Directed evolution of O6-alkylguanine-DNA alkyltransferase for efficient labeling of fusion proteins with small molecules in vivo.
We report here the generation of mutants of the human O(6)-alkylguanine-DNA alkyltransferase (hAGT) for the efficient in vivo labeling of fusion proteins with synthetic reporter molecules. Libraries of hAGT were displayed on phage, and mutants capable of efficiently reacting with the inhibitor O(6)-benzylguanine were selected based on their ability to irreversibly transfer the benzyl group to a reactive cysteine residue. Using synthetic O(6)-benzylguanine derivatives, the selected mutant proteins allow for a highly efficient covalent labeling of hAGT fusion proteins in vivo and in vitro with small molecules and therefore should become important tools for studying protein function in living cells. In addition to various applications in proteomics, the selected mutants also yield insight into the interaction of the DNA repair protein hAGT with its inhibitor O(6)-benzylguanine.


Do constrained immigration rates and high β diversity explain contrasting productivity-diversity patterns measured at different scales?
The relationship between productivity and diversity is controversial because of disparity between unimodal and monotonic patterns, especially when occurring simultaneously at different scales. We used stream-side artificial channels to investigate how the availability of a major resource (leaf litter) affected stream invertebrate abundance and diversity at leaf-pack and whole-channel scales. At the larger scale, invertebrate diversity increased monotonically with increasing litter resource density, whereas at the smaller scale the relationship was hump-shaped, in keeping with reports in the literature. This divergence at higher resource levels suggests that multiple mechanisms may be operating. Our results indicate that consistently high species turnover (β diversity) caused the monotonic pattern because of a species-area or "sampling effect" in which new species accumulate with increasing number of samples. The hump-shaped pattern was due to constrained immigration because of a "dilution effect" in which a limited number of immigrants is spread out among the increasing number of available patches. We propose that the relationship between productivity or resource availability and α diversity is generally hump-shaped and the scale-dependent contrast in the relationship only arises where the species pool is large and β diversity is high. Differences in β diversity may, therefore, explain some of the contrasting patterns in the productivity-diversity relationship previously reported.We suggest that continuing immigration by rare taxa is important in sustaining species diversity when productivity is high. The hump-shaped pattern has implications for the impact of anthropogenic ecosystem enrichment on species diversity.


Reproducibility of dual-energy X-ray absorptiometry in the upper extremities in stroke patients.
Bone mineral loss after stroke in affected extremities can increase the prevalence of fractures that interrupt the rehabilitation programme and lower the level of activity. In stroke rehabilitation, precise bone mineral measurement is needed in order to assess the risk of fractures. We defined five regions-of-interest of the upper extremity (proximal humerus, distal forearm, distal radius, distal ulna and hand) consistently, so that the position was comfortable for hemiplegic patients to maintain during the measurement. The aim of this study was to determine whether our method using dual-energy X-ray absorptiometry is useful in hemiplegic patients. Ten normal subjects and 15 hemiplegic patients were studied. In normal subjects the reproducibility of our method was evaluated by repeated measurements of both the right and left sides. In hemiplegic patients the reproducibility was evaluated by repeated measurements of the affected side. The coefficient of variation (CV) values were obtained for BMD (0.7-1.7%) and BMC (0.7-2.3%) in normal subjects, and BMD (1.3-2.1%) and BMC (1.7-5.4%) in hemiplegic patients. The CV for the side-to-side BMD and BMC ratios in the normal subjects ranged from 1.1% to 2.2% and from 1.8% to 3.1%, respectively. Our study shows that the reproducibility of our method is sufficient for application to hemiplegic patients.


Myeloid-derived suppressor cell development is regulated by a STAT/IRF-8 axis.
Myeloid-derived suppressor cells (MDSCs) comprise immature myeloid populations produced in diverse pathologies, including neoplasia. Because MDSCs can impair antitumor immunity, these cells have emerged as a significant barrier to cancer therapy. Although much research has focused on how MDSCs promote tumor progression, it remains unclear how MDSCs develop and why the MDSC response is heavily granulocytic. Given that MDSCs are a manifestation of aberrant myelopoiesis, we hypothesized that MDSCs arise from perturbations in the regulation of interferon regulatory factor-8 (IRF-8), an integral transcriptional component of myeloid differentiation and lineage commitment. Overall, we demonstrated that (a) Irf8-deficient mice generated myeloid populations highly homologous to tumor-induced MDSCs with respect to phenotype, function, and gene expression profiles; (b) IRF-8 overexpression in mice attenuated MDSC accumulation and enhanced immunotherapeutic efficacy; (c) the MDSC-inducing factors G-CSF and GM-CSF facilitated IRF-8 downregulation via STAT3- and STAT5-dependent pathways; and (d) IRF-8 levels in MDSCs of breast cancer patients declined with increasing MDSC frequency, implicating IRF-8 as a negative regulator in human MDSC biology. Together, our results reveal a previously unrecognized role for IRF-8 expression in MDSC subset development, which may provide new avenues to target MDSCs in neoplasia.


Relative roles of climatic suitability and anthropogenic influence in determining the pattern of spread in a global invader.
Because invasive species threaten the integrity of natural ecosystems, a major goal in ecology is to develop predictive models to determine which species may become widespread and where they may invade. Indeed, considerable progress has been made in understanding the factors that influence the local pattern of spread for specific invaders and the factors that are correlated with the number of introduced species that have become established in a given region. However, few studies have examined the relative importance of multiple drivers of invasion success for widespread species at global scales. Here, we use a dataset of >5,000 presence/absence records to examine the interplay between climatic suitability, biotic resistance by native taxa, human-aided dispersal, and human modification of habitats, in shaping the distribution of one of the world's most notorious invasive species, the Argentine ant (Linepithema humile). Climatic suitability and the extent of human modification of habitats are primarily responsible for the distribution of this global invader. However, we also found some evidence for biotic resistance by native communities. Somewhat surprisingly, and despite the often cited importance of propagule pressure as a crucial driver of invasions, metrics of the magnitude of international traded commodities among countries were not related to global distribution patterns. Together, our analyses on the global-scale distribution of this invasive species provide strong evidence for the interplay of biotic and abiotic determinants of spread and also highlight the challenges of limiting the spread and subsequent impact of highly invasive species.


Complete genome sequence of Bacillus amyloliquefaciens subsp. plantarum S499, a rhizobacterium that triggers plant defences and inhibits fungal phytopathogens.
Bacillus amyloliquefaciens subsp. plantarum S499 is a plant beneficial rhizobacterium with a good antagonistic potential against phytopathogens through the release of active secondary metabolites. Moreover, it can induce systemic resistance in plants by producing considerable amounts of surfactins. The complete genome sequence of B. amyloliquefaciens subsp. plantarum S499 includes a circular chromosome of 3,927,922bp and a plasmid of 8,008bp. A remarkable abundance in genomic regions of putative horizontal origin emerged from the analysis. Furthermore, we highlighted the presence of genes involved in the establishment of interactions with the host plants at the root level and in the competition with other soil-borne microorganisms. More specifically, genes related to the synthesis of amylolysin, amylocyclicin, and butirosin were identified. These antimicrobials were not known before to be part of the antibiotic arsenal of the strain. The information embedded in the genome will support the upcoming studies regarding the application of B. amyloliquefaciens isolates as plant-growth promoters and biocontrol agents.


Post-licensure rapid immunization safety monitoring program (PRISM) data characterization.
The Post-Licensure Rapid Immunization Safety Monitoring (PRISM) program is the immunization safety monitoring component of FDA's Mini-Sentinel project, a program to actively monitor the safety of medical products using electronic health information. FDA sought to assess the surveillance capabilities of this large claims-based distributed database for vaccine safety surveillance by characterizing the underlying data. We characterized data available on vaccine exposures in PRISM, estimated how much additional data was gained by matching with select state and local immunization registries, and compared vaccination coverage estimates based on PRISM data with other available data sources. We generated rates of computerized codes representing potential health outcomes relevant to vaccine safety monitoring. Standardized algorithms including ICD-9 codes, number of codes required, exclusion criteria and location of the encounter were used to obtain the background rates. The majority of the vaccines routinely administered to infants, children, adolescents and adults were well captured by claims data. Immunization registry data in up to seven states comprised between 5% and 9% of data for all vaccine categories with the exception of 10% for hepatitis B and 3% and 4% for rotavirus and zoster respectively. Vaccination coverage estimates based on PRISM's computerized data were similar to but lower than coverage estimates from the National Immunization Survey and Healthcare Effectiveness Data and Information Set. For the 25 health outcomes of interest studied, the rates of potential outcomes based on ICD-9 codes were generally higher than rates described in the literature, which are typically clinically confirmed cases. PRISM program's data on vaccine exposures and health outcomes appear complete enough to support robust safety monitoring.


Determining Preferences Related to HIV Counselling and Testing Services Among High School Learners in KwaZulu-Natal: A Discrete Choice Experiment.
A key strategy of the South African national response to HIV is the scale-up of HIV counselling and testing (HCT) in the 15-49 years age group. The integrated school health policy aims to guide the roll out of youth-friendly health services including the provision of HCT in schools. Using a discrete choice experiment to examine preferences regarding the attributes of HCT service packages, this study identifies barriers to and facilitators of HCT among high school learners. Monetary considerations were found to have the strongest effect of any attribute on choice, whilst confidentiality was found to be a primary concern for learners considering HCT. Policy makers and service providers must ensure that confidentiality is maintained, and could consider using monetary incentives as a way of increasing uptake of HCT. Programmes designed to reduce social stigma and improve education and knowledge dissemination around HCT and HIV, are vital in creating demand for HCT and changing attitudes among young people.


Harmonic cut and regularized centroid transform for localization of subcellular structures.
Two novel computational techniques, harmonic cut and regularized centroid transform, are developed for segmentation of cells and their corresponding substructures observed with an epi-fluorescence microscope. Harmonic cut detects small regions that correspond to small subcellular structures. These regions also affect the accuracy of the overall segmentation. They are detected, removed, and interpolated to ensure continuity within each region. We show that interpolation within each region (subcellular compartment) is equivalent to solving the Laplace equation on a multi-connected domain with irregular boundaries. The second technique, referred to as the regularized centroid transform, aims to separate touching compartments. This is achieved by adopting a quadratic model for the shape of the object and relaxing it for final segmentation.


Restricting environmental stimulation influences levels and variability of plasma cortisol.
Restricting stimulation from the environment has been shown to alter psychological and physiological states. The present study of 27 healthy subjects examines the effects of restricted environmental stimulation technique (REST) on plasma levels of cortisol and variability in plasma cortisol levels across repeated REST sessions. The REST environment consisted of a 1.2 X 1.2 X 2.4-m ovoid chamber containing 25 cm of saturated MgSO4 solution (sp gr 1.28) maintained at 34.5 degrees F. The buoyant supinely floating subject experienced a minimum of light, sound, and temperature awareness and spatial orientation. The non-REST environment was a cushioned reclining chair in a quiet dimly lit room. The 5-wk protocol consisted of four visits for blood sampling during a 2-wk baseline followed by eight REST or non-REST sessions, 40 min each, with blood samples taken on four nonsession days between sessions 5 and 8. Variability in plasma cortisol was expressed in terms of standard deviation. REST was associated with across-session decreases of 21.6% in plasma cortisol and 50.5% in plasma cortisol variability, whereas no changes in these measures occurred in non-REST. It is concluded that REST influences both static and dynamic aspects of adrenocortical function, possibly altering the feedback monitoring of plasma cortisol.


MERS-CoV in sheep, goats, and cattle, United Arab Emirates, 2019: Virological and serological investigations reveal an accidental spillover from dromedaries.
The recent COVID-19 pandemic has demonstrated again the global threat posed by emerging zoonotic coronaviruses. During the past two decades alone, humans have experienced the emergence of several coronaviruses, such as SARS-CoV in 2003, MERS-CoV in 2012, and SARS-CoV-2 in 2019. To date, MERS-CoV has been detected in 27 countries, with a case fatality ratio of approximately 34.5%. Similar to other coronaviruses, MERS-CoV presumably originated from bats; however, the main reservoir and primary source of human infections are dromedary camels. Other species within the Camelidae family, such as Bactrian camels, alpacas, and llamas, seem to be susceptible to the infection as well, although to a lesser extent. In contrast, susceptibility studies on sheep, goats, cattle, pigs, chickens, and horses obtained divergent results. In the present study, we tested nasal swabs and/or sera from 55 sheep, 45 goats, and 52 cattle, collected at the largest livestock market in the United Arab Emirates, where dromedaries are also traded, for the presence of MERS-CoV nucleic acid by RT-qPCR, and for specific antibodies by immunofluorescence assay. All sera were negative for MERS-CoV-reactive antibodies, but the nasal swab of one sheep (1.8%) repeatedly tested positive for MERS-CoV nucleic acid. Next generation sequencing (NGS) of the complete N gene of the sheep-derived MERS-CoV revealed >99% nucleotide identity to MERS-CoV sequences of five dromedaries in nearby pens and to three reference sequences. The NGS sequence of the sheep-derived MERS-CoV was confirmed by conventional RT-PCR of a part of the N gene and subsequent Sanger sequencing. All MERS-CoV sequences clustered within clade B, lineage 5. In conclusion, our study shows that noncamelid livestock, such as sheep, goats, and cattle do not play a major role in MERS-CoV epidemiology. The one sheep that tested positive most likely reflects an accidental viral spillover event from infected dromedaries in nearby pens.


Inducible hydrogen sulfide synthesis in chondrocytes and mesenchymal progenitor cells: is H2S a novel cytoprotective mediator in the inflamed joint?
Hydrogen sulfide (H(2)S) has recently been proposed as an endogenous mediator of inflammation and is present in human synovial fluid. This study determined whether primary human articular chondrocytes (HACs) and mesenchymal progenitor cells (MPCs) could synthesize H(2)S in response to pro-inflammatory cytokines relevant to human arthropathies, and to determine the cellular responses to endogenous and pharmacological H(2)S. HACs and MPCs were exposed to IL-1β, IL-6, TNF-α and lipopolysaccharide (LPS). The expression and enzymatic activity of the H(2)S synthesizing enzymes cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) were determined by Western blot and zinc-trap spectrophotometry, respectively. Cellular oxidative stress was induced by H(2)O(2), the peroxynitrite donor SIN-1 and 4-hydroxynonenal (4-HNE). Cell death was assessed by 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Mitochondrial membrane potential (DCm) was determined in situ by flow cytometry. Endogenous H(2) S synthesis was inhibited by siRNA-mediated knockdown of CSE and CBS and pharmacological inhibitors D,L-propargylglycine and aminoxyacetate, respectively. Exogenous H(2)S was generated using GYY4137. Under basal conditions HACs and MPCs expressed CBS and CSE and synthesized H(2)S in a CBS-dependent manner, whereas CSE expression and activity was induced by treatment of cells with IL-1β, TNF-α, IL-6 or LPS. Oxidative stress-induced cell death was significantly inhibited by GYY4137 treatment but increased by pharmacological inhibition of H(2)S synthesis or by CBS/CSE-siRNA treatment. These data suggest CSE is an inducible source of H(2)S in cultured HACs and MPCs. H(2)S may represent a novel endogenous mechanism of cytoprotection in the inflamed joint, suggesting a potential opportunity for therapeutic intervention.


Functional dissociation between medial and lateral prefrontal cortical spatiotemporal activation in negative and positive emotions: a combined fMRI/MEG study.
The orbitofrontal cortex has been cytoarchitectonically and connectionally subdivided into a medial and a lateral part which are assumed to subserve distinct functions in emotional processing. However the exact spatiotemporal mechanisms of negative and positive emotional processing in medial and lateral orbitofrontal cortex remain unclear. We therefore investigated spatiotemporal orbitofrontal and prefrontal cortical activation patterns during emotional stimulation in a combined fMRI/MEG study. We investigated 10 healthy subjects, 5 women and 5 men. Positive and negative pictures from the International Affective Picture system (IAPS) were used for emotional stimulation, whereas neutral and gray pictures were taken as control conditions. fMRI/MEG measurements covered the whole frontal lobe and a time window between -2000 and +200 ms around motor responses (right index finger extension) associated with each picture. Positively and negatively correlated activities were determined in various prefrontal/frontal cortical regions in fMRI. Isocontour maps and single dipoles in MEG were analyzed in 50 ms time windows ranging from -2000 to +200 ms. Dipoles and fMR images were mapped on three-dimensional anatomical MRI so that anatomical localization of single dipoles and regional fMRI activity could be compared. Both negative and positive emotional conditions differed from non-emotional control conditions by strong orbitofrontal and lateral prefrontal activation as well as by the presence of early magnetic fields (-1700 to +1100 ms). Negative emotional processing was characterized by strong medial orbitofrontal activation and earlier (-1700 ms), stronger and more medially oriented orbitofrontal dipoles. In contrast positive emotional processing showed a rather strong activation in lateral prefrontal cortex with later (-1500 ms), weaker and more laterally oriented orbito and prefrontal dipoles. Negative emotional processing can be characterized by strong and early medial orbitofrontal cortical activation, whereas positive emotional processing showed rather later and weaker activation in lateral orbitofrontal/prefrontal cortex. Such a functional dissociation between medial and lateral orbito-frontal/prefrontal cortex during negative and positive emotional processing lends further support to the assumption of a functional subdivision in the orbitofrontal cortex.


Automated segmentation and tracking of non-rigid objects in time-lapse microscopy videos of polymorphonuclear neutrophils.
Time-lapse microscopy is an important technique to study the dynamics of various biological processes. The labor-intensive manual analysis of microscopy videos is increasingly replaced by automated segmentation and tracking methods. These methods are often limited to certain cell morphologies and/or cell stainings. In this paper, we present an automated segmentation and tracking framework that does not have these restrictions. In particular, our framework handles highly variable cell shapes and does not rely on any cell stainings. Our segmentation approach is based on a combination of spatial and temporal image variations to detect moving cells in microscopy videos. This method yields a sensitivity of 99% and a precision of 95% in object detection. The tracking of cells consists of different steps, starting from single-cell tracking based on a nearest-neighbor-approach, detection of cell-cell interactions and splitting of cell clusters, and finally combining tracklets using methods from graph theory. The segmentation and tracking framework was applied to synthetic as well as experimental datasets with varying cell densities implying different numbers of cell-cell interactions. We established a validation framework to measure the performance of our tracking technique. The cell tracking accuracy was found to be >99% for all datasets indicating a high accuracy for connecting the detected cells between different time points.


Exploring fine-scale human and livestock movement in western Kenya.
Human and livestock mobility are key factors in the transmission of several high-burden zoonoses such as rift valley fever and trypanosomiasis, yet our knowledge of this mobility is relatively poor due to difficulty in quantifying population-level movement patterns. Significant variation in the movement patterns of individual hosts means it is necessary to capture their fine-scale mobility in order to gain useful knowledge that can be extrapolated to a population level. Here we explore how the movements of people and their ruminants, and their exposure to various types of land cover, correlate with ruminant ownership and other demographic factors which could affect individual exposure to zoonoses. The study was conducted in Busia County, western Kenya, where the population are mostly subsistence farmers operating a mixed crop/livestock farming system. We used GPS trackers to collect movement data from 26 people and their ruminants for 1 week per individual in July/August 2016, and the study was repeated at the end of the same year to compare movement patterns between the short rainy and dry seasons respectively. We found that during the dry season, people and their ruminants travelled further on trips outside of the household, and that people spent less time on swampland compared to the short rainy season. Our findings also showed that ruminant owners spent longer and travelled further on trips outside the household than non-ruminant owners, and that people and ruminants from poorer households travelled further than people from relatively wealthier households. These results indicate that some individual-level mobility may be predicted by season and by household characteristics such as ruminant ownership and household wealth, which could have practical uses for assessing individual risk of exposure to some zoonoses and for future modelling studies of zoonosis transmission in similar rural areas.


Initiation of regular marijuana use among a cohort of women infected with or at risk for HIV in the Women's Interagency HIV Study (WIHS).
Our study sought to determine the incidence of weekly marijuana use among HIV-infected and uninfected women, to identify correlates of weekly marijuana use, and to test its association with stage of HIV disease and type of HIV treatment received. A total of 2059 HIV-positive and 569 HIV-negative women from 6 sites were recruited between 1994 and 1995 and followed through 2000. After excluding women who reported weekly marijuana use at baseline, 2050 women were included in the analysis. The incidence rate for initiating marijuana was calculated and survival analysis was performed to determine the correlates of initiating weekly marijuana use. Three hundred and three women initiated weekly marijuana use within 5.5 years of the baseline visit, yielding a cumulative incidence (CI) of 14.8%. There was no significant difference in weekly marijuana use initiation between HIV-infected (CI = 14.5%) and HIV-uninfected women (CI = 16.0%). Younger age and having more sex partners was associated with incident weekly marijuana use among both infected and uninfected women. While undetectable viral load was associated with lower incidence rate (p < 0.001, RH = 0.44) and wasting syndrome with higher incidence (p < 0.01, relative hazard [RH] = 3.1), CD4 count was not. Compared to receiving no AIDS treatment at all, women who received basic combination antiretroviral therapy had significantly higher incidence of weekly marijuana use (p < 0.001, RH = 1.93), while highly active antiretroviral therapy (HAART) receivers had significantly lower incidence (p < 0.001, RH = 0.24). In summary, among HIV-infected women, the incidence of weekly marijuana use was associated with only one marker of HIV disease stage and HAART was associated with lower initiation rate of weekly marijuana use.


[Tobacco smoking and mental health].
Smoking continues to be one of the most important health burdens worldwide. To describe smoking habits and associated risk factors in the population of Santiago, Chile. A cross sectional study of a representative sample of the population, from 16 to 64 years old, residents of Santiago, Chile (total population: 3,237,286). A structured interview that included questions about use of tobacco, the CIS-R interviews, used for common mental disorders, were applied. From the sample of 4,693 households, 3,870 people were interviewed (52.2% women, 47.8% men) and 10% refused. Forty percent of the population currently smoked (52.5% men, 47.8% women). "Being a current smoker" was associated with being younger than 55, male sex, and having a common mental disorder. Smoking is highly prevalent in Chile, as compared with developed countries and with some developing countries. Gender differences in use of tobacco have decreased. A higher risk of smoking for people with mental disorders is confirmed.


Mental Health Interventions in the Workplace and Work Outcomes: A Best-Evidence Synthesis of Systematic Reviews.
Mental health issues in the workplace are a growing concern among organizations and policymakers, but it remains unclear what interventions are effective in preventing mental health problems and their associated organizational consequences. This synthesis reports on workplace mental health interventions that impact absenteeism, productivity and financial outcomes. To determine the level of evidence supporting mental health interventions as valuable to work outcomes. Databases were searched for systematic reviews between 2000 and 2012: Medline, EMBASE, the Cochrane Database of Systematic Reviews, DARE, CINAHL, PsycINFO and TRIP. Grey literature searches included health-evidence.ca, Rehab+, National Rehabilitation Information Center (NARIC), and Institute for Work and Health. The assessment of articles for inclusion criteria and methodological quality was conducted independently by two or more researchers, with differences resolved through consensus. The search resulted in 3363 titles, of which 3248 were excluded following title/abstract review, with 115 articles retrieved for full-text review. 14 articles finally met the inclusion criteria and are summarized in this synthesis. There is moderate evidence for the effectiveness of workplace mental health interventions on improved workplace outcomes. Certain types of programs, such as those incorporating both mental and physical health interventions, multicomponent mental health and/or psychosocial interventions, and exposure in vivo containing interventions for particular anxiety disorders had a greater level of research evidence to support their effectiveness.


Ultrashort pulsed laser treatment inactivates viruses by inhibiting viral replication and transcription in the host nucleus.
Ultrashort pulsed laser irradiation is a new method for virus reduction in pharmaceuticals and blood products. Current evidence suggests that ultrashort pulsed laser irradiation inactivates viruses through an impulsive stimulated Raman scattering process, resulting in aggregation of viral capsid proteins. However, the specific functional defect(s) in viruses inactivated in this manner have not been demonstrated. This information is critical for the optimization and the extension of this treatment platform to other applications. Toward this goal, we investigated whether viral internalization, replication, or gene expression in cells were altered by ultrashort pulsed laser irradiation. Murine Cytomegalovirus (MCMV), an enveloped DNA virus, was used as a model virus. Using electron and fluorescence microscopy, we found that laser-treated MCMV virions successfully internalized in cells, as evidenced by the detection of intracellular virions, which was confirmed by the detection of intracellular viral DNA via PCR. Although the viral DNA itself remained polymerase-amplifiable after laser treatment, no viral replication or gene expression was observed in cells infected with laser-treated virus. These results, along with evidence from previous studies, support a model whereby the laser treatment stabilizes the capsid, which inhibits capsid uncoating within cells. By targeting the mechanical properties of viral capsids, ultrashort pulsed laser treatment represents a unique potential strategy to overcome viral mutational escape, with implications for combatting emerging or drug-resistant pathogens.


Two-year results of paclitaxel-eluting stents in patients with medically treated diabetes mellitus from the TAXUS ARRIVE program.
Drug-eluting stents decrease revascularization compared with bare metal stents in diabetic patients, but few studies have compared drug-eluting stent use in diabetic versus nondiabetic patients. The objective of this study was to assess whether paclitaxel provides equivalent revascularization decrease in diabetic and nondiabetic patients. The ARRIVE registries enrolled 7,492 patients receiving TAXUS Express stents, including 2,112 with medically treated diabetes; results were compared with those in the remaining 5,380 nondiabetic patients. Two-year target lesion revascularization (TLR) was comparable in diabetic and nondiabetic patients (8.2% vs 7.7%, p = 0.59) and remained similar after multivariate adjustment for baseline differences (7.1% vs 6.8%, p = 0.41). There were no significant TLR differences between diabetic and nondiabetic patients with small vessels (9.7% vs 9.5%, p = 0.96) or left main coronary artery, 3-vessel, or bifurcation stenting (10.7% vs 13.1%, p = 0.41). Diabetes was not a significant TLR predictor (hazard ratio 0.92, 95% confidence interval 0.77 to 1.12, p = 0.41). Stent thrombosis (2.6% vs 2.4%, p = 0.55) and myocardial infarction (3.8% vs 3.0%, p = 0.09) rates were also similar for diabetic and nondiabetic patients. However, 2-year mortality was significantly increased in diabetic compared with nondiabetic patients (9.7% vs 5.3%, p <0.001). Increased mortality drove significantly increased major cardiac events in diabetics; however, there was no difference in stent-related major cardiac events (8.9% vs 10.1%, p = 0.13). In conclusion, these results suggest that TAXUS paclitaxel-eluting stents abrogate the increased diabetic risk of clinical restenosis previously reported with bare metal stents, with similar low risk of myocardial infarction or stent thrombosis for diabetic and nondiabetic patients. However, diabetic patients still have increased risk of 2-year mortality.


Attitudes towards euthanasia.
There are an infinite variety of attitudes to euthanasia, each individual response to the concept being influenced by many factors. Consequently there is a literature on the subject ranging from the popular article to papers in specialized journals. This study, however, has taken a well defined sample of people, inviting them to answer a questionnaire which was designed to elicit their attitudes to euthanasia in a way which could be analysed statistically. Nor surprisingly attitudes appeared to 'harden' as those answering the questionnaire grew more experienced in dealing with patients and also more professionally established. Thus it was found that of the seven groups questioned practising physicians showed more positive attitudes to euthanasia and their responses did not differ significantly from those of senior medical students. It is these groups which actually or potentially have to resolve the clinical dilemma posed by the dying patient.


Prediction of altered 3'- UTR miRNA-binding sites from RNA-Seq data: the swine leukocyte antigen complex (SLA) as a model region.
THE SLA (swine leukocyte antigen, MHC: SLA) genes are the most important determinants of immune, infectious disease and vaccine response in pigs; several genetic associations with immunity and swine production traits have been reported. However, most of the current knowledge on SLA is limited to gene coding regions. MicroRNAs (miRNAs) are small molecules that post-transcriptionally regulate the expression of a large number of protein-coding genes in metazoans, and are suggested to play important roles in fine-tuning immune mechanisms and disease responses. Polymorphisms in either miRNAs or their gene targets may have a significant impact on gene expression by abolishing, weakening or creating miRNA target sites, possibly leading to phenotypic variation. We explored the impact of variants in the 3'-UTR miRNA target sites of genes within the whole SLA region. The combined predictions by TargetScan, PACMIT and TargetSpy, based on different biological parameters, empowered the identification of miRNA target sites and the discovery of polymorphic miRNA target sites (poly-miRTSs). Predictions for three SLA genes characterized by a different range of sequence variation provided proof of principle for the analysis of poly-miRTSs from a total of 144 M RNA-Seq reads collected from different porcine tissues. Twenty-four novel SNPs were predicted to affect miRNA-binding sites in 19 genes of the SLA region. Seven of these genes (SLA-1, SLA-6, SLA-DQA, SLA-DQB1, SLA-DOA, SLA-DOB and TAP1) are linked to antigen processing and presentation functions, which is reminiscent of associations with disease traits reported for altered miRNA binding to MHC genes in humans. An inverse correlation in expression levels was demonstrated between miRNAs and co-expressed SLA targets by exploiting a published dataset (RNA-Seq and small RNA-Seq) of three porcine tissues. Our results support the resource value of RNA-Seq collections to identify SNPs that may lead to altered miRNA regulation patterns.


A large decline of tropospheric NO2 in China observed from space by SNPP OMPS.
Nitrogen oxides (NOx) are important constituents of air pollution. Here we use retrievals of tropospheric NO2 column from the Ozone Mapping Profiler Suite (OMPS) Nadir Mapper (NM) onboard the Suomi National Polar Partnership (SNPP) spacecraft to analyze global changes of NO2 from 2012 to 2017. The largest decline of NO2 is detected in eastern China, at a rate of -7.3 ± 1.5% per year, and almost entirely driven by wintertime decreases, indicative of decreasing anthropogenic NOx emissions. During the same period, NO2 over other regions around the globe was either stabilizing or changing at a rate only a fraction of that in China. In the case of the US, OMPS reports a statistically insignificant decreasing trend in NO2, consistent with recent work based on the Ozone Monitoring Instrument (OMI), which can be almost entirely attributed to meteorology. OMPS thus offers a continuation of satellite NO2 products from OMI to future JPSS satellites.


Randomized Phase II Study of Azacitidine Alone or in Combination With Lenalidomide or With Vorinostat in Higher-Risk Myelodysplastic Syndromes and Chronic Myelomonocytic Leukemia: North American Intergroup Study SWOG S1117.
Purpose Azacitidine is standard, first-line therapy in higher-risk myelodysplastic syndromes (MDS). Whether azacitidine-based combinations with lenalidomide or vorinostat produce superior overall response rates (ORRs) to azacitidine is not known. Patients and Methods North American Intergroup Study S1117 is a phase II/III trial that randomly assigned patients with higher-risk MDS and chronic myelomonocytic leukemia (CMML) 1:1:1 to azacitidine (75 mg/m2/day on days 1 to 7 of a 28-day cycle); azacitidine plus lenalidomide (10 mg/day on days 1 to 21); or azacitidine plus vorinostat (300 mg twice daily on days 3 to 9). The primary phase II end point was improved ORR. Results Of 277 patients from 90 centers, 92 received azacitidine, 93 received azacitidine plus lenalidomide, and 92 received azacitidine plus vorinostat. Median age was 70 years (range, 28 to 93 years), 85 patients (31%) were female, and 53 patients (19%) had CMML. Serious adverse events were similar across arms, although combination-arm patients were more likely to undergo nonprotocol-defined dose modifications ( P < .001).With a median follow-up of 23 months (range, 1 to 43 months), the ORR was 38% for patients receiving azacitidine, 49% for azacitidine plus lenalidomide ( P = .14 v azacitidine), and 27% for azacitidine plus vorinostat ( P = .16 v azacitidine). For patients with CMML, ORR was higher for azacitidine plus lenalidomide versus azacitidine (68% v 28%, P = .02) but similar for all arms across cytogenetic subgroups, as was remission duration and overall survival. ORR was higher with mutations in DNMT3A and lower for SRSF2, whereas ORR duration improved with fewer mutations. Lenalidomide dose reduction was associated with worse overall survival (hazard ratio, 1.30; P = .05). Conclusion Patients with higher-risk MDS treated with azacitidine-based combinations had similar ORR to azacitidine monotherapy, although patients with CMML benefitted from azacitidine plus lenalidomide. The efficacy of combination regimens may have been affected by dose modifications.


Microstructural analysis and transport properties of MoO and MoC nanostructures prepared by focused electron beam-induced deposition.
By electron-beam-induced deposition, we have succeeded in the direct fabrication of nanowires of molybdenum oxide (MoOx) and molybdenum carbide (MoC) on a SiO2 substrate set in a scanning electron microscope. In order to prepare MoOx specimens of high purity, a precursor gas of molybdenum hexacarbonyl [Mo(CO)6] is used, mixed with oxygen gas. On the other hand, MoC is grown by mixing H2O gas with the precursor gas. The electrical transport properties of the nanowires are investigated by the DC four-terminal method. A highly resistive MoOx nanowire prepared from an as-deposited specimen by annealing in air shows nonlinear current-voltage characteristics and a high photoconductivity. The resistivity ρ of an as-deposited amorphous MoC (a-MoC) nanowire takes its maximum at a temperature T ≈ 10 K and decreases to ≈ 0 with decreasing temperature. This behavior of ρ(T) indicates the possible occurrence of superconductivity in a-MoC nanowires. The characteristic of ρ(T) below the superconducting transition temperature Tc ≈ 4 K can be well explained by the quantum phase-slip model with a coherence length ξ(0) ≈ 8 nm at T = 0.


Minimal genome-wide human CRISPR-Cas9 library.
CRISPR guide RNA libraries have been iteratively improved to provide increasingly efficient reagents, although their large size is a barrier for many applications. We design an optimised minimal genome-wide human CRISPR-Cas9 library (MinLibCas9) by mining existing large-scale gene loss-of-function datasets, resulting in a greater than 42% reduction in size compared to other CRISPR-Cas9 libraries while preserving assay sensitivity and specificity. MinLibCas9 provides backward compatibility with existing datasets, increases the dynamic range of CRISPR-Cas9 screens and extends their application to complex models and assays.


The use of sediment toxicity identification evaluation methods to evaluate clean up targets in an urban estuary.
The Ballona Creek Estuary (BCE) in Los Angeles, California, is in a highly urbanized watershed, is contaminated by a variety of chemicals, and has prevalent sediment toxicity. Sediment cleanup targets for BCE have been established for Cu, Cd, Pb, Zn, chlordane, DDTs, PCBs, and PAHs, based on sediment quality guidelines. A sediment toxicity identification evaluation (TIE) was conducted to examine how these targets corresponded to toxicity observed with the estuarine amphipod Eohaustorius estuarius. Whole sediment and porewater TIEs were used to identify the cause of toxicity. Passive samplers were deployed to determine the bioavailable fraction of contaminants. Spiked sediment tests were conducted to determine the thresholds of toxicity for selected constituents. Toxicity was found to be widespread but temporally and spatially variable. Whole sediment and porewater TIEs both indicated pyrethroid pesticides were the most likely contaminant group contributing to the toxicity. Concentrations of the chemicals listed for cleanup were found to often exceed target values but were not observed at concentrations likely to cause toxicity. Bioavailable fractions of the target chemicals quantified using passive samplers did not exceed toxicity thresholds. Spiked sediment tests established 10 day LC50s for 4,4' DDE, 4, 4' DDT, α-chlordane, and cyfluthrin at >3050 μg/g, 266 μg/g, >2120 μg/g, and 0.33 μg/g organic carbon (OC), respectively. The cyfluthrin LC50 was within the range of concentrations observed in the estuary sediments, but LC50s for the other 3 chemicals were orders of magnitude greater than observed levels. The combination of TIE, sediment chemistry and the results from spiked sediment exposures indicate pyrethroid pesticides are more likely the cause of the observed toxicity than any of the contaminants targeted for cleanup. The results of this study indicate the importance of using a TIE approach to determine chemicals of concern and dose-response information to set cleanup targets, rather than using sediment quality guidelines. Integr Environ Assess Manag 2014;10:260-268. © 2013 SETAC.


The natural history of multiple sclerosis: a geographically based study: 8: familial multiple sclerosis.
We have examined the demographics and long-term outcome of 1044 patients with sporadic and familial multiple sclerosis in a population-based cohort from London, Ontario. The mean follow-up was 25 years in duration, and by this time most patients had reached the unambiguous endpoint scores of the Kurtzke disability status scale (DSS), DSS 6, 8 or 10. An affected family member was identified in 19.8% of the total population, and this subgroup was further divided arbitrarily into the following three groups by the type and number of relatives affected: (i) first degree only; (ii) first degree plus others; (iii) second or third degree. The outcome in these groups was compared with that for those patients who, at a mean 25 year follow-up, had no relatives known to be affected. Familial cases closely resembled those remaining sporadic in both demographics and outcome, although onset in the most heavily loaded families was earlier and male/female ratio was greater. The times to DSS 6, 8 and 10 did not differ significantly when sporadic, familial and familial subgroups were compared. These results provide no clinical support for viewing familial multiple sclerosis as distinct from the sporadic form. The observed recurrence rate for siblings in a strictly defined epidemiological sample was 3.5%, much as projected. These results validate the recurrence risks which have previously been derived from age-corrected data for these first-degree relatives.


A synthetic peptide inhibitor of human immunodeficiency virus replication: correlation between solution structure and viral inhibition.
A peptide designated DP-107 was synthesized containing amino acid residues 558-595 of the envelope glycoprotein gp160 of human immunodeficiency virus type 1 strain LAI (HIV-1LAI). Algorithms for secondary structure have predicted that this region of the envelope transmembrane protein should form an extended alpha-helix. Consistent with this prediction, analysis by circular dichroism (CD) indicated that, under physiological conditions, DP-107 is approximately 85% helical. The high degree of stable secondary structure in a synthetic peptide of this size suggests self-association typical of a coiled coil or leucine zipper. In biological assays, the peptide efficiently blocked virus-mediated cell-cell fusion processes as well as infection of peripheral blood mononuclear cells by both prototypic and primary isolates of HIV-1. A single amino acid substitution in the peptide greatly destabilized its solution structure as measured by CD and abrogated its antiviral activity. An analogue containing a terminal cysteine was oxidized to form a dimer, and this modification lowered the dose required for antiviral effect from 5 to about 1 microgram/ml. These results suggest that both oligomerization and ordered structure are necessary for biological activity. They provide insights also into the role of this region in HIV infection and the potential for development of a new class of antiviral agents.


Design, construction and validation of a portable care system for the daily telerehabiliatation of gait.
When designing a complete system of daily-telerehabilitation it should be borne in mind that properly designed methodologies should be furnished for patients to execute specific motion tasks and for care givers to assess the relevant parameters. Whether in hospital or at home, the system should feature two basic elements: (a) instrumented and walking aids or supports, (b) equipment for the assessment of parameters. Being gait the focus, the idea was to design, construct and validate - as an alternative to the complex and expensive instruments currently used - a simple, portable kit that may be easily interfaced/integrated with the most common mechanical tools used in motion rehabilitation (instrumented walkways, aids, supports), with feedback to both patient for self-monitoring and trainer/therapist (present or remote) for clinical reporting. The proposed system consists of: one step-counter, three couples of photo-emitter detectors, one central unit for collecting and processing the telemetrically transmitted data; a software interface on a dedicated PC and a network adapter. The system has been successfully validated in a clinical application on two groups of 16 subjects at the 1st and 2nd level of the Tinetti test. The degree of acceptance by subjects and care-givers was high. The system was also successfully compared with an Inertial Measurement Unit, a de facto standard. The portable kit can be used with different rehabilitation tools and different ground rugosity. The advantages are: (a) very low costs when compared with optoelectronic solutions and other portable solutions; (b) very high accuracy, also for subjects with imbalance problems; (c) good compatibility with any rehabilitative tool.


Optimal management of Ménière's disease.
Confusion in the nomenclature of Ménière's disease and lack of a standard definition of the disorder until 1995 has hampered accurate assessment of treatment efficacy since the presently defined disorder was first described in 1938. The lack of a widely accepted mechanism of the disease has also delayed the development of rational treatments. Past treatments have focused on relieving elevated pressures in the hydropic ear and more recently on treatment of underlying migraine. Present dietary methods of control include sodium restriction and migraine trigger elimination. Pharmacologic treatments include diuretics, migraine prophylactic medications, histamine analogs, and oral steroids. Surgical procedures include intratympanic steroid perfusion, shunts, and ablative procedures when conservative treatments fail. External pressure devices are also used. Evidence of efficacy is lacking for most interventions other than ablation. At our institution, Ménière's disease is treated as a cerebrovascular disorder. Control of risk factors for cerebrovascular ischemia is combined with treatment of pressure dysfunction in the hydropic ear. Screening for risk factors is performed at presentation. Migraine, dyslipidemia, obesity, diabetes, sleep apnea, hypertension, and atherosclerosis are among the major factors that often require medical management. Migraine prophylactic medications, magnesium supplementation, sodium restriction, migraine trigger elimination, diuretics, anticoagulants, and antihypertensives are among the treatments used initially. Steroids administered orally or intratympanically are used if control is not achieved medically, and ablation remains the definitive treatment in unilateral cases experiencing treatment failure.


Skirt clouds associated with the soufriere eruption of 17 april 1979.
A fortuitous and dramatic photograph of the Soufriere eruption column of 17 April 1979 displays a series of highly structured skirt clouds. The gentle distortion of thin, quasi-horizontal layers of moist air has been documented in meteorological situations. It is proposed that at St. Vincent subhorizontal layers of moist air were intensely deformed by the rapidly rising eruption column and were carried to higher altitudes, where they condensed to form the skirt clouds.


High-risk human papilloma virus infection decreases the frequency of dendritic Langerhans' cells in the human female genital tract.
Dendritic cells (DC) are often arranged in planar layers in tissues with high antigenic exposure, such as skin and mucosae. Providing an en face view, this arrangement optimizes in situ analysis regarding morphology (even of individual dendrites), topographic distribution (regular/clustered) and quantification. The few reports on human genital DC usually utilize single markers and conventional sections, restricting immunolabelling only to cell parts sectioned by the cut. To better assess DC in situ, we labelled epithelial sheets, prepared from fresh cervix biopsies, with antibodies to major histocompatibility complex (MHC)-CII, CD1a and Langerin, revealing (with each of these markers) a dense DC network in a planar-like, regular distribution. Using the hybrid capture system to detect the high-risk mucotropic human papilloma virus (HPV) group, 16 positive and five negative women were studied and the results were compared between these groups. DC frequency per area was substantially reduced (to approximately 50% for the three markers) in samples from all HPV-infected patients compared with samples from controls. Unlike HPV(-) samples, Langerin(+) DC in HPV(+) cervix exhibited a highly accentuated dendritic appearance. We believe this to be the first study using these three DC-restricted markers (Langerin, CD1a and MHC-CII) in cervical epithelial sheets from high-risk HPV(+) donors and also the first study to demonstrate the morphological and quantitative changes triggered by high-risk HPV infection. Cervical DC reduction in early, premalignant high-risk HPV infection might represent viral subversion strategies interfering with efficient antigen handling by the immune system's peripheral sentinels, the DC, perhaps hampering appropriate recruitment and subsequent development of effector (cytotoxic) T cells.


The versatility of median sternotomy in general paediatric surgery.
While common in cardiac surgery, median sternotomy (MS) is rarely required in general paediatric surgery. In the era of advancing endoscopic techniques, sternotomy is perceived as an extremely invasive incision, associated with prolonged postoperative recovery and significant morbidity. We conducted a retrospective chart review of all children undergoing MS for non-cardiac indications between January 2007 and September 2012 and describe the pathology, number of ventilated days, duration of intensive care unit stay and analgesic requirements. Our experience over the past 6 years includes 14 children, aged between 8 months and 13 years. Indications for surgery included penetrating mediastinal trauma (1), anterior and posterior mediastinal masses (4), acquired tracheo-oesophageal fistulas secondary to button battery impaction (2), bronchial foreign bodies (2) and bilateral pulmonary metastases secondary to malignancy (5). The range of postoperative ventilation required was 0 - 34 days (median 1 day) and the range of duration of stay in the paediatric intensive care unit 1 - 39 days (median 4 days). Postoperative analgesia was provided with intravenous or oral paracetamol with or without the addition of morphine. All the children tolerated the procedure well with no sternotomy-related complications on follow-up. This review highlights the variety of conditions in which MS provides unrivalled access to the mediastinum and how well the procedure is tolerated by the paediatric patient, and emphasises the importance of sternotomy being within the armamentarium of access techniques of the general paediatric surgeon.


A cruciform-dumbbell model for inverted dimer formation mediated by inverted repeats.
Small inverted repeats (small palindromes) on plasmids have been shown to mediate a recombinational rearrangement event in Escherichia coli leading to the formation of inverted dimers (giant palindromes). This recombinational rearrangement event is efficient and independent of RecA and RecBCD. In this report, we propose a cruciform-dumbbell model to explain the inverted dimer formation mediated by inverted repeats. In this model, the inverted repeats promote the formation of a DNA cruciform which is processed by an endonuclease into a linear DNA with two hairpin loops at its ends. Upon DNA replication, this linear dumbbell-like DNA is then converted to the inverted dimer. In support of this model, linear dumbbell DNA molecules with unidirectional origin of DNA replication (ColE1 ori ) have been constructed and shown to transform E.coli efficiently resulting in the formation of the inverted dimer. The ability of linear dumbbell DNA to transform E.coli suggests that the terminal loops may be important in bypassing the requirement of DNA supercoiling for initiation of replication of the ColE1 ori.


Root cause analysis of non-infectious transfusion complications and the lessons learnt.
Transfusion of blood and blood products can be associated with hazards which may be at times fatal. Timely reporting of transfusion reactions is imperative for root cause analysis and their prevention in future. We retrospectively reviewed the transfusion reactions at our institution during last seven years. The data was retrieved from our computerized blood bank information system and by reviewing the medical charts of patients. The frequency of adverse effects, implicated products, wrong blood transfusion and its outcome were observed. During study period (2006-2012), a total of 393,662 blood or blood products were transfused. There were 458 adverse events with an estimated rate of 1.16 per 1000 blood products administered. During 2011-2012, 121 transfusion reactions were reported of 119,921 transfused units. The most common adverse effects were allergic reactions (70 episodes of 121 or 57.8%) followed by febrile non hemolytic transfusion reactions or FNHTR (43 events of 121 or 35.5%). Transfusion associated dyspnea, circulatory overload and transfusion associated lung injury were less frequent. During the study period, 142,066 red cell units were transfused with nine recognized ABO-mismatch transfusions and two fatalities. The computed incidence of ABO-mismatch transfusion was 1 in 15,785 with a mortality rate of 1 in 71,033 units transfused. Etiology included: errors in final bed side check (n=5), blood bank clerical errors (n=3) and mislabeled tube (n=1). A review of these cases prompted hospital transfusion committee for re-enforcing policies and protocols to minimize accidental ABO incompatible transfusions. We concluded that urticaria and FNHTR are the most frequent transfusion reactions in our setting. ABO mismatched blood transfusions are rare but preventable errors and result mainly from clerical imprecisions.


Vitamin B12 deficiency in relation to functional disabilities.
This study was designed to assess whether symptoms, functional measures, and reported disabilities were associated with vitamin B12 (B12) deficiency when defined in three ways. Participants, aged 60 or more years of age, in 1999-2002 National Health and Nutrition Examination Surveys (NHANES) were categorized in relation to three previously used definitions of B12 deficiency: (1) serum B12 < 148 pmol/L; (2) serum B12 < 200 pmol/L and serum homocysteine > 20 μmol/L; and (3) serum B12 < 258 pmol/L or serum methylmalonic acid > 0.21 μmol/L. Functional measures of peripheral neuropathy, balance, cognitive function, gait speed, along with self-reported disability (including activities of daily living) were examined with standardized instruments by trained NHANES interviewers and technicians. Individuals identified as B12 deficient by definition 2 were more likely to manifest peripheral neuropathy OR (odds) (95% confidence intervals), p value: 9.70 (2.24, 42.07), 0.004 and report greater total disability, 19.61 (6.22, 61.86) 0.0001 after adjustments for age, sex, race, serum creatinine, and ferritin concentrations, smoking, diabetes, and peripheral artery disease. Smaller, but significantly increased, odds of peripheral neuropathy and total disability were also observed when definition 3 was applied. Functional measures and reported disabilities were associated with B12 deficiency definitions that include B12 biomarkers (homocysteine or methylmalonic acid). Further study of these definitions is needed to alert clinicians of possible subclinical B12 deficiency because functional decline amongst older adults may be correctable if the individual is B12 replete.


Quantitative Comparison of the Anterior-Posterior Patterning System in the Embryos of Five Drosophila Species.
Complex spatiotemporal gene expression patterns direct the development of the fertilized egg into an adult animal. Comparisons across species show that, in spite of changes in the underlying regulatory DNA sequence, developmental programs can be maintained across millions of years of evolution. Reciprocally, changes in gene expression can be used to generate morphological novelty. Distinguishing between changes in regulatory DNA that lead to changes in gene expression and those that do not is therefore a central goal of evolutionary developmental biology. Quantitative, spatially-resolved measurements of developmental gene expression patterns play a crucial role in this goal, enabling the detection of subtle phenotypic differences between species and the development of computations models that link the sequence of regulatory DNA to expression patterns. Here we report the generation of two atlases of cellular resolution gene expression measurements for the primary anterior-posterior patterning genes in Drosophila simulans and Drosophila virilis By combining these data sets with existing atlases for three other Drosophila species, we detect subtle differences in the gene expression patterns and dynamics driving the highly conserved axis patterning system and delineate inter-species differences in the embryonic morphology. These data sets will be a resource for future modeling studies of the evolution of developmental gene regulatory networks.


Kleine-Levin syndrome.
Kleine-Levin syndrome, sometimes referred to as Rip van Winkle disease, is a rare sleep disorder mainly affecting teenage boys in which the main features are intermittent hypersomnolence, behavioural and cognitive disturbances, hyperphagia and in some cases hypersexuality. Each episode lasts for one or two weeks, and affected people are entirely asymptomatic between episodes. No definite cause has been identified but hypothalamic dysfunction seems likely. Relapses may occur every few weeks or months, and the condition may last for a decade or more before spontaneous resolution. There is no effective treatment but stimulants such as methylphenidate and modafinil as well as the mood stabiliser lithium carbonate have been tried with varying success.


Pivotal role of the renin/prorenin receptor in angiotensin II production and cellular responses to renin.
Renin is an aspartyl protease essential for the control of blood pressure and was long suspected to have cellular receptors. We report the expression cloning of the human renin receptor complementary DNA encoding a 350-amino acid protein with a single transmembrane domain and no homology with any known membrane protein. Transfected cells stably expressing the receptor showed renin- and prorenin-specific binding. The binding of renin induced a fourfold increase of the catalytic efficiency of angiotensinogen conversion to angiotensin I and induced an intracellular signal with phosphorylation of serine and tyrosine residues associated to an activation of MAP kinases ERK1 and ERK2. High levels of the receptor mRNA are detected in the heart, brain, placenta, and lower levels in the kidney and liver. By confocal microscopy the receptor is localized in the mesangium of glomeruli and in the subendothelium of coronary and kidney artery, associated to smooth muscle cells and colocalized with renin. The renin receptor is the first described for an aspartyl protease. This discovery emphasizes the role of the cell surface in angiotensin II generation and opens new perspectives on the tissue renin-angiotensin system and on renin effects independent of angiotensin II.


Persistence across Pleistocene ice ages in Mediterranean and extra-Mediterranean refugia: phylogeographic insights from the common wall lizard.
Pleistocene climatic oscillations have played a major role in structuring present-day biodiversity. The southern Mediterranean peninsulas have long been recognized as major glacial refugia, from where Northern Europe was post-glacially colonized. However, recent studies have unravelled numerous additional refugia also in northern regions. We investigated the phylogeographic pattern of the widespread Western Palaearctic lizard Podarcis muralis, using a range-wide multilocus approach, to evaluate whether it is concordant with a recent expansion from southern glacial refugia or alternatively from a combination of Mediterranean and northern refugia. We analyzed DNA sequences of two mitochondrial (cytb and nd4) and three nuclear (acm4, mc1r, and pdc) gene fragments in individuals from 52 localities across the species range, using phylogenetic and phylogeographic methods. The complex phylogeographic pattern observed, with 23 reciprocally monophyletic allo- parapatric lineages having a Pleistocene divergence, suggests a scenario of long-term isolation in multiple ice-age refugia across the species distribution range. Multiple lineages were identified within the three Mediterranean peninsulas - Iberia, Italy and the Balkans - where the highest genetic diversity was observed. Such an unprecedented phylogeographic pattern - here called "refugia within all refugia" - compasses the classical scenario of multiple southern refugia. However, unlike the southern refugia model, various distinct lineages were also found in northern regions, suggesting that additional refugia in France, Northern Italy, Eastern Alps and Central Balkans allowed the long-term persistence of this species throughout Pleistocene glaciations. The phylogeography of Podarcis muralis provides a paradigm of temperate species survival in Mediterranean and extra-Mediterranean glacial refugia. Such refugia acted as independent biogeographic compartments for the long-term persistence of this species, for the differentiation of its genetic lineages, and for the short-distance post-glacial re-colonization of neighbouring areas. This finding echoes previous findings from recent phylogeographic studies on species from temperate ecoregions, thus suggesting the need for a reappraisal of the role of northern refugia for glacial persistence and post-glacial assembly of Holarctic biota.


Short-term changes on MRI predict long-term changes on radiography in rheumatoid arthritis: an analysis by an OMERACT Task Force of pooled data from four randomised controlled trials.
In rheumatoid arthritis (RA), MRI provides earlier detection of structural damage than radiography (X-ray) and more sensitive detection of intra-articular inflammation than clinical examination. This analysis was designed to evaluate the ability of early MRI findings to predict subsequent structural damage by X-ray. Pooled data from four randomised controlled trials (RCTs) involving 1022 RA hands and wrists in early and established RA were analysed. X-rays were scored using van der Heijde-modified or Genant-modified Sharp methods. MRIs were scored using Outcome Measures in Rheumatology (OMERACT) RA MRI Score (RAMRIS). Data were analysed at the patient level using multivariable logistic regression and receiver operating characteristic curve analyses. Progression of MRI erosion scores at Weeks 12 and 24 predicted progression of X-ray erosions at Weeks 24 and 52, with areas under the curve (AUCs) of 0.64 and 0.74, respectively. 12-week and 24-week changes in MRI osteitis scores were similarly predictive of 24-week and 52-week X-ray erosion progressions; pooled AUCs were 0.78 and 0.77, respectively. MRI changes in synovitis at Weeks 12 and 24 also predicted progression of X-ray joint damage (erosion and joint-space narrowing) at Weeks 24 and 52 (AUCs=0.72 and 0.65, respectively). Early changes in joint damage and inflammation detected with MRI predict changes in joint damage evident on subsequent X-rays. These findings support the use of MRI as a valid method for monitoring structural damage in short-duration RCTs.


Neutralizing antiviral B cell responses.
Neutralizing antiviral B cell responses differ in various aspects from the many usually measured B cell responses specific for protein in adjuvants. In particular, such neutralizing antiviral B cell responses are more rapidly induced, reach higher titers, are longer lived, and are efficiently generated without adjuvants. Evidence is summarized here that the repetitiveness of many viral antigens is a key factor responsible for the efficiency of these B cell responses, amplifying B cells early and rapidly for potent IgM responses and also for efficient switching to IgG. The data reviewed indicate that B cells discriminate antigen patterns via the degree of surface Ig-cross-linking and use antigen repetitiveness as a self/nonself discriminator.


Analysis of lamprey clustered Fox genes: insight into Fox gene evolution and expression in vertebrates.
In the human genome, members of the FoxC, FoxF, FoxL1, and FoxQ1 gene families are found in two paralagous clusters. One cluster contains the genes FOXQ1, FOXF2, FOXC1 and the second consists of FOXF1, FOXC2, and FOXL1. In jawed vertebrates these genes are known to be expressed in different pharyngeal tissues and all, except FoxQ1, are involved in patterning the early embryonic mesoderm. We have previously traced the evolution of this cluster in the bony vertebrates, and the gene content is identical in the dogfish, a member of the most basally branching lineage of the jawed vertebrates. Here we extend these analyses to jawless vertebrates. Using genomic searches and molecular approaches we have identified homologues of these genes from lampreys. We identify two FoxC genes, two FoxF genes, two FoxQ1 genes and single FoxL1 gene. We examine the embryonic expression of one predominantly mesodermally expressed gene family, FoxC, and the endodermally expressed member of the cluster, FoxQ1. We identified FoxQ1 transcripts in the pharyngeal endoderm, while the two FoxC genes are differentially expressed in the pharyngeal mesenchyme and ectoderm. Furthermore we identify conserved expression of lamprey FoxC genes in the paraxial and intermediate mesoderms. We interpret our results through a chordate-wide comparison of expression patterns and discuss gene content in the context of theories on the evolution of the vertebrate genome.


Long-chain GM1 gangliosides alter transmembrane domain registration through interdigitation.
Extracellular and cytosolic leaflets in cellular membranes are distinctly different in lipid composition, yet they contribute together to signaling across the membranes. Here we consider a mechanism based on long-chain gangliosides for coupling the extracellular and cytosolic membrane leaflets together. Based on atomistic molecular dynamics simulations, we find that long-chain GM1 in the extracellular leaflet exhibits a strong tendency to protrude into the opposing bilayer leaflet. This interdigitation modulates the order in the cytosolic monolayer and thereby strengthens the interaction and coupling across a membrane. Coarse-grained simulations probing longer time scales in large membrane systems indicate that GM1 in the extracellular leaflet modulates the phase behavior in the cytosolic monolayer. While short-chain GM1 maintains phase-symmetric bilayers with a strong membrane registration effect, the situation is altered with long-chain GM1. Here, the significant interdigitation induced by long-chain GM1 modulates the behavior in the cytosolic GM1-free leaflet, weakening and slowing down the membrane registration process. The observed physical interaction mechanism provides a possible means to mediate or foster transmembrane communication associated with signal transduction.


Bench-to-bedside review: Permissive hypercapnia.
Current protective lung ventilation strategies commonly involve hypercapnia. This approach has resulted in an increase in the clinical acceptability of elevated carbon dioxide tension, with hypoventilation and hypercapnia 'permitted' in order to avoid the deleterious effects of high lung stretch. Advances in our understanding of the biology of hypercapnia have prompted consideration of the potential for hypercapnia to play an active role in the pathogenesis of inflammation and tissue injury. In fact, hypercapnia may protect against lung and systemic organ injury independently of ventilator strategy. However, there are no clinical data evaluating the direct effects of hypercapnia per se in acute lung injury. This article reviews the current clinical status of permissive hypercapnia, discusses insights gained to date from basic scientific studies of hypercapnia and acidosis, identifies key unresolved concerns regarding hypercapnia, and considers the potential clinical implications for the management of patients with acute lung injury.


Nonthermal effects of mobile-phone frequency microwaves on uteroplacental functions in pregnant rats.
Exposure to high-density microwaves can cause detrimental effects on the testis, eye, and other tissues, and induce significant biologic changes through thermal actions. To examine nonthermal effect of continuous wave (CW) 915MHz microwaves used in cellular phones, we compared the effects of microwaves with those of heat. Thirty-six pregnant rats were assigned to six groups: rats exposed to microwaves at 0.6 or 3mW/cm(2) incident power density at 915MHz for 90min, rats immersed in water at 38 or 40 degrees C, which induces about the same increase in colonic temperature of 1.0 or 3.5 degrees C as 0.6 or 3mW/cm(2) microwaves, respectively; rats immersed in water at 34 degrees C, which is considered to be thermoneutral; and control rats. We identified significant differences in the uteroplacental circulation, and in placental endocrine and immune functions between pregnant rats immersed in water at 34 and 38 degrees C, but not between rats immersed at 38 degrees C and those exposed to microwaves at 0.6mW/cm(2). By contrast, we observed significant decreases in uteroplacental blood flow and estradiol in rats exposed to microwaves at 3mW/cm(2) as compared with those immersed in water at 40 degrees C. These results suggest microwaves at 0.6mW/cm(2) at 915MHz, equal to a specific absorption rate (SAR) of 0.4W/kg, which is the maximum permissible exposure level recommended by the American National Standards Institute (ANSI), do not exert nonthermal effects on blood estradiol and progesterone, on splenic natural killer cell activity, on the uteroplacental circulation.


Identification and classification of Lactobacillus acidophilus, L. gasseri and L. johnsonii strains by SDS-PAGE and rRNA-targeted oligonucleotide probe hybridization.
Thirty-two strains originally identified as Lactobacillus acidophilus and L. gasseri were screened for their taxonomic homogeneity by SDS-PAGE of whole-cell proteins. After numerical comparison of the resulting protein electrophoretic fingerprints, two well-delineated clusters were detected. The majority of the strains grouped in one electrophoretic cluster, which contained the type strain of L. acidophilus and corresponds to DNA group A1 of Johnson, J. L., Phelps, C. F., Cummins, C. S., London, J. & Gasser, F. (1980; International Journal of Systematic Bacteriology 30, 53-68). Another cluster corresponded to DNA group B. It contained two subclusters, which agreed perfectly with DNA subgroups B1 (L. gasseri) and B2 (L. johnsonii), respectively. The 23S rRNA genes were partially sequenced and 23S-rRNA-targeted oligonucleotide probes were designed for identification of DNA groups A1, B1 and B2. Probe Lbg reacted with all strains of electrophoretic cluster B1 (L. gasseri), probe Lbj hybridized with strains of cluster B2 (L. johnsonii) and probe Lba with strains of cluster A1 (authentic L. acidophilus). The probes were successfully used for the identification of strains belonging to the respective species. The phylogenetic relationship of a representative of L. johnsonii was determined by comparative sequence analysis of the 16S rRNA genes. It is very closely related to L. gasseri.


Evolution of the hominoid vertebral column: The long and the short of it.
The postcranial axial skeleton exhibits considerable morphological and functional diversity among living primates. Particularly striking are the derived features in hominoids that distinguish them from most other primates and mammals. In contrast to the primitive catarrhine morphotype, which presumably possessed an external (protruding) tail and emphasized more pronograde trunk posture, all living hominoids are characterized by the absence of an external tail and adaptations to orthograde trunk posture. Moreover, modern humans evolved unique vertebral features that satisfy the demands of balancing an upright torso over the hind limbs during habitual terrestrial bipedalism. Our ability to identify the evolutionary timing and understand the functional and phylogenetic significance of these fundamental changes in postcranial axial skeletal anatomy in the hominoid fossil record is key to reconstructing ancestral hominoid patterns and retracing the evolutionary pathways that led to living apes and modern humans. Here, we provide an overview of what is known about evolution of the hominoid vertebral column, focusing on the currently available anatomical evidence of three major transitions: tail loss and adaptations to orthograde posture and bipedal locomotion.


Construction of ferrocene modified conducting polymer based amperometric urea biosensor.
Herein, an electrochemical urea sensing bio-electrode is reported that has been constructed by firstly electropolymerizing 4-(2,5-Di(thiophen-2-yl)-1H-pyrrol-1-yl)aniline monomer (SNS-Aniline) on Pencil Graphite Electrode (PGE), then modifying the polymer coated electrode surface with di-amino-Ferrocene (DAFc) as the mediator, and lastly Urease enzyme through glutaraldehyde crosslinking. The effect of pH, temperature, polymer thickness, and applied potential on the electrode current response was investigated besides performing storage and operational stability experiments with the interference studies. The resulting urea biosensor's amperometric response was linear in the range of 0.1-8.5mM with the sensitivity of 0.54μA/mM, detection limit of 12μM, and short response time of 2s. The designed bio-electrode was tested with real human blood and urine samples where it showed excellent analytical performance with insignificant interference.


Plaster Burn: Challenge to Plastic Surgeon.
The thermal burn is accidental and also is the hidden and worst complication of medical plaster application. This study evaluated clinical and aetiological profile and severity of plaster burns. In Department of Burns, Plastic and Reconstructive Surgery from 1st August 2014 to 31st December 2015, six patients with plaster burn were assessed for total body surface area and depth of burn. The wounds were cultured and dressed with moist dressings daily till the surgical procedure and satisfactory healing. The youngest patient was 10 and oldest 65 years (mean age: 40.20±6.67 years, 4 males and two females with ratio of 2:1). Three patients sustained plaster burn injury accidentally at home and 3 developed burn after medical application of plaster. The hands burns were observed commonly in accidental plaster burns, while ankle was often involved in iatrogenic plaster burns. The iatrogenic burns were mostly deep in thickness varying 2nd to 4th degree, while accidental burns were often 2nd degree. Superficial 2nd degree burns were managed conservatively, and deep 2nd degree burns were skin grafted. Fourth degree burn was managed with reverse sural flap alone and another with vacuum-assisted closure followed by reverse sural flap and skin grafting. Plaster burn is still a challenge to plastic surgeon and it is advised for applying casts to utilize all available means to limit the amount of casting material provided. It can be accomplished without compromising the cast strength to minimize the risk of thermal injury when applying plaster or composite casts.


A framework for significance analysis of gene expression data using dimension reduction methods.
The most popular methods for significance analysis on microarray data are well suited to find genes differentially expressed across predefined categories. However, identification of features that correlate with continuous dependent variables is more difficult using these methods, and long lists of significant genes returned are not easily probed for co-regulations and dependencies. Dimension reduction methods are much used in the microarray literature for classification or for obtaining low-dimensional representations of data sets. These methods have an additional interpretation strength that is often not fully exploited when expression data are analysed. In addition, significance analysis may be performed directly on the model parameters to find genes that are important for any number of categorical or continuous responses. We introduce a general scheme for analysis of expression data that combines significance testing with the interpretative advantages of the dimension reduction methods. This approach is applicable both for explorative analysis and for classification and regression problems. Three public data sets are analysed. One is used for classification, one contains spiked-in transcripts of known concentrations, and one represents a regression problem with several measured responses. Model-based significance analysis is performed using a modified version of Hotelling's T2-test, and a false discovery rate significance level is estimated by resampling. Our results show that underlying biological phenomena and unknown relationships in the data can be detected by a simple visual interpretation of the model parameters. It is also found that measured phenotypic responses may model the expression data more accurately than if the design-parameters are used as input. For the classification data, our method finds much the same genes as the standard methods, in addition to some extra which are shown to be biologically relevant. The list of spiked-in genes is also reproduced with high accuracy. The dimension reduction methods are versatile tools that may also be used for significance testing. Visual inspection of model components is useful for interpretation, and the methodology is the same whether the goal is classification, prediction of responses, feature selection or exploration of a data set. The presented framework is conceptually and algorithmically simple, and a Matlab toolbox (Mathworks Inc, USA) is supplemented.


Demonstration of respiratory pigments in clonal insulin-producing beta-cells.
The amounts of cytochromes and flavoproteins in insulin-producing RINm5F-cells were investigated with dual wavelength spectroscopy. The cytochromes a, a3, b and c were present in normal stoichiometric proportions. Whereas each cytochrome occurred in a concentration corresponding to 25-75 mumol/kg protein, the total amount of flavoproteins was equivalent to about 200 mumol/kg protein. In support of the existence of a functional respiratory chain, the cytochromes reacted normally both to additions of oxygen and to the inhibitors antimycin A and amytal.


Fibroblast Growth Factor 23 Trajectories in Chronic Hemodialysis Patients: Lessons from the HEMO Study.
Long-term patterns of fibroblast growth factor 23 (FGF23) are poorly characterized among dialysis patients. To identify different FGF23 trajectories and determine clinical factors that predict distinct FGF23 trajectories and whether FGF23 trajectories differ in regard to their associations with all-cause mortality among prevalent hemodialysis patients. The HEMO study was a randomized multicenter study evaluating the effects of high-dose vs. standard-dose and high-flux vs. low-flux hemodialysis on mortality. We measured intact FGF23 levels in stored serum samples at baseline and annually among 919 HEMO participants and identified FGF23 trajectories using group-based modeling. Logistic regression determined predictors of trajectories. Cox regression models evaluated the association between trajectory and all-cause mortality. We identified 5 distinct FGF23 trajectory groups during the initial 24 months: low stable, low increasing, elevated increasing, elevated decreasing, and elevated stable. In multivariable models, diabetes, high dose dialysis, no venous catheter, low serum calcium, phosphorus, and interleukin-6, no vitamin D analog use, and greater residual kidney function were associated with the low stable trajectory group compared to the elevated stable group. High flux dialysis, no venous catheter, and low serum phosphorus and 25-hydroxyvitamin D were associated with the elevated decreasing trajectory group compared to the elevated stable group. After full adjustment, the low stable trajectory group was associated with reduced mortality (hazard ratio [HR] 0.61; 95% CI -0.41-0.91) compared to the elevated stable trajectory group. We identified 5 distinct FGF23 trajectories over 24 months among HEMO study participants including a decreasing trajectory. The low stable FGF23 trajectory was associated with a reduced HR of all-cause mortality.


Research priorities in the management of severe Plasmodium falciparum malaria in children.
Severe malaria is a common reason for admission to paediatric wards in hospitals across sub-Saharan Africa. Despite over 100 years of research, mortality remains high. Deaths are associated with severe metabolic acidosis, shock, severe anaemia, hypoglycaemia, impaired consciousness, raised intracranial pressure, and status epilepticus. Most inpatient deaths occur within 24 h of admission to hospital, before the beneficial effects of treatment with antimalarial drugs are achieved. This review covers the priority areas for research in the care of children with severe malaria, addressing each of the main risk factors associated with death, in a bid to reduce the inpatient mortality.


Determining best methods to screen for religious/spiritual distress.
This study sought to validate for the first time a brief screening measure for religious/spiritual (R/S) distress given the Commission on Cancer's mandated screening for psychosocial distress including spiritual distress. Data were collected in conjunction with an annual survey of adult hematopoietic cell transplantation (HCT) survivors. Six R/S distress screeners were compared to the Brief RCOPE, Negative Religious Coping subscale as the reference standard. We pre-specified validity as a sensitivity score of at least 85 %. As no individual measure attained this, two post hoc analyses were conducted: analysis of participants within 2 years of transplantation and of a simultaneous pairing of items. Data were analyzed from 1449 respondents whose time since HCT was 6 months to 40 years. For the various single-item screening protocols, sensitivity ranged from 27 (spiritual/religious concerns) to 60 % (meaning/joy) in the full sample and 25 (spiritual/religious concerns) to 65 % (meaning/joy) in a subsample of those within 2 years of HCT. The paired items of low meaning/joy and self-described R/S struggle attained a net sensitivity of 82 % in the full sample and of 87 % in those within 2 years of HCT but with low net specificities. While no single-item screener was acceptable using our pre-specified sensitivity value of 85 %, the simultaneous use of meaning/joy and self-described struggle items among cancer survivors is currently the best choice to briefly screen for R/S distress. Future research should validate this and other approaches in active treatment cancer patients and survivors and determine the best times to screen.


Usefulness of repeated N-terminal pro-B-type natriuretic peptide measurements as incremental predictor for long-term cardiovascular outcome after vascular surgery.
Plasma N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) levels improve preoperative cardiac risk stratification in vascular surgery patients. However, single preoperative measurements of NT-pro-BNP cannot take into account the hemodynamic stress caused by anesthesia and surgery. Therefore, the aim of the present study was to assess the incremental predictive value of changes in NT-pro-BNP during the perioperative period for long-term cardiac mortality. Detailed cardiac histories, rest left ventricular echocardiography, and NT-pro-BNP levels were obtained in 144 patients before vascular surgery and before discharge. The study end point was the occurrence of cardiovascular death during a median follow-up period of 13 months (interquartile range 5 to 20). Preoperatively, the median NT-pro-BNP level in the study population was 314 pg/ml (interquartile range 136 to 1,351), which increased to a median level of 1,505 pg/ml (interquartile range 404 to 6,453) before discharge. During the follow-up period, 29 patients (20%) died, 27 (93%) from cardiovascular causes. The median difference in NT-pro-BNP in the survivors was 665 pg/ml, compared to 5,336 pg/ml in the patients who died (p = 0.01). Multivariate Cox regression analyses, adjusted for cardiac history and cardiovascular risk factors (age, angina pectoris, myocardial infarction, stroke, diabetes mellitus, renal dysfunction, body mass index, type of surgery and the left ventricular ejection fraction), demonstrated that the difference in NT-pro-BNP level between pre- and postoperative measurement was the strongest independent predictor of cardiac outcome (hazard ratio 3.06, 95% confidence interval 1.36 to 6.91). In conclusion, the change in NT-pro-BNP, indicated by repeated measurements before surgery and before discharge is the strongest predictor of cardiac outcomes in patients who undergo vascular surgery.


Molecular Mechanisms in Amyotrophic Lateral Sclerosis: The Role of Angiogenin, a Secreted RNase.
Amyotrophic lateral sclerosis is a fatal neurodegenerative disease caused by the loss of motoneurons. The precise molecular and cellular basis for neuronal death is not yet well established, but the contemporary view is that it is a culmination of multiple aberrant biological processes. Among the proposed mechanisms of motoneuron degeneration, alterations in the homeostasis of RNA binding proteins (RBP) and the consequent changes in RNA metabolism have received attention recently. The ribonuclease, angiogenin was one of the first RBPs associated with familial and sporadic ALS. It is enriched in motoneurons under physiological conditions, and is required for motoneuron survival under stress conditions. Furthermore, delivery of angiogenin protects cultured motoneurons against stress-induced injury, and significantly increases the survival of motoneurons in SOD(G93A) mice. In this overview on the role of angiogenin in RNA metabolism and in the control of motoneuron survival, we discuss potential pathogenic mechanisms of angiogenin dysfunction relevant to ALS and other neurodegenerative disorders. We also discuss recent evidence demonstrating that angiogenin secreted from stressed motoneurons may alter RNA metabolism in astrocytes.


New delivery systems in contraception: vaginal rings.
Vaginal rings, made of soft flexible silicone rubber, for delivery of contraceptive hormones are currently gaining clinical acceptance. This method provides extended release of hormones, which are implanted in the core of the ring and slowly disseminate into vaginal tissue. Although formulations and sizes vary, most rings are approximately 58 mm in diameter with a cross-section of 8.4 mm. Depending on the type of ring used, prolonged hormone release may occur from 3 weeks to 1 year. Advantages of the vaginal ring method are that it is user controlled, does not interfere with intercourse, does not require daily intake of a pill, and allows continuous delivery of a low dose of steroids. The Population Council has developed a progesterone-releasing ring, which is currently on the market in Chile and Peru for contraception in breastfeeding women. This ring may be effective for progesterone therapy during in vitro fertilization as well. A contraceptive ring releasing very low doses of the potent progestin Nestorone (Population Council, New York, NY) for 6 to 12 months is also under investigation. The optimal ring formulations, however, contain hormone combinations that provide excellent contraceptive efficacy with few side effects and good control of menstrual bleeding. The Food and Drug Administration has recently approved Organon's monthly ring releasing etonogestrel and ethinyl estradiol. The Population Council is developing a 1-year contraceptive ring releasing low doses of Nestorone and ethinyl estradiol. Combination rings are associated with very low pregnancy rates and side effects consistent with those of oral contraceptives.


A comparison of the effect of universal use of gloves and gowns with that of glove use alone on acquisition of vancomycin-resistant enterococci in a medical intensive care unit.
To determine the efficacy of the use of gloves and gowns compared with that of the use of gloves alone for the prevention of nosocomial transmission of vancomycin-resistant enterococci. Epidemiologic study and controlled, nonrandomized clinical trial. University-affiliated, 900-bed, urban teaching hospital in which vancomycin-resistant enterococci are endemic. 181 consecutive patients admitted to the medical intensive care unit for 48 hours or more. It was determined that all hospital employees would always use gloves and gowns when attending 8 particular beds in the medical intensive care unit and would always use gloves alone when attending 8 others. Compliance with precautions was monitored weekly. Rectal surveillance cultures were taken from patients daily. Cultures of environmental surfaces, such as those of bed rails, bedside tables, and other frequently touched objects in patient rooms and common areas, were taken monthly. Pulsed-field gel electrophoresis was used for molecular epidemiologic typing of vancomycin-resistant enterococci. The number of patients becoming colonized by vancomycin-resistant enterococci; the number of days to acquisition of vancomycin-resistant enterococci; and other measurements, including nosocomial infections, length of hospital stay, and mortality rates. The 93 patients in glove-and-gown rooms and the 88 patients in glove-only rooms had similar demographic and clinical characteristics. Fifteen (16.1%) patients in the glove-and-gown group and 13 (14.8%) in the glove-only group had vancomycin-resistant enterococci on admission to the medical intensive care unit. Twenty-four (25.8%) patients in the glove-and-gown group and 21 (23.9%) in the glove-only group acquired vancomycin-resistant enterococci in the medical intensive care unit. The mean times to colonization among the patients who became colonized were 8.0 days in the glove-and-gown group and 7.1 days in the glove-only group. None of these comparisons were statistically significant. Risk factors for acquisition of vancomycin-resistant enterococci induced length of stay in the medical intensive care unit, use of enteral feeding, and use of sucralfate. Compliance with precautions was 79% in glove-and-gown rooms and 62% in glove-only rooms (P < 0.001). Only 25 of 397 (6.3%) environmental cultures were positive for vancomycin-resistant enterococci. Nineteen types of vancomycin-resistant enterococci were documented by pulsed-field gel electrophoresis during the study period. Universal use of gloves and gowns was no better than universal use of gloves only in preventing rectal colonization by vancomycin-resistant enterococci in a medical intensive care unit of a hospital in which vancomycin-resistant enterococci are endemic. Because the use of gowns and gloves together may be associated with better compliance and may help prevent transmission of other infectious agents, this finding may not be applicable to outbreaks caused by single strains or hospitals in which the prevalence of vancomycin-resistant enterococci is low.


Deep Ordinal Regression Network for Monocular Depth Estimation.
Monocular depth estimation, which plays a crucial role in understanding 3D scene geometry, is an ill-posed problem. Recent methods have gained significant improvement by exploring image-level information and hierarchical features from deep convolutional neural networks (DCNNs). These methods model depth estimation as a regression problem and train the regression networks by minimizing mean squared error, which suffers from slow convergence and unsatisfactory local solutions. Besides, existing depth estimation networks employ repeated spatial pooling operations, resulting in undesirable low-resolution feature maps. To obtain high-resolution depth maps, skip-connections or multilayer deconvolution networks are required, which complicates network training and consumes much more computations. To eliminate or at least largely reduce these problems, we introduce a spacing-increasing discretization (SID) strategy to discretize depth and recast depth network learning as an ordinal regression problem. By training the network using an ordinary regression loss, our method achieves much higher accuracy and faster convergence in synch. Furthermore, we adopt a multi-scale network structure which avoids unnecessary spatial pooling and captures multi-scale information in parallel. The proposed deep ordinal regression network (DORN) achieves state-of-the-art results on three challenging benchmarks, i.e., KITTI [16], Make3D [49], and NYU Depth v2 [41], and outperforms existing methods by a large margin.


Aberrant Myosin 1b Expression Promotes Cell Migration and Lymph Node Metastasis of HNSCC.
Lymph node metastasis is the major clinicopathologic feature associated with poor prognosis in patients with head and neck squamous cell carcinoma (HNSCC). Here, web-based bioinformatics meta-analysis was performed to elucidate the molecular mechanism of lymph node metastasis of human HNSCC. Preferential upregulation of Myosin 1b (MYO1B) transcript in HNSCC datasets was identified. Myo1b mRNA was highly expressed in human HNSCC cells and patient tissue specimens compared with their normal counterparts as shown by quantitative PCR (qPCR) analyses. Immunohistochemistry (IHC)-detected Myo1b expression was significantly correlated with lymph node metastases in patients with oral cancer of the tongue. HNSCC with high expression of Myo1b and chemokine receptor 4 (CCR4), another metastasis-associated molecule, was strongly associated with lymph node metastasis. RNA interference (RNAi) of Myo1b in HNSCC cells, SAS and HSC4, significantly inhibited migratory and invasive abilities through decreased large protrusion formation of cell membranes. Finally, Myo1b knockdown in SAS cells significantly inhibited in vivo cervical lymph node metastases in a cervical lymph node metastatic mouse model system. Myo1b is functionally involved in lymph node metastasis of human HNSCC through enhanced cancer cell motility and is an attractive target for new diagnostic and therapeutic strategies for patients with HNSCC.