Molecular and antigenic characterization of rabies viruses from Iran identifies variants with distinct epidemiological origins.
A molecular epidemiological study of 48 recent rabies isolates recovered from cases reported throughout Iran identified three distinct viral variants, the evolutionary origins of which were identified by phylogenetic comparison with rabies viruses originating from Europe and Asia. Members of group 1 (15 isolates) were recovered from the northern half of the country only, while those of group 2 (31 isolates) were widely dispersed; both groups clustered within the widely disseminated cosmopolitan lineage. The two isolates of group 3 were detected in the northeastern tip of the country only and belonged to the Arctic strain. Rapid variant discrimination tools, employing restriction fragment length polymorphisms applied to amplified fragments of the viral genome, were devised whilst antigenic characterization of representative viruses identified a small panel of monoclonal antibodies that were also discriminatory. The future application of such methods should provide valuable epidemiological information on rabies incidence in Iran.


Effects of Microbial Aerosol in Poultry House on Meat Ducks' Immune Function.
The aim of this study was to evaluate effects of microbial aerosols on immune function of ducks and shed light on the establishment of microbial aerosol concentration standards for poultry. A total of 1800 1-d-old cherry valley ducks were randomly divided into five groups (A, B, C, D, and E) with 360 ducks in each. To obtain objective data, each group had three replications. Concentrations of airborne bacteria, fungi, endotoxin in different groups were created by controlling ventilation and bedding cleaning frequency. Group A was the control group and hygienic conditions deteriorated progressively from group B to E. A 6-stage Andersen impactor was used to detect the aerosol concentration of aerobes, gram-negative bacteria, fungi, and AGI-30 microbial air sampler detect the endotoxin, and Composite Gas Detector detect the noxious gas. In order to assess the immune function of meat ducks, immune indicators including H5 AIV antibody titer, IgG, IL-2, T-lymphocyte transformation rate, lysozyme and immune organ indexes were evaluated. Correlation coefficients were also calculated to evaluate the relationships among airborne bacteria, fungi, endotoxin, and immune indicators. The results showed that the concentration of airborne aerobe, gram-negative bacteria, fungi, endotoxin have a strong correlation to H5 AIV antibody titer, IgG, IL-2, T-lymphocyte transformation rate, lysozyme, and immune organ indexes, respectively. In addition, when the concentration of microbial aerosol reach the level of group D, serum IgG (6-8 weeks), lysozyme (4 week) were significantly higher than in group A (P < 0.05); serum IL-2 (7 and 8 weeks), T-lymphocyte transformation rate, lysozyme (7 and 8 weeks), spleen index (6 and 8 weeks), and bursa index (8 week) were significantly lower than in group A (P < 0.05 or P < 0.01). The results indicated that a high level of microbial aerosol adversely affected the immune level of meat ducks. The microbial aerosol values in group D provide a basis for recommending upper limit concentrations of microbial aerosols for healthy meat ducks.


Food hoarding is increased by pregnancy, lactation, and food deprivation in Siberian hamsters.
Food hoarding by male Siberian hamsters (Phodopus sungorus sungorus) is increased only when body mass (fat) is decreased. Pregnancy and lactation result in marked decreases in lipid reserves (approximately 50%) in female Siberian hamsters. Therefore, the present experiments addressed the following questions: 1) Is food hoarding increased after food deprivation in female Siberian hamsters? and 2) How do food hoarding and food intake change during pregnancy, lactation, and their combination? During measurements in a simulated burrow system food hoarding increased after a 32-h fast (approximately 2- to 3-fold) to a level similar to that seen previously in males and was markedly increased during pregnancy (approximately 12- to 18-fold, lactation, and concurrent pregnancy and lactation (approximately 10- to 25-fold for each of the latter 2 conditions). Postfast food intake was not different from prefast baseline measures. Food intake was increased only during the last few days of pregnancy and was elevated throughout lactation. These impressive increases in the level of food hoarding during pregnancy, lactation and their combination suggest that food hoarding may play an important role in supplying easily accessible energy to subserve these reproductive conditions.


Influence of Active Layer on Separation Potentials of Nanofiltration Membranes for Inorganic Ions.
Active layers of two fully aromatic and two semi-aromatic nanofiltration membranes were studied along with surface charge at different electrolyte composition and effective pore size to elucidate their influence on separation mechanisms for inorganic ions by steric, charge, and dielectric exclusion. The membrane potential method used for pore size measurement is underlined as the most appropriate measurement technique for this application owing to its dependence on the diffusional potentials of inorganic ions. Crossflow rejection experiments with dilute feed composition indicate that both fully aromatic membranes achieved similar rejection despite the differences in surface charge, which suggests that rejection by these membranes is exclusively dependent on size exclusion and the contribution of charge exclusion is weak. Rejection experiments with higher ionic strength and different composition of the feed solution confirmed this hypothesis. On the other hand, increase in the ionic strength of feed solution when the charge exclusion effects are negligible due to charge screening strongly influenced ion rejection by semi-aromatic membranes. The experimental results confirmed that charge exclusion contributes significantly to the performance of semi-aromatic membranes in addition to size exclusion. The contribution of dielectric exclusion to overall ion rejection would be more significant for fully aromatic membranes.


Primary sequence and secondary structure motifs in spleen necrosis virus RU5 confer translational utilization of unspliced human immunodeficiency virus type 1 reporter RNA.
The 5' long terminal repeat (LTR) of spleen necrosis virus (SNV) contains a unique posttranscriptional control element that facilitates Rev/Rev-responsive element-independent expression of unspliced human immunodeficiency virus type 1 (HIV-1) gag reporter RNA. HIV-1 Gag expression is eliminated when SNV LTR is repositioned to the 3' untranslated region or when the RU5 region is positioned in the antisense orientation. RU5 corresponds to the 5' RNA terminus, and results presented here indicate that Gag production is sustained upon introduction of transcribed spacers that reposition SNV RU5 35 to 200 nucleotides downstream. Concordant results of deletion and point mutagenesis identified two functionally redundant and synergistic motifs (designated A and C) that are necessary and sufficient for SNV RU5 activity. Enzymatic analysis of SNV RU5 RNA structure determined that A and C correspond to stem-loop structures. Quantitative RNA and protein analysis of A and C mutants revealed that the structural integrity of A and C is necessary for protein production, and loss of function correlates with little change in steady-state level, splicing efficiency, or cytoplasmic accumulation of HIV-1 gag reporter RNA. Instead, the structural mutations eliminate cytoplasmic utilization as an mRNA template for Gag protein production. Point mutations of unpaired loop-and-bulge nucleotides that maintain the structure of A eliminate activity. The results show that the unpaired UUGU loop and U-rich bulges function together and are candidate SNV RU5 binding sites for the host cell protein(s) that directs cytoplasmic utilization of unspliced HIV-1 reporter RNA.


Extracellular vesicles from osteosarcoma cell lines contain miRNAs associated with cell adhesion and apoptosis.
Osteosarcoma is the most common primary bone tumor during childhood and adolescence. Several reports have presented data on serum biomarkers for osteosarcoma, but few reports have analyzed circulating microRNAs (miRNAs). In this study, we used next generation miRNA sequencing to examine miRNAs isolated from microvesicle-depleted extracellular vesicles (EVs) derived from six different human osteosarcoma or osteoblastic cell lines with different degrees of metastatic potential (i.e., SAOS2, MG63, HOS, 143B, U2OS and hFOB1.19). EVs from each cell line contain on average ~300 miRNAs, and ~70 of these miRNAs are present at very high levels (i.e., >1000 reads per million). The most prominent miRNAs are miR-21-5p, miR-143-3p, miR-148a-3p and 181a-5p, which are enriched between 3 and 100 fold and relatively abundant in EVs derived from metastatic SAOS2 cells compared to non-metastatic MG63 cells. Gene ontology analysis of predicted targets reveals that miRNAs present in EVs may regulate the metastatic potential of osteosarcoma cell lines by potentially inhibiting a network of genes (e.g., MAPK1, NRAS, FRS2, PRCKE, BCL2 and QKI) involved in apoptosis and/or cell adhesion. Our data indicate that osteosarcoma cell lines may selectively package miRNAs as molecular cargo of EVs that could function as paracrine agents to modulate the tumor micro-environment.


Enhanced autophagy contributes to protective effects of IL-22 against acetaminophen-induced liver injury.
Acute or acute-on-chronic liver failure is a leading cause of death in liver diseases without effective treatment. Interleukin-22 (IL-22) is currently in clinical trials for the treatment of severe alcoholic hepatitis, but the underlying mechanisms remain to be explored. Autophagy plays a critical role in alleviating liver injury. The aim of the current study is to explore the role of autophagy in IL-22-mediated hepato-protective effect against acetaminophen (APAP)-induced liver injury. Methods: A model of acute liver injury induced by APAP was used in vivo. IL-22 was administrated to the APAP-treated mice. Hepatocytes were pre-incubated with IL-22, followed by exposure to APAP for in vitro analyses. Results: IL-22 administration significantly reduced serum ALT and AST, hepatic reactive oxygen species, and liver necrosis in APAP-challenged mice. APAP treatment increased hepatic autophagosomes, which was further intensified by IL-22 co-treatment. Hepatic LC3-II was moderately upregulated after APAP administration without obvious alteration of phosphorylation of AMP-activated kinase (p-AMPK). IL-22 pretreatment significantly upregulated hepatic LC3-II and p-AMPK in APAP-treated mice. IL-22 also alleviated APAP-induced cytotoxicity and upregulated LC3-II and p-AMPK expression in cultured hepatocytes treated with APAP in vitro. When p-AMPK was blocked with compound C (an AMPK inhibitor), IL-22-mediated LC3-II conversion and protection against APAP-induced cytotoxicity was weakened. Conclusions: Enhanced AMPK-dependent autophagy contributes to protective effects of IL-22 against APAP-induced liver injury.


DNA methylation age of blood predicts all-cause mortality in later life.
DNA methylation levels change with age. Recent studies have identified biomarkers of chronological age based on DNA methylation levels. It is not yet known whether DNA methylation age captures aspects of biological age. Here we test whether differences between people's chronological ages and estimated ages, DNA methylation age, predict all-cause mortality in later life. The difference between DNA methylation age and chronological age (Δage) was calculated in four longitudinal cohorts of older people. Meta-analysis of proportional hazards models from the four cohorts was used to determine the association between Δage and mortality. A 5-year higher Δage is associated with a 21% higher mortality risk, adjusting for age and sex. After further adjustments for childhood IQ, education, social class, hypertension, diabetes, cardiovascular disease, and APOE e4 status, there is a 16% increased mortality risk for those with a 5-year higher Δage. A pedigree-based heritability analysis of Δage was conducted in a separate cohort. The heritability of Δage was 0.43. DNA methylation-derived measures of accelerated aging are heritable traits that predict mortality independently of health status, lifestyle factors, and known genetic factors.


The integration of a Podiatrist into an orthopaedic department: a cost-consequences analysis.
The aim of this study was to evaluate the cost-consequences of a podiatry-led triage clinic provided in an orthopaedic department relative to usual care for non-urgent foot and ankle complaints in an Australian tertiary care hospital. All new, non-urgent foot and ankle patients seen in an outpatient orthopaedic department were included in this study. The patients seen between 2014 and 2015 by Orthopaedic Surgeons were considered 'usual care', the patients seen between 2015 and 2016 by a Podiatrist were considered the 'Podiatry Triage Clinic'. Data on new and review patient appointments; the number of new patients / session; the number of appointments / patient; the number of patients discharged; the surgical conversion rate; staff time; and imaging use were collected. A cost-consequences analysis, undertaken from a healthcare provider perspective (hospital) estimated the incremental resource use, costs and effects of the Podiatry Triage Clinic relative to usual care over a 12-month period. The Orthopaedic Surgeons and Podiatrist consulted with 72 and 212 new patients during the usual care and triage periods, respectively. The Podiatrist consulted with more new patients / session, mean (SD) of 3.6 (1.0) versus 0.7 (0.8), p < 0.001 and utilised less appointments / patient than the Orthopaedic Surgeons, mean (SD) of 1.3 (0.6) versus 1.9 (1.1), p < 0.001. The percentage of patients discharged without surgery was similar in the Podiatry Triage Clinic and usual care, 80.3% and 87.5% p = 0.135, respectively, but the surgical conversion rate was higher in the Podiatry Triage Clinic, 76.1% versus 12.5% p < 0.001. The total integrated appointment cost for the 12-month usual care period was $32,744, which represented a cost of $454.78 / patient. The total appointment and imaging cost during the triage period was $19,999, representing $94.34 / patient. Further analysis, suggests that the projected annual saving of integrating a Podiatry Triage Clinic versus an orthopaedic clinic alone is $50,441. The integration of a Podiatrist into an orthopaedic department significantly increases the number of patients seen, is cost-effective, improves the surgical conversion rate and improves the utilisation of Orthopaedic Surgeons.


Dilute random distribution of small chiral spheres.
The problem solved here consists of estimating the effective-medium properties of a chiral composite made of a dilute concentration of small noninteracting chiral spheres, randomly suspended in free space. Volume integral equations, to determine the scattering characteristics of an inhomogeneous chiral scatterer, are obtained. These equations are used to derive the general electromagnetic polarizability matrix of a small, homogeneous, chiral sphere embedded in free space. Finally, from the polarizability matrix, several conclusions regarding the effective properties of the chiral composite are obtained.


Mild chronic stress leads to desensitisation of presynaptic autoreceptors and a long-lasting increase in noradrenaline synthesis in rat cortical synaptosomes.
An i.p. injection of normal saline combined with 1 min handling when repeated 14 times results in an increase in noradrenaline synthesis in synaptosomes prepared from the cortex of stressed rats; at 24 h synthesis acceleration is greater than at 48 h after the last stress. The activity of tyrosine hydroxylase solubilised from the hippocampus is the same in the control and the stressed group, when assayed at the optimal pH of 5.8 and with saturating concentration (2 mM) of the cofactor 6 MPH(4). However enzyme from stressed rats shows a relative increase in the activity at pH 7.4 assayed in the presence of 0.2 mM 6 MPH(4). This indicates activation, not induction, of the enzyme. 8-Br-cAMP produced the same increase in noradrenaline synthesis in cortical synaptosomes from control and stressed rats; however 50 mM K(+) did not increase synthesis rate in stressed rats. Furthermore in synaptosomes from stressed rats neither isoprenaline (which increases noradrenaline synthesis) nor clonidine with 50 mM K(+) (which leads to a depression of the K(+)-accelerated synthesis) had any effect on synthesis rate. The results suggest that the increased noradrenaline synthesis rate in cortical synaptosomes from stressed rats represents a Ca(2+)-dependent activation of tyrosine hydroxylase resulting from the desensitisation of alpha(2)-autoreceptors.


An interdisciplinary approach to inhospital stroke improves stroke detection and treatment time.
Inhospital stroke (IHS) is associated with high morbidity and mortality, likely related to multiple factors, including delayed time to recognition, associated comorbidities, and initial care from non-stroke trained providers. We hypothesized that guided revision of a formalized 'stroke code' system can improve diagnosis and time to thrombolysis and thrombectomy. IHS activations occurring at a comprehensive stroke center between 2013 and 2016 were retrospectively analyzed to guide revisions of an established stroke code protocol to improve provider communication and time to imaging, reduce stroke mimic rate, and improve the use of parallel processing. After protocol implementation, we prospectively collected data between 2016 and 2017 for comparison with the pre-implementation group, including diagnostic accuracy and relevant time points (code call to examination, examination to imaging, and imaging to intervention). We report descriptive statistics for comparison of patient characteristics and time metrics (time to imaging and reperfusion after IHS activation). Multivariable regression analysis was performed to identify independent predictors of stroke mimics and time metrics. There were 136 cases in the pre-implementation group and 69 in the post-implementation group. A reduction in stroke mimics (52% vs 33%, P=0.01) occurred after protocol initiation. Mean time to imaging after stroke code call was 7.6 min shorter (P=0.026) and mean time from imaging to acute reperfusion therapy was 45.7 vs 19.8 min (P=0.05) in the pre- versus the post-implementation group. Revision of an existing IHS protocol was associated with a lower rate of stroke mimics, and a shorter time to intravenous and intra-arterial intervention.


Making a gender difference: case studies of gender mainstreaming in medical education.
Curricula are accommodated to the interests of new groups after pressure from social movements outside institutions. A Dutch national project to integrate gender-gender mainstreaming (GM)-in all medical curricula started in 2002 and finished in 2005. GM is a long-term strategy which aims at eliminating gender bias in existing routines for which involvement of regular actors within the organization is required. In this paper, the challenges of GM in medical education are discussed. Three case studies of medical schools are presented to identify key issues in the change process. Steps taken in the national project included the evaluation of a local project, establishing a digital knowledge centre with education material, involving stakeholders and building political support within the schools and national bodies, screening education material and negotiating recommendations with course organizers, and evaluating the project with education directors and change agents. Data are gathered from interviews and document analysis. Factors playing a role are distinguished at three levels: (1) policy level, such as political support and widespread communication of this support; (2) organizational level such as a problem-based curricula and procedures for curriculum development and evaluation; and (3) faculty's openness towards change in general and towards feminist influences in particular, and change agents' position as well as personal and communicative skills. Successful GM in medical education is both a matter of strategy as well as how such strategy is received in medical schools. A time-consuming strategy could overcome resistance as well as dilemmas inherent in GM. More female teachers than male teachers were openly accepting. However, women were situated in less visible and less powerful positions. Hence, GM is accelerated by alliances between women aiming for change and senior (male) faculty leadership.


Prediction of tumor radiosensitivity in rectal carcinoma based on p53 and Ku70 expression.
The identification of predictive indicators of radiosensitivity is extremely useful in selecting patients suited for preoperative radiotherapy and avoiding unnecessary preoperative treatment. In this study, we evaluated the possible role of the immunohistochemical expression pattern of p53 and Ku70 protein in determining tumor radiosensitivity in rectal cancer before preoperative irradiation. We examined pretreatment biopsy materials from 111 patients by immunohistochemistry. The expression pattern of p53 and Ku70 was evaluated for association with tumor radiosensitivity, which was defined according to the criteria of the Japanese Research Society for Cancer of the Colon and Rectum. There was a significant correlation between the expression pattern of p53 and tumor radiosensitivity (P = 0.045); Ku70 and tumor radiosensitivity (P < 0.001); and the combination of p53 and Ku70, and tumor radiosensitivity (P < 0.001). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in both p53 and Ku70-positive cases for radioresistance were all superior to those of the group positive for p53 alone. In conclusion the examination of the combination of p53 and Ku70 may predict the radiosensitivity of rectal cancer before preoperative irradiation.


The application of mass and energy conservation laws in physiologically structured population models of heterotrophic organisms
Rules for energy uptake, and subsequent utilization, form the basis of population dynamics and, therefore, explain the dynamics of the ecosystem structure in terms of changes in standing crops and size distributions of individuals. Mass fluxes are concomitant with energy flows and delineate functional aspects of ecosystems by defining the roles of individuals and populations. The assumption of homeostasis of body components, and an assumption about the general structure of energy budgets, imply that mass fluxes can be written as weighted sums of three organizing energy fluxes with the weight coefficients determined by the conservation law of mass. These energy fluxes are assimilation, maintenance and growth, and provide a theoretical underpinning of the widely applied empirical method of indirect calorimetry, which relates dissipating heat linearly to three mass fluxes: carbon dioxide production, oxygen consumption and N-waste production. A generic approach to the stoichiometry of population energetics from the perspective of the individual organism is proposed and illustrated for heterotrophic organisms. This approach indicates that mass transformations can be identified by accounting for maintenance requirements and overhead costs for the various metabolic processes at the population level. The theoretical background for coupling the dynamics of the structure of communities to nutrient cycles, including the water balance, as well as explicit expressions for the dissipating heat at the population level are obtained based on the conservation law of energy. Specifications of the general theory employ the Dynamic Energy Budget model for individuals. Copyright 1999 Academic Press.


The controlled drug release by pH-sensitive molecularly imprinted nanospheres for enhanced antibacterial activity.
In this study, we prepared pH-sensitive hybrid nanospheres through the implementation of a facile molecularly imprinted polymer (MIP) technique combined with a UV-initiated precipitation polymerization method using vancomycin (VA) for the templates. During the course of this investigation, both 2-hydroxyethyl methacrylate (HEMA) and 2-(diethylamino) ethyl methacrylate (DEAEMA) were utilized as the functional monomers, while ethylene glycol dimethacrylate (EGDMA) was used as a cross-linker. The obtained MIP nanospheres exhibited well-controlled particle size, with a drug loading capacity of about 17%, much higher than that of the non-imprinted polymer (NIP) nanospheres (5%). In addition, the VA loading quantity was closely correlated with the dosage of the cross-linking agent, and the MIP nanospheres exhibited a slower and more controlled VA release rate than the NIP nanospheres. Moreover, these MIP nanospheres were sensitive to pH values, and consequently showed an increasing release rate of VA as the pH level was decreased. The VA-loaded MIP nanospheres showed the higher antibacterial ratio of over 92% against Staphylococcus aureus (S. aureus) while the NIP nanospheres were friendly to S. aureus. These MIP nanospheres can be promising for targeting drug delivery system to achieve specific therapies such as preventing bacterial infections and killing cancer cells without damaging health cells and tissues.


Sequential trans-dimensional Monte Carlo for range-dependent geoacoustic inversion.
This paper develops a sequential trans-dimensional Monte Carlo algorithm for geoacoustic inversion in a strongly range-dependent environment. The algorithm applies advanced Markov chain Monte Carlo methods in combination with sequential techniques (particle filters) to carry out geoacoustic inversions for consecutive data sets acquired along a track. Changes in model parametrization along the track (e.g., number of sediment layers) are accounted for with trans-dimensional partition modeling, which intrinsically determines the amount of structure supported by the data information content. Challenging issues of rapid environmental change between consecutive data sets and high information content (peaked likelihood) are addressed by bridging distributions implemented using annealed importance sampling. This provides an efficient method to locate high-likelihood regions for new data which are distant and ∕ or disjoint from previous high-likelihood regions. The algorithm is applied to simulated reflection-coefficient data along a track, such as can be collected using a towed array close to the seabed. The simulated environment varies rapidly along the track, with changes in the number of layers, layer thicknesses, and geoacoustic parameters within layers. In addition, the seabed contains a geologic fault, where all layers are offset abruptly, and an erosional channel. Changes in noise level are also considered.


What are the most important unanswered research questions in trial retention? A James Lind Alliance Priority Setting Partnership: the PRioRiTy II (Prioritising Retention in Randomised Trials) study.
One of the top three research priorities for the UK clinical trial community is to address the gap in evidence-based approaches to improving participant retention in randomised trials. Despite this, there is little evidence supporting methods to improve retention. This paper reports the PRioRiTy II project, a Priority Setting Partnership (PSP) that identified and prioritised unanswered questions and uncertainties around trial retention in collaboration with key stakeholders. This PSP was conducted in collaboration with the James Lind Alliance, a non-profit making initiative, to support key stakeholders (researchers, patients, and the public) in jointly identifying and agreeing on priority research questions. There were three stages. (1) First an initial online survey was conducted consisting of six open-ended questions about retention in randomised trials. Responses were coded into thematic groups to create a longlist of questions. The longlist of questions was checked against existing evidence to ensure that they had not been answered by existing research. (2) An interim stage involved a further online survey where stakeholders were asked to select questions of key importance from the longlist. (3) A face-to-face consensus meeting was held, where key stakeholder representatives agreed on an ordered list of 21 unanswered research questions for methods of improving retention in randomised trials. A total of 456 respondents yielded 2431 answers to six open-ended questions, from which 372 questions specifically about retention were identified. Further analysis included thematically grouping all data items within answers and merging questions in consultation with the Steering Group. This produced 27 questions for further rating during the interim survey. The top 21 questions from the interim online survey were brought to a face-to-face consensus meeting in which key stakeholder representatives prioritised the order. The 'Top 10' of these are reported in this paper. The number one ranked question was 'What motivates a participant's decision to complete a clinical trial?' The entire list will be available at www.priorityresearch.ie . The Top 10 list can inform the direction of future research on trial methods and be used by funders to guide projects aiming to address and improve retention in randomised trials.


Sex and Diffusion Tensor Imaging of White Matter in Schizophrenia: A Systematic Review Plus Meta-analysis of the Corpus Callosum.
Sex is considered an understudied variable in health research. Schizophrenia is a brain disorder with known sex differences in epidemiology and clinical presentation. We systematically reviewed the literature for sex-based differences of diffusion properties of white matter tracts in schizophrenia. We then conducted a meta-analysis examining sex-based differences in the genu and splenium of the corpus callosum in schizophrenia. Medline and Embase were searched to identify relevant papers. Studies fulfilling the following criteria were included: (1) included individuals with a diagnosis of schizophrenia, (2) included a control group of healthy individuals, (3) included both sexes in the patient and the control groups, (4) used diffusion tensor imaging, and (5) involved analyzing metrics of white matter microstructural integrity. Fractional anisotropy (FA) was used as the measure of interest in the meta-analysis. Of 730 studies reviewed, 75 met the inclusion criteria. Most showed no effect of sex, however, those that did found either that females have lower FA than males, or that the effect of disease in females is larger than that in males. The findings of the meta-analysis in the corpus callosum supported this result. There is a recognized need for studies on schizophrenia with a sufficient sample of female patients. Lack of power undermines the ability to detect sex-based differences. Understanding the sex-specific impact of illness on neural circuits may help inform development of new treatments, and improvement of existing interventions.


N-formyl peptide receptors cluster in an active raft-associated state prior to phosphorylation.
In response to ligand binding, G protein-coupled receptors undergo phosphorylation and activate cellular internalization machinery. An important component of this process is the concentration of receptors into clusters on the plasma membrane. Aside from organizing the receptor in anticipation of internalization, little is known of the function of ligand-mediated G protein-coupled receptor clustering, which has traditionally been thought of as being a phosphorylation-dependent event prior to receptor internalization. We now report that following receptor activation, the N-formyl peptide receptor (FPR) forms distinct membrane clusters prior to its association with arrestin. To determine whether this clustering is dependent upon receptor phosphorylation, we used a mutant form of the FPR, DeltaST-FPR, which lacks all phosphorylation sites in the carboxyl-terminal domain. We found that activation of the signaling-competent DeltaST-FPR resulted in rapid receptor clustering on the plasma membrane independent of Gi protein activation. This clustering required receptor activation since the D71A mutant receptor, which binds ligand but is incapable of transitioning to an active state, failed to induce receptor clustering. Furthermore we demonstrated that FPR-mediated clustering and signaling were cholesterol-dependent processes, suggesting that translocation of the active receptor to lipid rafts may be required for maximal signaling activity. Finally we showed that FPR stimulation in the absence of receptor phosphorylation resulted in translocation of FPR to GM1-rich clusters. Our results demonstrate for the first time that formation of a clustered activated receptor state precedes receptor phosphorylation, arrestin binding, and internalization.


Differentiated and non-differentiated HepaRG™ cells: A possible in-vitro model system for early hepatocarcinogenesis and non-genotoxic carcinogens.
Many xenobiotics are non-genotoxic carcinogens (NGC) in rodent liver. Their mode of action (MoA) and health risks for humans are unclear and no in-vitro tests are available to predict NGC. Human HepaRG™ cells in the differentiated (d-HepaRG) and non-differentiated state (nd-HepaRG) were studied as new approach methodology (NAM) for NGC. Cell-biological assays were performed with d-/nd-HepaRG and human hepatoma/hepatocarcinoma cell lines to characterize the benign/malignant phenotype. Reaction of d-/nd-HepaRG to several liver growth factors and NGC (phenobarbital, PB; cyproterone acetate, CPA; WY-14643) was compared to unaltered and premalignant rat hepatocytes in ex-vivo culture. Enzyme induction by NGC was checked by RT-qPCR/oligo-arrays. Growth, anchorage-independency, migration, clonogenicity, and in-vivo tumorigenicity of nd-HepaRG ranged between benign d-HepaRG and malignant hepatoma/hepatocarcinoma cells. All growth factors elevated DNA replication of d-/nd-HepaRG cells, similarly to unaltered/premalignant rat hepatocytes. NGC induced their prototypical enzymes in the rat and human cells, but elicited a growth response only in the unaltered/premalignant rat hepatocytes and not in human d-/nd-HepaRG cells. To conclude, a benign/premalignant phenotype of d-/nd-HepaRG cells and a reactivity towards several hepatic growth factors and NGC, as known from human hepatocytes, are essential components for an in-vitro model for early stage human hepatocarcinogenesis.The potential value as new approach methodology (NAM) for NGC is discussed.


Socioeconomic status significantly contributes to the likelihood of immediate postmastectomy breast reconstruction in the Netherlands: A nationwide study.
Previous studies have shown that breast cancer patients with a low socioeconomic status (SES) are less likely to undergo postmastectomy immediate breast reconstruction (IBR). However, these studies were performed in countries with unequal access to healthcare. Therefore, the aim of this study was to investigate whether SES also contributes to the likelihood of receiving IBR in a country with equal access to healthcare. Patients with stage I or II breast cancer diagnosed between 2011 and 2018 who underwent mastectomy were selected from the Netherlands Cancer Registry. SES was calculated from the average incomes of each postal code which were divided into 10-deciles. Primary outcome was the effect of SES on the likelihood of receiving IBR, controlled for patient, tumour and hospital characteristics expressed as Odds Ratio (OR) with 95% confidence interval (CI). Higher SES significantly increased the probability of undergoing postmastectomy IBR (OR 1.05 per 10% SES stratum), just as larger hospital volume (average volume OR 1.89 and large volume 2.58), oestrogen positive tumours (OR 1.19) and neo-adjuvant therapy (OR 1.42). In contrast, factors significantly reducing the likelihood of receiving IBR were older age (OR 0.92 per year), stage II (OR 0.61 compared to stage I) and adjuvant therapy (OR 0.56). Women with lower SES undergoing mastectomy were less likely to receive postmastectomy IBR. More research is warranted to study whether lifestyle factors associated with lower SES such as smoking and higher BMI, language barrier, illiteracy and less access to internet explain these differences.


Mycobacterium septicum: a 6-Year Clinical Experience from a Tertiary Hospital and Reference Laboratory.
Mycobacterium septicum is a rarely identified nontuberculous mycobacterium capable of causing infections in both healthy and immunocompromised individuals. Only a few cases of M. septicum infections have been reported, which makes recognizing corresponding clinical disease more challenging for clinicians. Antimicrobial susceptibility profiles for this organism are not well described, and corresponding optimal therapeutic regimens have not been established. We report a tertiary care center's experience with M. septicum from 2014 to 2020. Twelve adult patients with positive cultures for M. septicum were identified. Most cases were identified from sputum samples of individuals with underlying lung disease. Most cases involving M. septicum isolation in culture were not felt to be clinically significant. Two cases were considered possible infections, while only one case was considered a definite infection that required antimicrobial treatment. All M. septicum isolates were susceptible in vitro to amikacin, ciprofloxacin, imipenem, linezolid, moxifloxacin, and trimethoprim-sulfamethoxazole. Isolates were universally resistant to clarithromycin and doxycycline. The isolation of M. septicum in culture is uncommon and requires clinical correlation to determine its clinical relevance and need for treatment. Susceptibility testing should be performed to guide therapy.


Loss of N-linked glycosylation reduces urea transporter UT-A1 response to vasopressin.
The vasopressin-regulated urea transporter (UT)-A1 is a transmembrane protein with two glycosylated forms of 97 and 117 kDa; both are derived from a single 88-kDa core protein. However, the precise molecular sites and the function for UT-A1 N-glycosylation are not known. In this study, we compared Madin-Darby canine kidney cells stably expressing wild-type (WT) UT-A1 to Madin-Darby canine kidney cell lines stably expressing mutant UT-A1 lacking one (A1m1, A1m2) or both glycosylation sites (m1m2). Site-directed mutagenesis revealed that UT-A1 has two glycosylation sites at Asn-279 and -742. Urea flux is stimulated by 10 nM vasopressin (AVP) or 10 microM forskolin (FSK) in WT cells. In contrast, m1m2 cells have a delayed and significantly reduced maximal urea flux. A 15-min treatment with AVP and FSK significantly increased UT-A1 cell surface expression in WT but not in m1m2 cells, as measured by biotinylation. We confirmed this finding using immunostaining. Membrane fractionation of the plasma membrane, Golgi, and endoplasmic reticulum revealed that AVP or FSK treatment increases UT-A1 abundance in both Golgi and plasma membrane compartments in WT but not in m1m2 cells. Pulse-chase experiments showed that UT-A1 half-life is reduced in m1m2 cells compared with WT cells. Our results suggest that mutation of the N-linked glycosylation sites reduces urea flux by reducing UT-A1 half-life and decreasing its accumulation in the apical plasma membrane. In vivo, inner medullary collecting duct cells may regulate urea uptake by altering UT-A1 glycosylation in response to AVP stimulation.


Maternal Smoking and Psychosocial Functioning: Impact on Subsequent Breastfeeding Practices.
Introduction: Breastfeeding is a protective factor for women and children. Women who smoke cigarettes during pregnancy are less likely to initiate or persist in breastfeeding. However, less is known about why this is the case. Materials and Methods: The present study (n = 247) prospectively examined maternal/child factors that influence breastfeeding in a low-income, racially diverse at-risk sample of smoking and nonsmoking women. Pregnant women were recruited at their first prenatal appointment in an urban hospital and followed through 24-month postnatally. Women reported on the average number of cigarettes smoked/day during pregnancy, psychopathology, breastfeeding behavior, and infant reactivity. Results: Although a greater number of cigarettes smoked/day during pregnancy was associated with a lower likelihood of initiating or persisting in breastfeeding, maternal age, education, and infant reactivity offered predictive utility above and beyond maternal smoking. Conclusion: Smokers were less likely to initiate breastfeeding and breastfed for shorter duration than demographically similar nonsmokers; however, one of the mechanisms for reduced breastfeeding may be the psychosocial factors of younger age and lower education. Further, infant reactivity was also found to reduce the likelihood of initiating and persisting with breastfeeding.


Trends in bacterial and fungal keratitis in South India, 2002-2012.
To assess the trends in microbiological organisms identified from corneal scrapings from patients with infectious keratitis at a tertiary care medical centre in South India. We reviewed the records of the microbiology laboratory at Aravind Eye Hospital in Madurai, India, from 2002 until 2012. We identified the microbiological causes of all corneal ulcers from the culture and smear results, and assessed for trends in bacterial and fungal keratitis over time. Of 23 897 corneal patients with ulcer with a corneal smear from 2002 to 2012 a fungal organism was identified in 34.3%, a bacterial organism in 24.7% and no organism in 38.3%. During this period, the annual number of keratitis cases due to bacteria decreased from 677 to 412, and the annual number due to fungus increased from 609 to 863. In analyses accounting for the total number of outpatients seen each year, the decline in number of smears positive for bacteria was statistically significant (p<0.001) but the increase in the number positive for fungus was not (p=0.73). The relative frequency of individual bacterial or fungal organisms remained relatively stable over this time. At a tertiary eye care centre in South India, there has been a reduction in the numbers of smear-positive bacterial keratitis over the past 11 years. This decline likely reflects economic development in India and increased access to antibiotics.


Frequent activation of c-kis as a transforming gene in fibrosarcomas induced by methylcholanthrene.
The DNA's from two of four methylcholanthrene-induced mouse fibrosarcomas contained transforming genes that were identical in their pattern of restriction endonuclease resistance to inactivation of biologic activity. This transforming gene was identified as the activated homolog of the Kirsten murine sarcoma virus onc gene, v-kis. The finding that a defined carcinogen reproducibly leads to activation of kis as a transforming gene should be of value in elucidating the role of oncogenes in the neoplastic process.


Herpes simplex-mediated gene transfer of nerve growth factor protects against peripheral neuropathy in streptozotocin-induced diabetes in the mouse.
Peripheral neuropathy is a common and debilitating complication of diabetes. In animal models, neurotrophic factors can prevent progression of the neuropathy, but adverse effects prevent systemic administration in adequate doses to treat human disease. We examined whether gene transfer with replication-defective genomic herpes simplex virus (HSV) vectors modified to express nerve growth factor (NGF) could be used to prevent progression of neuropathy in mice. Diabetes induced by streptozotocin (STZ) resulted in a sensory neuropathy manifest by a decrease in the foot sensory nerve amplitude (FSA; control = 20 +/- 0.1 microV, treated = 14 +/- 0.1 microV). Transduction of dorsal root ganglia in vivo with an HSV-based vector expressing NGF under the control of the human cytomegalovirus immediate early promoter (vector SHN) or the HSV latency active promoter 2 (vector SLN) by footpad inoculation 2 weeks after STZ administration protected against the decrease in FSA (22 +/- 1.4 microV and 21 +/- 1.7 microV, respectively) measured 4 weeks later. Injection of SHN into inguinal adipose tissue 2 weeks after onset of diabetes also prevented the decrease in FSA (20 +/- 3.3 microV). These results suggest that gene transfer with an NGF-producing herpes-based vector may prove useful in the treatment of diabetic neuropathy.


Potential Use of Nanomedicine for Drug Delivery Across the Blood-Brain Barrier in Healthy and Diseased Brain.
The research of efficacious non-invasive therapies for the treatment of brain diseases represents a huge challenge, as people affected by disorders of the central nervous system (CNS) will significantly increase. Moreover, the blood-brain barrier is a key factor in hampering a number of effective drugs to reach the CNS. This review is therefore focusing on possible interventions of nanomedicine-based approaches in selected diseases affecting the CNS. A wide overview of the most outstanding results on preclinical evaluations of the potential of nanomedicine in brain diseases (i.e. brain tumor, Alzheimer, Parkinson, epilepsy and others) is given, with highlights on the data with relevant interest and real possibility in translation from bench-to-bedside. Moreover, a critical evaluation on the rationale in planning nanosystems to target specific brain pathologies is described, opening the path to a more structured and pathology-tailored design of nanocarriers.


Assessing executive functions in traumatic brain injury: an exploratory study of the Executive Interview.
Executive dysfunction is a frequent sequela of traumatic brain injury. Two correlational studies using samples of inpatients and outpatients diagnosed with traumatic brain injury were undertaken to evaluate the validity of a bedside screening test of executive functioning, the 1992 Executive Interview EXIT 25 by Royall, Mahurin, and Gray. In the first study of 23 inpatients receiving rehabilitation, the EXIT 25 was strongly related to both the Mini-Mental State Examination and the Modified Mini-Mental State Examination. In the second study of 20 outpatients, the EXIT 25 correlated with other executive function measures and with ratings of functional disability at discharge, although a ceiling effect raised questions about its utility for patients with mild to moderate disability. Further study of the EXIT 25 is justified with a larger sample of inpatients, although caution should be exercised when using the EXIT 25 in mildly impaired outpatients.


Structural signature of the MYPT1-PP1 interaction.
Muscle relaxation is triggered by the dephosphorylation of Ser19 in the myosin regulatory light chain. This reaction is catalyzed by the holoenzyme myosin phosphatase (MP), which includes the catalytic subunit protein phosphatase 1 (PP1) and the regulatory targeting subunit (MYPT). MYPT1 (myosin phosphatase targeting subunit 1) is responsible for both targeting the holoenzyme to subcellular compartments in the muscle and directing PP1 specificity toward myosin. To understand the molecular events leading to the MYPT1-PP1 holoenzyme formation, we used NMR spectroscopy to determine the structural and dynamic characteristics of unbound MYPT1. This allowed the conformations of MYPT1 in the free, unbound state to be directly compared to the PP1-bound state. Our results show that MYPT1(1-98) behaves like a two-domain protein in solution. The first 40 residues of MYPT1(1-98), the disordered region, are intrinsically disordered and highly dynamic, whereas residues 41-98, the folded ankyrin-repeat region, are well-structured and rigid. Furthermore, the integrated use of NMR and biophysical data enabled us to calculate an ensemble model for MYPT1(1-98). The most prominent structural feature of the MYPT1(1-98) ensemble is a 25% populated transient α-helix in the disordered region of MYPT1(1-98). This α-helix becomes fully populated when bound to PP1 and, as we show, likely plays a central role in the formation of the MYPT1-PP1 holoenzyme complex. Finally, this combined analysis shows that the structural and dynamic behaviors exhibited by MYPT1 for PP1 are distinct from those of any other previously analyzed PP1 regulatory protein. Collectively, these data enable us to present a new model of the molecular events that drive MYPT1-PP1 holoenzyme formation and demonstrate that there are structural differences in unbound PP1 regulators that have not been previously observed. Thus, this work adds significant insights to the currently limited data for molecular structures and dynamics of PP1 regulators.


The Cytomegalovirus-Specific IL-21 ELISpot Correlates with Allograft Function of Kidney Transplant Recipients.
In kidney transplant recipients, the cytomegalovirus (CMV) is frequently causing infection/reactivation and can trigger allograft rejection. To assess the risk of reactivation, the cellular immune response against CMV is increasingly assessed by cellular in vitro methods, such as the interferon (IFN)-γ ELISpot. In the current study we compared the IFN-γ ELISpot with our newly established CMV-specific ELISpot assays determining IL-17A, IL-21, IL-22, granzyme B, and perforin and correlated the results with flow cytometric data and clinical parameters. In 77 kidney transplant recipients, the highest frequency was observed for CMV pp65-specific cells secreting IFN-γ, followed by cells secreting IL-21 (62.9 and 23.2 Δ spot forming cells/10⁵ cells). We observed a positive correlation between the percentage of CMV-specific CD3+ CD4+ CD154+ cells and results of the CMV-specific IL-21 ELISpot (p = 0.002). Results of the CMV pp65-specific IL-21 ELISpot correlated negatively with kidney function (estimated glomerular filtration rate, p = 0.006) and were significantly higher in women (p = 0.005). IL-21, a cytokine involved in aging that is secreted by activated CD4+ T cells, may also impact on allograft function. Thus, the CMV-specific IL-21 ELISpot could become a new tool to assess if CMV seropositivity represents a hazard for the graft.


Local apoptosis promotes collagen production by monocyte-derived cells in transforming growth factor β1-induced lung fibrosis.
Collagen-containing leukocytes (CD45+Col-I+) accumulate in diseased and fibrotic tissues. However, the precise identity of these cells and whether injury is required for their recruitment remain unknown. Using a murine model of pulmonary fibrosis in which an inducible, bioactive form of the human transforming growth factor (TGF)-β1 gene is targeted to the lung, we characterized the cell surface phenotype of collagen-containing CD45+ cells in the lung and tested the hypothesis that apoptotic cell death responses are essential to the accumulation of CD45+Col-I+ cells. Our studies demonstrate that CD45+Col-I+ cells appearing in the TGF-β1-exposed murine lung express markers of the monocyte lineage. Inhibition of apoptosis via pharmacological caspase blockade led to a significant reduction in CD45+Col-I+ cells, which appear to accumulate independently of alternatively activated macrophages. There are also increased levels of apoptosis and greater numbers of CD45+Col-I+ in the lung tissue of patients with two distinct forms of fibrotic lung disease, idiopathic pulmonary fibrosis and connective tissue disease-related interstitial lung disease, when compared to lung from healthy normal controls. These findings are accompanied by an increase in collagen production in cultured monocytes obtained from subjects with fibrotic lung disease. Treatment of these cultured cells with the caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD/fmk) reduces both apoptosis and collagen production in all subjects. Interventions that prevent collagen production by monocytes via modulation of caspase activation and of apoptosis may be ameliorative in monocyte-associated, TGF-β1-driven processes such as pulmonary fibrosis.


Postintervention serum TSH levels may be useful to differentiate patients who should undergo levothyroxine suppressive therapy after thyroid surgery for multinodular goiter in a region with moderate iodine deficiency.
Recent studies have raised doubts about the efficacy of the postoperative use of levothyroxine (LT4) suppressive doses in patients who underwent thyroid surgery for multinodular goiter. The purpose of this retrospective study was to examine the efficacy of different doses of LT4 in preventing postsurgical recurrences of simple multinodular goiter and to identify a marker that could be useful in discriminating patients with a higher risk of developing recurrence. Two hundred thirty-two patients (57 male, 175 female) operated for nontoxic multinodular goiter were divided into two groups: (I) patients with normal postsurgery thyrotropin (TSH) levels (0.25 to 4.5 mU/L) and (II) patients with elevated postsurgery TSH levels (>4.5 mU/L). All patients were subjected to replacement (1.3 microg LT4/kg/day) or suppressive (1.7 microg LT4/kg/day) doses of LT4, and they were followed for a median period of 6 years (range 2 to 12). No statistical difference was found for sex, age, and postsurgery serum TSH between patients submitted to suppressive and replacement therapy. The ultrasound (US) detection of new postsurgery nodules of at least 0.5 cm maximum diameter was considered a recurrence of disease and was found in 10% of the cases studied. Patients with normal postsurgery serum TSH showed a high recurrence rate (30.4%) when submitted to lower daily doses of LT4. In patients with elevated postsurgery serum TSH, the rate of nodular goiter recurrence did not vary with different types of LT4 therapy. In conclusion, our results suggest that the postsurgical serum TSH is useful for prediction of nodular goiter recurrence, as it reflects the amount of residual functioning thyroid tissue in the cervical area. It may also be indicative of patients who might benefit from LT4 suppressive therapy.


Soybean genetics, genomics, and breeding for improving nutritional value and reducing antinutritional traits in food and feed.
Soybean [Glycine max (L.) Merr.] is a globally important crop due to its valuable seed composition, versatile feed, food, and industrial end-uses, and consistent genetic gain. Successful genetic gain in soybean has led to widespread adaptation and increased value for producers, processors, and consumers. Specific focus on the nutritional quality of soybean seed composition for food and feed has further elucidated genetic knowledge and bolstered breeding progress. Seed components are historical and current targets for soybean breeders seeking to improve nutritional quality of soybean. This article reviews genetic and genomic foundations for improvement of nutritionally important traits, such as protein and amino acids, oil and fatty acids, carbohydrates, and specific food-grade considerations; discusses the application of advanced breeding technology such as CRISPR/Cas9 in creating seed composition variations; and provides future directions and breeding recommendations regarding soybean seed composition traits.


Analysis of fatigue and tremor during sustained maximal grip contractions using Hilbert-Huang Transformation.
The objective of this study was to evaluate muscle fatigue and tremor during a Sustained Maximal Grip Contraction (SMGC) using the Hilbert-Huang Transformation (HHT). Thirty-nine healthy subjects volunteered for the study and performed a 25-s SMGC. Fatigue parameters such as the relative force output (RFO) were calculated from the residual of SMGC after applying Empirical Mode Decomposition (EMD). Using the energy spectrum of the Intrinsic Mode Functions (IMF) obtained using HHT, isometric force tremor was identified from the 4 to 12 Hz region in IMF3 and IMF4. Data were analysed for five consecutive 5-s epochs to identify changes in fatigue and tremor over time. The HHT method was able to identify a greater resistance to fatigue in women compared to men (p≤0.05) and in non-dominant hands compared to dominant hands (p≤0.05). Consistent with the results for fatigue, women had less tremor than men (p≤0.05), while non-dominant hands trembled less than did dominant hands (p≤0.05). Higher levels of tremor were observed for non-fatigue-resistant subjects for both 10-15 s and 15-20 s epochs (p≤0.05). The HHT is an appropriate method to identify both fatigue and tremor during SMGC. It would be of interest to apply this method to the study the elderly or patients with neuromuscular disorders.


"Causes of death in asthma, COPD and non-respiratory hospitalized patients: a multicentric study".
There is limited information on the causes of death in asthma patients.To determine the causes of death in hospitalized asthmatic patients and to compare with those observed in COPD patients and non-respiratory individuals, with a particular interest in associations with previous cardiovascular disease. Retrospective case-control study which analyzed the deaths of all hospitalized patients admitted for any reason during January, April, July and October of 2008 in 13 Spanish centers. Medical records of deceased patients were reviewed, and demographic and clinical data were collected. A total of 2,826 deaths (mean age 75 years, 56% men) were included in the analysis, of which 82 (2.9%) were of patients with asthma and 283 (10%) with COPD.The most common causes of death in asthma patients were cardiovascular diseases (29.3%), malignancies (20.7%) and infections (14.6%); in COPD patients they were malignancies (26.5%), acute respiratory failure (25.8%) and cardiovascular diseases (21.6%). Asthma, compared to COPD patients, died significantly less frequently from acute respiratory failure and lung cancer. A multivariate logistic regression analysis failed to associate asthma with cardiovascular deaths. Cardiovascular disease is the most frequent cause of death among hospitalized asthma patients. The specific causes of death differ between asthma and COPD patients.


Alpha-defensins inhibit ERK/STAT3 signaling during monocyte-macrophage differentiation and impede macrophage function.
Alpha-1-antitrypsin deficiency (AATD) is a genetic disorder associated with a 5-tenfold decrease in lung levels of alpha-1-antitrypsin (AAT) and an increased risk for obstructive lung disease. α-defensins are cationic broad-spectrum cytotoxic and pro-inflammatory peptides found in the azurophilic granules of neutrophils. The concentration of α-defensins is less than 30 nM in the bronchoalveolar lavage fluid of healthy controls but is up to 6 μM in AATD individuals with significant lung function impairment. Alveolar macrophages are generally classified into pro-inflammatory (M1) or anti-inflammatory (M2) subsets that play distinct roles in the initiation and resolution of inflammation. Therefore, monocyte-macrophage differentiation should be tightly controlled to maintain lung integrity. In this study, we determined the effect of α-defensins on monocyte-macrophage differentiation and identified the molecular mechanism of this effect. The results of this study demonstrate that 2.5 μM of α-defensins inhibit the phosphorylation of ERK1/2 and STAT3 and suppress the expression of M2 macrophage markers, CD163 and CD206. In addition, a scratch assay shows that the high concentration of α-defensins inhibits cell movement by ~ 50%, and the phagocytosis assay using flow cytometry shows that α-defensins significantly reduce the bacterial phagocytosis rate of monocyte-derived macrophages (MDMs). To examine whether exogenous AAT is able to alleviate the inhibitory effect of α-defensins on macrophage function, we incubated MDMs with AAT prior to α-defensin treatment and demonstrate that AAT improves the migratory ability and phagocytic ability of MDMs compared with MDMs incubated only with α-defensins. Taken together, this study suggests that a high concentration of α-defensins inhibits the activation of ERK/STAT3 signaling, negatively regulates the expression of M2 macrophage markers, and impairs innate immune function of macrophages.


Control of compound leaf patterning by MULTI-PINNATE LEAF1 (MPL1) in chickpea.
Plant lateral organs are often elaborated through repetitive formation of developmental units, which progress robustly in predetermined patterns along their axes. Leaflets in compound leaves provide an example of such units that are generated sequentially along the longitudinal axis, in species-specific patterns. In this context, we explored the molecular mechanisms underlying an acropetal mode of leaflet initiation in chickpea pinnate compound leaf patterning. By analyzing naturally occurring mutants multi-pinnate leaf1 (mpl1) that develop higher-ordered pinnate leaves with more than forty leaflets, we show that MPL1 encoding a C2H2-zinc finger protein sculpts a morphogenetic gradient along the proximodistal axis of the early leaf primordium, thereby conferring the acropetal leaflet formation. This is achieved by defining the spatiotemporal expression pattern of CaLEAFY, a key regulator of leaflet initiation, and also perhaps by modulating the auxin signaling pathway. Our work provides novel molecular insights into the sequential progression of leaflet formation.


Saturation of mismatch repair in the mutD5 mutator strain of Escherichia coli.
The mutD (dnaQ) gene of Escherichia coli codes for the proofreading activity of DNA polymerase III. The very strong mutator phenotype of mutD5 strains seems to indicate that their postreplicational mismatch repair activity is also impaired. We show that the mismatch repair system of mutD5 strains is functional but saturated, presumably by the excess of DNA replication errors, since it is recovered by inhibiting chromosomal DNA replication. This recovery depends on de novo protein synthesis.


What's love got to do with it? Social functioning, perceived health, and daily happiness in married octogenarians.
This study examined day-to-day links between perceived health and happiness and between time spent with others and happiness in 47 older adult couples over an 8-day period. Marital satisfaction and time spent with others were explored as potential moderators of links between health and happiness. For both men and women, hierarchical linear modeling revealed daily links between more time spent with others and greater happiness. Daily links between time spent with one's partner and happiness were strongly moderated by marital satisfaction. For both men and women, marital satisfaction buffered day-to-day links between poorer perceived health and a decline in happiness, but time spent with others did not. This study provides support for the role of marital satisfaction in protecting older adults' happiness from daily fluctuations in perceived physical health and for the influence of social connections in promoting happiness in the lives of older adults. (PsycINFO Database Record (c) 2010 APA, all rights reserved).


Variability of the cranial and dental phenotype in Williams syndrome.
Williams syndrome (WS) is a rare congenital disorder involving the cardiovascular system, connective tissue, and the central nervous system, resulting in mild to moderate mental retardation, a specific cognitive profile, unique personality characteristics, distinctive facial features, and cardiovascular disease. The majority of individuals with a clinical diagnosis of WS have a contiguous gene deletion at chromosome 7 (7q11.23). Physical features include characteristic facial features with full prominent cheeks, wide mouth, long philtrum, small nose with depressed nasal bridge, heavy orbital ridges, medial eyebrow flare, dental abnormalities, hoarse voice, growth retardation, and cardiovascular abnormalities (most commonly supravalvular aortic stenosis and/or peripheral pulmonary artery stenosis). The cognitive profile is distinctive, consisting of strengths in auditory memory, language, and face-processing, but extreme weakness in visuospatial, numerical and problem-solving abilities. Neurological studies have identified a significantly decreased brain volume in adult individuals with WS with relatively normal development of the limbic, frontal and cerebellar structures. The aims were to analyse the neurocranium, the craniofacial region, and the dentition in a well defined Norwegian group of individuals with WS. In order to accomplish this, normative cephalometric standards for the neurocranium, including the cranial base and the sella turcica, were established for Norwegian males and females from 6 to 21 years of age, using lateral radiographic cephalograms from the Oslo University Craniofacial Growth Archive. The study material comprised radiographic lateral cephalograms, orthopantomograms and dental casts from 62 individuals with WS ranging from 4 to 44 years. The lateral cephalograms, orthopantomograms and dental casts were analysed using standard methods reported in the literature. Neurocranium: The results from the cephalometric analyses showed that the size and shape of the neurocranium in WS differed from normal controls. A flattening of the superior aspect of the parietal bone was found and the posterior prominence of the occipital bone was larger. These findings were in concordance with published neuroanatomical/neuropathological studies on WS and fit well with the reported anatomical aberrations in the WS brain, e.g. a smaller brain volume where some parts are of relatively normal size while others are decreased. The anterior and posterior cranial base was shorter in WS, but with a normal cranial base angle. The thickness of the calvarian bones, especially the frontal and occipital bones, was greater in WS than in normal controls. Sella turcica: The size of the sella turcica was somewhat smaller in WS compared with normal controls, though not significant. Aberrant types of shape of the sella turcica in WS are demonstrated. The correlation between prenatal malformation in the pituitary gland/sella turcica and the postnatal morphology of the sella turcica is discussed. The craniofacial skeleton: Two areas, in addition to the cranial base, have been identified to contribute to the characteristic facial appearance in WS; the anterior inclination of the maxilla, and the shape of the mandible. The severely deficient bony chin in combination with the high mandibular plane angle can explain earlier clinical descriptions of a retrusive mandible in WS. Frequent mouth breathing during childhood and the smaller cranial base may be associated with the characteristic craniofacial pattern. Dentition: In individuals with WS over 10 years of age 40.5 per cent had agenesis of one or more permanent teeth and 11.9 percent had agenesis of 6 teeth or more. The missing permanent teeth in the maxilla most commonly included 2nd premolars, 1st premolars, and lateral incisors. In the mandible most commonly 2nd premolars, 1st premolars, and central incisors were absent. The tooth crowns of permanent teeth were smaller, both in the mesio-distal and labio-lingual dimension. The permanent tooth crowns have several characteristic deviations from normal shape; especially incisors in both jaws were tapered or screwdriver-shaped. An evaluation of taurodontism on permanent molars was performed. However, most of the molars rated as being taurodontic had short total tooth lengths and could thus be rated taurodontic without meeting the classical definition. These studies have shown aberrations in the neurocranium including the sella turcica in WS not previously reported. In the craniofacial skeleton specific traits have been identified to contribute to the characteristic facial appearance in WS. Dental aberrations in the permanent dentition including agenesis, smaller size, and aberrant shape of teeth, are common in WS.


Screening for the prevention and early detection of cervical cancer: protocol for systematic reviews to inform Canadian recommendations.
To inform recommendations by the Canadian Task Force on Preventive Health Care on screening in primary care for the prevention and early detection of cervical cancer by systematically reviewing evidence of (a) effectiveness; (b) test accuracy; (c) individuals' values and preferences; and (d) strategies aimed at improving screening rates. De novo reviews will be conducted to evaluate effectiveness and to assess values and preferences. For test accuracy and strategies to improve screening rates, we will integrate studies from existing systematic reviews with search updates to the present. Two Cochrane reviews will provide evidence of adverse pregnancy outcomes from the conservative management of cervical intraepithelial neoplasia. We will search Medline, Embase, and Cochrane Central (except for individuals' values and preferences, where Medline, Scopus, and EconLit will be searched) via peer-reviewed search strategies and the reference lists of included studies and reviews. We will search ClinicalTrials.gov and the World Health Organization International Clinical Trials Registry Platform for ongoing trials. Two reviewers will screen potentially eligible studies and agree on those to include. Data will be extracted by one reviewer with verification by another. Two reviewers will independently assess risk of bias and reach consensus. Where possible and suitable, we will pool studies via meta-analysis. We will compare accuracy data per outcome and per comparison using the Rutter and Gatsonis hierarchical summary receiver operating characteristic model and report relative sensitivities and specificities. Findings on values and preferences will be synthesized using a narrative synthesis approach and thematic analysis, depending on study designs. Two reviewers will appraise the certainty of evidence for all outcomes using GRADE (Grading of Recommendations Assessment, Development and Evaluation) and come to consensus. The publication of guidance on screening in primary care for the prevention and early detection of cervical cancer by the Task Force in 2013 focused on cytology. Since 2013, new studies using human papillomavirus tests for cervical screening have been published that will improve our understanding of screening in primary care settings. This review will inform updated recommendations based on currently available studies and address key evidence gaps noted in our previous review.


Leadership skills for the multi-tiered nursing team.
In a challenging global healthcare climate, new workforce models are required to address the ongoing shortfall in the number of nurses. One way of addressing the workforce crisis in the NHS has been to introduce the roles of nursing associate and assistant practitioner, which are designed to support registered healthcare professionals in their work and 'bridge the gap' between registered professionals and support workers. This article examines the leadership skills required of nurses when seeking to embed these roles in practice, and the leadership skills assistant practitioners and nursing associates themselves need to take their place in the team. The author also discusses the need for nurses to adopt a relational leadership style, hone their delegation skills and foster a climate of psychological safety for their assistant practitioner and nursing associate colleagues.


A Privacy-Preserving Infrastructure for Analyzing Personal Health Data in a Vertically Partitioned Scenario.
It is widely anticipated that the use and analysis of health-related big data will enable further understanding and improvements in human health and wellbeing. Here, we propose an innovative infrastructure, which supports secure and privacy-preserving analysis of personal health data from multiple providers with different governance policies. Our objective is to use this infrastructure to explore the relation between Type 2 Diabetes Mellitus status and healthcare costs. Our approach involves the use of distributed machine learning to analyze vertically partitioned data from the Maastricht Study, a prospective population-based cohort study, and data from the official statistics agency of the Netherlands, Statistics Netherlands (Centraal Bureau voor de Statistiek; CBS). This project seeks an optimal solution accounting for scientific, technical, and ethical/legal challenges. We describe these challenges, our progress towards addressing them in a practical use case, and a simulation experiment.


Effects of calcium, lanthanum, and bicarbonate ion on epinephrine modification of insulin release in vitro.
The role of calcium flux in mediating epinephrine modification of insulin release was investigated by using lanthanum, an inhibitor of calcium flux, in the in vitro perifusion system. Lanthanum inhibits insulin secretion stimulated by glucose and by acetylcholine to basal levels. Epinephrine and lanthanum have additive effects in inhibiting insulin secretion to glucose stimulation. The effect of epinephrine prestimulation on insulin secretion to subsequent glucose challenge varies markedly, depending on the presence or absence of bicarbonate ion: epinephrine priming is reversed in the absence of bicarbonate, and effect possibly related to reduced calcium uptake. In the presence of bicarbonate, lanthanum blocks the priming effect of epinephrine on insulin secretion. The data further support the postulated role of calcium in adrenergic effects on beta cell function.


[Dalbavancin and its use in the treatment of methicillin-resistant Staphylococcus aureus - induced upper limb phlegmon].
The article discusses dalbavancin, a relatively new lipoglycopeptide antibiotic brought to market. It briefly describes the spectrum and mechanism of its antibacterial effect and dosing regimens that can be used in the treatment of skin and soft tissue infections. The authors present a case of a patient with shoulder phlegmon caused by MRSA who was successfully treated with this antibiotic.


[Disorders of hemostasis in peptic ulcer of the stomach and duodenum].
Eighty-seven patients with complicated patterns of peptic ulcer were examined for different components of the hemostatic system including gastric and duodenal fibrinolysis. It was established that the penetrating ulcer gives rise to hypocoagulemia and accelerates fibrinolysis. The perforating and hemorrhagic ulcers are associated with hypercoagulation, which is a manifestation of the stress response of the body. It is shown that gastric juice and the duodenal content have a hypocoagulation (heparinoid) action on blood and plasma coagulation. Peptic ulcer is marked by high gastric and duodenal fibrinolysis and proteolysis both in the gastric and duodenal mucosa and in the free gastric and duodenal contents as well. It is postulated that all the factors under consideration, which are unfavourable for local hemostasis, may be involved in the pathogenesis of gastroduodenal hemorrhages. The author holds that hemostatics, which suppress fibrinolysis and the acid-peptic factor and stimulate blood coagulation and clot formation in the zone of hemorrhage, may be used locally.


Oxygenation of the endocannabinoid, 2-arachidonylglycerol, to glyceryl prostaglandins by cyclooxygenase-2.
Cyclooxygenases (COX) play an important role in lipid signaling by oxygenating arachidonic acid to endoperoxide precursors of prostaglandins and thromboxane. Two cyclooxygenases exist which differ in tissue distribution and regulation but otherwise carry out identical chemical functions. The neutral arachidonate derivative, 2-arachidonylglycerol (2-AG), is one of two described endocannabinoids and appears to be a ligand for both the central (CB1) and peripheral (CB2) cannabinoid receptors. Here we report that 2-AG is a substrate for COX-2 and that it is metabolized as effectively as arachidonic acid. COX-2-mediated 2-AG oxygenation provides the novel lipid, prostaglandin H(2) glycerol ester (PGH(2)-G), in vitro and in cultured macrophages. PGH(2)-G produced by macrophages is a substrate for cellular PGD synthase, affording PGD(2)-G. Pharmacological studies reveal that macrophage production of PGD(2)-G from endogenous sources of 2-AG is calcium-dependent and mediated by diacylglycerol lipase and COX-2. These results identify a distinct function for COX-2 in endocannabinoid metabolism and in the generation of a new family of prostaglandins derived from diacylglycerol and 2-AG.


Residues involved in the pore-forming activity of the Clostridium perfringens iota toxin.
Clostridium perfringens iota toxin is a binary toxin that is organized into enzyme (Ia) and binding (Ib) components. Ib forms channels in lipid bilayers and mediates the transport of Ia into the target cells. Here we show that Ib residues 334-359 contain a conserved pattern of alternating hydrophobic and hydrophilic residues forming two amphipathic β-strands involved in membrane insertion and channel formation. This stretch of amino acids shows remarkable structural and functional analogies with the β-pore-forming domain of C. perfringens epsilon toxin. Several mutations within the two amphipathic β-strands affected pore formation, single-channel conductance and ion selectivity (S339E-S341E, Q345H N346E) confirming their involvement in channel formation. F454 of Ib corresponds to the Φ-clamp F427 of anthrax protective antigen and F428 of C2II binary toxins. The mutation F454A resulted in a loss of cytotoxicity and strong increase in single-channel conductance (500 pS as compared with 85 pS in 1 M KCl) with a slight decrease in cation selectivity, indicating that the Φ-clamp is highly conserved and crucial for binary toxin activity. In contrast, the mutants Q367D, N430D, L443E had no or only minor effects on Ib properties, while T360I, T360A and T360W caused a dramatic effect on ion selectivity and single-channel conductance, indicating gross disturbance of the oligomer structure. This suggests that, at least in the iota toxin family, T360 has a structural role in the pore organization. Moreover, introduction of charged residues within the channel (S339E-S341E) or in the vestibule (Q367D, N430D and L443E) had virtually no effect on chloroquine or Ia binding, whereas F454A, T360I, T360A and T360W strongly decreased the chloroquine and Ia affinity to Ib. These results support that distinct residues within the vestibule interact with chloroquine and Ia or are responsible for channel structure, while the channel lining amino acids play a less important role.


Cannabis, Decision-Making, and Online Assistance Seeking.
Maladaptive decision-making strategies could contribute to cannabis-related problems, as some individuals may neither select safe patterns of cannabis use, nor seek treatment. To explore decision-making styles and their relationship to cannabis-related harm, 153 respondents completed the Cannabis Use Disorders Identification Test-Revised (CUDIT-R), the Melbourne Decision Making Questionnaire (MDMQ), and answered questions about their willingness to seek online: (1) further information or (2) treatment for cannabis-related issues. Multiple regression considered relationships between problematic cannabis use, decision-making style, and cannabis use within the past month. Subscales of the CUDIT-R revealed that: (1) hazardous use was associated with higher hypervigilance and higher decisional self-esteem; (2) dependence symptoms were associated with lower vigilance and higher procrastination; and (3) harmful use was associated with higher procrastination. People with symptoms of CUD were less likely to seek further help or support online if prone to procrastination or buckpasssing. Decisional style influenced cannabis use and symptoms. Those people with cannabis-related problems that did not seek online assistance were defensively avoidant. Procrastination is a feature of problematic cannabis use. Online offers of assistance may be ignored by defensively avoidant CUD clients. (Am J Addict 2019;00:1-7).


Novel proteomic biomarker panel for prediction of aggressive metastatic hepatocellular carcinoma relapse in surgically resectable patients.
The natural course of early HCC is unknown, and its progression to intermediate and advanced HCC can be diverse. Some early stage HCC patients enjoy prolonged disease-free survival, whereas others suffer aggressive relapse to stage IV metastatic cancer within a year. Comparative proteomics of HCC tumor tissues was carried out using 2D-DIGE and MALDI-TOF/TOF MS to identify proteins that can distinguish these two groups of stage I HCC patients. Twelve out of 148 differentially regulated protein spots were found to differ by approximately 2-fold for the relapse versus nonrelapse patient tissues. Four proteins, namely, heat shock 70 kDa protein 1, argininosuccinate synthase, isoform 2 of UTP-glucose-1-phosphate uridylyltransferase, and transketolase, were shown to have the potential to differentiate metastatic relapse (MR) from nonrelapse (NR) HCC patients after validation by western blotting and immunohistochemical assays. Subsequent TMA analysis revealed a three marker panel of HSP70, ASS1, and UGP2 to be statistically significant in stratifying the two groups of HCC patients. This combination panel achieved high levels of sensitivity and specificity, which has potential for clinical use in identifying HCC tumors prone to MR. This stratification will allow development of clinical management, including close follow-up and possibly treatment options, in the near future.


Resolution of trauma-related guilt following treatment of PTSD in female rape victims: a result of cognitive processing therapy targeting comorbid depression?
Although Resick et al. [Resick, P.A., Nishith, P., Weaver, T.L., Astin, M.C., Feuer, C.A., 2002. A comparison of cognitive-processing therapy with prolonged exposure and a waiting condition for the treatment of chronic posttraumatic stress disorder in female rape victims. J. Consult. Clin. Psychol. 70, 867-879.] reported comparable results for treating rape-related posttraumatic stress disorder (PTSD) using either cognitive-processing therapy (CPT) or prolonged exposure (PE), there was some suggestion that CPT resulted in better outcomes than PE for certain aspects of trauma-related guilt. The present study revisited these findings to examine whether this effect was a function of improvement in a subset of participants with both PTSD and major depressive disorder (MDD). Results indicated that CPT was just as effective in treating "pure" PTSD and PTSD with comorbid MDD in terms of guilt. Clinical significance testing underscored that CPT was more effective in reducing certain trauma-related guilt cognitions than PE. Findings cannot be generalized to men, and only one measure of guilt was used. The observed superiority of CPT over PE for treating certain guilt cognitions was not due to participant comorbidity. Further research is recommended to untangle the relationship between guilt, depression and differential response to treatment in PTSD following sexual assault trauma.


Critical transitions in Chinese dunes during the past 12,000 years.
Dune systems can have alternative stable states that coexist under certain environmental conditions: a vegetated, stabilized state and a bare active state. This behavior implies the possibility of abrupt transitions from one state to another in response to gradual environmental change. Here, we synthesize stratigraphic records covering 12,000 years of dynamics of this system at 144 localities across three dune fields in northern China. We find side-by-side coexistence of active and stabilized states, and occasional sharp shifts in time between those contrasting states. Those shifts occur asynchronously despite the fact that the entire landscape has been subject to the same gradual changes in monsoon rainfall and other conditions. At larger scale, the spatial heterogeneity in dune dynamics averages out to produce relatively smooth change. However, our results do show different paths of recovery and collapse of vegetation at system-wide scales, implying that hysteretic behavior occurs in spatially extended systems.


Serial analysis of gene expression in mice with lipopolysaccharide-induced acute lung injury.
To monitor the systemic gene expression profile in a murine model of lipopolysaccharide-induced acute lung injury. Acute lung injury was induced by intratracheal injection of lipopolysaccharide in 3 mice. Another 3 normal mice receiving same volume of normal saline were taken as the controls. The comprehensive gene expression profile was monitored by the recently modified long serial analysis of gene expression. A total of 24,670 tags representing 12,168 transcripts in the control mice and 26,378 tags representing 13,397 transcripts in the mice with lung injury were identified respectively. There were 11 transcripts increasing and 7 transcripts decreasing more than 10 folds in the lipopolysaccharide-treated mice. The most overexpressed genes in the mice with lung injury included serum amyloid A3, metallothionein 2, lipocalin 2, cyclin-dependent kinase inhibitor 1A, lactate dehydrogenase 1, melatonin receptor, S100 calcium-binding protein A9, natriuretic peptide precursor, etc. Mitogen activated protein kinase 3, serum albumin, complement component 1 inhibitor, and ATP synthase were underexpressed in the lung injury mice. Serial analysis of gene expression provides a molecular characteristic of acute lung injury.


[Medication: ethical and legal aspects for nursing].
This article treats of the duties and responsibilities of nursing staff concerning medication process. From a survey about the valid legislation, the authors made considerations about the legal implications to the nursing staff, especially related to the assistance quality detours involving the medication process, as well as the contradictions in law that restrict the information access about medication to these professionals.


Diabetes in pregnancy: rationale and guidelines for care.
The management of pregnancy in a woman with diabetes is directed toward insuring a normal outcome for the mother and her newborn baby. There is ample evidence from studies in both humans and animals that maintenance of a normal maternal metabolic environment, particularly blood glucose concentrations, can avoid or ameliorate the complications of pregnancy responsible for increased morbidity and mortality in the mother and her baby. It is essential that this metabolic normalcy be achieved preconceptionally if congenital malformations, the leading cause of death in IDMs, are to be avoided. Thus, preconceptional care, intensive regulation of maternal glucose metabolism and fetal surveillance throughout pregnancy are critical. The use of the multidisciplinary team to optimize care has led to significant reductions in maternal and perinatal morbidity/mortality in IDMs, and is thus essential to successful outcomes in pregnancies complicated by diabetes.


D88A mutant of cytochrome P450nor provides kinetic evidence for direct complex formation with electron donor NADH.
The haem-distal pocket of nitric oxide reductase cytochrome P450 contains many Arg and Lys residues that are clustered to form a putative access channel for NADH. Asp88 is the sole negatively charged amino acid in this positive charge cluster, and thus it would be interesting to know its functional role. Here we found the intriguing phenomenon that mutation at this site of P450nor (D88A or D88V) considerably decreased the overall nitric oxide reductase activity without blocking the reducing half reaction in which the ferric enzyme-NO complex is reduced with NADH to yield a specific intermediate (I). The results indicate that the catalytic turnover subsequent to the I formation was blocked by such mutation. This property of the mutants made it possible to perform kinetic analysis of the reduction step, which is impossible with the wild-type P450nor. These results are the first kinetic evidence for direct complex formation between P450nor and an electron donor (NADH or NADPH). The kinetic analysis also showed that the inhibition by chloride ions (Cl(-)) is competitive with respect to NAD(P)H, which highlights the importance of the binding site for Cl(-) (the anion hole) in the interaction with NAD(P)H. We also characterized another mutant (D393A) of P450nor. The results demonstrated that both Asp residues play important roles in the interaction with NADH, whereas the role of Asp88 is unique in that it must be essential for the release of NAD(+) rather than binding to NADH.


A hybrid deconvolution approach for estimation of in vivo non-displaceable binding for brain PET targets without a reference region.
Estimation of a PET tracer's non-displaceable distribution volume (VND) is required for quantification of specific binding to its target of interest. VND is generally assumed to be comparable brain-wide and is determined either from a reference region devoid of the target, often not available for many tracers and targets, or by imaging each subject before and after blocking the target with another molecule that has high affinity for the target, which is cumbersome and involves additional radiation exposure. Here we propose, and validate for the tracers [11C]DASB and [11C]CUMI-101, a new data-driven hybrid deconvolution approach (HYDECA) that determines VND at the individual level without requiring either a reference region or a blocking study. HYDECA requires the tracer metabolite-corrected concentration curve in blood plasma and uses a singular value decomposition to estimate the impulse response function across several brain regions from measured time activity curves. HYDECA decomposes each region's impulse response function into the sum of a parametric non-displaceable component, which is a function of VND, assumed common across regions, and a nonparametric specific component. These two components differentially contribute to each impulse response function. Different regions show different contributions of the two components, and HYDECA examines data across regions to find a suitable common VND. HYDECA implementation requires determination of two tuning parameters, and we propose two strategies for objectively selecting these parameters for a given tracer: using data from blocking studies, and realistic simulations of the tracer. Using available test-retest data, we compare HYDECA estimates of VND and binding potentials to those obtained based on VND estimated using a purported reference region. For [11C]DASB and [11C]CUMI-101, we find that regardless of the strategy used to optimize the tuning parameters, HYDECA provides considerably less biased estimates of VND than those obtained, as is commonly done, using a non-ideal reference region. HYDECA test-retest reproducibility is comparable to that obtained using a VND determined from a non-ideal reference region, when considering the binding potentials BPP and BPND. HYDECA can provide subject-specific estimates of VND without requiring a blocking study for tracers and targets for which a valid reference region does not exist.


Robotic Thymectomy: Learning Curve and Associated Perioperative Outcomes.
Recently, robotic-assisted thymectomy (RAT) has emerged as an alternative to either, an open transsternal approach or to a video-assisted thoracoscopic approach, for both thymic tumors and benign lesions. We have reviewed our early experience with RAT to assess the associated learning curve as well as the short-term perioperative outcomes. A prospectively collected database was reviewed for patients who underwent RAT for all causes in the period 2012-2016. Robotic thymectomy cases were stratified and compared according to the number of cases performed by each surgeon (≤15 versus >15 cases). A propensity score matching was done to compare perioperative outcomes in patients undergoing robotic and transsternal resection of thymomas. Seventy patients (47 females) with a median age of 52, underwent RAT. The median operative time was 102 min with 5 conversions to an open approach for local invasion (n = 3) or for complete pleural symphysis (n = 2). There were 2 rib fractures and 1 recurrent laryngeal nerve palsy. Median length of chest tube drainage and length of stay were 1 and 3 days, respectively. Operative time and estimated blood loss plateaued after surgeon's initial 15-20 cases, which may reflect the initial learning curve. A comparison between early and late robotic cases showed that with the growing experience, the operative time becomes shorter (94 versus 107 min, P = .018). Propensity score analysis between robotic and transsternal resection of thymoma (n = 22 in each group) showed no significant differences in operative time (P = .79), intraoperative complications (P = .99), or postoperative complications (P = .99). Robotic thymectomy is feasible and safe, and is associated with comparable perioperative outcomes to the traditional transsternal approach in patients undergoing thymomectomy. An initial learning curve of 15-20 robotic thymectomy cases may be required by the surgeons to adequately perform this relatively novel technique.


Extremely Long-Range Light-Driven Repulsion of Porous Microparticles.
The repulsive surface forces, such as electrostatic or steric, acting between particles explain why they remain well separated in aqueous electrolyte solutions and are responsible for the stability of colloidal dispersions. However, the effective range of these interactions is always well below hundreds of nanometers and typically can be controlled by advanced manipulations such as tuning the electrolyte concentration or modifying the particle surface or, in some more specific cases, via subjecting the suspension to an external electric or magnetic field. Here we employ solutions with small additives of a photosensitive ionic surfactant to investigate if a repulsive interaction of microsized particles sedimented at the solid surface can be remotely controlled simply by illuminating it with an appropriate wavelength. We show that interactions of conventional impermeable particles remain practically unaffected by light, but, in contrast, for porous particles, we observe a long-range repulsion, several orders of magnitude longer than any conceivable equilibrium surface force. This repulsion emerges due to the diffusio-osmotic flow generated near the porous particles that in this scenario are playing a role of micropumps. The diffusio-osmotic repulsion of porous particles can be used for a remote control of their two-dimensional assemblies at the solid wall, and in particular, we demonstrate that by simply using two different illumination wavelengths it is possible to reversibly switch the state of porous particle dispersion from densely packed surface aggregates to a periodic lattice of particles separated by distances on the order of tens of micrometers.


Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2.
Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model.


The interplay between vaccination and social distancing strategies affects COVID19 population-level outcomes.
Social distancing is an effective population-level mitigation strategy to prevent COVID19 propagation but it does not reduce the number of susceptible individuals and bears severe social consequences-a dire situation that can be overcome with the recently developed vaccines. Although a combination of these interventions should provide greater benefits than their isolated deployment, a mechanistic understanding of the interplay between them is missing. To tackle this challenge we developed an age-structured deterministic model in which vaccines are deployed during the pandemic to individuals who do not show symptoms. The model allows for flexible and dynamic prioritization strategies with shifts between target groups. We find a strong interaction between social distancing and vaccination in their effect on the proportion of hospitalizations. In particular, prioritizing vaccines to elderly (60+) before adults (20-59) is more effective when social distancing is applied to adults or uniformly. In addition, the temporal reproductive number Rt is only affected by vaccines when deployed at sufficiently high rates and in tandem with social distancing. Finally, the same reduction in hospitalization can be achieved via different combination of strategies, giving decision makers flexibility in choosing public health policies. Our study provides insights into the factors that affect vaccination success and provides methodology to test different intervention strategies in a way that will align with ethical guidelines.


Best practicable means (BPM) and as low as reasonably practicable (ALARP) in action at Sellafield.
Operators within the UK nuclear industry are required to employ 'best practicable means' (BPM) to control and minimise radioactive discharges. The requirement for the use of BPM is in order to ensure that doses from discharges are as low as reasonably practicable (ALARP). This facet is increasingly becoming the principal feature of discharge authorisations issued by the Environment Agencies under the Radioactive Substances Act, 1993 (RSA 93), which are enforceable by law. It is important to state at the outset, however, that scientific assessments to date indicate that there are no expectations of environmental harm from authorised discharges, even where those discharges have historically been up to two orders of magnitude higher than current levels. There is also no evidence to indicate that foreseeable future discharges from the Sellafield site will make any contribution to environmental harm.