High Power Density Tower-like Triboelectric Nanogenerator for Harvesting Arbitrary Directional Water Wave Energy.
Wave energy is one of the most available energy sources in oceans. In this work, a design of high power density triboelectric nanogenerator (TENG) based on a tower structure is proposed for harvesting wave energy from arbitrary directions. Such tower-like TENG (T-TENG) consists of multiple units made of polytetrafluoroethylene balls and three-dimensional printed arc surface coated with melt adhesive reticulation nylon film. The power generation model coupled with the kinetic model for the T-TENG is proposed and discussed. The T-TENG can effectively convert arbitrary directional wave energy into electrical energy by utilizing charged balls rolling on an optimized arc surface due to ocean wave excitation. In addition, it is found that the power density of the present T-TENG increases linearly from 1.03 W/m3 to 10.6 W/m3 by increasing the units from 1 to 10 in one block. This supports that the power density of the T-TENG increases proportionally with the number of units connected in parallel without rectifiers due to its distinctive mechanism and structure. Therefore, the design of T-TENG provides an innovative and effective approach toward large-scale blue energy harvesting by connecting more blocks to form T-TENG networks.


Statistical errors and their effect on electrodiagnostic medicine.
Failure to demonstrate abnormalities on diagnostic tests in patients felt certain to have a disease is all too common, especially when symptoms are mild to moderate. One strategy used in such cases is to perform multiple tests hoping that one will confirm the clinical suspicion. An example of this strategy is found in carpal tunnel syndrome where multiple tests are often used to confirm the diagnosis. Unfortunately, this approach is often erroneous. Since no diagnostic test can perfectly discriminate between normal and abnormal, critical values are set at levels where most subjects are correctly classified. Using this methodology, approximately 2.5% of patients who are normal will be mistakenly called abnormal. This type I error, is additive when additional independent tests are performed. Since no two tests for a single condition are completely independent, the total error is less than the sum of the individual errors for each test. Even after accounting for interdependency, the total error of combined tests may be unacceptably high. If a single, highly discriminating, test is not available and multiple tests are used, abnormalities in more than a single test are needed to distinguish between normals and abnormals.


The path from the RNA world.
We describe a sequential (step by step) Darwinian model for the evolution of life from the late stages of the RNA world through to the emergence of eukaryotes and prokaryotes. The starting point is our model, derived from current RNA activity, of the RNA world just prior to the advent of genetically-encoded protein synthesis. By focusing on the function of the protoribosome we develop a plausible model for the evolution of a protein-synthesizing ribosome from a high-fidelity RNA polymerase that incorporated triplets of oligonucleotides. With the standard assumption that during the evolution of enzymatic activity, catalysis is transferred from RNA --> RNP --> protein, the first proteins in the "breakthrough organism" (the first to have encoded protein synthesis) would be nonspecific chaperone-like proteins rather than catalytic. Moreover, because some RNA molecules that pre-date protein synthesis under this model now occur as introns in some of the very earliest proteins, the model predicts these particular introns are older than the exons surrounding them, the "introns-first" theory. Many features of the model for the genome organization in the final RNA world ribo-organism are more prevalent in the eukaryotic genome and we suggest that the prokaryotic genome organization (a single, circular genome with one center of replication) was derived from a "eukaryotic-like" genome organization (a fragmented linear genome with multiple centers of replication). The steps from the proposed ribo-organism RNA genome --> eukaryotic-like DNA genome --> prokaryotic-like DNA genome are all relatively straightforward, whereas the transition prokaryotic-like genome --> eukaryotic-like genome appears impossible under a Darwinian mechanism of evolution, given the assumption of the transition RNA --> RNP --> protein. A likely molecular mechanism, "plasmid transfer," is available for the origin of prokaryotic-type genomes from an eukaryotic-like architecture. Under this model prokaryotes are considered specialized and derived with reduced dependence on ssRNA biochemistry. A functional explanation is that prokaryote ancestors underwent selection for thermophily (high temperature) and/or for rapid reproduction (r selection) at least once in their history.


Asthma and respiratory problems--a review.
Occupational asthma is the principal respiratory health problem within the primary aluminium industry. Current evidence indicates that it is irritant induced and due to occupational exposure to the inhalation of gaseous or particulate fluoride compounds. Following transfer from the occupational exposure of those who develop asthma, there is commonly symptomatic improvement. A programme of compulsory respiratory protection, progressive engineering improvements and of regular screening of potroom workers aimed at early detection, and the transfer of asthmatic workers from that environment has resulted not only in improvement of asthmatic symptoms among them, but also in the majority of an improvement in bronchial responsiveness as assessed by methacholine inhalation. The majority of studies indicate a slightly increased prevalence of symptoms of chronic bronchitis and of chronic obstructive pulmonary disease among workers in carbon bake areas, although tobacco smoking has a greater and additive effect. Only a trivial number of clinical cases of pulmonary fibrosis ascribed to aluminium compounds has been reported. Particle size limits smelter grade primary alumina reaching the alveoli of the lung.


Distribution and determinants of plasma homocysteine levels in rural Chinese twins across the lifespan.
Plasma homocysteine (Hcy) is a modifiable, independent risk factor for cardiovascular disease (CVD) and is affected by both environmental and genetic factors. This study aimed to describe the gender- and age-specific distribution of Hcy concentration for 1117 subjects aged 10-66 years, a subset of a community-based rural Chinese twin cohort. In addition, we examined environmental and genetic contributions to variances in Hcy concentration by gender and age groups. We found that the distribution pattern for Hcy varied by both age and gender. Males had higher Hcy than females across all ages. Elevated Hcy was found in 43% of male adults and 13% of female adults. Moreover, nearly one fifth of children had elevated Hcy. Genetic factors could explain 52%, 36% and 69% of the variation in Hcy concentration among children, male adults and female adults, respectively. The MTHFR C677T variant was significantly associated with Hcy concentrations. Smokers with the TT genotype had the highest Hcy levels. Overall, our results indicate that elevated Hcy is prevalent in the children and adults in this rural Chinese population. The early identification of elevated Hcy will offer a window of opportunity for the primary prevention of CVD and metabolic syndrome.


Survival of children in Cape Town known to be vertically infected with HIV-1.
To determine the survival patterns of children in Cape Town known to be vertically infected with HIV. Retrospective record review of children diagnosed with symptomatic HIV infection during the period 1 December 1990-31 May 1995. Hospitals in the Cape Town metropolitan area. 193 children were known to be vertically HIV-infected. HIV diagnosis was based on the following criteria: two positive HIV enzyme-linked immunosorbent assays (ELISAs) in children older than 15 months and a positive ELISA together with a positive polymerase chain reaction (PCR) in younger children. The mothers of the children were known to be HIV-positive. On the basis of the presenting clinical findings children were assigned to a disease severity category (A, B or C) according to the Centers for Disease Control and Prevention (CDC)'s 1994 revised classification system for HIV infection in children. Survival was analysed according to the Kaplan-Meier method. Survival time was defined as the length of time between clinical diagnosis of HIV and death or last contact with the health services. Mortality risk in relation to specific variables at diagnosis such as age and clinical manifestations was determined by calculation of odds ratios (ORs) with 95% confidence intervals (CIs). The median age at diagnosis was 5 months; 72% of children were aged less than 1 year at diagnosis. According to the CDC clinical classification, 47 (24%) fell into category A, 111 (58%) into category B and 35 (18%) into category C. Of the 193 patients 85 (44%) were alive at the time of review, 65 (34%) had died and 43 (22%) were lost to follow-up. Risk of death was significantly associated with age less than 6 months (OR 4.7, CI 2.1-10.3) and severe disease, i.e. CDC category C (OR 2.7; CI 1.1-6.9) at time of diagnosis. The median survival for all the children from time of diagnosis was 32 months. Infants diagnosed before 6 months of age had significantly shorter median survival (10 months) compared with 36 months for those diagnosed at 7-12 months of age. For the children over the age of 12 months the cumulative proportion surviving at 48 months was 78%. Children with severe disease (category C) had a median survival of 21 months, significantly lower than that in category B (32 months). For the children in category A the cumulative proportion surviving at 48 months was 66%. The median survival of children with HIV was 32 months from time of diagnosis, and survival was influenced by age and disease severity.


Motion layer extraction in the presence of occlusion using graph cuts.
Extracting layers from video is very important for video representation, analysis, compression, and synthesis. Assuming that a scene can be approximately described by multiple planar regions, this paper describes a robust and novel approach to automatically extract a set of affine or projective transformations induced by these regions, detect the occlusion pixels over multiple consecutive frames, and segment the scene into several motion layers. First, after determining a number of seed regions using correspondences in two frames, we expand the seed regions and reject the outliers employing the graph cuts method integrated with level set representation. Next, these initial regions are merged into several initial layers according to the motion similarity. Third, an occlusion order constraint on multiple frames is explored, which enforces that the occlusion area increases with the temporal order in a short period and effectively maintains segmentation consistency over multiple consecutive frames. Then, the correct layer segmentation is obtained by using a graph cuts algorithm and the occlusions between the overlapping layers are explicitly determined. Several experimental results are demonstrated to show that our approach is effective and robust.


Analytical validation of a prognostic prostate cancer gene expression assay using formalin fixed paraffin embedded tissue.
There is a clear need for assays that can predict the risk of metastatic prostate cancer following curative procedures. Importantly these assays must be analytically robust in order to provide quality data for important clinical decisions. DNA microarray based gene expression assays measure several analytes simultaneously and can present specific challenges to analytical validation. This study describes the analytical validation of one such assay designed to predict metastatic recurrence in prostate cancer using primary formalin fixed paraffin embedded tumour material. Accuracy was evaluated with a method comparison study between the assay development platform (Prostate Disease Specific Array) and an alternative platform (Xcel™ microarray) using 50 formalin-fixed, paraffin-embedded prostate cancer patient samples. An additional 70 samples were used to establish the assay reportable range. Determination of assay precision and sensitivity was performed on multiple technical replicates of three prostate cancer samples across multiple variables (operators, days, runs, reagent lots, and equipment) and RNA/cDNA inputs respectively using the appropriate linear mixed model. The overall agreement between the development and alternative platform was 94.7% (95% confidence interval, 86.9-98.5%). The reportable range was determined to be 0.150 to 1.107 for core needle biopsy samples and - 0.214 to 0.844 for radical prostatectomy samples. From the precision study, the standard deviations for assay repeatability and reproducibility were 0.032 and 0.040 respectively. The sensitivity study demonstrated that a total RNA input and cDNA input of 50 ng and 3.5 μg respectively was conservative. The Metastatic Assay was found to be highly reproducible and precise. In conclusion the studies demonstrated an acceptable analytical performance for the assay and support its potential use in the clinic.


POU domain transcription factor brain 4 confers pancreatic alpha-cell-specific expression of the proglucagon gene through interaction with a novel proximal promoter G1 element.
The proglucagon gene is expressed in a highly restricted tissue-specific manner in the alpha cells of the pancreatic islet, the hypothalamus, and the small and large intestines. Proglucagon is processed to glucagon and glucagon-like peptides GLP-1 and -2. Glucagon is expressed in alpha cells and regulates glucose homeostasis. GLP-1 is implicated in the control of insulin secretion, food intake, and satiety signaling, and GLP-2 is implicated in regulating small-bowel growth. Cell-specific expression of the proglucagon gene is mediated by proteins that interact with the proximal G1 promoter element which contains several AT-rich domains with binding sites for homeodomain transcription factors. In an attempt to identify major homeodomain proteins involved in pancreatic alpha-cell-specific proglucagon expression, we found that the POU domain transcription factor brain 4 is abundantly expressed in proglucagon-producing islet cell lines and rat pancreatic islets. In the latter, brain 4 and glucagon immunoreactivity colocalize in the outer mantle of islets. Electrophoretic mobility shift assays with specific antisera identify brain 4 as a major constituent of nuclear proteins of glucagon-producing cells that bind to the G1 element of the proglucagon gene proximal promoter. Transcriptional transactivation experiments reveal that brain 4 is a major regulator of proglucagon gene expression by its interaction with the G1 element. The finding that a neuronal transcription factor is involved in glucagon gene transcription may explain the presence of proglucagon in certain areas of the brain as well as in pancreatic alpha cells. Further, this finding supports the idea that the neuronal properties of endodermis-derived endocrine pancreatic cells may find their basis in regulation of gene expression by neuronal transcription factors.


More questions than answers.
Weight loss and obesity have been challenging topics in science and public health for decades. While scientists have pursued understanding through studies of metabolism and the microbiome, public health scholars have shown that social factors such as poverty and inadequate health care are also major contributors. And though it is true that excess weight carries with it numerous comorbidities that lower life expectancy and add costs to the health care system, studies also show that it is possible to be overweight and healthy. That doesn't change the fact that obesity can be associated with poor mental health, in part because of the corrosive societal stigma that associates being overweight with mental weakness, not biochemistry.


Risk of acute liver injury associated with the use of moxifloxacin and other oral antimicrobials: a retrospective, population-based cohort study.
To estimate the incidence and relative risk of a hospitalization or emergency visit for noninfectious liver injury in users of eight oral antimicrobials-amoxicillin, amoxicillin-clavulanic acid, clarithromycin, cefuroxime, doxycycline, levofloxacin, moxifloxacin, telithromycin-compared with nonusers of these antimicrobials. Retrospective, observational cohort study with a nested case-control analysis. HealthCore Integrated Research Database. Adults with continuous health plan enrollment for at least 6 months before study entry who had a new dispensing of a study antimicrobial between July 1, 2001, and March 31, 2009. Cases had diagnoses indicating noninfectious liver injury during follow-up. To control for potentially confounding risk factors, 10 controls at risk for liver injury during follow-up were matched to each case by age, sex, and event date (liver injury date of the case), and analyses were adjusted for medical history, concomitant drugs, and health care service use. Two physician reviewers (blind to exposure) validated the cases. Among 1.3 million antimicrobial users, we identified 607 cases of liver injury, including 82 cases of severe hepatocellular injury and 11 cases of liver failure. Liver injury incidence in nonusers of study antimicrobials was 35/100,000 person-years (95% confidence interval [CI] 29-42/100,000 person-years). For valid cases, the adjusted relative risk among current users of multiple antimicrobials was 3.2 (95% CI 1.6-6.7). Levofloxacin had the highest relative risk for current single use (3.2, 95% CI 1.8-5.8). Relative risks were also elevated for amoxicillin-clavulanic acid (2.5, 95% CI 1.3-5.0), doxycycline (2.5, 95% CI 1.2-5.2), moxifloxacin (2.3, 95% CI 1.1-4.7), and amoxicillin (2.3, 95% CI 1.1-4.7). The results support a comparatively high adjusted relative risk of liver injury among patients exposed concurrently to multiple antimicrobials and modest elevations in the risk for several antimicrobials used alone; however, we found little evidence of any strong effect of commonly used antimicrobials on the risk of liver injury.


The X-ray crystal structure of RNA polymerase from Archaea.
The transcription apparatus in Archaea can be described as a simplified version of its eukaryotic RNA polymerase (RNAP) II counterpart, comprising an RNAPII-like enzyme as well as two general transcription factors, the TATA-binding protein (TBP) and the eukaryotic TFIIB orthologue TFB. It has been widely understood that precise comparisons of cellular RNAP crystal structures could reveal structural elements common to all enzymes and that these insights would be useful in analysing components of each enzyme that enable it to perform domain-specific gene expression. However, the structure of archaeal RNAP has been limited to individual subunits. Here we report the first crystal structure of the archaeal RNAP from Sulfolobus solfataricus at 3.4 A resolution, completing the suite of multi-subunit RNAP structures from all three domains of life. We also report the high-resolution (at 1.76 A) crystal structure of the D/L subcomplex of archaeal RNAP and provide the first experimental evidence of any RNAP possessing an iron-sulphur (Fe-S) cluster, which may play a structural role in a key subunit of RNAP assembly. The striking structural similarity between archaeal RNAP and eukaryotic RNAPII highlights the simpler archaeal RNAP as an ideal model system for dissecting the molecular basis of eukaryotic transcription.


Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretory baculovirus system.
In a step towards a tetravalent dengue virus subunit vaccine which is economical to produce, highly immunogenic and stable, a hybrid dengue virus envelope (E) protein molecule has been constructed. It consists of 36 amino acids from the membrane protein, the N-terminal 288 amino acids of the dengue-2 virus E protein plus amino acids 289-424 of the dengue-3 virus E protein. It has been engineered for secretory expression by fusion to a mellitin secretory signal sequence and truncation of the hydrophobic transmembrane segment. Using the baculovirus expression system and serum-free conditions, more than 95% of recombinant dengue-2 virus-dengue-3 virus hybrid E protein (rD2D3E) was secreted into the cell culture supernatant in a stable form with multiple features indicative of preserved conformation. The hybrid molecule reacted with a panel of dengue virus- and flavivirus-specific MAbs which recognize linear or conformational epitopes on dengue virions. Human dengue virus-specific antisera also reacted with the protein. The hybrid rD2D3E protein was able to inhibit the in vitro binding of dengue-2 and dengue-3 viruses to human myelomonocytic cells, suggesting that the receptor-binding epitope(s) was preserved. Adjuvant-free immunization with the hybrid protein induced an antibody response to both dengue-2 and dengue-3 virus in outbred mice, comparable in strength to that of individual rD2E and rD3E proteins. Notably, these antibody responses were primarily of the IgG2a and IgG2b isotype. A strong dengue virus cross-reactive T cell response was also induced in the mice, suggesting that dengue virus hybrid E proteins could form the basis of an efficacious multivalent dengue virus vaccine.


Brain volumetric correlates of autism spectrum disorder symptoms in attention deficit/hyperactivity disorder.
Autism spectrum disorder (ASD) symptoms frequently occur in subjects with attention deficit/hyperactivity disorder (ADHD). While there is evidence that both ADHD and ASD have differential structural correlates, no study to date has investigated these structural correlates within a framework that robustly accounts for the phenotypic overlap between the two disorders. The presence of ASD symptoms was measured by the parent-reported Children's Social and Behavioural Questionnaire (CSBQ) in ADHD subjects (n = 180), their unaffected siblings (n = 118) and healthy controls (n = 146). ADHD symptoms were assessed by a structured interview (K-SADS-PL) and the Conners' ADHD questionnaires. Whole brain T1-weighted MPRAGE images were acquired and the structural MRI correlates of ASD symptom scores were analysed by modelling ASD symptom scores against white matter (WM) and grey matter (GM) volumes using mixed effects models which controlled for ADHD symptom levels. ASD symptoms were significantly elevated in ADHD subjects relative to both controls and unaffected siblings. ASD scores were predicted by the interaction between WM and GM volumes. Increasing ASD score was associated with greater GM volume. Equivocal results from previous structural studies in ADHD and ASD may be due to the fact that comorbidity has not been taken into account in studies to date. The current findings stress the need to account for issues of ASD comorbidity in ADHD.


A Survey on Trust Management for WBAN: Investigations and Future Directions.
The rapid uptake of the Wireless Body Area Networks (WBAN) and their services poses unprecedented security requirements. WBAN are evolving to support these requirements. Fulfilling these tasks is challenging as their mobile context is increasingly complex, heterogeneous, and evolving. One potential solution to meet the WBAN security requirements is trust management that helps to reach a more secure and adaptable WBAN environment. Accordingly, this article aims to serve as a brief survey of trust management approaches within intra-WBAN and inter-WBAN. For that, we first summarize trust management concepts, discuss WBAN challenges and classify the attacks on WBAN trust management models. Subsequently, we detail and compare the existing trust based approaches in a WBAN context. We pinpoint their limitations and provide a new classification of these different approaches. We also propose a set of best practices that may help the reader to build a robust and an efficient trust management framework. We complete this survey by highlighting the open future directions and perspectives for research.


MP3: a software tool for the prediction of pathogenic proteins in genomic and metagenomic data.
The identification of virulent proteins in any de-novo sequenced genome is useful in estimating its pathogenic ability and understanding the mechanism of pathogenesis. Similarly, the identification of such proteins could be valuable in comparing the metagenome of healthy and diseased individuals and estimating the proportion of pathogenic species. However, the common challenge in both the above tasks is the identification of virulent proteins since a significant proportion of genomic and metagenomic proteins are novel and yet unannotated. The currently available tools which carry out the identification of virulent proteins provide limited accuracy and cannot be used on large datasets. Therefore, we have developed an MP3 standalone tool and web server for the prediction of pathogenic proteins in both genomic and metagenomic datasets. MP3 is developed using an integrated Support Vector Machine (SVM) and Hidden Markov Model (HMM) approach to carry out highly fast, sensitive and accurate prediction of pathogenic proteins. It displayed Sensitivity, Specificity, MCC and accuracy values of 92%, 100%, 0.92 and 96%, respectively, on blind dataset constructed using complete proteins. On the two metagenomic blind datasets (Blind A: 51-100 amino acids and Blind B: 30-50 amino acids), it displayed Sensitivity, Specificity, MCC and accuracy values of 82.39%, 97.86%, 0.80 and 89.32% for Blind A and 71.60%, 94.48%, 0.67 and 81.86% for Blind B, respectively. In addition, the performance of MP3 was validated on selected bacterial genomic and real metagenomic datasets. To our knowledge, MP3 is the only program that specializes in fast and accurate identification of partial pathogenic proteins predicted from short (100-150 bp) metagenomic reads and also performs exceptionally well on complete protein sequences. MP3 is publicly available at http://metagenomics.iiserb.ac.in/mp3/index.php.


Tailoring relaxation time spectrum in soft glassy materials.
Physical properties of out of equilibrium soft materials depend on time as well as deformation history. In this work we propose to transform this major shortcoming into gain by applying controlled deformation field to tailor the rheological properties. We take advantage of the fact that deformation field of a certain magnitude can prevent particles in an aging soft glassy material from occupying energy wells up to a certain depth, thereby populating only the deeper wells. We employ two soft glassy materials with dissimilar microstructures and demonstrate that increase in strength of deformation field while aging leads to narrowing of spectrum of relaxation times. We believe that, in principle, this philosophy can be universally applied to different kinds of glassy materials by changing nature and strength of impetus.


Intraspecific interactions in a high-density leopard population.
Although less studied than interspecific interactions, interactions among members of the same species can influence space use and temporal activity. Using techniques commonly applied to the analysis of interspecific interactions-multispecies occupancy modeling and the analysis of temporal activity patterns-we studied intraspecific interactions within a high-density population of Persian leopards (Panthera pardus saxicolor) in Tandoureh National Park, northeastern Iran. Using camera-trap data, we investigated spatiotemporal interactions between male leopards, lone female leopards, and families (cubs/females with cubs). While we hypothesized that male and female leopards would display different temporal activity patterns, we did not predict spatial avoidance between these groups. We also predicted that leopard families would exhibit spatiotemporal avoidance from male leopards due to the risk of infanticide. Contrary to our expectations, we did not find any evidence for spatial or temporal avoidance between leopard families and adult male leopards. Male and lone female leopards exhibited positive pairwise co-occurrence, consistent with reports of high overlap between male and female leopard home ranges. While a high level of overlap in temporal activity patterns was found between males/lone females and males/families, there was evidence for variation in the proportion of time each leopard group was active in particular periods of the diel cycle. Male leopards showed cathemeral activity, while lone females and families were more active during daylight hours. The application of these techniques to interactions within a species has improved understanding of the ecology and behavior of this endangered solitary carnivore.


Predictive model for major complications 2 years after corrective spine surgery for adult spinal deformity.
ASD surgery improves a patient's health-related quality of life, but it has a high complication rate. The aim of this study was to create a predictive model for complications after surgical treatment for adult spinal deformity (ASD), using spinal alignment, demographic data, and surgical invasiveness. This study included 195 surgically treated ASD patients who were > 50 years old and had 2-year follow-up from multicenter database. Variables which included age, gender, BMI, BMD, frailty, fusion level, UIV and LIV, primary or revision surgery, pedicle subtraction osteotomy, spinal alignment, Schwab-SRS type, surgical time, and blood loss were recorded and analyzed at least 2 years after surgery. Decision-making trees for 2-year postoperative complications were constructed and validated by a 7:3 data split for training and testing. External validation was performed for 25 ASD patients who had surgery at a different hospital. Complications developed in 48% of the training samples. Almost half of the complications developed in late post-op period, and implant-related complications were the most common complication at 2 years after surgery. Univariate analyses showed that BMD, frailty, PSO, LIV, PI-LL, and EBL were risk factors for complications. Multivariate analysis showed that low BMD, PI-LL > 30°, and frailty were independent risk factors for complications. In the testing samples, our predictive model was 92% accurate with an area under the receiver operating characteristic curve of 0.963 and 84% accurate in the external validation. A successful model was developed for predicting surgical complications. Our model could inform physicians about the risk of complications in ASD patients in the 2-year postoperative period. These slides can be retrieved under Electronic Supplementary Material.


STAT6 mediates interleukin-4 growth inhibition in human breast cancer cells.
In addition to acting as a hematopoietic growth factor, interleukin-4 (IL-4) inhibits growth of some transformed cells in vitro and in vivo. In this study, we show that insulin receptor substrate (IRS)-1, IRS-2, and signal transducer and activator of transcription 6 (STAT6) are phosphorylated following IL-4 treatment in MCF-7 breast cancer cells. STAT6 DNA binding is enhanced by IL-4 treatment. STAT6 activation occurs even after IRS-1 depletion, suggesting the two pathways are independent. To examine the role of STAT6 in IL-4-mediated growth inhibition and apoptosis, a full-length STAT6 cDNA was transfected into MCF-7 cells. Transient overexpression of STAT6 resulted in both cytoplasmic and nuclear expression of the protein, increased DNA binding in response to IL-4, and increased transactivation of an IL-4 responsive promoter. In STAT6-transfected cells, basal proliferation was reduced whereas apoptosis was increased. Finally, stable expression of STAT6 resulted in reduced foci formation compared to vector-transfected cells alone. These results suggest STAT6 is required for IL-4-mediated growth inhibition and induction of apoptosis in human breast cancer cells.


Regulation of cytochrome P-45011 beta gene expression by adrenocorticotropin.
Recombinant DNA clones corresponding to 11 beta-hydroxylase cytochrome P-450 (P-450(11)beta) have been identified in a bovine adrenocortical cDNA library. These clones, pB11 beta-1 and pB11 beta-2, hybridize to at least three RNA species, 7.2, 6.2, and 4.3 kilobases in length, present in the adrenal cortex. All three RNA species directed cytochrome P-450(11)beta synthesis in an in vitro translation system. Expression of the cytochrome P-450(11)beta gene is tissue specific in that these transcripts were not detected in liver, heart, kidney, or corpus luteum. In cultured bovine adrenocortical cells, adrenocorticotropin (ACTH) or dibutyryl cAMP increased the concentration of cytochrome P-450(11)beta transcripts. This increase appears to be at least partially due to de novo transcription as shown by the action of actinomycin D which effectively blocked the ACTH-induced increase in cytochrome P-450(11)beta RNA level. Furthermore, cycloheximide administered prior to or along with ACTH resulted in the blockage of any new transcription of the cytochrome P-450(11)beta gene as evidenced from the level of RNA. Thus, in addition to potential effects on RNA stabilization, a primary action of ACTH in the regulation of the level of cytochrome P-450(11)beta may be to mediate the induction of an unidentified adrenocortical protein which is required for the activation of cytochrome P-450(11)beta gene expression. These results, together with previous studies (Kramer, R.E., Rainey, W.E., Funkenstein, B., Dee, A., Simpson, E. R., and Waterman, M. R. (1984) J. Biol. Chem. 259, 707-713) that showed increased intracellular levels of cAMP upon treatment of bovine adrenocortical cell cultures with ACTH, suggest a role for cAMP-mediated events in the regulation of cytochrome P-450(11)beta gene expression.


The nutritional significance of lipid rafts.
The structure, size, stability, and functionality of lipid rafts are still in debate, but recent techniques allowing direct visualization have characterized them in a wide range of cell types. Lipid rafts are potentially modifiable by diet, particularly (but not exclusively) by dietary fatty acids. However, it is not clear whether dietary polyunsaturated fatty acids (PUFAs) are incorporated into raft lipids or whether their low affinity to cholesterol disallows this and causes phase separation from rafts and displacement of raft proteins. This review examines the potential for dietary modification of raft structure and function in the immune system, brain and retinal tissue, the gut, and in cancer cells. Although there is increasing evidence to suggest that membrane microdomains, and their modulation, have an impact in health and disease, it is too early to judge whether modulation of lipid rafts is responsible for the immunomodulatory effects of n-3 PUFA. In addition to dietary fatty acids, gangliosides and cholesterol may also modulate microdomains in a number of tissues, and recent work has highlighted sphingolipids in membrane microdomains as potential targets for inhibition of tumor growth by n-3 PUFA. The roles of fatty acids and gangliosides in cognitive development, age-related cognitive decline, psychiatric disorders, and Alzheimer's disease are poorly understood and require clarification, particularly with respect to the contribution of lipid rafts. The roles of lipid rafts in cancer, in microbial pathogenesis, and in insulin resistance are only just emerging, but compelling evidence indicates the growing importance of membrane microdomains in health and disease.


Resection and repair of a Cas9 double-strand break at CTG trinucleotide repeats induces local and extensive chromosomal deletions.
Microsatellites are short tandem repeats, ubiquitous in all eukaryotes and represent ~2% of the human genome. Among them, trinucleotide repeats are responsible for more than two dozen neurological and developmental disorders. Targeting microsatellites with dedicated DNA endonucleases could become a viable option for patients affected with dramatic neurodegenerative disorders. Here, we used the Streptococcus pyogenes Cas9 to induce a double-strand break within the expanded CTG repeat involved in myotonic dystrophy type 1, integrated in a yeast chromosome. Repair of this double-strand break generated unexpected large chromosomal deletions around the repeat tract. These deletions depended on RAD50, RAD52, DNL4 and SAE2, and both non-homologous end-joining and single-strand annealing pathways were involved. Resection and repair of the double-strand break (DSB) were totally abolished in a rad50Δ strain, whereas they were impaired in a sae2Δ mutant, only on the DSB end containing most of the repeat tract. This observation demonstrates that Sae2 plays significant different roles in resecting a DSB end containing a repeated and structured sequence as compared to a non-repeated DSB end. In addition, we also discovered that gene conversion was less efficient when the DSB could be repaired using a homologous template, suggesting that the trinucleotide repeat may interfere with gene conversion too. Altogether, these data show that SpCas9 may not be the best choice when inducing a double-strand break at or near a microsatellite, especially in mammalian genomes that contain many more dispersed repeated elements than the yeast genome.


Molecular Epidemiology and Mechanism of Sulbactam Resistance in Acinetobacter baumannii Isolates with Diverse Genetic Backgrounds in China.
Sulbactam is a plausible option for treating Acinetobacter infections because of its intrinsic antibacterial activity against the members of the Acinetobacter genus, but the mechanisms of sulbactam resistance have not been fully studied in Acinetobacter baumannii In this study, a total of 2,197 clinical A. baumannii isolates were collected from 27 provinces in China. Eighty-eight isolates with various MICs for sulbactam were selected on the basis of their diverse clonality and underwent multilocus sequence typing (MLST), antimicrobial susceptibility testing, and resistance gene screening. The copy number and relative expression of blaTEM-1D and ampC were measured via quantitative PCR and quantitative reverse transcription-PCR, respectively. The genetic structure of multicopy blaTEM-1D was determined using the whole-genome sequencing technology. The cefoperazone-sulbactam resistance rate of the 2,197 isolates was 39.7%. The rate of positivity for blaTEM-1D or ISAba1-ampC in the sulbactam-nonsusceptible group (64.91% and 78.95%, respectively) was significantly higher than that in the sulbactam-susceptible group (0% and 0%, respectively; P < 0.001). The MIC of sulbactam (P < 0.001) varied considerably between the groups expressing ampC with or without upstream ISAba1 Notably, the genetic structure of the multicopy blaTEM-1D gene in strain ZS3 revealed that blaTEM-1D was embedded within four tandem copies of the cassette IS26-blaTEM-1D-Tn3-IS26 Therefore, blaTEM-1D and ISAba1-ampC represent the prevalent mechanism underlying sulbactam resistance in clinical A. baumannii isolates in China. The structure of the four tandem copies of blaTEM-1D first identified may increase sulbactam resistance.


Evasion of the alternative complement pathway by metacyclic trypomastigotes of Trypanosoma cruzi: dependence on the developmentally regulated synthesis of surface protein and N-linked carbohydrate.
Epimastigotes (EPI) of Trypanosoma cruzi are highly sensitive to lysis in fresh normal human serum by the alternative complement pathway (ACP). In contrast, metacyclic trypomastigotes (CMT) derived from EPI in stationary culture fail to activate the ACP and are thus resistant to serum-mediated lysis. To investigate the nature of the parasitic surface molecules which enable infective metacyclic trypomastigotes to evade the ACP, CMT were treated with a variety of different proteolytic and glycosidic enzymes, and their sensitivity to ACP-dependent lysis was tested. Pretreatment with pronase was found to cause a near complete reversal in the resistance of CMT to serum lysis, whereas trypsin or chymotrypsin induced smaller increases in complement sensitivity. Similarly, pretreatment with N-glycanase or neuraminidase also partially abrogated the resistance of CMT to ACP-dependent lysis. The effect of these enzymes on susceptibility to complement-mediated lysis was paralleled in increased C3 and C9 deposition on the organism. In addition, electrophoretic analysis of parasite-bound C3 indicated that the hemolytically inactive fragment, iC3b, was the major form of the molecule on CMT, while the hemolytically active fragment, C3b, predominated on pronase-treated CMT. Furthermore, when C3 was deposited on the parasite surface by means of purified ACP components, 80% of C3b on pronase-pretreated CMT but only 14% of the C3b on CMT bound the amplification protein factor B with high affinity, a prerequisite for efficient ACP activation. When cultured at 37 degrees C after pronase treatment, CMT gradually regained their resistance to ACP-mediated lysis. This process was blocked if puromycin, cycloheximide, or tunicamycin were included in the culture medium. The above findings suggest that evasion of the ACP by CMT is dependent on the developmentally regulated synthesis of protein as well as N-linked carbohydrate chains. A stage-specific 90,000 to 115,000 m.w. glycoprotein doublet present on the surface of CMT was shown to be uniquely sensitive to pronase digestion. Thus, this complex, which is also recognized by a CMT-specific monoclonal antibody, may be the glycoprotein component responsible for control of ACP activation


"They'll Always Find a Way to Get to You": Technology Use in Adolescent Romantic Relationships and Its Role in Dating Violence and Abuse.
Electronic communication technology (ECT), such as mobile phones and online communication tools, is widely used by adolescents; however, the availability of such tools may have both positive and negative impacts within the context of romantic relationships. While an established literature has documented the nature, prevalence, and impact of traditional forms of adolescent dating violence and abuse (ADVA), limited empirical investigation has focused on the role of ECT in ADVA or what shall be termed technology-assisted adolescent dating violence and abuse (TAADVA) and how adolescents perceive the impact of TAADVA relative to ADVA. In this article, the authors explore the role ECT plays in adolescent romantic relationships and psychologically abusive and controlling ADVA behaviors and its perceived impact. An opportunity sample of 52 adolescents (22 males and 30 females) between the ages of 12 and 18 years participated in the study. One all-female and seven mixed-gendered semi-structured focus groups were conducted. Thematic analysis was used to identify three superordinate themes, including (a) perceived healthy versus unhealthy communication, (b) perceived monitoring and controlling communication, and (c) perceived impact of technology-assisted abuse compared with that in person. While ECTs had a positive impact on the development and maintenance of adolescent romantic relationships, such tools also provided a new avenue for unhealthy, harassment, monitoring, and controlling behaviors within these relationships. ECT was also perceived to provide unique impacts in terms of making TAADVA seem both less harmful and more harmful than ADVA experienced in person. Adolescents' perceptions and experiences of ECT in romantic relationships and TAADVA may also vary be gender. Implications of the findings are discussed, and recommendations are made for future research.


Quantitative assay for epinephrine in dental anesthetic solutions by capillary electrophoresis.
A simple and robust method for the separation and quantification of epinephrine in dental anesthetic solutions was developed. The method allows the direct injection of high salt solutions without sample pre-treatment. Large sample plugs (5.7% of the total capillary length) are used for epinephrine determination by selective analyte focusing in capillary electrophoresis. The concentration detection limit for epinephrine is about 5.0 x 10(-7) M (90 ng ml-1) with a commercial UV detector. The separation protocol was validated in terms of its precision, linearity, accuracy and specificity.


Beyond Comorbidity: A Critical Perspective of Syndemic Depression and Diabetes in Cross-cultural Contexts.
This article examines the comorbidity concept in medical anthropology. I argue that the dearth of articles on comorbidity in medical anthropology may result from the rise of syndemic theory. Syndemics recognize how social realities shape individual illness experiences as well as distribution of diseases across populations. I discuss synergistic interactions foundational to the syndemics construct through my research of depression and diabetes comorbidity in vulnerable populations from urban United States, India, and South Africa. I argue that social and economic factors that cluster with depression and diabetes alone and together exemplify the biosocial processes that are at the heart of syndemics. In doing so, I illustrate how social, cultural, and economic factors shape individual-level experiences of co-occurring diseases despite similar population-level trends. Finally, I discuss the relevance of syndemics for the fields of medicine and public health while cautioning what must not be lost in translation across disciplines.


About Politeness, Face, and Feedback: Exploring Resident and Faculty Perceptions of How Institutional Feedback Culture Influences Feedback Practices.
To explore resident and faculty perspectives on what constitutes feedback culture, their perceptions of how institutional feedback culture (including politeness concepts) might influence the quality and impact of feedback, feedback seeking, receptivity, and readiness to engage in bidirectional feedback. Using a constructivist grounded theory approach, five focus group discussions with internal medicine residents, three focus group discussions with general medicine faculty, and eight individual interviews with subspecialist faculty were conducted at Brigham and Women's Hospital between April and December 2016. Discussions and interviews were audiotaped and transcribed verbatim; concurrent data collection and analysis were performed using the constant comparative approach. Analysis was considered through the lens of politeness theory and organizational culture. Twenty-nine residents and twenty-two general medicine faculty participated in focus group discussions, and eight subspecialty faculty participated in interviews. The institutional feedback culture was described by participants as (1) a culture of politeness, in which language potentially damaging to residents' self-esteem was discouraged; and (2) a culture of excellence, in which the institution's outstanding reputation and pedigree of trainees inhibited constructive feedback. Three key themes situated within this broader cultural context were discovered: normalizing constructive feedback to promote a culture of growth, overcoming the mental block to feedback seeking, and hierarchical culture impeding bidirectional feedback. An institutional feedback culture of excellence and politeness may impede honest, meaningful feedback and may impact feedback seeking, receptivity, and bidirectional feedback exchanges. It is essential to understand the institutional feedback culture before it can be successfully changed.


What causes the experience of discrimination in non-regular workers?
Discrimination based on type of employment against non-regular workers is still a social issue. However, there are few studies on job factors that affect the discrimination experience in each type of employment or the association between discrimination and health impact indicators. This study examined occupational health characteristics according to discrimination experience and relating factors that affect discrimination experience. This study used the 4th Korean Working Conditions Survey (2014) provided by the Korea Occupational Safety and Health Agency. Among the 50,000 workers, 7731 non-regular wage workers were selected as study population. To examine differences in discrimination experience, we used a t-test on occupational risk factors, occupational stress, occupational characteristics, health impact indicators. To identify the factors that affected discrimination experience, we performed binomial logistic regression analysis. The discrimination experience rate was significantly higher in male, aged less than 40 years old, above high school graduate than middle school graduate, higher wage level, shorter employment period and larger company's scale. As factors related to discrimination experience, they experienced discrimination more as occupational stress was higher and when they were temporary or daily workers rather than permanent workers, work patterns were not consistent, and the support of boss was low. It showed that physical, musculoskeletal, and mental occupational risk scores and subjective job instability were higher and work environment satisfaction was lower in discrimination experienced group. The present study showed that the demographic and occupational factors were complexly related to discrimination experience in non-regular workers. The experience of discrimination had increased when occupational stress was higher, they were temporary or daily workers rather than permanent workers, work patterns were not consistent, and their boss' support was low. Improving various relating factors, (e.g. occupational stresses, employment status and occupational characteristics), this would ultimately expect to improve non-regular workers' discrimination.


Importance effects on performance in event-based prospective memory tasks.
In the present study we manipulated the importance of performing two event-based prospective memory tasks. In Experiment 1, the event-based task was assumed to rely on relatively automatic processes, whereas in Experiment 2 the event-based task was assumed to rely on a more demanding monitoring process. In contrast to the first experiment, the second experiment showed that importance had a positive effect on prospective memory performance. In addition, the occurrence of an importance effect on prospective memory performance seemed to be mainly due to the features of the prospective memory task itself, and not to the characteristics of the ongoing tasks that only influenced the size of the importance effect. The results suggest that importance instructions may improve prospective memory if the prospective task requires the strategic allocation of attentional monitoring resources.


Dramatic impact of using protective equipment on the level of hurling-related head injuries: an ultimately successful 27-year programme.
Major head injuries are not uncommon in the Irish national game of hurling. Historically, helmets were not worn. We report a multistage campaign to facilitate and encourage the use of appropriate headgear among the estimated 100 000 hurling players in Ireland. This campaign lasted for 27 years between 1985 and 2012, and involved a number of different stages including: (1) facilitating the establishment of a business dedicated to developing head protection equipment suitable for hurling, (2) placing a particular emphasis on continual product enhancement to the highest industrial standards, (3) engaging continually with the game's controlling body, the Gaelic Athletic Association (GAA), with the ultimate objective of securing a mandatory usage policy for protective helmets and faceguards, (4) longitudinal research to monitor hurling injury, equipment usage and players' attitudes and (5) widely communicating key research findings to GAA leaders and members, as well as to 1000 clubs and schools. One of our three relevant studies included 798 patients and identified a dramatic association between the type of head protection used by a player, if any, and the site of the injury requiring treatment. While 51% of the injured players without head protection suffered head trauma, this rate was only 35% among the players wearing helmets and 5% among players who were wearing full head protection (both a helmet and faceguard). The GAA responded in three stages to the accumulating evidence: (1) they introduced a mandatory regulation for those aged less than 18 years in 2005; (2) this ruling was extended to all players under 21 years in 2007 and (3) finally extended to all players irrespective of age, gender or grade from January 2010. The latter ruling applied to both games and organised training sessions.


Effects of acute physical exercise characteristics on cognitive performance.
The effect of physical exercise on mental function has been widely studied from the beginning of the 20th century. However, the contradictory findings of experimental research have led authors to identify several methodological factors to control in such studies including: (i) the nature of the psychological task; and (ii) the intensity and duration of physical exercise. The purpose of this article is to provide information, from the perspective of performance optimisation, on the main effects of physical task characteristics on cognitive performance. Within this framework, some consistent results have been observed during the last decade. Recent studies, using mainly complex decisional tasks, have provided the research community with clear support for an improvement of cognitive performance during exercise. Diverse contributing factors have been suggested to enhance cognitive efficacy. First, an increase in arousal level related to physical exertion has been hypothesised. Improvement in decisional performance has been observed immediately after the adrenaline threshold during incremental exercise. Such positive effects could be enhanced by nutritional factors, such as carbohydrate or fluid ingestion, but did not seem to be influenced by the level of fitness. Second, the mediating role of resource allocation has been suggested to explain improvement in cognitive performance during exercise. This effect highlights the importance of motivational factors in such tasks. Finally, when the cognitive performance was performed during exercise, consistent results have indicated that the dual task effect was strongly related to energetic constraints of the task. The greater the energy demand, the more attention is used to control movements.


Passive Double-Sensory Evoked Coherence Correlates with Long-Term Memory Capacity.
HIGHLIGHTS Memory correlates with the difference between single and double-sensory evoked steady-state coherence in the gamma range (ΔC).The correlation is most pronounced for the anterior brain region (ΔCA ).The correlation is not driven by birth size, education, speed of processing, or intelligence.The sensitivity of ΔCA for detecting low memory capacity is 90%. Cerebral rhythmic activity and oscillations are important pathways of communication between cortical cell assemblies and may be key factors in memory. We asked whether memory performance is related to gamma coherence in a non-task sensory steady-state stimulation. We investigated 40 healthy males born in 1953 who were part of a Danish birth cohort study. Coherence was measured in the gamma range in response to a single-sensory visual stimulation (36 Hz) and a double-sensory combined audiovisual stimulation (auditive: 40 Hz; visual: 36 Hz). The individual difference in coherence (ΔC) between the bimodal and monomodal stimulation was calculated for each subject and used as the main explanatory variable. ΔC in total brain were significantly negatively correlated with long-term verbal recall. This correlation was pronounced for the anterior region. In addition, the correlation between ΔC and long-term memory was robust when controlling for working memory, as well as a wide range of potentially confounding factors, including intelligence, length of education, speed of processing, visual attention and executive function. Moreover, we found that the difference in anterior coherence (ΔCA ) is a better predictor of memory than power in multivariate models. The sensitivity of ΔCA for detecting low memory capacity is 92%. Finally, ΔCA was also associated with other types of memory: verbal learning, visual recognition, and spatial memory, and these additional correlations were also robust enough to control for a range of potentially confounding factors. Thus, the ΔC is a predictor of memory performance may be useful in cognitive neuropsychological testing.


Social inequality and child malnutrition in four Andean countries.
To analyze the effects of socioeconomic, regional, and ethnic conditions on chronic malnutrition in four Andean countries of South America: Bolivia, Colombia, Ecuador, and Peru. The study was based on Demographic and Health Surveys (DHS) for Colombia (1995), Peru (1996), and Bolivia (1997), and on a Living Standard Measurement Survey for Ecuador (1998). We developed an index of household socioeconomic status using categorical principal components analysis. We broke down the prevalence of stunting by socioeconomic status (SES), ethnicity, place of residence (large cities, small cities, towns, and countryside), and region (highland region versus other areas of the country). We applied smoothed regression curves and linear functions to analyze SES effects on stunting, with specific models for Bolivia, Ecuador, and Peru. Bolivia, Ecuador, and Peru have similar characteristics, with high stunting prevalences overall; higher stunting prevalences in their highland areas, particularly among indigenous populations; and strong socioeconomic disparities. Colombia, in contrast, has a lower stunting prevalence and smaller regional disparities. The socioeconomic gradient of stunting is strong in all four countries, with prevalence rates in the poorest deciles at least three times as high as those in the top decile. The sharp contrast between the conditions found in Bolivia, Ecuador, and Peru and those in Colombia may be the result of specific ethnic factors affecting indigenous groups; a particular diet profile in the highland areas, with low protein and micronutrient intake; and differences in the long-term economic and social development paths that the countries have taken. Along with the strong socioeconomic gradient in all the countries, the weight of ethnic and regional factors suggests the need to reduce inequality as well as to comprehensively improve education and housing, better target health and nutrition programs, and implement participatory programs integrated into indigenous cultures.


The phenolic compounds of olive oil: structure, biological activity and beneficial effects on human health.
The Mediterranean diet is rich in vegetables, cereals, fruit, fish, milk, wine and olive oil and has salutary biological functions. Epidemiological studies have shown a lower incidence of atherosclerosis, cardiovascular diseases and certain kinds of cancer in the Mediterranean area. Olive oil is the main source of fat, and the Mediterranean diet's healthy effects can in particular be attributed not only to the high relationship between unsaturated and saturated fatty acids in olive oil but also to the antioxidant property of its phenolic compounds. The main phenolic compounds, hydroxytyrosol and oleuropein, which give extra-virgin olive oil its bitter, pungent taste, have powerful antioxidant activity both in vivo and in vitro. The present review focuses on recent works analysing the relationship between the structure of olive oil polyphenolic compounds and their antioxidant activity. These compounds' possible beneficial effects are due to their antioxidant activity, which is related to the development of atherosclerosis and cancer, and to anti-inflammatory and antimicrobial activity.


The role of microglia in processing and spreading of bioactive tau seeds in Alzheimer's disease.
Misfolding of microtubule-associated protein tau (MAPT) within neurons into neurofibrillary tangles is an important pathological feature of Alzheimer's disease (AD). Tau pathology correlates with cognitive decline in AD and follows a stereotypical anatomical course; several recent studies indicate that tau pathology spreads inter-neuronally via misfolded tau "seeds." Previous research has focused on neurons as the source of these tau seeds. However, recent studies as well as the data contained herein suggest that microglia, the resident immune cells of the central nervous system, play a direct role in the spread of tau pathology. Primary adult microglia were isolated from human AD cases and the rTg4510 tauopathy mouse model and used for analysis of gene expression, tau protein by Simoa technology, and quantification of tau seeding using a highly sensitive fluorescence resonance energy transfer (FRET) biosensing cell line for tau seeding and aggregation. Here, we show that microglia isolated from both human tauopathy and AD cases and the rTg4510 tauopathy mouse model stably contain tau seeds, despite not synthesizing any tau. Microglia releases these tau seeds in vitro into their conditioned media (CM). This suggests that microglia have taken up tau but are incapable of entirely neutralizing its seeding activity. Indeed, when in vitro microglia are given media containing tau seeds, they reduce (but do not eliminate) tau seeding. When microglia are treated with inflammagens such as lipopolysaccharide (LPS), interleukin-1β (IL1β), tumor necrosis factor α (TNFα), or amyloid-β, their ability to reduce tau seeding is unchanged and these factors do not induce seeding activity on their own. Overall, these data suggest that microglia have a complex role: they are capable of taking up and breaking down seed competent tau, but do so inefficiently and could therefore potentially play a role in the spread of tau pathology.


The anti-myeloma activity of bone morphogenetic protein 2 predominantly relies on the induction of growth arrest and is apoptosis-independent.
Multiple myeloma (MM), a malignancy of the bone marrow, is characterized by a pathological increase in antibody-producing plasma cells and an increase in immunoglobulins (plasmacytosis). In recent years, bone morphogenetic proteins (BMPs) have been reported to be activators of apoptotic cell death in neoplastic B cells in MM. Here, we use bone morphogenetic protein 2 (BMP2) to show that the "apoptotic" effect of BMPs on human neoplastic B cells is dominated by anti-proliferative activities and cell cycle arrest and is apoptosis-independent. The anti-proliferative effect of BMP2 was analysed in the human cell lines KMS12-BM and L363 using WST-1 and a Coulter counter and was confirmed using CytoTox assays with established inhibitors of programmed cell death (zVAD-fmk and necrostatin-1). Furthermore, apoptotic activity was compared in both cell lines employing western blot analysis for caspase 3 and 8 in cells treated with BMP2 and FasL. Additionally, expression profiles of marker genes of different cell death pathways were analysed in both cell lines after stimulation with BMP2 for 48h using an RT-PCR-based array. In our experiments we observed that there was rather no reduction in absolute cell number, but cells stopped proliferating following treatment with BMP2 instead. The time frame (48-72 h) after BMP2 treatment at which a reduction in cell number is detectable is too long to indicate a directly BMP2-triggered apoptosis. Moreover, in comparison to robust apoptosis induced by the approved apoptotic factor FasL, BMP2 only marginally induced cell death. Consistently, neither the known inhibitor of apoptotic cell death zVAD-fmk nor the necroptosis inhibitor necrostatin-1 was able to rescue myeloma cell growth in the presence of BMP2.


Imaging of coronary atherosclerosis - evolution towards new treatment strategies.
Coronary atherosclerosis and the precipitation of acute myocardial infarction are highly complex processes, which makes accurate risk prediction challenging. Rapid developments in invasive and noninvasive imaging technologies now provide us with detailed, exquisite images of the coronary vasculature that allow direct investigation of a wide range of these processes. These modalities include sophisticated assessments of luminal stenoses and myocardial perfusion, complemented by novel measures of the atherosclerotic plaque burden, adverse plaque characteristics, and disease activity. Together, they can provide comprehensive, individualized assessments of coronary atherosclerosis as it occurs in patients. Not only can this information provide important pathological insights, but it can also potentially be used to guide personalized treatment decisions. In this Review, we describe the latest advances in both established and emerging imaging techniques, focusing on the strengths and weakness of each approach. Moreover, we discuss how these technological advances might be translated from attractive images into novel imaging strategies and definite improvements in clinical risk prediction and patient outcomes. This process will not be easy, and the many potential barriers and difficulties are also reviewed.


Visualization of gene expression of short and long forms of prolactin receptor in the rat.
PRL receptor gene expression was visualized in various tissues by in situ hybridization, using 35S-labeled probes unique to each form of receptor. Tissues were removed rapidly from adult male and female rats and placed in liquid nitrogen. Cryostat sections (10 microns) were prepared, fixed, pretreated, and dehydrated before incubation with the various probes. Hybridization was performed overnight, after which the slides were first exposed to autoradiographic film and then dipped in nuclear emulsion and exposed for 1-2 weeks. The specificity of the signal was studied by competition and using radiolabeled heterologous probes. Some tissues show no expression of either form of receptor mRNA, such as olfactory bulb and penis. Macroautoradiogram signals (optical density) were compared to a standard curve to observe the variation in mRNA expression, which was expressed in arbitrary units. Sex differences in the expression of PRL receptors were seen in a number of tissues, such as adrenal gland and pituitary. Expression of mRNAs specific to the long form of PRL receptor was predominant in adrenal gland, pituitary, thymus, spleen, skin, heart, and skeletal muscle, whereas the short form was expressed to a greater extent in kidney and lung. At the light microscopic level, the silver grains observed by epipolarization or light field were seen in the specific regions or cells that express PRL receptor mRNAs. In conclusion, the long form transcript predominates in most tissues, except kidney and lung. The advantage of in situ hybridization is that it allows the identification of specific regions or cells expressing mRNAs to be identified. The actions of PRL have not been identified in all tissues expressing PRL receptor transcripts, nor has any definitive correlation been made with the expression of short and long forms of PRL receptor and function.


Experimental confirmation at visible light wavelengths of the backscattering enhancement phenomenon of the photonic nanojet.
We report what we believe is the first experimental confirmation at visible light wavelengths of the backscattering enhancement phenomenon of the photonic nanojet. A specially designed sample stage consisting of a multilayered sandwich of glass, solid polydimethylsiloxane (PDMS), and liquid PDMS, permitted the precise positioning of a gold nanoparticle of diameter between 50 and 100 nm within the nanojet emitted by a 4.4 μm diameter BaTiO(3) microsphere embedded within the PDMS. We determined that, when the gold nanoparticle is optimally positioned within the nanojet, the backscattering of the microsphere can greatly increase: for example, by 3:1 (200%) for the 50 nm gold nanoparticle. The increased backscattering is strongly dependent upon the illumination wavelength and the numerical aperture of the imaging system, and occurs for nonresonant illuminations of the isolated microsphere. Low objective numerical apertures of approximately 0.075 yield the maximum observed increases in backscattering. The measured data agree well with numerical calculations incorporating Mie-based theory and Fourier optics.


A genetic modifier suggests that endurance exercise exacerbates Huntington's disease.
Polyglutamine expansions in the huntingtin gene cause Huntington's disease (HD). Huntingtin is ubiquitously expressed, leading to pathological alterations also in peripheral organs. Variations in the length of the polyglutamine tract explain up to 70% of the age-at-onset variance, with the rest of the variance attributed to genetic and environmental modifiers. To identify novel disease modifiers, we performed an unbiased mutagenesis screen on an HD mouse model, identifying a mutation in the skeletal muscle voltage-gated sodium channel (Scn4a, termed 'draggen' mutation) as a novel disease enhancer. Double mutant mice (HD; Scn4aDgn/+) had decreased survival, weight loss and muscle atrophy. Expression patterns show that the main tissue affected is skeletal muscle. Intriguingly, muscles from HD; Scn4aDgn/+ mice showed adaptive changes similar to those found in endurance exercise, including AMPK activation, fibre type switching and upregulation of mitochondrial biogenesis. Therefore, we evaluated the effects of endurance training on HD mice. Crucially, this training regime also led to detrimental effects on HD mice. Overall, these results reveal a novel role for skeletal muscle in modulating systemic HD pathogenesis, suggesting that some forms of physical exercise could be deleterious in neurodegeneration.


The effect of carbohydrate moiety structure on the immunoregulatory activity of lactoferrin in vitro.
The aim of this study was to evaluate the immunoregulatory effects of recombinant human lactoferrin (rhLF) in two in vitro models: (1) the secondary humoral immune response to sheep erythrocytes (SRBC); and (2) the mixed lymphocyte reaction (MLR). We compared the non-sialylated glycoform of rhLF as expressed by glycoengineered Pichia pastoris with one that was further chemically sialylated. In an earlier study, we showed that sialylated rhLF could reverse methotrexate-induced suppression of the secondary immune response of mouse splenocytes to SRBC, and that the phenomenon is dependent on the interaction of lactoferrin (LF) with sialoadhesin (CD169). We found that the immunorestorative activity of sialylated rhLF is also dependent on its interaction with the CD22 antigen, a member of the immunoglobulin superfamily that is expressed by B lymphocytes. We also demonstrated that only sialylated rhLF was able to inhibit the MLR reaction. MLR was inhibited by bovine lactoferrin (bLF), a glycoform that has a more complex glycan structure. Desialylated bLF and lactoferricin, a bLF-derived peptide devoid of carbohydrates, did not express such inhibitory activity. We showed that the interaction of LF with sialic acid receptors is essential for at least some of the immunoregulatory activity of this glycoprotein.


Detection of fetal cells in maternal blood.
We report the detection of fetal cells in the maternal circulation by enzymatic amplification of a single copy gene sequence that was fetal-specific. Fetal HLA-A2-positive cells were sorted from maternal HLA-A2-negative cells by flow cytometry and confirmed by demonstration of a fetal-specific HLA-DR4 sequence. However, this sequence could not be detected in unenriched maternal DNA prepared at 28 and 32 weeks' gestation. The sensitivity of detection was 1 HLA-DR4-positive cell in 10(5) HLA-DR4-negative cells. We conclude that prenatal diagnosis of paternally inherited autosomal-dominant genetic defects may be possible by selective gene amplification of maternal peripheral blood. However, preliminary enrichment for fetal cells may be necessary.


Informal urban settlements and cholera risk in Dar es Salaam, Tanzania.
As a result of poor economic opportunities and an increasing shortage of affordable housing, much of the spatial growth in many of the world's fastest-growing cities is a result of the expansion of informal settlements where residents live without security of tenure and with limited access to basic infrastructure. Although inadequate water and sanitation facilities, crowding and other poor living conditions can have a significant impact on the spread of infectious diseases, analyses relating these diseases to ongoing global urbanization, especially at the neighborhood and household level in informal settlements, have been infrequent. To begin to address this deficiency, we analyzed urban environmental data and the burden of cholera in Dar es Salaam, Tanzania. Cholera incidence was examined in relation to the percentage of a ward's residents who were informal, the percentage of a ward's informal residents without an improved water source, the percentage of a ward's informal residents without improved sanitation, distance to the nearest cholera treatment facility, population density, median asset index score in informal areas, and presence or absence of major roads. We found that cholera incidence was most closely associated with informal housing, population density, and the income level of informal residents. Using data available in this study, our model would suggest nearly a one percent increase in cholera incidence for every percentage point increase in informal residents, approximately a two percent increase in cholera incidence for every increase in population density of 1000 people per km(2) in Dar es Salaam in 2006, and close to a fifty percent decrease in cholera incidence in wards where informal residents had minimally improved income levels, as measured by ownership of a radio or CD player on average, in comparison to wards where informal residents did not own any items about which they were asked. In this study, the range of access to improved sanitation and improved water sources was quite narrow at the ward level, limiting our ability to discern relationships between these variables and cholera incidence. Analysis at the individual household level for these variables would be of interest. Our results suggest that ongoing global urbanization coupled with urban poverty will be associated with increased risks for certain infectious diseases, such as cholera, underscoring the need for improved infrastructure and planning as the world's urban population continues to expand.


Feeling bad and looking worse: negative affect is associated with reduced perceptions of face-healthiness.
Some people perceive themselves to look more, or less attractive than they are in reality. We investigated the role of emotions in enhancement and derogation effects; specifically, whether the propensity to experience positive and negative emotions affects how healthy we perceive our own face to look and how we judge ourselves against others. A psychophysical method was used to measure healthiness of self-image and social comparisons of healthiness. Participants who self-reported high positive (N = 20) or negative affectivity (N = 20) judged themselves against healthy (red-tinged) and unhealthy looking (green-tinged) versions of their own and stranger's faces. An adaptive staircase procedure was used to measure perceptual thresholds. Participants high in positive affectivity were un-biased in their face health judgement. Participants high in negative affectivity on the other hand, judged themselves as equivalent to less healthy looking versions of their own face and a stranger's face. Affective traits modulated self-image and social comparisons of healthiness. Face health judgement was also related to physical symptom perception and self-esteem; high physical symptom reports were associated a less healthy self-image and high self-reported (but not implicit) self-esteem was associated with more favourable social comparisons of healthiness. Subject to further validation, our novel face health judgement task could have utility as a perceptual measure of well-being. We are currently investigating whether face health judgement is sensitive to laboratory manipulations of mood.


Identification of circular RNAs with host gene-independent expression in human model systems for cardiac differentiation and disease.
Cardiovascular disease, one of the most common causes of death in western populations, is characterized by changes in RNA splicing and expression. Circular RNAs (circRNA) originate from back-splicing events, which link a downstream 5' splice site to an upstream 3' splice site. Several back-splicing junctions (BSJ) have been described in heart biopsies from human, rat and mouse hearts (Werfel et al., 2016; Jakobi et al., 2016 ). Here, we use human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) to identify circRNA and host gene dynamics in cardiac development and disease. In parallel, we explore candidate interactions of selected homologs in mouse and rat via RIP-seq experiments. Deep RNA sequencing of cardiomyocyte development and β-adrenergic stimulation uncovered 4518 circRNAs. The set of circular RNA host genes is enriched for chromatin modifiers and GTPase activity regulators. RNA-seq and qRT-PCR data showed that circular RNA expression is highly dynamic in the hiPSC-CM model with 320 circRNAs showing significant expression changes. Intriguingly, 82 circRNAs are independently regulated to their host genes. We validated the same circRNA dynamics for circRNAs from ATXN10, CHD7, DNAJC6 and SLC8A1 in biopsy material from human dilated cardiomyopathy (DCM) and control patients. Finally, we could show that rodent homologs of circMYOD, circSLC8A1, circATXN7 and circPHF21A interact with either the ribosome or Argonaute2 protein complexes. CircRNAs are dynamically expressed in a hiPSC-CM model of cardiac development and stress response. Some circRNAs show similar, host-gene independent expression dynamics in patient samples and may interact with the ribosome and RISC complex. In summary, the hiPSC-CM model uncovered a new signature of potentially disease relevant circRNAs which may serve as novel therapeutic targets.


The Prehistoric Indian Ayurvedic Rice Shashtika Is an Extant Early Domesticate With a Distinct Selection History.
Fully domesticated rice is considered to have emerged in India at approximately 2000 B.C., although its origin in India remains a contentious issue. The fast-growing 60-days rice strain described in the Vedic literature (1900-500 B.C.) and termed Shashtika (Sanskrit) or Njavara (Dravidian etymology) in Ayurveda texts including the seminal texts Charaka Samhita and Sushruta Samhita (circa 660-1000 B.C.) is a reliable extant strain among the numerous strains described in the Ayurveda literature. We here report the results of the phylogenetic analysis of Njavara accessions in relation to the cultivars belonging to the known ancestral sub-groups indica, japonica, aromatic, and aus in rice gene pool and the populations of the progenitor species Oryza rufipogon using genetic and gene genealogical methods. Based on neutral microsatellite markers, Njavara produced a major clade, which comprised of minor clades corresponding to the genotypic classes reported in Njavara germplasm, and was distinct from that were produced by the ancestral sub-groups. Further we performed a phylogenetic analysis using the combined sequence of 19 unlinked EST-based sequence tagged site (STS) loci with proven potential in inferring rice phylogeny. In the phylogenetic tree also the Njavara genotypic classes were clearly separated from the ancestral sub-groups. For most loci the genealogical analysis produced a high frequency central haplotype shared among most of the rice samples analyzed in the study including Njavara and a set of O. rufipogon accessions. The haplotypes sharing pattern with the progenitor O. rufipogon suggests a Central India-Southeast Asia origin for Njavara. Results signify that Njavara is genetically distinct in relation to the known ancestral sub-groups in rice. Further, from the phylogenetic features together with the reported morphological characteristics, it is likely that Njavara is an extant early domesticate in Indian rice gene pool, preserved in pure form over millennia by the traditional prudence in on-farm selection using 60-days maturity, because of its medicinal applications.


Temporal integration of movement: the time-course of motion streaks revealed by masking.
Temporal integration in the visual system causes fast-moving objects to leave oriented 'motion streaks' in their wake, which could be used to facilitate motion direction perception. Temporal integration is thought to occur over ≈100 ms in early cortex, although this has never been tested for motion streaks. Here we compare the ability of fast-moving ('streaky') and slow-moving fields of dots to mask briefly flashed gratings either parallel or orthogonal to the motion trajectory. Gratings were presented at various asynchronies relative to motion onset (from -200 to +700 ms) to sample the time-course of the accumulating streaks. Predictions were that masking would be strongest for the fast parallel condition, and would be weak at early asynchronies and strengthen over time as integration rendered the translating dots more streaky and grating-like. The asynchrony where the masking function reached a plateau would correspond to the temporal integration period. As expected, fast-moving dots caused greater masking of parallel gratings than orthogonal gratings, and slow motion produced only modest masking of either grating orientation. Masking strength in the fast, parallel condition increased with time and reached a plateau after 77 ms, providing an estimate of the temporal integration period for mechanisms encoding motion streaks. Interestingly, the greater masking by fast motion of parallel compared with orthogonal gratings first reached significance at 48 ms before motion onset, indicating an effect of backward masking by motion streaks.


Cocaine Use in the Infertile Male Population: A Marker for Conditions Resulting in Subfertility.
We sought to evaluate the incidence and effect of cocaine use in the infertile male population. Men presenting for fertility evaluation reporting cocaine usage were identified via prospectively collected database. Data were analyzed for usage patterns, reproductive history, associated drug use and medical conditions, hormonal and semen parameters. Thirty-eight out of 4,400 (0.9%) men reported cocaine use. Most used cocaine every 3 months or less. Compared with non-cocaine using men, cocaine users reported more recreational drug use (89 vs. 9.2%), marijuana use (78.9 vs. 11.4%), chlamydia (10.5 vs. 3%), herpes (7.9 vs. 2.5%), and tobacco use (55.3 vs. 19.5%). After excluding men with causes for azoospermia, the mean semen parameters for cocaine users were: volume 2.47 ± 1.02 ml; concentration 53.55 ± 84.04 × 10(6)/ml; motility 15.72 ± 12.26%; total motile sperm count 76.67 ± 180.30 × 10(6). Few (< 1%) men in our infertile population reported the use of cocaine, and the frequency of use was low. Given the low use rates and limitations of reporting bias, it is difficult to determine the direct effect of cocaine use on male fertility. However, while infrequent cocaine use seems to have limited impact on semen parameters, men reporting cocaine use represent a different cohort of men than the overall infertile population, with higher rates of concurrent substance abuse, tobacco use and infections, all of which may negatively impact their fertility. Reported cocaine users should be screened for concurrent drug use and infections.


Synergistic cytotoxic effects of bortezomib and CK2 inhibitor CX-4945 in acute lymphoblastic leukemia: turning off the prosurvival ER chaperone BIP/Grp78 and turning on the pro-apoptotic NF-κB.
The proteasome inhibitor bortezomib is a new targeted treatment option for refractory or relapsed acute lymphoblastic leukemia (ALL) patients. However, a limited efficacy of bortezomib alone has been reported. A terminal pro-apoptotic endoplasmic reticulum (ER) stress/unfolded protein response (UPR) is one of the several mechanisms of bortezomib-induced apoptosis. Recently, it has been documented that UPR disruption could be considered a selective anti-leukemia therapy. CX-4945, a potent casein kinase (CK) 2 inhibitor, has been found to induce apoptotic cell death in T-ALL preclinical models, via perturbation of ER/UPR pathway. In this study, we analyzed in T- and B-ALL preclinical settings, the molecular mechanisms of synergistic apoptotic effects observed after bortezomib/CX-4945 combined treatment. We demonstrated that, adding CX-4945 after bortezomib treatment, prevented leukemic cells from engaging a functional UPR in order to buffer the bortezomib-mediated proteotoxic stress in ER lumen. We documented that the combined treatment decreased pro-survival ER chaperon BIP/Grp78 expression, via reduction of chaperoning activity of Hsp90. Bortezomib/CX-4945 treatment inhibited NF-κB signaling in T-ALL cell lines and primary cells from T-ALL patients, but, intriguingly, in B-ALL cells the drug combination activated NF-κB p65 pro-apoptotic functions. In fact in B-cells, the combined treatment induced p65-HDAC1 association with consequent repression of the anti-apoptotic target genes, Bcl-xL and XIAP. Exposure to NEMO (IKKγ)-binding domain inhibitor peptide reduced the cytotoxic effects of bortezomib/CX-4945 treatment. Overall, our findings demonstrated that CK2 inhibition could be useful in combination with bortezomib as a novel therapeutic strategy in both T- and B-ALL.


Assessing the acoustic behaviour of Anopheles gambiae (s.l.) dsxF mutants: implications for vector control.
Release of gene-drive mutants to suppress Anopheles mosquito reproduction is a promising method of malaria control. However, many scientific, regulatory and ethical questions remain before transgenic mosquitoes can be utilised in the field. At a behavioural level, gene-drive carrying mutants should be at least as sexually attractive as the wildtype populations they compete against, with a key element of Anopheles copulation being acoustic courtship. We analysed sound emissions and acoustic preference in a doublesex mutant previously used to collapse Anopheles gambiae (s.l.) cages. Anopheles rely on flight tones produced by the beating of their wings for acoustic mating communication. We assessed the impact of disrupting a female-specific isoform of the doublesex gene (dsxF) on the wing beat frequency (WBF; measured as flight tone) of males (XY) and females (XX) in homozygous dsxF- mutants (dsxF-/-), heterozygous dsxF- carriers (dsxF+/-) and G3 dsxF+ controls (dsxF+/+). To exclude non-genetic influences, we controlled for temperature and wing length. We used a phonotaxis assay to test the acoustic preferences of mutant and control mosquitoes. A previous study showed an altered phenotype only for dsxF-/- females, who appear intersex, suggesting that the female-specific dsxF allele is haplosufficient. We identified significant, dose-dependent increases in the WBF of both dsxF-/- and dsxF+/- females compared to dsxF+/+ females. All female WBFs remained significantly lower than male equivalents, though. Males showed stronger phonotactic responses to the WBFs of control dsxF+/+ females than to those of dsxF+/- and dsxF-/- females. We found no evidence of phonotaxis in any female genotype. No male genotypes displayed any deviations from controls. A prerequisite for anopheline copulation is the phonotactic attraction of males towards female flight tones within mating swarms. Reductions in mutant acoustic attractiveness diminish their mating efficiency and thus the efficacy of population control efforts. Caged population assessments may not successfully reproduce natural mating scenarios. We propose to amend existing testing protocols to better reflect competition between mutants and target populations. Our findings confirm that dsxF disruption has no effect on males; for some phenotypic traits, such as female WBFs, the effects of dsxF appear dose-dependent rather than haplosufficient.


Vitamin E and C supplementation and risk of cancer in men: posttrial follow-up in the Physicians' Health Study II randomized trial.
Recent posttrial analysis of a completed randomized trial found an increased risk of prostate cancer among healthy men taking high-dose vitamin E supplements. Trials that examined the effect of vitamin C supplements on cancer risk are few. We examined whether vitamin E or vitamin C supplementation affects the risk of cancer events during posttrial follow-up of the Physicians' Health Study II. Beginning in 1997, a total of 14,641 US male physicians aged ≥50 y were randomly assigned to receive 400 IU of vitamin E every other day, 500 mg of vitamin C daily, or their respective placebos. The vitamin E and vitamin C treatment ended in 2007, and observational follow-up continued through June 2011. This study included an additional 356 cases of incident prostate cancer and 771 total cancers that developed during a mean (maximum) of 2.8 (3.8) y of posttrial observation. During an overall mean of 10.3 (13.8) y, there were a total of 1373 incident prostate cancers and 2669 total cancers documented. In comparison with placebo, vitamin E supplementation had no effect on the incidence of prostate cancer (HR: 0.99; 95% CI: 0.89, 1.10) or total cancers (HR: 1.02; 95% CI: 0.95, 1.10). There was also no effect of vitamin C supplementation on total cancers (HR: 1.02; 95% CI: 0.94, 1.10) or incident prostate cancer (HR: 1.03; 95% CI: 0.93, 1.15). Neither vitamin E nor vitamin C supplementation had effects on other site-specific cancers overall. Stratification by known cancer risk factors, history of cancer, other randomized treatment, and follow-up time showed no significant interactions. In this large-scale randomized trial in men, vitamin E and C supplementation had no immediate or long-term effects on the risk of total cancers, prostate cancer, or other site-specific cancers.


Multiple charged and aromatic residues in CCR5 amino-terminal domain are involved in high affinity binding of both chemokines and HIV-1 Env protein.
CCR5 is a functional receptor for MIP-1alpha, MIP-1beta, RANTES (regulated on activation normal T cell expressed), MCP-2, and MCP-4 and constitutes the main coreceptor for macrophage tropic human and simian immunodeficiency viruses. By using CCR5-CCR2b chimeras, we have shown previously that the second extracellular loop of CCR5 is the major determinant for chemokine binding specificity, whereas the amino-terminal domain plays a major role for human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus coreceptor function. In the present work, by using a panel of truncation and alanine-scanning mutants, we investigated the role of specific residues in the CCR5 amino-terminal domain for chemokine binding, functional response to chemokines, HIV-1 gp120 binding, and coreceptor function. Truncation of the amino-terminal domain resulted in a progressive decrease of the binding affinity for chemokines, which correlated with a similar drop in functional responsiveness. Mutants lacking residues 2-13 exhibited fairly weak responses to high concentrations (500 nM) of RANTES or MIP-1beta. Truncated mutants also exhibited a reduction in the binding affinity for R5 Env proteins and coreceptor activity. Deletion of 4 or 12 residues resulted in a 50 or 80% decrease in coreceptor function, respectively. Alanine-scanning mutagenesis identified several charged and aromatic residues (Asp-2, Tyr-3, Tyr-10, Asp-11, and Glu-18) that played an important role in both chemokine and Env high affinity binding. The overlapping binding site of chemokines and gp120 on the CCR5 amino terminus, as well as the involvement of these residues in the epitopes of monoclonal antibodies, suggests that these regions are particularly exposed at the receptor surface.


Contrasting patterns of gene expression indicate differing pyrethroid resistance mechanisms across the range of the New World malaria vector Anopheles albimanus.
Decades of unmanaged insecticide use and routine exposure to agrochemicals have left many populations of malaria vectors in the Americas resistant to multiple classes of insecticides, including pyrethroids. The molecular basis of pyrethroid resistance is relatively uncharacterised in American malaria vectors, preventing the design of suitable resistance management strategies. Using whole transcriptome sequencing, we characterized the mechanisms of pyrethroid resistance in Anopheles albimanus from Peru and Guatemala. An. albimanus were phenotyped as either deltamethrin or alpha-cypermethrin resistant. RNA from 1) resistant, 2) unexposed, and 3) a susceptible laboratory strain of An. albimanus was sequenced and analyzed using RNA-Seq. Expression profiles of the three groups were compared based on the current annotation of the An. albimanus reference genome. Several candidate genes associated with pyrethroid resistance in other malaria vectors were found to be overexpressed in resistant An. albimanus. In addition, gene ontology terms related to serine-type endopeptidase activity, extracellular activity and chitin metabolic process were also commonly overexpressed in the field caught resistant and unexposed samples from both Peru and Guatemala when compared to the susceptible strain. The cytochrome P450 CYP9K1 was overexpressed 14x in deltamethrin and 8x in alpha-cypermethrin-resistant samples from Peru and 2x in deltamethrin-resistant samples from Guatemala, relative to the susceptible laboratory strain. CYP6P5 was overexpressed 68x in deltamethrin-resistant samples from Peru but not in deltamethrin-resistant samples from Guatemala. When comparing overexpressed genes between deltamethrin-resistant and alpha-cypermethrin-resistant samples from Peru, a single P450 gene, CYP4C26, was overexpressed 9.8x (p<0.05) in alpha-cypermethrin-resistant samples. In Peruvian deltamethrin-resistant samples, the knockdown resistance mutation (kdr) variant alleles at position 1014 were rare, with approximately 5% frequency, but in the alpha-cypermethrin-resistant samples, the frequency of these alleles was approximately 15-30%. Functional validation of the candidate genes and the kdr mutation as a resistance marker for alpha-cypermethrin will confirm the role of these mechanisms in conferring pyrethroid resistance.


A rare case of typhoid presenting with fever, ascites, hyponatremia, thrombocytopenia, mesenteric lymphadenitis, and multi-drug resistance.
A rare case of typhoid presenting with thrombocytopenia, hyponatremia, ascites mesenteric adenitis, and multi-drug resistance is being presented in this article. An 8-year-old girl was admitted with a history of fever, vomiting, abdominal pain and loose stools. Clinical examination revealed fever and hepatosplenomegaly. Investigations showed leucopenia, thrombocytopenia and hyponatremia. Blood Widal was positive, and blood culture grew Salmonella typhi. Ultrasound abdomen revealed ascites, hepatosplenomegaly, mesenteric lymphadenopathy and thickening of the gall bladder. She was treated with ciprofloxacin intravenously for 6 days and when the fever persisted injection ceftriaxone was added. Ciprofloxacin was given intravenously for a total of 15 days and injection ceftriaxone was given for 12 days. Even then, the fever persisted and hence oral azithromycin was added. Fever subsided completely in 3 days with azithromycin and she became asymptomatic without fever, loose stools, abdominal pain or anything on follow-up after 3 months.


Assessment of Antimicrobial Effectiveness of Neem and Clove Extract Against Streptococcus mutans and Candida albicans: An In vitro Study.
There is increasing interest to develop antimicrobial aids from alternative sources such as medicinal plants for the treatment of infectious diseases. Neem and clove are known to have antimicrobial properties. The study aimed at detecting the antibacterial and antifungal activity of neem and clove extract against Streptococcus mutans and Candida albicans. Strains of S. mutans and C. albicans and selective media for growing micro-organisms were procured. Antimicrobial activity was assessed using two methods, by determining the minimum inhibitory concentration (MIC) using the broth dilution method and determining the zone of inhibition using well diffusion method on mitis salivarius bacitracin selective for S. mutans and Saboraud's dextrose agar plates for C. albicans. One way ANOVA with post hoc analysis was done to compare the antimicrobial activity of extracts and 0.2% chlorhexidine. MIC of neem extract was found to be 4.2 mg/ml and 5.0 mg/ml against S. mutans and C. albicans, respectively. While for cloves, it was 5.5 mg/ml for both. Neem had the highest antibacterial activity with a mean zone of inhibition of 11.4 mm followed by chlorhexidine and cloves whereas antifungal activity was highest for chlorhexidine (14.4 mm) followed by neem and clove. The result of the study established that both plant extracts possess antimicrobial activity against common microbes present in the oral cavity.