Analysis of regional scale risk of whirling disease in populations of Colorado and Rio Grande cutthroat trout using a Bayesian belief network model.
Introduction and spread of the parasite Myxobolus cerebralis, the causative agent of whirling disease, has contributed to the collapse of wild trout populations throughout the intermountain west. Of concern is the risk the disease may have on conservation and recovery of native cutthroat trout. We employed a Bayesian belief network to assess probability of whirling disease in Colorado River and Rio Grande cutthroat trout (Oncorhynchus clarkii pleuriticus and Oncorhynchus clarkii virginalis, respectively) within their current ranges in the southwest United States. Available habitat (as defined by gradient and elevation) for intermediate oligochaete worm host, Tubifex tubifex, exerted the greatest influence on the likelihood of infection, yet prevalence of stream barriers also affected the risk outcome. Management areas that had the highest likelihood of infected Colorado River cutthroat trout were in the eastern portion of their range, although the probability of infection was highest for populations in the southern, San Juan subbasin. Rio Grande cutthroat trout had a relatively low likelihood of infection, with populations in the southernmost Pecos management area predicted to be at greatest risk. The Bayesian risk assessment model predicted the likelihood of whirling disease infection from its principal transmission vector, fish movement, and suggested that barriers may be effective in reducing risk of exposure to native trout populations. Data gaps, especially with regard to location of spawning, highlighted the importance in developing monitoring plans that support future risk assessments and adaptive management for subspecies of cutthroat trout.


Screening transthyretin amyloid fibril inhibitors: characterization of novel multiprotein, multiligand complexes by mass spectrometry.
Tetrameric transthyretin is involved in transport of thyroxine and, through its interactions with retinol binding protein, vitamin A. Dissociation of these structures is widely accepted as the first step in the formation of transthyretin amyloid fibrils. Using a mass spectrometric approach, we have examined a series of 18 ligands proposed as inhibitors of this process. The ligands were evaluated for their ability to bind to and stabilize the tetrameric structure, their cooperativity in binding, and their ability to compete with the natural ligand thyroxine. The observation of a novel ten-component complex containing six protein subunits, two vitamin molecules, and two synthetic ligands allows us to conclude that ligand binding does not inhibit association of transthyretin with holo retinol binding protein.


14-3-3 binds to and mediates phosphorylation of microtubule-associated tau protein by Ser9-phosphorylated glycogen synthase kinase 3beta in the brain.
In mammalian brain, tau, glycogen synthase kinase 3beta (GSK3beta), and 14-3-3, a phosphoserine-binding protein, are parts of a multiprotein tau phosphorylation complex. Within the complex, 14-3-3 simultaneously binds to tau and GSK3beta (Agarwal-Mawal, A., Qureshi, H. Y., Cafferty, P. W., Yuan, Z., Han, D., Lin, R., and Paudel, H. K. (2003) J. Biol. Chem. 278, 12722-12728). The molecular mechanism by which 14-3-3 connects GSK3beta to tau within the complex is not clear. In this study, we find that GSK3beta within the tau phosphorylation complex is phosphorylated on Ser(9). From extracts of rat brain and rat primary cultured neurons, Ser(9)-phosphorylated GSK3beta precipitates with glutathione-agarose beads coated with glutathione S-transferase-14-3-3. Similarly, from rat brain extract, Ser(9)-phosphorylated GSK3beta co-immunoprecipitates with tau. In vitro, 14-3-3 binds to GSK3beta only when the kinase is phosphorylated on Ser(9). In transfected HEK-293 cells, 14-3-3 binds to Ser(9)-phosphorylated GSK3beta and does not bind to GSK3beta (S9A). Tau, on the other hand, binds to both GSK3beta (WT) and GSK3beta (S9A). Moreover, 14-3-3 enhances the binding of tau with Ser(9)-phosphorylated GSK3beta by approximately 3-fold but not with GSK3beta (S9A). Similarly, 14-3-3 stimulates phosphorylation of tau by Ser(9)-phosphorylated GSK3beta but not by GSK3beta (S9A). In transfected HEK-293 cells, Ser(9) phosphorylation suppresses GSK3beta-catalyzed tau phosphorylation in the absence of 14-3-3. In the presence of 14-3-3, however, Ser(9)-phosphorylated GSK3beta remains active and phosphorylates tau. Our data indicate that within the tau phosphorylation complex, 14-3-3 connects Ser(9)-phosphorylated GSK3beta to tau and Ser(9)-phosphorylated GSK3beta phosphorylates tau.


Clinicopathologic implications of the BRAF(V600E) mutation in papillary thyroid cancer: a subgroup analysis of 3130 cases in a single center.
The BRAF mutation has been shown to be associated with aggressive clinicopathologic characteristics of papillary thyroid cancer (PTC). However, several studies that analyzed hundreds of patients have not demonstrated any correlation. The objective of this study was to investigate the relationship of the BRAF mutation with clinicopathologic factors in a large group of homogenous PTC patients. We collected data of PTC patients who received curative resection of the thyroid gland and who had undergone BRAF mutation tests of their thyroid cancer tissue. Minor variant PTCs and mixed-type thyroid cancers were excluded in this analysis. Clinicopathologic characteristics, including age, sex, BRAF mutation, tumor histology, size, extrathyroidal extension, tumor margin, lymph node metastasis, multifocality, stage, and associated thyroid disease, were collected. The relationship of the BRAF mutation with clinicopathologic factors was analyzed in each homogenous histologic PTC. There were 3130 PTC patients who met the criteria, and these patients were divided into three major histologic groups: conventional PTC (n = 2947), diffuse sclerosing variant PTC (n = 98), and follicular variant PTC (n = 85). The BRAF mutation was variably detected in 75.3%, 61%, and 40% of patients, respectively. In conventional PTC cases, the BRAF mutation was significantly associated with large tumor size, extrathyroidal extension, and lymph node metastasis. Coexistent chronic lymphocytic thyroiditis was significantly less prevalent in the BRAF mutant group. Age, sex, and tumor margin status were not significantly correlated with the BRAF status. There was no evidence that any clinicopathologic factors were linked with the BRAF mutation status in diffuse sclerosing and follicular variant PTCs. The BRAF mutation was differentially detected in each histologic subtype of PTC and was strongly correlated with pathologic factors, most strongly with no coexistent chronic lymphocytic thyroiditis, in conventional PTC. The BRAF mutation is suggested to be a poor prognostic marker in conventional PTC, and the BRAF mutational analysis may lead to better management for individual PTC patients.


Identifying and addressing barriers to African American and non-African American families' discussions about preemptive living related kidney transplantation.
Ethnic/racial minority and nonminority families' perceived barriers to discussing preemptive living related kidney transplantation (LRKT) and their views on the potential value of health care professionals trained to address barriers are unknown. OBJECTIVE, SETTING, AND PARTICIPANTS: To collect pilot data for evaluating perceived barriers to preemptive LRKT and to inform the development of a culturally sensitive intervention to improve families' consideration of LRKT. In 4 structured group interviews of African American and non-African American patients (2 groups) with progressing chronic kidney disease and their family members (2 groups), participants' perceived barriers to initiating LRKT discussions and their views regarding the value of social workers to support discussions were explored. Patients' barriers included concerns about their (1) ability to initiate discussions, (2) discussions being misinterpreted as donation requests, (3) potential burdening of family members, (4) uncertainty about when to initiate discussions, and (5) inducing guilt or coercing family members. Family members' barriers included (1) feeling overwhelmed by patients' illness, (2) patients' denial about their illness, (3) caregiver stress, and (4) uncertainty about their own health or the health of other family members who might donate or need a kidney in the future. Participants reported that social workers could facilitate difficult or awkward discussions and help families understand the LRKT process, address financial concerns, and cope emotionally. Themes were similar between African Americans and non-African Americans. Families identified several barriers to discussing preemptive LRKT that could be addressed by social workers. Further research must be done to determine whether social workers need to tailor interventions to address families' cultural differences.


Luminex LiquiChip System for the Evaluation of Cytokine Levels in Patients with Contact Dermatitis to Potassium Dichromate.
Allergic contact dermatitis (ACD) is associated with increased production of cytokines. The patch test is the "gold-standard" diagnostic method, but it poses a risk of false results. To evaluate a novel laboratory technique, the Luminex LiquiChip, which simultaneously measures blood levels of multiple cytokines, as a diagnostic tool in patients with chrome-induced ACD. The study group included 20 patients with ACD and relevant patch test results for potassium dichromate and 19 patients with ACD for nickel or fragrance as control. Peripheral blood mononuclear cells (PBMCs) were cultured in the presence and absence of potassium dichromate. The Luminex LiquiChip was used to measure levels of the following cytokines: granulocyte-macrophage colony-stimulating factor, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, interferon-γ, and tumor necrosis factor (TNF)-α. Potassium dichromate-stimulated PBMCs secreted significantly higher amounts of all cytokines except TNF-α than nonstimulated PBMCs. PBMCs from patients with ACD to chromium secreted significantly higher amounts of all cytokines tested, except IL-4, compared to PBMCs from patients with ACD to nickel or fragrance. Potassium dichromate stimulates the production of both Th1- and Th2-type cytokines in patients with chrome allergy. The Luminex LiquiChip is a promising in vitro method and may serve as a diagnostic tool for ACD.


Luminescence screening assays for the identification of sensitizers for lanthanides based on the controlled formation of ternary lanthanide complexes with DTPA-bisamide ligands.
The formation of ternary complexes of lanthanide-diethylenetriamine pentaacetic acid (DTPA)-bisamide complexes with different aromatic acids and their application in luminescent screening assays are presented. The europium complexes of DTPA-bisethylamide (BEA), DTPA-bisbutylamide (BUA), DTPA-bis(2-norbornyl) (NBA), and DTPA-bis(1-adamantyl) (ADA) have been isolated and the sensitization of emission upon 1:1 formation with aromatic acids has been studied by luminescence spectroscopy. The ternary complexes show stronger luminescence with picolinate (PCA) rather than phthalate (PTA) or benzoate (BZA), with the latter forming 1:2 complexes. Isophthalate and dipicolinate sensitizers do not show a 1:1 formation of the ternary complexes. Electrospray mass spectra show characteristic peaks that confirm the formation of the ternary complexes, and NMR spectroscopic studies demonstrated a conformational locking effect upon formation of the ternary complex. It is shown that europium complexes with bulkier amide arms provide complexes with stronger luminescence enhancement that is clearly attributable to the bulkiness of the amide arms. In a luminescence screening experiment, different substitutions on BZA and PCA were examined. Sensitizers with a long alkyl chain show greater luminescent enhancements than the nonsubstituted acids, which can be attributed to an "umbrella" effect of the alkyl chain that protects the europium luminescent center from the quenching of secondary high-energy vibrations, in particular O-H from water molecules. The same effect is presented for the quinoline derivatives as sensitizers. Quinoline derivatives with salicylate binding units were identified as the best sensitizers when combined with EuADA, which is even bulkier than EuNBA, with HQ-3COO showing a 50-fold enhancement of emission upon formation of 1:1 complexes.


From structure to function in the lateralized brain: how structural properties of the arcuate and uncinate fasciculus are associated with dichotic listening performance.
Structural asymmetries in white matter tracts within the language system have been suggested to be one of the factors underlying functional language lateralization. To test this assumption, the present study examined how performance in the dichotic listening task, a behavioral measure of language dominance, is affected by macro- and microstructural properties of the arcuate and uncinate fasciculus. To this end, whole brain tractography was performed on 29 diffusion tensor imaging datasets obtained from healthy adult participants. Mean tract volume and fractional anisotropy of the uncinate and arcuate fasciculus were linked to the individual extent of the right ear advantage in the dichotic listening task. On the macrostructural level, both arcuate and uncinate fasciculus had a larger tract volume in the left compared to the right hemisphere. In contrast, fractional anisotropy was higher in the right than in the left arcuate fasciculus. These structural asymmetries were linked to functional lateralization, that is, tract volume and fractional anisotropy of the left arcuate fasciculus were positively correlated to the strength of functional language lateralization, as was the volume of the right uncinate fasciculus. In conclusion, the results of the present study suggest that both micro- and macro-structural properties of language-relevant intrahemispheric white matter tracts modulate the behavioral correlates of language lateralization.


Role of renal vascular potassium channels in physiology and pathophysiology.
The control of renal vascular tone is important for the regulation of salt and water balance, blood pressure and the protection against damaging elevated glomerular pressure. The K+ conductance is a major factor in the regulation of the membrane potential (Vm ) in vascular smooth muscle (VSMC) and endothelial cells (EC). The vascular tone is controlled by Vm via its effect on the opening probability of voltage-operated Ca2+ channels (VOCC) in VSMC. When K+ conductance increases Vm becomes more negative and vasodilation follows, while deactivation of K+ channels leads to depolarization and vasoconstriction. K+ channels in EC indirectly participate in the control of vascular tone by endothelium-derived vasodilation. Therefore, by regulating the tone of renal resistance vessels, K+ channels have a potential role in the control of fluid homoeostasis and blood pressure as well as in the protection of the renal parenchyma. The main classes of K+ channels (calcium activated (KCa ), inward rectifier (Kir ), voltage activated (Kv ) and ATP sensitive (KATP )) have been found in the renal vessels. In this review, we summarize results available in the literature and our own studies in the field. We compare the ambiguous in vitro and in vivo results. We discuss the role of single types of K+ channels and the integrated function of several classes. We also deal with the possible role of renal vascular K+ channels in the pathophysiology of hypertension, diabetes mellitus and sepsis.


NanoCrystal: A Web-Based Crystallographic Tool for the Construction of Nanoparticles Based on Their Crystal Habit.
Modeling of nanoparticles is an essential first step to assess their capacities for different uses such as energy storage and drug delivery. However, creating an initial starting conformation for modeling and simulation is tedious because every crystalline material grows with a different crystal habit. In this application note, we describe NanoCrystal, a novel web-based crystallographic tool that creates nanoparticle models from any crystal structure guided by their preferred equilibrium shape under standard conditions according to the Wulff morphology (crystal habit). Users can upload a cif file, define the Miller indices and their corresponding minimum surface energies according to the Wulff construction of a particular crystal, and specify the size of the nanocrystal. As a result, the nanoparticle is constructed and visualized, and the coordinates of the atoms are output to the user. NanoCrystal can be accessed at http://nanocrystal.vi-seem.edu/ .


Metabolic syndrome in HIV-infected patients from an urban, midwestern US outpatient population.
The association between the use of highly active antiretroviral therapy (HAART) and an increased risk of metabolic syndrome and cardiovascular disease remains unclear. We conducted a prospective, cross-sectional study of the risk factors associated with metabolic syndrome and cardiovascular disease among patients from an urban outpatient human immunodeficiency virus (HIV) clinic. Evaluation included laboratory data that were obtained after an overnight fast and a health survey that assessed traditional risk factors associated with cardiovascular disease, HIV-related factors, and comorbidities. Data collected were compared with data files from a cohort from the National Health and Nutrition Examination Survey (NHANES; 2001-2002) of persons who were seronegative for HIV infection who were matched for age, sex, race, and tobacco use. Four hundred seventy-one HIV-infected subjects provided complete data. The overall prevalence of metabolic syndrome was similar between the group HIV-infected patients and the group of persons who were seronegative for HIV infection (25.5% vs. 26.5%, respectively), although the HIV-infected patients had a significantly smaller waist circumference, lower body mass index, lower high-density lipoprotein cholesterol levels, higher triglyceride levels, and lower glucose levels, compared with the subjects from the NHANES cohort. Framingham 10-year risk scores were also similar between the 2 groups. HIV-infected patients with metabolic syndrome were more likely to be diabetic, older, and white and have a high CD4 cell count and body mass index, compared with patients without metabolic syndrome (P<.05 for all). The type or duration of antiretroviral therapy was not an independent risk factor for metabolic syndrome. The prevalence of metabolic syndrome is high among HIV-infected persons, but not higher than the prevalence among HIV-uninfected persons. Traditional risk factors play a more significant role in the development of metabolic syndrome than do HIV treatment-associated factors.


An online operational rainfall-monitoring resource for epidemic malaria early warning systems in Africa.
Periodic epidemics of malaria are a major public health problem for many sub-Saharan African countries. Populations in epidemic prone areas have a poorly developed immunity to malaria and the disease remains life threatening to all age groups. The impact of epidemics could be minimized by prediction and improved prevention through timely vector control and deployment of appropriate drugs. Malaria Early Warning Systems are advocated as a means of improving the opportunity for preparedness and timely response. Rainfall is one of the major factors triggering epidemics in warm semi-arid and desert-fringe areas. Explosive epidemics often occur in these regions after excessive rains and, where these follow periods of drought and poor food security, can be especially severe. Consequently, rainfall monitoring forms one of the essential elements for the development of integrated Malaria Early Warning Systems for sub-Saharan Africa, as outlined by the World Health Organization. The Roll Back Malaria Technical Resource Network on Prevention and Control of Epidemics recommended that a simple indicator of changes in epidemic risk in regions of marginal transmission, consisting primarily of rainfall anomaly maps, could provide immediate benefit to early warning efforts. In response to these recommendations, the Famine Early Warning Systems Network produced maps that combine information about dekadal rainfall anomalies, and epidemic malaria risk, available via their Africa Data Dissemination Service. These maps were later made available in a format that is directly compatible with HealthMapper, the mapping and surveillance software developed by the WHO's Communicable Disease Surveillance and Response Department. A new monitoring interface has recently been developed at the International Research Institute for Climate Prediction (IRI) that enables the user to gain a more contextual perspective of the current rainfall estimates by comparing them to previous seasons and climatological averages. These resources are available at no cost to the user and are updated on a routine basis.


Characterisation of mental health conditions in social media using Informed Deep Learning.
The number of people affected by mental illness is on the increase and with it the burden on health and social care use, as well as the loss of both productivity and quality-adjusted life-years. Natural language processing of electronic health records is increasingly used to study mental health conditions and risk behaviours on a large scale. However, narrative notes written by clinicians do not capture first-hand the patients' own experiences, and only record cross-sectional, professional impressions at the point of care. Social media platforms have become a source of 'in the moment' daily exchange, with topics including well-being and mental health. In this study, we analysed posts from the social media platform Reddit and developed classifiers to recognise and classify posts related to mental illness according to 11 disorder themes. Using a neural network and deep learning approach, we could automatically recognise mental illness-related posts in our balenced dataset with an accuracy of 91.08% and select the correct theme with a weighted average accuracy of 71.37%. We believe that these results are a first step in developing methods to characterise large amounts of user-generated content that could support content curation and targeted interventions.


Children are not little adults: just ask their hematopoietic stem cells.
HSCs differ during ontogeny in some important parameters, including anatomic site of residence and cell cycling characteristics. In this issue of the JCI, Bowie et al. show that postnatal HSCs as well as fetal liver HSCs in mice are active in the cell cycle at much higher rates than that of adult HSCs; however, this increased frequency of cycling abruptly ceases 4 weeks after birth (see the related article beginning on page 2808). The cycling postnatal HSCs expressed high levels of CXC chemokine ligand 12 (CXCL12, also known as stromal cell-derived factor 1 [SDF-1]), a chemokine previously implicated in stem cell trafficking to the marrow cavity and shown to be expressed by cells within the hematopoietic microenvironment. These cells also possessed an engraftment defect impeding reconstitution in irradiated recipient mice, which was reversible by pretransplant administration of antagonists of the CXCL12 receptor, CXCR4. Such agents are currently clinically available, suggesting that this approach could be used to improve stem cell transplantation and engraftment.


Improving peer engagement of children with autism on the school playground: a randomized controlled trial.
This study aimed to test the effects of a psychosocial intervention, Remaking Recess, on peer engagement for children with autism spectrum disorder (ASD). Using a randomized, wait-list-controlled design, the intervention was implemented during recess at four elementary schools. The immediate treatment (IT) group consisted of 13 (2 female) elementary school students with ASD and the wait-list (WL) group contained 11 (4 female) students with ASD. All of the children with ASD were fully included in the general education program. Analyses revealed that time spent engaged with peers was significantly increased for the IT group and maintained over the follow-up. School playground staff in the IT group showed increased behaviors aimed at improving peer engagement for children with ASD compared to playground staff at the WL sites. These improvements did not maintain to follow-up. These results suggest that a low dose, brief intervention can be beneficial in increasing peer engagement for children with autism in inclusive settings, but continued support of playground staff is likely needed.


Aripiprazole versus placebo for schizophrenia.
First generation 'typical' antipsychotics such as chlorpromazine and haloperidol have been the mainstay of treatment up until the introduction of the second generation 'atypical' antipsychotics such as risperidone and olanzapine. Typical and atypical antipsychotics do provide a treatment response for most people with schizophrenia, whether a reduction in psychotic episodes or a lessening in the severity of their illness. However, a proportion of people still do not respond adequately to antipsychotic medication. Additionally, atypical and especially typical antipsychotics are associated with serious adverse effects, which can often compromise compliance with medication and therefore increase the incidences of relapse. In this review we examine the effects of aripiprazole compared with placebo. To evaluate the effects of aripiprazole compared with placebo for people with schizophrenia and schizophrenia-like psychoses. We searched the Cochrane Schizophrenia Group Trials Register (January 2008) which is based on regular searches of BIOSIS, CENTRAL, CINAHL, EMBASE, MEDLINE and PsycINFO. For this update, we carried out an initial search in May 2007 and a second search in August 2008. We included all randomised trials comparing aripiprazole with placebo in people with schizophrenia or schizophrenia-like psychosis. We extracted data independently. For dichotomous data we calculated risk ratios (RR) and their 95% confidence intervals (CI) on an intention-to-treat basis based on a fixed-effect model. We calculated numbers needed to treat/harm (NNT/NNH) where appropriate. For continuous data, we calculated mean differences (MD) again based on a fixed-effect model. Despite the fact that 2585 people participated in nine randomised aripiprazole studies, we were unable to extract any usable data on death, service outcomes, general functioning, behaviour, engagement with services, satisfaction with treatment; economic outcomes or cognitive functioning. In general, study attrition was very large for all studies over four weeks' duration. There was high attrition in most of the included studies. Fewer people left the aripiprazole group compared with those in the placebo group (n = 2585, 9 RCTs, RR 0.73 CI 0.60 to 0.87). Compared with placebo, aripiprazole significantly decreased relapse in both the short (n = 310, 1 RCT, RR 0.59 CI 0.45 to 0.77) and medium term (n = 310, 1 RCT, RR 0.66 CI 0.53 to 0.81). It also produced better compliance with study protocol (n = 2275, 8 RCTs, RR 0.74 CI 0.59 to 0.93). Aripiprazole may decrease prolactin levels below those expected from placebo (n = 305, 2 RCT, RR 0.21 CI 0.11 to 0.37). Insomnia (˜23%) and headaches (˜15%) were commonly reported in both groups, with no significant difference. Aripiprazole may be effective for the treatment of schizophrenia. Aripiprazole has a lower risk of raised prolactin and prolongation of the QTc interval. Clearly reported pragmatic short-, medium- and long-term randomised controlled trials should be undertaken to determine its position in everyday clinical practice.


Glioma tropism of lentivirally transduced hematopoietic progenitor cells.
Experimental gliomas attract hematopoietic progenitor cells (HPC) in vivo. HPC are therefore promising candidates for a cell-based delivery of therapeutic molecules to experimental gliomas. A therapeutic application requires efficient genetic manipulation of the cellular vector and a lack of tumorigenicity. Here, we studied the impact of lenti-viral transduction on the glioma tropism of human or murine HPC. Transduction of human or murine HPC with a GFP lentivirus (lenti-GFP) did not interfere with the glioma-mediated attraction of HPC. Bone marrow reconstitution of C57Bl/6 mice with syngeneic GFP-transgenic lineage-depleted bone marrow cells (lin- BM) was as efficient as reconstitution with syngeneic lin- BM transduced ex vivo with lenti-GFP. SMA-560 gliomas growing orthotopically in lenti-GFP-reconstituted VM/Dk mice recruited GFP-positive bone marrow-derived cells. Thus, lentiviral transduction did not interfere with the attraction of exogenously injected HPC or endogenous bone marrow-derived cells by experimental gliomas. Lenti-GFP-HPC implanted directly into tumor-free brains were not tumorigenic. The intravenous injection of lenti-GFP-HPC in glioma-bearing mice did not alter the survival of otherwise untreated animals and had no impact on the survival benefit conferred by cerebral irradiation. Taken together, genetic manipulation of HPC with lenti-GFP neither made these cells tumorigenic nor interfered with their glioma tropism.


Valence Bond Theory Reveals Hidden Delocalized Diradical Character of Polyenes.
The nature of the electronic-structure of polyenes, their delocalization features, and potential diradicaloid characters constitute a fundamental problem in chemistry. To address this problem, we used valence bond self-consistent field (VBSCF) calculations and modeling of polyenes, C2nH2n+2 (n = 2-10). The theoretical treatment shows that starting with n = 5, the polyene's wave function is mainly a shifting 1,4-diradicaloid, a character that increases as the chain length increases, while the contribution of the fundamental Lewis structure with alternating double and single bonds (1) decays quite fast and becomes minor relative to the diradicaloid pack. We show how, nevertheless, it is this wave function that predicts that polyenes will still exhibit alternating short/long CC bonds like the fundamental structure 1. Furthermore, despite the decay of the VB contribution of 1, it remains the single structure with the largest weight among all the individual structures. The mixing of all the 1,4-diradicaloid structures into 1 follows perturbation theory rules, with the result that the delocalization energy due to this mixing is additive and behaves as a linear function of the number of the double bonds, ΔEdel = -6.9 × n (kcal mol-1). The VB modeling shows that while the conjugation stabilizes structure 1, this stabilization energy is energetically overridden by the Pauli repulsion between two adjacent double bonds. Nevertheless, unsubstituted polyenes remain planar; this observation is addressed. Potential manifestations of the diradicaloid nature of polyenes are discussed, and it is concluded that the diradicaloid character is clearly not a well-defined physical property as in real diradicals. Thus, we went full circle to realize that our philosophical question may not be strictly resolved. The localized/delocalized properties of polyenes seem to define a "chemical duality principle". This duality of molecular wave functions is a ubiquitous beguiling phenomenon.


Effects of drinking desalinated seawater on cell viability and proliferation.
Desalination of seawater is becoming an important means to address the increasing scarcity of freshwater resources in the world. Seawater has been used as drinking water in the health, food, and medical fields and various beneficial effects have been suggested, although not confirmed. Given the presence of 63 minerals and trace elements in drinking desalinated seawater (63 DSW), we evaluated their effects on the behavior of tumorigenic and nontumorigenic cells through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and annexin-V-fluorescein isothiocyanate/propidium iodide staining. Our results showed that cell viability and proliferation in the presence of 63 DSW were significantly greater than in mineral water and in the presence of fetal bovine serum in a dose-dependent manner. Furthermore, 63 DSW showed no toxic effect on murine embryonic fibroblast (NIH-3T3) and murine melanoma (B16-F10) cells. In another assay, we also showed that pre-treatment of non-adherent THP-1 cells with 63 DSW reduces apoptosis incidence, suggesting a protective effect against cell death. We conclude that cell viability and proliferation were improved by the mineral components of 63 DSW and this effect can guide further studies on health effects associated with DSW consumption.


Loss of p16 expression is associated with the stem cell characteristics of surface markers and therapeutic resistance in estrogen receptor-negative breast cancer.
Triple-negative breast cancer [TNBC, which is negative for the estrogen receptor (ER), progesterone receptor, and human epidermal growth factor receptor 2] is a high-risk form of the disease without a specific therapy. DNA microarray and immunohistochemical analyses have shown that most TNBCs fall within the basal-like histological subset of breast cancers, which frequently exhibit inactivation of the retinoblastoma tumor suppressor (Rb) and upregulation of the cyclin-dependent kinase inhibitor p16(INK4a) (p16). However, downregulation of p16 expression has been observed in some basal-like breast cancer cell lines, suggesting that such cells can be divided into two groups according to Rb and p16 status. We now show that cells that are CD44(+) and CD24(-) , a phenotype associated with stem-like breast cancer cells, are more abundant in ER(-) /p16(-) breast cancer cell lines than in ER(-) /p16(+) lines. It was also found that p16 expression was downregulated in mammospheres from an ER-negative breast cancer cell line. Depletion of p16 by RNA interference in ER-negative breast cancer cells increased the percentage of CD44(+) /CD24(-) cells and increased the expression of mRNA of the ES-like genes Nanog, Oct4, and Sox2 through an Rb-independent pathway. Furthermore, such depletion of p16 reduced chemosensitivity. The loss of p16 expression may thus reduce the response of ER-negative breast cancer cells to chemotherapy by conferring cancer stem cell-like properties. Consistent with this conclusion, immunohistochemical analysis of the clinical samples suggests that low p16 expression in TNBC is associated with resistance to preoperative chemotherapy.


Clinical application of tumour markers: a review.
Tumour markers have made a difference to oncology practice. They can be used in screening, diagnosis, prognostication and assessment of treatment efficacy. Reports on tumour marker usage suggest that many clinicians assume that a biomarker for a particular cancer can be effectively used for all these indications. This assumption is incorrect. Several guidelines have been published to inform clinicians on effective utilisation of these tests. To outline the recommended uses of the most commonly requested tumours markers in clinical practice. A hand search of literature on the recommended use of carcinoembryonic antigen (CEA), alphafetoprotein (AFP), prostate specific antigen (PSA), CA-125 and CA-19.9. Systematic reviews and prospective randomised clinical trials of tumour marker applications were also looked at. Five key journals and reference lists of relevant studies were considered. Two authors abstracted relevant data independently. Emphasis was given to guidelines from expert panels. The quality of the guidelines was assessed by availability of level of evidence supporting the recommendations. Several national and international expert groups have developed guidelines for use of markers for most cancers. CEA, AFP, PSA, CA-125 and CA-19.9 are validated for use in treatment monitoring of colorectal, hepatocellular, prostatic, ovarian and pancreatic carcinomas respectively. AFP and PSA are also useful for cancer screening in high risk groups. CA-125 has limited role in screening while CEA and CA 19.9 are not recommended for cancer screening. Not all currently available tumour markers can be used for screening and diagnosis of malignancies. Adherence to recommendations on tumour marker utilisation will improve the cost-effectiveness of these tests.


Type VII collagen is a major structural component of anchoring fibrils.
Anchoring fibrils are specialized fibrous structures found in the subbasal lamina underlying epithelia of several external tissues. Based upon their sensitivity to collagenase and the similarity in banding pattern to artificially created segment-long spacing crystallites (SLS) of collagens, several authors have suggested that anchoring fibrils are lateral aggregates of collagenous macromolecules. We recently reported the similarity in length and banding pattern of anchoring fibrils to type VII collagen SLS crystallites. We now report the construction and characterization of a murine monoclonal antibody specific for type VII collagen. The epitope identified by this antibody has been mapped to the carboxyl terminus of the major helical domain of this molecule. The presence of type VII collagen as detected by indirect immunofluorescence in a variety of tissues corresponds exactly with ultrastructural observations of anchoring fibrils. Ultrastructural immunolocalization of type VII collagen using a 5-nm colloidal gold-conjugated second antibody demonstrates metal deposition upon anchoring fibrils at both ends of these structures, as predicted by the location of the epitope on type VII collagen. Type VII collagen is synthesized by primary cultures of amniotic epithelial cells. It is also produced by KB cells (an epidermoid carcinoma cell line) and WISH (a transformed amniotic cell line).


Efficacy of endovascular intervention in patients with unruptured posterior communicating artery aneurysm-related oculomotor nerve palsy.
Oculomotor nerve palsy (ONP) is commonly encountered in daily neurosurgical activities. The ONP secondary to un-ruptured PComA aneurysm might be a unique entity that was different in diagnosis, treatment and prognosis from its ruptured counterparts. Perhaps as a result of the limitation in sample size, studies that solely focused on factors affecting recovery of ONP in patients with unruptured corresponding PComA aneurysms were scarce. In this study, we would like to report a relatively larger case series of patients with un-ruptured PComA aneurysm-related ONP. A retrospective review of medical records of 39 patients with un-ruptured PComA aneurysm-related ONP was performed with endovascular coiling. All 39 consecutive patients underwent endovascular coiling. Eighteen (46%) patients had a complete resolution of ONP, 14 (36%) patients had a partial resolution. Time interval from onset of ONP to endovascular intervention (P=0.004), degree of ONP (P=0.015) and age (P=0.016) were predictors of ONP recovery with statistical significance. Sex, aneurysm size and risk factor exposure (smoking, alcohol abuse and hypertension) were not associated with ONP outcomes. ONP secondary to un-ruptured aneurysm should be treated as a unique entity from its ruptured counterparts. A prospective study that contains surgical clipping and endovascular coiling, and comparison between two treatment modalities would be more convincing and is anticipated.


The characteristic site-specific reactivation phenotypes of HSV-1 and HSV-2 depend upon the latency-associated transcript region.
After replication at sites of initial inoculation, herpes simplex virus type 1 and 2 (HSV-1 and HSV-2) establish lifelong latent infections of the sensory and autonomic neurons of the ganglia serving those sites. Periodically, the virus reactivates from these neurons, and travels centripetally along the neuronal axon to cause recurrent epithelial infection. The major clinically observed difference between infections with herpes simplex virus type 1 and type 2 is the anatomic site specificity of recurrence. HSV-1 reactivates most efficiently and frequently from trigeminal ganglia, causing recurrent ocular and oral-facial lesions, while HSV-2 reactivates primarily from sacral ganglia causing recurrent genital lesions. An intertypic recombinant virus was constructed and evaluated in animal models of recurrent ocular and genital herpes. Substitution of a 2.8-kbp region from the HSV-1 latency-associated transcript (LAT) for native HSV-2 sequences caused HSV-2 to reactivate with an HSV-1 phenotype in both animal models. The HSV-2 phenotype was restored by replacing the mutated sequences with wild-type HSV-2 LAT-region sequences. These sequences or their products must act specifically in the cellular environments of trigeminal and sacral neurons to promote the reactivation patterns characteristic of each virus.


'Poisonous, filthy, loathsome, damnable stuff': the rhetorical ecology of vaccination concern.
In this article, we analyze newspaper articles and advertisements mentioning vaccination from 1915 to 1922 and refer to historical studies of vaccination practices and attitudes in the early 20th century in order to assess historical continuities and discontinuities in vaccination concern. In the Progressive Era period, there were a number of themes or features that resonated with contemporary issues and circumstances: 1) fears of vaccine contamination; 2) distrust of medical professionals; 3) resistance to compulsory vaccination; and 4) the local nature of vaccination concern. Such observations help scholars and practitioners understand vaccine skepticism as longstanding, locally situated, and linked to the sociocultural contexts in which vaccination occurs and is mandated for particular segments of the population. A rhetorical approach offers a way to understand how discourses are engaged and mobilized for particular purposes in historical contexts. Historically situating vaccine hesitancy and addressing its articulation with a particular rhetorical ecology offers scholars and practitioners a robust understanding of vaccination concerns that can, and should, influence current approaches to vaccination skepticism.


Hepatic first-pass uptake of diphenhydramine. A comparative study between fetal and adult sheep.
In this study, two factors that could affect fetal drug exposure were examined: 1) the extent of elimination of drug delivered to the fetal liver from the placenta via the umbilical vein; and 2) the degree to which there is preferential distribution of drug in umbilical venous blood to the fetal upper body, as is the case with oxygen and other endogenous substances. Studies were conducted with the histamine H1 antagonist, diphenhydramine (DPHM), in chronically instrumented nonpregnant and pregnant sheep (115-138 days gestation). Hepatic presystemic DPHM elimination was assessed using simultaneous and separate administration of DPHM and stable isotope labeled DPHM ([2H10]DPHM) via the umbilical vein (test route) and abdominal inferior vena cava (control route) in fetal lambs by either bolus injection (N = 6) or 90-min infusion (N = 5), and in nonpregnant sheep (N = 5), by simultaneous and separate bolus injections of the two forms of the drug via the mesenteric vein (test route) and abdominal inferior vena cava (control route). With the bolus injection protocol, hepatic presystemic elimination was estimated by the ratio of the area under the arterial drug concentration vs. time curve for the test and control routes of drug administration, whereas with the fetal infusion study it was calculated as the difference in fetal DPHM clearance values for the test and control routes of administration. To test for isotope effects in the disposition of [2H10]DPHM in the ewe or fetus, both DPHM and [2H10]DPHM were simultaneously injected via the inferior vena cava; however, no isotope effects were noted. In the ewe, there was extensive (93.2 +/- 1.4%) presystemic elimination of DPHM (F 0.068 +/- 0.014). However, in the fetus, this did not occur with bolus drug injection (F = 1.10 +/- 0.07), nor were there differences in the fetal DPHM clearance values estimated from the tarsal and umbilical venous infusions. During the latter experiments, DPHM levels were higher in the femoral artery than in carotid artery, with both umbilical venous and inferior vena caval drug infusion, and this was opposite of the differences in the concentrations of glucose, lactate, and oxygen between these two vessels. Thus, there is extensive hepatic presystemic elimination of DPHM in adult sheep, but no evidence for this phenomenon in the fetus. Furthermore, the preferential distribution of umbilical venous blood to the upper body of the fetal lamb does not result in higher drug levels in ascending aortic blood.


The global impact of noncommunicable diseases: estimates and projections.
With the aging of populations in developing countries there is both a demographic and an epidemiological transition which affects the impact of chronic degenerative diseases on the health status of the populations. Demographic transition takes place in countries where there are effective programmes of disease control which allow for survival during the early years of childhood and adolescence. This results in an increase in life expectancy which places larger proportions of the population in the age range (60 years and older) in which chronic degenerative diseases become the major determinants of health status. Epidemiological transition in diseases may also be brought about by shifts in social and economic patterns which favour detrimental changes in risk factors for the chronic degenerative diseases. Such changes may include health-related behaviour which augments dietary consumption of fats and alcohol, increases obesity, increases smoking and decreases physical activity. Such changes in risk-factor levels increase the prevalence of chronic degenerative diseases which manifest themselves at later ages, and for which early preventive actions could be cost-effective. In order to illustrate the impact of both demographic and risk-factor effects, analyses are made of the impact of increases in life expectancy on cause-specific mortality in both developing and developed countries. It is shown that there is great similarity in the effect of major noncommunicable diseases on the life expectancy of adults in both developed and developing countries. The major differences are seen to be in the proportions of deaths expected from such diseases as cancer, diabetes, heart disease, stroke and cirrhosis; but not in the distribution of age at death which is the better measure of disease impact. Demographic analyses, computing indirect estimates of mortality, also demonstrate that there are currently more chronic disease deaths in developing than developed countries and that as expectation of life increases in developing countries the global chronic disease burden will be greatly concentrated in the developing countries. Analyses of risk-factor reduction by feasible intervention strategies, e.g. smoking cessation campaigns, treatment of high blood pressure, using relationships between risk factors and diseases established in longitudinal studies carried out in developed countries, point out that the effect of risk-factor control in long-living populations can be hidden by the dependency of risk factors and various related causes of death, e.g. smoking has an impact on lung cancer, ischaemic heart disease and emphysema, but at different ages.(ABSTRACT TRUNCATED AT 400 WORDS)


Imatinib mesylate inhibits antigen-specific memory CD8 T cell responses in vivo.
Imatinib mesylate (IM) is effective at inducing complete cytogenetic remission in patients with chronic myelogenous leukemia. Because its influence on CD8 T cell responsiveness in vivo is unknown, we investigated the effects of IM by analyzing the response of OT-1 CD8 T cells to Listeria monocytogenes (LM) that express the cognate epitope OVA(257-264) (LM-OVA). In vitro, IM had no effect on Ag-specific expansion, cell division, cell cycle progression, or IFN-gamma expression in naive or memory OT-1 T cells. However, IM induced apoptosis of naive and memory OT-1 T cells at doses of >5 microM. At 15 microM IM, OT-1 T cells did not survive in in vitro cultures. The primary response of OT-1 T cells in vivo to LM-OVA infection was unaltered. In contrast, continuous IM treatment resulted in a diminished memory OT-1 response. The expression of IL-7Ralpha, a receptor required for memory cell survival, was lower (on OT-1 cells) in animals receiving IM. These results indicate that IM treatment affects the ability of the CD8 memory pool to respond to Ag and has the potential to increase susceptibility to infection.


Highly virulent Leptospira borgpetersenii strain characterized in the hamster model.
Abstract. A recent study by our group reported the isolation and partial serological and molecular characterization of four Leptospira borgpetersenii serogroup Ballum strains. Here, we reproduced experimental leptospirosis in golden Syrian hamsters (Mesocricetus auratus) and carried out standardization of lethal dose 50% (LD50) of one of these strains (4E). Clinical disease features and histopathologic analyses of tissue lesions were also observed. As results, strain 4E induced lethality in the hamster model with inocula lower than 10 leptospires, and histopathological examination of animals showed typical lesions found in severe leptospirosis. Gross pathological findings were peculiar; animals that died early had more chance of presenting severe jaundice and less chance of presenting pulmonary hemorrhages (P < 0.01). L. borgpetersenii serogroup Ballum has had a considerable growth in human leptospirosis cases in recent years. This strain has now been thoroughly characterized and can be used in more studies, especially evaluations of vaccine candidates.


'Over-servicing 'underservicing' and 'abandonment': What is the law?
The Ethical Rules and Policy Document of the Health Professions Council of South Africa (HPCSA) do not define 'over-servicing', 'underservicing' and 'abandonment. The HPCSA Guidelines on Over-servicing, Perverse Incentives and Related Matters define 'over-service' only. The converse of this definition can be used to define 'underservicing'. The courts do not refer to these concepts, but apply general rules regarding professional negligence and malpractice based on what a reasonably competent doctor in the same position would do. In deciding the standard to be adopted, the courts may consult the ethical rules of the medical profession, but are not bound to take them into account.


Design and evaluation of the LOPES exoskeleton robot for interactive gait rehabilitation.
This paper introduces a newly developed gait rehabilitation device. The device, called LOPES, combines a freely translatable and 2-D-actuated pelvis segment with a leg exoskeleton containing three actuated rotational joints: two at the hip and one at the knee. The joints are impedance controlled to allow bidirectional mechanical interaction between the robot and the training subject. Evaluation measurements show that the device allows both a "patient-in-charge" and "robot-in-charge" mode, in which the robot is controlled either to follow or to guide a patient, respectively. Electromyography (EMG) measurements (one subject) on eight important leg muscles, show that free walking in the device strongly resembles free treadmill walking; an indication that the device can offer task-specific gait training. The possibilities and limitations to using the device as gait measurement tool are also shown at the moment position measurements are not accurate enough for inverse-dynamical gait analysis.


An automatic method for robust and fast cell detection in bright field images from high-throughput microscopy.
In recent years, high-throughput microscopy has emerged as a powerful tool to analyze cellular dynamics in an unprecedentedly high resolved manner. The amount of data that is generated, for example in long-term time-lapse microscopy experiments, requires automated methods for processing and analysis. Available software frameworks are well suited for high-throughput processing of fluorescence images, but they often do not perform well on bright field image data that varies considerably between laboratories, setups, and even single experiments. In this contribution, we present a fully automated image processing pipeline that is able to robustly segment and analyze cells with ellipsoid morphology from bright field microscopy in a high-throughput, yet time efficient manner. The pipeline comprises two steps: (i) Image acquisition is adjusted to obtain optimal bright field image quality for automatic processing. (ii) A concatenation of fast performing image processing algorithms robustly identifies single cells in each image. We applied the method to a time-lapse movie consisting of ∼315,000 images of differentiating hematopoietic stem cells over 6 days. We evaluated the accuracy of our method by comparing the number of identified cells with manual counts. Our method is able to segment images with varying cell density and different cell types without parameter adjustment and clearly outperforms a standard approach. By computing population doubling times, we were able to identify three growth phases in the stem cell population throughout the whole movie, and validated our result with cell cycle times from single cell tracking. Our method allows fully automated processing and analysis of high-throughput bright field microscopy data. The robustness of cell detection and fast computation time will support the analysis of high-content screening experiments, on-line analysis of time-lapse experiments as well as development of methods to automatically track single-cell genealogies.


Toxicity and intracellular accumulation of bile acids in sandwich-cultured rat hepatocytes: role of glycine conjugates.
Excessive intrahepatic accumulation of bile acids (BAs) is a key mechanism underlying cholestasis. The aim of this study was to quantitatively explore the relationship between cytotoxicity of BAs and their intracellular accumulation in sandwich-cultured rat hepatocytes (SCRH). Following exposure of SCRH (on day-1 after seeding) to various BAs for 24h, glycine-conjugated BAs were most potent in exerting toxicity. Moreover, unconjugated BAs showed significantly higher toxicity in day-1 compared to day-3 SCRH. When day-1/-3 SCRH were exposed (0.5-4h) to 5-100μM (C)DCA, intracellular levels of unconjugated (C)DCA were similar, while intracellular levels of glycine conjugates were up to 4-fold lower in day-3 compared to day-1 SCRH. Sinusoidal efflux was by far the predominant efflux pathway of conjugated BAs both in day-1 and day-3 SCRH, while canalicular BA efflux showed substantial interbatch variability. After 4h exposure to (C)DCA, intracellular glycine conjugate levels were at least 10-fold higher than taurine conjugate levels. Taken together, reduced BA conjugate formation in day-3 SCRH results in lower intracellular glycine conjugate concentrations, explaining decreased toxicity of (C)DCA in day-3 versus day-1 SCRH. Our data provide for the first time a direct link between BA toxicity and glycine conjugate exposure in SCRH.


Genetic effect of the Aegilops caudata plasmon on the manifestation of the Ae. cylindrica genome.
In the course of reconstructing Aegilops caudata from its own genome (CC) and its plasmon, which had passed half a century in common wheat (genome AABBDD), we produced alloplasmic Ae. cylindrica (genome CCDD) with the plasmon of Ae. caudata. This line, designated (caudata)-CCDD, was found to express male sterility in its second substitution backcross generation (SB2) of (caudata)-AABBCCDD pollinated three times with the Ae. cylindrica pollen. We repeatedly backcrossed these SB2 plants with the Ae. cylindrica pollen until the SB5 generation, and SB5F2 progeny were produced by self-pollination of the SB5 plants. Thirteen morphological and physiological characters, including pollen and seed fertilities, of the (caudata)-CCDD SB5F2 were compared with those of the euplasmic Ae. cylindrica. The results indicated that the male sterility expressed by (caudata)-CCDD was due to genetic incompatibility between the Ae. cylindrica genome and Ae. caudata plasmon that did not affect any other characters of Ae. cylindrica. Also, we report that the genome integrity functions in keeping the univalent transmission rate high.


25-hydroxyvitamin D status does not affect intraoperative parathyroid hormone dynamics in patients with primary hyperparathyroidism.
Deficiency of 25-hydroxyvitamin D (25OHD) is a stimulus for the secretion of parathyroid hormone (PTH). During surgery for primary hyperparathyroidism, 25OHD deficiency may artificially elevate PTH, decreasing the sensitivity of intraoperative PTH (ioPTH) measurements. Of 351 patients with known 25OHD status who underwent curative surgical treatment of primary hyperparathyroidism, 198 (56%) patients were 25OHD deficient (<25 ng/mL). A curative decrease in ioPTH was defined as a greater than 50% PTH decrease 5, 10, or 15 min after resection. There was no statistical difference between 25OHD deficient and sufficient patients in PTH, phosphorous, and alkaline phosphatase preoperatively. There was a positive correlation between PTH and calcium, alkaline phosphatase, and gland weight, while there was an inverse correlation between preoperative PTH and 25OHD. The average ioPTH decrease was not significantly different after 5, 10, or 15 min, and 25OHD status did not affect when ioPTH indicated surgical cure. Lower 25OHD levels are correlated with higher PTH and alkaline phosphatase levels in patients with primary hyperparathyroidism. 25OHD status did not affect the average percent ioPTH decrease or the rate of cure. 25OHD deficiency does not affect ioPTH dynamics.


The long road to semantic interoperability in support of public health: experiences from two states.
Proliferation of health information technologies creates opportunities to improve clinical and public health, including high quality, safer care and lower costs. To maximize such potential benefits, health information technologies must readily and reliably exchange information with other systems. However, evidence from public health surveillance programs in two states suggests that operational clinical information systems often fail to use available standards, a barrier to semantic interoperability. Furthermore, analysis of existing policies incentivizing semantic interoperability suggests they have limited impact and are fragmented. In this essay, we discuss three approaches for increasing semantic interoperability to support national goals for using health information technologies. A clear, comprehensive strategy requiring collaborative efforts by clinical and public health stakeholders is suggested as a guide for the long road towards better population health data and outcomes.


Diverse antiapoptotic signaling pathways activated by vasoactive intestinal polypeptide, epidermal growth factor, and phosphatidylinositol 3-kinase in prostate cancer cells converge on BAD.
It has been demonstrated that vasoactive intestinal polypeptide, epidermal growth factor, and chronic activation of phosphatidylinositol 3-kinase can protect prostate cancer cells from apoptosis; however, the signaling pathways that they use and molecules that they target are unknown. We report that vasoactive intestinal polypeptide, epidermal growth factor, and phosphatidylinositol 3-kinase activate independent signaling pathways that phosphorylate the proapoptotic protein BAD. Vasoactive intestinal polypeptide operated via protein kinase A, epidermal growth factor required Ras activity, and effects of phosphatidylinositol 3-kinase were predominantly mediated by Akt. BAD phosphorylation was critical for the antiapoptotic effects of each signaling pathway. None of these survival signals was able to rescue cells that express BAD with mutations in phosphorylation sites, whereas knockdown of BAD expression with small hairpin RNA rendered cells insensitive to apoptosis. Taken together, these results identify BAD as a convergence point of several antiapoptotic signaling pathways in prostate cells.


Incomplete transient ischemia: a non-destructive evaluation of in vivo cerebral metabolism and hemodynamics in rat brain.
Differential near infrared spectrophotometry was used to monitor sequential in vivo alterations in cerebral hemoglobin saturation, blood volume, and cytochrome c oxidase reduction/oxidation responses during and after a period of incomplete transient ischemia (acute, reversible common carotid artery occlusion). In this study the rat brain was monitored non-invasively in the intact skull by transillumination. The data show that an increase in cerebral deoxygenation of hemoglobin, which occurs simultaneously with a decrease in blood volume subsequent to carotid ligation, acts as a compensatory mechanism to assist in maintaining aerobic energy metabolism. The observations also demonstrate that in this species the effects of bilateral carotid occlusion on the cerebrovascular parameters are not necessarily irrevocable. The intramitochondrial metabolic alterations, as evaluated by cytochrome c oxidase redox transitions, are reversible as long as the systemic arterial blood pressure does not fall below a value of approximately 40 mm Hg. These data suggest that possibility of being able to use a critical reduction level of cytochrome c oxidase as an early indication of ischemia-induced cerebral metabolic dysfunction prior to major changes in high energy stores.


Seasonal Dynamics of Phlebotomine Sand Fly Species Proven Vectors of Mediterranean Leishmaniasis Caused by Leishmania infantum.
The recent geographical expansion of phlebotomine vectors of Leishmania infantum in the Mediterranean subregion has been attributed to ongoing climate changes. At these latitudes, the activity of sand flies is typically seasonal; because seasonal phenomena are also sensitive to general variations in climate, current phenological data sets can provide a baseline for continuing investigations on sand fly population dynamics that may impact on future scenarios of leishmaniasis transmission. With this aim, in 2011-2013 a consortium of partners from eight Mediterranean countries carried out entomological investigations in sites where L. infantum transmission was recently reported. A common protocol for sand fly collection included monthly captures by CDC light traps, complemented by sticky traps in most of the sites. Collections were replicated for more than one season in order to reduce the effects of local weather events. In each site, the trapping effort was left unchanged throughout the survey to legitimate inter-seasonal comparisons. Data from 99,000 collected specimens were analyzed, resulting in the description of seasonal dynamics of 56,000 sand flies belonging to L. infantum vector species throughout a wide geographical area, namely P. perniciosus (Portugal, Spain and Italy), P. ariasi (France), P. neglectus (Greece), P. tobbi (Cyprus and Turkey), P. balcanicus and P. kandelakii (Georgia). Time of sand fly appearance/disappearance in collections differed between sites, and seasonal densities showed variations in each site. Significant correlations were found between latitude/mean annual temperature of sites and i) the first month of sand fly appearance, that ranged from early April to the first half of June; ii) the type of density trend, varying from a single peak in July/August to multiple peaks increasing in magnitude from May through September. A 3-modal trend, recorded for P. tobbi in Cyprus, represents a novel finding for a L. infantum vector. Adults ended the activity starting from mid September through November, without significant correlation with latitude/mean annual temperature of sites. The period of potential exposure to L.infantum in the Mediterranean subregion, as inferred by adult densities calculated from 3 years, 37 sites and 6 competent vector species, was associated to a regular bell-shaped density curve having a wide peak center encompassing the July-September period, and falling between early May to late October for more than 99% of values. Apparently no risk for leishmaniasis transmission took place from December through March in the years considered. We found a common pattern of nocturnal females activity, whose density peaked between 11 pm and 2 am. Despite annual variations, multiple collections performed over consecutive years provided homogeneous patterns of the potential behavior of leishmaniasis vectors in selected sites, which we propose may represent sentinel areas for future monitoring. In the investigated years, higher potential risk for L. infantum transmission in the Mediterranean was identified in the June-October period (97% relative vector density), however such risk was not equally distributed throughout the region, since density waves of adults occurred earlier and were more frequent in southern territories.


The use, appropriateness and outcomes of outpatient parenteral antimicrobial therapy.
Outpatient parenteral antimicrobial therapy (OPAT) is increasingly used to treat children at home, but studies in children are scarce. We aimed to describe the use, appropriateness and outcomes of OPAT in children. This was a 12-month prospective observational study. The hospital-in-the-home programme of The Royal Children's Hospital Melbourne. All patients receiving OPAT. Data were collected including demographics, diagnosis, type of venous access and antibiotic choice. Length of stay, adverse events, readmission rate and appropriateness of antibiotic use. 228 patients received OPAT in 251 episodes. The median age was 7.4 years (range 1 week to 21 years), with 22 patients (10%) under 1 year. The most frequent diagnoses were exacerbation of cystic fibrosis (17%), urinary tract infection (12%) and cellulitis (9%). Most patients were transferred from the ward, but 18% were transferred directly from the emergency department, the majority with skin and soft-tissue infection (66%). Venous access was most commonly peripherally inserted central catheter (29%) and peripheral cannula (29%). 309 parenteral antibiotics were prescribed, most frequently ceftriaxone (28%) and gentamicin (19%). The majority of antibiotics (72%) were prescribed appropriately. However, 6% were deemed an inappropriate choice for the indication and 26% had inappropriate dose or duration. The incidence of central line-associated bloodstream infections was 0.9%. The unplanned readmission rate was 4%, with low rates of OPAT-related adverse events. Three children (1%) had an inadequate clinical response. OPAT is a safe and effective way of providing antibiotics to children. Despite high rates of appropriate antibiotic use, improvements can still be made.


Stress and the transformation of lymphocytes by Epstein-Barr virus.
Although various stressors appear to influence herpesvirus infections, the underlying mechanisms have not been studied. A prospective design was used to examine the effects of examination stress and loneliness on the transformation of B lymphocytes in mixed cultures of T and B lymphocytes by Epstein-Barr virus (EBV). Three blood samples were drawn from 42 EBV-seropositive medical students, with the baseline sample taken 1 month before their final examinations, the stress sample drawn on the first day of final examinations, and the third sample taken the first week after their return from summer vacation. A median split on the UCLA Loneliness Scale divided the subjects into high- and low-scoring loneliness groups. There were significant effects for change over trials, with the lowest transformation levels (i.e., more virus required to transform cells) found in the stress sample. There was also a significant main effect for loneliness, in which high loneliness was associated with lower transformation levels. Possible immunological pathways for the observed changes are discussed.


Effectiveness of interventions designed to reduce the use of imaging for low-back pain: a systematic review.
Rates of imaging for low-back pain are high and are associated with increased health care costs and radiation exposure as well as potentially poorer patient outcomes. We conducted a systematic review to investigate the effectiveness of interventions aimed at reducing the use of imaging for low-back pain. We searched MEDLINE, Embase, CINAHL and the Cochrane Central Register of Controlled Trials from the earliest records to June 23, 2014. We included randomized controlled trials, controlled clinical trials and interrupted time series studies that assessed interventions designed to reduce the use of imaging in any clinical setting, including primary, emergency and specialist care. Two independent reviewers extracted data and assessed risk of bias. We used raw data on imaging rates to calculate summary statistics. Study heterogeneity prevented meta-analysis. A total of 8500 records were identified through the literature search. Of the 54 potentially eligible studies reviewed in full, 7 were included in our review. Clinical decision support involving a modified referral form in a hospital setting reduced imaging by 36.8% (95% confidence interval [CI] 33.2% to 40.5%). Targeted reminders to primary care physicians of appropriate indications for imaging reduced referrals for imaging by 22.5% (95% CI 8.4% to 36.8%). Interventions that used practitioner audits and feedback, practitioner education or guideline dissemination did not significantly reduce imaging rates. Lack of power within some of the included studies resulted in lack of statistical significance despite potentially clinically important effects. Clinical decision support in a hospital setting and targeted reminders to primary care doctors were effective interventions in reducing the use of imaging for low-back pain. These are potentially low-cost interventions that would substantially decrease medical expenditures associated with the management of low-back pain.


Evaluating farmers' adaptation strategies to climate change: A case study of Kaou local government area, Tahoua State, Niger Republic.
The study discusses local farmers' strategies of coping with and building resilience against the negative impact of climate change. Information for the discussion was from data collected using a set of structured questionnaires from interviews scheduled with 128 farmers. The questionnaire was based on previous literature and direct reconnaissance interview with farmers, which culminated in 13 strategies used for the study being reported. For each question, respondents were asked to choose their best and worst strategies. Thus, the difference between the best and worst strategies consistent with random utility theory has been used for the modelling. Results show that semi-transhumance, various handicrafts making, rural migration, small-scale vegetable production and small-scale river exploitation were the most important strategies identified, whilst water transport and vending, shifting cultivation, gypsum mining, gathering and trading of wild fruits and edible plants as well as cattle and sheep fattening were the least appreciated strategies identified amongst the farmers facing climate change. These findings are therefore imperative for planning farmers' capacity-building and resilience against climate change projects to ensure sustainability in the study area.


NMR Study of Ion Dynamics and Charge Storage in Ionic Liquid Supercapacitors.
Ionic liquids are emerging as promising new electrolytes for supercapacitors. While their higher operating voltages allow the storage of more energy than organic electrolytes, they cannot currently compete in terms of power performance. More fundamental studies of the mechanism and dynamics of charge storage are required to facilitate the development and application of these materials. Here we demonstrate the application of nuclear magnetic resonance spectroscopy to study the structure and dynamics of ionic liquids confined in porous carbon electrodes. The measurements reveal that ionic liquids spontaneously wet the carbon micropores in the absence of any applied potential and that on application of a potential supercapacitor charging takes place by adsorption of counterions and desorption of co-ions from the pores. We find that adsorption and desorption of anions surprisingly plays a more dominant role than that of the cations. Having elucidated the charging mechanism, we go on to study the factors that affect the rate of ionic diffusion in the carbon micropores in an effort to understand supercapacitor charging dynamics. We show that the line shape of the resonance arising from adsorbed ions is a sensitive probe of their effective diffusion rate, which is found to depend on the ionic liquid studied, as well as the presence of any solvent additives. Taken as whole, our NMR measurements allow us to rationalize the power performances of different electrolytes in supercapacitors.


DNA looping and unlooping by AraC protein.
Expression of the L-arabinose BAD operon in Escherichia coli is regulated by AraC protein which acts both positively in the presence of arabinose to induce transcription and negatively in the absence of arabinose to repress transcription. The repression of the araBAD promoter is mediated by DNA looping between AraC protein bound at two sites near the promoter separated by 210 base pairs, araI and araO2. In vivo and in vitro experiments presented here show that an AraC dimer, with binding to half of araI and to araO2, maintains the repressed state of the operon. The addition of arabinose, which induces the operon, breaks the loop, and shifts the interactions from the distal araO2 site to the previously unoccupied half of the araI site. The conversion between the two states does not require additional binding of AraC protein and appears to be driven largely by properties of the protein rather than being specified by the slightly different DNA sequences of the binding sites. Slight reorientation of the subunits of AraC could specify looping or unlooping by the protein. Such a mechanism could account for regulation of DNA looping in other systems.


Diagnosis and treatment of vitamin D deficiency.
The recent discovery--in a randomised, controlled trial--that daily ingestion of 1100 IU of colecalciferol (vitamin D) over a 4-year period dramatically reduced the incidence of non-skin cancers makes it difficult to overstate the potential medical, social and economic implications of treating vitamin D deficiency. Not only are such deficiencies common, probably the rule, vitamin D deficiency stands implicated in a host of diseases other than cancer. The metabolic product of vitamin D is a potent, pleiotropic, repair and maintenance, secosteroid hormone that targets > 200 human genes in a wide variety of tissues, meaning it has as many mechanisms of action as genes it targets. A common misconception is that government agencies designed present intake recommendations to prevent or treat vitamin D deficiency. They did not. Instead, they are guidelines to prevent particular metabolic bone diseases. Official recommendations were never designed and are not effective in preventing or treating vitamin D deficiency and in no way limit the freedom of the physician--or responsibility--to do so. At this time, assessing serum 25-hydroxy-vitamin D is the only way to make the diagnosis and to assure that treatment is adequate and safe. The authors believe that treatment should be sufficient to maintain levels found in humans living naturally in a sun-rich environment, that is, > 40 ng/ml, year around. Three treatment modalities exist: sunlight, artificial ultraviolet B radiation or supplementation. All treatment modalities have their potential risks and benefits. Benefits of all treatment modalities outweigh potential risks and greatly outweigh the risk of no treatment. As a prolonged 'vitamin D winter', centred on the winter solstice, occurs at many temperate latitudes, < or = 5000 IU (125 microg) of vitamin D/day may be required in obese, aged and/or dark-skinned patients to maintain adequate levels during the winter, a dose that makes many physicians uncomfortable.


Sensitivity analyses for effect modifiers not observed in the target population when generalizing treatment effects from a randomized controlled trial: Assumptions, models, effect scales, data scenarios, and implementation details.
Randomized controlled trials are often used to inform policy and practice for broad populations. The average treatment effect (ATE) for a target population, however, may be different from the ATE observed in a trial if there are effect modifiers whose distribution in the target population is different that from that in the trial. Methods exist to use trial data to estimate the target population ATE, provided the distributions of treatment effect modifiers are observed in both the trial and target population-an assumption that may not hold in practice. The proposed sensitivity analyses address the situation where a treatment effect modifier is observed in the trial but not the target population. These methods are based on an outcome model or the combination of such a model and weighting adjustment for observed differences between the trial sample and target population. They accommodate several types of outcome models: linear models (including single time outcome and pre- and post-treatment outcomes) for additive effects, and models with log or logit link for multiplicative effects. We clarify the methods' assumptions and provide detailed implementation instructions. We illustrate the methods using an example generalizing the effects of an HIV treatment regimen from a randomized trial to a relevant target population. These methods allow researchers and decision-makers to have more appropriate confidence when drawing conclusions about target population effects.


Trends of surgical career selection among medical students and graduates: a global perspective.
Declining trends in surgery have been well-researched in the North American region, but little is known about the international dynamics affecting this phenomenon. A comprehensive approach to this emerging issue is lacking. Our objective in this study is to explore global trends related to medical student interest in surgical careers. We will also identify factors influencing the choice of surgery as a career and outline practical interventions to nurture interest in these fields, as described in the literature. We present a focused review of worldwide trends of surgical career selection among medical students of geographically distant and diverse sociocultural backgrounds. A systematic literature search was performed using PubMed. The studies were conducted over the last fifteen years with the study populations scattered over 6 continents. North American studies have shown a declining trend for medical students to select surgery as a career, but a recent recovery has been observed. However, surgery is still the popular choice of career among Middle Eastern and Far-Eastern Asian men. Sociocultural variants and personal satisfaction are likely influential factors with financial rewards being the least significant. Surgery is also popular in Nigeria despite their increased awareness of risk of HIV/AIDS associated with surgery. A decline in surgery has been observed in Australia, New Zealand, and South Africa. Medical students and graduates of Greece, Switzerland, and the UK mostly prefer nonsurgical careers. Multiple extrinsic and intrinsic factors appear to play a role for such a phenomenon. Specific interventions are now being used to create a rebound effect for such a declining trend observed in most of the regions worldwide. Global trends show a variation of interest in surgery as a career option throughout the 6 continents. Possible factors affecting this phenomenon are similar among geographically distant and diverse student cultures.


Sirt1 activation protects the mouse renal medulla from oxidative injury.
Sirtuin 1 (Sirt1) is a NAD+-dependent deacetylase that exerts many of the pleiotropic effects of oxidative metabolism. Due to local hypoxia and hypertonicity, the renal medulla is subject to extreme oxidative stress. Here, we set out to investigate the role of Sirt1 in the kidney. Our initial analysis indicated that it was abundantly expressed in mouse renal medullary interstitial cells in vivo. Knocking down Sirt1 expression in primary mouse renal medullary interstitial cells substantially reduced cellular resistance to oxidative stress, while pharmacologic Sirt1 activation using either resveratrol or SRT2183 improved cell survival in response to oxidative stress. The unilateral ureteral obstruction (UUO) model of kidney injury induced markedly more renal apoptosis and fibrosis in Sirt1+/- mice than in wild-type controls, while pharmacologic Sirt1 activation substantially attenuated apoptosis and fibrosis in wild-type mice. Moreover, Sirt1 deficiency attenuated oxidative stress-induced COX2 expression in cultured mouse renal medullary interstitial cells, and Sirt1+/- mice displayed reduced UUO-induced COX2 expression in vivo. Conversely, Sirt1 activation increased renal medullary interstitial cell COX2 expression both in vitro and in vivo. Furthermore, exogenous PGE2 markedly reduced apoptosis in Sirt1-deficient renal medullary interstitial cells following oxidative stress. Taken together, these results identify Sirt1 as an important protective factor for mouse renal medullary interstitial cells following oxidative stress and suggest that the protective function of Sirt1 is partly attributable to its regulation of COX2 induction. We therefore suggest that Sirt1 provides a potential therapeutic target to minimize renal medullary cell damage following oxidative stress.


Secondhand smoke exposure among nonsmoking adults in two Nigerian cities.
Tobacco control policy can only succeed if the burdens of smoking are known. The objective of this study was to determine the prevalence and correlates of secondhand smoke (SHS) exposure among nonsmoking adults in two Nigerian cities. We carried out a cross-sectional study from October 2009 to April 2010 among adult population of two Nigerian cities: Enugu and Ilorin. A semi-structured questionnaire was administered by interviewers to obtain socio-demographic information; and information regarding pattern of SHS exposure, awareness of tobacco control policy and the harmful effects of SHS. SHS exposure was defined as regular exposure to tobacco smoke in the previous 30 days in a nonsmoking adult. Of the 585 nonsmoking adults that completed the study, 38.8% had regular exposure to SHS; mostly, in public places (24.4%). More men were exposed at public places when compared with women (27.0% vs. 19.5%). The strongest factor associated with exposure to SHS in women was having a smoking spouse [prevalence rate (PR) ratio-7.76; 95% confidence interval (CI), 3.08-9.42]; and in men, it was lack of home smoking restriction (PR ratio-6.35; 95% CI, 4.51-8.93). Among men, SHS exposure at any location was associated with lack of secondary school education, residing in slum apartment (house with many households), living with a smoking family member (non-spouse), lack of home smoking restriction, and alcohol intake. Among women, SHS exposure at any location was associated with having a smoking spouse, residing in slum apartment and lack of home smoking restriction. Seventy-two percent of respondents were aware of the harmful effects of SHS on their health. Lack of awareness of the harmful effects was significantly associated with increasing age (r = +0.45; P = <0.01), lack of secondary school education (r = -0.10; P = 0.04), residing in slum apartment (r = -0.12; P = 0.03) and being a widow/widower (r = +0.24; P < 0.01). Only 17.4% of the employees reported availability of outdoor smoking area at their workplaces. Our results show that prevalence of SHS exposure was the highest in public places. These findings underscore the need for enactment of comprehensive smoke-free legislation and implementation of educational strategies to reduce SHS exposure in homes.


Catalysis of strand exchange by the HSV-1 UL12 and ICP8 proteins: potent ICP8 recombinase activity is revealed upon resection of dsDNA substrate by nuclease.
The replication of herpes simplex virus type 1 (HSV-1) is associated with a high degree of homologous recombination, which is likely to be mediated, in part, by HSV-1-encoded proteins. We have previously shown that the HSV-1 encoded ICP8 protein and alkaline nuclease UL12 are capable of catalyzing an in vitro strand-exchange reaction. Here, we show, by electron microscopy, that the products of the strand exchange reaction between linear double-stranded DNA and circular single-stranded DNA consist of the expected joint molecule forms: sigma, alpha, and gapped circles. Other exonucleases, such as lambda Red alpha, which, like UL12, digests 5'-3', as well as Escherichia coli exonuclease III (ExoIII), which digests 3'-5', could substitute for UL12 in the strand exchange reaction by providing a resected DNA end. ICP8 generated the same intermediates and strand exchange products when the double-stranded DNA substrate was preresected by any of the nucleases. Using substrates with large regions of non-homology we found that pairing by ICP8 could be initiated from the middle of a DNA molecule and did not require a homologous end. In this reaction, the resection of a DNA end by the nuclease is required to reveal homologous sequences capable of being paired by ICP8. This study further illustrates the complexity of the multi-functional ICP8 protein.


Continuing drug risk behaviour: shared use of injecting paraphernalia among London heroin injectors.
The study investigates donor sharing and recipient sharing of different types of injecting equipment and the frequency of sharing activities with sexual partners, close friends and casual acquaintances. Structured interviews were conducted with 303 injectors recruited as part of a study of early and episodic opiate use. Subjects were contacted in non-clinical, community settings by a peer recruitment method (Privileged Access Interviews). Spoons and water containers were more frequently shared than needles and syringes. Significant differences were obtained between the frequency of sharing different types of injecting equipment. Many injectors had shared spoons or water containers, but had not shared needles or syringes. Frequency of sharing with others was also associated with intimacy of relationship to the subject. Almost two-thirds of the sample had shared some sort of injecting equipment during the previous year. The sharing of injecting paraphernalia presents a risk of infection with HIV hepatitis B and hepatitis C. Injectors and drug workers should be more clearly aware of the risks involved in such activities. Harm reduction advice given to drug injectors should target the sharing of injecting paraphernalia. Research into sharing should explicitly include questions about the use of different types of injecting equipment.


Detection of fusion transcripts generated by the inversion 16 chromosome in acute myelogenous leukemia.
Pericentric inversion of chromosome 16 [inv(16)(p13q22)] and the related t(16;16)(p13;q22) are seen in a subset of acute myelogenous leukemia (AML) phenotypically and prognostically differing from other cases. We have recently shown that inv(16) results in fusion of CBFB/PEBP2B, a gene encoded at 16q22 to MYH11, a smooth muscle myosin heavy chain gene encoded at 16p13. Chimeric transcripts consisting of upstream CBFB fused to downstream MYH11 coding sequences result from this fusion. In this study we have examined a series of 37 of these cases using reverse transcriptase-polymerase chain reaction (RT-PCR) to detect expression of a hybrid CBFB/MYH11 transcript. Chimeric cDNAs were detected in all but 1 of 37 leukemias with typical inv(16) or t(16;16). Such chimeric products were not seen in a case with inv(16)(p13q24) (ie, a variant q arm breakpoint) or any of 10 cases of AML without these chromosomal changes. Four different chimeric transcripts were found, representing differing fusion points within MYH11 spliced to position 495 of CBFB. Primer sets are described for efficient amplification of these different cDNA forms. Amplification of cDNA showed that all but 17 codons of the CBFB coding sequence are included in the abnormal transcripts. RT-PCR was shown to be highly sensitive and potentially useful for detection of leukemic cells during morphologic remission.


Structural and spectroscopic trends in a series of half-sandwich scorpionate complexes.
Fifteen half-sandwich scorpionate complexes [(L)M(NCMe)(3)](BF(4))(n) (L = tris(3,5-dimethylpyrazol-1-yl)methane, Tpm(Me,Me), n = 2, 1(M), M = Mn, Fe, Co, Ni; L = tris(3-phenylpyrazol-1-yl)methane, Tpm(Ph), n = 2, 2(M), M = Mn, Fe, Co, Ni; L = hydrotris(3,5-dimethylpyrazol-1-yl)borate, [Tp(Me,Me)](-), n = 1, 3(M), M = Fe, Co, Ni; L = hydrotris(3-phenyl-5-methylpyrazol-1-yl)borate, [Tp(Ph,Me)](-), n = 1, 4(M), M = Mn, Fe, Co, Ni) were prepared by addition of the tripodal ligands to solvated [M(NCMe)(x)](2+) (M = Mn, x = 4; M = Fe, Co, Ni, x = 6) precursor complexes. The product complexes were characterized by (1)H NMR (except M = Mn), UV-vis-NIR, and FTIR spectroscopy. The structures of 2(Mn), 2(Ni), 3(Fe), 3(Co), and 4(Fe) were determined by X-ray crystallography. The data were consistent with complexes of high-spin divalent metal ions in idealized piano-stool geometries in all cases. Consequent lability of the acetonitrile ligands will enable use of these complexes as synthetic precursors and as catalysts. Comparison to previously reported structures of 1(Fe), 1(Co), 2(Fe), and 2(Co), the triflate salt analogues of 4(Co) and 4(Ni), as well as related sandwich complexes (e.g., [(Tp(Me,Me))(2)M]) and solvated metal dications [M(NCMe)(6)](2+) reveals numerous trends in M-N bond lengths. Primary among these are the Irving-Williams series, with significant structural effects also arising from ligand charge and sterics. Systematic trends in spectroscopic data were also observed which further elucidate these issues.


Molecular cloning and transcriptional and expression analysis of engO, encoding a new noncellulosomal family 9 enzyme, from Clostridium cellulovorans.
Clostridium cellulovorans produces a major noncellulosomal family 9 endoglucanase EngO. A genomic DNA fragment (40 kb) containing engO and neighboring genes was cloned. The nucleotide sequence contained reading frames for endoglucanase EngO, a putative response regulator, and a putative sensor histidine kinase protein. The engO gene consists of 2,172 bp and encodes a protein of 724 amino acids with a molecular weight of 79,474. Northern hybridizations revealed that the engO gene is transcribed as a monocistronic 2.6-kb mRNA. 5' RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) PCR analysis indicated that the single transcriptional start site of engO was located 264 bp upstream from the first nucleotide of the translation initiation codon. Alignment of the engO promoter region provided evidence for highly conserved sequences that exhibited strong similarity to the sigma(A) consensus promoter sequences of gram-positive bacteria. EngO contains a typical N-terminal signal peptide of 28 amino acid residues, followed by a 149-amino-acid sequence which is homologous to the family 4-9 carbohydrate-binding domain. Downstream of this domain was an immunoglobulin-like domain of 89 amino acids. The C terminus contains a family 9 catalytic domain of glycosyl hydrolase. Mass spectrometry analysis of EngO was in agreement with that deduced from the nucleotide sequence. Expression of engO mRNA increased from early to middle exponential phase and decreased during the early stationary phase. EngO was highly active toward carboxymethyl cellulose but showed no activity towards xylan. It was optimally active at 40 to 50 degrees C and pH 5 to 6. The analysis of the products from the cellulose hydrolysis through thin-layer chromatography indicated its endoglucanase activity.


MicroRNAs in rheumatoid arthritis: potential role in diagnosis and therapy.
Rheumatoid arthritis (RA) is a systemic, inflammatory, autoimmune disorder with progressive articular damage that may result in lifelong disability. Although major strides in understanding the disease have been made, the pathogenesis of RA has not yet been fully elucidated. Early treatment can prevent severe disability and lead to remarkable patient benefits, although a lack of therapeutic efficiency in a considerable number of patients remains problematic. MicroRNAs (miRNAs) are small, non-coding RNAs that, depending upon base pairing to messenger RNA (mRNA), mediate mRNA cleavage, translational repression or mRNA destabilization. As fine tuning regulators of gene expression, miRNAs are involved in crucial cellular processes and their dysregulation has been described in many cell types in different diseases. In body fluids, miRNAs are present in microvesicles or incorporated into complexes with Argonaute 2 (Ago2) or high-density lipoproteins and show high stability. Therefore, they are of interest as potential biomarkers of disease in daily diagnostic applications. Targeting miRNAs by gain or loss of function approaches have brought therapeutic effects in various animal models. Over the past several years it has become clear that alterations exist in the expression of miRNAs in patients with RA. Increasing numbers of studies have shown that dysregulation of miRNAs in peripheral blood mononuclear cells or isolated T lymphocytes, in synovial tissue and synovial fibroblasts that are considered key effector cells in joint destruction, contributes to inflammation, degradation of extracellular matrix and invasive behaviour of resident cells. Thereby, miRNAs maintain the pathophysiological process typical of RA. The aim of the current review is to discuss the available evidence linking the expression of miRNAs to inflammatory and immune response in RA and their potential as biomarkers and the novel targets for treatment in patients with RA.


Development and validation of a nomogram to individually predict survival of young patients with nonmetastatic gastric cancer: A retrospective cohort study.
Evidence regarding gastric cancer (GC) patients <40 years old is limited. The aim of the study was to identify risk factors affecting overall survival (OS) of young patients with nonmetastatic GC and to establish a nomogram for prognostic prediction using data from the Surveillance, Epidemiology and End Results (SEER) database. Furthermore, this study sought to externally validate this nomogram in an independent patient cohort. In this retrospective cohort study, the records of patients aged <40 years with nonmetastatic GC (n = 559), from the SEER database, between 2006 and 2015, were examined. The nomogram was established based on the Cox proportional hazards regression model using the SEER dataset. Patients with nonmetastatic GC (n = 201) in our department between 2009 and 2015 were selected as an external validation set. Discrimination and calibration were performed in both cohorts. The multivariate Cox model identified race, tumor subsites, tumor size, depth of invasion, lymph node metastasis, number of examined lymph nodes, and surgery as independent covariates associated with OS. The nomogram exhibited superior discriminative power than the eighth tumor, node, metastasis (TNM) staging system in both the training set [Harrell's concordance index (C index): 0.762 vs. 0.635,P < 0.001] and validation set (C index: 0.805 vs. 0.712,P= 0.176). Calibration of the nomogram was good in both cohorts. We developed a nomogram predicting 3- and 5-year OS rates in young patients with nonmetastatic GC. Both the training set and validation set showed good discrimination and calibration, suggesting good clinical applicability.


Genetic investigation of chromosome 5q GABAA receptor subunit genes in schizophrenia.
We previously performed a genome-wide linkage scan in Portuguese schizophrenia families that identified a risk locus on chromosome 5q31-q35. This finding was supported by meta-analysis of 20 other schizophrenia genome-wide scans that identified 5q23.2-q34 as the second most compelling susceptibility locus in the genome. In the present report, we took a two-stage candidate gene association approach to investigate a group of gamma-aminobutyric acid (GABA) A receptor subunit genes (GABRA1, GABRA6, GABRB2, GABRG2, and GABRP) within our linkage peak. These genes are plausible candidates based on prior evidence for GABA system involvement in schizophrenia. In the first stage, associations were detected in a Portuguese patient sample with single nucleotide polymorphisms (SNPs) and haplotypes in GABRA1 (P=0.00062-0.048), GABRP (P=0.0024-0.042), and GABRA6 (P=0.0065-0.0088). The GABRA1 and GABRP findings were replicated in the second stage in an independent German family-based sample (P=0.0015-0.043). Supportive evidence for association was also obtained for a previously reported GABRB2 risk haplotype. Exploratory analyses of the effects of associated GABRA1 haplotypes on transcript levels found altered expression of GABRA6 and coexpressed genes of GABRA1 and GABRB2. Comparison of transcript levels in schizophrenia patients and unaffected siblings found lower patient expression of GABRA6 and coexpressed genes of GABRA1. Interestingly, the GABRA1 coexpressed genes include synaptic and vesicle-associated genes previously found altered in schizophrenia prefrontal cortex. Taken together, these results support the involvement of the chromosome 5q GABAA receptor gene cluster in schizophrenia, and suggest that schizophrenia-associated haplotypes may alter expression of GABA-related genes.