Analysis of discrete local variability and structural covariance in macromolecular assemblies using Cryo-EM and focused classification.
Single-particle electron cryo-microscopy and computational image classification can be used to analyze structural variability in macromolecules and their assemblies. In some cases, a particle may contain different regions that each display a range of distinct conformations. We have developed strategies, implemented within the Frealign and cisTEM image processing packages, to focus-classify on specific regions of a particle and detect potential covariance. The strategies are based on masking the region of interest using either a 2-D mask applied to reference projections and particle images, or a 3-D mask applied to the 3-D volume. We show that focused classification approaches can be used to study structural covariance, a concept that is likely to gain more importance as datasets grow in size, allowing the distinction of more structural states and smaller differences between states. Finally, we apply the approaches to an experimental dataset containing the HIV-1 Transactivation Response (TAR) element RNA fused into the large bacterial ribosomal subunit to deconvolve structural mobility within localized regions of interest, and to a dataset containing assembly intermediates of the large subunit to measure structural covariance.


Molecular dissection of Penelope transposable element regulatory machinery.
Penelope-like elements (PLEs) represent a new class of retroelements identified in more than 80 species belonging to at least 10 animal phyla. Penelope isolated from Drosophila virilis is the only known transpositionally active representative of this class. Although the size and structure of the Penelope major transcript has been previously described in both D. virilis and D. melanogaster transgenic strains, the architecture of the Penelope regulatory region remains unknown. In order to determine the localization of presumptive Penelope promoter and enhancer-like elements, segments of the putative Penelope regulatory region were linked to a CAT reporter gene and introduced into D. melanogaster by P-element-mediated transformation. The results obtained using ELISA to measure CAT expression levels and RNA studies, including RT-PCR, suggest that the active Penelope transposon contains an internal promoter similar to the TATA-less promoters of LINEs. The results also suggest that some of the Penelope regulatory sequences control the preferential expression in the ovaries of the adult flies by enhancing expression in the ovary and reducing expression in the carcass. The possible significance of the intron within Penelope for the function and evolution of PLEs, and the effect of Penelope insertions on adjacent genes, are discussed.


A dissociated glucocorticoid receptor modulator reduces airway hyperresponsiveness and inflammation in a mouse model of asthma.
The glucocorticoid receptor (GR) is a transcription factor able to support either target gene activation via direct binding to DNA or gene repression via interfering with the activity of various proinflammatory transcription factors. An improved therapeutic profile for combating chronic inflammatory diseases has been reported through selectively modulating the GR by only triggering its transrepression function. We have studied in this paper the activity of Compound A (CpdA), a dissociated GR modulator favoring GR monomer formation, in a predominantly Th2-driven asthma model. CpdA acted similarly to the glucocorticoid dexamethasone (DEX) in counteracting OVA-induced airway hyperresponsiveness, recruitment of eosinophils, dendritic cells, neutrophils, B and T cells, and macrophages in bronchoalveolar lavage fluid, lung Th2, Tc2, Th17, Tc17, and mast cell infiltration, collagen deposition, and goblet cell metaplasia. Both CpdA and DEX inhibited Th2 cytokine production in bronchoalveolar lavage as well as nuclear translocation of NF-κB and its subsequent recruitment onto the IκBα promoter in the lung. By contrast, DEX but not CpdA induces expression of the GR-dependent model gene MAPK phosphatase 1 in the lung, confirming the dissociative action of CpdA. Mechanistically, we demonstrate that CpdA inhibited IL-4-induced STAT6 translocation and that GR is essential for CpdA to mediate chemokine repression. In conclusion, we clearly show in this study the anti-inflammatory effect of CpdA in a Th2-driven asthma model in the absence of transactivation, suggesting a potential therapeutic benefit of this strategy.


Arthroscopic treatment of labral tears in femoroacetabular impingement: a comparative study of refixation and resection with a minimum two-year follow-up.
Labral tears are commonly associated with femoroacetabular impingement. We reviewed 151 patients (156 hips) with femoroacetabular impingement and labral tears who had been treated arthroscopically. These were subdivided into those who had undergone a labral repair (group 1) and those who had undergone resection of the labrum (group 2). In order to ensure the groups were suitably matched for comparison of treatment effects, patients with advanced degenerative changes (Tönnis grade > 2, lateral sourcil height < 2 mm and Outerbridge grade 4 changes in the weight-bearing area of the femoral head) were excluded, leaving 96 patients (101 hips) in the study. At a mean follow-up of 2.44 years (2 to 4), the mean modified Harris hip score in the labral repair group (group 1, 69 hips) improved from 60.2 (24 to 85) pre-operatively to 93.6 (55 to 100), and in the labral resection group (group 2, 32 hips) from 62.8 (29 to 96) pre-operatively to 88.8 (35 to 100). The mean modified Harris hip score in the labral repair group was 7.3 points greater than in the resection group (p = 0.036, 95% confidence interval 0.51 to 14.09). Labral detachments were found more frequently in the labral repair group and labral flap tears in the resection group. No patient in our study group required a subsequent hip replacement during the period of follow-up. This study shows that patients without advanced degenerative changes in the hip can achieve significant improvement in their symptoms after arthroscopic treatment of femoroacetabular impingement. Where appropriate, labral repair provides a superior result to labral resection.


Mutational analysis of the membrane-proximal cleavage site of L-selectin: relaxed sequence specificity surrounding the cleavage site.
L-selectin expression is regulated in part by membrane-proximal cleavage from the cell surface of leukocytes and L-selectin-transfected cells. The downregulation of L-selectin from the surface of neutrophils is speculated to be a process involved in the adhesion cascade leading to neutrophil recruitment to sites of inflammation. We previously reported that L-selectin is cleaved between Lys321 and Ser322 in a region that links the second short consensus repeat (SCR) and the transmembrane domain. We demonstrate that replacing this cleavage domain of L-selectin with the corresponding region of E-selectin prevents L-selectin shedding, as judged by inhibiting the generation of the 68-kD soluble and 6-kD transmembrane cleavage products of L-selectin. Unexpectedly, we found that point mutations of the cleavage site, as well as mutations of multiple conserved amino acids within the cleavage domain, do not significantly affect L-selectin shedding. However, short deletions of four or five amino acids in the L-selectin cleavage domain inhibit L-selectin downregulation. Mutations that appeared to inhibit L-selectin shedding resulted in higher levels of cell surface expression, consistent with a lack of apparent proteolysis from the cell membrane. One deletion mutant, I327 delta N332, retains the native cleavage site yet inhibits L-selectin proteolysis as well. Restoring the amino acids deleted between I327 and N332 with five alanine residues restores L-selectin proteolysis. Thus, the proteolytic processing of L-selectin appears to have a relaxed sequence specificity at the cleavage site, and it may depend on the physical length or other secondary structural characteristics of the cleavage domain.


Five-year clinical results of immediately loaded dental implants using mandibular overdentures.
The aim of this report is to present the clinical and patient-based outcomes of an immediate-loading protocol of TiUnite implants with mandibular overdentures in edentulous patients 5 years following initial placement. The study comprised two groups of edentulous patients. In the experimental group, 35 consecutively treated patients received 70 TiUnite implants that were loaded immediately, as well as 69 Branemark machined implants as a backup treatment. One patient received one Branemark implant. The control group comprised patients who were treated previously with conventional two-stage implant procedures, but were all case matched to the intervention group and served as a historical cohort. This group included 42 patients who received 111 Branemark implants. Both groups of patients were treated with overdentures that were supported with a standardized resilient bar mechanism. Clinical and patient-based outcomes in the immediate group were recorded for the first 5 years following the initial placement of implants and were measured at various stages of treatment using two questionnaires: the Denture Satisfaction Scale and the Oral Health Impact Profile (OHIP-20). Just over 98% of implants were found to be successful in both groups (Fisher exact test: P = 1.000). A statistically significant improvement in patients' total, mandibular, and functional satisfaction scores was found when comparing baseline data to the data obtained 5 years following loading in the experimental group (P < .001). There were no significant differences between the 1- and 5-year total, mandibular, and functional satisfaction scores, or between baseline and 5-year maxillary denture satisfaction scores. A statistically significant and positive correlation was found between baseline and 1-year maxillary satisfaction scores (P = .002). Any improvement in the patients' quality of life (QoL) was maintained during the first 5 years of loading. The results of this longitudinal study suggest that immediate loading of two dental implants by means of bar-retained mandibular overdentures is a predictable treatment option and leads to substantial improvement in patients' satisfaction and QoL. Importantly, this mirrors the outcomes found for patients subjected to the more commonly accepted two-stage implant procedure.


Day 5 inhibin B levels in a treatment cycle are predictive of IVF outcome.
Day 5 serum inhibin B during IVF treatment has been investigated as a predictor of outcome. A total of 54 women (< or = 39 years, normal menses and endocrine profiles) were treated with urinary gonadotrophins or recombinant FSH following pituitary down-regulation. Serum day 3 FSH in a preceding cycle was <8.5 IU/l. Plasma inhibin B, inhibin A and estradiol were determined after 4 days of gonadotrophin administration (day 5). Day 5 inhibin B was the most highly correlated with the number of mature follicles (>14 mm), oocytes retrieved and fertilized. Receiver operating characteristic analysis gave high accuracy for day 5 inhibin B in predicting ovarian response and indicated that a threshold of 400 pg/ml may be helpful in the decision as to whether to continue treatment. Women with <400 pg/ml (n = 16) had lower numbers of follicles, mature follicles, oocytes retrieved, fertilized and cleaved compared with those >400 pg/ml (n = 36) and this threshold gave a positive likelihood ratio of 30, 92.9% sensitivity, 95.0% specificity and 86.7% positive predictive value to detect poor ovarian response. Day 5 inhibin B was the best predictor of pregnancy (no live births and four cycles cancelled, low inhibin group; nine live births and no cancelled cycles, high inhibin group). Normogonadotrophic, normogonadal women with day 5 inhibin B <400 pg/ml in down-regulated cycles have a poor response to ovarian stimulation and are less likely to conceive compared with women with higher day 5 inhibin B.


Reporting Physical Activity: Perceptions and Practices of Australian Media Professionals.
Advocacy informed by scientific evidence is necessary to influence policy and planning to address physical inactivity. The mass media is a key arena for this advocacy. This study investigated the perceptions and practices of news media professionals reporting physical activity and sedentariness to inform strategic communication about these issues. We interviewed media professionals working for major television, radio, newspaper and online news outlets in Australia. The interviews explored understandings of physical activity and sedentariness, attributions of causality, assignment of responsibility, and factors affecting news reporting on these topics. Data were thematically analyzed using NVivo. Physical inactivity was recognized as pervasive and important, but tended to be seen as mundane and not newsworthy. Sedentariness was regarded as more novel than physical activity, and more likely to require organizational and environment action. Respondents identified that presenting these issues in visual and engaging ways was an ongoing challenge. Physical activity researchers and advocates need to take account of prevailing news values and media practices to improve engagement with the news media. These include understanding the importance of novelty, narratives, imagery, and practical messages, and how to use these to build support for environmental and policy action.


Positive life orientation--an inner health resource among older people.
As pertains to life orientation, the study's conceptual framework demonstrates the importance of close relationships, meaning in life, and the strength of individuals to look forward and make plans for future. The aim of this study is to discover, in relation to background factors such as language, living situation, and culture, how people aged 65 and 75 experience life orientation. The study is a population-based cross-sectional survey with its material taken from 3,370 questionnaires which included the six-question Life Orientation scale. The SPSS static program, version 14.0, was used for all calcuations and the Chi-squared test was applied to determine if any significant differences between the groups existed. As seen in this study, most respondents aged 65 (85%) and 75 (73.2%) experienced a high degree of positive life orientation, clearly expressing a zest for life (93.5%), overall satisfaction with their lives (88.4%), and feelings of being needed (83.7%). Lower scores were given for never/seldom being depressed/sad (55.3%) and never/seldom feeling lonely (71.8%). Approximately every second respondent (51.9%) had plans for the future. A clear decreasing trend was seen in all questions for respondents aged 75 in comparison to those aged 65: only ;;are you satisfied with life?'' showed no significant differences. Language/culture seems to clearly affect experiences of life orientation and well-being. Additionally, older people living alone reported significantly more negative response to all dimensions of life orientation. Positive life orientation is an important inner health resource for older. Accordingly, health promotion for older people should focus on facilitating the maintenance of important and close relationships and should include supporting older people's need to be needed.


A head-to-head comparison of the efficacy and safety of ixekizumab and adalimumab in biological-naïve patients with active psoriatic arthritis: 24-week results of a randomised, open-label, blinded-assessor trial.
To compare efficacy and safety of ixekizumab (IXE) to adalimumab (ADA) in biological disease-modifying antirheumatic drug-naïve patients with both active psoriatic arthritis (PsA) and skin disease and inadequate response to conventional synthetic disease-modifying antirheumatic drug (csDMARDs). Patients with active PsA were randomised (1:1) to approved dosing of IXE or ADA in an open-label, head-to-head, blinded assessor clinical trial. The primary objective was to evaluate whether IXE was superior to ADA at week 24 for simultaneous achievement of a ≥50% improvement from baseline in the American College of Rheumatology criteria (ACR50) and a 100% improvement from baseline in the Psoriasis Area and Severity Index (PASI100). Major secondary objectives, also at week 24, were to evaluate whether IXE was: (1) non-inferior to ADA for achievement of ACR50 and (2) superior to ADA for PASI100 response. Additional PsA, skin, treat-to-target and quality-of-life outcome measures were assessed at week 24. The primary efficacy endpoint was met (IXE: 36%, ADA: 28%; p=0.036). IXE was non-inferior for ACR50 response (IXE: 51%, ADA: 47%; treatment difference: 3.9%) and superior for PASI100 response (IXE: 60%, ADA: 47%; p=0.001). IXE had greater response versus ADA in additional PsA, skin, nail, treat-to-target and quality-of-life outcomes. Serious adverse events were reported in 8.5% (ADA) and 3.5% (IXE) of patients. IXE was superior to ADA in achievement of simultaneous improvement of joint and skin disease (ACR50 and PASI100) in patients with PsA and inadequate response to csDMARDs. Safety and tolerability for both biologicals were aligned with established safety profiles.


Interleukin-6 inhibition of hERG underlies risk for acquired long QT in cardiac and systemic inflammation.
Increased proinflammatory interleukin-6 (IL-6) levels are associated with acquired long QT-syndrome (LQTS) in patients with systemic inflammation, leading to higher risks for life-threatening polymorphic ventricular tachycardia such as Torsades de Pointes. However, the functional and molecular mechanisms of this association are not known. In most cases of acquired LQTS, the target ion channel is the human ether-á-go-go-related gene (hERG) encoding the rapid component of the delayed rectifier K current, IKr, which plays a critical role in cardiac repolarization. Here, we tested the hypothesis that IL-6 may cause QT prolongation by suppressing IKr. Electrophysiological and biochemical assays were used to assess the impact of IL-6 on the functional expression of IKr in HEK293 cells and adult guinea-pig ventricular myocytes (AGPVM). In HEK293 cells, IL-6 alone or in combination with the soluble IL-6 receptor (IL-6R), produced a significant depression of IKr peak and tail current densities. Block of IL-6R or Janus kinase (JAK) reversed the inhibitory effects of IL-6 on IKr. In AGPVM, IL-6 prolonged action potential duration (APD) which was further prolonged in the presence of IL-6R. Similar to heterologous cells, IL-6 reduced endogenous guinea pig ERG channel mRNA and protein expression. The data are first to demonstrate that IL-6 inhibition of IKr and the resulting prolongation of APD is mediated via IL-6R and JAK pathway activation and forms the basis for the observed clinical QT interval prolongation. These novel findings may guide the development of targeted anti-arrhythmic therapeutic interventions in patients with LQTS and inflammatory disorders.


Methyltransferase inhibitors restore SATB1 protective activity against cutaneous T cell lymphoma in mice.
Cutaneous T cell lymphoma (CTCL) has a poorly understood etiology and no known cure. Using conditional knockout mice, we found that ablation of the genomic organizer special AT-rich sequence-binding protein 1 (Satb1) caused malignant transformation of mature, skin-homing, Notch-activated CD4+ and CD8+ T cells into progressively fatal lymphoma. Mechanistically, Satb1 restrained Stat5 phosphorylation and the expression of skin-homing chemokine receptors in mature T cells. Notably, methyltransferase-dependent epigenetic repression of SATB1 was universally found in human Sézary syndrome, but not in other peripheral T cell malignancies. H3K27 and H3K9 trimethylation occluded the SATB1 promoter in Sézary cells, while inhibition of SUV39H1/2 methyltransferases (unlike EZH2 inhibition) restored protective SATB1 expression and selectively abrogated the growth of primary Sézary cells more effectively than romidepsin. Therefore, inhibition of methyltransferases that silence SATB1 could address an unmet need for patients with mycosis fungoides/Sézary syndrome, a set of incurable diseases.


Effect of a total work-site smoking ban on employee smoking and attitudes.
A "smoke-free" policy was adopted at the Duke University Medical Center but not at the adjacent University Campus. Three months after the smoking prohibition went into effect, a cross-sectional telephone survey was conducted, using randomly selected groups of 400 employees from each campus. Subjects were queried about current and previous smoking histories and their opinion of the smoking ban. As determined retrospectively from this survey, at the time of the announcement of the policy and 6 months before implementation, 23.6% of employees at the Medical Center were smokers, compared with 20.3% on the University Campus. Three months after implementation of the Medical Center smoking prohibition, smoking cessation rates were 12.6% at the Medical Center and 6.9% on the University Campus dating back 9 months to the time of policy announcement (P less than 0.10). Mean cigarette consumption during work hours declined over this same period from 8.1 +/- 6.8 (mean +/- SD) to 4.3 +/- 4.4 at the Medical Center but showed little change on the University Campus (9.3 +/- 7.5 v 8.7 +/- 8.0). Overall, 75.8% of subjects at the Medical Center "somewhat" or "strongly" agreed with the policy compared with 73.2% on the University Campus. A follow-up survey of the cohort of current or recent ex-smokers identified on the initial survey was conducted 6 months later. This survey revealed a smoking cessation rate of 22.5% at the Medical Center and 6.9% on the University Campus, dating back 15 months to the time of policy announcement (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


New Fathers' Perinatal Depression and Anxiety-Treatment Options: An Integrative Review.
More than 10% of fathers experience depression and anxiety during the perinatal period, but paternal perinatal depression (PPND) and anxiety have received less attention than maternal perinatal mental health problems. Few mainstream treatment options are available for men with PPND and anxiety. The aim of this literature review was to summarize the current understanding of PPND and the treatment programs specifically designed for fathers with perinatal depression. Eight electronic databases were searched using a predefined strategy, and reference lists were also hand searched. PPND and anxiety were identified to have a negative impact on family relationships, as well as the health of mothers and children. Evidence suggests a lack of support and tailored treatment options for men having trouble adjusting to the transition to fatherhood. Of the limited options available, cognitive behavioral therapy, group work, and blended delivery programs, including e-support approaches appear to be most effective in helping fathers with perinatal depression and anxiety. The review findings have important implications for the understanding of PPND and anxiety. Future research is needed to address the adoption of father-inclusive and father-specific models of care to encourage fathers' help-seeking behavior. Inclusion of male-specific requirements into support and treatment options can improve the ability of services to engage new fathers. Psychotherapeutic intervention could assist to address the cognitive differences and dissonance for men adjusting to the role of father, including male identity and role expectations.


Karyotype differentiation in Chromaphyosemion killifishes (Cyprinodontiformes, Nothobranchiidae). I: Chromosome banding patterns of C. alpha, C. kouamense and C. lugens.
The karyotypes of three recently described species of Chromaphyosemion, namely C. lugens, C. alpha and C. kouamense, were analysed using conventional Giemsa staining, C-banding and sequential banding (fluorescence banding with 4', 6-diamidino-2-phenylindole (DAPI) and Chromomycin A(3) (CMA(3)), C-banding, AgNO(3)-staining). Diploid chromosome numbers ranged from 2n = 36 in C. lugens to 2n = 38 in C. alpha and C. kouamense. The karyotype of C. lugens consisted of one pair of metacentric (m) and 17 pairs of telocentric (t) chromosomes, that of C. alpha was composed of one pair of submetacentric (sm), six pairs of subtelocentric (st) and 12 pairs of t chromosomes, and that of C. kouamense comprised five pairs of st and 14 pairs of t chromosomes. In addition to the differences in karyotype structures and/or chromosome numbers, the karyotypes of the examined species differed with respect to NOR phenotype and distribution and base composition of heterochromatin. No heteromorphic sex chromosomes were detected in any of the species. Our findings provide cytotaxonomic evidence for the species distinctness of C. alpha, C. kouamense and C. lugens whose descriptions were based primarily on external morphology.


Irrational antibiotic usage in boarding secondary school settings in Dar es Salaam.
To assess the prevalence of antibiotic misuse among boarding secondary school students in Dares Salaam. This descriptive cross-sectional study was conducted over 3-month period amongst boarding secondary school students. A randomized sample of students was recruited from at least 3 secondary schools from each of the three municipalities of Dar es Salaam region. Questionnaire with both closed-and open-ended questions was used for data collection in which socio-demographic characteristics, knowledge of ATBs, sources of prescription, procurement of ATBs, infections for which ATBs were used, and antibiotic therapy compliance were assessed. The collected data were analyzed using SPSS/PC+ version 15.0 computer package. A total of 424 randomly selected students with a mean age of 20 years were interviewed, of those 150 were females and 274 were males. Majority of the respondents (74%) have had heard of ATBs, however only 105 (25%) respondent described them correctly. Prevalence of antibiotic usage among the students was high (69%). Of 293 students who have had used ATBs, 73% of them had procured the drugs on prescription; 170 (58%) respondents used the ATBs for non-microbial infections/diseases, only 183 (62%) were compliant to antibiotic therapies (ABT), while 226 (77%) admitted to have had shared ATBs with friends/relatives whenever deemed necessary. Of those who had procured ATBs without prescriptions, 66% of them obtained the drugs from pharmacies. The study revealed inadequate knowledge on ATBs among the respondents and that pharmacies played a major role on easy availability of ATBs as over-the-counter (OTC) drugs attributing to high rate of irrational use of ATBs. We recommended that health care providers should succinctly explain to patients the importance of ABT compliance and adverse effects of non-compliance. Regulatory authorities should re-enforce the laws on sales of ATBs by prohibiting their availability as OTC drugs.


Platelet-activating factor (PAF) stimulates the production of PAF acetylhydrolase by the human hepatoma cell line, HepG2.
The human hepatoma cell line, HepG2, secreted an activity that degrades platelet-activating factor (PAF) by the hydrolysis of the sn-2 acetyl group. This activity was Ca++ independent, inhibited by diisopropylfluorophosphate but not by p-bromophenacyl bromide, and resistant to treatment with trypsin or pronase. Separation of HepG2-conditioned medium by gel filtration disclosed that the activity was associated with lipoproteins. An antiserum against PAF acetylhydrolase immunoprecipitated this activity. It was not recognized by an antibody against lecithin:cholesterol acyltransferase (LCAT), which also is secreted by HepG2 cells. Therefore the phospholipase A2 activity of LCAT was excluded as a source of the observed activity. PAF added to the culture medium stimulated the secretion of the PAF-degrading activity by HepG2 cells, while lyso-PAF was inactive. Maximal stimulation was observed with 5 ng/ml PAF, which induced a fivefold increase. The presence of 5 ng/ml PAF, enhanced the secretion of [35S]methionine-labeled PAF acetylhydrolase and cycloheximide inhibited both the basal and PAF-stimulated secretion of the labeled enzyme. We conclude that HepG2 cells produce PAF acetylhydrolase. The liver may be a major source of plasma PAF acetylhydrolase, and PAF may induce the production of its inactivating enzyme by the liver.


Gardening is beneficial for adult mental health: Scottish Health Survey, 2012-2013.
Gardening has been reported as being beneficial for mental well-being for vulnerable populations since 2000. However, little is known concerning its role in the general population. Therefore, the aim of the present study was to investigate the relationship of gardening and mental health in adults in a countrywide and population-based setting. Data was retrieved from and analysed in the Scottish Health Survey, 2012-2013. Information on demographics, lifestyle factors, gardening engagement, and adult mental health by General Health Questionnaire was obtained by household interview. Statistical analyses including chi-square test, t-test and survey-weighted logistic and multi-nominal regression modelling were performed. Of 9709 Scottish adults aged 16-99, 5 531 (57.0%) people did not do any gardening or building work in the last four weeks. A total of 888 (9.2%) people reported poor self-rated health. Gardening was associated with adult mental health in people both with or without heart conditions including ability to concentrate, feeling playing a useful part in things, feeling capable of making decisions, thinking of self as worthless, feeling reasonably happy, etc. General adults with or without heart conditions could benefit from engaging with gardening or building work. Future public health programmes promoting such activity should be encouraged in order to optimise adult mental health.


Single Port Laparoscopic Total and Subtotal Colectomies for Inflammatory Bowel Disease in a District General Hospital.
Background: Single incision laparoscopic surgery (SILS) is expanding, enhancing the advantages of multi-port laparoscopic surgery (MLS). Limited literature exists regarding SILS total/subtotal colectomies for inflammatory bowel disease (IBD). Aim of the study was to present the initial experience with this type of approach in a district general hospital and extrapolate its feasibility and safety in this specific context based on gold standard outcomes reported in literature. Materials and Methods: Preoperative parameters, operative details and surgical outcomes of consecutive patients who underwent colonic SILS for IBD in a 5-year period were reviewed retrospectively. Median length of follow-up was 26 months. Results: Fourteen patients underwent SILS subtotal/total colectomy. Median body mass index was 25 (18.1-35). Two patients had previous abdominal surgeries. Median operating time was 202.5 minutes. Two cases were converted to open. Median length of stay was 5 days. Three patients presented complications. Three patients developed parastomal hernias (21.4%). Five out of 12 patients with ulcerative colitis declined further surgery, 3 are awaiting laparoscopic/SILS pouch formation, 1 underwent SILS pouch formation, 1 SILS ileo-rectal anastomosis and 1 patient had SILS completion proctectomy. One patient was not followed up. Conclusions: Despite literature data heterogeneity, these results provide support to the feasibility and applicability of SILS in the subgroup of patients who undergo subtotal/total colectomies for IBD, offering the option for subsequent SILS completion or restorative procedures. Further studies are required to explore the benefit of SILS over MLS (including cosmesis and quality of life) and non-inferiority of SILS regarding the parastomal hernia issue and the operative duration.


Caregiving for the terminally ill: at what cost?
This literature review exposes the nature and extent of physical and psychosocial morbidity and economic disadvantage, home palliative caregivers suffer as a direct result of their caregiving role. Research has demonstrated that caregivers providing support to individuals receiving palliative care report unmet needs for information, communication, service provision and support from health and community services. Three sets of challenges are highlighted in this literature review which help explain why the needs of home palliative caregivers are largely unmet: (i) barriers to seeking help; (ii) a dearth of research-based interventions focused on reducing the negative aspects of caregiving; and (iii) a number of impediments to effective policy and service development for family caregivers. Furthermore, invited submissions from caregivers echoed and confirmed the issues reported in the literature. Recommendations for enhancing caregiver support are outlined.


[Clinical characteristics of pediatric patients with chronic constipation according to age group].
There was no study evaluating clinical characteristics of constipation according pediatric age groups. To evaluate the clinical characteristics of patients with chronic constipation according to age group. This retrospective study evaluated the demographic data and clinical characteristics of pediatric patients with chronic constipation who had been admitted to an outpatient clinic between May 1995 and December 2000. Data was analyzed according to the followings age groups: infants, pre-school, school age and adolescent. Five hundred and sixty one patients were included in the study. The percentage of patients according to age groups were: infants (19.1%), pre-school (42.9%), school age (26.9%), and adolescents (11.0%). There was no statistical gender difference among the four age groups. Less than three bowel movements per week were observed more frequently in pre-school (65.8%) and school age (59.6%) than in infants (52.4%) and adolescents (43.1%). Fecal escape ("soiling") was found in 75.6% of the pre-school patients aged more than 48 months, in 68.2% of the school age and in 76.7% of the adolescents. Soiling was more frequent in boys (80.7%) than in girls (50.0%) only in school age children. Retentive posture (67.6%, 40.7%, 27.2%), fear for defecation (70.2%, 44.2%, 29.7%) and abdominal pain (80.8%, 69.6%, 73.6%) were observed, respectively, more frequently in pre-school than in school age and adolescents. There were differences in clinical characteristics according to age group. However, prolonged duration of constipation and an elevated number of complications were observed in all age groups, especially fecal soiling and abdominal pain.


What sleeping babies hear: a functional MRI study of interparental conflict and infants' emotion processing.
Experiences of adversity in the early years of life alter the developing brain. However, evidence documenting this relationship often focuses on severe stressors and relies on peripheral measures of neurobiological functioning during infancy. In the present study, we employed functional MRI during natural sleep to examine associations between a more moderate environmental stressor (nonphysical interparental conflict) and 6- to 12-month-old infants' neural processing of emotional tone of voice. The primary question was whether interparental conflict experienced by infants is associated with neural responses to emotional tone of voice, particularly very angry speech. Results indicated that maternal report of higher interparental conflict was associated with infants' greater neural responses to very angry relative to neutral speech across several brain regions implicated in emotion and stress reactivity and regulation (including rostral anterior cingulate cortex, caudate, thalamus, and hypothalamus). These findings suggest that even moderate environmental stress may be associated with brain functioning during infancy.


Immunogenicity and protective efficacy of mycobacterial DNA vaccines incorporating plasmid-encoded cytokines against Mycobacterium bovis.
DNA-based vaccines, alone or in combination with other sub-unit vaccination regimes, represent an alternative to live mycobacterial vaccines for protective immunization against tuberculosis. Here, we have used a murine immunization or Mycobacterium bovis aerosol challenge model to assess the immunogenicity and protective efficacy of mycobacterial DNA vaccines. Mice that received immunization with DNA constructs encoding M. bovis antigen 85A (Ag85-A) and arget(ESAT-6) produced measurable interferon-gamma (IFN-gamma) responses to CD4(+) T-cell epitope-peptide recall antigens in vitro. The magnitude of these responses was enhanced by co-delivery of a construct encoding murine cytokines (macrophage inhibitory protein (MIP)-1 alpha or interleukin(IL)-7), although they did not the match responses observed in mice that received Bacille Calmette-Guerin(BCG) immunisation. In contrast, DNA priming followed by boosting with modified vaccinia Ankara (MVA) vaccine (expressing M. tuberculosis Ag85-A) invoked higher IFN-gamma levels, with the most immunogenic regime of Ag85 or ESAT or IL-7 prime followed by MVA boost being of commensurate immunogenicity to BCG. Despite this, neither DNA alone nor DNA-prime or MVA boost regimes conferred measurable protection against aerosol challenge with virulent M. bovis. These data highlight both the promise and the shortcomings of new generation subunit tuberculosis vaccines, with particular emphasis on their potential as vaccines against M. bovis.


Caffe con Troll: Shallow Ideas to Speed Up Deep Learning.
We present Caffe con Troll (CcT), a fully compatible end-to-end version of the popular framework Caffe with rebuilt internals. We built CcT to examine the performance characteristics of training and deploying general-purpose convolutional neural networks across different hardware architectures. We find that, by employing standard batching optimizations for CPU training, we achieve a 4.5× throughput improvement over Caffe on popular networks like CaffeNet. Moreover, with these improvements, the end-to-end training time for CNNs is directly proportional to the FLOPS delivered by the CPU, which enables us to efficiently train hybrid CPU-GPU systems for CNNs.


Fluoride inhibition of the hydro-osmotic response of the toad urinary bladder to antidiuretic hormone.
The hydro-osmotic response of the toad bladder to antidiuretic hormone and cyclic AMP was inhibited by the methoxyflurane metabolite, fluoride. The osmotic transfer of water in the absence of hormone was unaffected by fluoride as was the hydroosmotic response due to hypertonicity of the serosal bathing media. Osmotic water movements across N-ethylmaleimide-"fixed" vasopressin or cyclic AMP-stimulated bladders were likewise unchanged by fluoride, suggesting that fluoride is exerting an action subsequent to the endogenous formation of cyclic AMP but before the final effector mechanism. Fluoride increased intracellular cyclic AMP concentrations even in the presence of added hormone. Fluoride suppressed calmodulin activity and prevented its activation of phosphodiesterase. Fluoride had no effect on oxygen consumption of toad urinary bladder cells but reduced lactate formation and anerobic metabolism. This decrease in the glycolytic energy source did not contribute to the inhibition of the hormonal response since 2-deoxyglucose was without effect on hormonal mediated osmotic-water flow. It is postulated that the fluoride-induced polyuria after methoxyflurane anesthesia may be due in part to the ability of fluoride to interfere with calcium and calmodulin-initiated processes (other than phosphodiesterase activity) that may occur in the stimulus-reabsorption coupling response of antidiuretic hormone.


Loss of hsp70.1 Decreases Functional Motor Recovery after Spinal Cord Injury in Mice.
Heat shock proteins (HSPs) are specifically induced by various forms of stress. Hsp70.1, a member of the hsp70 family is known to play an important role in cytoprotection from stressful insults. However, the functional role of Hsp70 in motor function after spinal cord injury (SCI) is still unclear. To study the role of hsp70.1 in motor recovery following SCI, we assessed locomotor function in hsp70.1 knockout (KO) mice and their wild-type (WT) mice via the Basso, Beattie and Bresnahan (BBB) locomotor rating scale, before and after spinal hemisection at T13 level. We also examined lesion size in the spinal cord using Luxol fast blue/cresyl violet staining. One day after injury, KO and WT mice showed no significant difference in the motor function due to complete paralysis following spinal hemisection. However, when it compared to WT mice, KO mice had significantly delayed and decreased functional outcomes from 4 days up to 21 days after SCI. KO mice also showed significantly greater lesion size in the spinal cord than WT mice showed at 21 days after spinal hemisection. These results suggest that Hsp70 has a protective effect against traumatic SCI and the manipulation of the hsp70.1 gene may help improve the recovery of motor function, thereby enhancing neuroprotection after SCI.


Coumarins as antioxidants.
Coumarins, a well-known class of naturally occurring compounds, display a remarkable array of biochemical and pharmacological actions, some of which suggest that certain members of this group of compounds may significantly affect the function of various mammalian cellular systems. The development of coumarins as antioxidant agents has attracted much attention in recent years. Coumarins afford an opportunity for the discovery of new antioxidants with truly novel mechanisms of action. This review updates and expands the 2006 review by the same author. The review considers and incorporates the most recently published literature on coumarins as related to their antioxidant properties. A lot of coumarins have been identified from natural sources, especially green plants. These natural compounds have served as valuable leads for further design and synthesis of more active analogues. Beyond doubt, a deep understanding of the mechanisms of existing synthetic and natural coumarins will build the basis for the rational design.


Scolders, carers or friends: South African midwives' contrasting styles of communication when discussing smoking cessation with pregnant women.
to investigate how midwives are currently communicating with women about smoking during pregnancy with a view to involving them in a smoking cessation intervention in antenatal clinics. a qualitative study using individual, in-depth interviews for data collection. 24 nurses providing antenatal care to pregnant smokers attending public sector clinics in five major cities in South Africa. three archetypes of midwives, characterised by different styles of communication and approaches to smoking cessation, emerged from the analysis of the interview data. These were described as the 'Angry Scolders', the 'Benign Carers' and the 'Enthusiastic Friends'. The first type conformed to the traditional, authoritarian style of communication, where the midwife assumed a dominant, expert role. When women failed to comply with their advice, these midwives typically became angry and confrontational. The second type of midwife used a paternalistic communication style and emphasised the role of education in changing behaviour. However, these midwives had little confidence that they could influence women to quit. The third type embraced a patient-centred approach, consciously encouraging more interaction with their patients and attempting to understand change from their point of view. These midwives were optimistic of women's capacity to change and more satisfied with their current health education efforts than the first two types. The Benign Carers and Enthusiastic Friends were more open to participation in the potential intervention than the Angry Scolders. the prevailing traditional, authoritarian style of communication is inappropriate for smoking cessation education and counselling as it provokes resistance and avoidance on the part of pregnant smokers. The paternalistic approach appears to be largely ineffectual, whereas the patient-centred approach elicits the most positive response from pregnant women and enhances the possibility of a trusting and cooperative relationship with the midwife. Midwives using this style are more open to fulfilling their role in smoking cessation. smoking cessation interventions need to attend to not only what midwives say to pregnant women about smoking, but also how they communicate about the issue. The use of a patient-centred approach, such as brief motivational interviewing, is recommended as a means of improving counselling outcomes among pregnant smokers.


A combination of physical activity and computerized brain training improves verbal memory and increases cerebral glucose metabolism in the elderly.
Physical exercise interventions and cognitive training programs have individually been reported to improve cognition in the healthy elderly population; however, the clinical significance of using a combined approach is currently lacking. This study evaluated whether physical activity (PA), computerized cognitive training and/or a combination of both could improve cognition. In this nonrandomized study, 224 healthy community-dwelling older adults (60-85 years) were assigned to 16 weeks home-based PA (n=64), computerized cognitive stimulation (n=62), a combination of both (combined, n=51) or a control group (n=47). Cognition was assessed using the Rey Auditory Verbal Learning Test, Controlled Oral Word Association Test and the CogState computerized battery at baseline, 8 and 16 weeks post intervention. Physical fitness assessments were performed at all time points. A subset (total n=45) of participants underwent [(18)F] fluorodeoxyglucose positron emission tomography scans at 16 weeks (post-intervention). One hundred and ninety-one participants completed the study and the data of 172 participants were included in the final analysis. Compared with the control group, the combined group showed improved verbal episodic memory and significantly higher brain glucose metabolism in the left sensorimotor cortex after controlling for age, sex, premorbid IQ, apolipoprotein E (APOE) status and history of head injury. The higher cerebral glucose metabolism in this brain region was positively associated with improved verbal memory seen in the combined group only. Our study provides evidence that a specific combination of physical and mental exercises for 16 weeks can improve cognition and increase cerebral glucose metabolism in cognitively intact healthy older adults.


Navigating 'ethics in practice': An ethnographic case study with young women living with HIV in Zambia.
While 'procedural ethics' provides essential frameworks for governing global health research, reflecting on 'ethics in practice' offers important insights into addressing ethically important moments that arise in everyday research. Particularly for ethnographic research, renowned for it's fluid and spontaneous nature, engaging with 'ethics in practice' has the potential to enhance research practice within global health. We provide a case study for such reflexivity, exploring 'ethics in practice' of ethnographic research with middle-income young women living with HIV in Lusaka, Zambia. We explore the ethical issues arising from the layered interaction of the population (young women), the disease under investigation (HIV), the method of study (ethnographic), and the setting (Zambia, a lower middle income country). We describe how we navigated five key practical ethical tensions that arose, namely the psycho-emotional benefits of the research, the negotiated researcher-participant relationship, protecting participants' HIV status, confidentiality and data ownership, and researcher obligations after the end of the research. We exemplify reflexive engagement with 'ethics in practice' and suggest that engaging with ethics in this way can make important contributions towards developing more adequate ethical guidelines and research practice in global public health.


Polyol formation and NADPH-dependent reductases in dog retinal capillary pericytes and endothelial cells.
Dogs fed a diet containing 30% galactose experience retinal vascular changes similar to those in human diabetic retinopathy, with selective pericyte loss as an initial lesion. In the present study the relationship among reduced nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reductases, polyol formation, and flux through the polyol pathway in cultured dog retinal capillary cells were investigated. Pericytes and endothelial cells were cultured from retina of beagle dogs. NADPH-dependent reductases were characterized by chromatofocusing after gel filtration. Sugars in cultured cells were analyzed by gas chromatography, and flux through the polyol pathway was investigated by 19F nuclear magnetic resonance (NMR) with 3-fluoro-3-deoxy-D-glucose (3FG) as a substrate. The presence of aldose reductase and sorbitol dehydrogenase in these cells was examined by northern blot analysis. Two distinct peaks corresponding to aldose reductase and aldehyde reductase, the latter being dominant, were observed in pericytes by chromatofocusing. Culture in medium containing either 10 mM D-galactose or 30 mM D-glucose resulted in the accumulation of sugar alcohol in pericytes that was markedly reduced by aldose reductase inhibitors. 19F NMR spectra obtained from pericytes cultured for 5 days in medium containing 2 mM 3FG displayed the marked accumulation of 3-fluoro-deoxysorbitol but not 3-fluoro-deoxyfructose. No 3FG metabolism was observed in similarly cultured endothelial cells. With northern blot analysis, aldose reductase was detected in pericytes but not in endothelial cells. Sorbitol dehydrogenase was below the detectable limit in pericytes and endothelial cells. Aldose, aldehyde, and glyceraldehyde reductases are present in dog retinal capillary pericytes, with aldehyde reductase being the major reductase present. Polyol accumulation easily occurs in pericytes but not in endothelial cells.


Weight discrimination and unhealthy eating-related behaviors.
Individuals with obesity often experience unfair treatment because of their body weight. Such experiences are associated with binge eating, but less is known about its association with other eating-related behaviors and whether these relations are specific to discrimination based on weight or extend to other attributions for discrimination. The present research uses a large national sample (N = 5129) to examine whether weight discrimination is associated with diet and meal rhythmicity, in addition to overeating, and whether these associations generalize to nine other attributions for discrimination. We found that in addition to overeating, weight discrimination was associated with more frequent consumption of convenience foods and less regular meal timing. These associations were generally similar across sex, age, and race. Discrimination based on ancestry, gender, age, religion, and physical disability were also associated with overeating, which suggests that overeating may be a general coping response to discrimination. Unfair treatment because of body weight is associated with unhealthy eating-related behaviors, which may be one pathway through which weight discrimination increases risk for weight gain and obesity.


Burden of stroke in Bangladesh.
Stroke is the third leading cause of death in Bangladesh. The World Health Organization ranks Bangladesh's mortality rate due to stroke as number 84 in the world. The reported prevalence of stroke in Bangladesh is 0.3%, although no data on stroke incidence have been recorded. Hospital-based studies conducted in past decades have indicated that hypertension is the main cause of ischaemic and haemorrhagic stroke in Bangladesh. The high number of disability-adjusted life-years lost due to stroke (485 per 10,000 people) show that stroke severely impacts Bangladesh's economy. Although two non-governmental organizations, BRAC and the Centre for the Rehabilitation of the Paralysed, are actively involved in primary stroke prevention strategies, the Bangladeshi government needs to emphasize healthcare development to cope with the increasing population density and to reduce stroke occurrence.


Differential outcome of pure manic, mixed/cycling, and pure depressive episodes in patients with bipolar illness.
We found significant differences in time to recovery and rates of chronicity in 155 patients with bipolar illness when the episodes were subtyped into those with manic symptoms alone (pure manic), depressive symptoms alone (pure depressed), or symptoms of depression and mania (mixed or cycling) up to the time of entry into a clinical research study. Most of the patients in all three groups who did not recover received levels of somatotherapy that were generally consistent with current recommendations for intensity of treatment appropriate to each condition. Based on a median follow-up of 18 months, the life-table estimate of the probability of remaining ill for at least one year was 7% for the pure manic patients compared with 32% in patients who entered the study with episodes that were mixed or cycling. Purely depressed patients had a 22% probability of remaining ill, approximating rates found in patients without bipolar illness who have episodes of depression. Different clinical variables were found to predict time to recovery in each of these groups. We propose that this subtyping of episodes may be a clinically useful part of the classification of bipolar disorders.


A meta-analysis of cognitive impairment following adult cancer chemotherapy.
Chemotherapy-induced cognitive impairments are reported by many cancer survivors. Research to date has not provided a clear description of their nature, extent, mechanisms, and duration. To investigate the impairments and factors that could influence their identification and severity, the present meta-analysis brings together research on this topic in adult cancer patients. Our random-model meta-analysis includes 44 studies investigating the cognitive performance of adults treated with chemotherapy for non-central nervous system malignancies, primarily breast and testicular cancer. We conducted several subgroup analyses to identify the level of cognitive impairments in longitudinal and cross-sectional studies. We also pursued several multilevel model regressions to investigate the impact of methodological (study quality) and clinical moderators (diagnosis, age, time since treatment) on the observed effect sizes. Cognitive impairments were found in cross-sectional studies in immediate free recall, delayed memory, verbal memory, delayed recognition memory, selective attention, and attention capacity. Surprisingly, prior to chemotherapy, patients performed better than matched controls. In longitudinal studies, patients' performance increased from baseline to follow-up, an effect that was stronger in patients than controls. None of the chosen moderators influenced the magnitude of estimated summary effect sizes. The likelihood to identify impairments rests on the type of design employed, as memory and attention impairments are only detected in cross-sectional studies. We discuss the lack of significant impact of moderators on the effect sizes despite the heterogeneity of results, while providing recommendations toward decreasing the heterogeneity in future studies.


Current and future management of bilateral loss of vestibular sensation - an update on the Johns Hopkins Multichannel Vestibular Prosthesis Project.
Bilateral loss of vestibular sensation can disable individuals whose vestibular hair cells are injured by ototoxic medications, infection, Ménière's disease or other insults to the labyrinth including surgical trauma during cochlear implantation. Without input to vestibulo-ocular and vestibulo-spinal reflexes that normally stabilize the eyes and body, affected patients suffer blurred vision during head movement, postural instability, and chronic disequilibrium. While individuals with some residual sensation often compensate for their loss through rehabilitation exercises, those who fail to do so are left with no adequate treatment options. An implantable neuroelectronic vestibular prosthesis that emulates the normal labyrinth by sensing head movement and modulating activity on appropriate branches of the vestibular nerve could significantly improve quality of life for these otherwise chronically dizzy patients. This brief review describes the impact and current management of bilateral loss of vestibular sensation, animal studies supporting the feasibility of prosthetic vestibular stimulation, and a vestibular prosthesis designed to restore sensation of head rotation in all directions. Similar to a cochlear implant in concept and size, the Johns Hopkins Multichannel Vestibular Prosthesis (MVP) includes miniature gyroscopes to sense head rotation, a microcontroller to process inputs and control stimulus timing, and current sources switched between pairs of electrodes implanted within the vestibular labyrinth. In rodents and rhesus monkeys rendered bilaterally vestibulardeficient via treatment with gentamicin and/or plugging of semicircular canals, the MVP partially restores the vestibulo-ocular reflex for head rotations about any axis of rotation in 3-dimensional space. Our efforts now focus on addressing issues prerequisite to human implantation, including refinement of electrode designs and surgical technique to enhance stimulus selectivity and preserve cochlear function, optimization of stimulus protocols, and reduction of device size and power consumption.


Intensity-modulated fractionated stereotactic radiotherapy with reduced margin for high grade gliomas: dosimetric analysis of sparing optic nerve & chiasm.
The aim of this study was to evaluate the potential advantage of intensity modulation fractionated stereotactic radiotherapy (IM-FSRT) for sparing the optic nerve and chiasm in the management of anaplastic astrocytoma (AA) or glioblastoma (GBM). Clinical data of 16 patients with high grade gliomas were retrospectively reviewed. All patients had tumors close to the optic chiam that prohibited the full dose radiation therapy. The patients were treated with either IM-FSRT in conjunction with external beam radiation or IM-FSRT only to a total dose of 60 Gy. The final dosimetric analysis was performed. The estimated average maximum optic nerve and chiasm dose 48.6±5.0 Gy (range, 34.2-55.9). Conformity index reached as high as 0.98, and inhomogeneity coefficient was low enough to deliver the prescribed radiation dose. IM-FSRT showed excellent dose conformality to the target volume with superior normal tissue sparing effect. High precision therapy such as IM-FSRT could be a reliable treatment technique for the patients with AA or GBM for a maximal sparing of optic apparatus.


Edin Expression in the Fat Body Is Required in the Defense Against Parasitic Wasps in Drosophila melanogaster.
The cellular immune response against parasitoid wasps in Drosophila involves the activation, mobilization, proliferation and differentiation of different blood cell types. Here, we have assessed the role of Edin (elevated during infection) in the immune response against the parasitoid wasp Leptopilina boulardi in Drosophila melanogaster larvae. The expression of edin was induced within hours after a wasp infection in larval fat bodies. Using tissue-specific RNAi, we show that Edin is an important determinant of the encapsulation response. Although edin expression in the fat body was required for the larvae to mount a normal encapsulation response, it was dispensable in hemocytes. Edin expression in the fat body was not required for lamellocyte differentiation, but it was needed for the increase in plasmatocyte numbers and for the release of sessile hemocytes into the hemolymph. We conclude that edin expression in the fat body affects the outcome of a wasp infection by regulating the increase of plasmatocyte numbers and the mobilization of sessile hemocytes in Drosophila larvae.


Integrated electromyography of the masseter on incremental opening and closing with audio biofeedback: a study on mandibular posture.
The purpose of this study was to test the hypothesis of a minimum electromyographic (EMG) rest position based on masseter surface EMG recordings of incremental opening and closing of the mandible with simultaneous audio EMG biofeedback. Nineteen alert subjects in an upright seated position opened and closed the mandible in 1-mm increments 20 mm interincisally. An electronic recording device allowed each subject to maintain the vertical dimension of each increment while simultaneously reducing right masseteric muscle activity to the minimum possible level using audio EMG biofeedback. Integrated EMG masseteric activity was recorded at each static opening and closing increment. A mean plateau of integrated EMG output for all subjects with no minimum EMG point or circumscribed minimum EMG range for any of the nineteen subjects was shown. Analysis of variance for repeated measures showed no difference in opening and closing EMG levels (P = 0.27) and no interaction between opening, closing, and change in vertical dimension (P < 0.0001). These results, with those of other studies, raise questions regarding the validity of the concept of a unique physiologic rest position of the mandible with the masseter or associated muscles at minimum muscle activity. The idea of overlapping postural ranges appears to be more appropriate.


Enantiomer/enantiomer interaction of (S)- and (R)-propafenone for cytochrome P450IID6-catalyzed 5-hydroxylation: in vitro evaluation of the mechanism.
Many drugs are used as racemates, and the enantiomers may differ in terms of pharmacological properties and disposition. Stereoselective disposition of the enantiomers can arise from metabolism of the enantiomers via different routes catalyzed by different enzymes. In contrast, the enantiomers may be metabolized by the same enzyme at different rates. In the latter case, the enantiomers can compete for this metabolic step, giving rise to the possibility of an enantiomer/enantiomer interaction. We have chosen the antiarrhythmic propafenone, for which in vivo data indicated an interaction between (S)- and (R)-propafenone, as a model substance to study the mechanism underlying that interaction in human liver microsomes. We used the cytochrome P450IID6-mediated 5-hydroxylation of propafenone as a model pathway, because this metabolic step constitutes the major route of biotransformation of propafenone. The Michaelis-Menten kinetics for 5-hydroxylation were determined after incubation of (R)- and (S)-propafenone and a pseudoracemate consisting of (S)-[2H4]propafenone and (R)-propafenone. Inhibition experiments were performed using (S)-[2H4]propafenone as an inhibitor of the 5-hydroxylation of (R)-propafenone, and vice versa. The kinetic model of mixed alternative substrates was used to simulate inhibition experiments. Experimental data were compared with those predicted by this model. We observed a substantial stereoselectivity after incubation of the individual enantiomers [(S)-propafenone: Vmax, 10.2 pmol/micrograms/hr, and Km, 5.3 microM; (R)-propafenone: Vmax, 5.5 pmol/micrograms/hr, and Km, 3.0 microM]. In contrast, no substrate stereoselectivity was observed after incubation of the pseudoracemate [3.1 pmol/micrograms/hr for (S)-[2H4]propafenone and 3.3 pmol/micrograms/hr for (R)-propafenone]. Application of the model revealed Ki values of 2.9 and 5.2 microM for the inhibition of 5-hydroxylation of (S)-[2H4]-propafenone by (R)-propafenone and for inhibition of 5-hydroxylation of (R)-propafenone by (S)-[2H4]-propafenone, respectively. The predicted and the experimental data were in good agreement, and both indicated the mode of inhibition to be competitive. In conclusion, the enantiomers of propafenone interact with respect to 5-hydroxylation, with (R)-propafenone being a more potent inhibitor than the S-enantiomer with respect to cytochrome P450IID6-mediated 5-hydroxylation. Because beta-blocking properties of propafenone reside in the S-enantiomer, inhibition of metabolism of this enantiomer by (R)-propafenone may have therapeutic consequences.


Grouping puts figure-ground assignment in context by constraining propagation of edge assignment.
Figure-ground organization involves the assignment of edges to a figural shape on one or the other side of each dividing edge. Established visual cues for edge assignment primarily concern relatively local rather than contextual factors. In the present article, we show that an assignment for a locally unbiased edge can be affected by an assignment of a remote contextual edge that has its own locally biased assignment. We find that such propagation of edge assignment from the biased remote context occurs only when the biased and unbiased edges are grouped. This new principle, whereby grouping constrains the propagation of figural edge assignment, emerges from both subjective reports and an objective short-term edge-matching task. It generalizes from moving displays involving grouping by common fate and collinearity, to static displays with grouping by similarity of edge-contrast polarity, or apparent occlusion. Our results identify a new contextual influence on edge assignment. They also identify a new mechanistic relation between grouping and figure-ground processes, whereby grouping between remote elements can constrain the propagation of edge assignment between those elements. Supplemental materials for this article may be downloaded from http://app.psychonomic-journals.org/content/supplemental.


New insights into the regulation of glucagon secretion by glucagon-like peptide-1.
Glucagon-like peptide-1 (GLP-1) is a potent incretin hormone currently under investigation for use as a novel therapeutic agent in the treatment of type 2 diabetes. One of several therapeutically important biological actions of GLP-1 in type 2 diabetic subjects is ability to induce strong suppression of glucagon secretion. The glucagonostatic action of GLP-1 results from its interaction with a specific G-protein coupled receptor resulting in the activation of adenylate cyclase and an increase in cAMP generation. In the pancreatic alpha-cell, cAMP, via activation of protein kinase A, interacts with a plethora of signal transduction processes including ion-channel activity and exocytosis of the glucagon-containing granules. In this short review, we will focus on recent advances in our understanding on the cellular mechanisms proposed to underlie the glucagonotropic action of GLP-1 and attempt to incorporate this knowledge into a working model for the control of glucagon secretion. Studies on the effects of GLP-1 on glucagon secretion are relevant to the pathogenesis of type 2 diabetes due to the likely contribution of hyperglucagonemia to impaired glucose tolerance in type 2 diabetes.


Encoding Growth Factor Identity in the Temporal Dynamics of FOXO3 under the Combinatorial Control of ERK and AKT Kinases.
Extracellular growth factors signal to transcription factors via a limited number of cytoplasmic kinase cascades. It remains unclear how such cascades encode ligand identities and concentrations. In this paper, we use live-cell imaging and statistical modeling to study FOXO3, a transcription factor regulating diverse aspects of cellular physiology that is under combinatorial control. We show that FOXO3 nuclear-to-cytosolic translocation has two temporally distinct phases varying in magnitude with growth factor identity and cell type. These phases comprise synchronous translocation soon after ligand addition followed by an extended back-and-forth shuttling; this shuttling is pulsatile and does not have a characteristic frequency, unlike a simple oscillator. Early and late dynamics are differentially regulated by Akt and ERK and have low mutual information, potentially allowing the two phases to encode different information. In cancer cells in which ERK and Akt are dysregulated by oncogenic mutation, the diversity of states is lower.


Time-resolved diffuse optical tomography system using an accelerated inverse problem solver.
A computationally efficient time-resolved diffuse optical tomography (TR-DOT) prototype was demonstrated using an accelerated inverse problem solver to reconstruct high quality 3D images of highly scattering media such as tissues. The inverse problem solver utilizes seven well-defined points on each experimentally recorded histogram of the distribution time-of-flight (DToF). In this work, the accuracy of the recovered optical properties, and the computational load and time of TR-DOT prototype were investigated using cylindrical turbid phantoms. These phantoms were measured using transmittance geometry under different conditions in multiple experiments to evaluate the performance of this prototype. Overall, the results of evaluation are important in the realization of a real-time and highly accurate TR-DOT system for diffuse optical imaging applications.


Distinct mesenchymal alterations in N-cadherin and E-cadherin positive primary renal epithelial cells.
Renal tubular epithelial cells of proximal and distal origin differ markedly in their physiological functions. Therefore, we hypothesized that they also differ in their capacity to undergo epithelial to mesenchymal alterations. We used cultures of freshly isolated primary human tubular cells. To distinguish cells of different tubular origin we took advantage of the fact that human proximal epithelial cells uniquely express N-cadherin instead of E-cadherin as major cell-cell adhesion molecule. To provoke mesenchymal alteration we treated these cocultures with TGF-β for up to 6 days. Within this time period, the morphology of distal tubular cells was barely altered. In contrast to tubular cell lines, E-cadherin was not down-regulated by TGF-β, even though TGF-β signal transduction was initiated as demonstrated by nuclear localization of Smad2/3. Analysis of transcription factors and miRNAs possibly involved in E-cadherin regulation revealed high levels of miRNAs of the miR200-family, which may contribute to the stability of E-cadherin expression in human distal tubular epithelial cells. By contrast, proximal tubular epithelial cells altered their phenotype when treated with TGF-β. They became elongated and formed three-dimensional structures. Rho-kinases were identified as modulators of TGF-β-induced morphological alterations. Non-specific inhibition of Rho-kinases resulted in stabilization of the epithelial phenotype, while partial effects were observed upon downregulation of Rho-kinase isoforms ROCK1 and ROCK2. The distinct reactivity of proximal and distal cells was retained when the cells were cultured as polarized cells. Interference with Rho-kinase signaling provides a target to counteract TGF-β-mediated mesenchymal alterations of epithelial cells, particularly in proximal tubular epithelial cells. Furthermore, primary distal tubular cells differed from cell lines by their high phenotypic stability which included constant expression of E-cadherin. Our cell culture system of primary epithelial cells is thus suitable to understand and modulate cellular remodeling processes of distinct tubular cells relevant for human renal disease.


The impact of trauma exposure and moral injury on UK military veterans: a qualitative study.
Background: Exposure to a potentially morally injurious event (PMIE) has been found to be associated with a range of adverse mental health outcomes. However, how the psychological consequences following PMIEs compare to those encountered after a traumatic, but not a PMIE, remain poorly understood. Objective: The aim was to qualitatively explore UK military veterans' responses to experiences of trauma and moral injury and the impact of such events on psychological wellbeing. Method: Thirty male veterans who reported exposure to traumatic and/or morally injurious events were recruited. Semi-structured qualitative interviews were conducted, and data were analysed using thematic analysis. Results: Six veterans described exposure to a non-morally injurious traumatic event, 15 reported experiencing a PMIE, and 9 described exposure to a 'mixed' event which was simultaneously morally injurious and traumatic. Veterans who encountered a PMIE described experiencing moral dissonance, or a clash between concurrently held sets of values (e.g. military values versus civilian values), which provoked considerable psychological distress. Veterans' cognitions and responses were found to differ following a PMIE compared to a traumatic, but not PMIE, which could have negative implications for daily functioning. Several risk and protective factors for experiencing distress following a PMIE were described. Conclusions: This study provides some of the first evidence that events experienced by UK veterans can simultaneously be morally injurious and traumatic or life-threatening as well as highlighting the process by which moral injury may occur in UK veterans. These findings illustrate the need to examine effective pathways for prevention and intervention for veterans who have experienced a morally injurious event.


Outcome of Patients Undergoing Transcatheter Implantation of Aortic Valve With Previous Mitral Valve Prosthesis (OPTIMAL) Study.
Transcatheter aortic valve replacement (TAVR) is the gold standard for severe valvular aortic stenosis in patients at high/prohibitive surgical risk. This procedure has also been used in patients with previous mitral valve (MV) prostheses, with contrasting outcomes reported. The aim of this study is to describe procedural and early outcomes of patients with previous MV prostheses undergoing TAVR. This is a retrospective registry of 154 patients with previous MV prostheses who underwent TAVR across high-volume medical centres at a mean of 11.7 ± 8.4 years after mitral surgery. Mean mitroaortic distance at computed tomography was 9.7 ± 4.8 mm. Procedural success was achieved in 150 (97.4%) patients, with reduction of aortic gradients (42.6 ± 14.2 to 10.0 ± 7.0 mm Hg; P < 0.001). Device success was achieved in 133 (86.3%) patients. MV prosthesis interference by the TAVR device was observed in 2 patients; in both, the mitroaortic distance was <5 mm, with 1 complicated by TAVR prosthesis embolization. Periprocedural complications included 4 (2.6%) cerebrovascular accidents, 10 (6.6%) major vascular complications, 22 (14.4%) severe bleedings, 1 (0.7%) myocardial infarction, and 5 (3.2%) in-hospital deaths (all cases cardiovascular or procedure related). At a median follow-up of 13.5 (interquartile range 1.0 to 36.0) months, 26 (16.9%) deaths occurred; 15 (9.7%) were cardiac related. Late fatal mitral prosthesis thromboses occurred in 2 patients. We recorded a case of fatal hemorrhagic stroke; hospital readmission was observed in 25 (16.2%) patients due to worsening heart failure. TAVR in patients with previous mitral prostheses appears to be safe and feasible, with good hemodynamic results at 30-day and at longer-term follow-up.


Polymorphism in Nucleotide Sequence of Mitochondrial Intergenic Region in Scleractinian Coral (Galaxea fascicularis).
A region of 826 bp that is unlikely to code for a protein, ribosomal RNA, or transfer RNA was identified between the cytochrome b and NADH-ubiquinone oxidoreductase chain 2 loci in the mitochondrial DNA of the scleractinian reef coral Galaxea fascicularis. Nucleotide sequences were determined in a part (625 bp) of this intergenic region in 95 individuals collected at 9 sites in the Ryukyu Archipelago in southwestern Japan. A total of 8 haplotypes were found, and a deletion of 290 bp was found in 3 of them. Significant differences were found in frequencies of the haplotypes at 3 sampling sites. The presence or absence of the deletion was highly correlated with the hard or soft morphotype. The deletion was found in the majority of hard-type colonies, but in a small fraction of soft-type individuals.


Hereditary hemorrhagic telangiectasia: diagnosis and management.
Hereditary hemorrhagic telangiectasia is an uncommon autosomal dominant disease that occurs in approximately one in 5,000 to 8,000 persons. This multisystem disorder can affect the nose, skin, gastrointestinal tract, lungs, liver, and brain. Epistaxis is the most common presenting problem, occurring in 90 percent of affected patients. Approximately 15 to 30 percent of patients with hereditary hemorrhagic telangiectasia will have an arteriovenous malformation in the lungs and more than 10 percent will have one in the brain. The symptoms of hereditary hemorrhagic telangiectasia are often unrecognized. Many patients, even those with affected family members, may go undiagnosed. Hereditary hemorrhagic telangiectasia is a clinical diagnosis that is based on the presence of three of four criteria (i.e., epistaxis, telangiectasias, visceral arteriovenous malformations, or family history of the disease). Screening and treatment recommendations have been created in an attempt to limit the morbidity and mortality associated with this disease. Patients with confirmed or suspected hereditary hemorrhagic telangiectasia should be screened for brain and lung arteriovenous malformations using magnetic resonance imaging of the brain and contrast echocardiography. Pulmonary arteriovenous malformations can be treated with embolization. Patients with a history of pulmonary arteriovenous malformations or those who have not been screened should use antibiotic prophylaxis before dental treatment, endoscopy, or other procedures that could cause bacteremia because of the risk of paradoxical brain embolism or infection.


Improved Complex Skill Acquisition by Immersive Virtual Reality Training: A Randomized Controlled Trial.
There has been limited literature on immersive virtual reality (VR) simulation in orthopaedic education. The purpose of this multicenter, blinded, randomized controlled trial was to determine the validity and efficacy of immersive VR training in orthopaedic resident education. Nineteen senior orthopaedic residents (resident group) and 7 consultant shoulder arthroplasty surgeons (expert group) participated in the trial comparing immersive VR with traditional learning using a technical journal article as a control. The examined task focused on achieving optimal glenoid exposure. Participants completed demographic questionnaires, knowledge tests, and a glenoid exposure on fresh-frozen cadavers while being examined by blinded shoulder arthroplasty surgeons. Training superiority was determined by the outcome measures of the Objective Structured Assessment of Technical Skills (OSATS) score, a developed laboratory metric, verbal answers, and time to task completion. Immersive VR had greater realism and was superior in teaching glenoid exposure than the control (p = 0.01). The expert group outperformed the resident group on knowledge testing (p = 0.04). The immersive VR group completed the learning activity and knowledge tests significantly faster (p < 0.001) at a mean time (and standard deviation) of 11 ± 3 minutes than the control group at 20 ± 4 minutes, performing 3 to 5 VR repeats for a reduction in learning time of 570%. The immersive VR group completed the glenoid exposure significantly faster (p = 0.04) at a mean time of 14 ± 7 minutes than the control group at 21 ± 6 minutes, with superior OSATS instrument handling scores (p = 0.03). The immersive VR group scored equivalently in surprise verbal scores (p = 0.85) and written knowledge scores (p = 1.0). Immersive VR demonstrated substantially improved translational technical and nontechnical skills acquisition over traditional learning in senior orthopaedic residents. Additionally, the results demonstrate the face, content, construct, and transfer validity for immersive VR. This adequately powered, randomized controlled trial demonstrated how an immersive VR system can efficiently (570%) teach a complex surgical procedure and also demonstrate improved translational skill and knowledge acquisition when compared with a traditional learning method.


Amyloid-β and hyperphosphorylated tau synergy drives metabolic decline in preclinical Alzheimer's disease.
This study was designed to test the interaction between amyloid-β and tau proteins as a determinant of metabolic decline in preclinical Alzheimer's disease (AD). We assessed 120 cognitively normal individuals with [18F]florbetapir positron emission tomography (PET) and cerebrospinal fluid (CSF) measurements at baseline, as well as [18F]fluorodeoxyglucose ([18F]FDG) PET at baseline and at 24 months. A voxel-based interaction model was built to test the associations between continuous measurements of CSF biomarkers, [18F]florbetapir and [18F]FDG standardized uptake value ratios (SUVR). We found that the synergistic interaction between [18F]florbetapir SUVR and CSF phosphorylated tau (p-tau) measurements, rather than the sum of their independent effects, was associated with a 24-month metabolic decline in basal and mesial temporal, orbitofrontal, and anterior and posterior cingulate cortices (P<0.001). In contrast, interactions using CSF amyloid-β1-42 and total tau biomarkers did not associate with metabolic decline over a time frame of 24 months. The interaction found in this study further support the framework that amyloid-β and hyperphosphorylated tau aggregates synergistically interact to cause downstream AD neurodegeneration. In fact, the regions displaying the metabolic decline reported here were confined to brain networks affected early by amyloid-β plaques and neurofibrillary tangles. Preventive clinical trials may benefit from using a combination of amyloid-β PET and p-tau biomarkers to enrich study populations of cognitively normal subjects with a high probability of disease progression in studies, using [18F]FDG as a biomarker of efficacy.


Evaluation of Three Sources of Validity Evidence for a Synthetic Thoracoscopic Esophageal Atresia/Tracheoesophageal Fistula Repair Simulator.
Thoracoscopic esophageal atresia (EA)/tracheoesophageal fistula (TEF) repair is technically challenging. We have previously reported our experiences with a high-fidelity hybrid model for simulation-based educational instruction in thoracoscopic EA/TEF, including the high cost of the tissue for these models. The purposes of this study were (1) to create a low-cost synthetic tissue EA/TEF repair simulation model and (2) to evaluate the content validity of the synthetic tissue simulator. Review of the literature and computed tomography images were used to create computer-aided drawings (CAD) for a synthetic, size-appropriate EA/TEF tissue insert. The inverse of the CAD image was then printed in six different sections to create a mold that could be filled with platinum-cured silicone. The silicone EA/TEF insert was then placed in a previously described neonatal thorax and covered with synthetic skin. Following institutional review board-exempt determination, 47 participants performed some or all of a simulated thoracoscopic EA/TEF during two separate international meetings (International Pediatric Endosurgery Group [IPEG] and World Federation of Associations of Pediatric Surgeons [WOFAPS]). Participants were identified as "experts," having 6-50 self-reported thoracoscopic EA/TEF repairs, and "novice," having 0-5 self-reported thoracoscopic EA/TEF repairs. Participants completed a self-report, six-domain, 24-item instrument consisting of 23 5-point rating scales and one 4-point Global Rating Scale. Validity evidence relevant to test content and response processes was evaluated using the many-facet Rasch model, and evidence of internal structure (interitem consistency) was estimated using Cronbach's alpha. A review of the participants' ratings indicates there were no overall differences across sites (IPEG versus WOFAPS, P=.84) or experience (expert versus novice, P=.17). The highest observed averages were 4.4 (Value of Simulator as a Training Tool), 4.3 (Physical Attributes-chest circumference, chest depth, and intercostal space), and 4.3 (Realism of Experience-fistula location). The lowest observed averages were 3.5 (Ability to Perform-closure of fistula), 3.7 (Ability to Perform-acquisition target trocar sites), 3.8 (Physical Attributes-landmark visualization), 3.8 (Ability to Perform-anastomosis and dissection of upper pouch), and 3.9 (Realism of Materials-skin). The Global Rating Scale was 2.9, coinciding with a response of "this simulator can be considered for use in neonatal TEF repair training, but could be improved slightly." Material costs for the synthetic EA/TEF inserts were less than $2 U.S. per insert. We have successfully created a low-cost synthetic EA/TEF tissue insert for use in a neonatal thoracoscopic EA/TEF repair simulator. Analysis of the participants' ratings of the synthetic EA/TEF simulation model indicates that it has value and can be used to train pediatric surgeons, especially those early in their learning curve, to begin to perform a thoracoscopic EA/TEF repair. Areas for model improvement were identified, and these areas will be the focus for future modifications to the synthetic EA/TEF repair simulator.


Sports participation and physical education in American secondary schools: current levels and racial/ethnic and socioeconomic disparities.
The purpose of this study was to determine the current levels of physical education (PE) and sports participation among American secondary school students, and to establish the extent to which they vary by grade level, racial/ethnic background, and socioeconomic status (SES) of the students. Nationally representative data were used from over 500 schools and 54,000 students surveyed in 2003, 2004, and 2005 as part of the Youth, Education, and Society (YES) study and the Monitoring the Future (MTF) study. As part of YES, school administrators completed questionnaires on physical activity (including rates of sports and PE participation) of students in their schools. Students in the same schools completed self-administered questionnaires in the same year as part of MTF, providing individual background data, including their gender, racial/ethnic identification, and parents' education level. Data were analyzed in 2006. Physical education requirements, and actual student participation rates, decline substantially between 8th and 12th grades. About 87% of 8th graders were in schools that required them to take PE, compared to only 20% of 12th graders. Principals estimate that over 90% of 8th graders actually take PE, compared to 34% of 12th graders. Subgroup differences in PE participation rates were small. Only a fraction of all students participate in varsity sports during the school year, with girls participating only slightly less than boys (33% vs 37%). Participation correlates negatively with SES and was lower among black and Hispanic students than white students, even after controlling for other variables. Participation rates in intramural sports were even lower, declined in higher grades, and were lower among low-SES and Hispanic students (after controlling for other variables). Physical education is noticeably lacking in American high schools for all groups. Racial/ethnic minorities and low-SES youth, who are at higher than average risk of being overweight in adolescence, are getting less exercise due to their lower participation in school sports. Disparities in resources available to minorities and lower-SES youth may help explain the differences in participation rates.


Inflammatory bowel disease: a survey of the epidemiology in Asia.
Inflammatory bowel disease (IBD) has long been considered a disease that affects predominantly a Western population. The incidence and prevalence rates from Asian populations are much lower in comparison. More recent data, however, have shown significantly higher rates in Asians and time trend studies have shown an increase in the incidence of ulcerative colitis (UC) and a similar but lower rise in Crohn's disease (CD). The epidemiological changes that are taking place mirror that of the Western experience seen 50 years previously and seem to occur in parallel with the rapid socioeconomic development taking place in Asia. It appears that certain racial groups are more prone than others to develop IBD. For instance, Indians in South-East Asia have higher rates compared to Chinese and Malays. While there is host genetic predisposition, environmental factor(s) may be responsible for this difference. Migrant studies of South Asians in the UK, where second-generation immigrants have assumed incidence rates as high as the indigenous whites and Asian Jews who develop high incidence rates comparable to Jews from Europe or North America in Israel point to the role of environmental factors. It is unclear which specific factors are responsible. Studies have suggested a change in diet to a more Westernized one may underlie this epidemiological change in the Asian population. It is likely that there are racial groups amongst Asians who are more susceptible to IBD and who will demonstrate a higher frequency of IBD when exposed to putative environmental factors.


Perspectives on an oral motor activity: the use of rubber tubing as a "chewy".
Among school-aged children with learning and behavioral problems, a therapeutic tool found to be helpful is a "chewy." A chewy is a cylindrical piece of rubber tubing, the ends of which are placed in the mouth and are chewed or sucked on. This behavior appears to have a calming, organizing, and focusing effect on the child. In addition, for some children who put nonedible items in their mouths, a chewy can provide an appropriate sensory substitute or alternative. A literature review supporting the benefits of oral motor input is discussed. Three case studies of children using a chewy are presented. A discussion follows regarding the therapeutic benefits of the use of a chewy and the consideration of it as part of a child's sensory diet. Important guidelines regarding the appearance, taste, length, durability, and sanitation of the chewy are reviewed.


Permissible and immunogenic HLA-A mismatches: cytotoxic T-cell precursor frequencies reflect graft survival data.
Analysis of the in vivo immunogenicity of single HLA mismatches, in the context of a patient's own human leukocyte antigen (HLA) phenotype, has been used to define permissible and immunogenic HLA mismatches. Kidney graft survival in the case of permissible mismatches was similar to that of completely HLA matched combinations, whereas immunogenic mismatches lead to a significantly poorer graft survival. The present study tested whether such permissible and immunogenic HLA mismatches are reflected in the in vitro cytotoxic T-lymphocyte (CTL) allorepertoire. Limiting dilution experiments were performed to analyze the number of precursor CTL directed against individual HLA class I antigens. In general, the frequency of CTLp directed against permissible HLA-A antigens (n = 70, mean frequency 27 CTLp per million peripheral blood lymphocytes [PBL]) was found to be significantly lower compared with the CTLp directed against immunogenic HLA-A antigens (n = 73, mean frequency 59 CTLp per million PBL). The difference was found both in healthy individuals and a population of renal transplant candidates. These results were confirmed by a retrospective analysis of CTLp frequencies performed between partly mismatched unrelated bone marrow donors and their potential recipients. In conclusion, on the population level the permissible and immunogenic HLA-A mismatches are indeed reflected in the CTL allorepertoire. However, due to the big overlap of the CTLp frequencies in these populations, the permissible or immunogenic nature of a mismatch for a particular patient should be determined on an individual basis.


Impact of residual normal metaphases in core binding factor acute myeloid leukemia.
Karyotype allows for stratification of outcomes in acute myeloid leukemia (AML) patients. Previous data suggested that the presence of residual normal cells improved the prognosis in patients with monosomy 7. The Southwest Oncology Group (SWOG) reported the impact of residual normal metaphases in AML patients with monosomal karyotype (MK) and found a similar relationship. We determined the influence of residual normal metaphases in patients with core binding factor (CBF) AML. The presence and total number of normal and abnormal metaphases were tallied for patients with CBF AML treated in 10 consecutive SWOG trials and used as a variable to determine the effect on complete remission, refractory disease, and overall survival (OS) rates. Among 113 CBF AML patients, median age of diagnosis was 45 years (range, 18-77 years), and median OS was 4 years (CI-2 years-not reached). Patients with inv(16) and no normal metaphases had improved OS compared with those with 1+ normal metaphases (P = .00005), whereas no difference was noted for patients with t(8;21). Multivariate analysis demonstrated that having cells with a normal karyotype had a negative impact on survival (HR, 2.11; 95% CI, 1.09-4.08; P = .026). This shorter survival was a consequence of a higher rate of refractory disease in older patients (OR, 1.03; 95% CI, 0.9998-1.06; P = .05) and in those with normal metaphases (HR, 1.26 95% CI, 1.04-1.51; P = .02). In patients with CBF AML, the presence of cells with normal metaphases and increasing age negatively affect the prognosis, especially in patients with inv(16).


GATA transcription factors integrate Wnt signalling during heart development.
Cardiogenesis is inhibited by canonical Wnt/beta-catenin signalling and stimulated by non-canonical Wnt11/JNK signalling, but how these two signalling pathways crosstalk is currently unknown. Here, we show that Wnt/beta-catenin signalling restricts cardiogenesis via inhibition of GATA gene expression, as experimentally reinstating GATA function overrides beta-catenin-mediated inhibition and restores cardiogenesis. Furthermore, we show that GATA transcription factors in turn directly regulate Wnt11 gene expression, and that Wnt11 is required to a significant degree for mediating the cardiogenesis-promoting function of GATA transcription factors. These results demonstrate that GATA factors occupy a central position between canonical and non-canonical Wnt signalling in regulating heart muscle formation.


An update on Legionella.
Legionella pneumophila is increasingly recognized as a significant cause of sporadic and epidemic community-acquired and nosocomial-acquired pneumonia. This review focuses on the latest literature concerning the epidemiology, pathogenesis, clinical presentation, diagnosis, and treatment of Legionnaires' disease. A significant increase in the incidence of Legionnaires' disease in the United States has been documented over the last years. L. pneumophila has recently been found to be a leading cause of community-acquired pneumonia in hospitalized and ambulatory patients in Germany. Recent studies provide insight into the understanding of the pathogenesis of Legionnaires' disease and the relevance of the formation of biofilms. Clinical manifestations of Legionnaires' disease are not specific and current diagnostic scores are of limited use. Several recent studies offer useful information concerning Legionnaires' disease in immunosuppressed patients. A systematic review of English literature performed to assess test characteristics of Legionella urinary antigen has found that the pooled sensitivity of the test was 0.74 and specificity was 0.991. Improved clinical response has been observed for patients with Legionnaires' disease treated with highly active antimicrobial agents against Legionella. Legionnaires' disease is a significant health problem in many countries. Clinical manifestations are unreliable in diagnosing Legionnaires' disease. Therefore, diagnostic laboratory tests for Legionella, including the urinary antigen test, should be applied to all patients with pneumonia. Levofloxacin (or other fluoroquinolone) or azithromycin are the current drugs of choice for treatment of Legionnaires' disease. Effective preventive strategies are needed.


Family history tools in primary care: does one size fit all?
Family health history (FHH) has potential value in many health care settings. This review discusses the potential uses of FHH information in primary care and the need for tools to be designed accordingly. We developed a framework in which the attributes of FHH tools are mapped against these different purposes. It contains 7 attributes mapped against 5 purposes. In considering different FHH tool purposes, it is apparent that different attributes become more or less important, and that tools for different purposes require different implementation and evaluation strategies. The context in which a tool is used is also relevant to its effectiveness. For FHH tools, it is unlikely that 'one size fits all', although appreciation of different purposes, users and contexts should facilitate the development of different applications from single FHH platforms.


Interspecific RNA interference of SHOOT MERISTEMLESS-like disrupts Cuscuta pentagona plant parasitism.
Infection of crop species by parasitic plants is a major agricultural hindrance resulting in substantial crop losses worldwide. Parasitic plants establish vascular connections with the host plant via structures termed haustoria, which allow acquisition of water and nutrients, often to the detriment of the infected host. Despite the agricultural impact of parasitic plants, the molecular and developmental processes by which host/parasitic interactions are established are not well understood. Here, we examine the development and subsequent establishment of haustorial connections by the parasite dodder (Cuscuta pentagona) on tobacco (Nicotiana tabacum) plants. Formation of haustoria in dodder is accompanied by upregulation of dodder KNOTTED-like homeobox transcription factors, including SHOOT MERISTEMLESS-like (STM). We demonstrate interspecific silencing of a STM gene in dodder driven by a vascular-specific promoter in transgenic host plants and find that this silencing disrupts dodder growth. The reduced efficacy of dodder infection on STM RNA interference transgenics results from defects in haustorial connection, development, and establishment. Identification of transgene-specific small RNAs in the parasite, coupled with reduced parasite fecundity and increased growth of the infected host, demonstrates the efficacy of interspecific small RNA-mediated silencing of parasite genes. This technology has the potential to be an effective method of biological control of plant parasite infection.