[Epidemiology and prevention of HCV infection in dialysis centers].
From many epidemiological studies comes out that haemodialysis patients are at high risk of hepatitis C virus (HCV) infection. This review shows the results of several prevalence and incidence studies carried out in dialysis centres. The prevalence of anti-HCV antibodies in haemodialysis patients varies from 20% to 38%. From these surveys comes out that serodiagnostic tests, usually employed in laboratory diagnosis (ELISA and RIBA), may underestimate the true prevalence of HCV infection. The very important risk factors in the haemodialysis units are blood transfusion and the length of time on haemodialysis. Nevertheless, these are not sufficient to explain completely the high prevalence of subjects HCV positive among the dialysis patients. Contamination of environmental surfaces, caused by a non correct application of the asepsis and disinfection techniques, could be, according to several authors, the more important cause of this infection among the dialysis patients.


Canadian Registry of Implantable Electronic Device Outcomes: Longer-term follow-up of the Riata lead under advisory.
The Riata lead advisory was announced in 2011 and has posed unique management challenges because of cable externalization and insulation abrasion, in some cases leading to sudden loss of defibrillation. The purpose of this study was to provide further data on the rate of electrical failure in a population of patients with a Riata lead under advisory. Using the Canadian Registry of Electronic Device Outcomes, prospective follow-up data were collected on 1352 Riata leads under advisory in 17 implantable cardioverter-defibrillator (ICD) implantation centers in Canada. Data on electrical and structural failure were collected, as well as periprocedural complications related to lead revision (abandonment or extraction). Of the 1352 leads included in this 12-month follow-up cohort, 110 leads were revised. Mean follow-up time was 10.4 ± 1.5 years. The 12-year rate of electrical failure for the 8Fr lead was 9.45%, whereas the 10-year failure rate for the 7Fr lead was 7.25% and was not found to be accelerating. Of the leads revised because of advisory only (patient preference, advisory related, electrical or structural failure), the majority were abandoned (n = 54 [76.0%]), whereas 17 (24.0%) were extracted using power tools. The rate of major complications in each group was 5.6% and 5.9%, respectively. This report provides further data on the risk of electrical lead failure over a long lead dwell time. The risk of failure exceeds the risk of periprocedural major complications, indicating that the risk-to-benefit ratio is favorable to revise the lead in appropriate clinical scenarios.


Exposures in the alumina and primary aluminium industry: an historical review.
We reviewed specific chemical exposures and exposure assessment methods relating to published and unpublished epidemiological studies in the alumina and primary aluminium industry. Our focus was to review limitations in the current literature and make recommendations for future research. Although some of the exposures in the smelting of aluminium have been well characterised, particularly in potrooms, little has been published regarding the exposures in bauxite mining and alumina refining. Past epidemiological studies in the industry have concentrated on the smelting of aluminium, with many limitations in the methodology used in their exposure assessment. We found that in aluminium smelting, exposures to fluorides, coal tar pitch volatiles (CTPV) and sulfur dioxide (SO2) have tended to decrease in recent years, but insufficient information exists for the other known exposures. Although excess cancers have been found among workers in the smelting of aluminium, the exposure assessment methods in future studies need to be improved to better characterise possible causative agents. The small number of cohort studies has been a factor in the failure to identify clear exposure-response relationships for respiratory diseases. A dose-response relationship has been recently described for fluoride exposure and bronchial hyper-responsiveness, but whether fluorides are the causative agent, co-agent or simply markers for the causative agent(s) for potroom asthma, remains to be determined. Published epidemiological studies and quantitative exposure data for bauxite mining and alumina refining are virtually non-existent. Determination of possible exposure-response relationships for this part of the industry through improved exposure assessment methods should be the focus of future studies.


Oral contraceptives and risk of endometriosis: a systematic review and meta-analysis.
Combined oral contraceptives (OCs) inhibit ovulation, substantially reduce the volume of menstrual flow and may hypothetically interfere with implantation of refluxed endometrial cells. The aim of this review is to establish if OC use influences the risk of endometriosis. We performed a MEDLINE search to identify all studies published in the last four decades (January 1970 to January 2010) in the English language on the relationship between OC exposure and risk of endometriosis. Two authors abstracted data on standardized forms. We identified 608 potentially relevant studies and 18 studies (6 cross-sectional, 7 case-control and 5 cohort) were selected. Pooling of the results derived from all the included reports independently from study design, yielded a common relative risk of 0.63 [95% confidence interval (CI), 0.47-0.85] for current OC users, 1.21 (95% CI, 0.94-1.56) for past users and 1.19 (95% CI, 0.89-1.60) for ever users. Methodological drawbacks, such as uncertain temporal relationship between exposure and outcome in cross-sectional studies and suboptimal selection of controls in case-control studies, limit the quality of the available evidence. The risk of endometriosis appears reduced during OC use. However, it is not possible to exclude the possibility that the apparent protective effect of OC against endometriosis is the result of postponement of surgical evaluation due to temporary suppression of pain symptoms. Confounding by selection and indication biases may explain the trend towards an increase in risk of endometriosis observed after discontinuation, but further clarification is needed. To date, the hypothesis of recommending OCs for primary prevention of endometriosis does not seem sufficiently substantiated.


Significant differences in maternal child-feeding style between ethnic groups in the UK: the role of deprivation and parenting styles.
Nonresponsive maternal child-feeding interactions, such as restricting, pressurising and emotional feeding, can affect the ability of a child to self-regulate intake and increase the risk of becoming overweight. However, despite findings that South Asian and Black children living in the UK are more likely to be overweight, UK research has not considered how maternal child-feeding style might differ between ethnic groups. The present study aimed to explore variations in maternal child-feeding style between ethnic groups in the UK, taking into account associated factors such as deprivation and parenting style. Six hundred and fifty-nine UK mothers with a child who was aged 5-11 years old completed a questionnaire. Items included ethnicity and demographic data, as well as copies of the Child Feeding Questionnaire, Parental Feeding Styles Questionnaire and Parenting Styles and Dimensions Questionnaire. Significant differences in perceived responsibility (P = 0.002), restriction (P = 0.026), pressure to eat (P = 0.045), instrumental feeding (P = 0.000) and emotional feeding (P = 0.000) were found between the groups. Mothers from South Asian backgrounds reported higher levels of pressure to eat, emotional feeding and indulgent feeding styles, whereas mothers from Chinese backgrounds reported greater perceived responsibility and restriction. Mothers from Black and White British backgrounds were not significantly higher with respect to any behaviour. Maternal child-feeding style was also associated with deprivation and parenting style, although these did not fully explain the data. Understanding cultural factors behind maternal child-feeding style, particularly around pressurising and indulgent feeding behaviours, may play an important part in reducing levels of children who are overweight and obese in the UK.


Implications of Pathologic Complete Response Beyond Mediastinal Nodal Clearance With High-Dose Neoadjuvant Chemoradiation Therapy in Locally Advanced, Non-Small Cell Lung Cancer.
To determine, in a retrospective analysis of a large cohort of stage III non-small cell lung cancer patients treated with curative intent at our institution, whether having a pathologic complete response (pCR) influenced overall survival (OS) or freedom from recurrence (FFR) in patients who underwent definitive (≥60 Gy) neoadjuvant doses of chemoradiation (CRT). At our institution, 355 patients with locally advanced non-small cell lung cancer were treated with curative intent with definitive CRT (January 2000-December 2013), of whom 111 underwent mediastinal reassessment for possible surgical resection. Ultimately 88 patients received trimodality therapy. Chi-squared analysis was used to compare categorical variables. The Kaplan-Meier analysis was performed to estimate OS and FFR, with Cox regression used to determine the absolute hazards. Using high-dose neoadjuvant CRT, we observed a mediastinal nodal clearance (MNC) rate of 74% (82 of 111 patients) and pCR rate of 48% (37 of 77 patients). With a median follow-up of 34.2 months (range, 3-177 months), MNC resulted in improved OS and FFR on both univariate (OS: hazard ratio [HR] 0.455, 95% confidence interval [CI] 0.272-0.763, P = .004; FFR: HR 0.426, 95% CI 0.250-0.726, P = .002) and multivariate analysis (OS: HR 0.460, 95% CI 0.239-0.699, P = .001; FFR: HR 0.455, 95% CI 0.266-0.778, P = .004). However, pCR did not independently impact OS (P = .918) or FFR (P = .474). Mediastinal nodal clearance after CRT continues to be predictive of improved survival for patients undergoing trimodality therapy. However, a pCR at both the primary and mediastinum did not further improve survival outcomes. Future therapies should focus on improving MNC to encourage more frequent use of surgery and might justify use of preoperative CRT over chemotherapy alone.


Discretized and aggregated: modeling dive depth of harbor seals from ordered categorical data with temporal autocorrelation.
Ordered categorical data are pervasive in environmental and ecological data, and often arise from constraints that require discretizing a continuous variable into ordered categories. A great deal of data have been collected toward the study of marine mammal dive behavior using satellite depth recorders (SDRs), which often discretize a continuous variable such as depth. Additionally, data storage or transmission constraints may also necessitate the aggregation of data over time intervals of a specified length. The categorization and aggregation create a time series of ordered multicategory counts for each animal, which present challenges in terms of statistical modeling and practical interpretation. We describe an intuitive strategy for modeling such aggregated, ordered categorical data allowing for inference regarding the category probabilities and a measure of central tendency on the original scale of the data (e.g., meters), along with incorporation of temporal correlation and overdispersion. The strategy extends covariate-specific cutpoint models for ordinal data. We demonstrate the method in an analysis of SDR dive-depth data collected on harbor seals in Alaska. The primary goal of the analysis is to assess the relationship of covariates, such as time of day, with number of dives and maximum depth of dives. We also predict missing values and introduce novel graphical summaries of the data and results.


Single-Dilution COVID-19 Antibody Test with Qualitative and Quantitative Readouts.
The coronavirus disease 2019 (COVID-19) global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to place an immense burden on societies and health care systems. A key component of COVID-19 control efforts is serological testing to determine the community prevalence of SARS-CoV-2 exposure and quantify individual immune responses to prior SARS-CoV-2 infection or vaccination. Here, we describe a laboratory-developed antibody test that uses readily available research-grade reagents to detect SARS-CoV-2 exposure in patient blood samples with high sensitivity and specificity. We further show that this sensitive test affords the estimation of viral spike-specific IgG titers from a single sample measurement, thereby providing a simple and scalable method to measure the strength of an individual's immune response. The accuracy, adaptability, and cost-effectiveness of this test make it an excellent option for clinical deployment in the ongoing COVID-19 pandemic.IMPORTANCE Serological surveillance has become an important public health tool during the COVID-19 pandemic. Detection of protective antibodies and seroconversion after SARS-CoV-2 infection or vaccination can help guide patient care plans and public health policies. Serology tests can detect antibodies against past infections; consequently, they can help overcome the shortcomings of molecular tests, which can detect only active infections. This is important, especially when considering that many COVID-19 patients are asymptomatic. In this study, we describe an enzyme-linked immunosorbent assay (ELISA)-based qualitative and quantitative serology test developed to measure IgG and IgA antibodies against the SARS-CoV-2 spike glycoprotein. The test can be deployed using commonly available laboratory reagents and equipment and displays high specificity and sensitivity. Furthermore, we demonstrate that IgG titers in patient samples can be estimated from a single measurement, enabling the assay's use in high-throughput clinical environments.


Screening and validation of differentially expressed genes in adipose tissue of patients with obesity and type 2 diabetes mellitus.
White adipose tissue (WAT) plays a pivotal role in the onset of type 2 diabetes mellitus (T2DM) and obesity. Despite its significance the underlying pathogenesis and key genes associated with it remain elusive. In our study, we screened the differentially expressed genes (DEGs) in intra-abdominal WAT of T2DM patients with obesity, as well as those with simple obesity, aiming to lay a foundational theory for an in-depth investigation of T2DM pathogenesis and the identification of novel therapeutic targets. Gene expression datasets (GSE16415 and GSE71416) were retrieved from the Gene Expression Omnibus (GEO) database. We employed R for screening DEGs and conducted a functional enrichment analysis using the Metascape database. Combined Lasso regression and Boruta feature selection algorithms were used to identify key DEGs. Subsequently, these were cross-verified using the GSE29231 dataset. Samples and medical records were collected from clinical study participants. The mRNA and protein expressions of the key DEGs were verified using qRT-PCR and western blotting, respectively. We discerned a total of 130 DEGs, with 40 being upregulated and 90 downregulated. Functional and pathway enrichment analyses illuminated that these genes are instrumental in mediating metabolite and energy production, neutrophil-mediated immunity, and other associated biological processes. This includes their involvement in the tricarboxylic acid cycle, glycolysis/gluconeogenesis, peroxisome proliferator-activated receptors, and other signalling pathways. Two genes, CIDEA and FSCN1 emerged as key DEGs. The low expression of CIDEA and high expression of FSCN1 in the T2DM and obesity group were verified in clinical samples (P < 0.05). We established that CIDEA and FSCN1 manifest significant differential expression in T2DM patients who are obese. This suggests their potential as risk assessment markers and therapeutic targets for T2DM.


Sports concussion and associated post-traumatic headache.
Despite an incidence of approximately 3.8 million sports-related concussions per year, the pathophysiological basis of this injury remains poorly understood. Associated post-traumatic headache, both acute and chronic, can also provide a unique treatment challenge for medical personnel. The presence of new onset or persistent headache following injury often complicates return to play decisions. It is also now evident that recurrent head trauma may be associated with the development of some chronic neurodegenerative disorders. Although anecdotal reports and consensus guidelines are utilized in the management of sports concussion and associated post-traumatic headache, further evidence-based data are needed. Improved prevention and management of this injury will occur with ongoing educational and research efforts. As such advances are made, it is imperative the headache specialist have continued understanding of this evolving field.


Phylogenetic framework for the phylum Tenericutes based on genome sequence data: proposal for the creation of a new order Mycoplasmoidales ord. nov., containing two new families Mycoplasmoidaceae fam. nov. and Metamycoplasmataceae fam. nov. harbouring Eperythrozoon, Ureaplasma and five novel genera.
The genus Mycoplasma, including species earlier classified in the genera Eperythrozoon and Haemobartonella, contains ~ 120 species and constitutes an extensively polyphyletic assemblage of bacteria within the phylum Tenericutes. Due to their small genome sizes and lack of unique characteristics, the relationships among the mycoplasmas/Tenericutes are not reliably discerned. Using genome sequences for 140 Tenericutes, their evolutionary relationships were examined using multiple independent approaches. Phylogenomic trees were constructed for 63 conserved proteins, 45 ribosomal proteins, three main subunits of RNA polymerase and 16S rRNA gene sequences. In all of these trees, Tenericutes species reliably grouped into four main clades designated as the "Acholeplasma", "Spiroplasma", "Pneumoniae" and "Hominis" clusters. These clades are also distinguished based on a similarity matrix constructed based on 16S rRNA gene sequences. Mycoplasma species were dispersed across 3 of these 4 clades highlighting their extensive polyphyly. In parallel, our comparative genomic analyses have identified > 100 conserved signature indels (CSIs) and 14 conserved signature proteins (CSPs), which are uniquely shared by the members of four identified clades, strongly supporting their monophyly and identifying them in molecular terms. Mycoplasma mycoides, the type species of the genus Mycoplasma, and a small number of other Mycoplasma species, formed a strongly supported clade within the "Spiroplasma" cluster. Nine CSIs and 14 CSPs reliably distinguish this clade from all other Mycoplasmatales species. The remainder of the Mycoplasmatales species are part of the "Pneumoniae" and "Hominis" clusters, which group together in phylogenetic trees. Here we are proposing that the order Mycoplasmatales should be emended to encompass only the Mycoplasma species within the "Spiroplasma" cluster and that a new order, Mycoplasmoidales ord. nov., should be created to encompass the other Mycoplasma species. The "Pneumoniae" and the "Hominis" clusters are proposed as two new families, Mycoplasmoidaceae fam. nov., which includes the genera Eperythrozoon, Ureaplasma, and the newly proposed genera Malacoplasma and Mycoplasmoides, and Metamycoplasmataceae fam. nov. to contain the newly proposed genera Metamycoplasma, Mycoplasmopsis, and Mesomycoplasma. The results presented here allow reliable discernment, both in phylogenetic and molecular terms, of the members of the two proposed families as well as different described genera within these families including members of the genus Eperythrozoon, which is comprised of uncultivable organisms. The taxonomic reclassifications proposed here, which more accurately portray the genetic diversity among the Tenericutes/Mycoplasma species, provide a new framework for understanding the biological and clinical aspects of these important microbes.


[Xanthogranulomatous pyelonephritis: experience at the General Hospital of Albacete. Report of 16 cases].
A retrospective study of 16 Xanthogranulomatous Pyelonephritis (XGP) cases seen during a 12-years period (1983-1994) at the Hospital General de Albacete was performed. An analysis is made of the clinical features, laboratory data and imaging diagnostic techniques, comparing our findings with those of several series published in the literature. The outcome of this study is that clinical and biological picture of XGP is characteristic: Middle-aged woman admitted to a Internal Medicine Service because a constitutional syndrome, with important weight loss, general poor health, acceleration of erythrocyte sedimentation rate (ESR), anemia, hepatic disfunction and renal destruction imagings, frequently with renal stone and urinary tract infection history. This characteristics can be useful to suspect a XGP.


Components of a "dynein regulatory complex" are located at the junction between the radial spokes and the dynein arms in Chlamydomonas flagella.
Previous studies of flagellar mutants have identified six axonemal polypeptides as components of a "dynein regulatory complex" (DRC). The DRC is though to coordinate the activity of the multiple flagellar dyneins, but its location within the axoneme has been unknown (Huang et al., 1982; Piperno et al., 1992). We have used improved chromatographic procedures (Kagami and Kamiya, 1992) and computer averaging of EM images (Mastronarde et al., 1992) to analyze the relationship between the DRC and the dynein arms. Our results suggest that some of the DRC components are located at the base of the second radial spoke in close association with the inner dynein arms. (a) Averages of axoneme cross-sections indicate that inner arm structures are significantly reduced in three DRC mutants (pf3 < pf2 < sup-pf-3 < wt). (b) These defects are more pronounced in distal/medial regions of the axoneme than in proximal regions. (c) Analysis of flagellar extracts by fast protein liquid chromatography and SDS-PAGE indicates that a specific dynein I2 isoform is missing in pf3 and reduced in pf2 and sup-pf-3. Comparison with ida4 and pf3ida4 extracts reveals that this isoform differs from those missing in ida4. (d) When viewed in longitudinal section, all three DRC mutants lack a crescent-shaped density above the second radial spoke, and pf3 axonemes lack additional structures adjacent to the crescent. We propose that the crescent corresponds in part to the location of the DRC, and that this structure is also directly associated with a subset of the inner dynein arms. This position is appropriate for a complex that is thought to mediate signals between the radial spokes and the dynein arms.


Differential accumulation of gangliosides by the brains of MPTP-lesioned mice.
Although gangliosides have been reported to enhance recovery from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced lesions of the substantia nigra, evidence as to whether the administered gangliosides actually reach this or any other site of lesion in the central nervous system (CNS) at which they putatively enhance recovery is lacking. Therefore, studies were carried out to determine the amount of 3H-labeled ganglioside that was accumulated by the brains of MPTP-treated mice as well as by brains of control mice. No significant difference in the accumulation of 3H-bovine brain gangliosides or 3H-GD1a was seen between lesioned and control brains up to 240 min after injection of the labeled lipids. However, significantly more label was associated with the brains of MPTP-treated mice compared to controls 120 and 240 min after the injection of 3H-GM1. Analysis of the lipids extracted from the brain of a 3H-GM1-treated mouse revealed that the majority of label was still associated with 3H-GM1, 240 min after its administration. Autoradiography of tissue sections from the brains of MPTP-treated mice injected with 3H-GM1 showed that label was present in the ventricular spaces of the brain. This observation suggests that the administered gangliosides are present in the cerebrospinal fluid, which indicates that they have the potential to reach the lesioned CNS site at which they putatively enhance recovery.


Co-inhibition of BCL-W and BCL2 restores antiestrogen sensitivity through BECN1 and promotes an autophagy-associated necrosis.
BCL2 family members affect cell fate decisions in breast cancer but the role of BCL-W (BCL2L2) is unknown. We now show the integrated roles of the antiapoptotic BCL-W and BCL2 in affecting responsiveness to the antiestrogen ICI 182,780 (ICI; Fulvestrant Faslodex), using both molecular (siRNA; shRNA) and pharmacologic (YC137) approaches in three breast cancer variants; MCF-7/LCC1 (ICI sensitive), MCF-7/LCC9 (ICI resistant), and LY2 (ICI resistant). YC137 inhibits BCL-W and BCL2 and restores ICI sensitivity in resistant cells. Co-inhibition of BCL-W and BCL2 is both necessary and sufficient to restore sensitivity to ICI, and explains mechanistically the action of YC137. These data implicate functional cooperation and/or redundancy in signaling between BCL-W and BCL2, and suggest that broad BCL2 family member inhibitors will have greater therapeutic value than targeting only individual proteins. Whereas ICI sensitive MCF-7/LCC1 cells undergo increased apoptosis in response to ICI following BCL-W+/-BCL2 co-inhibition, the consequent resensitization of resistant MCF-7/LCC9 and LY2 cells reflects increases in autophagy (LC3 cleavage; p62/SQSTM1 expression) and necrosis but not apoptosis or cell cycle arrest. Thus, de novo sensitive cells and resensitized resistant cells die through different mechanisms. Following BCL-W+BCL2 co-inhibition, suppression of functional autophagy by 3-methyladenine or BECN1 shRNA reduces ICI-induced necrosis but restores the ability of resistant cells to die through apoptosis. These data demonstrate the plasticity of cell fate mechanisms in breast cancer cells in the context of antiestrogen responsiveness. Restoration of ICI sensitivity in resistant cells appears to occur through an increase in autophagy-associated necrosis. BCL-W, BCL2, and BECN1 integrate important functions in determining antiestrogen responsiveness, and the presence of functional autophagy may influence the balance between apoptosis and necrosis.


A Conceptual Framework for Rescheduling Elective Pediatric Gastroenterology Procedures Following COVID-19 Pandemic Lockdown.
Elective surgical and endoscopic procedures were suspended nationwide during the March 2020 COVID-19 pandemic to minimize exposure and healthcare resource utilization. This resulted in an unprecedented backlog of procedures in most clinical practices including pediatrics. Our group developed an internal process toward the rational development of an algorithm prioritizing elective procedures. This was based on patient disease severity defined by the presence of alert symptoms, symptom severity for dysphagia and abdominal pain, and diagnostic investigation findings. The underlying rationale is to prioritize patients in whom suspected disease course would be greatest impacted by endoscopy. We developed a nurse phone call-based process utilizing REDCap®, identifying relevant symptoms categorized by severity, and a validated functional impairment questionnaire for abdominal pain. We abstracted key laboratory and radiological findings also categorized by severity. The order of priority of procedures was established on the basis of a 4-tiered system factoring both presence and severity of symptoms or prior diagnostic testing results. We present the framework that we have adopted toward prioritizing procedures with the assumption that it offers an objective methodology and that can be efficiently and more broadly applied to other similar practice scenarios. Our tool may have wide-ranging implications both in the current COVID-19 pandemic and in other scenarios of limited resource allocation and deserves further investigation.


TFM-Explorer: mining cis-regulatory regions in genomes.
DNA-binding transcription factors (TFs) play a central role in transcription regulation, and computational approaches that help in elucidating complex mechanisms governing this basic biological process are of great use. In this perspective, we present the TFM-Explorer web server that is a toolbox to identify putative TF binding sites within a set of upstream regulatory sequences of genes sharing some regulatory mechanisms. TFM-Explorer finds local regions showing overrepresentation of binding sites. Accepted organisms are human, mouse, rat, chicken and drosophila. The server employs a number of features to help users to analyze their data: visualization of selected binding sites on genomic sequences, and selection of cis-regulatory modules. TFM-Explorer is available at http://bioinfo.lifl.fr/TFM.


Proteome-scale purification of human proteins from bacteria.
The completion of the human genome project and the development of high-throughput approaches herald a dramatic acceleration in the pace of biological research. One of the most compelling next steps will be learning the functional roles of all proteins. Achievement of this goal depends in part on the rapid expression and isolation of proteins at large scale. We exploited recombinational cloning to facilitate the development of methods for the high-throughput purification of human proteins. cDNAs were introduced into a master vector from which they could be rapidly transferred into a variety of protein expression vectors for further analysis. A test set of 32 sequence-verified human cDNAs of various sizes and activities was moved into four different expression vectors encoding different affinity-purification tags. By means of an automatable 2-hr protein purification procedure, all 128 proteins were purified and subsequently characterized for yield, purity, and steps at which losses occurred. Under denaturing conditions when the His6 tag was used, 84% of samples were purified. Under nondenaturing conditions, both the glutathione S-transferase and maltose-binding protein tags were successful in 81% of samples. The developed methods were applied to a larger set of 336 randomly selected cDNAs. Sixty percent of these proteins were successfully purified under denaturing conditions and 82% of these under nondenaturing conditions. A relational database, FLEXProt, was built to compare properties of proteins that were successfully purified and proteins that were not. We observed that some domains in the Pfam database were found almost exclusively in proteins that were successfully purified and thus may have predictive character.


Molecular simulations of carbohydrates and protein-carbohydrate interactions: motivation, issues and prospects.
The characterization of the 3D structure of oligosaccharides, their conjugates and analogs is particularly challenging for traditional experimental methods. Molecular simulation methods provide a basis for interpreting sparse experimental data and for independently predicting conformational and dynamic properties of glycans. Here, we summarize and analyze the issues associated with modeling carbohydrates, with a detailed discussion of four of the most recently developed carbohydrate force fields, reviewed in terms of applicability to natural glycans, carbohydrate-protein complexes and the emerging area of glycomimetic drugs. In addition, we discuss prospectives and new applications of carbohydrate modeling in drug discovery.


Glucocorticosteroids for infants with biliary atresia following Kasai portoenterostomy.
Biliary atresia is a life-threatening disease characterised by progressive destruction of both intra- and extra-hepatic biliary ducts. The mainstay of treatment is Kasai portoenterostomy, as soon as the disease has been confirmed. Glucocorticosteroids are steroid hormones which act on the glucocorticoid receptor and have a range of metabolic and immunomodulatory effects. Glucocorticosteroids are used to improve the postoperative outcomes in infants who have undergone Kasai portoenterostomy. To assess the beneficial and harmful effects of glucocorticosteroid administration versus placebo or no intervention following Kasai portoenterostomy in infants with biliary atresia. We searched the Cochrane Hepato-Biliary Group Controlled Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL) in the Cochrane Library, MEDLINE Ovid, Embase Ovid, Science Citation Index Expanded (Web of Science), and online trial registries (last search: 20 December 2017) for randomised controlled trials. We included randomised clinical trials which assessed glucocorticosteroids for infants who have undergone Kasai portoenterostomy. For harm, we also considered quasi-randomised studies, observational studies, and case-control studies that were identified amongst the search results. We used standard methodological procedures expected by Cochrane. We assessed the risk of bias for each trial according to prespecified domains. We analysed data using both random-effects and fixed-effect models. We performed the analyses using Review Manager 5.3 and Trial Sequental Analysis software. We considered a P value of 0.025 or less, two-tailed, as statistically significant. We planned to calculate risk ratios (RRs) for dichotomous outcomes, and the mean difference (MD) for continuous outcomes. For all association measures, we planned to use 95% confidence intervals (CIs) as well as Trial Sequential Analysis-adjusted CIs. We used Trial Sequential Analyisis to control the risks of random errors; however, we were often unable to implement this beyond calculating the required information size as there were few trials and data. We assessed the certainty of the evidence using GRADE. We found two randomised controlled trials fulfilling the inclusion criteria of our review. The trials provided data for meta-analysis. We judged the two trials as trials at low risk of bias. The two trials randomised a total of 213 infants to glucocorticosteroids versus placebo. In our Trial Sequential Analysis, the required information size (that is, the meta-analytic sample size) was not reached for any outcome. Trials were funded by charities, public organisations, and received support from private sector companies, none of which seemed to have an interest in the outcome of the respective trials. The effect of glucocorticosteroids after Kasai portoenterostomy on all-cause mortality is uncertain; the confidence interval is consistent with appreciable benefit and harm (RR 1.00; 95% CI 0.14 to 6.90; low-certainty evidence). The results showed little or no difference in adverse effects between the use of glucocorticosteroids or placebo after Kasai portoenterostomy, however this analysis was based on a single trial and we have low certainty in the result (RR 1.02; 95% CI 0.87 to 1.20;). Available data suggest that the proportions of infants who do not clear their jaundice at six months is similar between the two groups (RR 0.89; 95% CI 0.67 to 1.17; low-certainty evidence). All-cause mortality or liver transplantation did not differ at two years between the two groups (RR 1.00; 95% CI 0.72 to 1.39; low-certainty evidence). There were no data regarding health-related quality of life.Our searches also yielded 19 observational studies, some of them containing limited information on harmful effects of glucocorticosteroid treatment. We presented the extracted information narratively. We identified one further ongoing trial with no currently available results. The two meta-analysed randomised clinical trials present insufficient evidence to determine the effects of using glucocorticosteroids versus placebo after Kasai portoenterostomy in infants with biliary atresia on any of the primary or secondary review outcomes. There is insufficient evidence to support glucocorticosteroid use in the postoperative management of infants with biliary atresia for long-term outcomes of all-cause mortality or liver transplantation. It is also unclear if glucocorticosteroids are able to reduce the numbers of infants who did not clear their jaundice by six months. Further randomised, placebo-controlled trials are required to be able to determine if glucocorticosteroids may be of benefit in the postoperative management of infants with biliary atresia treated with Kasai portoenterostomy. Such trials need to be conducted as multicentre trials.


Epigenetic Effects of Psychoactive Drugs.
Currently, and globally, we are facing the worst epidemic of psychoactive drug abuse resulting in the loss of hundreds of thousands of lives annually. Besides alcohol and opioid use and misuse, there has been an increase in illicit abuse of psychostimulants. Epigenetics is a relatively novel area of research that studies heritable alterations in gene expression. Long-term administration of psychoactive drugs may lead to transcriptional changes in brain regions related to drug-seeking behaviors and rewards that can be passed down transgenerationally. Epigenetic biomarkers such as DNA methylation and histone modifications contribute to disease diagnoses. This review aims to look at the epigenetic modifications brought forth by psychoactive drug abuse.


Use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae).
Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (</= 50 bp in size) can be considered promising tool for trematode species identification. Using the novel approach of molecular morphometrics that is based on ITS2 secondary structure homologies, phylogenetic relationships of the various isolates of fasciolid species have been discussed.


Prenatal programming of nephron number and blood pressure.
A low nephron number has been advocated to explain the association between intrauterine growth restriction (IUGR) and hypertension in later life. IUGR not only leads to a low birth weight but is also hypothesized to reprogram nephrogenesis, which results in a low nephron endowment. Several methods are used to estimate the total glomerular number, but only stereological techniques result in accurate (unbiased) and precise (reproducible) data. Several studies, both in humans and animal models that have used these methods indeed revealed that IUGR leads to a low nephron number. According to the hyperfiltration hypothesis, this reduction in renal mass is supposed to lead to glomerular hyperfiltration and hypertension in remnant nephrons with subsequent glomerular injury with proteinuria, systemic hypertension and glomerulosclerosis. Even though IUGR is associated with both a low nephron endowment and an increased risk of hypertension, only circumstantial evidence is available to support the hyperfiltration hypothesis after prenatal programming. A prerequisite for establishment of this association in long-term, prospective follow-up studies is the ability to estimate glomerular numbers in living human beings, for which a further advancement in radiological techniques is necessary. Only then can the association between nephron endowment and blood pressure in humans be studied more conclusively.


A new role for proteins subunits of RNase P: stabilization of the telomerase holoenzyme.
RNase P, an RNA-protein complex, is essential for processing tRNAs. Three of the ten protein subunits of Saccharomyces cerevisiae RNase P (and a related complex, RNase MRP) co-purify with yeast telomerase, another RNA-protein complex. The three telomerase-associated proteins, Pop1, 6 and 7, bind to TLC1, the RNA subunit of telomerase. In a recent study (Garcia et al. Nat Commun), we used temperature sensitive alleles of the essential POP genes to determine their role in telomerase biogenesis. At permissive temperature, pop mutant cells grow normally, and the abundance of most proteins, including protein subunits of telomerase, is similar to wild type (WT). However, telomeres are short, and the amount of the mature telomerase holoenzyme is low. Unlike the RNA subunit of RNase MRP, TLC1 is more abundant in pop cells and properly folded, except at the Cs2a/TeSS domain where the Pop proteins bind. These defects correlate with defective movement of TLC1 from the cytoplasm, where it associates with telomerase proteins, back to the nucleus where it lengthens telomeres. Thus, Pop proteins are needed for the stable association of telomerase proteins with TLC1, and their reduction sequesters mature telomerase in the cytoplasm, away from its nuclear substrates.


Genome duplication events have led to a diversification in the CPT I gene family in fish.
The enzyme carnitine palmitoyltransferase (CPT) I is a major regulator of mitochondrial fatty acid oxidation in vertebrates. Numerous genome duplication events throughout evolution have given rise to three (in mammals) or multiple (in fish) genetically and functionally different isoforms of this enzyme. In particular, these isoforms represent a diversification of kinetic and regulatory properties stemming from mutations at the genomic and proteomic levels. Phylogenetic reconstructions reveal a comprehensive view of the CPT I family in vertebrates and genomic modifications leading to structural changes in proteins and functional differences between tissues and taxa. In a model fish species (rainbow trout), the presence of five CPT I isoforms suggests repeated duplication events in bony fishes and salmonids. Subsequently, an array of nucleotide and amino acid substitutions in the isoforms may contribute to a tissue-specific and a previously observed species-specific difference in the IC(50) for malonyl-CoA. Moreover, all five isoforms are expressed in trout at the mRNA level in skeletal muscle, heart, liver, kidney, and intestine. In general, transcript levels of the beta-isoforms were higher in muscle tissues, while levels of the alpha-isoforms were higher in other tissues. Rainbow trout also exhibit developmental plasticity in relative mRNA expression of CPT I isoforms from fry to juvenile to adult stage. Thus the evolution of CPT I has resulted in a very diverse family of isoforms. These differences represent a degree of specificity in the ability of species to regulate function at the protein and tissue levels, which, in turn, may allow for precise control of lipid oxidation in individual tissues during physiological perturbations.


[The myxoma syndrome: "cardiac myxoma, cutaneous pigmented lesions and peripheral and endocrine neoplasms". Apropos 2 cases].
We present two cases, mother and son, with "syndrome myxoma". Both had cardiac myxoma and cutaneous pigmented lesions. The son had a cutaneous myxoma and the mother had been diagnosed as having Cushing's syndrome caused by adrenal adenoma. The "syndrome myxoma" is a systemic disease which causes cardiac, cutaneous, and breast myxomas, adrenal disease, testicle and pituitary tumours. Whenever it is diagnosed all first relatives should be tested for the syndrome. The patient should be re-examined every six-twelve months, due to frequent recurrence of cardiac myxoma.


An asymmetric attraction model for the diversity and robustness of cell arrangement in nematodes.
During early embryogenesis in animals, cells are arranged into a species-specific pattern in a robust manner. Diverse cell arrangement patterns are observed, even among close relatives. In the present study, we evaluated the mechanisms by which the diversity and robustness of cell arrangements are achieved in developing embryos. We successfully reproduced various patterns of cell arrangements observed in various nematode species in Caenorhabditis elegans embryos by altering the eggshell shapes. The findings suggest that the observed diversity of cell arrangements can be explained by differences in the eggshell shape. Additionally, we found that the cell arrangement was robust against eggshell deformation. Computational modeling revealed that, in addition to repulsive forces, attractive forces are sufficient to achieve such robustness. The present model is also capable of simulating the effect of changing cell division orientation. Genetic perturbation experiments demonstrated that attractive forces derived from cell adhesion are necessary for the robustness. The proposed model accounts for both diversity and robustness of cell arrangements, and contributes to our understanding of how the diversity and robustness of cell arrangements are achieved in developing embryos.


Prothrombotic response to norepinephrine infusion, mimicking norepinephrine stress-reactivity effects, is partly mediated by α-adrenergic mechanisms.
Stress-induced prothrombotic changes are mediated by the sympathetic nervous system and critically involved in mental triggering of acute coronary syndromes, but the underlying psychobiology is not fully understood. We tested the hypothesis that a norepinephrine (NE) infusion to mimic effects of stress-induced NE release on blood coagulation elicits prothrombotic changes and examined to what extent these would be mediated by an alpha-adrenergic mechanism. In a single-blind placebo-controlled within-subjects design, 24 middle-aged, non-smoking, non-obese and normotensive men participated in three experimental trials with an interval between one and two weeks. Each trial applied two sequential infusions of 1 and 15 min duration with varying substances [i.e., saline as placebo, the non-specific α-blocker phentolamine (2.5 mg/min), and NE (5 μg/min)]: trial 1=saline + saline; trial 2=saline + NE, and trial 3=phentolamine + NE. Plasma levels of clotting factor VIII activity (FVIII:C), fibrinogen, and D-dimer were assessed from blood samples collected immediately before and 1 min and 20 min after infusion procedures. Compared to saline + saline, saline + NE induced increases over time in FVIII:C, fibrinogen, and D-dimer levels. With phentolamine + NE, fibrinogen levels remained increased compared to saline + saline, but changes in FVIII:C and D-dimer levels were no more different. Coagulation changes did not differ between saline + NE and phentolamine + NE. NE infusion activates blood coagulation. The resulting prothrombotic state could be one psychobiological mechanism underlying mental triggering of acute coronary syndromes. Blockade of α-adrenergic receptors partly attenuated NE effects on coagulation and could be implied to have preventive potential in susceptible individuals.


Are platelet volume indices of clinical use in COVID-19? A systematic review.
The incidence of thrombotic complications is high in COVID-19 patients with severe disease. As key regulators of thrombus formation, platelets likely play a crucial role as mediators of severe acute respiratory syndrome coronavirus 2 associated pathogenesis. Studies have reported that parameters reflecting platelet size, known as platelet volume indices (PVI), are raised in patients with thrombosis and can predict poor outcomes. This systematic review evaluates the potential for PVI to be used as a predictor of COVID-19 morbidity and mortality. English and Chinese databases were searched electronically to identify studies reporting data on mean platelet volume, platelet distribution width or platelet-large cell ratio in COVID-19 patients. Included articles underwent a quality rating and descriptive narrative analysis. Thirty-two studies were included in the systematic review. The results show a general trend for PVI to be raised in severe COVID-19 patients and non-survivors, with 14 studies reporting significant differences of baseline PVI between severe and mild disease. Nonetheless, longitudinal studies showed varying PVI trends over the course of the disease and evidence for PVI to be associated with disease progression was limited. The quality rating of 12 studies was poor, 16 were rated fair and four were good. Most studies were retrospective in design, used small study populations and did not consider confounding factors that influence platelet volume. Studies also contained technical flaws in PVI measurement, limiting the reliability of the results. The evidence on the clinical usefulness of PVI is greatly limited by the lack of prospective evaluation, together with technical problems in measuring PVI. Carefully designed prospective studies are warranted. https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=304305, identifier CRD42022304305.


Body Compatible Thermometer Based on Green Electrolytes.
With the use of coordinated complexes between aliphatic diols and calcium chloride (CaCl2) as green electrolytes, a body compatible, ecofriendly and low-cost thermometer is successfully developed. This particular conductive liquid possesses unique features of ultrafast response and high sensitivity against temperature change. The influences of CaCl2 concentration and the category of aliphatic diols on conductivity change reveal that the thermal sensing abilities of such green electrolytes are positively relevant to the viscosity change along with temperature change. Owing to the advantages of stability, reliability, and security, the thermometer can implement long-term and continuous temperature monitoring, which can fully meet the requirements of application of medical monitors, diagnostics, and therapies. Moreover, the inherent advantages of thermometers, including satisfactory biocompatibility and nontoxicity, afford great promise for applications in invasive and inflammatory devices.


Lithiophilic Ag Nanoparticle Layer on Cu Current Collector toward Stable Li Metal Anode.
Intractable hurdles of low Coulombic efficiency and dendritic Li formation during a repeated deposition/stripping process hinder the commercial use of Li metal anode for next-generation battery systems. Achieving uniform Li nucleation is one of the effective strategies to address these issues, and it is of practical importance to realize this on a commercial Cu current collector that is lithiophobic. Herein, we design a nanostructured Ag lithiophilic layer on a Cu foil via an electroless plating process for a Li metal current collector. The deposition of lithiophilic Ag particles that are homogeneously distributed on the Cu foil can reduce the nucleation overpotential, realizing uniform Li nucleation and subsequently flat Li plating. As a result, a stable cycle stability of up to 360 h (1 mA cm-2) and an average Columbic efficiency of 94.5% for 100 cycles (1 mA cm-2) are achieved. Furthermore, CuAg full cells with LiFePO4 as a cathode exhibit good cycle performances and low polarization voltage. This approach provides another facile way for a stable lithium metal anode.


A non-neutralizing antibody broadly protects against influenza virus infection by engaging effector cells.
Hemagglutinin (HA) is the immunodominant protein of the influenza virus. We previously showed that mice injected with a monoglycosylated influenza A HA (HAmg) produced cross-strain-reactive antibodies and were better protected than mice injected with a fully glycosylated HA (HAfg) during lethal dose challenge. We employed a single B-cell screening platform to isolate the cross-protective monoclonal antibody (mAb) 651 from mice immunized with the HAmg of A/Brisbane/59/2007 (H1N1) influenza virus (Bris/07). The mAb 651 recognized the head domain of a broad spectrum of HAs from groups 1 and 2 influenza A viruses and offered prophylactic and therapeutic efficacy against A/California/07/2009 (H1N1) (Cal/09) and Bris/07 infections in mice. The antibody did not possess neutralizing activity; however, antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis mediated by natural killer cells and alveolar macrophages were important in the protective efficacy of mAb 651. Together, this study highlighted the significance of effector functions for non-neutralizing antibodies to exhibit protection against influenza virus infection.


Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps.
Differential interference contrast (DIC) microscopy is a common mode of biological light microscopy used to achieve maximal resolution and contrast with label-free, weakly absorbing specimens such as cells. Maintaining the polarization state of the illuminating light is essential for the technique, and this requirement can conflict with optical trapping. We describe how to optimize DIC imaging using a light-emitting diode illumination source in a microscope while integrating a dual optical trap into the set up. Every time a polarized light beam reflects off or transmits through a dichroic mirror in the beam path, its polarization state will change if it is not polarized exactly parallel (p) or perpendicular (s) to the plane of incidence. We observe wavelength-dependent optical rotation and depolarization effects in our illumination light upon reflection from/transmission through dichroic mirrors in the beam path, resulting in significant degradation of image quality. We describe a method to compensate for these effects by introducing quarter-waveplates and a laser clean-up filter into the imaging pathway. We show that this approach achieves a full recovery of image quality.


Cancer causing chewing habits and related oral lesions - A comparative study among females of socio-culturally different areas of West Bengal.
Smokeless tobacco, as well as areca-nut both, causes cancerous and precancerous lesions of the oral cavity. The traditional value of India, as well as West Bengal do not allow females for smoking, but there is no such disapproval for using smokeless tobacco or areca nut. Geographically and socio-culturally, the Northern part of West Bengal differs from its Southern part. This study aimed to assess the prevalence of different chewing habits, habit products, and habit-related different oral lesions among females in two socio-culturally different areas of West Bengal. A total of 222 women from areas of North Bengal and 173 women from areas of South Bengal aged 15 years and above were selected. A face-to-face interview was conducted using a structured questionnaire. An oral cavity examination was done to rule out any oral mucosal alterations caused by cancer-causing habits. Chi-square test or Fischer's exact tests were used to compare unpaired proportions as appropriate. In areas of the northern part of Bengal, 42.34% of females were chewers, and in areas of the southern part of Bengal 18.50% of females were chewers. Younger female chewers were more from areas of the northern part of Bengal. Women, residing in different areas of the northern part of Bengal had more habit-related oral lesions, compared to the southern part of Bengal. Special attention should be given to increasing awareness regarding chewing habit-related health hazards among females, especially in areas of North Bengal.


Repetition probability does not affect fMRI repetition suppression for objects.
Previously several functional magnetic resonance imaging (fMRI) studies point toward the role of perceptual expectations in determining adaptation or repetition suppression (RS) in humans. These studies showed that the probability of repetitions of faces within a block influences the magnitude of adaptation in face-related areas of the human brain (Summerfield et al., 2008). However, a current macaque single-cell/local field potential (LFP) recording study using objects as stimuli found no evidence for the modulation of the neural response by the repetition probability in the inferior temporal cortex (Kaliukhovich and Vogels, 2010). Here we examined whether stimulus repetition probability affects fMRI repetition suppression for nonface object stimuli in the human brain. Subjects were exposed to either two identical [repetition trials (RTs)] or two different [alternation trials (ATs)] object stimuli. Both types of trials were presented blocks consisting of either 75% [repetition blocks (RBs)] or 25% [alternation blocks (ABs)] of RTs. We found strong RS, i.e., a lower signal for RTs compared to ATs, in the object sensitive lateral occipital cortex as well as in the face-sensitive occipital and fusiform face areas. More importantly, however, there was no significant difference in the magnitude of RS between RBs and ABs in each of the areas. This is in agreement with the previous monkey single-unit/LFP findings and suggests that RS in the case of nonface visual objects is not modulated by the repetition probability in humans. Our results imply that perceptual expectation effects vary for different visual stimulus categories.


A novel approach for the determination of paracetamol based on the reduction of N-acetyl-p-benzoquinoneimine formed on the electrochemically treated pencil graphite electrode.
A novel approach was proposed for the simple and rapid electrochemical determination of paracetamol (PC) in the presence of uric acid in body fluids. The voltammetric determination of PC is based on the electrochemical reduction of N-acetyl-p-benzoquinoneimine formed simultaneously on the electrochemically treated pencil graphite (ETPG) electrode during the measurement. ETPG electrodes were prepared by the potential cycling between -0.3V and +2.0V in 0.1M H(3)PO(4) solution. The electrochemical performance of the ETPG electrode was evaluated by adsorptive transfer stripping differential pulse voltammetry (ATSDPV). The resulting sensor showed good performance for the determination of PC in human blood serum samples with a linear range of 0.05-2.5 μM and a highly reproducible response (RSD of 3.1%). The calculated detection limit was 2.5 nM (S/N=3). The proposed method does not require any sample pretreatment, prevents the interference of uric acid and allows the determination of PC directly in blood serum samples.


A numerical study of failure mechanisms in the cemented resurfaced femur: effects of interface characteristics and bone remodelling.
Failure mechanisms of the resurfaced femoral head include femoral neck fracture in the short-term and stress shielding and implant loosening in the long-term. In this study, finite element simulations of the resurfaced femur considering a debonded implant-cement interface, variable stem-bone interface conditions, and bone remodelling were used to study load transfer within the resurfaced femur and to investigate its relationship with known failure mechanisms. Realistic three-dimensional finite element models of an intact and resurfaced femur were used. Various conditions at the interface between the stem of the prosthesis and the bone were considered. Loading conditions included normal walking and stair climbing. For all stem-bone contact conditions, the tensile stresses in the cement mantle varied between 1 MPa and 5.4 MPa, except near the distal rim of the resurfacing component where they reached 5.4-7MPa. In the case of full stem-bone contact, high von Mises stresses (114-121MPa) were generated in the implant at the stem-cup junction. These stresses were considerably reduced (maximum von Mises stress, 76 MPa) where a gap was present at the stem-bone interface. Resurfacing led to strain shielding of the bone of the femoral head (20-75 per cent strain reductions) and periprosthetic bone resorption (50-80 per cent bone density reductions) for all interface stem-bone contact conditions. In the lateral femoral head and the proximal femoral shaft around the trochantric region, bone density reductions varied between 10 per cent and 50 per cent. Bone apposition was observed in the inferior-medial part of the femoral head and proximal femoral neck region. For full stem-bone contact, more load was transferred through the stem to the surrounding bone, exacerbating strain shielding. Although femoral hip resurfacing conserves bone stock at the primary operation, strain shielding and periprosthetic bone resorption might lead to eventual loosening over time. Post-operatively, the resurfacing procedure generated elevated strains (0.50-0.75 per cent strain) in the proximal femoral neck-component junction irrespective of the variation in interface conditions, indicating an initial risk of femoral neck fracture. Subsequent to bone remodelling, this strain concentration was considerably reduced (0.35-0.50 per cent strain), lowering the risk of neck fracture. In order to reduce the potential risk of neck fracture, patients should avoid activities which might induce high loading of the hip during the early post-operative period to allow the bone around the proximal femoral neck to remodel and heal.


Cyclosporin A directly inhibits human B-cell proliferation by more than a single mechanism.
Cyclosporin A (CsA) is a potent immunosuppressive agent that inhibits T-cell proliferation and lymphokine production. There is less information on the direct effect of CsA on B-cells. We investigated the proliferative responses of human tonsillar B-lymphocytes to a "T dependent" mitogen, pokeweed mitogen (PWM), and to a "T independent" mitogen, Staphylococcus aureus (SA). Both responses were strongly inhibited by CsA. Nonspecific cytotoxicity was ruled out, and the inhibition was not reversed by adding IL1, IL2, or BCGF individually or in combination. Maximal inhibition of the PWM response occurred when CsA was added early in the culture period. Cyclosporin A added 18 hours after the start of culture was less effective, and adding CsA after 36 hours resulted in only minimal inhibition. However, with SA as mitogen, addition after 36 hours still affected substantial inhibition. These results, on the time of action and resistance to reversal by exogenous growth factors, suggest that CsA can directly inhibit human B-cells by a mechanism similar to its action on T-lymphocytes, blocking an early event critical to entry into cell cycle, but an additional mechanism of inhibition later in the cell cycle may also operate when the proliferative signal is provided by the T-independent mitogen SA.


Sphingosine-1-phosphate protects against bisphosphonate‑induced HUVEC cell death via regulation of c-Jun‑N‑terminal kinase signaling.
Bisphosphonates (BPs) remain the most widely used and effective antiresorptive agents in the treatment of postmenopausal osteoporosis. In particular, nitrogen-containing BPs (N-BPs) are more potent at inhibiting bone resorption in vivo than simple BPs, but they are associated with a number of side-effects including increased endothelial cell apoptosis in patients with multiple myeloma. Sphingosine-1-phosphate (S1P), a sphingolipid metabolite, plays important roles in the regulation of cell growth, differentiation and programmed cell death as a multifunctional bioactive lipid mediator. The aim of this study was to elucidate the protective effect and the possible mechanism of S1P against BP-induced cell damage using human umbilical vein endothelial cells (HUVECs). HUVECs were treated with S1P for 1 h and then with BP including alendronate, zoledronate and risedronate. S1P protects HUVECs against BP-induced cell death and the protective effect was increased by S1P in a dose-dependent manner. S1P blocked BP-induced caspase-3 activation, nuclear factor-κB activation, c-Jun-N-terminal kinase (JNK) phosphorylation and DNA fragmentation. The blocking of JNK phosphorylation inhibited BP-induced caspase activation and HUVEC cell death. The present study demonstrates that S1P inhibits BP-induced endothelial cell death via regulation of JNK phosphorylation, and also suggests that S1P has the potential to be a therapeutic drug in various vascular diseases induced by BP.


Need for early diagnosis of mental and mobility changes in Wernicke encephalopathy.
Korsakoff syndrome is a chronic form of amnesia resulting from thiamine deficiency. The syndrome can develop from unrecognized or undertreated Wernicke encephalopathy. The intra-individual course of Wernicke-Korsakoff syndrome has not been studied extensively, nor has the temporal progression of gait disturbances and other symptoms of Wernicke encephalopathy. Here we present the detailed history of a patient whose acute symptoms of Wernicke encephalopathy were far from stable. We follow his mobility changes and the shifts in his mental status from global confusion and impaired consciousness to more selective cognitive deficits. His Wernicke encephalopathy was missed and left untreated, being labeled as "probable" Korsakoff syndrome. Patients with a history of self-neglect and alcohol abuse, at risk of or suffering with Wernicke encephalopathy, should receive immediate and adequate vitamin replacement. Self-neglecting alcoholics who are bedridden may have severe illness and probably active Wernicke encephalopathy. In these patients, mobility changes, delirium, or impaired consciousness can be an expression of Wernicke encephalopathy, and should be treated to prevent further damage from the neurologic complications of thiamine deficiency.


Combined exposure to X-irradiation followed by N-ethyl-N-nitrosourea treatment alters the frequency and spectrum of Ikaros point mutations in murine T-cell lymphoma.
Ionizing radiation is a well-known carcinogen, but its potency may be influenced by other environmental carcinogens, which is of practical importance in the assessment of risk. Data are scarce, however, on the combined effect of radiation with other environmental carcinogens and the underlying mechanisms involved. We studied the mode and mechanism of the carcinogenic effect of radiation in combination with N-ethyl-N-nitrosourea (ENU) using doses approximately equal to the corresponding thresholds. B6C3F1 mice exposed to fractionated X-irradiation (Kaplan's method) followed by ENU developed T-cell lymphomas in a dose-dependent manner. Radiation doses above an apparent threshold acted synergistically with ENU to promote lymphoma development, whereas radiation doses below that threshold antagonized lymphoma development. Ikaros, which regulates the commitment and differentiation of lymphoid lineage cells, is a critical tumor suppressor gene frequently altered in both human and mouse lymphomas and shows distinct mutation spectra between X-ray- and ENU-induced lymphomas. In the synergistically induced lymphomas, we observed a low frequency of LOH and an inordinate increase of Ikaros base substitutions characteristic of ENU-induced point mutations, G:C to A:T at non-CpG, A:T to G:C, G:C to T:A and A:T to T:A. This suggests that radiation doses above an apparent threshold activate the ENU mutagenic pathway. This is the first report on the carcinogenic mechanism elicited by combined exposure to carcinogens below and above threshold doses based on the mutation spectrum of the causative gene. These findings constitute a basis for assessing human cancer risk following exposure to multiple carcinogens.


Hepatitis C virus (HCV) core serotypes in chronic HCV infection.
Recently, two distinct hepatitis C virus (HCV) serologic types have been identified on the basis of amino acid variations in the core region. The two serologic types can readily discriminate between genotypes I-II-V (serotype 1) and III-IV (serotype 2), according to the Okamoto classification. We compared HCV core serotyping with genotyping with sera from 363 anti-HCV-positive patients (309 HCV RNA positive by PCR) using a synthetic core peptide-based enzyme immunoassay and PCR amplification of core region sequences with type-specific primers, respectively. Serologic responses to HCV serotypes were successfully identified in 164 (45%) patients, of whom 153 were viremic. Eighty-nine patients had evidence of exposure to serotype 1: 8 of these were infected with genotype I, 50 were infected with genotype II, 2 were infected with genotype III, 7 were infected with genotype V, 13 had infections with mixed genotypes, 3 were infected with an indeterminate genotype, and 6 were nonviremic. Seventy-four patients had been exposed to serotype 2: 64 were infected with genotype III, 3 were infected with mixed genotypes, 2 were infected with an indeterminate genotype, and 5 were nonviremic. The serum of one patient, infected with genotype III, showed reactivity to both serotypes. Comparative evaluation of HCV core region serotyping and genotyping with sera from 294 viremic patients infected with a known HCV genotype showed a remarkable concordance between HCV core region genotyping and serotyping, with only 2 apparently discordant serum samples (both from patients with genotype III infection) of 148 (1.4%) successfully serotyped samples. Serotype 1 infection was more frequently observed in patients with overt chronic liver disease and accounted for all successfully serotyped samples from intravenous drug abusers. In contrast, serotype 2 was more prevalent in subjects with biochemically silent HCV infection (alanine aminotransferase, < 45 U/liter), in agreement with previous findings at the molecular level. HCV core serologic typing is a simple, inexpensive, and highly reproducible assay that can be applied to more than 50% of viremic HCV antibody carriers prior to the use of more sophisticated molecular typing techniques. Moreover, it may be helpful in tracking transmissions routes, particularly for incorrectly stored samples in which the RNA has degraded or for subjects who have cleared the virus and therefore have only antibodies remaining to testify to a remote infection. The lack of recognition of the core sequence from residues 67 to 81, which contains a minor B-cell epitope used to detect type-specific immunoreactivity, may explain the negative serologic findings for half of the patients.


Evaluation of the Chemistry Knowledge of Students Entering the ETH Zurich with a Moodle Quiz.
The basic chemistry knowledge of first-year students in the disciplines Chemistry, Biology, Pharmaceutical Sciences, and Health Sciences and Technology has been evaluated within the first three weeks of the Fall semester with a Moodle-based quiz at ETH Zurich. It consists of 37 small problems testing the knowledge that ETH students entering the lecture courses General Chemistry 1 (Inorganic Chemistry) (AC1) and Organic Chemistry 1 (OC1) should ideally have. An initial set of questions was developed by Bernhard Jaun (ETH Zurich) in 2007, it was combined with questions from an evaluation created in 2015 by Markus Müller (secondary school II teacher). The results of a total of 925 students who took part in the 2016 and 2017 evaluations are presented. It was found that 80% of the students of Chemistry, Chemical Engineering and Interdisciplinary Natural Sciences (AC1 course) and 70% of the students of Biology, Pharmaceutical Sciences, and Health Sciences and Technology (OC1 course) scored ≥60%. Students who took the focus course (Schwerpunktfach) Biology and Chemistry at the SEK II level (Swiss school system) performed on average 13-18% better and with a smaller standard deviation than other students. No significant differences were observed with regard to gender or the region in which the qualification for university entrance was obtained.


Microleakage of three resin placement techniques.
The microleakage associated with three composite resin placement techniques was compared using Class V preparations that extended just beyond the cemento-enamel junction of extracted human molars. The placement techniques were evaluated in conjunction with four adhesive bonding agents. The restored teeth were stored in tap water for 24 hours and then thermally stressed for 100 temperature cycles. Marginal leakage was assessed by measuring the linear penetration of a dye solution. Dye penetration was minimized by using an incremental placement procedure in which the first increment was placed in the coronal portion of the cavity, extended down to the gingivo-axial line angle, and cured. The second increment was used to fill the remainder of the preparation.


Staged Prone/Supine Fixation of High-Energy Multicolumnar Tibial Plateau Fractures: A Multicenter Analysis.
We present a surgical strategy to manage multicolumnar tibial plateau fracture variants by addressing the predominant posterior fragment employing a Lobenhoffer approach in the prone position followed by supine patient repositioning for anterolateral column access. Multicenter retrospective analysis. Three academic Level 1 trauma centers. Twenty-eight cases (28 patients/28 knees) met inclusion criteria between 2003 and 2014. Patient demographic information was retrospectively reviewed with a mean follow-up time of 16.6 months (range 12-34 months). Postoperative radiographic analysis, physical examination findings, and patient outcome scores from the Knee Injury and Osteoarthritis Outcome Score questionnaire were recorded. The average time to union was 3.6 months (range 3-9 months). Eighty-two percent of patients had satisfactory articular reduction (less than 2 mm articular step off). All patients demonstrated satisfactory coronal (medial proximal tibia angle 87 ± 5 degrees) and sagittal alignment (posterior proximal tibia angle 9 ± 4 degrees). Condylar width averaged 2.2 mm. Twenty percent of cases required posterior lateral columnar plating (in addition to posterior medial columnar plating), with none of these cases requiring an extensile exposure modification (medial gastrocnemius origin detachment) to expose posterior laterally. In 12 cases, the posterior approach was staged to allow for anterior soft tissue recovery before subsequent staged supine positioning and lateral column fixation. The knee range of motion averaged 123 degrees (ranged from 2 degrees of extension to 125 degrees flexion). The average Knee Injury and Osteoarthritis Outcome Score was 78/100 (range 29-95). Eleven percent of the patients in the series developed a surgical site infection (n = 3) with 2 requiring formal irrigation and debridement. The most common aseptic complication was radiographic posttraumatic arthrosis (18%). Clinically, 1 patient eventually required a total knee arthroplasty. High-energy multicolumnar tibial plateau fractures with significant posterior articular surface involvement may be predictably addressed with prone positioning, exposure, and fixation followed by supine repositioning and the inclusion of an anterior approach. This study demonstrates excellent postoperative radiographic results and acceptable clinical outcomes resulting from the described staged protocol. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Matrix metalloprotease 2-mediated activation of Ca(2+)-ATPase by superoxide radical (O2*-) in plasma membrane of bovine pulmonary vascular smooth muscle.
The role of the matrix metalloprotease-2 (MMP-2) in regulating Ca(2+)-ATPase activity in bovine pulmonary artery smooth muscle plasma membranes during treatment with the O2*- generating system, hypoxanthine (HPX) plus xanthine oxidase (XO) has been studied. The smooth muscle membranes possess matrix metalloprotease (MMP) activity in gelatin zymogram, having an apparent molecular mass of 72 kDa; the activity is inhibited by the tissue inhibitor of metalloprotease-2 (TIMP-2). Since both protease and MMP-2 have same molecular mass and are inhibited by TIMP-2, it may, therefore, be suggested that the protease is the MMP-2. Treatment of the smooth muscle membrane suspension with the O2*- generating system stimulates MMP-2 activity, as evidenced by an apparent increase in the intensity of the protease activity. O2*- also enhances [14C]-gelatin degradation and Ca(2+)-ATPase activity. The increase in MMP activity, assessed by [14C]-gelatin degradation and Ca(2+)-ATPase activity are inhibited upon pretreatment with superoxide dismutase (SOD). The O2*- triggered MMP and Ca(2+)-ATPase activities in the membrane are found to be inhibited by TIMP-2. The stimulation of the MMP and Ca(2+)-ATPase activities remain unaffected by the inhibitors of serine, thiol and cysteine groups of proteases such as phenylmethylsulfonylfluoride (PMSF), Bowman Birk inhibitor (BBI), chymostatin, N-ethylmaleimide, leupeptin, antipain and pepstatin. Adding pure bovine MMP-2 to the smooth muscle membrane suspension causes an increase in Ca(2+)-ATPase activity, but the pretreatment with TIMP-2 inhibits the increase in the enzyme activity.


Metabolic plasticity for subcutaneous fat accumulation in a long-distance migratory bird traced by 2H2O.
The migrant black-tailed godwit (Limosa limosa) traditionally used natural wetlands in the Iberian Peninsula to prepare for migratory flights by feeding mainly in estuaries. In recent decades, this species has become increasingly dependent on rice fields, thereby relying on a plant-based diet for fuelling. Dietary fatty acids (FA) seem to be determinant to the composition of accumulated subcutaneous fat in migratory birds. It is still unclear whether metabolic plasticity allows for modification and/or synthesis of FA, contributing to a lipid profile that enables a successful migratory performance. Deuterated water was administered to captive black-tailed godwits submitted to two diets (fly larvae versus rice) and the incorporation of deuterium (2H) into subcutaneous triglycerides was analyzed by NMR. A recently developed localized biopsy method for sampling subcutaneous fat was employed with later successful release of all birds into the wild. The average chemical structure reflected mostly a mixture of saturated and monounsaturated 16- and 18-carbon FA, a profile frequently found in migrant birds. Significantly higher levels of polyunsaturated FA, as well as detectable levels of n-3 FA, were observed in fly-larvae-fed birds. Excess 2H-enrichments in FA revealed significantly higher rates of fractional de novo lipogenesis and FA desaturation capacity in rice-fed birds. This novel and non-lethal tracer method revealed the capacity of this species to alter its lipid metabolism to compensate for a poorer dietary lipid contribution. Because of its versatility, adapting this method to other scenarios and/or other migratory species is considered feasible and cost-effective.


Imaginal Retraining Reduces Craving for Tobacco in 1-Year Controlled Follow-Up Study.
Imaginal retraining is a self-help technique that adapts the principles of approach bias modification to the imagination. Imaginal retraining has been shown to reduce craving and addictive behaviours in 3 randomized controlled trials (RCTs) on problematic alcohol consumption, overweight, and tobacco use. To date, there have been no studies evaluating the long-term efficacy of the intervention. The aim of the present study was to generate first hypotheses on the long-term efficacy of imaginal retraining in smokers in a controlled 1-year follow-up study. We recontacted the 345 participants who had taken part in an RCT on imaginal retraining for smokers 1 year later. The survey was carried out online and assessed craving for tobacco (primary outcome), smoking behaviour, well-being, and subjective appraisal. Individuals who applied the technique at least once during the previous year were categorized as the training group, whereas participants who never performed the training were categorized as the no-training group. Data were analysed using linear mixed models (LMMs). The study was preregistered as DRKS00021044. The completion rate was 45.5%. Less than 40% used the intervention at least once in the previous 12 months. LMM analyses showed a significant reduction in craving for tobacco for the training compared to the no-training group after 1 year. No significant group differences emerged in smoking behaviour, depressive symptoms, or quality of life. Subjective appraisal of the intervention was favorable, similar to the initial study. The present study provides preliminary support for the long-term efficacy of imaginal retraining on craving for tobacco but not on smoking behaviour, highlighting the importance of multimodal treatment concepts in smoking cessation that target a variety of maintaining factors. Future studies need to investigate the long-term efficacy of the intervention in prospective RCTs that test alternative ways of conveying the technique to improve adherence.


Network theory and SARS: predicting outbreak diversity.
Many infectious diseases spread through populations via the networks formed by physical contacts among individuals. The patterns of these contacts tend to be highly heterogeneous. Traditional "compartmental" modeling in epidemiology, however, assumes that population groups are fully mixed, that is, every individual has an equal chance of spreading the disease to every other. Applications of compartmental models to Severe Acute Respiratory Syndrome (SARS) resulted in estimates of the fundamental quantity called the basic reproductive number R0--the number of new cases of SARS resulting from a single initial case--above one, implying that, without public health intervention, most outbreaks should spark large-scale epidemics. Here we compare these predictions to the early epidemiology of SARS. We apply the methods of contact network epidemiology to illustrate that for a single value of R0, any two outbreaks, even in the same setting, may have very different epidemiological outcomes. We offer quantitative insight into the heterogeneity of SARS outbreaks worldwide, and illustrate the utility of this approach for assessing public health strategies.


Catecholamines in myocardial ischemia. Systemic and cardiac release.
During myocardial ischemia, malignant arrhythmias and acceleration of cell damage may be induced by sympathetic overstimulation of the heart. This stimulation is due to excessive concentrations of catecholamines within the underperfused myocardium, in combination with enhanced myocyte sensitivity to adrenergic stimuli. Various mechanisms may account for local accumulation of catecholamines in the extracellular space of the ischemic but still viable myocardium. In early myocardial infarction, plasma noradrenaline and adrenaline concentrations are enhanced, reflecting increased activity of the whole sympathetic nervous system, rather than local activity in the heart. In uncomplicated infarction, these concentrations are only five times the normal levels at rest, and there are no convincing data that these mildly increased levels of plasma catecholamines directly induce a major deterioration of myocardial function during the ischemic process. Of more importance is the reflex increase in cardiac sympathetic nerve activity that is induced by pain, anxiety, and a fall in cardiac output or arterial blood pressure and that is accompanied by local exocytotic release of noradrenaline from sympathetic nerve endings of the heart. Excessive accumulation of the neurotransmitter, however, is prevented by at least three mechanisms: 1) Released noradrenaline is rapidly removed so long as neuronal catecholamine reuptake is functional. 2) Adenosine accumulating in the ischemic myocardium effectively suppresses exocytotic noradrenaline release by stimulating presynaptic A1-adenosine receptors. 3) Exocytotic catecholamine release ceases when the sympathetic neurons become depleted of adenosine triphosphate since this release mechanism requires high-energy phosphates. However, with progression of ischemia (i.e., greater than 10 minutes), the myocardium is no longer protected against excess adrenergic stimulation since local metabolic release mechanisms become increasingly important. This release, which is independent of both central sympathetic activation and extracellular calcium, occurs in two steps. First, catecholamines escape from their storage vesicles and accumulate in the cytoplasm of the neuron. In the second, rate-limiting step, noradrenaline is transported across the axolemma from the cytoplasm to the interstitial space via the neuronal uptake carrier in reverse of its normal transport direction. As a consequence of this nonexocytotic local metabolic release, extracellular noradrenaline reaches 100-1,000 times its normal plasma concentrations within 30 minutes of ischemia. Concentrations of this magnitude are capable of producing myocardial necrosis, even in the nonischemic heart, and may play an important role in the pathogenesis of ventricular fibrillation in early ischemia.


The effectiveness of text messages support for diabetes self-management: protocol of the TEXT4DSM study in the democratic Republic of Congo, Cambodia and the Philippines.
People with diabetes find it difficult to sustain adequate self-management behaviour. Self-Management Support strategies, including the use of mobile technology, have shown potential benefit. This study evaluates the effectiveness of a mobile phone support intervention on top of an existing strategy in three countries, DR Congo, Cambodia and the Philippines to improve health outcomes, access to care and enablement of people with diabetes, with 480 people with diabetes in each country who are randomised to either standard support or to the intervention. The study consists of three sub-studies with a similar design in three countries to be independently implemented and analysed. The design is a two-arm Randomised Controlled Trial, in which a total of 480 adults with diabetes participating in an existing DSME programme will be randomly allocated to either usual care in the existing programme or to usual care plus a mobile phone self-management support intervention. Participants in both arms complete assessments at baseline, one year and two years after inclusion.Glycosylated haemoglobin blood pressure, height, weight, waist circumference will be measured. Individual interviews will be conducted to determine the patients' assessment of chronic illness care, degree of self-enablement, and access to care before implementation of the intervention, at intermediate moments and at the end of the study.Analyses of quantitative data including assessment of differences in changes in outcomes between the intervention and usual care group will be done. A probability of <0.05 is considered statistically significant. Outcome indicators will be plotted over time. All data are analysed for confounding and interaction in multivariate regression analyses taking potential clustering effects into account.Differences in outcome measures will be analysed per country and realistic evaluation to assess processes and context factors that influence implementation in order to understand why it works, for whom, under which circumstances. A costing study will be performed. The intervention addresses the problem that the greater part of diabetes management takes place without external support and that many challenges, unforeseen problems and questions occur at moments in between scheduled contacts with the support system, by exploiting communication technology. ISRCTN86247213.


Cough associated with angiotensin-converting enzyme inhibitors.
Cough is reported by between 0.7% and 14% of patients taking angiotensin-converting enzyme (ACE) inhibitors. There are more reports from women and non-smokers. The development of cough during ACE inhibitor therapy is associated with a reversible increase in the response of the cough reflex. The response remains normal in those who do not develop cough while on ACE inhibitor therapy. Both hypersensitivity of the cough reflex and the symptom of cough are reversed by sulindac which suggests that the abnormal reflex is dependent on cyclo-oxygenase products.


Real-time Assessment of the Bidirectional Relationship Between Affective States and Glucose: Protocol for a 14-Day Observational Study.
Glucose variability increases cardiometabolic disease risk. While many factors can influence glucose levels, postprandial glucose response is the primary driver of glucose variability. Furthermore, affect may directly and indirectly impact glucose variability through its effect on eating behavior. Continuous glucose monitors (CGMs) facilitate the real-time evaluation of blood glucose, and ecological momentary assessment (EMA) can be used to assess affect in real time. Together, data collected from these sources provide the opportunity to further understand the role of affect in glucose levels. This paper presents the protocol for a study that aims to (1) evaluate the feasibility and acceptability of using CGMs along with EMA in nondiabetic populations and (2) examine the bidirectional relationship between affect and glucose in nondiabetic adults with overweight or obesity using a CGM and EMA. Eligibility criteria for the study include participants (1) aged 18 to 65 years old, (2) with a BMI of ≥25 kg/m2, (3) who are able to read and write in English, and (4) who own a smartphone. Individuals will be excluded if they (1) have type 1 or 2 diabetes or have any other condition that requires glucose monitoring, (2) are pregnant, (3) use any medications that have the potential to alter blood glucose levels or interfere with the glucose sensing process, or (4) have a diagnosed gastrointestinal condition or eating disorder. In a 14-day observational study, participants will wear a FreeStyle Libre Pro CGM sensor (Abbott) and will receive mobile phone-based EMA prompts 6 times per day (randomly within six 2-hour windows between 8 AM and 8 PM) to assess positive and negative affect. Participants will also wear a Fitbit Inspire 2 (Fitbit) to continuously monitor physical activity and sleep, which will be included as covariates in the analysis. Multilevel linear regression models will be used to evaluate the acute relationship between glucose level and affect. Recruitment started in October 2022 and is expected to be completed in March 2023. We will aim to recruit 100 participants. As of December 12, 2022, a total of 39 participants have been enrolled. The results of this study will further elucidate the role of affect in glucose variability. By identifying affective states that may lead to glucose excursions, our findings could inform just-in-time behavioral interventions by indicating opportunities for intervention delivery. DERR1-10.2196/45104.


Measurement of the pesticide methomyl by modified quartz crystal nanobalance with molecularly imprinted polymer.
A simple and cost-effective analysis method based on quartz crystal nanobalance (QCN) coated with a molecularly imprinted polymer (MIP) for measurement of methomyl was investigated. In the first part of this study, a sensitive, selective and reliable quartz crystal nanobalance (QCN) sensor was designed for the selective determination of methomyl in aqueous solutions. In the second part, in order to demonstrate the applicability and performance of the fabricated sensor in the real world situation, it was successfully applied for the determination of methomyl residual in photo catalytic degradation by ZnO powders in aqueous solutions. The fabricated sensor presents a high selectivity and sensitivity (4.56 Hz per mg L(-1)) for methomyl and it can be used for determination of methomyl concentration ranged between 1 to 45 mg L(-1). Furthermore, good reproducibility, R.S.D. = 2.14% (n = 5) was observed. To investigate the performance of the sensor, the change in the insecticide concentration during the photocatalytic degradation of methomyl by ZnO was investigated by QCN and UV/Vis spectroscopy. Results obtained from QCN sensor and UV/Vis spectroscopy measurement are in good mutual agreement. So the fabricated sensor may provide an efficient, low cost, easy-to-use method for the in-field evaluation of specific targeted analytes in aqueous solutions which in turn may lead to improved food and water safety.


TRAK App Suite: A Web-Based Intervention for Delivering Standard Care for the Rehabilitation of Knee Conditions.
Standard care for the rehabilitation of knee conditions involves exercise programs and information provision. Current methods of rehabilitation delivery struggle to keep up with large volumes of patients and the length of treatment required to maximize the recovery. Therefore, the development of novel interventions to support self-management is strongly recommended. Such interventions need to include information provision, goal setting, monitoring, feedback, and support groups, but the most effective methods of their delivery are poorly understood. The Internet provides a medium for intervention delivery with considerable potential for meeting these needs. The objective of this study was to demonstrate the feasibility of a Web-based app and to conduct a preliminary review of its practicability as part of a complex medical intervention in the rehabilitation of knee disorders. This paper describes the development, implementation, and usability of such an app. An interdisciplinary team of health care professionals and researchers, computer scientists, and app developers developed the TRAK app suite. The key functionality of the app includes information provision, a three-step exercise program based on a standard care for the rehabilitation of knee conditions, self-monitoring with visual feedback, and a virtual support group. There were two types of stakeholders (patients and physiotherapists) that were recruited for the usability study. The usability questionnaire was used to collect both qualitative and quantitative information on computer and Internet usage, task completion, and subjective user preferences. A total of 16 patients and 15 physiotherapists participated in the usability study. Based on the System Usability Scale, the TRAK app has higher perceived usability than 70% of systems. Both patients and physiotherapists agreed that the given Web-based approach would facilitate communication, provide information, help recall information, improve understanding, enable exercise progression, and support self-management in general. The Web app was found to be easy to use and user satisfaction was very high. The TRAK app suite can be accessed at http://apps.facebook.com/kneetrak/. The usability study suggests that a Web-based intervention is feasible and acceptable in supporting self-management of knee conditions.


Bidirectional activity-dependent plasticity of membrane potential and the influence on spiking in rat hippocampal dentate granule cells.
Granule cells of the dentate gyrus in the hippocampus generally fire at low frequencies but are known to respond to sensory cues by increasing their rate of firing. We have previously shown that a burst of action potentials in synaptically isolated granule cells can induce a long-term depolarisation (LTDepol) of the neuronal membrane potential. This form of excitability plasticity could be an important mechanism for learning and memory. Here we demonstrate that this depolarisation can be reversed by physiologically relevant firing patterns. At a basal action potential frequency of 0.1Hz the membrane potential depolarises in response to brief high frequency stimulation (HFS) but this depolarisation is blocked or reversed by 1Hz action potential firing. The depolarisation of the neurones did not, however, affect the input-output function of the dentate gyrus measured by field or single cell recordings. HFS or brief forskolin application that mimics LTDepol did not alter the excitatory input-output spike coupling measured using field potential recordings. Similarly, LTDepol did not change the probability of spike firing in response to a given input. The mechanism underlying the lack of change in input-output relationship was found to be a concurrent depolarisation of the threshold potential for the initiation of action potentials. We demonstrate that physiologically relevant patterns of activity regulate the membrane potential of dentate gyrus granule cells in a bidirectional manner but the changes in membrane potential do not alter the excitability of these neurones measured by their response to excitatory inputs.


Roles and regulation of membrane-associated serine proteases.
Pericellular proteolytic activity affects many aspects of cellular behaviour, via mechanisms involving processing of the extracellular matrix, growth factors and receptors. The serine proteases have exquisitely sensitive regulatory mechanisms in this setting, involving both receptor-bound and transmembrane proteases. Receptor-bound proteases are exemplified by the uPA (urokinase plasminogen activator)/uPAR (uPAR receptor) plasminogen activation system. The mechanisms initiating the activity of this proteolytic system on the cell surface, a critical regulatory point, are poorly understood. We have found that the expression of the TTSP (type II transmembrane serine protease) matriptase is highly regulated in leucocytes, and correlates with the presence of active uPA on their surface. Using siRNA (small interfering RNA), we have demonstrated that matriptase specifically activates uPAR-associated pro-uPA. The uPA/uPAR system has been implicated in the activation of the plasminogen-related growth factor HGF (hepatocyte growth factor). However, we find no evidence for this, but instead that HGF can be activated by both matriptase and the related TTSP hepsin in purified systems. Hepsin is of particular interest, as the proteolytic cleavage sequence of HGF is an 'ideal substrate' for hepsin and membrane-associated hepsin activates HGF with high efficiency. Both of these TTSPs can be activated autocatalytically at the cell surface, an unusual mechanism among the serine proteases. Therefore these TTSPs have the capacity to be true upstream initiators of proteolytic activity with subsequent downstream effects on cell behaviour.