APOL1 Risk Variants, Acute Kidney Injury, and Death in Participants With African Ancestry Hospitalized With COVID-19 From the Million Veteran Program.
Coronavirus disease 2019 (COVID-19) confers significant risk of acute kidney injury (AKI). Patients with COVID-19 with AKI have high mortality rates. Individuals with African ancestry with 2 copies of apolipoprotein L1 (APOL1) variants G1 or G2 (high-risk group) have significantly increased rates of kidney disease. We tested the hypothesis that the APOL1 high-risk group is associated with a higher-risk of COVID-19-associated AKI and death. This retrospective cohort study included 990 participants with African ancestry enrolled in the Million Veteran Program who were hospitalized with COVID-19 between March 2020 and January 2021 with available genetic information. The primary exposure was having 2 APOL1 risk variants (RV) (APOL1 high-risk group), compared with having 1 or 0 risk variants (APOL1 low-risk group). The primary outcome was AKI. The secondary outcomes were stages of AKI severity and death. Multivariable logistic regression analyses adjusted for preexisting comorbidities, medications, and inpatient AKI risk factors; 10 principal components of ancestry were performed to study these associations. We performed a subgroup analysis in individuals with normal kidney function prior to hospitalization (estimated glomerular filtration rate ≥60 mL/min/1.73 m2). Of the 990 participants with African ancestry, 905 (91.4%) were male with a median (IQR) age of 68 (60-73) years. Overall, 392 (39.6%) patients developed AKI, 141 (14%) developed stages 2 or 3 AKI, 28 (3%) required dialysis, and 122 (12.3%) died. One hundred twenty-five (12.6%) of the participants were in the APOL1 high-risk group. Patients categorized as APOL1 high-risk group had significantly higher odds of AKI (adjusted odds ratio [OR], 1.95; 95% CI, 1.27-3.02; P = .002), higher AKI severity stages (OR, 2.03; 95% CI, 1.37-2.99; P < .001), and death (OR, 2.15; 95% CI, 1.22-3.72; P = .007). The association with AKI persisted in the subgroup with normal kidney function (OR, 1.93; 95% CI, 1.15-3.26; P = .01). Data analysis was conducted between February 2021 and April 2021. In this cohort study of veterans with African ancestry hospitalized with COVID-19 infection, APOL1 kidney risk variants were associated with higher odds of AKI, AKI severity, and death, even among individuals with prior normal kidney function.


RNA G-Quadruplexes in the model plant species Arabidopsis thaliana: prevalence and possible functional roles.
Tandem stretches of guanines can associate in hydrogen-bonded arrays to form G-quadruplexes, which are stabilized by K(+) ions. Using computational methods, we searched for G-Quadruplex Sequence (GQS) patterns in the model plant species Arabidopsis thaliana. We found ∼ 1200 GQS with a G(3) repeat sequence motif, most of which are located in the intergenic region. Using a Markov modeled genome, we determined that GQS are significantly underrepresented in the genome. Additionally, we found ∼ 43,000 GQS with a G(2) repeat sequence motif; notably, 80% of these were located in genic regions, suggesting that these sequences may fold at the RNA level. Gene Ontology functional analysis revealed that GQS are overrepresented in genes encoding proteins of certain functional categories, including enzyme activity. Conversely, GQS are underrepresented in other categories of genes, notably those for non-coding RNAs such as tRNAs and rRNAs. We also find that genes that are differentially regulated by drought are significantly more likely to contain a GQS. CD-detected K(+) titrations performed on representative RNAs verified formation of quadruplexes at physiological K(+) concentrations. Overall, this study indicates that GQS are present at unique locations in Arabidopsis and that folding of RNA GQS may play important roles in regulating gene expression.


Identification of poultry meat-derived fatty acids functioning as quorum sensing signal inhibitors to autoinducer-2 (AI-2).
Autoinducer-2 (AI-2) is a compound that plays a key role in bacterial cell-to-cell communication (quorum sensing). Previous research has shown certain food matrices inhibit this signaling compound. Using the reporter strain, Vibrio harveyi BB170, quorum-sensing inhibitors contained in poultry meat wash (PMW) samples were characterized by molecular weight and hydrophobic properties using liquid chromatography systems. Most fractions that demonstrated AI-2 inhibition were 13.7 kDa or less, and had hydrophobic properties. Hexane was used to extract inhibitory compounds from a PMW preparation and the extract was further separated by gas chromatography (GC). Several fatty acids were identified and quantified. Linoleic acid, oleic acid, palmitic acid, and stearic acid were each tested for inhibition at 0.1, 1, and 10 mM concentrations. All samples expressed AI-2 inhibition (ranging from approximately 25% to 99%). Fatty acids, combined in concentrations equivalent to those determined by GC analysis, expressed inhibition at 59.5%, but higher combined concentrations (10- and 100-fold) had inhibition at 84.4% and 69.5%, respectively. The combined fatty acids (100-fold) did not demonstrate a substantial decrease in colony plate counts, despite presenting high AI-2 inhibition. These fatty acids, through modulating quorum sensing by inhibition, may offer a unique means to control foodborne pathogens and reduce microbial spoilage.


Reproducibility of tricuspid regurgitant jet velocity measurements in children and young adults with sickle cell disease undergoing screening for pulmonary hypertension.
The reproducibility of tricuspid regurgitant jet velocity (TRJV) measurements by Doppler echocardiography has not been subjected to systematic evaluation among individuals with sickle cell disease (SCD) undergoing screening for pulmonary hypertension. We examined sources of disagreement associated with peak TRJV in children and young adults with SCD. Peak TRJV was independently measured and interpreted a week apart by separate sonographers and readers, respectively, in 30 subjects (mean age, 15.8 ± 3.3 years) who provided 120 observations. We assessed intra-/inter-reader, intra-/inter-sonographer, sonographer-reader, and within subject agreement using Intraclass Correlation Coefficient (ICC) and Cohen's kappa (κ). Agreement was examined graphically using Bland-Altman plots. Although sonographers could estimate and measure peak TRJV in all subjects, readers designated tricuspid regurgitation nonquantifiable in 10-17% of their final interpretations. Intra-reader agreement was highest (ICC = 0.93 [95% CI 0.86, 0.97], P = 0.0001) and within subject agreement lowest (ICC = 0.36 [95% CI 0.02, 0.64], P = 0.021) for single TRJV measurements. Similarly, intra-reader agreement was highest (κ = 0.74 [95% CI 0.53, 0.95], P = 0.0001) and within subject lowest (κ = 0.14 [95% CI -0.17, 0.46], P = 0.38) when sonographers and readers categorized TRJV measurements. On Bland-Altman plots, absolute differences in observations increased with higher mean TRJV readings for intra-/inter-reader agreement. Peak TRJV measurements in individual children and young adults with SCD are affected by several sources of disagreement, underscoring the need for methodological improvements that ensure reproducibility of this screening modality for making clinical decisions in this population.


Association between current medication use and progression of radiographic knee osteoarthritis: data from the osteoarthritis initiative.
Use of specific medications may accelerate the progression of radiographic knee OA (RKOA). Our aim was to examine the effect of medication use on the progression of RKOA. We used longitudinal data from the Osteoarthritis Initiative (OAI), an observational study of risk factors for knee OA. At baseline, we selected participants with RKOA (Kellgren-Lawrence grade ≥2) and excluded those with a history of knee-related injury/surgery and other musculoskeletal disorders. Current medication use (use/non-use in the previous 30 days) and radiographic medial minimum joint space width (mJSW) data were available at baseline and annually up to 96 months follow-up. We used random effects, panel regression to assess the association between current medication use (non-users as reference group) and change in mJSW. Of 2054 eligible participants, 2003 participants with baseline mJSW data were included [55.7% female, mean age 63.3 (s.d. 8.98) years]. Of seven medication classes, at baseline NSAIDs were the most frequently used analgesia (14.7%), anti-histamine (10.4%) use was frequent and the following comorbidity medications were used most frequently: statins (27.4%), anti-hypertensives (up to 15.0%), anti-depressant/anxiolytics/psychotropics (14.0%), osteoporosis-related medication (10.9%) and diabetes-related medication (6.9%). Compared with current non-users, current use of NSAIDs was associated with a loss of mJSW (b = -0.042, 95% CI -0.08, -0.0004). No other associations were observed. In current users of NSAIDs, mJSW loss was increased compared with current non-users in participants with RKOA. Clinical trials are required to assess the potential disease-modifying effects of these medications.


Clinical and economic considerations in the treatment of acute exacerbations of chronic bronchitis.
Limited data exist to guide physicians in the cost-effective treatment of acute exacerbation of chronic bronchitis (AECB). Therefore, the main objective of this study was to determine the antimicrobial efficacy and related costs for patients with AECB. A retrospective review of 60 outpatient medical records with a diagnosis of chronic obstructive pulmonary disease (COPD) and chronic bronchitis episodes from a pulmonary clinic of a teaching institution was undertaken. The participating patients had a total of 224 episodes of AECB requiring antibiotic treatment. Before review, empirical antibiotic choices were divided into first-line (amoxycillin, co-trimoxazole, tetracyclines, erythromycin), second-line (cephradine, cefuroxime, cefaclor, cefprozil) and third-line (co-amoxiclav, azithromycin, ciprofloxacin) agents. Patients receiving first-line agents failed significantly more frequently than third-line agents (19% vs 7%, P < 0.05). Additionally, patients prescribed first-line agents were hospitalized significantly more often for AECB within 2 weeks of outpatient treatment as compared with patients prescribed third-line agents (18.0% vs 5.3% third-line agents; P < 0.02). Time between subsequent AECB episodes requiring treatment was significantly longer for patients receiving third-line agents compared with first-line and second-line agents (P < 0.005). Pharmacy costs were lowest with first-line agents (first-line US$10.30 +/- 8.76; second-line US$24.45 +/- 25.65; third-line US$45.40 +/- 11.11; P < 0.0001), but third-line agents showed a trend towards lower mean total costs of AECB treatment (first-line US$942 +/- 2173; second-line, US$563 +/- 2296; third-line, US$542 +/- 1946). The use of third-line antimicrobials, co-amoxiclav, ciprofloxacin or azithromycin, significantly reduced the failure rate and need for hospitalization, prolonged the time between AECB episodes, and showed a lower total cost for the management of AECB. Prospective studies are needed to confirm these findings.


The effect of acrylamide and other sulfhydryl alkylators on the ability of dynein and kinesin to translocate microtubules in vitro.
Chronic exposure to acrylamide leads to a dying-back axonopathy afflicting the longest axons of all tested mammalian and avian species. Prior to the onset of acrylamide-induced axonal degeneration, alterations in axonal fast transport have been consistently reported to be more severe for the retrograde than the anterograde direction. The putative retrograde motor protein, dynein, is compromised by exposure to the sulfhydryl-alkylating agent N-ethylmaleimide (NEM) at concentrations far below those required to inactivate kinesin, the putative anterograde motor protein. Since acrylamide is capable of alkylating protein sulfhydryl moieties, we tested whether a direct exposure of purified kinesin or dynein to acrylamide would result in an impairment of either enzyme's ability to translocate microtubules. Motor activity was assayed by sequentially adsorbing either kinesin or dynein to acid-washed coverslips, treating with an alkylating agent or control solution, adding microtubules and ATP, and finally imaging and quantifying the binding and gliding of microtubules using video-enhanced differential interference contrast (VE-DIC) microscopy. In comparison to controls, incubation of dynein with NEM, ethacrynic acid, or iodoacetic acid resulted in dose-dependent decreases in the amount and rate of microtubule gliding, but increases in irreversible high-affinity microtubule binding. In contrast, exposure of dynein to 1-100 mM solutions of acrylamide did not significantly alter either the binding or gliding of microtubules (a molar/hour exposure to acrylamide equivalent to 50 times that which causes retrograde transport deficits in vivo). Likewise, kinesin motility parameters were not significantly affected by acrylamide concentrations up to 100 mM while NEM solutions > 100 microM led to significant losses in the ability of kinesin to bind MT. These data indicate that acrylamide does not significantly interact with bound (adsorbed) kinesin or dynein, implying that the mechanism by which acrylamide interferes with fast axonal transport in vivo is by interaction with other factor(s) that govern the movement of vesicles.


Gene Regulatory Interactions at Lamina-Associated Domains.
The nuclear lamina provides a repressive chromatin environment at the nuclear periphery. However, whereas most genes in lamina-associated domains (LADs) are inactive, over ten percent reside in local euchromatic contexts and are expressed. How these genes are regulated and whether they are able to interact with regulatory elements remain unclear. Here, we integrate publicly available enhancer-capture Hi-C data with our own chromatin state and transcriptomic datasets to show that inferred enhancers of active genes in LADs are able to form connections with other enhancers within LADs and outside LADs. Fluorescence in situ hybridization analyses show proximity changes between differentially expressed genes in LADs and distant enhancers upon the induction of adipogenic differentiation. We also provide evidence of involvement of lamin A/C, but not lamin B1, in repressing genes at the border of an in-LAD active region within a topological domain. Our data favor a model where the spatial topology of chromatin at the nuclear lamina is compatible with gene expression in this dynamic nuclear compartment.


Comparative microbial source tracking methods for identification of fecal contamination sources at Sunnyside Beach in the Toronto region area of concern.
Several beaches within the Toronto region area of concern have persistent issues with fecal contamination, causing a beach beneficial use impairment (BUI). In this study, Escherichia coli, including ampicillin-resistant strains, were enumerated via culturable and quantitative polymerase chain reaction (qPCR) methods. Microbial source tracking (MST) markers (for general Bacteroidales, human, ruminant/cow, gull, and dog) were detected and enumerated via PCR and qPCR to identify sources of fecal contamination at Sunnyside Beach and in the Humber River. Human, cow, and dog markers had good host-specificity, while gull markers sometimes amplified a few other bird species. The ruminant endpoint PCR marker amplified a variety of other animal species rendering it less useful. Both human and gull fecal contamination were prevalent in the Humber River, while Sunnyside Beach was predominantly impacted by gull fecal contamination. Human sewage impacts were more prevalent in the lower Humber River, particularly in Black Creek. However, to reduce Sunnyside beach postings, reducing bird fecal contamination in the river and at the beach would be necessary. When there are high levels of E. coli throughout a beachshed, an MST toolbox approach can add value to discriminate source(s) of E. coli contamination and guide decisions relating to public health risk and remediation strategies.


Testosterone levels influence mouse fetal Leydig cell progenitors through notch signaling.
Leydig cells are the steroidogenic lineage of the mammalian testis that produces testosterone, a key hormone required throughout male fetal and adult life for virilization and spermatogenesis. Both fetal and adult Leydig cells arise from a progenitor population in the testis interstitium but are thought to be lineage-independent of one another. Genetic evidence indicates that Notch signaling is required during fetal life to maintain a balance between differentiated Leydig cells and their progenitors, but the elusive progenitor cell type and ligands involved have not been identified. In this study, we show that the Notch pathway signals through the ligand JAG1 in perivascular interstitial cells during fetal life. In the early postnatal testis, we show that circulating levels of testosterone directly affect Notch signaling, implicating a feedback role for systemic circulating factors in the regulation of progenitor cells. Between Postnatal Days 3 and 21, as fetal Leydig cells disappear from the testis and are replaced by adult Leydig cells, the perivascular population of interstitial cells active for Notch signaling declines, consistent with distinct regulation of adult Leydig progenitors.


Information content of colored motifs in complex networks.
We study complex networks in which the nodes are tagged with different colors depending on their function (colored graphs), using information theory applied to the distribution of motifs in such networks. We find that colored motifs can be viewed as the building blocks of the networks (much more than the uncolored structural motifs can be) and that the relative frequency with which these motifs appear in the network can be used to define its information content. This information is defined in such a way that a network with random coloration (but keeping the relative number of nodes with different colors the same) has zero color information content. Thus, colored motif information captures the exceptionality of coloring in the motifs that is maintained via selection. We study the motif information content of the C. elegans brain as well as the evolution of colored motif information in networks that reflect the interaction between instructions in genomes of digital life organisms. While we find that colored motif information appears to capture essential functionality in the C. elegans brain (where the color assignment of nodes is straightforward), it is not obvious whether the colored motif information content always increases during evolution, as would be expected from a measure that captures network complexity. For a single choice of color assignment of instructions in the digital life form Avida, we find rather that colored motif information content increases or decreases during evolution, depending on how the genomes are organized, and therefore could be an interesting tool to dissect genomic rearrangements.


Helicobacter pylori heat-shock protein 60 induces inflammatory responses through the Toll-like receptor-triggered pathway in cultured human gastric epithelial cells.
Contact between Helicobacter pylori and gastric epithelial cells results in activation of NF-kappaB followed by secretion of interleukin (IL)-8. However, host-cell receptor(s) and their ligands involved in H. pylori-related IL-8 production have yet to be fully defined. In this study, the interaction between Toll-like receptors (TLRs), which are host receptors for pathogens involved in the innate immune response, and heat-shock protein (HSP) 60, an immune-potent antigen of H. pylori, was examined during H. pylori-induced IL-8 secretion in vitro. Recombinant H. pylori HSP60 (rHpHSP60) was prepared and added to cultured KATO III human gastric epithelial cells with or without pre-incubation with mouse monoclonal anti-TLR2 or anti-TLR4 antibodies. IL-8 mRNA expression and IL-8 protein release were analysed by Northern blotting and immunoassay. Involvement of NF-kappaB activation was analysed immunocytochemically by anti-NF-kappaB p65 antibody and ammonium pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-kappaB-mediated transcriptional activation. rHpHSP60 induced IL-8 mRNA expression and IL-8 secretion in a dose-dependent manner in KATO III cells. Anti-TLR2 antibody inhibited rHpHSP60-induced IL-8 secretion by 75 %, and anti-TLR4 antibody inhibited it by 30 %. rHpHSP60 induced nuclear translocation of NF-kappaB p65, which was inhibited by pretreatment with anti-TLR2 antibody. Treatment with PDTC significantly decreased the secretion of IL-8 induced by rHpHSP60. These findings suggest that H. pylori HSP60 activates NF-kappaB and induces IL-8 production through TLR-triggered pathways in gastric epithelial cells. Thus, it is possible that H. pylori HSP60 and TLR interaction in host cells contributes to the development of gastric inflammation caused by H. pylori infection.


Pseudomonas aeruginosa induced lung injury model.
In order to study human acute lung injury and pneumonia, it is important to develop animal models to mimic various pathological features of this disease. Here we have developed a mouse lung injury model by intra-tracheal injection of bacteria Pseudomonas aeruginosa (P. aeruginosa or PA). Using this model, we were able to show lung inflammation at the early phase of injury. In addition, alveolar epithelial barrier leakiness was observed by analyzing bronchoalveolar lavage (BAL); and alveolar cell death was observed by Tunel assay using tissue prepared from injured lungs. At a later phase following injury, we observed cell proliferation required for the repair process. The injury was resolved 7 days from the initiation of P. aeruginosa injection. This model mimics the sequential course of lung inflammation, injury and repair during pneumonia. This clinically relevant animal model is suitable for studying pathology, mechanism of repair, following acute lung injury, and also can be used to test potential therapeutic agents for this disease.


Classifications in psychiatry: a conceptual history.
Historical accounts of psychiatric classifications have hitherto been written in terms of a 'received view'. This contains two assumptions, that: (i) the activity of classifying is inherent to the human mind; and (ii) psychiatric 'phenomena' are stable natural objects. The aim of this article is to provide an outline of the evolution of psychiatric classifications from the perspective of conceptual history. This is defined as a theoretical and empirical inquiry into the principles, sortal techniques and contexts in which alienists carried out their task. It assumes that all psychiatric classifications are cultural products, and endeavours to answer the question of whether classificatory models imported from the natural sciences can be applied to man-made constructs (such as mental illness) definitionally based on 'personalised semantics'. Exemplars of classificatory activity are first mapped and contextualised. Then, it is suggested that in each historical period crafting classifications has been like playing a game of chess with each move being governed by rules. This is illustrated by offering an analysis of the 1860-1861 French debate on classification. (1) Medicine is not a contemplative but a modificatory activity and hence classifications are only valuable if they can release new information about the object classified. (2) It should not be inferred from the fact that psychiatric classifications are not working well (i.e. that they only behave as actuarial devices) that they must be given up. Conceptual work needs to continue to identify 'invariants' (i.e. stable elements that anchor classifications to 'nature'. (3) Because mental disorders are more than unstable behavioural epiphenomena wrapped around stable molecular changes, 'neurobiological' invariants may not do. Stability depends upon time frames. Furthermore, it is unlikely that gene-based classifications will ever be considered as classifications of mental disorders. For once, they would have low predictive power because of their lack of information about the defining codes of mental illness. 'Social' and 'psychological' invariants have problems of their own.


Bayesian nonparametric hierarchical modeling.
In biomedical research, hierarchical models are very widely used to accommodate dependence in multivariate and longitudinal data and for borrowing of information across data from different sources. A primary concern in hierarchical modeling is sensitivity to parametric assumptions, such as linearity and normality of the random effects. Parametric assumptions on latent variable distributions can be challenging to check and are typically unwarranted, given available prior knowledge. This article reviews some recent developments in Bayesian nonparametric methods motivated by complex, multivariate and functional data collected in biomedical studies. The author provides a brief review of flexible parametric approaches relying on finite mixtures and latent class modeling. Dirichlet process mixture models are motivated by the need to generalize these approaches to avoid assuming a fixed finite number of classes. Focusing on an epidemiology application, the author illustrates the practical utility and potential of nonparametric Bayes methods.


The Parent-Child Sleep Interactions Scale (PSIS) for preschoolers: factor structure and initial psychometric properties.
Given the high prevalence of sleep problems in early childhood and the significant impact of parenting on children's sleep, the current study aimed to determine the factor structure and psychometric properties of a new measure for assessing sleep-related parenting behaviors and interactions among preschool-aged children-The Parent-Child Sleep Interactions Scale (PSIS). Data was collected among parents of 209 preschool-aged children in two diverse metropolitan areas. All parents completed demographic questionnaires, the Parent-Child Sleep Interaction Scale (PSIS), and the Child Behavior Checklist (CBCL). A subset of parents completed structured interviews using the Preschool Age Psychiatric Assessment (PAPA). Following data reduction procedures, exploratory factor analysis (EFA) using principal axis extraction and oblique rotation was conducted, and internal consistency was assessed. Associations between PSIS scores and sleep problems based on the CBCL and PAPA as well as child sleep problems during infancy were examined. Differences based on demographic variables including race/ethnicity were also investigated. EFA revealed a three-factor solution explaining 60% of the variance in total PSIS scores. Individual factors based on 12 items were labeled Sleep Reinforcement, Sleep Conflict, and Sleep Dependence. Internal consistency for all subscales and total PSIS scores was acceptable. PSIS subscales were positively correlated with both CBCL and PAPA Sleep Problems. Sleep problems during the first year of life were associated with Sleep Conflict and total PSIS scores. Significant differences in PSIS scores based on race/ ethnicity were found. The PSIS shows promise as a valid measure of sleep-related parent/child behaviors and interactions among preschoolers.


Engaging School and Family in Navajo Gardening for Health: Development of the Yéego Intervention to Promote Healthy Eating among Navajo Children.
Navajo children are at increased risk for obesity, in part due to limited access to healthy foods. School garden interventions have been shown to increase access to fresh fruit and vegetables and consumption of healthy foods. Our study describes the development and pilot testing of a school garden intervention for Navajo elementary school children. We reviewed existing school garden interventions and conducted formative research with students, caregivers, and school staff to inform the intervention. The intervention consisted of a garden built at the school and a yearlong curriculum on gardening and healthy eating. We pilot tested the intervention in an elementary school on the Navajo Nation. Formative research revealed the importance of incorporating Diné culture, including traditional growing practices and the preparation of traditional foods into the curriculum. School staff also stressed the value of tying the curriculum to state and Diné educational standards. Students enjoyed opportunities for hands-on activities and snack preparation. Schools have a meaningful role to play in addressing childhood obesity disparities among Navajo children. School-based interventions that draw on cultural strengths and include healthy traditional practices can be a promising strategy for increasing fruit and vegetable consumption.


Aberrant expression of the neuronal transcription factor FOXP2 in neoplastic plasma cells.
FOXP2 mutation causes a severe inherited speech and language defect, while the related transcription factors FOXP1, FOXP3 and FOXP4 are implicated in cancer. FOXP2 mRNA and protein expression were characterised in normal human tissues, haematological cell lines and multiple myeloma (MM) patients' samples. FOXP2 mRNA and protein were absent in mononuclear cells from different anatomical sites, lineages and stages of differentiation. However, FOXP2 mRNA and protein was detected in several lymphoma (8/20) and all MM-derived cell lines (n = 4). FOXP2 mRNA was expressed in bone marrow samples from 96% of MM patients (24/25), 66.7% of patients with the pre-neoplastic plasma cell proliferation monoclonal gammopathy of undetermined significance (MGUS) (6/9), but not in reactive plasma cells. The frequency of FOXP2 protein expression in CD138(+) plasma cells was significantly higher in MGUS (P = 0.0005; mean 46.4%) and MM patients (P < or = 0.0001; mean 57.3%) than in reactive marrows (mean 2.5%). FOXP2 (>10% nuclear positivity) was detectable in 90.2% of MM (55/61) and 90.9% of MGUS (10/11) patients, showing more frequent expression than CD56 and labelling 75% of CD56-negative MM (9/12). FOXP2 represents the first transcription factor whose expression consistently differentiates normal and abnormal plasma cells and FOXP2 target genes are implicated in MM pathogenesis.


Prevalence of bronchial asthma and its associated factors among school children in urban Puducherry, India.
Bronchial asthma is an important public health problem at the global level. There is paucity of studies on bronchial asthma among school children in Urban India. To assess the prevalence and associated factors of bronchial asthma among school children. A cross-sectional study was conducted among 263 children studying in the 8(th), 9(th), and 10(th) standard using the modified International Study on Allergy and Asthma in Childhood questionnaire in Urban Puducherry. Data on associated factors that include family history of asthma, type of fuel used for cooking, placement of kitchen in the house, number of windows in sleeping room, pet animals, smoking among family members, birth order, and smoke outlet were collected. Data were analyzed by univariate analysis and expressed in proportion or percentages. Prevalence of ever bronchial asthma was found to be 5.3%, of which 4.2% had current episode of asthma during the last 1-year period. About 72.7% of the current asthmatics had cold or rhinitis and 54.5% each had itching or rashes and nocturnal dry cough. Prevalence is more among the 12-13 years age group (6.5%) compared to the 14-16 years age group (3.6%). Boys (5.4%) and girls (5.2%) had comparable prevalence rates. The prevalence was significantly more among those with a family history of asthma, having smoking habits in any of the family members, and the absence of smoke outlet in the house (P < 0.05). Bronchial asthma is an important health problem among children in urban Puducherry. Intervention on exposure to passive smoking and provision of smoke outlets may help to reduce the burden of disease at the community level.


The relationship between physical functional capacity and lung function in obese children and adolescents.
There is no consensus regarding obesity repercussions for lung function in children and adolescents. Therefore, the aim of the study was to determine whether obesity is associated with poor physical conditioning and damaged lung function in children and adolescents, and to correlate lung function with six-minute walk test (6MWT) results. This cross-sectional study included 38 obese subjects of both sexes, ranging between 5 and 17 years of age, as well as 56 control subjects paired by sex and age for the 6MWT, and 39 subjects for spirometry. Subjects performed spirometry according to the guidelines of the American Thoracic Society (ATS) and the European Respiratory Society. The obese group repeated spirometry after receiving bronchodilator (BD) treatments. Physical performance was evaluated via the 6MWT according to ATS guidelines. The obese group demonstrated lower forced expiratory volumes in the first second compared with the control group based on forced vital capacity indices (p < 0.01), findings consistent with airway obstruction in 36.8% of patients in the obese group. Walking distances were shorter in the obese group than in the control group. Changes in lung function did not correlate directly with performance on the 6MWT among obese patients. However, there was a correlation between lung function and variables indicative of effort during exercise. In the present study, the obese group walked shorter distances and demonstrated lower values in some markers of lung function. However, there is no relationship between their physical conditions and these test results. Therefore, we cannot conclusively state that poor physical performance results from damaged pulmonary function.


Distinct conformations of DNA-stabilized fluorescent silver nanoclusters revealed by electrophoretic mobility and diffusivity measurements.
Silver-DNA nanoclusters (Ag:DNAs) are novel fluorophores under active research and development as alternative biomolecular markers. Comprised of a few-atom Ag cluster that is stabilized in water by binding to a strand of DNA, they are also interesting for fundamental explorations into the properties of metal molecules. Here, we use in situ calibrated electrokinetic microfluidics and fluorescence correlation spectroscopy to determine the size, charge, and conformation of a select set of Ag:DNAs. Among them is a pair of spectrally distinct Ag:DNAs stabilized by the same DNA sequence, for which it is known that the silver cluster differs by two atoms. We find these two Ag:DNAs differ in size by ∼30%, even though their molecular weights differ by less than 3%. Thus a single DNA sequence can adopt very different conformations when binding slightly different Ag clusters. By comparing spectrally identical Ag:DNAs that differ in sequence, we show that the more compact conformation is insensitive to the native DNA secondary structure. These results demonstrate electrokinetic microfluidics as a practical tool for characterizing Ag:DNA.


Spousal physical violence against women during pregnancy.
To determine the prevalence of physical violence during pregnancy and the factors associated with it. A population-based, multicentre, cross sectional household survey. Rural, slum and urban non-slum areas of Bhopal, Chennai, Delhi, Lucknow, Nagpur, Trivandrum and Vellore, in India. A total of 9938 women who were 15 to 49 years of age and living with a child younger than 18 years old. Probability proportionate to size sampling of households was performed in three strata. Trained field workers administered a structured questionnaire. Women who reported domestic violence were asked about violence during pregnancy. Outcome variables included six violent behaviours: slap, hit, kick, beat, use of weapon and harm in any other way. Moderate to severe violence was defined as experience of any one or more of the following behaviours: hit, beat or kick. Odds ratios were calculated for risk and protective factors of violence during pregnancy using logistic regression. Physical spousal violence. The lifetime experience, during pregnancy, of being slapped was 16%, hit 10%, beat 10%, kicked 9%, use of weapon 5% and harmed in any other way 6%. Eighteen percent of women experienced at least one of these behaviours and 3% experienced all six. The overall prevalence of moderate to severe violence during pregnancy was 13%. Logistic regression showed that the factors determining whether a woman experienced moderate to severe violence during pregnancy were: husband accusing wife of an affair (OR 7.1; 95% CI 5.1 to 9.8), dowry harassment (OR 4.1; 95% CI 2.8 to 6.1), husband having an affair (OR 3.7; 95% CI 2.8 to 4.8), husband being regularly drunk (OR 3.2; 95% CI 2.6 to 4.1), low education of husband (OR 2.8; 95% CI 1.4 to 5.6), substance abuse by husband (OR 2.6; 95% CI 1.3 to 5.5), no social support (OR 1.8; 95% CI 1.1 to 3.0), three or more children (OR 1.6; 95% CI 1.2 to 2.1) and household crowding (OR 1.1; 95% CI 1.0 to 1.2). In this study, 12.9% of women experienced moderate to severe physical violence during pregnancy. Suspicion of infidelity, dowry harassment, husband being regularly drunk and low education of husband were the main risk factors for violence during pregnancy.


Antihypertensive treatment in renal transplant patients--is there a role for ACE inhibitors?
During the past two decades great progress was achieved with regards to short-term kidney graft survival. However, long-term graft survival did not improve similarly. Many factors contribute to chronic graft nephropathy eventually resulting in late graft loss, among these arterial hypertension is of major importance. In patients with chronic renal disease of diabetic and non-diabetic origin, angiotensin converting enzyme inhibitors have been convincingly shown to slow the progression of renal failure. The achieved nephroprotection correlates with the reduction of proteinuria by ACE inhibitor treatment. Also in renal transplant patients, ACE inhibitors have been shown unequivocally to reduce urinary protein excretion. The prevention of hyperfiltration, particular in the context of a reduced number of functional nephrons in patients with chronic graft nephropathy, could be important to prolong graft survival after renal transplantation. Moreover, ACE inhibitors may exert beneficial effects on immunologic processes contributing to chronic graft nephropathy. Many studies published in the last decade show convincingly that ACE inhibitors are safe and effective for the treatment of hypertension in renal allograft recipients. However, no data exist so far showing that ACE inhibitors are superior to other antihypertensive drugs in renal transplant patients and that they prolong graft survival. Studies investigating this issue are warranted. Apart from effects on the graft, ACE inhibitors may improve alterations of the cardiovascular system generally observed in renal transplant patients, such as structural alterations of large arteries, left ventricular hypertrophy, disturbed mechanical vessel wall properties and endothelial dysfunction. Therefore, angiotensin converting enzyme inhibitors could reduce cardiovascular morbidity and mortality in kidney transplant patients.


Perceptions of Primary Care Notes by Patients With Mental Health Diagnoses.
There are concerns regarding whether patients with mental illness should be provided with access to their electronic medical records. This study compared perceptions of patients with (n = 400) and without (n = 2,134) a mental health diagnosis regarding access to primary care clinic notes through secure online portals. Eligible participants viewed at least 1 clinic note during a 12-month period. Administrative data were used to stratify patients by mental health diagnosis. As we hypothesized, patients with and without mental health diagnoses had similar perceptions about online access to notes.


Host restriction of avian influenza viruses at the level of the ribonucleoproteins.
Although transmission of avian influenza viruses to mammals, particularly humans, has been repeatedly documented, adaptation and sustained transmission in the new host is a rare event that in the case of humans may result in pandemics. Host restriction involves multiple genetic determinants. Among the known determinants of host range, key determinants have been identified on the genes coding for the nucleoprotein and polymerase proteins that, together with the viral RNA segments, form the ribonucleoproteins (RNPs). The RNP genes form host-specific lineages and harbor host-associated genetic signatures. The functional significance of these determinants has been studied by reassortment and reverse genetics experiments, underlining the influence of the global genetic context. In some instances the molecular mechanisms have been approached, pointing to the importance of the polymerase activity and interaction with cellular host factors. Better knowledge of determinants of host restriction will allow monitoring of the pandemic potential of avian influenza viruses.


A logarithmic quantization index modulation for perceptually better data hiding.
In this paper, a novel arrangement for quantizer levels in the Quantization Index Modulation (QIM) method is proposed. Due to perceptual advantages of logarithmic quantization, and in order to solve the problems of a previous logarithmic quantization-based method, we used the compression function of mu-Law standard for quantization. In this regard, the host signal is first transformed into the logarithmic domain using the mu-Law compression function. Then, the transformed data is quantized uniformly and the result is transformed back to the original domain using the inverse function. The scalar method is then extended to vector quantization. For this, the magnitude of each host vector is quantized on the surface of hyperspheres which follow logarithmic radii. Optimum parameter mu for both scalar and vector cases is calculated according to the host signal distribution. Moreover, inclusion of a secret key in the proposed method, similar to the dither modulation in QIM, is introduced. Performance of the proposed method in both cases is analyzed and the analytical derivations are verified through extensive simulations on artificial signals. The method is also simulated on real images and its performance is compared with previous scalar and vector quantization-based methods. Results show that this method features stronger a watermark in comparison with conventional QIM and, as a result, has better performance while it does not suffer from the drawbacks of a previously proposed logarithmic quantization algorithm.


Characterization of a dominant cone degeneration in a green fluorescent protein-reporter mouse with disruption of Loci associated with human dominant retinal dystrophy.
PURPOSE. To characterize anatomically and functionally the retinal degeneration observed in a transgenic mouse line (OPN1LW-EGFP) expressing enhanced green fluorescent protein (EGFP) in a subpopulation of cone photoreceptors, and to map the location of the transgenic insertion. METHODS. An anatomic comparison of cone survival was carried out between wild type (WT) and transgenic mice at three postnatal time points (P80, P140, and P245). Retinal function was assessed at P245 by ERG and included an ultraviolet flicker stimulus to isolate S-cone function. Chromosomal mapping by FISH and high-resolution mapping on DNA fibers (Fiber-FISH) were performed to identify the location of the transgenic insertion. RESULTS. GFP expression was largely absent in S-cones. Cone numbers were significantly reduced in OPN1LW-EGFP mice at all time points compared to WT, with cone loss independent of GFP expression. Anatomic loss correlated with a functional deficit in dark- and light-adapted ERG responses, including a reduction in UV-flicker response, confirming the degeneration of S-cones. The phenotype of heterozygote mice was slightly less severe than in homozygotes, consistent with a dominantly inherited cone dystrophy. The transgenic insertion mapped to a specific region on chromosome 10 orthologous with loci for progressive bifocal chorioretinal atrophy and North Carolina macular dystrophy on human chromosome 6. CONCLUSIONS. Cone loss is global in OPN1LW-EGFP mice and is independent of GFP expression. The mechanism underlying the degeneration remains elusive; however, disruption of loci associated with dominantly inherited retinal degenerations in humans makes this mouse of great interest.


Progesterone resistance in endometriosis: link to failure to metabolize estradiol.
Endometriosis is the most common cause of pelvic pain and affects an estimated 5 million women in the US. The biologically active estrogen estradiol (E2) is the best-defined mitogen for the growth and inflammation processes in the ectopic endometriotic tissue that commonly resides on the pelvic organs. Progesterone and progestins may relieve pain by limiting growth and inflammation in endometriosis but a portion of patients with endometriosis and pelvic pain do not respond to treatment with progestins. Moreover, progesterone-induced molecular changes in the eutopic (intrauterine) endometrial tissue of women with endometriosis are either blunted or undetectable. These in vivo observations are indicative of resistance to progesterone action in endometriosis. The molecular basis of progesterone resistance in endometriosis may be related to an overall reduction in the levels of progesterone receptors (PRs) and the lack of the PR isoform named progesterone receptor B (PR-B). In normal endometrium, progesterone acts on stromal cells to induce secretion of paracrine factor(s). These unknown factor(s) act on neighboring epithelial cells to induce the expression of the enzyme 17beta-hydroxysteroid dehydrogenase type 2 (17beta-HSD-2), which metabolizes the biologically active estrogen E2 to estrone (E1). In endometriotic tissue, progesterone does not induce epithelial 17beta-HSD-2 expression due to a defect in stromal cells. The inability of endometriotic stromal cells to produce progesterone-induced paracrine factors that stimulate 17beta-HSD-2 may be due to the lack of PR-B and very low levels of progesterone receptor A (PR-A) observed in vivo in endometriotic tissue. The end result is deficient metabolism of E2 in endometriosis giving rise to high local concentrations of this local mitogen. The cellular and molecular mechanisms underlying progesterone resistance and failure to metabolize E2 in endometriosis are reviewed.


Probing single molecule dynamics.
The room temperature dynamics of single sulforhodamine 101 molecules dispersed on a glass surface are investigated on two different time scales with near-field optics. On the 10(-2)- to 10(2)-second time scale, intensity fluctuations in the emission from single molecules are examined with polarization measurements, providing insight into their spectroscopic properties. On the nanosecond time scale, the fluorescence lifetimes of single molecules are measured, and their excited-state energy transfer to the aluminum coating of the near-field probe is characterized. A movie of the time-resolved emission demonstrates the feasibility of fluorescence lifetime imaging with single molecule sensitivity, picosecond temporal resolution, and a spatial resolving power beyond the diffraction limit.


Cervical vertebrae anomalies in subjects with Class II malocclusion assessed by lateral cephalogram and cone beam computed tomography.
A high prevalence of cervical vertebrae anomalies (CVA) has been recently associated with various malocclusions. Our aim was to study the prevalence of CVA on lateral cephalograms in Class II subjects and to compare the findings with those obtained from cone beam computed tomography (CBCT). Standardized cephalograms of 238 Class II patients were analysed for CVA. Cephalogram and CBCT were available for an additional 21 subjects. Cephalometric values were correlated with vertebrae morphology; logistic regressions and intraobserver agreement were evaluated. Inspection of lateral cephalograms could exclude CVA in 90.3 per cent of the subjects, while 9.7 per cent showed potential fusions. No correlations were found between the cephalometric values and potential vertebrae anomalies. In the 21 patients with a CBCT and a lateral cephalogram, the visual assessment of the cephalogram yielded a potential fusion in nine cases. None could be confirmed by CBCT. A low number of potentially fused cervical vertebrae could be detected on lateral cephalograms. The possible fusions did not correlate to any cephalometric values nor could they be confirmed by CBCT, the gold standard for assessing CVA. Visual examination of a cephalogram may result in a false-positive finding and does not allow reliable diagnosis of CVA.


Whole-Proteome Peptide Microarrays for Profiling Autoantibody Repertoires within Multiple Sclerosis and Narcolepsy.
The underlying molecular mechanisms of autoimmune diseases are poorly understood. To unravel the autoimmune processes across diseases, comprehensive and unbiased analyses of proteins targets recognized by the adaptive immune system are needed. Here we present an approach starting from high-density peptide arrays to characterize autoantibody repertoires and to identify new autoantigens. A set of ten plasma and serum samples from subjects with multiple sclerosis, narcolepsy, and without any disease diagnosis were profiled on a peptide array representing the whole proteome, hosting 2.2 million 12-mer peptides with a six amino acid lateral shift. On the basis of the IgG reactivities found on these whole-proteome peptide microarrays, a set of 23 samples was then studied on a targeted array with 174 000 12-mer peptides of single amino acid lateral shift. Finally, verification of IgG reactivities was conducted with a larger sample set (n = 448) using the bead-based peptide microarrays. The presented workflow employed three different peptide microarray formats to discover and resolve the epitopes of human autoantibodies and revealed two potentially new autoantigens: MAP3K7 in multiple sclerosis and NRXN1 in narcolepsy. The presented strategy provides insights into antibody repertoire reactivity at a peptide level and may accelerate the discovery and validation of autoantigens in human diseases.


Geometrical and energetical structural changes in organic dyes for dye-sensitized solar cells probed using photoelectron spectroscopy and DFT.
The effects of alkoxy chain length in triarylamine based donor-acceptor organic dyes are investigated with respect to the electronic and molecular surface structures on the performance of solar cells and the electron lifetime. The dyes were investigated when adsorbed on TiO2 in a configuration that can be used for dye-sensitized solar cells (DSCs). Specifically, the two dyes D35 and D45 were compared using photoelectron spectroscopy (PES) and density functional theory (DFT) calculations. The differences in solar cell characteristics when longer alkoxy chains are introduced in the dye donor unit are attributed to geometrical changes in dye packing while only minor differences were observed in the electronic structure. A higher dye load was observed for D45 on TiO2. However, D35 based solar cells result in higher photocurrent although the dye load is lower. This is explained by different geometrical structures of the dyes on the surface.


An overview of American higher education.
This overview of postsecondary education in the United States reviews the dramatic changes over the past fifty years in the students who go to college, the institutions that produce higher education, and the ways it is financed. The article, by Sandy Baum, Charles Kurose, and Michael McPherson, creates the context for the articles that follow on timely issues facing the higher education community and policy makers. The authors begin by observing that even the meaning of college has changed. The term that once referred primarily to a four-year period of academic study now applies to virtually any postsecondary study--academic or occupational, public or private, two-year or four-year-- that can result in a certificate or degree. They survey the factors underlying the expansion of postsecondary school enrollments; the substantial increases in female, minority, disadvantaged, and older students; the development of public community colleges; and the rise of for-profit colleges. They discuss the changing ways in which federal and state governments help students and schools defray the costs of higher education as well as more recent budget tensions that are now reducing state support to public colleges. And they review the forces that have contributed to the costs of producing higher education and thus rising tuitions. The authors also cite evidence on broad measures of college persistence and outcomes, including low completion rates at community and for-profit colleges, the increasing need for remedial education for poorly prepared high school students, and a growing gap between the earnings of those with a bachelor's degree and those with less education. They disagree with critics who say that investments in higher education, particularly for students at the margin, no longer pay off. A sustained investment in effective education at all levels is vital to the nation's future, they argue. But they caution that the American public no longer seems willing to pay more for more students to get more education. They therefore urge the higher education community to make every effort to find innovations, including creative uses of information technology, that can hold down costs while producing quality education.


Application of information theory to decision analysis in potentially prostaglandin-responsive neonates.
To define settings in which use of prostaglandin E1 before transfer from a community hospital to a tertiary care center benefits neonates with possible heart disease, information theory was used to predict the probability of a favorable response to prostaglandin therapy from the limited information of clinical variables. Records of 250 patients, newborn to 7 days old, with suspected heart disease were reviewed to assess six clinical variables (cyanosis, respiratory distress, heart murmur, pulse contour, hepatomegaly and prematurity). According to the anatomic and hemodynamic cardiovascular condition, each case was categorized as to whether a favorable response to prostaglandin E1 could be anticipated. Information content of each clinical variable with respect to prostaglandin responsiveness was determined, and patients were classified according to the most informative clinical variable. Stepwise extraction of information proceeded until remaining clinical variables added no significant information. Bayes' rule gave estimates of probability of prostaglandin-responsive defect in final subgroups for use in decision analysis. Cyanosis, murmur, small volume pulses and prematurity gave information about prostaglandin-responsive defects. Decision analysis indicated that frequency of poor outcome is minimized by early prostaglandin treatment of cyanotic term infants with a murmur or poor pulses, regardless of how ill they appear, and by treating any critically ill term newborn who has either cyanosis or poor pulses. Acyanotic patients with normal pulses are best untreated with prostaglandin until after definitive diagnosis is made. Advantage to either course was not seen in some small subgroups. Information theory with decision analysis is a rigorous approach to identify relevant clinical variables and define their roles in critical decisions in pediatric cardiology.


Management of persistent pain in the older patient: a clinical review.
Persistent pain is highly prevalent, costly, and frequently disabling in later life. To describe barriers to the management of persistent pain among older adults, summarize current management approaches, including pharmacologic and nonpharmacologic modalities; present rehabilitative approaches; and highlight aspects of the patient-physician relationship that can help to improve treatment outcomes. This review is relevant for physicians who seek an age-appropriate approach to delivering pain care for the older adult. Search of MEDLINE and the Cochrane database from January 1990 through May 2014, using the search terms older adults, senior, ages 65 and above, elderly, and aged along with non-cancer pain, chronic pain, persistent pain, pain management, intractable pain, and refractory pain to identify English-language peer-reviewed systematic reviews, meta-analyses, Cochrane reviews, consensus statements, and guidelines relevant to the management of persistent pain in older adults. Of the 92 identified studies, 35 evaluated pharmacologic interventions, whereas 57 examined nonpharmacologic modalities; the majority (n = 50) focused on older adults with osteoarthritis. This evidence base supports a stepwise approach with acetaminophen as first-line therapy. If treatment goals are not met, a trial of a topical nonsteroidal anti-inflammatory drug, tramadol, or both is recommended. Oral nonsteroidal anti-inflammatory drugs are not recommended for long-term use. Careful surveillance to monitor for toxicity and efficacy is critical, given that advancing age increases risk for adverse effects. A multimodal approach is strongly recommended-emphasizing a combination of both pharmacologic and nonpharmacologic treatments to include physical and occupational rehabilitation, as well as cognitive-behavioral and movement-based interventions. An integrated pain management approach is ideally achieved by cultivating a strong therapeutic alliance between the older patient and the physician. Treatment planning for persistent pain in later life requires a clear understanding of the patient's treatment goals and expectations, comorbidities, and cognitive and functional status, as well as coordinating community resources and family support when available. A combination of pharmacologic, nonpharmacologic, and rehabilitative approaches in addition to a strong therapeutic alliance between the patient and physician is essential in setting, adjusting, and achieving realistic goals of therapy.


Multilocus sequence typing (MLST) and M13 PCR fingerprinting revealed heterogeneity amongst Cryptococcus species obtained from Italian veterinary isolates.
Cryptococcosis represents a fungal disease acquired from the environment with animals serving as host sentinels for human exposure. The aim of this study was to investigate the genetic characteristics of Cryptococcus isolates from veterinary sources (cats, dogs and birds) to understand their epidemiology and the genetic variability of the casual isolates. Mating-type PCR in connection with MLST analysis using the ISHAM consensus MLST scheme for the C. neoformans/C. gattii species complex was used to genotype 17 C. neoformans isolates. In the absence of an MLST typing scheme Cryptococcus adeliensis, C. albidus, C. aureus, C. carnescens, C. laurentii, C. magnus and C. uniguttulatus strains were typed using M13 PCR fingerprinting. All C. neoformans isolates were MATα mating type, but hybrids possessed αADa and aADα mating and serotypes. Two C. neoformans molecular types VNI, VNIV and VNIII and VNII/VNIV hybrids were identified. Amongst the 66 non-C. neoformans strains investigated 55 M13 PCR fingerprinting types were identified. The wide variety of MLST types of C. neoformans and the occurrence of αADa and aADα hybrids in our study supports the notion of genetic recombination in the area studied. The heterogeneity of the non-C. neoformans isolates remains open to further investigations and should be taken into consideration when identifying emergent pathogens.


Intracoronal sealing ability of two dental cements.
The purpose of this study was to compare the efficacy of sealing the coronal 2-mm of the root canals versus covering the entire pulpal floor with one of two dental-resin cements (Principle or C&B Metabond). Sixty-two molars with the occlusal half of the crowns and the apical half of the roots removed were used. Each canal was enlarged by using a #3 Gates Glidden bur and obturated with unsealed gutta-percha cones. The teeth were randomly assigned to four groups, each containing 15 teeth, plus a negative and a positive control. In group 1, 2 mm of Principle were placed over the entire pulpal floor. In group 2, Principle was placed 2 mm into each canal orifice. Groups 3 and 4 were the same as groups 1 and 2, except C&B Metabond cement was used. After the cement set, the gutta-percha was removed and the integrity of the seal was tested by fluid filtration at a pressure of 20 cm H2O at 1 h and at 1, 2, and 4 weeks. The data were analyzed by a three-way ANOVA and the Student-Newman-Keuls tests at alpha = 0.05. The controls behaved as expected. Results showed that there were no statistically significant differences among the materials used or the location (p > 0.05), but there was a significant difference with respect to time. Principle leaked significantly more than C&B Metabond at 1 h (p < 0.05), but the seal became tighter over time. C&B Metabond leaked less early (p < 0.05) but increased in leakage at 4 weeks. Both materials sealed well over the 4-week study. Principle was easier to use, and sealing the entire pulpal floor was easier than sealing only the canal orifice.


The Memorandum of Good Practice: theory versus application.
The goal of this study was to evaluate the quality of investigative interviews in England and Wales since implementation of the Memorandum of Good Practice (MOGP), which specified how forensic interviews of alleged child abuse victims should be conducted. Transcripts of 119 videotaped interviews of alleged victims between the ages of 4 and 13 years were obtained from 13 collaborating police forces. Trained raters then classified the types of prompts used by the investigators to elicit substantive information from the children, and tabulated the number of forensically relevant details provided by the children in each response. Like their counterparts in the United States, Israel, and Sweden, forensic interviewers in England and Wales relied heavily on option-posing prompts, seldom using open-ended utterances to elicit information from the children. Nearly 40% of the information obtained was elicited using option-posing and suggestive prompts, which are known to elicit less reliable information than open-ended prompts do. Despite the clarity and specificity of the MOGP, its implementation appears to have had less effect on the practices of forensic interviewers in the field than was hoped. Further work should focus on ways of training interviewers to implement the superior practices endorsed by the MOGP and similar professional guidelines.


Distribution of the monocarboxylate transporter MCT2 in human cerebral cortex: an immunohistochemical study.
The monocarboxylate transporter MCT2 belongs to a large family of membrane proteins involved in the transport of lactate, pyruvate and ketone bodies. Although its expression in rodent brain has been well documented, the presence of MCT2 in the human brain has been questioned on the basis of low mRNA abundance. In this study, the distribution of the monocarboxylate transporter MCT2 has been investigated in the cortex of normal adult human brain using an immunohistochemical approach. Widespread neuropil staining in all cortical layers was observed by light microscopy. Such a distribution was very similar in three different cortical areas investigated. At the cellular level, the expression of MCT2 could be observed in a large number of neurons, in fibers both in grey and white matter, as well as in some astrocytes, mostly localized in layer I and in the white matter. Double staining experiments combined with confocal microscopy confirmed the neuronal expression but also suggested a preferential postsynaptic localization of synaptic MCT2 expression. A few astrocytes in the grey matter appeared to exhibit MCT2 labelling but at low levels. Electron microscopy revealed strong MCT2 expression at asymmetric synapses in the postsynaptic density and also within the spine head but not in the presynaptic terminal. These data not only demonstrate neuronal MCT2 expression in human, but since a portion of it exhibits a distinct synaptic localization, it further supports a putative role for MCT2 in adjustment of energy supply to levels of activity.


Redox sensing by a Rex-family repressor is involved in the regulation of anaerobic gene expression in Staphylococcus aureus.
An alignment of upstream regions of anaerobically induced genes in Staphylococcus aureus revealed the presence of an inverted repeat, corresponding to Rex binding sites in Streptomyces coelicolor. Gel shift experiments of selected upstream regions demonstrated that the redox-sensing regulator Rex of S. aureus binds to this inverted repeat. The binding sequence--TTGTGAAW(4)TTCACAA--is highly conserved in S. aureus. Rex binding to this sequence leads to the repression of genes located downstream. The binding activity of Rex is enhanced by NAD+ while NADH, which competes with NAD+ for Rex binding, decreases the activity of Rex. The impact of Rex on global protein synthesis and on the activity of fermentation pathways under aerobic and anaerobic conditions was analysed by using a rex-deficient strain. A direct regulatory effect of Rex on the expression of pathways that lead to anaerobic NAD+ regeneration, such as lactate, formate and ethanol formation, nitrate respiration, and ATP synthesis, is verified. Rex can be considered a central regulator of anaerobic metabolism in S. aureus. Since the activity of lactate dehydrogenase enables S. aureus to resist NO stress and thus the innate immune response, our data suggest that deactivation of Rex is a prerequisite for this phenomenon.


Deciphering Resistome and Virulome Diversity in a Porcine Slaughterhouse and Pork Products Through Its Production Chain.
We aimed to better understand resistome and virulome patterns on animal and process-area surfaces through a pig slaughterhouse to track possible contamination within the food production chain. Culture-dependent methods revealed high levels of microbial contamination, corresponding to mesophilic and pathogenic bacteria on both the animal and process-area surfaces mainly in the anesthesia (AA and AS) zone followed by "scorching and whip" (FA and FS) zone and also in the end products. To evaluate the potential risk of antibiotic resistance and virulence determinants, shotgun metagenomic DNA-sequencing of isolates from selected areas/products uncovered a high diversity and richness of antibiotic resistance genes (ARGs): 55-62 genes in the anesthesia area (AA and AS) and 35-40 in "animal-arrival zone" (MA and MS). The "scorching and whip" (FA and FS) area, however, exhibited lowered abundance of ARGs (1-6), indicating that the scalding and depilating process (an intermediate zone between "anesthesia" and "scorching and whip") significantly decreased bacterial load by 1-3 log10 but also diminished the resistome. The high prevalence of antibiotic-inactivating enzyme genes in the "animal-arrival zone" (60-65%) and "anesthesia" area (56%) were mainly represented by those for aminoglycoside (46-51%) and lincosamide (14-19%) resistance, which did not reflect selective pressures by antibiotics most commonly used in pig therapy-tetracyclines and beta-lactams. Contrary to ARGs, greater number of virulence resistance genes were detected after evisceration in some products such as kidney, which reflected the poor hygienic practices. More than 19 general virulence features-mainly adherence, secretion system, chemotaxis and motility, invasion and motility were detected in some products. However, immune evasion determinants were detected in almost all samples analyzed from the beginning of the process, with highest amounts found from the anesthesia area. We conclude that there are two main sources of contamination in a pig slaughterhouse: the microorganisms carried on the animals' hide, and those from the evisceration step. As such, focussing control measures, e.g., enhanced disinfection procedures, on these contamination-source areas may reduce risks to food safety and consumer health, since the antibiotic and virulence determinants may spread to end products and the environment; further, ARG and virulence traits can exacerbate pathogen treatments.


3T1R model and tuning of thermoluminescence intensity by optimization of dopant concentration in monoclinic Gd2O3:Er3+;Yb3+ co-doped phosphor.
Thermoluminescence property of a phosphor is an important parameter that helps to determine the use of a phosphor in various dosimetric applications. In this study, the thermoluminescence behaviour of a monoclinic Gd2O3:Er3+;Yb3+ nanophosphor was studied. Gd2O3:Er3+;Yb3+ nanophosphor was prepared using a combustion synthesis method. Structural characterization was carried out via X-ray diffraction and electron microscopy methods. Herein, thermoluminescence (TL) study and kinetic analysis of the UV- and gamma-irradiated phosphor was also carried out. The prepared phosphor exhibits two TL glow curves around 122 and 263 °C for UV excitation and 157 and 295 °C for gamma exposure. The effect of different parameters on the TL glow curve was investigated. The glow curve deconvolution function was applied on the tuned glow peak curve. The trapping parameters were determined for tuned glow curve peaks as well as for the deconvoluted peaks. Moreover, reproducibility of the sample was determined via 7 replicate TL measurements. The prepared sample shows good reproducibility for both UV and gamma exposure. A theoretical three trap and one recombination centre (3T1R) model was proposed to explain the concentration quenching effect on the thermoluminescence behaviour of the prepared sample.


Polychlorinated biphenyls in urban lake sediments from wuhan, central China: occurrence, composition, and sedimentary record.
Nine surface sediments and a dated sediment core collected from urban lakes in Wuhan, Central China, were analyzed to investigate the concentrations, occurrence, composition, and depositional fluxes of polychlorinated biphenyls (PCBs). The concentrations of SigmaPCB (the sum of 39 congeners) in surface sediments ranged from 0.90 to 46.14 ng g(-1) dry weight. Only in Longyang Lake and Nantaizi Lake did concentrations of SigmaPCB exceed the effects range low value. The concentrations of SigmaPCB in the sediment core varied from 1.3 to 43.1 ng g(-1). The profile of SigmaPCB concentrations closely reflected the changes in production and usage of PCBs in this region, and the profiles of SigmaPCB fluxes were similar to those of SigmaPCB concentrations, except for a distinct decrease in SigmaPCB fluxes after about 1998. This was possibly due to more large-scale cases of land use and increasingly rapidly urban development occurring in China since 2000, resulting in increased lake sediment fluxes but reduced SigmaPCB fluxes. The results suggest that urban run-off and wet deposition leaching PCBs off the land and into the lake may be the most important source of PCBs in Donghu Lake. Sedimentary profiles for PCB congeners showed a decrease in concentrations in the following order: penta approximately hexa- > tetra- > tri- > hepta-PCBs. The relative abundances of tri- and tetra-PCBs in the core accounted for more than 80% of the total PCBs detected in sediments deposited before their first commercial use, suggesting the post-depositional mobilization of less chlorinated PCB congeners in the sediment core. This is the first study to estimate the spatial and historical trends of PCBs in subtropical urban lakes.


The Effects of Elaboration on the Strength of Doping-Related Attitudes: Resistance to Change and Behavioral Intentions.
This experiment analyzed whether attitudes toward the legalization of several doping behaviors would resist change and predict behavioral intentions when they were initially formed through thoughtful (i.e., high elaboration) versus nonthoughtful (i.e., low elaboration) processes. Participants were randomly assigned first to a persuasive message either against or in favor of the legalization, which they read with relatively high or low degrees of deliberative thinking. Attitudes and intentions regarding legalization were assessed following that message. Next, each participant received a second message that was opposed to the first one, serving as an attack against the attitude that participants had just formed. Finally, attitudes were again assessed. As hypothesized, participants showed greater attitude-consistent intentions when they formed their initial attitudes through thoughtful (vs. nonthoughtful) consideration of the first message. Moreover, the second message resulted in greater resistance to attitude change when participants formed their initial attitudes through thoughtful (vs. nonthoughtful) processes.


Efficacy of amisulpride for depressive symptoms in individuals with mental disorders: A systematic review and meta-analysis.
Depressive symptoms occur in several psychiatric disorders, often in the absence of a formal diagnosis of depression. We aimed to evaluate the efficacy and the tolerability of amisulpride, both alone and as augmentation therapy, in the treatment of depressive symptoms in individuals with any major psychiatric disorder. We searched PubMed, Embase, PsycINFO, GreyLit, OpenGrey and ProQuest up to March 2020 for randomised controlled trials focussing on the treatment of an acute depressive episode in any major psychiatric disorder. A random-effect meta-analysis was performed to synthesize the findings on depressive symptoms (primary outcome), response rate and tolerability. We retrieved 11 studies including 2065 patients with a diagnosis of dysthymia (eight studies), major depression (one study) or schizophrenia (two studies). Amisulpride 50 mg/day was associated with a larger reduction of depressive symptoms compared to placebo (standardised mean difference [SMD] = -0.70, CI 95% -0.92, -0.49; I2 = 0.0%), and was found to be comparable to selective serotonin reuptake inhibitors (SSRIs; SMD = -0.08, CI 95% -0.23, 0.06, I2 = 0.0%), amineptine, imipramine and amitriptyline in the treatment of dysthymia (three studies, not pooled). In individuals with schizophrenia, amisulpride administered at higher doses (>400 mg/day) was comparable to olanzapine and risperidone (two studies, not pooled). In terms of tolerability, amisulpride was superior to placebo for dysthymia (odds ratio [OR] = 3.94, CI 95% 1.07, 14.48; I2 = 0.0) and comparable with SSRIs (OR = 0.94, CI 95% 0.55, 1.62; I2 = 0.0%). Treatment with amisulpride could be a valid choice for selected individuals with dysthymia or depressive symptoms in the context of schizophrenia. More studies on the efficacy and tolerability of amisulpride are needed to draw firm conclusions on its potential benefits in other psychiatric disorders.


Attitude May Be Everything, But Is Everything an Attitude? Cognitive Distortions May Not Be Evaluations of Rape.
Cognitive distortions are often referred to as attitudes toward rape in theory, research, and clinical practice pertaining to sexual aggression. In the social-psychological literature, however, attitudes are typically defined as evaluations; thus, in this context, attitudes toward rape are considered evaluations of rape (e.g., rape is negative vs. positive). The purpose of the current study was to explore whether a widely used measure of cognitive distortions (RAPE Scale; Bumby, 1996) assesses evaluation of rape, and, if not, whether evaluation of rape and the cognitions assessed by the RAPE Scale are independently associated with sexually aggressive behavior. Participants (660 male undergraduate students) completed the RAPE Scale as well as measures of evaluation of rape and sexually aggressive behavior. An exploratory factor analysis revealed that the RAPE Scale items formed a correlated but distinct factor from the Evaluation of Rape Scale items. Regression analyses indicated that the Evaluation of Rape Scale and the RAPE Scale had small to moderate independent associations with self-report measures of sexually aggressive behavior. Our results suggest that evaluation of rape may be distinct from cognitive distortions regarding rape, and both evaluation and cognitive distortions may be relevant for understanding sexual violence.


Serum concentrations of polychlorinated dibenzo-p-dioxins among ceramicists.
Polychlorinated dibenzo-p-dioxins (PCDDs) occur naturally in ball clay at elevated concentrations. Thus, persons who habitually work with clay may be at risk for exposure to PCDDs. An earlier case report provided some evidence of elevated PCDD levels in serum for long-term hobby ceramicists; however, no previous study has measured serum dioxin concentrations among ceramicists. This study measured PCDD serum levels for 27 individuals involved in ceramics making. The average residual, defined as the average of the [log measured serum lipid concentration – log background serum lipid concentration], was calculated and then tested to determine whether it was significantly different from zero. The p-values for the average residuals indicated that the serum lipid concentrations for several PCDD congeners were elevated relative to background. The number of significant residuals increased dramatically if the background concentrations were adjusted to account for the fact that they were not contemporaneous with the measurements for the ceramicists. The ratio of the 1,2,3,6,7,8-HxCDD concentration to the 1,2,3,7,8,9-HxCDD concentration was greater than 1.0, unlike in ball clay, suggesting that although long-term working with ball clay elevates the PCDD levels in serum somewhat, it is not the predominant source of the PCDD body burden for ceramicists.


Evaluating Nurses Acceptance of Hospital Information Systems: A Case Study of a Tertiary Care Hospital.
This study aims at evaluating hospital information systems (HIS) acceptance factors among nurses, in order to provide suggestions for successful HIS implementation. The study used mainly quantitative survey methods to collect data directly from nurses through a questionnaire. The availability of computers in the hospital was one of the most influential factors, with a special emphasis on the unavailability of laptop computers and computers on wheels to facilitate immediate data entry and retrieval when nurses are at the point of care. Nurses believed that HIS might frequently slow down the process of care delivery and increase the time spent by patients inside the hospital especially during slow performance and responsiveness phases. Recommendations were classified into three main areas; improving system performance and availability of computers in the hospital, increasing organizational support in the form of providing training and protected time for nurses' to learn and enhancing users' feedback by listening to their complaints and considering their suggestions.


Effect of different treatment regimen with dexamethasone and acetylcysteine on corneal wound healing in rabbits.
The effect of corticosteroid therapy on corneal wound healing is controversial. The objective of this study was to evaluate the effects of combination therapy with dexamethasone and acetylcysteine at different times and durations on experimentally-induced corneal wounds and haze in rabbits. Eighteen adult New Zealand white rabbits were divided into three groups of six each. Under anesthesia corneal wounds were created surgically in the center of all eyes. The right eyes of rabbits in group 1 were treated topically with acetylcysteine and dexamethasone immediately after surgery, those in group 2 were treated with acetylcysteine from day 1 and with acetylcysteine and dexamethasone from day 8, and those in group 3 were treated with acetylcysteine from day 1 and with acetylcysteine and dexamethasone from day 15. The left eyes were assigned as controls and were treated with normal saline. All eyes were treated six times a day for 28 days. Corneal wounds were measured by fluorescein staining every day. The combination of acetylcysteine and dexamethasone in group 1 significantly increased mean healing time, but did not change that in groups 2 and 3. Clinical and histopathologic examinations revealed that one month after the ulceration in groups 1 corneal haze was greater in treated than in the control eyes. Moreover, there was no significant difference between the control and treated eyes of group 1, 2, or 3 in terms of corneal haze at two or three months after the ulceration. The findings of the present study show that the association of 3% concentration of NAC and 0.1% concentration of dexamethasone immediately after corneal ulceration can delay corneal wound healing, and consequently produce more corneal haze. Thus, the use of 0.1% concentration of dexamethasone should be delayed at least until the completion of the epithelial defects.


De novo SCN2A splice site mutation in a boy with Autism spectrum disorder.
SCN2A is a gene that codes for the alpha subunit of voltage-gated, type II sodium channels, and is highly expressed in the brain. Sodium channel disruptions, such as mutations in SCN2A, may play an important role in psychiatric disorders. Recently, de novo SCN2A mutations in autism spectrum disorder (ASD) have been identified. The current study characterizes a de novo splice site mutation in SCN2A that alters mRNA and protein products. We describe results from clinical and genetic characterizations of a seven-year-old boy with ASD. Psychiatric interview and gold standard autism diagnostic instruments (ADOS and ADI-R) were used to confirm ASD diagnosis, in addition to performing standardized cognitive and adaptive functioning assessments (Leiter-R and Vineland Adaptive Behavior Scale), and sensory reactivity assessments (Sensory Profile and Sensory Processing Scales). Genetic testing by whole exome sequencing revealed four de novo events, including a splice site mutation c.476 + 1G > A in SCN2A, a missense mutation (c.2263G > A) causing a p.V755I change in the TLE1 gene, and two synonymous mutations (c.2943A > G in the BUB1 gene, and c.1254 T > A in C10orf68 gene). The de novo SCN2A splice site mutation produced a stop codon 10 amino acids downstream, possibly resulting in a truncated protein and/or a nonsense-mediated mRNA decay. The participant met new DSM-5 criteria for ASD, presenting with social and communication impairment, repetitive behaviors, and sensory reactivity issues. The participant's adaptive and cognitive skills fell in the low range of functioning. This report indicates that a splice site mutation in SCN2A might be contributing to the risk of ASD. Describing the specific phenotype associated with SCN2A mutations might help to reduce heterogeneity seen in ASD.


Psychological Health, Well-Being, and the Mind-Heart-Body Connection: A Scientific Statement From the American Heart Association.
As clinicians delivering health care, we are very good at treating disease but often not as good at treating the person. The focus of our attention has been on the specific physical condition rather than the patient as a whole. Less attention has been given to psychological health and how that can contribute to physical health and disease. However, there is now an increasing appreciation of how psychological health can contribute not only in a negative way to cardiovascular disease (CVD) but also in a positive way to better cardiovascular health and reduced cardiovascular risk. This American Heart Association scientific statement was commissioned to evaluate, synthesize, and summarize for the health care community knowledge to date on the relationship between psychological health and cardiovascular health and disease and to suggest simple steps to screen for, and ultimately improve, the psychological health of patients with and at risk for CVD. Based on current study data, the following statements can be made: There are good data showing clear associations between psychological health and CVD and risk; there is increasing evidence that psychological health may be causally linked to biological processes and behaviors that contribute to and cause CVD; the preponderance of data suggest that interventions to improve psychological health can have a beneficial impact on cardiovascular health; simple screening measures can be used by health care providers for patients with or at risk for CVD to assess psychological health status; and consideration of psychological health is advisable in the evaluation and management of patients with or at risk for CVD.


Causes of urinary incontinence after acute hemispheric stroke.
We prospectively studied bladder function in stroke patients to determine the mechanisms responsible for poststroke urinary incontinence. Fifty-one patients with recent unilateral ischemic hemispheric stroke admitted to a neurorehabilitation unit were enrolled. The presence of urinary incontinence was correlated with infarct location, neurological deficits, and functional status. Urodynamic studies were performed on all incontinent patients. Nineteen patients (37%) were incontinent. Incontinence was associated with large infarcts, aphasia, cognitive impairment, and functional disability (p < 0.05) but not with age, sex, side of stroke, or time from stroke to entry in the study. Urodynamic studies, performed on all 19 incontinent patients, revealed bladder hyperreflexia in 37%, normal studies in 37%, bladder hyporeflexia in 21%, and detrusor-sphincter dyssynergia in 5%. All of the patients with normal urodynamic studies were aphasic, demented, or severely functionally impaired. All of the patients with hyporeflexic bladders had underlying diabetes or were taking anticholinergic medications. Forty-six percent of incontinent patients treated with scheduled voiding alone were continent at discharge compared with 17% of patients treated pharmacologically. There are three major mechanisms responsible for poststroke urinary incontinence: 1) disruption of the neuromicturition pathways, resulting in bladder hyperreflexia and urgency incontinence; 2) incontinence due to stroke-related cognitive and language deficits, with normal bladder function; and 3) concurrent neuropathy or medication use, resulting in bladder hyporeflexia and overflow incontinence. Urodynamic studies are of benefit in establishing the cause of incontinence. Scheduled voiding is a useful first-line treatment in many cases of incontinence.


An overview of deep learning in medical imaging focusing on MRI.
What has happened in machine learning lately, and what does it mean for the future of medical image analysis? Machine learning has witnessed a tremendous amount of attention over the last few years. The current boom started around 2009 when so-called deep artificial neural networks began outperforming other established models on a number of important benchmarks. Deep neural networks are now the state-of-the-art machine learning models across a variety of areas, from image analysis to natural language processing, and widely deployed in academia and industry. These developments have a huge potential for medical imaging technology, medical data analysis, medical diagnostics and healthcare in general, slowly being realized. We provide a short overview of recent advances and some associated challenges in machine learning applied to medical image processing and image analysis. As this has become a very broad and fast expanding field we will not survey the entire landscape of applications, but put particular focus on deep learning in MRI. Our aim is threefold: (i) give a brief introduction to deep learning with pointers to core references; (ii) indicate how deep learning has been applied to the entire MRI processing chain, from acquisition to image retrieval, from segmentation to disease prediction; (iii) provide a starting point for people interested in experimenting and perhaps contributing to the field of deep learning for medical imaging by pointing out good educational resources, state-of-the-art open-source code, and interesting sources of data and problems related medical imaging.


The mechanosensory-motor apparatus of antennae in the Oleander hawk moth (Daphnis nerii, Lepidoptera).
Insect antennae are sensory organs of great importance because they can sense diverse environmental stimuli. In addition to serving as primary olfactory organs of insects, antennae also sense a wide variety of mechanosensory stimuli, ranging from low-frequency airflow or gravity cues to high-frequency antennal vibrations due to sound, flight or touch. The basal segments of the antennae house multiple types of mechanosensory structures that prominently include the sensory hair plates, or Böhm's bristles, which measure the gross extent of antennal movement, and a ring of highly sensitive scolopidial neurons, collectively called the Johnston's organs, which record subtle flagellar vibrations. To fulfill their multifunctional mechanosensory role, the antennae of insects must actively move thereby enhancing their ability to sense various cues in the surrounding environment. This tight coupling between antennal mechanosensory function and antennal movements means that the underlying mechanosensory-motor apparatus constitutes a highly tuned feedback-controlled system. Our study aims to explore how the sensory and motor components of this system are configured to enable such functional versatility. We describe antennal mechanosensory neurons, their central projections in the brain relative to antennal motor neurons and the internal morphology of various antennal muscles that actuate the basal segments of the antenna. We studied these in the Oleander hawk moth (Daphnis nerii) using a combination of techniques such as neural dye fills, confocal microscopy, scanning electron microscopy and X-ray tomography. Our study thus provides a detailed anatomical picture of the antennal mechanosensory-motor apparatus, which in turn provides key insights into its multifunctional role.


Developing neurons form transient nanotubes facilitating electrical coupling and calcium signaling with distant astrocytes.
Despite the well-documented cooperation between neurons and astrocytes little is known as to how these interactions are initiated. We show here by differential interference contrast microscopy that immature hippocampal neurons generated short protrusions towards astrocytes resulting in tunneling nanotube (TNT) formation with an average lifetime of 15 minutes. Fluorescence microscopy revealed that all TNTs between the two cell types contained microtubules but 35% of them were F-actin negative. Immunolabeling against connexin 43 showed that this gap junction marker localized at the contact site of TNTs with astrocytes. Using optical membrane-potential measurements combined with mechanical stimulation, we observed that ~35% of immature neurons were electrically coupled with distant astrocytes via TNTs up to 5 hours after co-culture but not after 24 hours. Connexin 43 was expressed by most neurons at 5 hours of co-culture but was not detected in neurons after 24 hours. We show that TNTs mediated the propagation of both depolarization and transient calcium signals from distant astrocytes to neurons. Our findings suggest that within a limited maturation period developing neurons establish electrical coupling and exchange of calcium signals with astrocytes via TNTs, which correlates with a high neuronal expression level of connexin 43. This novel cell-cell communication pathway between cells of the central nervous system provides new concepts in our understanding of neuronal migration and differentiation.


Association of health beliefs and colonoscopy use among survivors of colorectal cancer.
Clinical practice guidelines recommend ongoing testing (surveillance) for colorectal cancer survivors because they remain at risk for both local recurrences and second primary tumors. However, survivors often do not receive colorectal cancer surveillance. We used the Health Belief Model (HBM) to identify health beliefs that predict intentions to obtain routine colonoscopies among colorectal cancer survivors. We completed telephone interviews with 277 colorectal cancer survivors who were diagnosed 4 years earlier, between 2003 and 2005, in North Carolina. The interview measured health beliefs, past preventive behaviors, and intentions to have a routine colonoscopy in the next 5 years. In bivariate analyses, most HBM constructs were associated with intentions. In multivariable analyses, greater perceived likelihood of colorectal cancer (OR = 2.00, 95% CI = 1.16-3.44) was associated with greater intention to have a colonoscopy. Survivors who already had a colonoscopy since diagnosis also had greater intentions of having a colonoscopy in the future (OR = 9.47, 95% CI = 2.08-43.16). Perceived likelihood of colorectal cancer is an important target for further study and intervention to increase colorectal cancer surveillance among survivors. Other health beliefs were unrelated to intentions, suggesting that the health beliefs of colorectal cancer survivors and asymptomatic adults may differ due to the experience of cancer.


Skin-specific deletion of stearoyl-CoA desaturase-1 alters skin lipid composition and protects mice from high fat diet-induced obesity.
Stearoyl-CoA desaturase-1 (SCD1) catalyzes the synthesis of monounsaturated fatty acids and is an important regulator of whole body energy homeostasis. Severe cutaneous changes in mice globally deficient in SCD1 also indicate a role for SCD1 in maintaining skin lipids. We have generated mice with a skin-specific deletion of SCD1 (SKO) and report here that SKO mice display marked sebaceous gland hypoplasia and depletion of sebaceous lipids. In addition, SKO mice have significantly increased energy expenditure and are protected from high fat diet-induced obesity, thereby recapitulating the hypermetabolic phenotype of global SCD1 deficiency. Genes of fat oxidation, lipolysis, and thermogenesis, including uncoupling proteins and peroxisome proliferator-activated receptor-gamma co-activator-1alpha, are up-regulated in peripheral tissues of SKO mice. However, unlike mice globally deficient in SCD1, SKO mice have an intact hepatic lipogenic response to acute high carbohydrate feeding. Despite increased basal thermogenesis, SKO mice display severe cold intolerance because of rapid depletion of fuel substrates, including hepatic glycogen, to maintain core body temperature. These data collectively indicate that SKO mice have increased cold perception because of loss of insulating factors in the skin. This results in up-regulation of thermogenic processes for temperature maintenance at the expense of fuel economy, illustrating cross-talk between the skin and peripheral tissues in maintaining energy homeostasis.


A multisubunit factor, CstF, is required for polyadenylation of mammalian pre-mRNAs.
We have purified and characterized a factor required for accurate polyadenylation of mammalian pre-mRNAs in vitro. This factor, called cleavage-stimulation factor (CstF), is composed of three distinct polypeptide subunits of 77, 64, and 50 kD. Using monoclonal antibodies directed against the 64- and 50-kD subunits, we show that CstF is required for efficient cleavage of polyadenylation substrates. Furthermore, CstF present in unfractionated nuclear extracts interacts with pre-mRNAs containing the signal sequence AAUAAA, but not AAGAAA, in such a manner that the 64-kD subunit can be cross-linked to the RNA by UV light. This polypeptide is thus identical to the previously described 64-kD nuclear protein that binds to AAUAAA-containing RNAs. Finally, indirect immunofluorescence of fixed cells indicates that CstF is distributed diffusely throughout the nucleus in a granular pattern distinct from the "speckled" pattern displayed by factors involved in pre-mRNA splicing, but similar to that of heterogeneous nuclear ribonucleoproteins. A model is presented in which CstF binds specifically to nascent RNA polymerase II transcripts and, by interacting with other factors, results in a rapid initiation of 3'-end processing of pre-mRNAs.


Distribution of tissue progenitors within the shield region of the zebrafish gastrula.
The zebrafish has emerged as an important model system for the experimental analysis of vertebrate development because it is amenable to genetic analysis and because its optical clarity allows the movements and the differentiation of individual cells to be followed in vivo. In this paper, we have sought to characterize the spatial distribution of tissue progenitors within the outer cell layers of the embryonic shield region of the early gastrula. Single cells were labeled by iontophoretic injection of fluorescent dextrans. Subsequently, we documented their position with respect to the embryonic shield and their eventual fates. Our data show that progenitor cells of the neural, notochordal, somitic and endodermal lineages were all present within the embryonic shield region, and that these progenitors were arranged as intermingled populations. Moreover, close to the midline, there was evidence for significant biases in the distribution of neural and notochord progenitors between the layers, suggesting some degree of radial organization within the zebrafish embryonic shield region. The distributions of tissue progenitors in the zebrafish gastrula differ significantly from those in amphibians; this bears not only on interpretations of mutant phenotypes and in situ staining patterns, but also on our understanding of morphogenetic movements during gastrulation and of neural induction in the zebrafish.