Red wine polyphenol extract efficiently protects intestinal epithelial cells from inflammation via opposite modulation of JAK/STAT and Nrf2 pathways.
The development of therapeutic approaches combining efficacy and safety represents an important goal in intestinal inflammation research. Recently, evidence has supported dietary polyphenols as useful tools in the treatment and prevention of chronic inflammatory diseases, but the mechanisms of action are still poorly understood. We here reveal molecular mechanisms underlying the anti-inflammatory action of a non-alcoholic polyphenol red wine extract (RWE), operating at complementary levels via the Janus kinase/signal transducer and activator of transcription (JAK/STAT) and Nuclear factor-erythroid 2-related factor-2 (Nrf2) pathways. RWE significantly reduced the nuclear levels of phosphorylated STAT1 and also the cellular levels of phosphorylated JAK1 induced by cytokines, suppressing the JAK/STAT inflammatory signalling cascade. In turn, RWE increased the Nrf2 nuclear level, activating the Nrf2 pathway, leading not only to an up-regulation of the heme oxygenase-1 (HO-1) expression but also to an increase of the glutamate-cysteine ligase subunit catalytic (GCLc) gene expression, enhancing the GSH synthesis, thereby counteracting GSH depletion that occurs under inflammatory conditions. Overall, data indicate that the anti-inflammatory action of RWE is exerted at complementary levels, via suppression of the JAK/STAT inflammatory pathway and positive modulation of the activity of Nrf2. These results point to the potential use of the RWE as an efficient, readily available and inexpensive therapeutic strategy in the context of gastrointestinal inflammation.


On the parental origin of de novo mutation in man.
Studies tracing parental origins of human mutations by means of cytogenetic polymorphisms and RFLPs show that most trisomics arise out of maternal errors of segregation at the first meiotic division in oocytes. Temporal disturbance of meiotic progression seems likely to underly aneuploidy production in the female mouse, and this could equally be true in women, most especially as they approach the menopause when irregular cyclicity sets in. For human monosomy X, a high proportion of cases show loss of the paternal sex chromosome, and from experimental data giving similar findings in the mouse, it seems likely that the error could arise at the pronuclear stage after sperm entry into the egg, rather than at meiosis in the male. For human point mutations and structural rearrangements, a bias exists towards paternal origins. Errors arising during spermatogonial proliferation in men could contribute point mutations, these accumulating over a lifetime to give paternal age effects. For structural rearrangements, the hypersensitive stage is likely to be the post-meiotic differentiating spermatid, a stage not subject to germinal selection, and one which in Drosophila has been shown to combine high breakability with enhanced repair. Lack of a comparable cell type to the condensing spermatid of the male might be a reason why balanced structural rearrangements are produced rather rarely in females, at least in the mouse.


Association mapping reveals novel serpentine adaptation gene clusters in a population of symbiotic Mesorhizobium.
The genetic variants that underlie microbial environmental adaptation are key components of models of microbial diversification. Characterizing adaptive variants and the pangenomic context in which they evolve remains a frontier in understanding how microbial diversity is generated. The genomics of rhizobium adaptation to contrasting soil environments is ecologically and agriculturally important because these bacteria are responsible for half of all current biologically fixed nitrogen, yet they live the majority of their lives in soil. Our study uses whole-genome sequencing to describe the pan-genome of a focal clade of wild mesorhizobia that show contrasting levels of nickel adaptation despite high relatedness (99.8% identity at 16S). We observe ecotypic specialization within an otherwise genomically cohesive population, rather than finding distinct specialized bacterial lineages in contrasting soil types. This finding supports recent reports that heterogeneous environments impose selection that maintains differentiation only at a small fraction of the genome. Our work further uses a genome-wide association study to propose candidate genes for nickel adaptation. Several candidates show homology to genetic systems involved in nickel tolerance and one cluster of candidates correlates perfectly with soil origin, which validates our approach of ascribing genomic variation to adaptive divergence.


Evaluating interventions to promote patient involvement in decision-making: by what criteria should effectiveness be judged?
Interventions to inform patients about health care options and to involve them in decisions about their care are now widely advocated. The question of which criteria should be used to judge the effectiveness of such interventions has, however, received little attention. The provision of research-based information about health care effectiveness to patients and the promotion of greater patient involvement in health care decision-making are likely to have a complex range of effects on: the information provided to patients; patients' acquisition of skills; patients' knowledge and emotions; how decisions are made; the quality of decisions; professional-patient relationships; the use of health care; the health of patients; satisfaction; and the organisation and cost of health services. Opinions about which effects are most important and how they should be measured and valued will be influenced by a variety of factors, including: the rationales and motives underlying interest in patient involvement in decision-making; the forms of patient involvement envisaged; and the types of interventions being considered. In the context of health care systems which aim primarily to improve health status and well-being, health outcomes should take priority over process variables such as decision-making behaviours and patients' knowledge.


Differential specificity of endocrine FGF19 and FGF21 to FGFR1 and FGFR4 in complex with KLB.
Recent studies suggest that betaKlotho (KLB) and endocrine FGF19 and FGF21 redirect FGFR signaling to regulation of metabolic homeostasis and suppression of obesity and diabetes. However, the identity of the predominant metabolic tissue in which a major FGFR-KLB resides that critically mediates the differential actions and metabolism effects of FGF19 and FGF21 remain unclear. We determined the receptor and tissue specificity of FGF21 in comparison to FGF19 by using direct, sensitive and quantitative binding kinetics, and downstream signal transduction and expression of early response gene upon administration of FGF19 and FGF21 in mice. We found that FGF21 binds FGFR1 with much higher affinity than FGFR4 in presence of KLB; while FGF19 binds both FGFR1 and FGFR4 in presence of KLB with comparable affinity. The interaction of FGF21 with FGFR4-KLB is very weak even at high concentration and could be negligible at physiological concentration. Both FGF19 and FGF21 but not FGF1 exhibit binding affinity to KLB. The binding of FGF1 is dependent on where FGFRs are present. Both FGF19 and FGF21 are unable to displace the FGF1 binding, and conversely FGF1 cannot displace FGF19 and FGF21 binding. These results indicate that KLB is an indispensable mediator for the binding of FGF19 and FGF21 to FGFRs that is not required for FGF1. Although FGF19 can predominantly activate the responses of the liver and to a less extent the adipose tissue, FGF21 can do so significantly only in the adipose tissue and adipocytes. Among several metabolic and endocrine tissues, the response of adipose tissue to FGF21 is predominant, and can be blunted by the ablation of KLB or FGFR1. Our results indicate that unlike FGF19, FGF21 is unable to bind FGFR4-KLB complex with affinity comparable to FGFR1-KLB, and therefore, at physiological concentration less likely to directly and significantly target the liver where FGFR4-KLB predominantly resides. However, both FGF21 and FGF19 have the potential to activate responses of primarily the adipose tissue where FGFR1-KLB resides.


Radiomic Phenotypes of Mammographic Parenchymal Complexity: Toward Augmenting Breast Density in Breast Cancer Risk Assessment.
Purpose To identify phenotypes of mammographic parenchymal complexity by using radiomic features and to evaluate their associations with breast density and other breast cancer risk factors. Materials and Methods Computerized image analysis was used to quantify breast density and extract parenchymal texture features in a cross-sectional sample of women screened with digital mammography from September 1, 2012, to February 28, 2013 (n = 2029; age range, 35-75 years; mean age, 55.9 years). Unsupervised clustering was applied to identify and reproduce phenotypes of parenchymal complexity in separate training (n = 1339) and test sets (n = 690). Differences across phenotypes by age, body mass index, breast density, and estimated breast cancer risk were assessed by using Fisher exact, χ2, and Kruskal-Wallis tests. Conditional logistic regression was used to evaluate preliminary associations between the detected phenotypes and breast cancer in an independent case-control sample (76 women diagnosed with breast cancer and 158 control participants) matched on age. Results Unsupervised clustering in the screening sample identified four phenotypes with increasing parenchymal complexity that were reproducible between training and test sets (P = .001). Breast density was not strongly correlated with phenotype category (R2 = 0.24 for linear trend). The low- to intermediate-complexity phenotype (prevalence, 390 of 2029 [19%]) had the lowest proportion of dense breasts (eight of 390 [2.1%]), whereas similar proportions were observed across other phenotypes (from 140 of 291 [48.1%] in the high-complexity phenotype to 275 of 511 [53.8%] in the low-complexity phenotype). In the independent case-control sample, phenotypes showed a significant association with breast cancer (P = .001), resulting in higher discriminatory capacity when added to a model with breast density and body mass index (area under the curve, 0.84 vs 0.80; P = .03 for comparison). Conclusion Radiomic phenotypes capture mammographic parenchymal complexity beyond conventional breast density measures and established breast cancer risk factors. © RSNA, 2018 Online supplemental material is available for this article. See also the editorial by Pinker in this issue.


Maternal body mass index and spontaneous contractility of human myometrium in pregnancy.
There is controversy as to whether maternal body mass index (BMI) influences the contractility of human myometrium in pregnancy. The aim of this study was to examine spontaneous contractile activity of human pregnant myometrium in vitro, with respect to maternal BMI. Myometrial tissue specimens were obtained at cesarean delivery from 74 women with BMI values ranging from 19 to 50.1 kg m-2. By recording in vitro from eight strips per donor (590 strips in total), several parameters of spontaneous contractile activity were monitored. The relationship between BMI and contractility was evaluated using linear regression analysis. There was a significant correlation between maximum amplitude (P=0.007) and mean contractile force (P=0.001) with increasing BMI. However, the time to onset of contractions (P=0.009), and time taken to reach maximal amplitude (P=0.020) also increased with increasing BMI. No significant correlation was observed with BMI for other parameters studied. The mean maximum amplitude value for spontaneous contractions was 37±1 mN, the mean contractile force for spontaneous contractions was 4.1±0.1 mN, the average time to the first spontaneous contraction was 11.3±0.6 min and the average frequency of contractions was 6.5±0.2 per hour. These results suggest that the time to onset of contractions is increased with increasing maternal BMI, but that the force developed is greater. In all other respects, human uterine contractility is unaffected by increasing BMI. These findings underline the complexity of regulation of uterine contractility in labor with elevated maternal BMI.


Layer-by-layer growth of binary colloidal crystals.
We report the growth of binary colloidal crystals with control over the crystal orientation through a simple layer-by-layer process. Well-ordered single binary colloidal crystals with a stoichiometry of large (L) and small (S) particles of LS2 and LS were generated. In addition, we observed the formation of an LS3 superstructure. The structures formed as a result of the templating effect of the first layer and the forces exerted by the surface tension of the drying liquid. By using spheres of different composition, one component can be selectively removed, as is demonstrated in the growth of a hexagonal non-close-packed colloidal crystal.


Acute Renal Failure due to Leukaemic Infiltration in Chronic Lymphocytic Leukaemia.
Chronic lymphocytic leukemia (CLL) is a malignancy characterized by clonal proliferation and accumulation of B lymphocytes. Although leukaemic infiltration of the kidney is well recognized in CLL, acute renal failure (ARF) due to leukaemic infiltration is extremely rare. Here we present a case of ARF as the initial manifestation of CLL. The diagnosis was made by a kidney biopsy. Treatment with cyclophosphamide and prednisolone resulted in a completely improved renal function.


Disparities in tobacco cessation medication orders and fills among special populations.
There is considerable interest in measuring and eliminating health care disparities among various special populations, but there is limited understanding of their extent, causes, or potential remedies. To improve this for tobacco cessation, we measured differences in the frequency of receiving and filling cessation medication prescriptions by race, ethnicity, age, language preference, health insurance, and pregnancy. The relevant variables for all patients of a Minnesota medical group aged 18 years and older with clinician visits were extracted from the electronic medical records of 1 large medical group from March 2006 to February 2007. This was combined with claims data from 1 insurance plan that covered most of these individuals. Order and fill rates for cessation medications were then adjusted for each of the other variables. There were 32,733 current users of tobacco, 18,047 of whom had both health insurance and pharmacy claims data available. After adjustment, 15.4% overall had received an order for cessation medications during this year, but only 78% had filled it. Groups receiving fewer orders than their comparison groups were aged 18-34 years or older than 65 years, men, pregnant women, Asians and Hispanics, and those with non-English-language preference, on Medicaid, or with fewer visits. The same groups were less likely to fill that prescription, except patients with non-English preference or Medicaid. There are disparities in both the receipt of cessation medication orders and the likelihood of filling them for some special populations. The causes are likely to be complex, but this information provides a starting point for learning to improve this problem.


Urinary bacteria in adult women with urgency urinary incontinence.
This study's aims were to detect and quantify bacterial DNA in the urine of randomized trial participants about to undergo treatment for urinary urgency incontinence (UUI) without clinical evidence of urinary tract infection (UTI) and to determine if the presence of bacterial DNA in baseline urine relates to either baseline urinary symptoms or UTI risk after urinary tract instrumentation. Women without clinical evidence of baseline UTI were randomized to cystoscopic onabotulinum toxin A injection and oral placebo medication versus cystoscopic placebo injection and active oral medication. Bacterial DNA in participants' catheterized urine was measured by quantitative polymerase chain reaction (qPCR). Bacterial DNA was detected in the urine of 38.7 % of participants (60 out of 155). In these 60 qPCR-positive participants, baseline daily UUI episodes were greater than in the 95 qPCR-negative participants (5.71 [±2.60] vs 4.72 [±2.86], p = 0.004). Neither symptom severity by questionnaire nor treatment outcome was associated with qPCR status or with qPCR level in qPCR-positive subjects. In contrast, the presence of urinary bacterial DNA was associated with UTI risk: only 10 % of the qPCR-positive women developed a UTI post-treatment, while 24 % of the qPCR-negative women did so. The median qPCR level for qPCR-positive samples did not differ significantly by UTI status (UTI 2.58 × 10(5) vs no UTI 1.35 × 10(5) copies/mL, p = 0.6). These results may indicate a urinary bacterial contribution to both baseline UUI and the risk of post-treatment UTI.


Solar enhanced wastewater treatment in waste stabilization ponds.
One of the most popular off-site wastewater treatment plants used in the tropics is the waste stabilization pond (WSP). Although it has several advantages, its use in urban areas is limited because of its large land area requirement. Hence, this research is aimed at investigating if a solar-enhanced WSP (SEWSP) can increase treatment efficiency and consequently reduce the land area requirement. The SEWSPs of varying sizes, made of a metallic tank with inlet and outlet valves and a solar reflector, were constructed to increase the incident sunlight intensity. Wastewater samples collected from the inlet and outlet of the SEWSPs were examined for physio-chemical and biological characteristics for a period of 2 months. The parameters examined were total suspended solids, dissolved oxygen, 5-day biochemical oxygen demand (BOD5), chemical oxygen demand (COD), coliform, and Escherichia coli. The efficiencies of the SEWSPs, with respect to these parameters, fluctuated with temperature variation, with the shallowest SEWSP giving the highest treatment efficiency. The research revealed that the cost of treating wastewater using SEWSPs was approximately 2 times lower than the conventional WSP for the same treatment efficiencies.


In vivo autofluorescence imaging of early cancers in the human tracheobronchial tree with a spectrally optimized system.
The changes in the autofluorescence characteristics of the bronchial tissue is of crucial interest as a cancer diagnostic tool. Evidence exists that this native fluorescence or autofluorescence of bronchial tissues changes when they turn dysplastic and to carcinoma in situ. There is good agreement that the lesions display a decrease of autofluorescence in the green region of the spectrum under illumination with violet-light, and a relative increase in the red region of the spectrum is often reported. Imaging devices rely on this principle to detect early cancerous lesions in the bronchi. Based on a spectroscopic study, an industrial imaging prototype is developed to detect early cancerous lesions in collaboration with the firm Richard Wolf Endoskope GmbH, Germany. A preliminary clinical trial involving 20 patients with this spectrally optimized system shows that the autofluorescence can help to detect most lesions that would otherwise have remained invisible to an experienced endoscopist under white light illumination. A systematic off line analysis of the autofluorescence images pointed out that real-time decisional functions can be defined to reduce the number of false positive results. Using this method, a positive predictive value (PPV) of 75% is reached using autofluorescence only. Moreover, a PPV of 100% is obtained, when combining the white light (WL) mode and the autofluorescence (AF) mode, at the applied conditions. Furthermore, the sensitivity is estimated to be twice higher in the AF mode than in WL mode.


Do sex differences in the prevalence of ECG abnormalities vary across ethnic groups living in the Netherlands? A cross-sectional analysis of the population-based HELIUS study.
Major ECG abnormalities have been associated with increased risk of cardiovascular disease (CVD) burden in asymptomatic populations. However, sex differences in occurrence of major ECG abnormalities have been poorly studied, particularly across ethnic groups. The objectives were to investigate (1) sex differences in the prevalence of major and, as a secondary outcome, minor ECG abnormalities, (2) whether patterns of sex differences varied across ethnic groups, by age and (3) to what extent conventional cardiovascular risk factors contributed to observed sex differences. Cross-sectional analysis of population-based study. Multi-ethnic, population-based Healthy Life in an Urban Setting cohort, Amsterdam, the Netherlands. 8089 men and 11 369 women of Dutch, South-Asian Surinamese, African Surinamese, Ghanaian, Turkish and Moroccan origin aged 18-70 years without CVD. Age-adjusted and multivariable logistic regression analyses were performed to study sex differences in prevalence of major and, as secondary outcome, minor ECG abnormalities in the overall population, across ethnic groups and by age-groups (18-35, 36-50 and >50 years). Major and minor ECG abnormalities were less prevalent in women than men (4.6% vs 6.6% and 23.8% vs 39.8%, respectively). After adjustment for conventional risk factors, sex differences in major abnormalities were smaller in ethnic minority groups (OR ranged from 0.61 in Moroccans to 1.32 in South-Asian Surinamese) than in the Dutch (OR 0.49; 95% CI 0.36 to 0.65). Only in South-Asian Surinamese, women did not have a lower odds than men (OR 1.32; 95% CI 0.96 to 1.84). The pattern of smaller sex differences in ethnic minority groups was more pronounced in older than in younger age-groups. The prevalence of major ECG abnormalities was lower in women than men. However, sex differences were less apparent in ethnic minority groups. Conventional risk factors did not contribute substantially to observed sex differences.


Interferon-gamma binds to high and low affinity receptor components on murine macrophages.
Interferon-gamma (IFN-gamma), a glycoprotein secreted by antigen-or mitogen-activated lymphocytes, modulates the activities of lymphocytes and macrophages. For mouse macrophages, murine IFN-gamma (MuIFN-gamma) stimulates several functions, including phagocytosis, tumoricidal activity, and the increased expression of Ia and H-2 antigens of the major histocompatibility complex. Recent reports have suggested that IFN-gamma specifically binds to cell surface receptors corresponding to a single class of binding sites with a Kd of 10(-8) to 10(-10) M. We present evidence that, on the murine macrophage cell line WEHI-3, MuIFN-gamma specifically binds to two classes of binding sites with Kd of 9.1 X 10(-11) M (500 sites/cell) and 2.7 X 10(-9) M (4400 sites/cell). The higher affinity sites most likely represent the physiologically relevant receptors that mediate at least some of the actions of MuIFN-gamma.


Characterisation of the immune response to type I collagen in scleroderma.
This study was conducted to examine the frequency, phenotype, and functional profile of T lymphocytes that proliferate in response to type I collagen (CI) in patients with scleroderma (SSc). Peripheral blood mononuclear cells (PBMCs) from SSc patients, healthy controls, and rheumatoid arthritis disease controls were labeled with carboxy-fluorescein diacetate, succinimidyl ester (CFSE), cultured with or without antigen (bovine CI) for 14 days, and analysed by flow cytometry. Surface markers of proliferating cells were identified by multi-color flow cytometry. T-cell lines were derived after sorting for proliferating T cells (CFSElow). Cytokine expression in CI-responsive T cells was detected by intracellular staining/flow cytometry and by multiplex cytokine bead assay (Bio-Plex). A T-cell proliferative response to CI was detected in 8 of 25 (32%) SSc patients, but was infrequent in healthy or disease controls (3.6%; p = 0.009). The proliferating T cells expressed a CD4+, activated (CD25+), memory (CD45RO+) phenotype. Proliferation to CI did not correlate with disease duration or extent of skin involvement. T-cell lines were generated using in vitro CI stimulation to study the functional profile of these cells. Following activation of CI-reactive T cells, we detected intracellular interferon (IFN)-gamma but not interleukin (IL)-4 by flow cytometry. Supernatants from the T-cell lines generated in vitro contained IL-2, IFN-gamma, GM-CSF (granulocyte macrophage-colony-stimulating factor), and tumour necrosis factor-alpha, but little or no IL-4 and IL-10, suggesting that CI-responsive T cells express a predominantly Th1 cytokine pattern. In conclusion, circulating memory CD4 T cells that proliferate to CI are present in a subset of patients with SSc, but are infrequent in healthy or disease controls.


Ecological parameters of Lamproglena hoi (Copepoda: Lernaeidae) infection on the Bushveld smallscale yellowfish, Labeobarbus polylepis (Boulenger, 1907).
This study describes the distribution and aspects of the ecology of Lamproglena hoi. Bushveld small-scale yellowfish, Labeobarbus polylepis (Boulenger, 1907) were collected during June 2006 from the Phongolo and Assegaai rivers, March 2005 and October 2006 from the Elands River, and January 2007 and June 2008 from the Komati River in Mpumalanga, South Africa and examined for the presence of parasites. Lamproglena hoi specimens were collected from the gill filaments of the host. Specimens were fixed with warm AFA (alcohol-formaldehyde-acetic acid) and preserved in 70% ethanol. The identification of parasites took place in the laboratories of the University of Johannesburg. Twenty-five copepods (prevalence 21%, mean intensity = 4.17, abundance = 0.86) were collected on 29 fish in the Phongolo River and 46 copepods (prevalence 40%, mean intensity = 3.83, abundance = 1.53) were collected on 30 fish in the Assegaai River. One hundred and sixty eight copepods (prevalence 52%, mean intensity = 12.92, abundance = 6.72) were collected on 25 fish in 2005, and 527 copepods (prevalence 95%, mean intensity = 27.74, abundance = 26.35) were collected on 20 fish in the Elands River. One hundred and sixteen copepods (prevalence 75%, mean intensity = 7.73, abundance = 5.80) were collected on 20 fish in 2007, and 273 copepods (prevalence 63%, mean intensity = 16.06, abundance = 10.11) were collected on 27 fish in 2008 in the Komati River. Labeobarbus polylepis from these four rivers was found to have a relatively high L. hoi infection. Inseminated L. hoi females (immature) attach to the host in winter and their ovaries become conspicuous (mature). In spring fertilized eggs are stored in egg sacs hanging from the body (gravid), indicating that fertilized eggs start to hatch in spring and continued hatching into summer. Parasites prefer the median part of the second gill arch for attachment. No correlation exists between the number of parasites recorded on the gills and the sizes (total lengths) of yellowfish sampled.


The impact of lipid-based nutrient supplementation on anti-malarial antibodies in pregnant women in a randomized controlled trial.
Malaria and undernutrition frequently coexist, especially in pregnant women and young children. Nutrient supplementation of these vulnerable groups might reduce their susceptibility to malaria by improving immunity. Antibody immunity to antigens expressed by a placental-binding parasite isolate, a non-placental binding parasite isolate, merozoites and schizonts at enrolment (before 20 gestation weeks) and at 36 gestation weeks were measured in 1,009 Malawian pregnant women receiving a daily lipid-based nutrient supplement, multiple micronutrients or iron and folic acid, who were participants in a randomized clinical trial assessing the effects of nutrient supplementation on pregnancy outcomes and child development (registration ID: NCT01239693). Antibodies to placental-binding isolates significantly increased while antibodies to most merozoite antigens declined over pregnancy. Overall, after adjustment for covariates, the type of supplementation did not influence antibody levels at 36 gestation weeks or the rate of change in antibody levels from enrolment to 36 weeks. A negative association between maternal body mass index and opsonizing antibodies to placental-binding antigens (coefficient (95% CI) -1.04 (-1.84, -0.24), was observed. Similarly, women with higher socioeconomic status had significantly lower IgG and opsonizing antibodies to placental-binding antigens. Neither of these associations was significantly influenced by the supplementation type. In the current cohort nutrient supplementation did not affect anti-malarial antibody responses, but poor and undernourished mothers should be a priority group in future trials.


Permeability of selected drugs and chemicals across the blood-testis barrier of the rat.
Physiological studies by Setchell and others have described the existence of a blood-testis barrier (BTB) surrounding the seminiferous tubules of the mammalian testis. These studies were initiated to better define the role of the BTB with regard to the penetration of exogenous chemicals to male germ cells. The rete testis was cannulated in rats and fluid was collected. Test chemicals or drugs were usually administered by continuous i.v. infusion. Permeability of nonelectrolytes of various molecular sizes, acidic compounds with varying partition coefficients and pKalpha values, such as salicylic acid, barbiturates, and sulfonamides, across the BTB were studied. Permeability of nonelectrolytes was demonstrated to be dependent upon their molecular size, suggesting bulk flow through water-filled pores. On the other hand, permeability of acidic drugs with varying pKalpha values depended upon their partition coefficients. Transport of these chemicals from blood to seminferous tubules closely resembled their transport from blood to cerebrospinal fluid. It appears that the BTB is a complex multicellular system composed of membranes surrounding the semiferous tubules and the several layers of spermatogenic cells organized within the tubules, which restrict the permeability to the male germ cells of many foreign compounds. This must be borne in mind when extra-polating data from in vitro mutagenic test systems to man.


Species of Pythium from Greenhouses in Pennsylvania Exhibit Resistance to Propamocarb and Mefenoxam.
Pythium isolates from commercial greenhouses in Pennsylvania were tested in vitro and in vivo for sensitivity to propamocarb. Isolates of Pythium aphanidermatum, P. irregulare, and P. ultimum consistently infected geranium seedlings that had been treated with propamocarb. P. ultimum and P. irregulare isolates exhibited dual fungicide resistance by overcoming propamocarb, mefenoxam, and a mixture of propamocarb and mefenoxam to infect seedlings. The sensitivity of isolates to propamocarb in vitro was not a good predictor of in vivo sensitivity. This is the first report of Pythium spp. resistance to propamocarb and dual resistance to propamocarb and mefenoxam.


Reduction of Streptococcus pneumoniae Colonization and Dissemination by a Nonopsonic Capsular Polysaccharide Antibody.
Streptococcus pneumoniae colonization of the nasopharynx (NP) is a prerequisite for invasive pneumococcal disease (IPD). The marked reduction in IPD that followed the routine use of pneumococcal polysaccharide conjugate vaccines (PCVs) has been linked to reduced NP colonization with vaccine-included serotypes (STs), with the caveat that PCVs are less effective against pneumonia than against IPD. Although PCV-elicited opsonic antibodies that enhance phagocytic killing of the homologous ST are considered a key correlate of PCV-mediated protection, recent studies question this relationship for some STs, including ST3. Studies with monoclonal antibodies (MAbs) to the pneumococcal capsular polysaccharide (PPS) of ST3 (PPS3) have shown that nonopsonic, as well as opsonic, antibodies can each protect mice against pneumonia and sepsis, but the effect of these types of MAbs on NP colonization is unknown. In this study, we determined the effects of protective opsonic and nonopsonic PPS3 MAbs on ST3 NP colonization in mice. Our results show that a nonopsonic MAb reduced early NP colonization and prevented ST3 dissemination to the lungs and blood, but an opsonic MAb did not. Moreover, the opsonic MAb induced a proinflammatory NP cytokine response, but the nonopsonic MAb had an antiinflammatory effect. The effect of the nonopsonic MAb on colonization did not require its Fc region, but its antiinflammatory effect did. Our findings challenge the paradigm that opsonic MAbs are required to prevent NP colonization and suggest that further studies of the activity of nonopsonic antibodies could advance our understanding of mechanisms of PCV efficacy and provide novel correlates of protection. Pneumococcal conjugate vaccines (PCVs) have markedly reduced the incidence of invasive pneumococcal disease (IPD). Vaccine-elicited pneumococcal polysaccharide (PPS) antibodies that enhance in vitro phagocyte killing of vaccine-included serotypes (STs) (opsonic antibodies) have been considered correlates of vaccine protection and are thought to exert their effect at the initial site of infection, the nasopharynx (NP). However, the data presented here show that this is not the necessarily the case. A nonopsonic PPS monoclonal antibody (MAb) reduced pneumococcal colonization and dissemination of its homologous ST in mice, but surprisingly, an opsonic PPS MAb to the same ST did not. These results reveal that PPS antibodies can work in different ways than previously thought, challenge the paradigm that opsonic antibodies are required to prevent IPD, and provide new insights into PCV efficacy that could lead to novel correlates of vaccine protection.


Genomic DNA sequence comparison between two inbred soybean cyst nematode biotypes facilitated by massively parallel 454 micro-bead sequencing.
Heterodera glycines, the soybean cyst nematode (SCN), is a damaging agricultural pest that could be effectively managed if critical phenotypes, such as virulence and host range could be understood. While SCN is amenable to genetic analysis, lack of DNA sequence data prevents the use of such methods to study this pathogen. Fortunately, new methods of DNA sequencing that produced large amounts of data and permit whole genome comparative analyses have become available. In this study, 400 million bases of genomic DNA sequence were collected from two inbred biotypes of SCN using 454 micro-bead DNA sequencing. Comparisons to a BAC, sequenced by Sanger sequencing, showed that the micro-bead sequences could identify low and high copy number regions within the BAC. Potential single nucleotide polymorphisms (SNPs) between the two SCN biotypes were identified by comparing the two sets of sequences. Selected resequencing revealed that up to 84% of the SNPs were correct. We conclude that the quality of the micro-bead sequence data was sufficient for de novo SNP identification and should be applicable to organisms with similar genome sizes and complexities. The SNPs identified will be an important starting point in associating phenotypes with specific regions of the SCN genome.


Carcinoid Tumors of Digestive Organs: a Clinicopathologic Study of 13 Cases.
The author investigated clinicopathologic features of 13 cases of carcinoid tumor in the digestive organs. The 13 cases of carcinoid were identified from 18,267 pathological specimens of digestive organs in the last 10 years in our pathology laboratory. The tumor locations were rectum in 9 cases, duodenum in 2 cases, liver in 1 case, and stomach in 1 case. The age of the patients ranged from 52 to 82 years with a mean of 63 years. Male to female ratio was 7 : 6. The presenting symptoms were abnormal pain in 3 cases and asymptomatic in 10 cases. None of the cases showed carcinoid syndrome. The diameter ranged from 5 mm to 25 mm in gastrointestinal carcinoids, and 60 mm in the hepatic carcinoid. The treatment was endoscopic mucosal resection in 10 cases and surgical resection in 3 cases. The outcome is good except for hepatic atypical carcinoid which showed metastases and died of systemic metastasis. Histologically, 12 carcinoid tumors were typical carcinoids, and one (liver) was atypical carcinoid. Organoid pattern was present in 12 cases. Trabecular arrangement, ribbon arrangement, rosette formation, and pseudoglandular arrangement were recognized in 12 cases, in 8 cases, in 7 cases, and in 5 cases, respectively. Immunohistochemically, tumor cells were positive for at least one of pan-neuroendocrine markers including chromogranin, synaptophysin, neuron-specific enolase, CD56, and glucagon. Of these, synaptophysin was positive in 11/13 (85%), neuron-specific enolase 10/13 (80%), chromogranin 8/13 (62%), CD56 6/13 (46%), and glucagon 4/13 (31%). In summary, the author reported the incidence of digestive organ carcinoid tumors, and the clinicopathologic features of the 13 cases with carcinoid.


C9orf72, a protein associated with amyotrophic lateral sclerosis (ALS) is a guanine nucleotide exchange factor.
Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two late onset neurodegenerative diseases, have been shown to share overlapping cellular pathologies and genetic origins. Studies suggest that a hexanucleotide repeat expansion in the first intron of the C9orf72 gene is the most common cause of familial FTD and ALS pathology. The C9orf72 protein is predicted to be a differentially expressed in normal and neoplastic cells domain protein implying that C9orf72 functions as a guanine nucleotide exchange factor (GEF) to regulate specific Rab GTPases. Reported studies thus far point to a putative role for C9orf72 in lysosome biogenesis, vesicular trafficking, autophagy and mechanistic target of rapamycin complex1 (mTORC1) signaling. Here we report the expression, purification and biochemical characterization of C9orf72 protein. We conclusively show that C9orf72 is a GEF. The distinctive presence of both Rab- and Rho-GTPase GEF activities suggests that C9orf72 may function as a dual exchange factor coupling physiological functions such as cytoskeleton modulation and autophagy with endocytosis.


Adoption and eating disorders: a high-risk group?
Between 1975 and 1985, 3.8% of patients referred to the Maudsley Hospital with anorexia and bulimia were adoptees. This exceeds the rate of adoption (1.5%) in the general population matched for year of birth. The 18 adoptees with eating disorders had significantly more associated behavioural disturbance and lower academic achievement than the 18 matched controls, and different precipitating factors.


From Genetics to Functional Genomics: Improvement in Drought Signaling and Tolerance in Wheat.
Drought being a yield limiting factor has become a major threat to international food security. It is a complex trait and drought tolerance response is carried out by various genes, transcription factors (TFs), microRNAs (miRNAs), hormones, proteins, co-factors, ions, and metabolites. This complexity has limited the development of wheat cultivars for drought tolerance by classical breeding. However, attempts have been made to fill the lost genetic diversity by crossing wheat with wild wheat relatives. In recent years, several molecular markers including single nucleotide polymorphisms (SNPs) and quantitative trait loci (QTLs) associated with genes for drought signaling pathways have been reported. Screening of large wheat collections by marker assisted selection (MAS) and transformation of wheat with different genes/TFs has improved drought signaling pathways and tolerance. Several miRNAs also provide drought tolerance to wheat by regulating various TFs/genes. Emergence of OMICS techniques including transcriptomics, proteomics, metabolomics, and ionomics has helped to identify and characterize the genes, proteins, metabolites, and ions involved in drought signaling pathways. Together, all these efforts helped in understanding the complex drought tolerance mechanism. Here, we have reviewed the advances in wide hybridization, MAS, QTL mapping, miRNAs, transgenic technique, genome editing system, and above mentioned functional genomics tools for identification and utility of signaling molecules for improvement in wheat drought tolerance.


An affected core drives network integration deficits of the structural connectome in 22q11.2 deletion syndrome.
Chromosome 22q11.2 deletion syndrome (22q11DS) is a genetic disease known to lead to cerebral structural alterations, which we study using the framework of the macroscopic white-matter connectome. We create weighted connectomes of 44 patients with 22q11DS and 44 healthy controls using diffusion tensor magnetic resonance imaging, and perform a weighted graph theoretical analysis. After confirming global network integration deficits in 22q11DS (previously identified using binary connectomes), we identify the spatial distribution of regions responsible for global deficits. Next, we further characterize the dysconnectivity of the deficient regions in terms of sub-network properties, and investigate their relevance with respect to clinical profiles. We define the subset of regions with decreased nodal integration (evaluated using the closeness centrality measure) as the affected core (A-core) of the 22q11DS structural connectome. A-core regions are broadly bilaterally symmetric and consist of numerous network hubs - chiefly parietal and frontal cortical, as well as subcortical regions. Using a simulated lesion approach, we demonstrate that these core regions and their connections are particularly important to efficient network communication. Moreover, these regions are generally densely connected, but less so in 22q11DS. These specific disturbances are associated to a rerouting of shortest network paths that circumvent the A-core in 22q11DS, "de-centralizing" the network. Finally, the efficiency and mean connectivity strength of an orbito-frontal/cingulate circuit, included in the affected regions, correlate negatively with the extent of negative symptoms in 22q11DS patients, revealing the clinical relevance of present findings. The identified A-core overlaps numerous regions previously identified as affected in 22q11DS as well as in schizophrenia, which approximately 30-40% of 22q11DS patients develop.


Soluble epoxide hydrolase dimerization is required for hydrolase activity.
Soluble epoxide hydrolase (sEH) plays a key role in the metabolic conversion of the protective eicosanoid 14,15-epoxyeicosatrienoic acid to 14,15-dihydroxyeicosatrienoic acid. Accordingly, inhibition of sEH hydrolase activity has been shown to be beneficial in multiple models of cardiovascular diseases, thus identifying sEH as a valuable therapeutic target. Recently, a common human polymorphism (R287Q) was identified that reduces sEH hydrolase activity and is localized to the dimerization interface of the protein, suggesting a relationship between sEH dimerization and activity. To directly test the hypothesis that dimerization is essential for the proper function of sEH, we generated mutations within the sEH protein that would either disrupt or stabilize dimerization. We quantified the dimerization state of each mutant using a split firefly luciferase protein fragment-assisted complementation system. The hydrolase activity of each mutant was determined using a fluorescence-based substrate conversion assay. We found that mutations that disrupted dimerization also eliminated hydrolase enzymatic activity. In contrast, a mutation that stabilized dimerization restored hydrolase activity. Finally, we investigated the kinetics of sEH dimerization and found that the human R287Q polymorphism was metastable and capable of swapping dimer partners faster than the WT enzyme. These results indicate that dimerization is required for sEH hydrolase activity. Disrupting sEH dimerization may therefore serve as a novel therapeutic strategy for reducing sEH hydrolase activity.


High-dose steroid treatment increases free water transport in peritoneal dialysis patients.
The water channel aquaporin-1 (AQP1) is the molecular counterpart of the ultrasmall pore that mediates free water transport during peritoneal dialysis (PD). Proof-of-principle studies performed in rats have shown that treatment with corticosteroids upregulates the expression of AQP1 in the peritoneal capillaries, causing a significant increase in free water transport. Whether such a beneficial effect could be observed in end-stage renal disease patients treated by PD remains unknown. Peritoneal transport parameters were evaluated in three patients on PD, shortly before and after living-donor renal transplantation and treatment with high-dose methylprednisolone (1.0-1.2 g/m(2)). As compared with pre-transplantation values, the post-transplantation test revealed an ∼2-fold increase in the sodium sieving and ultrasmall pore ultrafiltration volume, suggesting an effect on AQP1 water channels. In contrast, there was no change in the parameters of small solute transport. The direct involvement of AQP1 in these changes is suggested by the expression of glucocorticoid receptors in the human peritoneum and the presence of conserved glucocorticoid response elements in the promoter of the human AQP1 gene.


NKG2D triggers cytotoxicity in mouse NK cells lacking DAP12 or Syk family kinases.
In activated mouse natural killer (NK) cells, the NKG2D receptor associates with two intracellular adaptors, DAP10 and DAP12, which trigger phosphatidyl inositol 3 kinase (PI3K) and Syk family protein tyrosine kinases, respectively. Here we show that cytotoxicity, but not cytokine production, is triggered by NKG2D in activated NK cells lacking either DAP12 or the Syk family members Syk and ZAP70. Inhibition of PI3K blocks this cytotoxicity, suggesting that the DAP10-PI3K pathway is sufficient to initiate NKG2D-mediated killing of target cells. Our results highlight signaling divergence in the effector functions of NKG2D and indicate that alternative associations between a receptor and its adaptors may provide a single receptor with a dual 'on-switch', giving mouse NK cells more choices through which to trigger cytotoxicity.


Interactions between breast cancer susceptibility loci and menopausal hormone therapy in relationship to breast cancer in the Breast and Prostate Cancer Cohort Consortium.
Current use of menopausal hormone therapy (MHT) has important implications for postmenopausal breast cancer risk, and observed associations might be modified by known breast cancer susceptibility loci. To provide the most comprehensive assessment of interactions of prospectively collected data on MHT and 17 confirmed susceptibility loci with invasive breast cancer risk, a nested case-control design among eight cohorts within the NCI Breast and Prostate Cancer Cohort Consortium was used. Based on data from 13,304 cases and 15,622 controls, multivariable-adjusted logistic regression analyses were used to estimate odds ratios (OR) and 95 % confidence intervals (CI). Effect modification of current and past use was evaluated on the multiplicative scale. P values <1.5 × 10(-3) were considered statistically significant. The strongest evidence of effect modification was observed for current MHT by 9q31-rs865686. Compared to never users of MHT with the rs865686 GG genotype, the association between current MHT use and breast cancer risk for the TT genotype (OR 1.79, 95 % CI 1.43-2.24; P interaction = 1.2 × 10(-4)) was less than expected on the multiplicative scale. There are no biological implications of the sub-multiplicative interaction between MHT and rs865686. Menopausal hormone therapy is unlikely to have a strong interaction with the common genetic variants associated with invasive breast cancer.


An analysis of methylenetetrahydrofolate reductase and glutathione S-transferase omega-1 genes as modifiers of the cerebral response to ischemia.
Cerebral ischemia involves a series of reactions which ultimately influence the final volume of a brain infarction. We hypothesize that polymorphisms in genes encoding proteins involved in these reactions could act as modifiers of the cerebral response to ischemia and impact the resultant stroke volume. The final volume of a cerebral infarct is important as it correlates with the morbidity and mortality associated with non-lacunar ischemic strokes. The proteins encoded by the methylenetetrahydrofolate reductase (MTHFR) and glutathione S-transferase omega-1 (GSTO-1) genes are, through oxidative mechanisms, key participants in the cerebral response to ischemia. On the basis of these biological activities, they were selected as candidate genes for further investigation. We analyzed the C677T polymorphism in the MTHFR gene and the C419A polymorphism in the GSTO-1 gene in 128 patients with non-lacunar ischemic strokes. We found no significant association of either the MTHFR (p = 0.72) or GSTO-1 (p = 0.58) polymorphisms with cerebral infarct volume. Our study shows no major gene effect of either the MTHFR or GSTO-1 genes as a modifier of ischemic stroke volume. However, given the relatively small sample size, a minor gene effect is not excluded by this investigation.


A screening-level approach to quantifying risk from glacial release of organochlorine pollutants in the Alaskan Arctic.
Widespread distribution of atmospherically mobilized organochlorine pollutants (OCPs) has been documented throughout the Arctic. A fraction of these OCPs have become entrained in glacial ice, and during melting, they can be released into downstream reservoirs. Though this remobilization is known, an assessment of risk from glacial meltwater to collocated human communities in the Arctic, including Alaska, had not been accomplished. Here, we use a screening-level risk assessment model for glacial watersheds, based on US Environmental Protection Agency (EPA) methodology, which we apply to the glaciated Jarvis Creek watershed of interior Alaska. Model results indicate that even with low levels of OCPs in glacial meltwater, high fish consumption by subsistence communities in the area increases the risk of cancer and hazard impacts above acceptable limits. Though this model is specific to one watershed, our results imply that further investigation of an increasing OCP signal in glacial meltwater and fish throughout the North American Arctic is warranted.


What are the limitations on the wider therapeutic use of phage?
Bacterial resistance to antibiotics poses a serious health threat. Since research into new antibiotics is not progressing at the same rate as the development of bacterial resistance, widespread calls for alternatives to antibiotics have been made. Phage therapy is an ideal alternative candidate to be investigated. However the success of phage therapy may be hampered by a lack of investment support from large pharmaceutical companies, due to their narrow spectrum of activity in antibiotics, very large costs associated with clinical trials of the variety of phages needed, and regulatory requirements remaining unclear. Intellectual property is difficult to secure for therapeutic phage products for a variety of reasons, and patenting procedures vary widely between the US and the EU. Consequently, companies are more likely to invest in phage products for decontamination or veterinary use, rather than clinical use in humans. Some still raise questions as to the safety of phage therapy overall, suggesting the possibility of cytotoxicity and immunogenicity, depending on the phage preparation and route. On the other hand, with patients dying because of infections untreatable with conventional antibiotics, the question arises as to whether it is ethical not to pursue phage therapy more diligently. A paradigm shift about how phage therapy is perceived is required, as well as more rigorous proof of efficacy in the form of clinical trials of existing medicinal phage products. Phage therapy potential may be fulfilled in the meantime by allowing individual preparations to be used on a named-patient basis, with extensive monitoring and multidisciplinary team input. The National Health Service and academia have a role in carrying out clinical phage research, which would be beneficial to public health, but not necessarily financially rewarding.


Fluid dynamics during bleb formation in migrating cells in vivo.
Blebs are cellular protrusions observed in migrating cells and in cells undergoing spreading, cytokinesis, and apoptosis. Here we investigate the flow of cytoplasm during bleb formation and the concurrent changes in cell volume using zebrafish primordial germ cells (PGCs) as an in vivo model. We show that bleb inflation occurs concomitantly with cytoplasmic inflow into it and that during this process the total cell volume does not change. We thus show that bleb formation in primordial germ cells results primarily from redistribution of material within the cell rather than being driven by flow of water from an external source.


Using Varying Diagnostic Criteria to Examine Mild Cognitive Impairment Prevalence and Predict Dementia Incidence in a Community-Based Sample.
Lack of a unitary operational definition of mild cognitive impairment (MCI) has resulted in mixed prevalence rates and unclear predictive validity regarding conversion to dementia and likelihood of reversion. We examined 1,721 nondemented participants aged 65 and older from the Adult Changes in Thought (ACT) community-based cohort. Participants were followed longitudinally through biennial visits (average years assessed = 5.38). Categorization of MCI was based on: 1) deviation of neuropsychological test scores from a benchmark based on either standard or individualized expectations of a participant's mean premorbid cognitive ability, and 2) cutoff for impairment (1.0 versus 1.5 standard deviations [sd] below benchmark). MCI prevalence ranged from 56-92%; using individualized benchmarks and less stringent cutoffs produced higher rates. During follow-up, 17% of the cohort developed dementia. Examination of sensitivity, specificity, and predictive validity revealed that the criterion of 1.5 sd from the standardized benchmark was optimal, but still had limited predictive validity. Participants meeting this criterion at their first visit were three times more likely to develop dementia and this increased to seven times if participants had this diagnosis at the second timepoint as well. Those who did not have an MCI diagnosis at their first visit, but did at their second, had a significant increase of risk (but to a lesser extent than those diagnosed at both visits), while those who had an MCI diagnosis at their first visit, but not their second, did not have a significantly increased risk. These results highlight how assessing MCI stability greatly improves prediction of risk.


A photochromic agonist for μ-opioid receptors.
Opioid receptors (ORs) are widely distributed in the brain, the spinal cord, and the digestive tract and play an important role in nociception. All known ORs are G-protein-coupled receptors (GPCRs) of family A. Another well-known member of this family, rhodopsin, is activated by light through the cis/trans isomerization of a covalently bound chromophore, retinal. We now show how an OR can be combined with a synthetic azobenzene photoswitch to gain light sensitivity. Our work extends the reach of photopharmacology and outlines a general strategy for converting Family A GPCRs, which account for the majority of drug targets, into photoreceptors.


Effects of Dairy Products Consumption on Health: Benefits and Beliefs--A Commentary from the Belgian Bone Club and the European Society for Clinical and Economic Aspects of Osteoporosis, Osteoarthritis and Musculoskeletal Diseases.
Dairy products provide a package of essential nutrients that is difficult to obtain in low-dairy or dairy-free diets, and for many people it is not possible to achieve recommended daily calcium intakes with a dairy-free diet. Despite the established benefits for bone health, some people avoid dairy in their diet due to beliefs that dairy may be detrimental to health, especially in those with weight management issues, lactose intolerance, osteoarthritis, rheumatoid arthritis, or trying to avoid cardiovascular disease. This review provides information for health professionals to enable them to help their patients make informed decisions about consuming dairy products as part of a balanced diet. There may be a weak association between dairy consumption and a possible small weight reduction, with decreases in fat mass and waist circumference and increases in lean body mass. Lactose intolerant individuals may not need to completely eliminate dairy products from their diet, as both yogurt and hard cheese are well tolerated. Among people with arthritis, there is no evidence for a benefit to avoid dairy consumption. Dairy products do not increase the risk of cardiovascular disease, particularly if low fat. Intake of up to three servings of dairy products per day appears to be safe and may confer a favourable benefit with regard to bone health.


H.U.B city steps: methods and early findings from a community-based participatory research trial to reduce blood pressure among African Americans.
Community-based participatory research (CBPR) has been recognized as an important approach to develop and execute health interventions among marginalized populations, and a key strategy to translate research into practice to help reduce health disparities. Despite growing interest in the CBPR approach, CBPR initiatives rarely use experimental or other rigorous research designs to evaluate health outcomes. This behavioral study describes the conceptual frameworks, methods, and early findings related to the reach, adoption, implementation, and effectiveness on primary blood pressure outcomes. The CBPR, social support, and motivational interviewing frameworks are applied to test treatment effects of a two-phased CBPR walking intervention, including a 6-month active intervention quasi experimental phase and 12-month maintenance randomized controlled trial phase to test dose effects of motivational interviewing. A community advisory board helped develop and execute the culturally-appropriate intervention components which included social support walking groups led by peer coaches, pedometer diary self-monitoring, monthly diet and physical activity education sessions, and individualized motivational interviewing sessions. Although the study is on-going, three month data is available and reported. Analyses include descriptive statistics and paired t tests. Of 269 enrolled participants, most were African American (94%) females (85%) with a mean age of 43.8 (SD = 12.1) years. Across the 3 months, 90% of all possible pedometer diaries were submitted. Attendance at the monthly education sessions was approximately 33%. At the 3-month follow-up 227 (84%) participants were retained. From baseline to 3-months, systolic BP [126.0 (SD = 19.1) to 120.3 (SD = 17.9) mmHg; p < 0.001] and diastolic BP [83. 2 (SD = 12.3) to 80.2 (SD = 11.6) mmHg; p < 0.001] were significantly reduced. This CBPR study highlights implementation factors and signifies the community's active participation in the development and execution of this study. Reach and representativeness of enrolled participants are discussed. Adherence to pedometer diary self-monitoring was better than education session participation. Significant decreases in the primary blood pressure outcomes demonstrate early effectiveness. Importantly, future analyses will evaluate long-term effectiveness of this CBPR behavioral intervention on health outcomes, and help inform the translational capabilities of CBPR efforts.


Characterizing ovarian gene expression during oocyte growth in Atlantic cod (Gadus morhua).
Vertebrate oocytes undergo dramatic changes during development as they accumulate many RNA transcripts, glycoproteins, and yolk proteins, necessary to ensure proper fertilization and embryogenesis. Oogenesis in teleosts often requires several years for completion, but very little is known about the early developmental stages. Recently, two-stage gene expression comparisons were made during oocyte growth in coho salmon (Oncorhynchus kisutch) and Atlantic cod (Gadus morhua), but more broad-scale, comprehensive assessments have not been conducted. The objectives of the present study were to characterize the gene expression patterns throughout oocyte growth in cod and compare them to changes previously identified in coho salmon. A quantitative PCR survey was conducted using 50 genes at six ovarian stages, ranging from the onset of primary growth (oocyte differentiation) to late vitellogenesis. Most expression patterns could be grouped into three major clusters, consisting of oocyte-derived (cluster 1) and likely follicle cell (clusters 2 and 3) genes. Oocyte genes were elevated during primary growth, while many follicle cell transcripts were abundant during oocyte differentiation and vitellogenesis. Few expression changes identified in coho salmon were evident in cod, which is likely due to differences in reproductive strategies. These results demonstrate that dynamic changes in gene expression occur during oocyte growth in teleost fish.


Catalytic mechanism of human DNA polymerase lambda with Mg2+ and Mn2+ from ab initio quantum mechanical/molecular mechanical studies.
DNA polymerases play a crucial role in the cell cycle due to their involvement in genome replication and repair. Understanding the reaction mechanism by which these polymerases carry out their function can provide insights into these processes. Recently, the crystal structures of human DNA polymerase lambda (Pollambda) have been reported both for pre- and post-catalytic complexes [García-Díaz et al., DNA Repair 3 (2007), 1333]. Here we employ the pre-catalytic complex as a starting structure for the determination of the catalytic mechanism of Pollambda using ab initio quantum mechanical/molecular mechanical methods. The reaction path has been calculated using Mg(2+) and Mn(2+) as the catalytic metals. In both cases the reaction proceeds through a two-step mechanism where the 3'-OH of the primer sugar ring is deprotonated by one of the conserved Asp residues (D490) in the active site before the incorporation of the nucleotide to the nascent DNA chain. A significant charge transfer is observed between both metals and some residues in the active site as the reaction proceeds. The optimized reactant and product structures agree with the reported crystal structures. In addition, the calculated reaction barriers for both metals are close to experimentally estimated barriers. Energy decomposition analysis to explain individual residue contributions suggests that several amino acids surrounding the active site are important for catalysis. Some of these residues, including R420, R488 and E529, have been implicated in catalysis by previous mutagenesis experiments on the homologous residues on Polbeta. Furthermore, Pollambda residues R420 and E529 found to be important from the energy decomposition analysis, are homologous to residues R183 and E295 in Polbeta, both of which are linked to cancer. In addition, residues R386, E391, K422 and K472 appear to have an important role in catalysis and could be a potential target for mutagenesis experiments. There is partial conservation of these residues across the Pol X family of DNA polymerases.


Structural and computational studies of the Staphylococcus aureus sortase B-substrate complex reveal a substrate-stabilized oxyanion hole.
Sortase cysteine transpeptidases covalently attach proteins to the bacterial cell wall or assemble fiber-like pili that promote bacterial adhesion. Members of this enzyme superfamily are widely distributed in Gram-positive bacteria that frequently utilize multiple sortases to elaborate their peptidoglycan. Sortases catalyze transpeptidation using a conserved active site His-Cys-Arg triad that joins a sorting signal located at the C terminus of their protein substrate to an amino nucleophile located on the cell surface. However, despite extensive study, the catalytic mechanism and molecular basis of substrate recognition remains poorly understood. Here we report the crystal structure of the Staphylococcus aureus sortase B enzyme in a covalent complex with an analog of its NPQTN sorting signal substrate, revealing the structural basis through which it displays the IsdC protein involved in heme-iron scavenging from human hemoglobin. The results of computational modeling, molecular dynamics simulations, and targeted amino acid mutagenesis indicate that the backbone amide of Glu(224) and the side chain of Arg(233) form an oxyanion hole in sortase B that stabilizes high energy tetrahedral catalytic intermediates. Surprisingly, a highly conserved threonine residue within the bound sorting signal substrate facilitates construction of the oxyanion hole by stabilizing the position of the active site arginine residue via hydrogen bonding. Molecular dynamics simulations and primary sequence conservation suggest that the sorting signal-stabilized oxyanion hole is a universal feature of enzymes within the sortase superfamily.


Relationship between erectile dysfunction and silent coronary artery disease: detection with multidetector computed tomography coronary angiography.
Our aim was to determine the relationship between erectile dysfunction (ED) and silent coronary artery disease (CAD) by multidetector computed tomography (MDCT) coronary angiography. Thirty consecutive men with nonhormonal and nonpsychogenic ED and with no cardiac symptoms were evaluated. Medical history, physical examination and laboratory investigation were performed. The five-item brief form of the International Index of Erectile Function (IIEF-5) was performed for evaluation of ED. The Agatston score (AS) was determined from MDCT images under beta blockade to induce bradycardia. The MDCT coronary angiography findings were evaluated by two radiologists blinded to the clinical findings. Patients were classified into three categories (mild, moderate and severe ED) according to IIEF-5 scores and into five categories (very low, low, moderate, moderately high and high CAD risk) according to the AS. Mean age was 58.3 ± 8.7 years (46-79). 6 patients had hypertriglyceridemia, 4 had hypercholesterolemia and 4 had hyperglycemia. All patients had normal early morning testosterone levels. Regarding IIEF-5 scores, none of them had mild ED, 14 had moderate ED and 16 had severe ED. Of the 14 patients with moderate ED, 21.4% had low and 28.5% had moderate CAD risk regarding AS. Of the 16 patients with severe ED, 25% had moderate, 31.2% had moderately high and 25% had high CAD risk regarding AS. Increasing age was a risk factor for high AS (p = 0.045). There was a significant correlation between AS and ED severity (p = 0.01). ED and CAD often coexist. MDCT coronary angiography can detect coronary lesions and allow appropriate medical intervention.


Light-triggered C60 release from a graphene/cyclodextrin nanoplatform for the protection of cytotoxicity induced by nitric oxide.
An ultraviolet (UV) light-triggered nanocarbon hybrid is developed for controlled C60 release with excellent nitric oxide (NO) quenching ability. This nanocarrier, consisting of reduced graphene oxide (rGO) and β-cyclodextrin (β-CD), is capable of hosting azobenzene functionalized C60 (Azo-C60) synthesized by diazo chemistry. The hybridization of rGO, β-CD and Azo-C60 enhances cellular uptake and limits the aggregation of C60, and shows enhanced protective effects on NO-induced cytotoxicity. More interestingly, azo groups can reversibly switch between trans- and cis-isomers upon UV irradiation, so that the Azo-C60 molecules exhibit photo-controlled release from rGO/β-CD in living cells. In vitro studies show that rGO/β-CD/C60 treated with UV irradiation causes higher NO scavenging efficacy, which further significantly increases the cell viability from 32.6% to 88.4% at low loading levels (50 μg mL-1). This represents an excellent NO quenching efficiency, better than other reports of the graphene/C60 nanohybrids, and indicates that this material can be an effective nanoplatform to combat oxidative damage. As the host-guest chemistry and diazo chemistry are versatile and universally applicable, it is worth noting that the present strategy can also be applied in preparing other photo-responsive nanohybrids, which should be valuable for use in life science and materials science.


[Risk factors and frequency of positive antibodies for leptospirosis in a sub urban population near Santiago].
Leptospirosis is a zoonotic disease and its incidence is known in Chile since 2002, when it was incorporated as a disease that must be reported to health authorities. A serologic survey for leptospirosis was performed in humans and animals from a farm in a semi urban area in Santiago Chile, after the death of a farmer due to Weil disease in that place. To report the prevalence of antibodies against leptospirosis and to determine exposure to infection risk factors in the humans and domestic animals studied in this survey. Antibodies were detected by IgM immunodot and indirect haemagglutination test in 61 humans (43 male, aged 5 to 70 years) and by microscopic agglutination test (MAT) in 44 animals. A questionnaire was applied to determine their exposure to risk factors for infection with Leptospira. Seventy two percent of the studied population were farm workers and 70% had activities that required contact with water from canals, 41% cleaned closed places where rodents were present. Other risk factors detected were lack of sewage and waste disposal, high level of rodent infestation and disposal of faeces into canals used for watering. Two humans (3.3%) and six animals (1 bovine and 5 rodents) had positive antibodies. Among animals, antibodies against Leptospira serovar pomona and icterohaemorragiae were detected. In Chile, leptospirosis exists not only in rural areas but semi urban ones close to Santiago, although the prevalence is low. Education is necessary among semi urban population to avoid infection.


Therapeutic use of humor in occupational therapy.
Interviews with five occupational therapists who use humor therapeutically in their practice were conducted and analyzed with a phenomenological method so that the lived experience of therapeutic humor use in occupational therapy could be examined. Sixteen themes were identified through data analysis: The Concept of Therapeutic Use of Humor; Spontaneous Versus Deliberate Humor; Humor, the Great Equalizer; Humor and Professionalism; Contraindications of Humor; Humor Among Co-Workers; Humor and Play; Humor and the Environment; Humor Providing Balance; The Intrinsic Quality of Humor; The Transformative Power of Humor; The Effects of Humor on the Subjects Themselves; Humor as an Evaluation and Treatment Tool; Humor as Therapeutic Use of Self; Humor as a Coping Mechanism; and Other Uses of Humor With Patients. This study revealed that the use of therapeutic humor in occupational therapy is a multifaceted phenomenon, much richer than had been previously presented in the literature.


Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Ralstonia eutropha in high cell density palm oil fermentations.
Improved production costs will accelerate commercialization of polyhydroxyalkanoate (PHA) polymer and PHA-based products. Plant oils are considered favorable feedstocks, due to their high carbon content and relatively low price compared to sugars and other refined carbon feedstocks. Different PHA production strategies were compared using a recombinant strain of Ralstonia eutropha that produces high amounts of P(HB-co-HHx) when grown on plant oils. This R. eutropha strain was grown to high cell densities using batch, extended batch, and fed batch fermentation strategies, in which PHA accumulation was triggered by nitrogen limitation. While extended batch culture produced more biomass and PHA than batch culture, fed batch cultivation was shown to produce the highest levels of biomass and PHA. The highest titer achieved was over 139 g/L cell dry weight (CDW) of biomass with 74% of CDW as PHA containing 19 mol% HHx. Our data suggest that the fermentation process is scalable with a space time yield (STY) better than 1 g PHA/L/h. The achieved biomass concentration and PHA yield are among the highest reported for the fermentation of recombinant R. eutropha strains producing P(HB-co-HHx).


Life stress and psychosomatic symptoms among 14 to 16-year old Finnish adolescents.
School children (N = 2013) completed questionnaires in class on three occasions over a period of 17 months. Life stress data for 12 months were gathered retrospectively in the last questionnaire. Life events and interpersonal problems were associated with psychosomatic symptoms. In terms of change in symptoms, the impact of life events and interpersonal problems was demonstrated in boys but not in girls. Though boys with a history of no events or problems had lower symptom scores than girls, boys with a history of many events or problems caught up with the girls by the end of the follow-up period. On the other hand, symptom differences related to life stress scores were already apparent at the beginning of the study, and these differences were greater in girls than in boys.


Ultrasensitive detection enabled by nonlinear magnetization of nanomagnetic labels.
Geometrically confined magnetic particles due to their unique response to external magnetic fields find a variety of applications, including magnetic guidance, heat and drug delivery, magneto-mechanical actuation, and contrast enhancement. Highly sensitive detection and imaging techniques based on the nonlinear properties of nanomagnets were recently proposed as innovative strong-translational potential methods applicable in complex, often opaque, biological systems. Here we report on the significant enhancement of the detection capability using optical-lithography-defined, ferromagnetic iron-nickel alloy disk-shaped particles. We show that an irreversible transition between strongly non-collinear (vortex) and single domain states, driven by an alternating magnetic field, translates into a nonlinear magnetic response that enables ultrasensitive detection of these particles. The record sensitivity of ∼3.5 × 10-9 emu, which is equivalent to ∼39 pg of magnetic material is demonstrated at room temperature for arrays of patterned disks. We also show that unbound disks suspended in the aqueous buffer can be successfully detected and quantified in real-time when administered into a live animal allowing for tracing of their biodistribution. The use of nanoscale ferromagnetic particles with engineered nonlinear properties opens prospects for further enhancing the sensitivity, scalability, and tunability of noise-free magnetic tag detection in high-background environments for various applications spanning from biosensing and medical imaging to anti-counterfeiting technologies.


Exposure to the Chinese famine in early life and the risk of hyperglycemia and type 2 diabetes in adulthood.
Early developmental adaptations in response to undernutrition may play an essential role in susceptibility to type 2 diabetes, particularly for those experiencing a "mismatched rich nutritional environment" in later life. We examined the associations of exposure to the Chinese famine (1959-1961) during fetal life and childhood with the risk of hyperglycemia and type 2 diabetes in adulthood. We used the data for 7,874 rural adults born between 1954 and 1964 in selected communities from the cross-sectional 2002 China National Nutrition and Health Survey. Hyperglycemia was defined as fasting plasma glucose ≥6.1 mmol/l and/or 2-h plasma glucose ≥7.8 mmol/l and/or a previous clinical diagnosis of type 2 diabetes. Prevalences of hyperglycemia among adults in nonexposed, fetal exposed, early-childhood, mid-childhood, and late-childhood exposed cohorts were 2.4%, 5.7%, 3.9%, 3.4%, and 5.9%, respectively. In severely affected famine areas, fetal-exposed subjects had an increased risk of hyperglycemia compared with nonexposed subjects (odds ratio = 3.92; 95% CI: 1.64-9.39; P = 0.002); this difference was not observed in less severely affected famine areas (odds ratio = 0.57; 95% CI: 0.25-1.31; P = 0.185). The odds ratios were significantly different between groups from the severe and less severe famine areas (P for interaction = 0.001). In severely affected famine areas, fetal-exposed subjects who followed an affluent/Western dietary pattern (odds ratios = 7.63; 95% CI: 2.41-24.1; P = 0.0005) or who had a higher economic status in later life experienced a substantially elevated risk of hyperglycemia (odds ratios = 6.20; 95% CI: 2.08-18.5; P = 0.001). Fetal exposure to the severe Chinese famine increases the risk of hyperglycemia in adulthood. This association appears to be exacerbated by a nutritionally rich environment in later life.


Role of FcαR EC2 region in extracellular membrane localization.
The FcαR receptor (CD89) binds to the constant region of Immunoglobulin (Ig) A to mediate mucosal immunity [1-2]. FcαR consist of five exons: two that code for the signal peptide regions S1 & S2, two for the extracellular regions EC1 and EC2, and the final exon for the transmembrane/cytoplasmic tail region [3]. Previously, we reported that the EC1 region plays an essential role for extracellular membrane localization of the receptor [4], where the absence of EC1 would prevent the variants from localizing to the cell surface, even with a full signal peptide. In the case of FcαR Variant 4 (lacking the S2 region only), there was some "leakiness" to membrane surface localization.


Fine structure of the origin-proximal DNAase I-hypersensitive region in wild-type and EC mutant polyoma.
The chromatin of wild-type polyoma virus displays a unique DNAase I highly sensitive region in situ in the infected nuclei, extending for about 260 nucleotides from the origin of replication to the beginning of the late region. We show that this highly sensitive region is not homogeneous. It displays a well defined pattern of differential sensitivity along its 260 nucleotides, including one protected subregion and two hypersensitive sites, which concern a unique residue or very few nucleotides. All these features were mapped to a precision of +/- 5 bp relative to the DNA nucleotide sequence. In parallel, we studied a PyEC mutant, whose sequence is grossly rear-ranged in this very region. This allows the PyEC to overcome the block in the expression of the early genes in the mouse embryonal carcinoma PCC4 cell line. We show that this mutant, however, displays an identical highly sensitive region with the same fine structure. The mapping of this structure on the mutant nucleotides sequence coincides with that of the wild-type relative to any arbitrary point on the late side of the rearrangement; however, 60% of the mutant molecules display this unique local chromatin structure, instead of 20% for the wild-type ones. Finally, the sequence determinism of this singularity is discussed, as well as its possible role in the control of the early transcription and the establishment of the PyEC phenotype..


A role for FoxN3 in the development of cranial cartilages and muscles in Xenopus laevis (Amphibia: Anura: Pipidae) with special emphasis on the novel rostral cartilages.
The origin of morphological novelties is a controversial topic in evolutionary developmental biology. The heads of anuran larvae have several unique structures, including the supra- and infrarostral cartilages, the specialised structure of the gill basket (used for filtration), and novel cranial muscle arrangements. FoxN3, a member of the forkhead/winged helix family of transcription factors, has been implicated as important for normal craniofacial development in the pipid anuran Xenopus laevis. We have investigated the effects of functional knockdown of FoxN3 (using antisense oligonucleotide morpholino) on the development of the larval head skeleton and the associated cranial muscles in X. laevis. Our data complement earlier studies and provide a more complete account of the requirement of FoxN3 in chondrocranium development. In addition, we analyse the effects of FoxN3 knockdown on cranial muscle development. We show that FoxN3 knockdown primarily affects the novel skeletal structures unique to anuran larvae, i.e. the rostralia or the fine structure of the gill apparatus. The articulation between the infrarostral and Meckel's cartilage is malformed and the filigreed processes of the gill basket do not develop. Because these features do not develop after FoxN3 knockdown, the head morphology resembles that in the less specialised larvae of salamanders. Furthermore, the development of all cartilages derived from the neural crest is delayed and cranial muscle fibre development incomplete. The cartilage precursors initially condense in their proper position but later differentiate incompletely; several visceral arch muscles start to differentiate at their origin but fail to extend toward their insertion. Our findings indicate that FoxN3 is essential for the development of novel cartilages such as the infrarostral and other cranial tissues derived from the neural crest and, indirectly, also for muscle morphogenesis.


Too much of too little: xylitol, an unusual trigger of a chronic metabolic hyperchloremic acidosis.
Homeopathic globules are frequently used in children as a first-line treatment. Most of these globules are coated with sugar substitutes like xylitol; these substitutes are known for their laxative effect. Our patient shows that consumption of globules coated with xylitol does not have only laxative effects. It may cause indeed considerable weight loss and life-threatening enteral bicarbonate loss by diarrhea when overdosed in an infant.


Characterization of T-cell non-Hodgkin's lymphoma and its association with Epstein-Barr virus in Pakistani patients.
This study analyzes the prevalence of T-cell non-Hodgkin's lymphoma (T-NHL) in a major referral center of Pakistan and its association with Epstein-Barr virus (EBV). Ninety-two cases of T-NHL were characterized on the basis of morphology, immunohistochemistry and genetic features. The prevalence of T-NHL was 22.2% of the total NHLs diagnosed during the eight years period (1992-1999). Polymerase chain reaction (PCR) technique was used to assess T-cell clonality in paraffin-embedded tissues of known T-NHL. Amplifiable DNA was isolated from all the cases, which were further studied for T-cell receptor (TcR)-beta, gamma, delta, and IgH chain gene rearrangements. Out of 92 cases 51 cases showed clonal product and 33 demonstrated polyclonal smear for beta, gamma, or delta chain genes, respectively, whereas 8 cases exhibited IgH chain gene rearrangement for FR2 region. This study demonstrated frequent presence of EBV in T-NHL (55.4%) by PCR, which were further tested for the localization of the virus by in situ hybridization (ISH). The extent of polymorphism in EBV genome was studied by single stranded conformation polymorphism (SSCP) technique for Bam HI E, K, N and Z regions. Hypervariability in Bam HI K, and N regions was noticeably higher compared to E or Z regions. In conclusion, our study demonstrated that the prevalence of T-NHL in Pakistan is slightly higher to that reported for Western communities. In addition, the frequency of EBV genome in T-NHL is intermediate as compared to other studies. No association was established between EBV variants differentiated on the basis of sequence heterogeneity in Bam HI K, N, E and Z regions with the manifestation of different subsets of T-NHL.


Phenomenological-narrative contributions to understand the narrative strategies that shape the autobiographical account throughout different moments of the therapeutic process.
Different studies suggest that the strategies and narrative styles that people use to construct their autobiographical accounts have repercussions on their self-organization, as well as on their identity experience and their conception of the world. Empirical evidence supports changes in different aspects related to process, structure, and content in the narrative of clients during the course of the therapeutic process; these, in turn, seem to condition the course and the results of the process. In this paper we will seek to show, based on a case study and through the application of a method of phenomenological-narrative analysis, what are the predominant narrative strategies that a client uses in order to shape her autobiographical narrative in the initial and final moments of her psychotherapeutic process. Our data suggest that the narrative strategies at the beginning and end of the therapy are qualitatively differentiable. Changes are observed in the plot of the respective accounts, as well as a differentiated mode in the use of narrative functions. More specifically toward the end of psychotherapy, the client makes a deeper characterization of herself and others, based on predicates of a subjectivating, interpretive, and evaluative-reflective kind. She also predominantly uses proconcluding metacomments, which could facilitate the integration of problematic experiences. In summary, our data suggest that after a successful therapeutic process the client uses more complex and integrated narrative strategies for the construction of her autobiographical account.


NMR Chemical Shift Mapping of SH2 Peptide Interactions.
Heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) experiments offer a rapid and high resolution approach to gaining binding and conformational insights into a protein-peptide interaction. By tracking 1H and 15N chemical shift changes over the course of a peptide titration into isotopically labeled protein, amide NH pairs of amino acids whose chemical environment changes upon peptide binding can be identified. When mapped onto a structure of the protein, this approach can identify the peptide-binding interface or regions undergoing conformation changes within a protein upon ligand binding. Monitoring NMR chemical shift changes can also serve as a screening technique to identify novel interaction partners for a protein or to determine the binding affinity of a weak protein-peptide interaction. Here, we describe the application of NMR chemical shift mapping to the study of peptide binding to the C-terminal SH2 domain of PLCγ1.


Passive and Active Social Media Use and Depressive Symptoms Among United States Adults.
Social media allows users to explore self-identity and express emotions or thoughts. Research looking into the association between social media use (SMU) and mental health outcomes, such as anxiety or depressive symptoms, have produced mixed findings. These contradictions may best be addressed by examining different patterns of SMU as they relate to depressive symptomatology. We sought to assess the independent associations between active versus passive SMU and depressive symptoms. For this, we conducted an online survey of adults 18-49 of age. Depressive symptoms were measured using the Patient-Reported Outcomes Measurement Information System brief depression scale. We measured active and passive SMU with previously developed items. Factor analysis was used to explore the underlying factor structure. Then, we used ordered logistic regression to assess associations between both passive and active SMU and depressive symptoms while controlling for sociodemographic covariates. Complete data were received from 702 participants. Active and passive SMU items loaded on separate factors. In multivariable analyses that controlled for all covariates, each one-point increase in passive SMU was associated with a 33 percent increase in depressive symptoms (adjusted odds ratio [AOR] = 1.33, 95 percent confidence interval [CI] = 1.17-1.51). However, in the same multivariable model, each one-point increase in active SMU was associated with a 15 percent decrease in depressive symptoms (AOR = 0.85, 95 percent CI = 0.75-0.96). To inform interventions, future research should determine directionality of these associations and investigate related factors.


Gremlin 2 promotes differentiation of embryonic stem cells to atrial fate by activation of the JNK signaling pathway.
The bone morphogenetic protein antagonist Gremlin 2 (Grem2) is required for atrial differentiation and establishment of cardiac rhythm during embryonic development. A human Grem2 variant has been associated with familial atrial fibrillation, suggesting that abnormal Grem2 activity causes arrhythmias. However, it is not known how Grem2 integrates into signaling pathways to direct atrial cardiomyocyte differentiation. Here, we demonstrate that Grem2 expression is induced concurrently with the emergence of cardiovascular progenitor cells during differentiation of mouse embryonic stem cells (ESCs). Grem2 exposure enhances the cardiogenic potential of ESCs by 20-120-fold, preferentially inducing genes expressed in atrial myocytes such as Myl7, Nppa, and Sarcolipin. We show that Grem2 acts upstream to upregulate proatrial transcription factors CoupTFII and Hey1 and downregulate atrial fate repressors Irx4 and Hey2. The molecular phenotype of Grem2-induced atrial cardiomyocytes was further supported by induction of ion channels encoded by Kcnj3, Kcnj5, and Cacna1d genes and establishment of atrial-like action potentials shown by electrophysiological recordings. We show that promotion of atrial-like cardiomyocytes is specific to the Gremlin subfamily of BMP antagonists. Grem2 proatrial differentiation activity is conveyed by noncanonical BMP signaling through phosphorylation of JNK and can be reversed by specific JNK inhibitors, but not by dorsomorphin, an inhibitor of canonical BMP signaling. Taken together, our data provide novel mechanistic insights into atrial cardiomyocyte differentiation from pluripotent stem cells and will assist the development of future approaches to study and treat arrhythmias.


Zinc Nutrition and Inflammation in the Aging Retina.
Zinc is an essential nutrient for human health. It plays key roles in maintaining protein structure and stability, serves as catalytic factor for many enzymes, and regulates diverse fundamental cellular processes. Zinc is important in affecting signal transduction and, in particular, in the development and integrity of the immune system, where it affects both innate and adaptive immune responses. The eye, especially the retina-choroid complex, has an unusually high concentration of zinc compared to other tissues. The highest amount of zinc is concentrated in the retinal pigment epithelium (RPE) (RPE-choroid, 292 ± 98.5 µg g-1 dry tissue), followed by the retina (123 ± 62.2 µg g-1 dry tissue). The interplay between zinc and inflammation has been explored in other parts of the body but, so far, has not been extensively researched in the eye. Several lines of evidence suggest that ocular zinc concentration decreases with age, especially in the context of age-related disease. Thus, a hypothesis that retinal function could be modulated by zinc nutrition is proposed, and subsequently trialled clinically. In this review, the distribution and the potential role of zinc in the retina-choroid complex is outlined, especially in relation to inflammation and immunity, and the clinical studies to date are summarized.


Functional and Mechanistic Neurotoxicity Profiling Using Human iPSC-Derived Neural 3D Cultures.
Neurological disorders affect millions of people worldwide and appear to be on the rise. Whereas the reason for this increase remains unknown, environmental factors are a suspected contributor. Hence, there is an urgent need to develop more complex, biologically relevant, and predictive in vitro assays to screen larger sets of compounds with the potential for neurotoxicity. Here, we employed a human induced pluripotent stem cell (iPSC)-based 3D neural platform composed of mature cortical neurons and astrocytes as a model for this purpose. The iPSC-derived human 3D cortical neuron/astrocyte co-cultures (3D neural cultures) present spontaneous synchronized, readily detectable calcium oscillations. This advanced neural platform was optimized for high-throughput screening in 384-well plates and displays highly consistent, functional performance across different wells and plates. Characterization of oscillation profiles in 3D neural cultures was performed through multi-parametric analysis that included the calcium oscillation rate and peak width, amplitude, and waveform irregularities. Cellular and mitochondrial toxicity were assessed by high-content imaging. For assay characterization, we used a set of neuromodulators with known mechanisms of action. We then explored the neurotoxic profile of a library of 87 compounds that included pharmaceutical drugs, pesticides, flame retardants, and other chemicals. Our results demonstrated that 57% of the tested compounds exhibited effects in the assay. The compounds were then ranked according to their effective concentrations based on in vitro activity. Our results show that a human iPSC-derived 3D neural culture assay platform is a promising biologically relevant tool to assess the neurotoxic potential of drugs and environmental toxicants.


Assembling the Career Firefighter Health Study cohort: A methods overview.
Studies of World Trade Center (WTC)-exposed rescue/recovery workers report the increased occurrence of health conditions after work at the WTC disaster site. However, the extent to which these associations are due to WTC exposure is unclear, in part due to the lack of suitable comparison groups. Accordingly, we identified a previously assembled National Institute for Occupational Safety and Health (NIOSH) cohort of career firefighters from three US cities (n = 29,992). Here, we document the challenges in establishing this non-WTC-exposed firefighter cohort for the goal of tracking and comparing cancer and chronic health conditions in WTC-exposed and non-WTC-exposed firefighters. Follow-up process included institutional review board applications, data use agreements, state cancer registry linkages and vital status determination for the NIOSH firefighter cohort. After completion of these steps, we undertook outreach to the three original city fire departments and union officials, before contact tracing and direct recruitment of 14,566 living firefighters to complete a confidential health survey. We staggered recruitment efforts by the city, using letters, postcards, emails, videos, and telephone outreach. Participants who completed the survey received $10. A total of 4962 of 14,566 alive firefighters responded to the baseline survey (34.1% response rate). Respondents were older and more likely to be non-Hispanic white than nonrespondents. We provide an overview of the process for the first survey to collect information on physical and mental health conditions among US firefighters. The data collected will have an important impact on studies of WTC rescue/recovery work, firefighting, and related health conditions.


Kernel and regression spline smoothing techniques to estimate coefficient in rates model and its application in psoriasis.
Background: Data types are recurrent events in studies in which each person may experience an event at different times. One of the most popular approaches to analyze recurrent event data is obtaining an estimate of the means/rate of events at different times. In this context, determining the variability over time can help better understand the effect of factor on the response. In this study, we applied smoothing methods to estimate coefficients in time-dependent rate model, and we also showed its application in data of psoriasis patients. Methods: In the present study, psoriasis patients who experienced relapse that led to hospitalization during 2005 and 2014 in the Dermatology Department of Imam Khomeini hospital in Tehran were examined. To investigate the rate of relapse during a year, time-dependent rate model was used and variability of the effects was assessed using Wald test. Both b-spline and kernel methods were used to estimate time varying coefficients in rates model. Finally, results from methods were compared based on the obtained estimates. Results: Based on the results of the Wald test, the effect of season on the occurrence of psoriasis was significantly different (p<0.01). Also according to the estimated coefficients from both b-spline and kernel methods, there was little difference between them. Conclusion: In situations in which the effect of a variable is different at different times, using time-dependent coefficients rate model can lead to a better estimate of the effect of variable on the response. On the other hand, smoothing methods can smooth the effects of the variables that vary over time.


E-cadherin binds to desmoglein to facilitate desmosome assembly.
Desmosomes are adhesive junctions composed of two desmosomal cadherins: desmocollin (Dsc) and desmoglein (Dsg). Previous studies demonstrate that E-cadherin (Ecad), an adhesive protein that interacts in both trans (between opposing cells) and cis (on the same cell surface) conformations, facilitates desmosome assembly via an unknown mechanism. Here we use structure-function analysis to resolve the mechanistic roles of Ecad in desmosome formation. Using AFM force measurements, we demonstrate that Ecad interacts with isoform 2 of Dsg via a conserved Leu-175 on the Ecad cis binding interface. Super-resolution imaging reveals that Ecad is enriched in nascent desmosomes, supporting a role for Ecad in early desmosome assembly. Finally, confocal imaging demonstrates that desmosome assembly is initiated at sites of Ecad mediated adhesion, and that Ecad-L175 is required for efficient Dsg2 and desmoplakin recruitment to intercellular contacts. We propose that Ecad trans interactions at nascent cell-cell contacts initiate the recruitment of Dsg through direct cis interactions with Ecad which facilitates desmosome assembly.