Evo-Devo: evolutionary developmental mechanisms.
Evolutionary developmental biology (Evo-Devo) as a discipline is concerned, among other things, with discovering and understanding the role of changes in developmental mechanisms in the evolutionary origin of aspects of the phenotype. In a very real sense, Evo-Devo opens the black box between genotype and phenotype, or more properly, phenotypes as multiple life history stages arise in many organisms from a single genotype. Changes in the timing or positioning of an aspect of development in a descendant relative to an ancestor (heterochrony and heterotopy) were two evolutionary developmental mechanisms identified by Ernst Haeckel in the 1870s. Many more have since been identified, in large part because of our enhanced understanding of development and because new mechanisms emerge as development proceeds: the transfer from maternal to zygotic genomic control; cell-to-cell interactions; cell differentiation and cell migration; embryonic inductions; functional interactions at the tissue and organ levels; growth. Within these emergent processes, gene networks and gene cascades (genetic modules) link the genotype with morphogenetic units (cellular modules, namely germ layers, embryonic fields or cellular condensations), while epigenetic processes such as embryonic inductions, tissue interactions and functional integration, link morphogenetic units to the phenotype. Evolutionary developmental mechanisms also include interactions between individuals of the same species, individuals of different species, and species and their biotic and/or abiotic environment. Such interactions link ecological communities. Importantly, there is little to distinguish the causality that underlies these interactions from that which underlies inductive interactions within embryos.


Coffee consumption and risk of gastric cancer: an updated meta-analysis.
Coffee is one of the most widely consumed beverages worldwide, and many studies have investigated the association between coffee consumption and gastric cancer. However, the results are inconsistent. We conducted a systematic analysis of relevant population studies to derive a more precise estimation. Cochrane library, PubMed and Embase databases were searched to identify studies that met predetermined inclusion criterion through July 2014. All epidemiologic studies regarding coffee consumption and gastric cancer risk were selected, and relative risks (RRs) with 95% confidence intervals (CIs) were calculated. Twenty two studies (9 cohort and 13 case-control studies) involving 7,631 cases and 1,019,693 controls were included. The summary RR of gastric cancer was 0.94 (95% CI: 0.80-1.10) for the highest category of coffee consumption compared with the lowest category, and 0.93 (95% CI: 0.88-0.99) for coffee drinkers compared with nondrinkers. We stratified the population by coffee consumption. The pooled RR for the population with <1 cup/day, 1-2 cups/day and 3-4 cups/day coffee consumption compared with nondrinkers were 0.95 (95% CI: 0.84-1.08), 0.92 (95% CI: 0.82-1.03) and 0.88 (95% CI: 0.76-1.02), respectively, indicating that an increase in coffee consumption was associated with a decreased risk of gastric cancer. Furthermore, we stratified the studies by design, sex, population and time. A significant association between coffee intake and decreased gastric cancer risk was shown in case-control studies (RR=0.85, 95% CI: 0.77-0.95) and among the studies published over the last ten years (RR=0.88, 95% CI: 0.77-1.00). Our meta-analysis suggested that coffee consumption might be associated with a decreased risk of gastric cancer.


A facet-controlled Rh3Pb2S2 nanocage as an efficient and robust electrocatalyst toward the hydrogen evolution reaction.
Highly active and durable electrocatalysts for the hydrogen evolution reaction (HER) may play a pivotal role in commercial success of electrolytic water splitting technology. Among various material classes, binary metal sulphides show a great promise as HER catalysts because of their tunable energy levels conducive to a high catalytic activity and high robustness under harsh operating conditions. On the other hand, facet-controlled nanoparticles with controlled surface energies have gained great recent popularity as active and selective catalysts. However, binary metal sulphide nanoparticles with well-defined facets and high surface areas are very rare. Herein we report the synthesis of a facet-controlled hollow Rh3Pb2S2 nanocage as a new catalytic material and its excellent activity (overpotential: 87.3 mV at 10 mA cm-2) and robustness toward HER under harsh acidic conditions.


Falsely elevated thyroid hormone levels caused by anti-ruthenium interference in the Elecsys assay resembling the syndrome of inappropriate secretion of thyrotropin.
The syndrome of inappropriate secretion of thyrotropin (SITSH) is defined as the inappropriate non-suppression of serum TSH in the presence of elevated free thyroid hormone; TSH-secreting pituitary adenomas and the syndrome of resistance to thyroid hormone are the main etiologies of SITSH. In addition, erroneous thyroid function testing may result in the diagnosis of this syndrome. A 63-year-old woman was referred because of suspected SITSH. Laboratory tests showed a normal TSH (0.52 μIU/L; normal range: 0.5-5.0) measured by sandwich Elecsys, and elevated FT4 (3.8 ng/dL; normal range: 0.9-1.6) and FT3 (7.6 pg/mL; normal range: 2.3-4.0), determined by competitive Elecsys. To exclude possible assay interference, aliquots of the original samples were retested using a different method (ADVIA Centaur), which showed normal FT4 and FT3 levels. Eight hormone levels, other than thyroid function tests measured by competitive or sandwich Elecsys, were higher or lower than levels determined by an alternative analysis. Subsequent examinations, including gel filtration chromatography, suggested interference by substances against ruthenium, which reduced the excitation of ruthenium, and resulted in erroneous results. The frequency of similar cases, where the FT4 was higher than 3.2 ng/dL, in spite of a non-suppressed TSH, was examined; none of 10 such subjects appeared to have method-specific interference. Here, a patient with anti-ruthenium interference, whose initial thyroid function tests were consistent with SITSH, is presented. This type of interference should be considered when thyroid function is measured using the Elecsys technique, although the frequency of such findings is likely very low.


Emergency department triage: a program assessment using the tools of continuous quality improvement.
An assessment was undertaken in the emergency department of a busy tertiary care center to illustrate the role of continuous quality improvement in the evaluation of an emergency triage program that utilizes the emergency medical attendant to provide triage. An evaluation team interviewed triage staff, charge nurses, internal customers, risk management, and the patient representative. A detailed review of staff job descriptions, organization charts, orientation manual, and physical facilities was conducted. A chart audit was completed on 100 triage notes. Direct observation was undertaken on nine occasions. An evaluation of the data gathered was performed using the tools of continuous quality improvement, and resulted in specific recommendations being made to improve the process of care. It was concluded that emergency medical attendants function very well in an emergency medicine triage system and the tools of continuous quality improvement can be applied to a clinical service to improve the quality of care.


eNOS gene polymorphisms in paraffin-embedded tissues of prostate cancer patients.
The purpose of the present study was to investigate whether endothelial nitric oxide synthase (eNOS) gene polymorphisms play a role in prostate cancer (PCa). We examined three eNOS gene polymorphisms (T-786C promoter region, G894T, and Intron 4 VNTR 4a/b) at extracted DNAs from 50 formalin-fixed paraffin-embedded tissues of PCa patients. For the controls, blood samples obtained from 50 healthy men were studied. Genotyping of molecular variants was performed by PCR-RFLP technique. We found that the TC genotype of the T-786C polymorphism was associated with PCa risk (OR: 3.325, CI: 1.350-8.188, P = 0.008). The eNOS G894T polymorphism was also associated with PCa. The frequency of the 894T allele was significantly higher in PCa patients. No association was identified between intron 4 VNTR polymorphism and PCa. We found significant differences in genotypic and allelic frequencies between PCa patients and controls for eNOS T-786C and G894T polymorphisms. The presence of the T-786C genotype and 894T allele in carriers increased the risk of PCa. No association was found between intron 4 VNTR polymorphism and PCa patients.


Habitat selection in a rocky landscape: experimentally decoupling the influence of retreat site attributes from that of landscape features.
Organisms selecting retreat sites may evaluate not only the quality of the specific shelter, but also the proximity of that site to resources in the surrounding area. Distinguishing between habitat selection at these two spatial scales is complicated by co-variation among microhabitat factors (i.e., the attributes of individual retreat sites often correlate with their proximity to landscape features). Disentangling this co-variation may facilitate the restoration or conservation of threatened systems. To experimentally examine the role of landscape attributes in determining retreat-site quality for saxicolous ectotherms, we deployed 198 identical artificial rocks in open (sun-exposed) sites on sandstone outcrops in southeastern Australia, and recorded faunal usage of those retreat sites over the next 29 months. Several landscape-scale attributes were associated with occupancy of experimental rocks, but different features were important for different species. For example, endangered broad-headed snakes (Hoplocephalus bungaroides) preferred retreat sites close to cliff edges, flat rock spiders (Hemicloea major) preferred small outcrops, and velvet geckos (Oedura lesueurii) preferred rocks close to the cliff edge with higher-than-average sun exposure. Standardized retreat sites can provide robust experimental data on the effects of landscape-scale attributes on retreat site selection, revealing interspecific divergences among sympatric taxa that use similar habitats.


Induction of synthesis of components of the hydrogen peroxide-generating oxidase during activation of the human monocytic cell line U937 by interferon-gamma.
Increased amounts of cytochrome b-245 and a 45 kDa polypeptide component of the hydrogen peroxide-generating oxidase were detected in the human monocytic cell line U937 after incubation in interferon-gamma. The time course of increase in cytochrome b-245 paralleled the induction of oxidase activity. Induction of the 45 kDa component had a different time course, having a greater lag before increased amounts were seen.


An androgen receptor mutation in the MDA-MB-453 cell line model of molecular apocrine breast cancer compromises receptor activity.
Recent evidence indicates that the estrogen receptor-α-negative, androgen receptor (AR)-positive molecular apocrine subtype of breast cancer is driven by AR signaling. The MDA-MB-453 cell line is the prototypical model of this breast cancer subtype; its proliferation is stimulated by androgens such as 5α-dihydrotestosterone (DHT) but inhibited by the progestin medroxyprogesterone acetate (MPA) via AR-mediated mechanisms. We report here that the AR gene in MDA-MB-453 cells contains a G-T transversion in exon 7, resulting in a receptor variant with a glutamine to histidine substitution at amino acid 865 (Q865H) in the ligand binding domain. Compared with wild-type AR, the Q865H variant exhibited reduced sensitivity to DHT and MPA in transactivation assays in MDA-MB-453 and PC-3 cells but did not respond to non-androgenic ligands or receptor antagonists. Ligand binding, molecular modeling, mammalian two-hybrid and immunoblot assays revealed effects of the Q865H mutation on ligand dissociation, AR intramolecular interactions, and receptor stability. Microarray expression profiling demonstrated that DHT and MPA regulate distinct transcriptional programs in MDA-MB-453 cells. Gene Set Enrichment Analysis revealed that DHT- but not MPA-regulated genes were associated with estrogen-responsive transcriptomes from MCF-7 cells and the Wnt signaling pathway. These findings suggest that the divergent proliferative responses of MDA-MB-453 cells to DHT and MPA result from the different genetic programs elicited by these two ligands through the AR-Q865H variant. This work highlights the necessity to characterize additional models of molecular apocrine breast cancer to determine the precise role of AR signaling in this breast cancer subtype.


In-Person and Telehealth Ambulatory Contacts and Costs in a Large US Insured Cohort Before and During the COVID-19 Pandemic.
This study assesses the role of telehealth in the delivery of care at the start of the COVID-19 pandemic. To document patterns and costs of ambulatory care in the US before and during the initial stage of the pandemic and to assess how patient, practitioner, community, and COVID-19-related factors are associated with telehealth adoption. This is a cohort study of working-age persons continuously enrolled in private health plans from March 2019 through June 2020. The comparison periods were March to June in 2019 and 2020. Claims data files were provided by Blue Health Intelligence, an independent licensee of the Blue Cross and Blue Shield Association. Data analysis was performed from June to October 2020. Ambulatory encounters (in-person and telehealth) and allowed charges, stratified by characteristics derived from enrollment files, practitioner claims, and community characteristics linked to the enrollee's zip code. A total of 36 568 010 individuals (mean [SD] age, 35.71 [18.77] years; 18 466 557 female individuals [50.5%]) were included in the analysis. In-person contacts decreased by 37% (from 1.63 to 1.02 contacts per enrollee) from 2019 to 2020. During 2020, telehealth visits (0.32 visit per person) accounted for 23.6% of all interactions compared with 0.3% of contacts in 2019. When these virtual contacts were added, the overall COVID-19 era patient and practitioner visit rate was 18% lower than that in 2019 (1.34 vs 1.64 visits per person). Behavioral health encounters were far more likely than medical contacts to take place virtually (46.1% vs 22.1%). COVID-19 prevalence in an area was associated with higher use of telehealth; patients from areas within the top quintile of COVID-19 prevalence during the week of their encounter were 1.34 times more likely to have a telehealth visit compared with those in the lowest quintile (the reference category). Persons living in areas with limited social resources were less likely to use telehealth (most vs least socially advantaged neighborhoods, 27.4% vs 19.9% usage rates). Per enrollee medical care costs decreased by 15% between 2019 and 2020 (from $358.32 to $306.04 per person per month). During 2020, those with 1 or more COVID-19-related service (1 470 721 members) had more than 3 times the medical costs ($1701 vs $544 per member per month) than those without COVID-19-related services. Persons with 1 or more telehealth visits in 2020 had considerably higher costs than persons having only in-person ambulatory contacts ($2214.10 vs $1337.78 for the COVID-19-related subgroup and $735.87 vs $456.41 for the non-COVID-19 subgroup). This study of a large cohort of patients enrolled in US health plans documented patterns of care at the onset of COVID-19. The findings are relevant to policy makers, payers, and practitioners as they manage the use of telehealth during the pandemic and afterward.


Influence of the interaction of light- and self-polymerization on subsurface hardening of a dual-cured core build-up resin composite.
To investigate the influence of time delay and duration of photo-activation on subsurface microhardness of a dual-cured resin composite. A commercially available dual-cured core build-up resin composite (Rebilda DC) was filled in cavities (diameter: 4.0 mm; height: 6.0 mm) of polystyrene molds and light-cured for 20 or 60 s either immediately after the filling procedure (time delay 0 s) or after a time delay of 30, 90, 180 or 300 s. Non-irradiated self-cured specimens served as a control group (n = 15). Specimens were stored in complete darkness and at 100% relative humidity at 37°C for 2 weeks and cross-sectioned. Knoop Hardness Numbers (KHNs) were measured six times per depth and averaged at distances of 0.25, 0.50, 1.00, 2.00, 3.50 and 5.50 mm from the light-exposed surface. Data were statistically analyzed using one- and two-way ANOVA followed by Scheffé's post-hoc test at a level of significance of 0.05. Mean hardness values in all experimental groups ranged between 54.3 ± 2.1 and 58.1 ± 2.3 KHN. Light-curing did not significantly increase composite KHN at any depth measured. Delaying light exposure had no influence on KHN, irrespective of depth. A longer light-exposure time (60 versus 20 s) resulted in significantly higher KHN only at depths of 3.50 and 5.50 mm. Photo-activation of the tested dual-cured resin composite provided no clinically relevant benefit compared to self-curing regarding the degree of hardening.


Insulin-degrading enzyme rapidly removes the beta-amyloid precursor protein intracellular domain (AICD).
The intramembranous gamma-secretase cleavage of the beta-amyloid precursor protein (APP) is dependent on biologically active presenilins (PS). Notch also undergoes a similar PS-dependent gamma-secretase-like cleavage, resulting in the liberation of the Notch intracellular domain (NICD), which is critically required for developmental signal transduction. gamma-Secretase processing of APP results in the production of a similar fragment called AICD (APP intracellular domain), which may function in nuclear signaling as well. AICD, like NICD, is rapidly removed. By using a battery of protease inhibitors we demonstrate that AICD, in contrast to NICD, is degraded by a cytoplasmic metalloprotease. In vitro degradation of AICD can be reconstituted with cytoplasmic fractions obtained from neuronal and non-neuronal cells. Taking into account the inhibition profile and the cytoplasmic localization, we identified three candidate enzymes (neurolysin, thimet oligopeptidase, and insulin-degrading enzyme (IDE), also known as insulysin), which all are involved in the degradation of bioactive peptides in the brain. When insulin, a well characterized substrate of IDE, was added to the in vitro degradation assay, removal of AICD was efficiently blocked. Moreover, overexpression of IDE resulted in enhanced degradation of AICD, whereas overexpression of the inactive IDE E111Q mutant did not affect AICD degradation. Finally, immunodepletion of IDE significantly reduced the AICD degrading activity. Therefore our data demonstrate that IDE, which is one of the proteases implicated in the removal of extracellular Abeta, also removes the cytoplasmic product of gamma-secretase cleaved APP.


Coronary computed tomographic angiography and risk of all-cause mortality and nonfatal myocardial infarction in subjects without chest pain syndrome from the CONFIRM Registry (coronary CT angiography evaluation for clinical outcomes: an international multicenter registry).
The predictive value of coronary computed tomographic angiography (cCTA) in subjects without chest pain syndrome (CPS) has not been established. We investigated the prognostic value of coronary artery disease detection by cCTA and determined the incremental risk stratification benefit of cCTA findings compared with clinical risk factor scoring and coronary artery calcium scoring (CACS) for individuals without CPS. An open-label, 12-center, 6-country observational registry of 27 125 consecutive patients undergoing cCTA and CACS was queried, and 7590 individuals without CPS or history of coronary artery disease met the inclusion criteria. All-cause mortality and the composite of all-cause mortality and nonfatal myocardial infarction were measured. During a median follow-up of 24 months (interquartile range, 18-35 months), all-cause mortality occurred in 136 individuals. After risk adjustment, compared with individuals without evidence of coronary artery disease by cCTA, individuals with obstructive 2- and 3-vessel disease or left main coronary artery disease experienced higher rates of death and composite outcome (P<0.05 for both). Both CACS and cCTA significantly improved the performance of standard risk factor prediction models for all-cause mortality and the composite outcome (likelihood ratio P<0.05 for all), but the incremental discriminatory value associated with their inclusion was more pronounced for the composite outcome and for CACS (C statistic for model with risk factors only was 0.71; for risk factors plus CACS, 0.75; for risk factors plus CACS plus cCTA, 0.77). The net reclassification improvement resulting from the addition of cCTA to a model based on standard risk factors and CACS was negligible. Although the prognosis for individuals without CPS is stratified by cCTA, the additional risk-predictive advantage by cCTA is not clinically meaningful compared with a risk model based on CACS. Therefore, at present, the application of cCTA for risk assessment of individuals without CPS should not be justified.


Girls on a high-calcium diet gain weight at the same rate as girls on a normal diet: a pilot study.
To determine whether pubertal girls assigned to calcium-rich diets or their usual calcium intakes differ significantly in weight gain. This is a pilot study conducted on data from an experimental research project. The sample included 59 girls, 9 years of age, from Omaha metropolitan communities. Participants were randomly assigned to a calcium-rich diet supplying at least 1,500 mg of calcium per day or their usual diet. The outcome measure was change in weight during 2 years of study. Data were analyzed using descriptive statistics, Mann-Whitney U, t tests of paired and independent samples, and analysis of covariance. The girls in the calcium-rich diet group had a mean (+/-standard deviation) calcium intake of 1,656+/-191 mg/day, whereas the girls on their usual diets averaged 961+/-268 mg/day. Although the participants in the treatment group consumed nearly twice as much dietary calcium--primarily from dairy foods--they did not have greater increases in body weight, body mass index, or fat or lean mass than the control group. These findings held when the data were grouped by tertile of calcium intake. Compared with girls on their usual diets, the girls who consumed the calcium-rich diet also significantly increased their intake of essential nutrients. We conclude that calcium-rich diets do not cause excessive weight gain in pubertal girls but do contribute positively to overall nutrition. These findings provide support for programs to promote calcium-rich diets, which are critical for attaining peak bone mass.


Low-dose lipopolysaccharide (LPS) pretreatment of mouse macrophages modulates LPS-dependent interleukin-6 production in vitro.
Lipopolysaccharide (LPS) can induce mouse macrophages to produce a number of cytokines and other inflammatory mediators. Our laboratory previously reported that LPS-dependent macrophage-derived tumor necrosis factor alpha (TNF-alpha) production could be significantly potentiated by pretreatment with LPS at substimulatory LPS priming doses. The observed potentiation was shown to be coincident with a down-regulation of LPS-dependent nitric oxide (NO) production (X. Zhang and D. C. Morrison, J. Exp. Med. 177: 511-516, 1993). In order to determine whether these LPS reprogramming effects in mouse macrophages were selective for these two macrophage-derived mediators, we have examined the effects of LPS pretreatment on LPS-dependent interleukin 6 (IL-6) production. Thioglycolate-elicited mouse peritoneal macrophages were pretreated with various subthreshold stimulatory concentrations of LPS for 6 h, washed three times, and then stimulated with an effective stimulatory concentration of smooth LPS for 18 h. In confirmation of earlier studies, pretreatment of mouse macrophages with substimulatory doses of LPS inhibited the subsequent LPS-dependent NO production. This down-regulation was accompanied by a coordinate up-regulation of LPS-dependent IL-6 production, similar to what was shown earlier for TNF-alpha production. These priming effects with the substimulatory dose of smooth LPS are shown to be independent of doses of LPS used for subsequent activation and are not restricted to specific LPS stimulation. Moreover, the enhancement of the IL-6 response by LPS pretreatment is still observed in the presence of neutralizing antibody to TNF-alpha. These findings, therefore, provide further support for the conclusion that LPS-dependent macrophage reprogramming is likely to involve common regulatory pathways that control the secretion of both IL-6 and TNF-alpha.


Buprenorphine prescribing practice trends and attitudes among New York providers.
Buprenorphine office-based opioid maintenance is an increasingly common form of treatment for opioid use disorders. However, total prescribing has not kept pace with the current opioid and overdose epidemic and access remains scarce among the underserved. This study sought to assess current provider attitudes and clinical practices among a targeted sample of primarily New York City public sector buprenorphine prescribers. A cross-sectional online survey purposefully sampled buprenorphine prescribers in NYC with a focus on those serving Medicaid and uninsured patient populations. Expert review of local provider networks, snowball referrals, and in-person networking generated an email list, which received a survey link. A brief 25-question instrument queried provider and practice demographics, prescribing practices including induction approaches and attitudes regarding common hot topics (e.g., buprenorphine diversion, prescriber patient limits, insurance issues, ancillary treatments). Of 132 email invitations, N=72 respondents completed (n=64) or partially completed (n=8) the survey between January and April 2016. Most (79%) were Medicaid providers in non-psychiatric specialties (72%), working in a hospital-based or community general practice (51%), and board-certified in addiction medicine or psychiatry (58%). Practice sizes were generally 100 patients or fewer (71%); many providers (64%) individually prescribed buprenorphine <25% of total practice time to a median 23 patients (mean 31, range 0-102). Unobserved (home) induction for new patients was a common practice: 49% predominantly prescribed unobserved induction; 16% mixed unobserved and observed inductions. Adjunctive psychosocial counseling was routinely recommended (46%) or considered on a case-by-case basis (17%) versus mandated (37%). Medication prior authorization requirements were the highest rated barriers to practice, followed by inadequate clinic space, limited clinic time and/or support staff, and inadequate psychiatric services for dual diagnoses. Buprenorphine diversion was not rated as an important practice barrier. In conclusion, this targeted survey of buprenorphine prescribers in NYC treating primarily underserved populations showed a consistent pattern of part-time prescribing to modest volumes of patients, routine use of unobserved buprenorphine induction, and primarily elective referrals to psychosocial counseling. Barriers to prescribing included prior authorization requirements, lack of clinical resources (space, staff) and psychiatric services. Federal and local efforts to reduce such barriers may improve buprenorphine access among the underserved.


Protocol for Initiating and Monitoring Bumble Bee Microcolonies with Bombus impatiens (Hymenoptera: Apidae).
Populations of some bumble bee species are in decline, prompting the need to better understand bumble bee biology and for assessing the effects of environmental stressors on these important pollinators. Microcolonies have been successfully used for investigating a range of endpoints, including behavior, gut microbiome, nutrition, development, pathogens, and the effects of pesticide exposure on bumble bee health. Here, we present a step-by-step protocol for initiating, maintaining, and monitoring microcolonies with Bombus impatiens . This protocol has been successfully used in two pesticide exposure-effects studies and can be easily expanded to investigate other aspects of bumble bee biology.


Dutch home-based pre-reading intervention with children at familial risk of dyslexia.
Children (5 and 6 years old, n = 30) at familial risk of dyslexia received a home-based intervention that focused on phoneme awareness and letter knowledge in the year prior to formal reading instruction. The children were compared to a no-training at-risk control group (n = 27), which was selected a year earlier. After training, we found a small effect on a composite score of phoneme awareness (d = 0.29) and a large effect on receptive letter knowledge (d = 0.88). In first grade, however, this did not result in beneficial effects for the experimental group in word reading and spelling. Results are compared to three former intervention studies in The Netherlands and comparable studies from Denmark and Australia.


Interaction between the N-terminus of human topoisomerase I and SV40 large T antigen.
We have attempted to identify human topoisomerase I-binding proteins in order to gain information regarding the cellular roles of this protein and the cytotoxic mechanisms of the anticancer drug camptothecin, which specifically targets topoisomerase I. In the course of this work we identified an interaction between the N-terminus of human topoisomerase I and the SV40 T antigen that is detectable in vitro using both affinity chromatography and co-immunoprecipitation. Additional results indicate that this interaction does not require intermediary DNA or stoichiometric quantities of other proteins. Furthermore, the interaction is detectable in vivo using a yeast two-hybrid assay. Two binding sites for T antigen are apparent on the topoisomerase I protein: one consisting of amino acids 1-139, the other present in the 383-765 region of the protein. Interestingly, nucleolin, which binds the 166-210 region of topoisomerase I, is able to bind an N-terminal fragment of topoisomerase I concurrently with T antigen. Taken together with our prior identification of nucleolin as a topoisomerase I-binding protein, the current results suggest that helicase-binding is a major role of the N-terminus of human topoisomerase I and that the resultant helicase-topoisomerase complex may function as a eukaryotic gyrase.


A mathematical model for a new mechanism of phenotypic variation in malaria.
The Py235 merozoite rhoptry protein of the rodent malaria agent Plasmodium (yoelii) yoeli is encoded by the Py235 multigene family whose members are transcribed during the parasite's asexual life-cycle in a fashion where single schizonts subsequently give rise to sets of merozoites containing distinct Py235 transcripts. Homologues of Py235 are found in other malaria species, and antibodies to both Py235 and P. falciparum homologues inhibit merozoite invasion, suggesting a unique survival strategy involving immune evasion and host adaptation. Using a mathematical approach to model this free-living stage of Plasmodium in interaction with specific antibodies and a heterogeneous red blood cell population, we investigate if, and under what conditions, this mechanism of clonal phenotypic variation can play a role in immune evasion and adaptation to a dynamic erythropoietic environment.


[Ideal number of children as a risk factor for tubal ligation].
The purpose of this paper was to evaluate the association between ideal number of children (INC) and female sterilization. A nested case-control study was performed through a secondary analysis of data from a cohort study on the reproductive health of women in Campinas, São Paulo, Brazil. A total of 3,878 women were included, 1,012 being sterilized (cases). The relationship between INC and number of live births (LB) was divided in two categories (INC > LB and INC < or = LB). The relative risks of performing tubal ligation were calculated (odds ratio) with their respective 95% confidence intervals for the relation INC/LB and all control variables. All predictor variables were included in a logistic regression model in order to identify the factors independently associated with female sterilization. The results showed that the risks of tubal ligation were higher among women with INC < or = LB, higher age, with partners, higher family income, more than two previous pregnancies, more deliveries, fewer abortions, and without paid work.


Characterization of carbapenem nonsusceptible Pseudomonas aeruginosa in Denmark: a nationwide, prospective study.
From January 1st 2011 through June 30th 2011, 116 nonreplicate, noncystic fibrosis-related Pseudomonas aeruginosa isolates with reduced carbapenem susceptibility were collected from 12 out of 13 Danish departments of clinical microbiology. The presence of acquired β-lactamases was assessed with combination tablet-diffusion methodology and polymerase chain reaction. In addition, antimicrobial susceptibility testing, an efflux pump inhibitor assay, and pulsed-field gel electrophoresis (PFGE) were performed. Isolates producing acquired β-lactamases were further investigated by serotyping and multi locus sequence typing. Eight isolates produced the metallo-β-lactamase (MBL) VIM-2, and one isolate produced OXA-10 and VEB-1-like extended-spectrum beta-lactamase (ESBL). Phenotypic indications of derepressed AmpC and efflux pump were seen in 56 and 43 isolates, respectively. Overall, the results indicate that mutational factors related to permeability--often combined with derepressed, chromosomal AmpC--is the main factor behind carbapenem nonsusceptibility in Danish P. aeruginosa isolates. The ESBL producer and all the VIM producers belonged to international clones. PFGE revealed that most of the isolates were unrelated, but clonal spread was seen; the 116 isolates distributed in 97 PFGE types, with the largest cluster consisting of 4 isolates (including three isolates from the same hospital with 100% similarity). Thirty-two isolates were pair-wise related, while the remaining isolates were clonally unrelated, as were all nine ESBL/MBL producers.


Novel bead-based platform for direct detection of unlabelled nucleic acids through Single Nucleobase Labelling.
Over the last decade, circulating microRNAs have received attention as diagnostic and prognostic biomarkers. In particular, microRNA122 has been demonstrated to be an early and more sensitive indicator of drug-induced liver injury than the widely used biomarkers such as alanine aminotransferase and aspartate aminotransferase. Recently, microRNA122 has been used in vitro to assess the cellular toxicity of new drugs and as a biomarker for the development of a rapid test for drug overdose/liver damage. In this proof-of-concept study, we report a PCR-free and label-free detection method that has a limit of detection (3 standard deviations) of 15 fmoles of microRNA122, by integrating a dynamic chemical approach for "Single Nucleobase Labelling" with a bead-based platform (Luminex®) thereby, in principle, demonstrating the exciting prospect of rapid and accurate profiling of any microRNAs related to diseases and toxicology.


Chemical aspects of clustered DNA damage induction by ionising radiation.
Ionising radiation induces a variety of chemical modifications to DNA, ranging from simple, isolated lesions to clustered DNA damage, in which two or more lesions are formed within a few tens of base pairs by a single radiation track. Multiple lesions, e.g. tandem lesions and amplification of damage, may also be induced in DNA by reaction with a single hydroxyl radical. It has been proposed from biophysical modelling that clustered DNA damage is less repairable and therefore contributes to the biological severity of ionising radiation. In this review, some evidence is presented which indicates that non-double strand break (non-DSB) clustered DNA damage is induced in significant yield, relative to that of DSBs, in mammalian cells. Enzymatic processing of clustered DNA damage in synthetic oligonucleotides has been shown to be compromised, depending on the nature of the lesions present. The role of clustered DNA damage in the early stages of the development of radiation-induced carcinogenesis remains to be addressed.


Alcohol use patterns among Brazilian adults.
To describe patterns of alcohol consumption in the adult Brazilian population. Multicluster random sample of 2,346 subjects 18 years of age or older, selected nationwide. Interviews were conducted in respondents' homes between November 2005 and April 2006. About 48% of the sample had not drunk alcoholic beverages in the past year, with variations by gender, age, marital status, education, income and region of the country. Among drinkers, 29% reported drinking 5 or more drinks per occasion (men, 38%). Sociodemographic variables are associated with the frequency and amount of drinking, alcohol problems, and alcohol abuse and dependence. Among the whole sample (including drinkers and non drinkers), 28% reported binge drinking, 25% reported at least one kind of alcohol related problem, 3% were alcohol abusers and another 9% were alcohol dependent. Abstinence is high in the Brazilian population. However, elevated proportions of those who drink consume alcohol in a high risk pattern (binge-drinking), report a high level of alcohol problems, alcohol abuse and dependence. National public policies must consider these factors, as well as the regional Brazilian differences.


Social Media Mining for an Analysis of Nutrition and Dietary Health in Taiwan.
Dining is an essential part of human life. In order to pursue a healthier self, more and more people enjoy homemade cuisines. Consequently, the amount of recipe websites has increased significantly. These online recipes represent different cultures and cooking methods from various regions, and provide important indications on nutritional content. In recent years, the development of data science made data mining a popular research area. However, only a few researches in Taiwan have applied data mining in the studies of recipes and nutrients. Therefore, this work aims at utilizing machine learning models to discover health-related insights from recipes on social media. First, we collected over 15,000 Chinese recipes from the largest recipe website in Taiwan to build a recipe database. We then extracted information from this dataset through natural language processing methodologies so as to better understand the characteristics of various cuisines and ingredients. Thus, we can establish a classification model for the automatic categorization of recipes. We further performed cluster analysis for grouping nutrients to recognize the nutritional differences for each cluster and each cuisine type. The results showed that using the support vector machine (SVM) model can successfully classify recipes with an average F-score of 82%. We also analyzed the nutritional value of different cuisine categories and the possible health effects they may bring to the consumers. Our methods and findings can assist future work on extracting essential nutritional information from recipes and promoting healthier diets.


Bacteria isolated from the different developmental stages and larval organs of the obligate parasitic fly, Wohlfahrtia magnifica (Diptera: Sarcophagidae).
Wohlfahrtia magnifica (Diptera: Sarcophagidae) is the major myiasis-causing fly species in the whole of Eurasia for most important domestic animals. The aim of the present work was to obtain data on the culturable bacteria isolated under aerobic conditions from this fly: bacteria were isolated from all developmental stages (larvae, pupa, and imago) of Wohlfahrtia magnifica, and the third-stage larval organs were also sampled. To determine the possible antagonistic effects between the dominant bacterial groups, an antibiosis assay was carried out. Plating and isolation of bacteria was performed by classical microbiological methods. Characterization of the isolated strains was carried out via a polyphasic approach; classical phenotypic tests, chemotaxonomical examinations, and 16S rDNA sequence analyses were also applied. In the case of maggot macerate samples, members of the family Enterobacteriaceae were characteristic. Members of a new genus (Schineria) belonging to the gamma subdivision of proteobacteria were also isolated. According to our data, the shifts in the Schineria and Proteus populations within the larvae are strongly influenced by their interactions with each other and among the members of the family Enterobacteriaceae. The pupa and imago samples contained several other Gram-negative bacteria (Stenotrophomonas, Brevundimonas, etc.). Among Gram-positive bacteria, in all maggot macerate samples, members of the genus Bacillus and the Arthrobacter-Micrococcus group of actinobacteria were dominant (neither of them was a producer or sensitive to the compounds of other microorganisms), and bacteria related to the genus Corynebacterium were also found. From the larvae Aureobacterium liquefaciens and Enterococcus faecalis were isolated, and from the pupae Dietzia maris and Enterococcus faecalis. In the samples of third-stage larval organs, the dominant groups were the same as in the third-stage larval macerate sample; however, several additional genera/species were observed (Rhodococcus fascians, Streptomyces sp., Rathayibacter sp., Bacillus thuringiensis/cereus).


Scientific foundations for an IUCN Red List of ecosystems.
An understanding of risks to biodiversity is needed for planning action to slow current rates of decline and secure ecosystem services for future human use. Although the IUCN Red List criteria provide an effective assessment protocol for species, a standard global assessment of risks to higher levels of biodiversity is currently limited. In 2008, IUCN initiated development of risk assessment criteria to support a global Red List of ecosystems. We present a new conceptual model for ecosystem risk assessment founded on a synthesis of relevant ecological theories. To support the model, we review key elements of ecosystem definition and introduce the concept of ecosystem collapse, an analogue of species extinction. The model identifies four distributional and functional symptoms of ecosystem risk as a basis for assessment criteria: A) rates of decline in ecosystem distribution; B) restricted distributions with continuing declines or threats; C) rates of environmental (abiotic) degradation; and D) rates of disruption to biotic processes. A fifth criterion, E) quantitative estimates of the risk of ecosystem collapse, enables integrated assessment of multiple processes and provides a conceptual anchor for the other criteria. We present the theoretical rationale for the construction and interpretation of each criterion. The assessment protocol and threat categories mirror those of the IUCN Red List of species. A trial of the protocol on terrestrial, subterranean, freshwater and marine ecosystems from around the world shows that its concepts are workable and its outcomes are robust, that required data are available, and that results are consistent with assessments carried out by local experts and authorities. The new protocol provides a consistent, practical and theoretically grounded framework for establishing a systematic Red List of the world's ecosystems. This will complement the Red List of species and strengthen global capacity to report on and monitor the status of biodiversity.


Human immunodeficiency virus type 1 gag-specific mucosal immunity after oral immunization with papillomavirus pseudoviruses encoding gag.
Mucosal surfaces are the primary portals for human immunodeficiency virus (HIV) transmission. Because systemic immunization, in general, does not induce effective mucosal immune responses, a mucosal HIV vaccine is urgently needed. For this study, we developed papillomavirus pseudoviruses that express HIV-1 Gag. The pseudoviruses are synthetic, nonreplicating viruses, yet they can produce antigens for a long time in the immune system. Here we show that oral immunization of mice by the use of papillomavirus pseudoviruses encoding Gag generated mucosal and systemic Gag-specific cytotoxic T lymphocytes that effectively lysed Gag-expressing target cells. Furthermore, the pseudoviruses generated Gag-specific gamma interferon-producing T cells and serum immunoglobulin G (IgG) and mucosal IgA. In contrast, oral immunization with plasmid DNA encoding HIV-1 Gag did not induce specific immune responses. Importantly, oral immunization with the pseudoviruses induced Gag-specific memory cytotoxic T lymphocytes and protected mice against a rectal mucosal challenge with a recombinant vaccinia virus expressing HIV-1 Gag. Thus, papillomavirus pseudoviruses encoding Gag are a promising mucosal vaccine against AIDS.


Combined analgesics in (headache) pain therapy: shotgun approach or precise multi-target therapeutics?
Pain in general and headache in particular are characterized by a change in activity in brain areas involved in pain processing. The therapeutic challenge is to identify drugs with molecular targets that restore the healthy state, resulting in meaningful pain relief or even freedom from pain. Different aspects of pain perception, i.e. sensory and affective components, also explain why there is not just one single target structure for therapeutic approaches to pain. A network of brain areas ("pain matrix") are involved in pain perception and pain control. This diversification of the pain system explains why a wide range of molecularly different substances can be used in the treatment of different pain states and why in recent years more and more studies have described a superior efficacy of a precise multi-target combination therapy compared to therapy with monotherapeutics. In this article, we discuss the available literature on the effects of several fixed-dose combinations in the treatment of headaches and discuss the evidence in support of the role of combination therapy in the pharmacotherapy of pain, particularly of headaches. The scientific rationale behind multi-target combinations is the therapeutic benefit that could not be achieved by the individual constituents and that the single substances of the combinations act together additively or even multiplicatively and cooperate to achieve a completeness of the desired therapeutic effect.As an example the fixed-dose combination of acetylsalicylic acid (ASA), paracetamol (acetaminophen) and caffeine is reviewed in detail. The major advantage of using such a fixed combination is that the active ingredients act on different but distinct molecular targets and thus are able to act on more signalling cascades involved in pain than most single analgesics without adding more side effects to the therapy. Multitarget therapeutics like combined analgesics broaden the array of therapeutic options, enable the completeness of the therapeutic effect, and allow doctors (and, in self-medication with OTC medications, the patients themselves) to customize treatment to the patient's specific needs. There is substantial clinical evidence that such a multi-component therapy is more effective than mono-component therapies.


Recommendations for Identifying Sleep Problems and Treatment Resources for Children with Autism Spectrum Disorder.
Many young children experience sleep problems that may influence their daytime functioning. These sleep problems are especially prevalent in young children with autism spectrum disorder (ASD). Therefore, general recommendations for identifying and treating sleep problems in children with ASD are needed for behavior analysts to identify potential sleep problems and make empirically informed decisions regarding treatment options. The current paper seeks to provide behavior analysts, who work with children with ASD, with informative research on pediatric sleep problems, sleep measures, and options for behavioral sleep treatment.


Metformin stimulates IGFBP-2 gene expression through PPARalpha in diabetic states.
The anti-diabetic drug, metformin, exerts its action through AMP-activated protein kinase (AMPK), and Sirtuin (Sirt1) signaling. Insulin-like growth factor (IGF)-binding protein 2 (IGFBP-2) prevents IGF-1 binding to its receptors, thereby contributing to modulate insulin sensitivity. In this study, we demonstrate that metformin upregulates Igfbp-2 expression through the AMPK-Sirt1-PPARα cascade pathway. In the liver of high fat diet, ob/ob, and db/db mice, Igfbp-2 expression was significantly decreased compared to the expression levels in the wild-type mice (p < 0.05). Upregulation of Igfbp-2 expression by metformin administration was disrupted by gene silencing of Ampk and Sirt1, and this phenomenon was not observed in Pparα-null mice. Notably, activation of IGF-1 receptor (IGF-1R)-dependent signaling by IGF-1 was inhibited by metformin. Finally, when compared to untreated type 2 diabetes patients, the metformin-treated diabetic patients showed increased IGFBP-2 levels with diminished serum IGF-1 levels. Taken together, these findings indicate that IGFBP-2 might be a new target of metformin action in diabetes and the metformin-AMPK-Sirt1-PPARα-IGFBP-2 network may provide a novel pathway that could be applied to ameliorate metabolic syndromes by controlling IGF-1 bioavailability.


Spare embryos: 3000 reasons to rethink the significance of genetic relatedness.
IVF laws in Victoria, Australia provide that human embryos must not remain in storage in excess of 5 years. Studies reviewing the fate of surplus human embryos reveal that close to 90% of couples choose to discard their excess embryos and that hundreds of embryos are disposed of annually. It has been argued that human embryos are a valuable resource and that there is a need to consider educational programmes to encourage couples to donate spare embryos to other infertile couples, rather than discard them. Surveys show that one reason that so few embryos are donated is that couples attach great significance to genetic parenthood. Advances in reproductive technology may necessitate a review of biological definitions of family and the importance of genetic relatedness. It can be argued that it is unreasonable to conclude that genetic ties are so significant that embryos should be discarded rather than donated and raised by non-genetically related parents. It is suggested that education programmes should encourage reflection on people's beliefs about the importance of genetic relatedness with regard to what makes a family. Open embryo donation or directed embryo donation programmes might cause couples to change their minds, or alleviate their anxiety about donating embryos to others.


A randomized trial of low-dose aspirin in the primary prevention of cardiovascular disease in women.
Randomized trials have shown that low-dose aspirin decreases the risk of a first myocardial infarction in men, with little effect on the risk of ischemic stroke. There are few similar data in women. We randomly assigned 39,876 initially healthy women 45 years of age or older to receive 100 mg of aspirin on alternate days or placebo and then monitored them for 10 years for a first major cardiovascular event (i.e., nonfatal myocardial infarction, nonfatal stroke, or death from cardiovascular causes). During follow-up, 477 major cardiovascular events were confirmed in the aspirin group, as compared with 522 in the placebo group, for a nonsignificant reduction in risk with aspirin of 9 percent (relative risk, 0.91; 95 percent confidence interval, 0.80 to 1.03; P=0.13). With regard to individual end points, there was a 17 percent reduction in the risk of stroke in the aspirin group, as compared with the placebo group (relative risk, 0.83; 95 percent confidence interval, 0.69 to 0.99; P=0.04), owing to a 24 percent reduction in the risk of ischemic stroke (relative risk, 0.76; 95 percent confidence interval, 0.63 to 0.93; P=0.009) and a nonsignificant increase in the risk of hemorrhagic stroke (relative risk, 1.24; 95 percent confidence interval, 0.82 to 1.87; P=0.31). As compared with placebo, aspirin had no significant effect on the risk of fatal or nonfatal myocardial infarction (relative risk, 1.02; 95 percent confidence interval, 0.84 to 1.25; P=0.83) or death from cardiovascular causes (relative risk, 0.95; 95 percent confidence interval, 0.74 to 1.22; P=0.68). Gastrointestinal bleeding requiring transfusion was more frequent in the aspirin group than in the placebo group (relative risk, 1.40; 95 percent confidence interval, 1.07 to 1.83; P=0.02). Subgroup analyses showed that aspirin significantly reduced the risk of major cardiovascular events, ischemic stroke, and myocardial infarction among women 65 years of age or older. In this large, primary-prevention trial among women, aspirin lowered the risk of stroke without affecting the risk of myocardial infarction or death from cardiovascular causes, leading to a nonsignificant finding with respect to the primary end point.


How do adolescents access health information? And do they ask their physicians?
To improve understanding about how high school students use electronic tools to obtain health information and how this information affects their behavior. Using a cross-sectional design, we administered an anonymous survey to high school students in grades 9 through 12 at a single private Catholic high school, inquiring about their use of electronic tools to obtain health information, topics of interest, sources used to obtain information, and modifications in their behavior based on that information. Descriptive statistics and multivariate analysis of variance were used to compare trends across grade levels. Of 705 students enrolled, 24.7% were either absent or chose not to participate in the survey. Of the remaining 531 students, 497 completed the surveys, for a response rate of 70.5% (497 of 705) and a participation rate of 93.6% (497 of 531). All students were comfortable using the Internet, and >90% used it at home and in school. Access to broadband applications averaged 95% at home and 80% at school. A significant proportion (0.66; p < 0.0001) of students reported that they trusted the information found online, and 22% (not significant) modified their behavior on the basis of the information they found. Forty-two percent searched for general health information, and 43% investigated specific medical conditions or disease states. Topics related to skin were researched significantly more than nutrition, birth control, and sexually transmitted diseases. Although a significant number of students (p < 0.05) reported conducting e-mail conversations related to health topics with their teachers, <5% had e-mail communications with physicians. These data indicate that most high school students used the Internet and broadband applications at school and at home as resources for health care information. A significant number of students trusted the online information, and at nearly one-quarter subsequently modified their behavior. Students conducted e-mail conversations with teachers about health-related topics, but few students used this tool to communicate with their physicians. This information raises questions about design and implementation of strategies to provide adolescents access to appropriate health care information, including that provided by physicians.


Matrix metalloproteinase-13 is highly expressed in destructive periodontal disease activity.
Matrix metalloproteinases (MMPs) participate in extracellular matrix degradation in physiological and pathological conditions. The available evidence suggests that MMP-13 plays a significant role in both the initiation and progress of bone resorption. The aim of our study was to identify the presence of MMP-13 in adult patients with untreated chronic periodontitis. We also determined the activity of MMP-13 present in lesions undergoing episodic attachment loss in gingival crevicular fluid (GCF) samples. After monitoring at 2 and 4 months, 21 patients showed destructive periodontitis (periodontally affected sites presenting at least two sites with > or =2 mm clinical attachment loss), and GCF samples were collected both from active and inactive sites (21 GCF samples, each). GCF was collected during a 30-second interval using a paper strip, and an immunofluorescence assay was performed to determine the basal activity of MMP-13 and the relationship between 4-aminophenylmercuric acetate (APMA)-activated total MMP-13 and basal MMP-13 activity. Gingival tissues from five patients were fixed in formalin and MMP-13 expression was demonstrated using immunohistochemistry and in situ hybridization. MMP-13 molecular forms were examined by Western immunoblotting with monoclonal antibodies. MMP-13 was found in 100% of GCF samples from patients with chronic periodontitis. Active sites, associated with tissue destruction, had significantly higher proportions of active MMP-13 and MMP-13 activity levels than their inactive counterparts (1.49 versus 1.17 ng fluorescent product, respectively; P <0.05). Western blot, immunohistochemical staining, and in situ hybridization confirmed the presence of MMP-13 in periodontal disease, with observable differences between periodontitis and healthy subjects. MMP-13 immunoreactivities were seen mainly as 55 and 48 kDa, corresponding to partially and fully activated forms, respectively, and a smaller proportion of 60-kDa proenzyme form. MMP-13 activity in GCF samples was significantly increased in active sites from progressive periodontal disease, supporting its role in the alveolar bone loss developed in this disease.


Solanum tuberosum L. cv Jayoung Epidermis Extract Inhibits Mite Antigen-Induced Atopic Dermatitis in NC/Nga Mice by Regulating the Th1/Th2 Balance and Expression of Filaggrin.
Solanum tuberosum L. cv Jayoung (JY) is a potato with dark purple flesh and contains substantial amounts of polyphenols. In this study, we evaluated the therapeutic effects of S. tuberosum L. cv JY in a mouse model of Dermatophagoides farinae body (Dfb)-induced atopic dermatitis (AD). The ethanol extract of the peel of JY (EPJ) ameliorated Dfb-induced dermatitis severity, serum levels of immunoglobulin E (IgE) and thymus and activation-regulated chemokine. Histological analysis of the skin also revealed that EPJ treatment significantly decreased mast cell infiltration. The suppression of dermatitis by EPJ treatment was accompanied by a decrease in the skin levels of type 2 helper T-cell cytokines such as interleukin (IL)-4, IL-5, and IL-13. The induction of thymic stromal lymphopoietin, which leads to a systemic Th2 response, was also decreased in the skin by EPJ. Nuclear translocation of nuclear factor-κB p65 was decreased by EPJ in Dfb-induced NC/Nga mice. The protein expression of filaggrin in the AD-like skin lesions was restored by EPJ treatment. These results suggested that EPJ may be a potential therapeutic tool for the treatment of AD.


Nonlinear osmotic properties of the cell nucleus.
In the absence of active volume regulation processes, cell volume is inversely proportional to osmolarity, as predicted by the Boyle Van't Hoff relation. In this study, we tested the hypothesis that nuclear volume has a similar relationship with extracellular osmolarity in articular chondrocytes, cells that are exposed to changes in the osmotic environment in vivo. Furthermore, we explored the mechanism of the relationships between osmolarity and nuclear size and shape. Nuclear size was quantified using two independent techniques, confocal laser scanning microscopy and angle-resolved low coherence interferometry. Nuclear volume was osmotically sensitive but this relationship was not linear, showing a decline in the osmotic sensitivity in the hypo-osmotic range. Nuclear shape was also influenced by extracellular osmolarity, becoming smoother as the osmolarity decreased. The osmotically induced changes in nuclear size paralleled the changes in nuclear shape, suggesting that shape and volume are interdependent. The osmotic sensitivity of shape and volume persisted after disruption of the actin cytoskeleton. Isolated nuclei contracted in response to physiologic changes in macromolecule concentration but not in response to physiologic changes in ion concentration, suggesting solute size has an important influence on the osmotic pressurization of the nucleus. This finding in turn implies that the diffusion barrier that causes osmotic effects is not a semi-permeable membrane, but rather due to size constraints that prevent large solute molecules from entering small spaces in the nucleus. As nuclear morphology has been associated previously with cell phenotype, these findings may provide new insight into the role of mechanical and osmotic signals in regulating cell physiology.


Resection of lymph node metastases influences survival in stage IIIC endometrial cancer.
Surgical staging of endometrial cancer identifies those patients with microscopic metastatic disease most likely to benefit from adjuvant therapy and may also confer therapeutic benefit. Our objective was to compare survival of patients who underwent resection of grossly positive lymph nodes (LN) to those with microscopically positive LN. Patients had stage IIIC endometrial cancer with pelvic and/or aortic LN metastases and underwent surgery between 1973 and 2002. Exclusion criteria included pre-surgical radiation and second primary cancer. Survival was analyzed using Kaplan-Meier method and Cox proportional hazards model. Mean age of 96 patients with stage IIIC endometrial cancer was 64. There were 45 cases with microscopic LN involvement and 51 with grossly enlarged LN. Overall, 41% had disease in aortic LN, which in 18% represented isolated aortic LN metastasis. Adjuvant therapies were given to 92% of patients (85% radiotherapy, 10% chemotherapy, 10% progestins). Among those with grossly involved LN, 86% were completely resected. Five-year disease-specific survival (DSS) was 63% in 45 patients with microscopic metastatic disease compared to 50% in 44 patients with grossly positive LN completely resected and 43% in 7 with residual macroscopic disease. In multivariable analyses, gross nodal disease not debulked (HR=6.85, P=0.009), serosal/adnexal involvement (HR=2.24, P=0.036), diagnosis prior to 1989 (HR=4.33, P<0.001), older age (HR=1.09, P<0.001), and >2 positive lymph nodes (HR=3.12, P=0.007) were associated with lower DSS. Grossly involved LN can often be completely resected in patients with stage IIIC endometrial cancer. These retrospective data provide evidence suggestive of a therapeutic benefit for lymphadenectomy in endometrial cancer.


Brain annotation toolbox: exploring the functional and genetic associations of neuroimaging results.
Advances in neuroimaging and sequencing techniques provide an unprecedented opportunity to map the function of brain regions and identify the roots of psychiatric diseases. However, the results from most neuroimaging studies, i.e. activated clusters/regions or functional connectivities between brain regions, frequently cannot be conveniently and systematically interpreted, rendering the biological meaning unclear. We describe a brain annotation toolbox that generates functional and genetic annotations for neuroimaging results. The voxel-level functional description from the Neurosynth database and gene expression profile from the Allen Human Brain Atlas are used to generate functional/genetic information for region-level neuroimaging results. The validity of the approach is demonstrated by showing that the functional and genetic annotations for specific brain regions are consistent with each other; and further the region by region functional similarity network and genetic similarity network are highly correlated for major brain atlases. One application of brain annotation toolbox is to help provide functional/genetic annotations for newly discovered regions with unknown functions, e.g. the 97 new regions identified in the Human Connectome Project. Importantly, this toolbox can help understand differences between psychiatric patients and controls, and this is demonstrated using schizophrenia and autism data, for which the functional and genetic annotations for the neuroimaging changes in patients are consistent with each other and help interpret the results. BAT is implemented as a free and open-source MATLAB toolbox and is publicly available at http://184.108.40.206:1313/post/bat. Supplementary data are available at Bioinformatics online.


Brain MRI-markers Associate Differentially with Cognitive Versus Functional Decline Leading to Dementia.
Brain MRI-markers are risk factors of dementia and decline in cognition and daily functioning. It is unknown to what extent the associations of brain MRI-markers with cognition and daily functioning are part of the pathway leading to dementia. We aimed to investigate associations of brain MRI-markers with change in cognition and daily functioning during 15 years of follow-up, including their relation to dementia. Four hundred and sixty three stroke-free and non-demented participants from the population-based Rotterdam Study that underwent brain-MRI, yielding brain volumetrics, between 1995 and 1996. We assessed cognition using the Mini-Mental State Examination (MMSE) and daily functioning using instrumental and basic activities of daily living (IADL and BADL) up to seven times between 1990 and 2011. Analyses were performed both including and excluding incident demented participants. Smaller brain volume associated with larger decline in MMSE, IADL, and BADL. Larger white matter lesion volume associated with larger decline in MMSE. Frontal lobe volume associated strongest with decline in IADL and BADL, and temporal lobe volume with decline in MMSE. After excluding incident demented participants (n = 63), associations with IADL and BADL remained, while associations with MMSE disappeared. Smaller brain volumes and larger white matter volume associate with larger decline in cognition and daily functioning, during 15 years of follow-up. Importantly, the relation of brain volume with cognition, but not daily functioning, was driven by those individuals that ultimately developed dementia.


Research support by doctoral-granting colleges/schools of nursing.
Colleges and schools of nursing with doctoral programs focus on developing quality research programs. One effective way of managing and nurturing a research program is through the implementation of a nursing research office or center. The purpose of this study is to describe the resources provided by the colleges/schools of nursing with doctoral programs for research development. A self-report questionnaire, developed by the research team, was mailed to all schools of nursing offering doctoral programs. The response rate was 79 per cent (65/82 schools). Results indicated that 56 schools (86.2 per cent) have designated research support offices. The main goals of nursing research offices are to increase the amount of extramural funding and to promote dissemination of scholarly work via publications and presentations. The majority of research offices provide assistance with grants and the research process and offer educational programs. Most doctoral-granting schools are providing some support for research activities. However, the degree of investment in research support varied widely among the responding schools. This study suggests that it takes both time and institutional commitment to build a successful research environment. Although necessary for research development, support services are not sufficient by themselves. Instead, they need to be considered in the light of individual (e.g., faculty interest and motivation) and group (e.g., culture of scholarship) factors within each school.


A trifluoromethyl analogue of celecoxib exerts beneficial effects in neuroinflammation.
Celecoxib is a selective cyclooxygenase-2 (COX2) inhibitor. We have previously shown that celecoxib inhibits experimental autoimmune encephalomyelitis (EAE) in COX-2-deficient mice, suggestive for a mode of action involving COX2-independent pathways. In the present study, we tested the effect of a trifluoromethyl analogue of celecoxib (TFM-C) with 205-fold lower COX-2 inhibitory activity in two models of neuroinflammation, i.e. cerebellar organotypic cultures challenged with LPS and the EAE mouse model for multiple sclerosis. TFM-C inhibited secretion of IL-1β, IL-12 and IL-17, enhanced that of TNF-α and RANTES, reduced neuronal axonal damage and protected from oxidative stress in the organotypic model. TFM-C blocked TNF-α release in microglial cells through a process involving intracellular retention, but induced TNF-α secretion in primary astrocyte cultures. Finally, we demonstrate that TFM-C and celecoxib ameliorated EAE with equal potency. This coincided with reduced secretion of IL-17 and IFN-γ by MOG-reactive T-cells and of IL-23 and inflammatory cytokines by bone marrow-derived dendritic cells. Our study reveals that non-coxib analogues of celecoxib may have translational value in the treatment of neuro-inflammatory conditions.


Global sea turtle conservation successes.
We document a tendency for published estimates of population size in sea turtles to be increasing rather than decreasing across the globe. To examine the population status of the seven species of sea turtle globally, we obtained 299 time series of annual nesting abundance with a total of 4417 annual estimates. The time series ranged in length from 6 to 47 years (mean, 16.2 years). When levels of abundance were summed within regional management units (RMUs) for each species, there were upward trends in 12 RMUs versus downward trends in 5 RMUs. This prevalence of more upward than downward trends was also evident in the individual time series, where we found 95 significant increases in abundance and 35 significant decreases. Adding to this encouraging news for sea turtle conservation, we show that even small sea turtle populations have the capacity to recover, that is, Allee effects appear unimportant. Positive trends in abundance are likely linked to the effective protection of eggs and nesting females, as well as reduced bycatch. However, conservation concerns remain, such as the decline in leatherback turtles in the Eastern and Western Pacific. Furthermore, we also show that, often, time series are too short to identify trends in abundance. Our findings highlight the importance of continued conservation and monitoring efforts that underpin this global conservation success story.


Gefitinib in combination with paclitaxel and carboplatin in advanced non-small-cell lung cancer: a phase III trial--INTACT 2.
Preclinical studies indicate that gefitinib (Iressa, ZD1839; AstraZeneca, Wilmington, DE), an orally active epidermal growth factor receptor tyrosine kinase inhibitor, may enhance antitumor efficacy of cytotoxics, and combination with paclitaxel and carboplatin had acceptable tolerability in a phase I trial. Gefitinib monotherapy demonstrated unparalleled antitumor activity for a biologic agent, with less toxicity than docetaxel, in phase II trials in refractory, advanced non-small-cell lung cancer (NSCLC). This phase III, randomized, placebo-controlled, double-blind trial evaluated gefitinib plus paclitaxel and carboplatin in chemotherapy-naive patients with advanced NSCLC. Patients received paclitaxel 225 mg/m(2) and carboplatin area under concentration/time curve of 6 mg/min/mL (day 1 every 3 weeks) plus gefitinib 500 mg/d, gefitinib 250 mg/d, or placebo. After a maximum of six cycles, daily gefitinib or placebo continued until disease progression. End points included overall survival, time to progression (TTP), response rate (RR), and safety evaluation. Results A total of 1,037 patients were recruited. Baseline demographic characteristics were well balanced. There was no difference in overall survival (median, 8.7, 9.8, and 9.9 months for gefitinib 500 mg/d, 250 mg/d, and placebo, respectively; P =.64), TTP, or RR between arms. Expected dose-related diarrhea and skin toxicity were observed in gefitinib-treated patients, with no new significant/unexpected safety findings from combination with chemotherapy. Subset analysis of patients with adenocarcinoma who received > or = 90 days' chemotherapy demonstrated statistically significant prolonged survival, suggesting a gefitinib maintenance effect. Gefitinib showed no added benefit in survival, TTP, or RR compared with standard chemotherapy alone. This large, placebo-controlled trial confirmed the favorable gefitinib safety profile observed in phase I and II monotherapy trials.


Elderly peoples' meals. A comparative study between elderly living in a nursing home and frail, self-managing elderly.
Sweden is one of few countries that have specific recommendations regarding mealtime habits. The importance of mealtime habits for health outcome is inadequately studied in elderly subjects. The aim of this study was to investigate the distribution of the daily meals/energy intake; the number of eating episodes per day and subjectively estimated meal-dependent variables (appetite, motivation to eat, sense of taste and sense of smell) and compare the results of elderly living in a nursing home with frail, self-managing elderly living at home. Explorative study. All analyses of energy intake were based on food records; weighed in the nursing home and estimated in the frail, self-managing elderly. Participants or contact persons estimated the subjective variables using a 10-point VAS scale. On average there were 4-5 daily eating episodes in both groups. The eating episodes were much more widespread over the day in the self-managing elderly, and their length of fasting at night was significantly shorter. The subjects estimated appetite and sense of smell was reduced in about 30-40% in both groups. Sense of taste was reduced in 40 % of the males and 10-20 % of the females. Energy intake was similar in both groups, 25 kcal/kg body weight/day, with more than a threefold variation among individual subjects. Energy intake/kg body weight correlated with a shorter length of fasting at night in the nursing home residents, however, did not correlate with ADL, number of eating episodes per day, appetite, motivation to eat, or senses of taste or smell. The self-managing elderly had more widespread eating episodes than the elderly in the nursing home, indicating that self-managing elderly exhibit larger variations in food intake preferences, however, without affecting mean energy intake. The lack of correlation between energy intake and estimated appetite, taste and smell is in line with previous findings in elderly.


Health Effects of Arsenic Longitudinal Study (HEALS): description of a multidisciplinary epidemiologic investigation.
Health Effects of Arsenic Longitudinal Study (HEALS), a multidisciplinary and large prospective cohort study in Araihazar, Bangladesh, was established to evaluate the effects of full-dose range arsenic (As) exposure on various health outcomes, including premalignant and malignant skin tumors, total mortality, pregnancy outcomes, and children's cognitive development. In this paper, we provide descriptions of the study methods including study design, study population, data collection, response rates, and exposure and outcome assessments. We also present characteristics of the study participants including the distribution of exposure and the prevalence of skin lesion at baseline recruitment. A total of 11,746 married men and women between 18 and 75 years of age participated in the study at baseline (a response rate of 98%) and completed a full questionnaire interview that included a food frequency questionnaire, with a response rate of 98%. Among the 98% of the participants who completed the clinical evaluation, over 90% provided blood samples and spot urine samples. Higher educational status, male gender, and presence of premalignant skin lesions were associated with an increased likelihood of providing blood and urine samples. Older participants were less likely to donate a blood sample. About one-third of the participants consumed water from a well with As concentration in each of three groups: >100 microg/l, 25-100 microg/l, and <25 microg/l. Average urinary As concentrations were 140 and 136 microg/l for males and females, respectively. HEALS has several unique features, including a prospective study design, comprehensive assessments of both past and future changes in As exposure at the individual level, a large repository of biological samples, and a full dose range of As exposures in the study population. HEALS is a valuable resource for examining novel research questions on the health effects of As exposure.


Non-Instrumental Movement Inhibition (NIMI) Differentially Suppresses Head and Thigh Movements during Screenic Engagement: Dependence on Interaction.
Estimating engagement levels from postural micromovements has been summarized by some researchers as: increased proximity to the screen is a marker for engagement, while increased postural movement is a signal for disengagement or negative affect. However, these findings are inconclusive: the movement hypothesis challenges other findings of dyadic interaction in humans, and experiments on the positional hypothesis diverge from it. (1) Under controlled conditions, adding a relevant visual stimulus to an auditory stimulus will preferentially result in Non-Instrumental Movement Inhibition (NIMI) of the head. (2) When instrumental movements are eliminated and computer-interaction rate is held constant, for two identically-structured stimuli, cognitive engagement (i.e., interest) will result in measurable NIMI of the body generally. Twenty-seven healthy participants were seated in front of a computer monitor and speakers. Discrete 3-min stimuli were presented with interactions mediated via a handheld trackball without any keyboard, to minimize instrumental movements of the participant's body. Music videos and audio-only music were used to test hypothesis (1). Time-sensitive, highly interactive stimuli were used to test hypothesis (2). Subjective responses were assessed via visual analog scales. The computer users' movements were quantified using video motion tracking from the lateral aspect. Repeated measures ANOVAs with Tukey post hoc comparisons were performed. For two equivalently-engaging music videos, eliminating the visual content elicited significantly increased non-instrumental movements of the head (while also decreasing subjective engagement); a highly engaging user-selected piece of favorite music led to further increased non-instrumental movement. For two comparable reading tasks, the more engaging reading significantly inhibited (42%) movement of the head and thigh; however, when a highly engaging video game was compared to the boring reading, even though the reading task and the game had similar levels of interaction (trackball clicks), only thigh movement was significantly inhibited, not head movement. NIMI can be elicited by adding a relevant visual accompaniment to an audio-only stimulus or by making a stimulus cognitively engaging. However, these results presume that all other factors are held constant, because total movement rates can be affected by cognitive engagement, instrumental movements, visual requirements, and the time-sensitivity of the stimulus.


Induction of ZEB proteins by inactivation of RB protein is key determinant of mesenchymal phenotype of breast cancer.
We previously showed that depletion of the retinoblastoma protein (RB) induces down-regulation of the adhesion molecule E-cadherin and thereby triggers the epithelial-mesenchymal transition. To further characterize the effect of RB inactivation on the phenotype of cancer cells, we have now examined RB expression in human breast cancer cell lines and clinical specimens. We found that RB-inactive cells exhibit a mesenchymal-like morphology and are highly invasive. We also found that ZEB proteins, transcriptional repressors of the E-cadherin gene, are markedly up-regulated in these cells in a manner sensitive to the miR-200 family of microRNAs. Moreover, depletion of ZEB in RB-inactive cells suppressed cell invasiveness and proliferation and induced epithelial marker expression. These results implicate ZEB in induction of the epithelial-mesenchymal transition, as well as in maintenance of the mesenchymal phenotype in RB-inactive cells. We also developed a screening program for inhibitors of ZEB1 expression and thereby identified several cyclin-dependent kinase inhibitors that blocked both ZEB1 expression and RB phosphorylation. Together, our findings suggest that RB inactivation contributes to tumor progression not only through loss of cell cycle control but also through up-regulation of ZEB expression and induction of an invasive phenotype.


Predicting behavioural intentions towards medication safety among student and new graduate nurses across four countries.
To identify final-year undergraduate students and new graduate nurses' behavioural intentions towards medication safety across four countries. Medication errors are a common and avoidable occurrence, being costly for not only patients but also for health systems and society. A multi-site cross-sectional study. A self-administered survey was distributed to students and new graduate nurses in South Africa, India, Turkey and Australia. Descriptive statistics were calculated for all survey items. Multiple linear regressions were performed to predict behavioural intentions using the three Theory of Planned Behaviour constructs: attitudes, behavioural control and subjective norms. This study adheres to the STROBE guidelines. Data were analysed for 432 students and 576 new graduate nurses. Across all countries, new graduate nurses reported significantly higher scores on all the TPB variables compared with student nurses. Attitudes towards medication management were found significantly and positively related to intention to practice safe medication management for both student and new graduate nurses. Total perceived behavioural control was significantly and negatively related to intention to practice safe medication management for students. Student and new graduate nurses showed favourable attitude, subjective norm, perceived behaviour control and intention in practising medication safety. However, differences in countries require further exploration on the factors influencing attitudes towards medication safety among student nurses and new nurse graduates. Understanding student and new graduate nurses' medication administration practices is important to inform strategies aimed at improving patient safety. The findings of this study highlight the need for an internationally coordinated approach to ensure safe medication administration by student and new graduate nurses.


Genetic testing and counseling for hereditary forms of colorectal cancer.
The discovery of genes responsible for inherited forms of colorectal cancer have the potential to improve cancer risk assessment and counseling. Germline mutations (nonsense, frameshift) of APC are associated with familial adenomatous polyposis, an autosomal dominant syndrome, clinically characterized by young onset, hundreds of adenomatous polyps in the colon, and increased risk for extracolonic tumors. Mutations in APC are also associated with forms of attenuated familial adenomatous polyposis. Germline mutations in five mismatch repair related genes (hMSH2, hMLH1, hMSH6, hPMS1, and hPMS2) cause hereditary nonpolyposis colorectal cancer and are associated with increased risk of somatic genetic alterations and high DNA microsatellite instability. Hereditary nonpolyposis colorectal cancer is characterized by young onset colorectal cancer, proximal colon location, and increased risk of extracolonic cancers. A missense mutation in APC (I1307K) is associated with some familial colorectal cancer in Ashkenazic Jews. For persons at risk for hereditary forms of colorectal cancer, testing algorithms and gene test interpretations depend on identification of the pedigree germline gene mutation. Careful evaluation of the kindred for characteristic aggregation of tumor types among affected individuals and the availability of affected persons for testing are important issues in implementing genetic testing and follow-up management. Case reports illustrate the importance of genetic counseling as a component of cancer genetic risk assessment. The genetic counseling process includes exploration of patient risk perception, sources of anxiety related to cancer risk, patient education (specific cancer-related issues, prevention/intervention options), discussion of possible gene test options, test limitations, and consequences of various gene test outcomes.


Rapid Assessment of Site Specific DNA Methylation through Resistive Pulse Sensing.
Many diseases are defined by patterns of DNA methylation which result in aberrant gene expression. We present a rapid assay based upon resistive pulse sensing, RPS, to characterize sequence specific DNA methylation sites in genomic DNA. We modify the surface of superparamagnetic beads, SPBs, with DNA (capture probe). The particles are added to solution where they bind to and extract sequence specific DNA (target DNA). The target loaded SPBs are then incubated with antibodies which bind to the methylation sites, and the velocity of the SPBs through the nanopore reveals the number and location of the epigenetic markers within the target. The approach is capable of distinguishing between different methylation sites within a DNA promoter region. Crucially the approach is not dependent on accurate sequencing of assayed DNA, with genomic regions targeted through complementary probes. As such the number of stages and reagents costs are low and the assay is complete in under 60 min which includes the incubation and run times. The format also allows simultaneous quantification of number of copies of methylated DNA, and we illustrate this with a dose response curve.


Skeletal fluorosis from the point of view of an occupational exposure to fluorides in former Czechoslovakia.
Electrolytic production of aluminium in former Czechoslovakia started in the year 1953 in the Žiar valley in the central Slovakia. However, till 1995 the hygienic conditions for health protection were not met in the factory. It underwent a reconstruction afterwards.The authors demonstrate cases of occupational skeletal fluorosis (currently rare in Europe) in 14 metallurgists which were all disclosed in foundry workers in Žiar nad Hronom as to the year 2005. The occupational disease was diagnosed after 17.7 ± 7.67 years (x±SD) of exposure in the foundry.The authors describe the clinical conditions, haematological and biochemical tests (decreased level of ionising calcium was found in serum). The content of fluorides in urine was increased (254.4±130.95 µmol/l). The average age of patients at the time of recognition of the professional etiology of the disease was 57.93±7.95 years. Eight patients were older than 60 years. Skeletal abnormalities were evaluated by using X-ray skiagraphy, estimating the Stage I-III of the skeletal fluorosis. Typically an increase of bone density was found, the compact part of long bones was coarsed, there were calcifications of the interosseous membrane between radius and ulna and some ossifications of the sacrospinal and sacrotuberous ligaments. Twelve patients suffered sensorimotor polyneuropathy of extremities, chronic bronchitis was found in 6 patients (two of them were smokers).The last occupational case was registered in the year 2001. The authors assume that aluminium production with modern technology of better safety and protection of health of workers is the key which will make the skeletal fluorosis the history in the Czech and Slovak Republic.


VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases.
The ECS (Elongin B/C-Cul2/Cul5-SOCS-box protein) complex is a member of a family of ubiquitin ligases that share a Cullin-Rbx module. SOCS-box proteins recruit substrates to the ECS complex and are linked to Cullin-Rbx via Elongin B/C. VHL has been implicated as a SOCS-box protein, but lacks a C-terminal sequence (downstream of the BC box) of the SOCS box. We now show that VHL specifically interacts with endogenous Cul2-Rbx1 in mammalian cells, whereas SOCS-box proteins associate with Cul5-Rbx2. We also identify LRR-1 and FEM1B as proteins that share a region of homology with VHL (the VHL box, including the BC box and downstream residues) and associate with Cul2-Rbx1. ECS complexes can thus be classified into two distinct protein assemblies, that is, those that contain a subunit with a VHL box (composed of the BC box and a downstream Cul2 box) that interacts with Cul2-Rbx1, and those that contain a subunit with a SOCS box (BC box and downstream Cul5 box) that interacts with Cul5-Rbx2. Domain-swapping analyses showed that the specificity of interaction of VHL-box and SOCS-box proteins with Cullin-Rbx modules is determined by the Cul2 and Cul5 boxes, respectively. Finally, RNAi-mediated knockdown of the Cul2-Rbx1 inhibited the VHL-mediated degradation of HIF-2alpha, whereas knockdown of Cul5-Rbx2 did not affect it. These data suggest that the functions of the Cul2-Rbx1 and Cul5-Rbx2 modules are distinct.


Further evidence for the non-clonal population structure of Neisseria gonorrhoeae: extensive genetic diversity within isolates of the same electrophoretic type.
Analysis of data from multilocus enzyme electrophoresis has revealed that Neisseria gonorrhoeae populations are non-clonal. Fifteen percent of 227 isolates of N. gonorrhoeae had an identical multilocus genotype (ET1) and were recovered world-wide over a 26 year period. The recovery of isolates of identical multilocus genotype from geographically and temporally unassociated hosts is a common criterion of a clonal population structure. However, in a recombining (non-clonal) population, isolates with the same multilocus genotype can arise by the random association of the most common alleles in the population. Analysis of the variation in two further enzymes, in the restriction patterns obtained from the glutamine synthetase gene, and in the DNA fragments obtained using an arbitrarily primed polymerase chain reaction, was used to show that members of ET1 were almost as variable as randomly selected N. gonorrhoeae isolates. Unlike the situation in a strongly clonal species, the 26 ET1 isolates examined were increasingly sub-divided to give 19 distinguishable groups as variation at further loci was examined, and 24 distinguishable groups when auxotypes were also considered. We conclude that, as expected of a non-clonal population, the most commonly encountered N. gonorrhoeae multilocus genotype does not define a clone.


Expression of a minus-end-directed motor protein induces Sf9 cells to form axon-like processes with uniform microtubule polarity orientation.
Neurons extend two types of processes with distinct morphologies and patterns of microtubule polarity orientation. Axons are thin cylindrical processes containing microtubules that are uniformly oriented with their plus-ends-distal to the cell body while dendrites are stout tapering processes that contain nonuniformly oriented microtubules. We have proposed that these distinct microtubule patterns are established by molecular motors that transport microtubules into each type of process with the appropriate orientation. To test the feasibility of this proposal, we have embarked on a series of studies involving the expression of vertebrate motors in insect Sf9 cells. We previously focused on a kinesin-related protein termed CHO1/MKLP1, which localizes to the midzone of the mitotic spindle, and which has been shown to have the appropriate properties to transport microtubules of opposite orientation relative to one another. Expression of a fragment of CHO1/MKLP1 containing its motor domain induces Sf9 cells to extend processes with a stout tapering morphology and a nonuniform microtubule polarity pattern similar to dendrites. Here we focus on a minus-end-directed kinesin-related motor protein termed CHO2, which localizes to the non-overlapping regions of the mitotic spindle, and which has been shown to have the appropriate properties to transport microtubules with plus-ends-leading. Sf9 cells induced to express a fragment of CHO2 containing its motor domain extend processes with a long cylindrical morphology and a uniformly plus-end-distal microtubule polarity pattern similar to axons. These results show that motor proteins have the capacity to organize distinct patterns of microtubule polarity orientation during process outgrowth, and that these patterns are intimately related to the unique morphological characteristics of the processes. Moreover, mutation of three amino acids corresponding to the ATP binding site necessary for motor function suppresses the capacity of the CHO2 fragment to induce process formation and microtubule reorganization, indicating that at least in the case of CHO2, the transport properties of the motor are essential for it to elicit these effects.


Having a lot of a good thing: multiple important group memberships as a source of self-esteem.
Membership in important social groups can promote a positive identity. We propose and test an identity resource model in which personal self-esteem is boosted by membership in additional important social groups. Belonging to multiple important group memberships predicts personal self-esteem in children (Study 1a), older adults (Study 1b), and former residents of a homeless shelter (Study 1c). Study 2 shows that the effects of multiple important group memberships on personal self-esteem are not reducible to number of interpersonal ties. Studies 3a and 3b provide longitudinal evidence that multiple important group memberships predict personal self-esteem over time. Studies 4 and 5 show that collective self-esteem mediates this effect, suggesting that membership in multiple important groups boosts personal self-esteem because people take pride in, and derive meaning from, important group memberships. Discussion focuses on when and why important group memberships act as a social resource that fuels personal self-esteem.


Visualization of Probiotic-Mediated Ca2+ Signaling in Intestinal Epithelial Cells In Vivo.
Probiotics, such as lactic acid bacteria (LAB) and Bacillus subtilis var. natto, have been shown to modulate immune responses. It is important to understand how probiotic bacteria impact intestinal epithelial cells (IECs), because IECs are the first line of defense at the mucosal surface barrier and their activities substantially affect the gut microenvironment and immunity. However, to date, their precise mechanism remains unknown due to a lack of analytical systems available for live animal models. Recently, we generated a conditional Ca2+ biosensor Yellow Cameleon (YC3.60) transgenic mouse line and established 5D (x, y, z, time, and Ca2+) intravital imaging systems of lymphoid tissues including those in Peyer's patches and bone marrow. In the present study, we further advance our intravital imaging system for intestinal tracts to visualize IEC responses against orally administrated food compounds in real time. Using this system, heat-killed B. subtilis natto, a probiotic TTCC012 strain, is shown to directly induce Ca2+ signaling in IECs in mice housed under specific pathogen-free conditions. In contrast, this activation is not observed in the Lactococcus lactis strain C60; however, when we generate germ-free YC3.60 mice and observe the LAB stimulation of IECs in the absence of gut microbiota, C60 is capable of inducing Ca2+ signaling. This is the first study to successfully visualize the direct effect of probiotics on IECs in live animals. These data strongly suggest that probiotic strains stimulate IECs under physiological conditions and that their activity is affected by the microenvironment of the small intestine, such as commensal bacteria.


Surfing amongst Oil-Tankers: Connecting Emerging Research Fields to the Current International Landscape.
The COST Action Phycomorph (FA1406) was initiated in 2015 from a handful of academic researchers, and now joins together 19 European countries and nine international partners. Phycomorph's goal is to coordinate and develop research on developmental biology in macroalgae. This is an ambitious project, as the related scientific community is small, the concepts are complex, and there is currently limited knowledge of these organisms and there are few technologies to study them. Here we report the first step in achieving this enterprise, the creation of the Phycomorph network. We share the associated strengths, pitfalls, and prospects for setting up the network in the hope that this might guide similar efforts in other fields.