased with combination therapy (6+/-1/4+/-1 mm Hg) compared with candesartan or enalapril alone (P<0.05). Aldosterone decreased (P<0.05) with combination therapy (23.2+/-5.3 pg/mL) at 17 but not 43 weeks compared with candesartan (0.7+/-7.8 pg/mL) or enalapril (-0.8+/-11. 3 pg/mL). Brain natriuretic peptide decreased with combination therapy (5.8+/-2.7 pmol/L; P<0.01) compared with candesartan (4. 4+/-3.8 pmol/L) and enalapril alone (4.0+/-5.0 pmol/L). Candesartan alone was as effective, safe, and tolerable as enalapril. The combination of candesartan and enalapril was more beneficial for preventing left ventricular remodeling than either candesartan or enalapril alone.


Salmonellosis in orally infected specific pathogen-free C57B1 mice.
Specific pathogen-free C57B1 mice are 100 to 1,000 times as sensitive as CD-1 mice to intravenous or oral challenge by Salmonella enteritidis or S. gallinarum. Resistance to infection by S. pullorum was unaffected. Growth of Listeria monocytogenes and Mycobacterium bovis (BCG) in intravenously infected C57B1 mice was similar to that seen in CD-1 mice. Quantitative counts of viable S. enteritidis in the walls of the stomach, small intestine, cecum, and large intestine and in the corresponding intestinal contents showed that most of the oral challenge inoculum was rapidly inactivated so that, by 24 hr, less than 1% was still viable. Overnight starvation and pretreatment with bicarbonate solution increased the relative survival of the challenge approximately 10-fold. Despite the rapid and extensive inactivation of the oral inoculum within the normal intestine, significant numbers of salmonellae reached the liver and spleen by 48 hr, and this systemic infection was subsequently responsible for the death of a high proportion of the challenged animals.


Gag-specific CD8+ T lymphocytes recognize infected cells before AIDS-virus integration and viral protein expression.
CD8(+) T cells are a key focus of vaccine development efforts for HIV. However, there is no clear consensus as to which of the nine HIV proteins should be used for vaccination. The early proteins Tat, Rev, and Nef may be better CD8(+) T cell targets than the late-expressed structural proteins Gag, Pol, and Env. In this study, we show that Gag-specific CD8(+) T cells recognize infected CD4(+) T lymphocytes as early as 2 h postinfection, before proviral DNA integration, viral protein synthesis, and Nef-mediated MHC class I down-regulation. Additionally, the number of Gag epitopes recognized by CD8(+) T cells was significantly associated with lower viremia (p = 0.0017) in SIV-infected rhesus macaques. These results suggest that HIV vaccines should focus CD8(+) T cell responses on Gag.


Epidemiology of bloodstream infections in burn-injured patients: a review of the national burn repository.
Bloodstream infections (BSIs) are a major cause of morbidity and mortality in thermally injured patients. However, these infections have not been well defined in this patient population. Therefore, the authors performed a retrospective case-control study to characterize the epidemiology, microbiology, and outcomes of burn-associated BSIs. A retrospective review of all patients in the National Burn Repository (NBR) between the years 1981 and 2007 was performed. All cases that had infection listed under complications were included in this study. For each case, two randomly selected patients from the same time period served as controls. Patient demographic data, extent of %TBSA, and type of infection were extracted. Primary end point was mortality. Secondary endpoints were hospital length of stay (LOS), intensive care unit LOS, total ventilator days, and hospital charges. Further analysis of the data involved case-matching patients by TBSA deciles, adjustment for the effects of TBSA and other potential confounders, and a sensitivity analysis of the effects of including or excluding sites that might have failed to consistently capture BSI information. A total of 11,793 patients (3931 cases and 7862 control) were included in the study. Of cultures revealing a Gram-positive organism, Staphylococcus aureus (32%) was the most common. From samples where isolation of a Gram-negative species occurred, Pseudomonas aeruginosa (35%) was more prominent. Infected patients were older (40.9 vs 32.8, P < .05) and had higher %TBSA (22.2 vs 7.9, P < .05). BSI was associated with significantly higher mortality (21.9% vs 3.09%), hospital LOS (47.4 vs 8.8 days) intensive care unit LOS (30.8 vs 2.6 days), ventilator days (29.2 vs 1.4 days), and hospital charge ($339,909.91 vs $33,272.43); P < .001 for all values. On evaluation of case-matched controls, mortality was higher for patients with BSI only <50% TBSA strata. Conclusions were unaffected by adjustment for TBSA and other possible confounders and was not influenced by possible failure of some sites to consistently capture BSI information. Development of BSI in hospitalized burn patients is associated with significant increases in morbidity, mortality, and resource utilization.


Comparison of European isolates of viruses causing hemorrhagic fever with renal syndrome by a neutralization test.
Different virus isolates causing hemorrhagic fever with renal syndrome (HFRS) were compared using a neutralization test. Patient convalescent sera and antisera prepared in rabbits were used to compare Puumala-related Hantavirus isolates from Finland, Sweden, Belgium, and the USSR. The majority of European isolates were indistinguishable from each other using both homologous rabbit antisera and patient convalescent sera. The European isolates of HFRS were also compared with prototype Hantaan (the etiologic agent of Korean hemorrhagic fever). The one-way cross-reaction between the Hantaan and Puumala viruses, previously described using human convalescent sera tested by indirect immunofluorescence and immunoprecipitation, was also seen by the neutralization test.


GeLL: a generalized likelihood library for phylogenetic models.
Phylogenetic models are an important tool in molecular evolution allowing us to study the pattern and rate of sequence change. The recent influx of new sequence data in the biosciences means that to address evolutionary questions, we need a means for rapid and easy model development and implementation. Here we present GeLL, a Java library that lets users use text to quickly and efficiently define novel forms of discrete data and create new substitution models that describe how those data change on a phylogeny. GeLL allows users to define general substitution models and data structures in a way that is not possible in other existing libraries, including mixture models and non-reversible models. Classes are provided for calculating likelihoods, optimizing model parameters and branch lengths, ancestral reconstruction and sequence simulation. http://phylo.bio.ku.edu/GeLL under a GPL v3 license.


[Clinical aspects of traumatic uveitis and causes of enucleation].
The authors analyze 37 cases with grave traumatic uveitis eventuating in enucleation. Terms of enucleation after the injury, type of the injury, clinical picture of chronic uveitis, echographic and electrophysiological findings are presented. In ten patients signs of sensitization to ocular tissues were detected in various periods after the injury. Sensitization was detected by the leukocyte migration inhibition test (LMIT). The authors regard the results as a response to release of tissue antigens in the bloodflow because of impairment of the blood-eye barrier caused by the trauma. The authors claim that the results of LMIT in traumatic uveitis may be regarded as an additional, together with the clinical symptoms, criterion among indications to enucleation.


Detection of divergent genes in microbial aCGH experiments.
Array-based comparative genome hybridization (aCGH) is a tool for rapid comparison of genomes from different bacterial strains. The purpose of such analysis is to detect highly divergent or absent genes in a sample strain compared to an index strain. Development of methods for analyzing aCGH data has primarily focused on copy number abberations in cancer research. In microbial aCGH analyses, genes are typically ranked by log-ratios, and classification into divergent or present is done by choosing a cutoff log-ratio, either manually or by statistics calculated from the log-ratio distribution. As experimental settings vary considerably, it is not possible to develop a classical discriminant or statistical learning approach. We introduce a more efficient method for analyzing microbial aCGH data using a finite mixture model and a data rotation scheme. Using the average posterior probabilities from the model fitted to log-ratios before and after rotation, we get a score for each gene, and demonstrate its advantages for ranking and detecting divergent genes with enlarged specificity and sensitivity. The procedure is tested and compared to other approaches on simulated data sets, as well as on four experimental validation data sets for aCGH analysis on fully sequenced strains of Staphylococcus aureus and Streptococcus pneumoniae. When tested on simulated data as well as on four different experimental validation data sets from experiments with only fully sequenced strains, our procedure out-competes the standard procedures of using a simple log-ratio cutoff for classification into present and divergent genes.


NasT-mediated antitermination plays an essential role in the regulation of the assimilatory nitrate reductase operon in Azotobacter vinelandii.
Azotobacter vinelandii is a well-studied model system for nitrogen fixation in bacteria. Regulation of nitrogen fixation in A. vinelandii is independent of NtrB/NtrC, a conserved nitrogen regulatory system in proteobacteria. Previous work showed that an ntrC mutation in A. vinelandii resulted in a loss of induction of assimilatory nitrate and nitrite reductases encoded by the nasAB operon. In addition to NtrC, several other proteins, including NasT, a protein containing a potential RNA-binding domain ANTAR (AmiR and NasR transcription antitermination regulators), have been implicated in nasAB regulation. In this work, we characterize the sequence upstream of nasA and identify several DNA sequence elements, including two potential NtrC binding sites and a putative intrinsic transcriptional terminator upstream of nasA that are potentially involved in nasAB regulation. Our analyses confirm that the nasAB promoter, P(nasA), is under NtrC control. However, unlike NtrC-regulated promoters in enteric bacteria, P(nasA) shows high activity in the presence of ammonium; in addition, the P(nasA) activity is altered in the nifA gene mutation background. We discuss the implication of these results on NtrC-mediated regulation in A. vinelandii. Our study provides direct evidence that induction of nasAB is regulated by NasT-mediated antitermination, which occurs within the leader region of the operon. The results also support the hypothesis that NasT binds the promoter proximal hairpin of nasAB for its regulatory function, which contributes to the understanding of the regulatory mechanism of ANTAR-containing antiterminators.


Morphological characteristics and abundance of prokaryotes associated with gills in mangrove brachyuran crabs living along a tidal gradient.
Due to the chemico-physical differences between air and water, the transition from aquatic life to the land poses several challenges for animal evolution, necessitating morphological, physiological and behavioural adaptations. Microbial symbiosis is known to have played an important role in eukaryote evolution, favouring host adaptation under changing environmental conditions. We selected mangrove brachyuran crabs as a model group to investigate the prokaryotes associated with the gill of crabs dwelling at different tidal levels (subtidal, intertidal and supratidal). In these animals, the gill undergoes a high selective pressure, finely regulating multiple physiological functions during both animal submersion under and emersion from the periodical tidal events. We hypothesize that similarly to other marine animals, the gills of tidal crabs are consistently colonized by prokaryotes that may quantitatively change along the environmental gradient driven by the tides. Using electron microscopy techniques, we found a thick layer of prokaryotes over the gill surfaces of all of 12 crab species from the mangrove forests of Saudi Arabia, Kenya and South Africa. We consistently observed two distinct morphotypes (rod- and spherical-shaped), positioned horizontally and/or perpendicularly to the gill surface. The presence of replicating cells indicated that the prokaryote layer is actively growing on the gill surface. Quantitative analysis of scanning electron microscopy images and the quantification of the bacterial 16S rRNA gene by qPCR revealed a higher specific abundance of prokaryote cells per gill surface area in the subtidal species than those living in the supratidal zone. Our results revealed a correlation between prokaryote colonization of the gill surfaces and the host lifestyle. This finding indicates a possible role of prokaryote partnership within the crab gills, with potential effects on animal adaptation to different levels of the intertidal gradient present in the mangrove ecosystem.


Epigenetics, hippocampal neurogenesis, and neuropsychiatric disorders: unraveling the genome to understand the mind.
In mature, differentiated neurons in the central nervous system (CNS), epigenetic mechanisms--including DNA methylation, histone modification, and regulatory noncoding RNAs--play critical roles in encoding experience and environmental stimuli into stable, behaviorally meaningful changes in gene expression. For example, epigenetic changes in mature hippocampal neurons have been implicated in learning and memory and in a variety of neuropsychiatric disorders, including depression. With all the recent (and warranted) attention given to epigenetic modifications in mature neurons, it is easy to forget that epigenetic mechanisms were initially described for their ability to promote differentiation and drive cell fate in embryonic and early postnatal development, including neurogenesis. Given the discovery of ongoing neurogenesis in the adult brain and the intriguing links among adult hippocampal neurogenesis, hippocampal function, and neuropsychiatric disorders, it is timely to complement the ongoing discussions on the role of epigenetics in mature neurons with a review on what is currently known about the role of epigenetics in adult hippocampal neurogenesis. The process of adult hippocampal neurogenesis is complex, with neural stem cells (NSCs) giving rise to fate-restricted progenitors and eventually mature dentate gyrus granule cells. Notably, neurogenesis occurs within an increasingly well-defined "neurogenic niche", where mature cellular elements like vasculature, astrocytes, and neurons release signals that can dynamically regulate neurogenesis. Here we review the evidence that key stages and aspects of adult neurogenesis are driven by epigenetic mechanisms. We discuss the intrinsic changes occurring within NSCs and their progeny that are critical for neurogenesis. We also discuss how extrinsic changes occurring in cellular components in the niche can result in altered neurogenesis. Finally we describe the potential relevance of epigenetics for understanding the relationship between hippocampal neurogenesis in neuropsychiatric disorders. We propose that a more thorough understanding of the molecular and genetic mechanisms that control the complex process of neurogenesis, including the proliferation and differentiation of NSCs, will lead to novel therapeutics for the treatment of neuropsychiatric disorders.


Effects of zopiclone on the sleep of chronic insomniacs.
The purpose of this study was to evaluate the short- and intermediate-term (21 days) effectiveness, as well as the carryover and withdrawal effects of 7.5 mg of zopiclone, a new short-acting hypnotic agent. 6 patients with chronic insomnia, ranging in age from 33 to 57, participated in the study. Both EEG sleep recordings and subjective rating scales were used to evaluate the drug. Zopiclone increased the total sleep time throughout its 3-week administration period although only during the short-term period was this increase statistically above baseline. There was no carryover effect nor rebound insomnia upon drug withdrawal. The drug significantly increased the duration of stage 2 sleep, decreased stages 3 and 4 sleep, and had no effect on the duration of REM sleep. There were no reported side effects.


Hypokinesia in Huntington's disease co-occurs with cognitive and global dysfunctioning.
Besides chorea, hypokinesia is an important motor disturbance in Huntington's disease (HD) but its clinical, neuropsychiatric, and cognitive functioning correlates are largely unknown. This cross-sectional study investigates correlates of hypokinesia in HD and its effect on global functioning. Among 150 HD gene carriers, 96 patients were clinically motor manifest. Hypokinesia was assessed using the motor section of the Unified Huntington's Disease Rating Scale and global functioning was measured using the Total Functioning Capacity (TFC) scale. Neuropsychiatric measures included the Apathy Scale and the Composite International Diagnostic Interview for diagnosis of depression. The Mini Mental State Examination (MMSE) and a composite executive cognitive measure were used to assess global and executive cognitive functioning, respectively. Compared with 45 patients with no or mild hypokinesia, 51 patients with moderate to severe hypokinesia showed a significant difference in most clinical and neuropsychiatric variables and had worse cognitive functioning scores. However, using forward logistic regression analysis, poor executive cognitive functioning was the only independent correlate of hypokinesia (OR 7.33; 95% CI: 2.82-19.0; P < 0.001). Hypokinesia score was inversely associated with the TFC score (P < 0.001), also after adjusting for chorea, use of antipsychotics, apathy, and global and executive cognitive functioning. In conclusion, the presence of moderate to severe hypokinesia in HD patients co-occurs with executive cognitive dysfunction and adversely affects global functioning.


Health-related quality of life and symptom burden of epithelioid hemangioendothelioma patients: a global patient-driven Facebook study in a very rare malignancy.
Purpose: Epithelioid hemangioendothelioma (EHE) is an ultra-rare vascular sarcoma with unique clinical features. EHE is characterized by an unpredictable, often protracted, clinical course and highly variable clinical presentation. Due to difficulty recruiting ultra-rare cancer patients, health-related quality of life (HRQoL) of EHE patients has not yet been studied. The aim of this study was to assess EHE symptom burden and its impact on HRQoL and psychological distress.Methods: The study was initiated after EHE patients' foundations approached our research group to study HRQoL. Patients were recruited from the international EHE Facebook group from May through October 2018. Data were collected using the online PROFILES registry. Latent class cluster analysis was performed to identify groups based on frequently reported symptoms. Differences in HRQoL (EORTC-QLQ-C30) and psychological distress (Hospital Anxiety and Depression Scale) between symptom-based clusters were examined.Results: Among 115 EHE patients from 20 countries, three clusters were identified, with low-, intermediate- and high-symptom burden, respectively. Highly symptomatic patients (33%) had clinically relevantly lower scores on HRQoL compared to the other two groups (p < 0.001). These patients suffered mostly from pain, insomnia and fatigue. Symptom burden significantly correlated with reduced daily functioning and high levels of psychological distress. Only for highly symptomatic patients, HRQoL and symptom levels were worse compared to healthy individuals.Conclusion: For the first time, we studied HRQoL in a large international cohort of ultra-rare cancer patients with distinct clinical characteristics, enabled by collaboration with patients and use of social media. We showed a considerable number of EHE patients were highly symptomatic, with a significant impact on HRQoL and psychological distress.


Maintenance of valvular integrity with Impella left heart support: Results from the multicenter PROTECT II randomized study.
The Impella 2.5 axial flow pump, which is positioned across the aortic valve, is widely employed for hemodynamic support. The present study compared structural and functional integrity of the left heart valves in patients undergoing Impella vs intra-aortic balloon pump in the randomized PROTECT II trial. Transthoracic echocardiograms were performed at baseline, 1 and 3 months in 445 patients in the PROTECT II trial. Serial studies were analyzed by an independent echocardiography core laboratory for aortic and mitral valve structure and function, and left ventricular ejection fraction (LVEF). During Impella support there was no appreciable change in the degree of baseline valvular regurgitation. There were no cases of structural derangement of the mitral or aortic valve after use of the Impella device. At 90-day follow-up, there was an average 22% relative increase in LVEF from baseline (27% ± 9 vs. 33% ± 11, P < 0.001). The present echocardiographic analysis of the PROTECT II study confirms prior observations regarding the safety of the Impella 2.5 device with respect to mitral and aortic valve function.


Maternal weight gain in women with gestational diabetes mellitus.
The aim of the study was to examine the perinatal outcomes in gestational diabetes in women with body mass index (BMI)-adjusted gestational weight gain (GWG) according to the Institute of Medicine (IOM) 2009 recommendations. The clinic's database was used to analyze 675 singleton GDM pregnancy outcomes. GWG for the entire pregnancy was compared to IOM recommendations and adjusted for prepregnancy BMI categories: underweight <18.5; normal 18.5-24.9; overweight 25-29.9; and obese >30. The study group was divided into three categories: below IOM limits, within IOM limits and above IOM limits. Only 37% of women achieved the proper weight gain (n=256). Almost 30% of women with GDM (n=196) had an excessive weight gain. GWG above limits was associated with a significantly higher neonatal measurements and a higher rate of large-for-gestational-age neonates. In underweight and normal-prepregnancy-weight women, no relation between GWG and birth-weight percentile was noted. For the overweight and obese women, a positive significant correlation between GWG until GDM diagnosis and birth-weight percentile was noted (P=0.002), which was not present when GWG until delivery was considered. Limited weight gain in overweight and obese women with gestational diabetes mellitus results in favourable pregnancy outcomes.


Casein kinase II alteration precedes tau accumulation in tangle formation.
Previous studies have shown altered casein kinase II (CK-II) in Alzheimer's disease (AD). For the present study, the authors analyzed CK-II immunoreactivity at various stages of tangle formation using quantitative laser confocal microscopy and immunoelectron microscopy. AD hippocampal pyramidal cells without neurofibrillary tangles (NFTs) displayed 15% more anti-tau immunoreactivity (P less than 0.01) and 43% more anti-CKII immunolabeling than controls (P less than 0.001). In AD, tangle-bearing hippocampal neurons with strong anti-tau immunoreactivity (threefold increase from controls) showed a significant 22% increase in anti-CKII immunolabeling (P less than 0.01), compared with those without NFTs. Neurons with early neurofibrillary changes showed diffuse anti-CKII immunostaining in their cytoplasm and cell processes. In tangle-bearing neurons, in which a higher level of tau immunoreactivity was detected, anti-CKII immunolabeling was distributed along a fibrillar meshwork in cell bodies and processes. Linear regression analysis of anti-CKII and anti-tau immunoreactivity in AD showed a positive correlation (r = 0.53, P less than 0.001). At the ultrastructural level, anti-CKII was immunolocalized to the paired helical filaments (PHF) of the tangle-bearing neurons, as well as to PHF in neuropil threads and some dystrophic neurites in plaques. These results suggest a possible role for CK-II in tangle formation.


The use of conversational laughter by an individual with dementia.
While laughter has been shown to play a significant role in any social interaction; its conversational usage by a person with dementia has rarely been investigated. This paper will investigate the functional aspects of laughter during conversation in an individual with dementia. Conversation analysis is used in order to investigate laughter as a social phenomenon and to be able to investigate laughter in an empirical and authentic manner. The conversational strategies employed through laughter will be detailed and implications will be discussed.


Educational Intervention to Improve Code Status Discussion Proficiency Among Obstetrics and Gynecology Residents.
Obstetrics and gynecology (OB/GYN) residents receive little formal training in conducting code status discussions (CSDs). We piloted an educational intervention to improve resident confidence and competence at conducting CSDs. The OB/GYN residents at a single institution participated in a 3-part educational program. First, participants reviewed a journal article and completed an online module. Second, they received a didactic lecture followed by a resident-to-resident mock CSD. Finally, participants had a videotaped CSD with a standardized patient (SP). Pre- and postintervention surveys and performance evaluations were analyzed. A subgroup analysis was performed on those with completed data sets. Participants included 24 residents in postgraduate years (PGY) 1 to 4: 85% were female with a mean age of 29 years; 83% completed the entrance survey; 63% completed the SP CSD; and 42% completed of all parts of the intervention. Residents initially felt most prepared to discuss treatment options (3.3/5 on a Likert scale) and less prepared to discuss hospice, end-of-life care, and code status (2.2/5, 2.2/5, and 2.3/5, respectively). Performance during the resident-to-resident CSD was variable with scores (% of skills achieved) ranging from 27% to 93% (mean 64%). Performance at the SP encounter was similar with scores ranging from 40% to 73% (mean 56%). After intervention, residents felt more prepared for CSDs (3.7/5) and end-of-life care (3.9/5). The subgroup analysis failed to show a significant change in skill performance from the first to the second CSD. Participants found the components of this intervention helpful and reported improved confidence at conducting CSDs.


An automated tool for obtaining QSAR-ready series of compounds using semantic web technologies.
We describe an application (Collector) for obtaining series of compounds annotated with bioactivity data, ready to be used for the development of quantitative structure-activity relationships (QSAR) models. The tool extracts data from the 'Open Pharmacological Space' (OPS) developed by the Open PHACTS project, using as input a valid name of the biological target. Collector uses the OPS ontologies for expanding the query using all known target synonyms and extracts compounds with bioactivity data against the target from multiple sources. The extracted data can be filtered to retain only drug-like compounds and the bioactivities can be automatically summarised to assign a single value per compound, yielding data ready to be used for QSAR modeling. The data obtained is locally stored facilitating the traceability and auditability of the process. Collector was used successfully for the development of models for toxicity endpoints within the eTOX project. The software is available at http://phi.upf.edu/collector. The source code is located at https://github.com/phi-grib/Collector and is free for use under the GPL3 license. The web version is hosted at http://collector.upf.edu/. email@example.com. Supplementary data are available at Bioinformatics online.


Long-segment nerve allograft regeneration in the sheep model: experimental study and review of the literature.
Experimental work in the field of nerve allotransplantation has dealt with the feasibility of nerve allografts reconstructing nerve gaps. In the majority of studies, the nerve gap studied has been short, and some degree of regeneration has been achieved, even in the untreated allograft. To better approximate clinical nerve-allograft reconstruction, a series of long-segment (8-cm) nerve allografts were performed in the ovine model. Twenty outbred ewes were randomized into two experimental groups with four experimental conditions. Animals received nerve allografts treated under one of the following conditions: fresh nerve autograft, fresh nerve allograft, cold-preserved nerve autograft, or cold-preserved nerve allograft. The nerve grafts were examined and compared at 6 and 10 months, using histological, morphometric, and electro-physiologic analyses. The results of the study demonstrated that, while excellent regeneration occurred across the nerve autograft, the long nerve allograft could not support axonal elongation. Similarly, cold nerve preservation did not enhance regeneration. The sheep animal model allows for investigation of the long nerve gap and may be beneficial in a better correlation of experimental nerve transplantation with clinical conditions.


Gene-TEQ--a standardized comparative assessment of effects in the comet assay using genotoxicity equivalents.
Existing methods for the comparison of genotoxic effects in the comet assay bear considerable disadvantages such as the problem to link information about concentration dependence and severity of effects. Moreover, given the lack of standardized protocols and the use of various standards, it may be extremely difficult to compare different studies. In order to provide a method for standardized comparative assessment of genotoxic effects, the concept of genotoxicity equivalents (Gene-TEQ) was developed. As potential reference compounds for genotoxic effects, three directly acting (N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), methyl-methanesulfonate, and N-methyl-N-nitrosourea) and three indirectly acting (cyclophosphamide, dimethylnitrosamine, and 4-nitroquinoline-oxide) genotoxic substances were compared with respect to their cytotoxic (neutral red) and genotoxic (comet assay) concentration-response profiles in the permanent fish cell line RTL-W1. For further comparison, two sediment extracts from the upper Danube River were investigated as environmental samples. Based on the results of cytotoxicity and genotoxicity testing, MNNG was selected as the reference compound. At several exposure levels and durations, genotoxic effects of both the other pure substances and the environmental samples were calculated as percentages of the maximum MNNG effect and related to the absolute MNNG effect (EC values). Thus, genotoxicity equivalent factors (Gene-TEQs) relative to MNNG could be calculated. Gene-TEQs can easily be applied to pure substances, mixtures and field samples to provide information about their toxicity relative to the reference compound. Furthermore, the Gene-TEQ concept allows a direct comparison of environmental samples from different laboratories.


Targeting Cytokines, Pathogen-Associated Molecular Patterns, and Damage-Associated Molecular Patterns in Sepsis via Blood Purification.
Sepsis is characterized by a dysregulated immune response to infections that causes life-threatening organ dysfunction and even death. When infections occur, bacterial cell wall components (endotoxin or lipopolysaccharide), known as pathogen-associated molecular patterns, bind to pattern recognition receptors, such as toll-like receptors, to initiate an inflammatory response for pathogen elimination. However, strong activation of the immune system leads to cellular dysfunction and ultimately organ failure. Damage-associated molecular patterns (DAMPs), which are released by injured host cells, are well-recognized triggers that result in the elevation of inflammatory cytokine levels. A cytokine storm is thus amplified and sustained in this vicious cycle. Interestingly, during sepsis, neutrophils transition from powerful antimicrobial protectors into dangerous mediators of tissue injury and organ dysfunction. Thus, the concept of blood purification has evolved to include inflammatory cells and mediators. In this review, we summarize recent advances in knowledge regarding the role of lipopolysaccharides, cytokines, DAMPs, and neutrophils in the pathogenesis of sepsis. Additionally, we discuss the potential of blood purification, especially the adsorption technology, for removing immune cells and molecular mediators, thereby serving as a therapeutic strategy against sepsis. Finally, we describe the concept of our immune-modulating blood purification system.


Application of adaptive constructive neural networks to image compression.
The objective of the paper is the application of an adaptive constructive one-hidden-layer feedforward neural networks (OHL-FNNs) to image compression. Comparisons with fixed structure neural networks are performed to demonstrate and illustrate the training and the generalization capabilities of the proposed adaptive constructive networks. The influence of quantization effects as well as comparison with the baseline JPEG scheme are also investigated. It has been demonstrated through several experiments that very promising results are obtained as compared to presently available techniques in the literature.


HRTEM and STEM-HAADF characterisation of Au-TiO2 and Au-Al2O3 catalysts for a better understanding of the parameters influencing their properties in CO oxidation.
Gold catalysts supported on titania (Au-TiO(2)) and alumina (Au-Al(2)O(3)) were prepared by deposition-precipitation with urea and then activated before characterisation and reaction in CO oxidation, either by calcination in air at 500 °C or reduction under H(2) at 300 °C. Gold nanoparticles with average size in the range 2-4 nm were obtained, with calcination leading to larger gold nanoparticles than reduction. For Au-TiO(2), high activity was observed in CO oxidation at room temperature, independent of the activation treatment. This high activity could not be correlated to the presence of sub-nanometer gold clusters as reported in the literature, since they could not be detected by atomic-resolution high-angle annular dark-field scanning-transmission electron microscopy (HAADF-STEM). In the case of Au-Al(2)O(3), the performance in CO oxidation was found to strongly depend on the water content in the reaction gas feed and on the activation conditions, with calcination resulting in a poorly active catalyst whereas reduction gave activity of the same order as Au-TiO(2). A comparative study of Au-TiO(2) and Au-Al(2)O(3) by electron microscopy did not reveal distinct differences in the shapes of the Au nanoparticles, which are mostly flattened through interaction with the substrate in both samples, with side profile shapes varying from rounded hemispherical to well faceted truncated cubo-octahedra. More faceting is found for the samples calcined at 500 °C than reduced at 300 °C. Various possible parameters affecting the catalytic properties of gold in CO oxidation are discussed in the context of the relevant literature.