Malaria in the Republic of Djibouti, 1998-2009.
Historically, native populations in the Republic of Djibouti have experienced only low and unstable malaria transmission and intermittent epidemics. In recent years, efforts at malaria control have been aggressively pursued. This study was performed to inform revised malaria prevention recommendations for military service members and international travelers to the country. Laboratory-confirmed cases of malaria documented at large medical facilities and within military and civilian health care systems in the Republic of Djibouti from 1998 to 2009 were reviewed. In recent years, fewer than 5% of febrile cases among the three largest passive surveillance systems were laboratory-confirmed as malaria, and incidence of confirmed malaria was well below 1/1,000 persons/year. As efforts in the Republic of Djibouti progress toward elimination, and in conjunction with continued efforts at surveillance, emphasizing mosquito-avoidance measures and standby emergency treatment will become reasonable recommendations for malaria prevention.


High-throughput proteomic analysis of FFPE tissue samples facilitates tumor stratification.
Formalin-fixed, paraffin-embedded (FFPE), biobanked tissue samples offer an invaluable resource for clinical and biomarker research. Here, we developed a pressure cycling technology (PCT)-SWATH mass spectrometry workflow to analyze FFPE tissue proteomes and applied it to the stratification of prostate cancer (PCa) and diffuse large B-cell lymphoma (DLBCL) samples. We show that the proteome patterns of FFPE PCa tissue samples and their analogous fresh-frozen (FF) counterparts have a high degree of similarity and we confirmed multiple proteins consistently regulated in PCa tissues in an independent sample cohort. We further demonstrate temporal stability of proteome patterns from FFPE samples that were stored between 1 and 15 years in a biobank and show a high degree of the proteome pattern similarity between two types of histological regions in small FFPE samples, that is, punched tissue biopsies and thin tissue sections of micrometer thickness, despite the existence of a certain degree of biological variations. Applying the method to two independent DLBCL cohorts, we identified myeloperoxidase, a peroxidase enzyme, as a novel prognostic marker. In summary, this study presents a robust proteomic method to analyze bulk and biopsy FFPE tissues and reports the first systematic comparison of proteome maps generated from FFPE and FF samples. Our data demonstrate the practicality and superiority of FFPE over FF samples for proteome in biomarker discovery. Promising biomarker candidates for PCa and DLBCL have been discovered.


Accelerating Maternal and Child Health Gains in Papua New Guinea: Modelled Predictions from Closing the Equity Gap Using LiST.
Many priority countries in the countdown to the millennium development goals deadline are lagging in progress towards maternal and child health (MCH) targets. Papua New Guinea (PNG) is one such country beset by challenges of geographical inaccessibility, inequity and health system weakness. Several countries, however, have made progress through focused initiatives which align with the burden of disease and overcome specific inequities. This study identifies the potential impact on maternal and child mortality through increased coverage of prioritised interventions within the PNG health system. The burden of disease and health system environment of PNG was documented to inform prioritised MCH interventions at community, outreach, and clinical levels. Potential reductions in maternal and child mortality through increased intervention coverage to close the geographical equity gap were estimated with the lives saved tool. A set community-level interventions, with highest feasibility, would yield significant reductions in newborn and child mortality. Adding the outreach group delivers gains for maternal mortality, particularly through family planning. The clinical services group of interventions demands greater investment but are essential to reach MCH targets. Cumulatively, the increased coverage is estimated to reduce the rates of under-five mortality by 19 %, neonatal mortality by 26 %, maternal mortality ratio by 10 % and maternal mortality by 33 %. Modest investments in health systems focused on disadvantaged populations can accelerate progress in maternal and child survival even in fragile health systems like PNG. The critical approach may be to target interventions and implementation appropriately to the sensitive context of lagging countries.


Management and outcome of focal low-grade brainstem tumors in pediatric patients: the St. Jude experience.
Whereas diffuse intrinsic pontine gliomas generally have a short symptom duration and more cranial nerve involvement, focal brainstem gliomas are commonly low grade, with fewer cranial neuropathies. Although these phenotypic distinctions are not absolute predictors of outcome, they do demonstrate correlation in most cases. Because there is a limited literature on focal brainstem gliomas in pediatric patients, the objective of this paper was to report the management and outcome of these tumors. The authors reviewed the records of all children diagnosed with radiographically confirmed low-grade focal brainstem gliomas from 1986 to 2010. Each patient underwent biopsy or resection for tissue diagnosis. Event-free survival (EFS) and overall survival were evaluated. Univariate analysis was conducted to identify demographic and treatment variables that may affect EFS. Fifty-two patients (20 girls, 32 boys) with follow-up data were identified. Median follow-up was 10.0 years, and the median age at diagnosis was 6.5 years (range 1-17 years). The tumor locations were midbrain (n = 22, 42%), pons (n = 15, 29%), and medulla (n = 15, 29%). Surgical extirpation was the primary treatment in 25 patients (48%). The 5- and 10-year EFS and overall survival were 59%/98% and 52%/90%, respectively. An event or treatment failure occurred in 24 patients (46%), including 5 deaths. Median time to treatment failure was 3.4 years. Disease progression in the other 19 patients transpired within 25.1 months of diagnosis. Thirteen of these patients received radiation, including 11 within 2 months of primary treatment failure. Although children with intrinsic tumors had slightly better EFS at 5 years compared with those with exophytic tumors (p = 0.054), this difference was not significant at 10 years (p = 0.147). No other variables were predictive of EFS. Surgery suffices in many children with low-grade focal brainstem gliomas. Radiation treatment is often reserved for disease progression but offers comparable disease control following biopsy. In the authors' experience, combining an assessment of clinical course, imaging, and tumor biopsy yields a reasonable model for managing children with focal brainstem tumors.


Merkel Cell Carcinoma of the Buccal Mucosa and Lower Lip.
Merkel cell carcinoma (MCC) is an uncommon relatively aggressive neuroendocrine dermal neoplasm first described in 1972 as a tumor of the sun exposed skin. Although most MCC affect the skin of the head and neck, rare primarily oral mucosal cases have been documented. Merkel cells are nondendritic neuroendocrine cells that are found not only in the skin but also the oral mucosa and give rise to MCC. Neuroendocrine cells may be found as aggregates in organs or as diffuse or isolated cells within organs and their epithelial lining. They contain peptide hormones and biogenic amines and occur in two forms: dendritic, which are not associated with nerve fibers and non-dendritic, which are associated with nerve fibers. Merkel cells as well as MCC express simple epithelium-type Cytokeratins (8, 18, 19, 20), neurosecretory substances; chromogranin A, synaptophysin, neuron-specific enolase (NSE), adhesion molecules, and villin (intermediate filament). Though weakly, they also express neural markers such as S-100 protein. Cytokeratin 20, and Cluster of differentiation 56, are the two key diagnostic markers for Merkel cells and MCC. Etiology includes UV radiation, the recently described Merkel cell polyomavirus, and long term systemic immunosuppression. The cutaneous and mucosal variants of MCC are considered aggressive tumors with a high risk for local recurrence and metastasis and should be considered in the differential diagnosis of head and neck mucosal lesions. We present two cases of primary Merkel cell carcinoma, one on the buccal mucosa and the other on the lower lip, and discuss the salient histologic, immunohistochemical and clinical features.


Measurement of intrinsic properties of amyloid fibrils by the peak force QNM method.
We report the investigation of the mechanical properties of different types of amyloid fibrils by the peak force quantitative nanomechanical (PF-QNM) technique. We demonstrate that this technique correctly measures the Young's modulus independent of the polymorphic state and the cross-sectional structural details of the fibrils, and we show that values for amyloid fibrils assembled from heptapeptides, α-synuclein, Aβ(1-42), insulin, β-lactoglobulin, lysozyme, ovalbumin, Tau protein and bovine serum albumin all fall in the range of 2-4 GPa.


Effects of a standardised extract of Trifolium pratense (Promensil) at a dosage of 80mg in the treatment of menopausal hot flushes: A systematic review and meta-analysis.
To critically assess the evidence for a specific standardised extract of Trifolium pratense isoflavones (Promensil) at a dosage of 80mg/day in the treatment of menopausal hot flushes. Systematic literature searches were performed in Medline, Scopus, CINAHL Plus, Cochrane, AMED and InforRMIT and citations obtained from 1996 to March 2016. Reference lists were checked; corresponding authors contacted and the grey literature searched for additional publications. Studies were selected according to predefined inclusion and exclusion criteria. All randomised clinical trials of a specific standardised extract of Trifolium pratense isoflavones (Promensil) used as a mono-component at 80mg/day and measuring vasomotor symptoms were included. The data extraction and quality assessment were performed independently by one reviewer and validated by a second with any disagreements being settled by discussion. Weighted mean differences and 95% confidence intervals were calculated for continuous data using the fixed-effects model. Twenty potentially relevant papers were identified, with only five studies meeting the inclusion criteria. The meta-analysis demonstrated a statistical and clinically relevant reduction in hot flush frequency in the active treatment group compared to placebo. Weighted mean difference 3.63 hot flushes per day: [95% CI 2.70-4.56]; p˂0.00001). Due to a lack of homogeneity a priori defined sub-group analyses were performed demonstrating a substantive difference between cross-over and parallel-arm clinical trial designs. There is evidence for a statistical and clinically significant benefit for using a specific standardised extract of red clover isoflavones (Promensil) at 80mg/day for treating hot flushes in menopausal women across the 3 studies included in the meta-analysis. The preparation was safe over the short-term duration of the studies (3 months).


VEGF expression correlates with neuronal differentiation and predicts a favorable prognosis in patients with neuroblastoma.
Neuroblastoma (NB) is a childhood cancer with a low survival rate and great metastatic potential. Vascular endothelial growth factor (VEGF), an angiogenesis factor, has been found to be involved in CRT-related neuronal differentiation of NB cells. In this study, we further confirmed the role VEGF in NB through mouse xenograft model and clinical analysis from NB patients. In xenograft experiments, CRT overexpression effectively inhibited the tumor growth. In addition, the mRNA and protein levels of VEGF and differentiation marker GAP-43 were upregulated by induced CRT expression. However, no significant correlation between the expression level of VEGF and microvessel density was observed in human NB tumors, suggesting a novel mechanism of VEGF participating in NB tumorigenesis through an angiogenesis-independent pathway. In NB patients' samples, mRNA expression levels of CRT and VEGF were positively correlated. Furthermore, positive VEGF expression by immunostaining of NB tumors was found to correlate well with histological grade of differentiation and predicted a favorable prognosis. In conclusion, our findings suggest that VEGF is a favorable prognostic factor of NB and might affect NB tumor behavior through CRT-driven neuronal differentiation rather than angiogenesis that might shed light on a novel therapeutic strategy to improve the outcome of NB.


A bimodal pattern of InsP(3)-evoked elementary Ca(2+) signals in pancreatic acinar cells.
InsP(3)-evoked elementary Ca(2+) release events have been postulated to play a role in providing the building blocks of larger Ca(2+) signals. In pancreatic acinar cells, low concentrations of acetylcholine or the injection of low concentrations of InsP(3) elicit a train of spatially localized Ca(2+) spikes. In this study we have quantified these responses and compared the Ca(2+) signals to the elementary events shown in Xenopus oocytes. The results demonstrate, at the same concentrations of InsP(3), Ca(2+) signals consisting of one population of small transient Ca(2+) release events and a second distinct population of larger Ca(2+) spikes. The signal mass amplitudes of both types of events are within the range of amplitudes for the elementary events in Xenopus oocytes. However, the bimodal Ca(2+) distribution of Ca(2+) responses we observe is not consistent with the continuum of event sizes seen in Xenopus. We conclude that the two types of InsP(3)-dependent events in acinar cells are both elementary Ca(2+) signals, which are independent of one another. Our data indicate a complexity to the organization of the Ca(2+) release apparatus in acinar cells, which might result from the presence of multiple InsP(3) receptor isoforms, and is likely to be important in the physiology of these cells.


Current experience with applying the GRADE approach to public health interventions: an empirical study.
The Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach has been adopted by many national and international organisations as a systematic and transparent framework for evidence-based guideline development. With reference to an ongoing debate in the literature and within public health organisations, this study reviews current experience with the GRADE approach in rating the quality of evidence in the field of public health and identifies challenges encountered. We conducted semi-structured interviews with individuals/groups that have applied the GRADE approach in the context of systematic reviews or guidelines in the field of public health, as well as with representatives of groups or organisations that actively decided against its use. We initially contacted potential participants by email. Responses were obtained by telephone interview or email, and written interview summaries were validated with participants. We analysed data across individual interviews to distil common themes and challenges. Based on 25 responses, we undertook 18 interviews and obtained 15 in-depth responses relating to specific systematic reviews or guideline projects; a majority of the latter were contributed by groups within the World Health Organization. All respondents that have used the GRADE approach appreciated the systematic and transparent process of assessing the quality of the evidence. However, respondents reported a range of minor and major challenges relating to complexity of public health interventions, choice of outcomes and outcome measures, ability to discriminate between different types of observational studies, use of non-epidemiological evidence, GRADE terminology and the GRADE and guideline development process. Respondents' suggestions to make the approach more applicable to public health interventions included revisiting terminology, offering better guidance on how to apply GRADE to complex interventions and making modifications to the current grading scheme. Our findings suggest that GRADE principles are applicable to public health and well-received but also highlight common challenges. They provide a starting point for exploring options for improvements and, where applicable, testing these across different types of public health interventions. Several public health organisations are currently testing GRADE, and the GRADE Working Group is eager to engage with these groups to find ways to address concerns.


Simultaneous measurement of phase retardation and optic axis of a phase compensation film using an axially-symmetric sheared polymer network liquid crystal.
A new method for simultaneously measuring the phase retardation and optic axis of a uniaxial compensation film is demonstrated using an axially-symmetric sheared polymer network liquid crystal (SPNLC). By overlaying a tested compensation film with a calibrated SPNLC cell between crossed polarizers, two dark spots are clearly observed in a CCD image. From the orientation direction and distance of these two spots, the optic axis and phase retardation value of the compensation film can be determined. This method is particularly useful for those optical systems whose optic axis and phase retardation are dynamically changing.


GAA•TTC repeat expansion in human cells is mediated by mismatch repair complex MutLγ and depends upon the endonuclease domain in MLH3 isoform one.
DNA repeat expansion underlies dozens of progressive neurodegenerative disorders. While the mechanisms driving repeat expansion are not fully understood, increasing evidence suggests a central role for DNA mismatch repair. The mismatch repair recognition complex MutSβ (MSH2-MSH3) that binds mismatched bases and/or insertion/deletion loops has previously been implicated in GAA•TTC, CAG•CTG and CGG•CCG repeat expansion, suggesting a shared mechanism. MutSβ has been studied in a number of models, but the contribution of subsequent steps mediated by the MutL endonuclease in this pathway is less clear. Here we show that MutLγ (MLH1-MLH3) is the MutL complex responsible for GAA•TTC repeat expansion. Lentiviral expression of shRNA targeting MutL nuclease components MLH1, PMS2, and MLH3 revealed that reduced expression of MLH1 or MLH3 reduced the repeat expansion rate in a human Friedreich ataxia cell model, while targeting PMS2 did not. Using splice-switching oligonucleotides we show that MLH3 isoform 1 is active in GAA•TTC repeat expansion while the nuclease-deficient MLH3 isoform 2 is not. MLH3 isoform switching slowed repeat expansion in both model cells and FRDA patient fibroblasts. Our work indicates a specific and active role for MutLγ in the expansion process and reveals plausible targets for disease-modifying therapies.


Treatment for hepatitis C virus and cannabis use in illicit drug user patients: implications and questions.
Illicit drug users are the primary risk group for HCV transmission, and will form the largest HCV treatment population for years to come. Sylvestre et al.'s study suggests that cannabis use may benefit treatment retention and outcomes in illicit drug users undergoing HCV treatment. In fact, there is substantial evidence that cannabis use may help address key challenges faced by drug users in HCV treatment (e.g., nausea, depression), especially when such treatment occurs in the context of methadone maintenance treatment which may amplify these consequences. While further research is required on the biological and clinical aspects of the benefits of cannabis use for HCV treatment, and the effectiveness of cannabis use for HCV treatment needs to be explored in larger study populations, we advocate that in the interim existing barriers to cannabis use are removed for drug users undergoing HCV treatment until the conclusive empirical basis for evidence-based guidance is available.


European athletes dominate performances in Double Iron ultra-triathlons--a retrospective data analysis from 1985 to 2010.
We investigated the participation and performance trends of ultra-endurance triathletes from all nationalities competing in a Double Iron ultra-triathlon (7.6-km swim, 360-km cycle and 84.4-km run) from 1985 to 2010. A total of 1854 athletes participated in 92 Double Iron ultra-triathlons. The majority of the winners came from Europe with 72 victories, followed by North America with 17 victories. The race time for the European ultra-triathletes was 1340 (s=95.3) min, decreasing highly significantly (r (2)=0.28; P<0.0001) across the years. North American ultra-triathletes finished the races within 1556 (s=124.5) min; their race time showed no changes across the years (r (2)=0.045; P=0.07). The race time for the Europeans was highly significantly faster compared to the North Americans (P<0.0001). Future studies should investigate each country in Europe and North America in order to find the country with the largest participation of athletes and their best performance.


Evaluation of Actical equations and thresholds to predict physical activity intensity in young children.
Abstract This study examined the validity of current Actical activity energy expenditure (AEE) equations and intensity cut-points in preschoolers using AEE and direct observation as criterion measures. Forty 4-6-year-olds (5.3 ± 1.0 years) completed a ~150-min room calorimeter protocol involving age-appropriate sedentary behaviours (SBs), light intensity physical activities (LPAs) and moderate-to-vigorous intensity physical activities (MVPAs). AEE and/or physical activity intensity were calculated using Actical equations and cut-points by Adolph, Evenson, Pfeiffer and Puyau. Predictive validity was examined using paired sample t-tests. Classification accuracy was evaluated using weighted kappas, sensitivity, specificity and area under the receiver operating characteristic curve. The Pfeiffer equation significantly overestimated AEE during SB and underestimated AEE during LPA (P < 0.0125 for both). There was no significant difference between measured and predicted AEEs during MVPA. The Adolph cut-point showed significantly higher accuracy for classifying SB, LPA and MVPA than all others. The available Actical equation does not provide accurate estimates of AEE across all intensities in preschoolers. However, the Pfeiffer equation performed reasonably well for MVPA. Using cut-points of ≤6 counts · 15 s(-1), 7-286 counts · 15 s(-1) and ≥ 287 counts · 15 s(-1) when classifying SB, LPA and MVPA, respectively, is recommended.


Heat reactivation of the alpha-hemolytic, dermonecrotic, lethal activities of crude and purified staphylococcal alpha-toxin.
Manohar, M. (University of Minnesota, St. Paul), S. Kumar, and R. K. Lindorfer. Heat reactivation of the alpha-hemolytic, dermonecrotic, and lethal activities of crude and purified staphylococcal alpha-toxin. J. Bacteriol. 91:1681-1685. 1966.-Crude staphylococcal toxin loses its alpha-hemolytic activity more rapidly at 60 than at 100 C. This paradoxical behavior has been postulated to be due to the presence of a thermolabile inhibitor in crude toxin. This work provides experimental evidence for the presence of a thermolabile "protective inhibitor." This substance(s) protects the alpha-toxin against destruction at 60 C, yet simultaneously inhibits the hemolytic activity of alpha-toxin under the same conditions. Of greater importance, this work also demonstrates that the dermonecrotic and lethal activities of crude toxin are inactivated and reactivated in parallel with the alpha-hemolytic activity. Crude staphylococcal toxin possessing a high alpha-hemolytic titer when heated to 60 C for 30 min lost its alpha-hemolytic, dermonecrotic, and lethal activity. However, when this same toxin was immediately exposed to 100 C, a remarkable simultaneous reactivation of all three of these activities occurred. Contrariwise, electrophoretically purified alpha-hemolysin, which also possessed dermonecrotic and lethal activity, showed no reactivation under these conditions, thus demonstrating that reactivation is due to a substance(s) distinct from the alpha-toxin. The fact that alpha-hemolytic, dermonecrotic, and lethal activities were inactivated at 60 C and simultaneously reactivated at 100 C provides additional proof that these activities are all associated with one toxic component. The probability is remote that three separate entities would exhibit the same rate of inactivation and the same strange reactivation.


Akt/PKB regulates hepatic metabolism by directly inhibiting PGC-1alpha transcription coactivator.
Type 2 diabetes mellitus, a disease with significant effects on the health and economy of Western societies, involves disturbances in both lipid and carbohydrate metabolism. In the insulin-resistant or diabetic state, the liver is unresponsive to the actions of insulin with regard to the suppression of glucose output but continues to produce large amounts of lipid, the latter mimicking the fed, insulin-replete condition. The disordered distribution of lipids contributes to the cardiovascular disease that is the greatest cause of mortality of type 2 diabetes mellitus. Yet the precise signal transduction pathways by which insulin regulates hepatic lipid synthesis and degradation remain largely unknown. Here we describe a mechanism by which insulin, through the intermediary protein kinase Akt2/protein kinase B (PKB)-beta, elicits the phosphorylation and inhibition of the transcriptional coactivator peroxisome proliferator-activated receptor-coactivator 1alpha (PGC-1alpha), a global regulator of hepatic metabolism during fasting. Phosphorylation prevents the recruitment of PGC-1alpha to the cognate promoters, impairing its ability to promote gluconeogenesis and fatty acid oxidation. These results define a mechanism by which insulin controls lipid catabolism in the liver and suggest a novel site for therapy in type 2 diabetes mellitus.


Excess weight and gastrointestinal symptoms in Chilean celiac patients at the time of diagnosis.
celiac disease is an immune condition that results in histologic changes in the small bowel and produces both digestive and extra-digestive symptoms. Intestinal damage results in malabsorption and impaired weight or impaired optimal weight gain. However, these patients may be overweight or obese in spite of histologic damage. The aim of this study was to determine the prevalence of excess weight in newly diagnosed (adult) celiac patients. this was a retrospective observational study of patients recently diagnosed with celiac disease according to the standard Marsh classification. Nutritional status was assessed based on body mass index (BMI), as categorized by the World Health Organization (WHO). Clinical presentation was classified as typical or atypical. Potential differences in gastrointestinal symptoms according to nutritional status were also assessed. a total of 135 medical records of adult celiac patients (women = 123; men = 12) were reviewed. The average weight and BMI were 61.1 kg and 23.7 kg/m2, respectively. The proportion of typical clinical presentations was 59.2% and of atypical presentations 40.8%. A total of 71.8% of patients had a BMI indicating low or normal weight and 28.1% had a BMI indicative of being overweight or obese. No differences with regard to the presence of gastrointestinal symptoms were found according to nutritional status. further studies are needed to jointly assess energy intake and intestinal absorption in these patients, in order to explain the high rate of excess weight.


Co-expression of metabotropic glutamate receptor type 1alpha with homer-1a/Vesl-1S increases the cell surface expression of the receptor.
Homer-1a is a 30 kDa protein that forms part of a family of conserved Homer-related proteins that interact with the C-termini of the metabotropic glutamate receptors mGluR1alpha and mGluR5a. Analysis of HEK-293 cells by PCR showed that they contained mRNA coding for members of the Homer family with the predominant form being Homer-1b, which is consistent with the immunochemical analysis of these cells. Homer-1a could not be detected by immunochemical analysis. To examine the function of Homer-1a, HEK-293 cells were transfected with cDNA encoding mGluR1alpha or Homer-1a or co-transfected with both cDNAs. When cells were co-transfected with the cDNAs for both proteins, immunofluorescent staining and biotinylation of cell surface molecules revealed a significant increase in the amount of receptor present at the cell surface in contrast to cells transfected with mGluR1alpha cDNA alone. This finding was consistent with a concomitant increase in the production of inositol phosphates after treatment of the doubly transfected cells with agonist. Intracellular immunostaining for both proteins revealed that they were co-localized and underwent a redistribution into a large vesicular compartment when they were co-expressed.


Legionella longbeachae detected in an industrial cooling tower linked to a legionellosis outbreak, New Zealand, 2015; possible waterborne transmission?
A legionellosis outbreak at an industrial site was investigated to identify and control the source. Cases were identified from disease notifications, workplace illness records, and from clinicians. Cases were interviewed for symptoms and risk factors and tested for legionellosis. Implicated environmental sources were sampled and tested for legionella. We identified six cases with Legionnaires' disease and seven with Pontiac fever; all had been exposed to aerosols from the cooling towers on the site. Nine cases had evidence of infection with either Legionella pneumophila serogroup (sg) 1 or Legionella longbeachae sg1; these organisms were also isolated from the cooling towers. There was 100% DNA sequence homology between cooling tower and clinical isolates of L. pneumophila sg1 using sequence-based typing analysis; no clinical L. longbeachae isolates were available to compare with environmental isolates. Routine monitoring of the towers prior to the outbreak failed to detect any legionella. Data from this outbreak indicate that L. pneumophila sg1 transmission occurred from the cooling towers; in addition, L. longbeachae transmission was suggested but remains unproven. L. longbeachae detection in cooling towers has not been previously reported in association with legionellosis outbreaks. Waterborne transmission should not be discounted in investigations for the source of L. longbeachae infection.


Genetic variants associated with response to lithium treatment in bipolar disorder: a genome-wide association study.
Lithium is a first-line treatment in bipolar disorder, but individual response is variable. Previous studies have suggested that lithium response is a heritable trait. However, no genetic markers of treatment response have been reproducibly identified. Here, we report the results of a genome-wide association study of lithium response in 2563 patients collected by 22 participating sites from the International Consortium on Lithium Genetics (ConLiGen). Data from common single nucleotide polymorphisms (SNPs) were tested for association with categorical and continuous ratings of lithium response. Lithium response was measured using a well established scale (Alda scale). Genotyped SNPs were used to generate data at more than 6 million sites, using standard genomic imputation methods. Traits were regressed against genotype dosage. Results were combined across two batches by meta-analysis. A single locus of four linked SNPs on chromosome 21 met genome-wide significance criteria for association with lithium response (rs79663003, p=1·37 × 10(-8); rs78015114, p=1·31 × 10(-8); rs74795342, p=3·31 × 10(-9); and rs75222709, p=3·50 × 10(-9)). In an independent, prospective study of 73 patients treated with lithium monotherapy for a period of up to 2 years, carriers of the response-associated alleles had a significantly lower rate of relapse than carriers of the alternate alleles (p=0·03268, hazard ratio 3·8, 95% CI 1·1-13·0). The response-associated region contains two genes for long, non-coding RNAs (lncRNAs), AL157359.3 and AL157359.4. LncRNAs are increasingly appreciated as important regulators of gene expression, particularly in the CNS. Confirmed biomarkers of lithium response would constitute an important step forward in the clinical management of bipolar disorder. Further studies are needed to establish the biological context and potential clinical utility of these findings. Deutsche Forschungsgemeinschaft, National Institute of Mental Health Intramural Research Program.


Atmospheric deposition of polychlorinated dibenzo-p-dioxins and dibenzofurans on the soils in the vicinity of municipal solid waste incinerators.
This study focuses on the atmospheric deposition of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) on the soils in the vicinity of two municipal solid waste incinerators (MSWIs), which were located in the Gangshan (GS) and the Renwu (RW) Townships of southern Taiwan. PCDD/Fs in the soils were sampled simultaneously with those in the ambient air and analyzed for 17 2,3,7,8-substituted PCDD/Fs. The results show that the mean contents of PCDD/Fs in the soils near MSWI-GS and MSWI-RW were 2.65 and 1.20 ng I-TEQ/kg dry weight, respectively. Annual wet deposition fluxes of total PCDD/Fs were 119 and 113 ng/m(2)-year in the ambient air near MSWI-GS and MSWI-RW, respectively. The results obtained in this study are much higher than those estimated for the Atlantic Ocean, where the average wet deposition was only 45 ng/m(2)-year. The annual dry deposition fluxes accounted for 58.2 and 66.7%, respectively, indicating that dry deposition was more dominant than wet deposition in the atmospheric deposition processes. The congener profiles of 17 2,3,7,8-substituted PCDD/Fs showed that OCDD dominates in the soils. The contributions of OCDD in the soils near MSWI-GS and MSWI-RW were 73.4 and 67.1%, respectively, while they were only 41.4 and 31.2% in the atmospheric deposition, respectively. These results imply that OCDD is more persistent in the environment than other congeners. The results of the present study strongly suggest that exposure to PCDD/Fs in these areas should be reduced.


Assessment of indeterminate pulmonary nodules detected in lung cancer screening: Diagnostic accuracy of FDG PET/CT.
A major drawback of lung cancer screening programs is the high frequency of false-positive findings on computed tomography (CT). We investigated the accuracy of selective 2-[fluorine-18]-fluoro-2-deoxy-d-glucose (FDG) Positron Emission Tomography/Computed Tomography (PET/CT) scan in assessing radiologically indeterminate lung nodules detected in lung cancer screening. FDG PET/CT was performed to characterize 64 baseline lung nodules >10mm and 36 incidence nodules detected on low-dose CT screening in asymptomatic current or former smokers (83 men, age range 40-83 years) at high risk for lung cancer. CT images were acquired without intravenous contrast. Nodules were analyzed by size, density, and metabolic activity and visual scored on a 5-point scale for FDG uptake. Nodules were classified as negative for malignancy when no FDG uptake was observed, or positive when focal uptake was observed in the visual analysis, and the maximum standardized uptake value (SUVmax) was measured. Final diagnosis was based on histopathological evaluation or at least 24 months of follow-up. A total of 100 nodules were included. The prevalence of lung cancer was 1%. The sensitivity, specificity, NPV and PPV of visual analysis to detect malignancy were 84%, 95%, 91%, and 91%, respectively, with an accuracy of 91% (AUC 0.893). FDG PET/CT accurately detected 31 malignant tumors (diameters 9-42mm, SUVmax range 0.6-14.2) and was falsely negative in 6 patients. With SUVmax thresholds for malignancy of 1.5, 2, and 2.5, specificity was 97% but sensitivity decreased to 65%, 49%, and 46% respectively, and accuracy decreased to 85%, 79%, and 78% respectively (AUC 0.872). The visual analysis of FDG PET/CT scan is highly accurate in characterizing indeterminate pulmonary nodules detected in lung cancer screening with low-dose CT. Semi-quantitative analysis does not improve the accuracy of FDG PET/CT over that obtained with a qualitative method for lung nodule characterization.


Ad hoc category restructuring.
Participants learned to classify seemingly arbitrary words into categories that also corresponded to ad hoc categories (see, e.g., Barsalou, 1983). By adapting experimental mechanisms previously used to study knowledge restructuring in perceptual categorization, we provide a novel account of how experimental and preexperimental knowledge interact. Participants were told of the existence of the ad hoc categories either at the beginning or the end of training. When the ad hoc labels were revealed at the end of training, participants switched from categorization based on experimental learning to categorization based on preexperimental knowledge in some, but not all, circumstances. Important mediators of the extent of that switch were the amount of performance error experienced during prior learning and whether or not prior knowledge was in conflict with experimental learning. We present a computational model of the trade-off between preexperimental knowledge and experimental learning that accounts for the main results.


Marginal and internal discrepancies related to margin design of ceramic crowns fabricated by a CAD/CAM system.
The aim of this study was to evaluate the marginal discrepancy (MD) and internal discrepancy (ID) of ceramic crowns manufactured by a CAD/CAM system, having different finish lines. The hypotheses tested were that the finish line type would not influence the MD or ID of the crowns, and ID would not change in different regions. Three aluminum master dies (height: 5.5 mm, Ø: 7.5 mm, conicity: 6°) with different finish lines (TC: tilted chamfer; LC: large chamfer; RS: rounded shoulder) were manufactured. Ten impressions were made from each master die using a modified parallelometer. Impressions were poured in type IV dental stone, and 30 ceramic crowns (IPS Empress CAD, Ivoclar) were subsequently milled. The crowns were fixed on their respective metallic die using a metallic fixation device. The distance between the external edges of the crown to the edge of the cervical preparation was performed at 50 points on the respective metallic die (MD analysis). With the replica technique, the ID values of each crown were further evaluated at 12 points equidistant to each other in three regions: radius (R), axial (A), and occlusal (Occl). The measurements were performed using an optical microscope (250×). The data (μm) were analyzed using ANOVA and Tukey's test (5%). The RS group (28.24 ± 11.42 μm) showed significantly lower MD values (p= 0.001) than those of TC (99.92 ± 18.32 μm) and LC (64.71 ± 25.64 μm) groups, both of which also differed statistically from one another. The ID results demonstrated significantly lower values in the LC group (183.01 ± 62.82 μm) (p= 0.0014) than those of TC (216.26 ± 83.23 μm) and RS (219.12 ± 87.24 μm) groups. ID results of TC and RS were not significantly different. Additionally, the ID results showed significant differences among the regions (p= 0.0001). The null hypotheses were rejected. The RS finish line produced MD values significantly lower than tilted and large chamfer, but large chamfer presented the lowest internal discrepancy values. Independent of the finish line type, internal discrepancy was the lowest in the axial region followed by radius and occlusal regions.


Thyroid function and body weight: a community-based longitudinal study.
Body weight and overt thyroid dysfunction are associated. Cross-sectional population-based studies have repeatedly found that thyroid hormone levels, even within the normal reference range, might be associated with body weight. However, for longitudinal data, the association is less clear. Thus, we tested the association between serum thyrotropin (TSH) and body weight in a community-based sample of adult persons followed for 11 years. A random sample of 4,649 persons aged 18-65 years from a general population participated in the DanThyr study in 1997-8. We included 2,102 individuals who participated at 11-year follow-up, without current or former treatment for thyroid disease and with measurements of TSH and weight at both examinations. Multiple linear regression models were used, stratified by sex and adjusted for age, smoking status, and leisure time physical activity. Baseline TSH concentration was not associated with change in weight (women, P = 0.17; men, P = 0.72), and baseline body mass index (BMI) was not associated with change in TSH (women, P = 0.21; men, P = 0.85). Change in serum TSH and change in weight were significantly associated in both sexes. Weight increased by 0.3 kg (95% confidence interval [CI] 0.1, 0.4, P = 0.005) in women and 0.8 kg (95% CI 0.1, 1.4, P = 0.02) in men for every one unit TSH (mU/L) increase. TSH levels were not a determinant of future weight changes, and BMI was not a determinant for TSH changes, but an association between weight change and TSH change was present.


Genomic markers of panitumumab resistance including ERBB2/ HER2 in a phase II study of KRAS wild-type (wt) metastatic colorectal cancer (mCRC).
A prospective study was conducted to identify biomarkers associated with resistance to panitumumab monotherapy in patients with metastatic colorectal cancer (mCRC). Patients with previously treated, codon 12/13 KRAS wt, mCRC were prospectively administered panitumumab 6 mg/kg IV q2weeks. Of 34 panitumumab-treated patients, 11 (32%) had progressive disease at 8 weeks and were classified as non-responders. A Nanostring nCounter-based assay identified a 5-gene expression signature (ERBB2, MLPH, IRX3, MYRF, and KLK6) associated with panitumumab resistance (P = 0.001). Immunohistochemistry and in situ hybridization determined that the HER2 (ERBB2) protein was overexpressed in 4/11 non-responding and 0/21 responding cases (P = 0.035). Two non-responding tumors had ERBB2 gene amplification only, and one demonstrated both ERBB2 amplification and mutation. A non-codon 12/13 KRAS mutation occurred in one panitumumab-resistant patient and was mutually exclusive with ERBB2/HER2 abnormalities. This study identifies a 5-gene signature associated with non-response to single agent panitumumab, including a subgroup of non-responders with evidence of aberrant ERBB2/HER2 signaling. KRAS wt tumors resistant to EGFRi may be identified by gene signature analysis, and the HER2 pathway plays an important role in resistance to therapy.


Trolley Dilemmas Fail to Predict Ethical Judgment in a Hypothetical Vaccination Context.
We investigated whether the responses of 68 ethics committee members and staff to trolley dilemmas could predict their responses to research ethics problems concerning vaccine trials. Trolley dilemmas deal with the issue of sacrificing some for the benefit of many, which is also a core issue in the vaccination trial dilemmas. The subjects' responses to trolley dilemmas showed no statistically significant correlation with their responses to our vaccination trial dilemmas. We concluded that, if there is a component of transferable intuition between the contexts, it must be small and dominated by other factors. Furthermore, the willingness to sacrifice some for many was larger in the trolley context, despite a more favorable risk/reward ratio and the voluntary participation of the subjects at risk in the vaccination situations. We conclude that one's general willingness to trade lives in the trolley context may be an artifact that is due to its unrealistic setting.


Ethical challenges when using coercion in mental healthcare: A systematic literature review.
To better understand the kinds of ethical challenges that emerge when using coercion in mental healthcare, and the importance of these ethical challenges, this article presents a systematic review of scientific literature. A systematic search in the databases MEDLINE, PsychInfo, Cinahl, Sociological Abstracts and Web of Knowledge was carried out. The search terms derived from the population, intervention, comparison/setting and outcome. A total of 22 studies were included. Ethical considerations: The review is conducted according to the Vancouver Protocol. There are few studies that study ethical challenges when using coercion in an explicit way. However, promoting the patient's best interest is the most important justification for coercion. Patient autonomy is a fundamental challenge facing any use of coercion, and some kind of autonomy infringement is a key aspect of the concept of coercion. The concepts of coercion and autonomy and the relations between them are very complex. When coercion is used, a primary ethical challenge is to assess the balance between promoting good (beneficence) and inflicting harm (maleficence). In the included studies, findings explicitly related to justice are few. Some studies focus on moral distress experienced by the healthcare professionals using coercion. There is a lack of literature explicitly addressing ethical challenges related to the use of coercion in mental healthcare. It is essential for healthcare personnel to develop a strong awareness of which ethical challenges they face in connection with the use of coercion, as well as challenges related to justice. How to address ethical challenges in ways that prevent illegitimate paternalism and strengthen beneficent treatment and care and trust in connection with the use of coercion is a 'clinical must'. By developing a more refined and rich language describing ethical challenges, clinicians may be better equipped to prevent coercion and the accompanying moral distress.


Bone marrow-derived AXL tyrosine kinase promotes mitogenic crosstalk and cardiac allograft vasculopathy.
Cardiac Allograft Vasculopathy (CAV) is a leading contributor to late transplant rejection. Although implicated, the mechanisms by which bone marrow-derived cells promote CAV remain unclear. Emerging evidence implicates the cell surface receptor tyrosine kinase AXL to be elevated in rejecting human allografts. AXL protein is found on multiple cell types, including bone marrow-derived myeloid cells. The causal role of AXL from this compartment and during transplant is largely unknown. This is important because AXL is a key regulator of myeloid inflammation. Utilizing experimental chimeras deficient in the bone marrow-derived Axl gene, we report that Axl antagonizes cardiac allograft survival and promotes CAV. Flow cytometric and histologic analyses of Axl-deficient transplant recipients revealed reductions in both allograft immune cell accumulation and vascular intimal thickness. Co-culture experiments designed to identify cell-intrinsic functions of Axl uncovered complementary cell-proliferative pathways by which Axl promotes CAV-associated inflammation. Specifically, Axl-deficient myeloid cells were less efficient at increasing the replication of both antigen-specific T cells and vascular smooth muscle cells (VSMCs), the latter a key hallmark of CAV. For the latter, we discovered that Axl-was required to amass the VSMC mitogen Platelet-Derived Growth Factor. Taken together, our studies reveal a new role for myeloid Axl in the progression of CAV and mitogenic crosstalk. Inhibition of AXL-protein, in combination with current standards of care, is a candidate strategy to prolong cardiac allograft survival.


An optical power combiner/wavelength demultiplexing module for hybrid WDM FTTX.
An optical device scheme that serves simultaneously as a power combiner for upstream and wavelength demultiplexer for downstream signals is presented. The design concept is validated experimentally by an optical module based on off-the-shelf discrete optical components. An integrated device based on planar lightwave circuit (PLC) is proposed and analyzed in which a multi-mode interference (MMI) device is utilized to separate the upstream 1310 nm signal from the downstream 155x nm signals. The dense WDM function is realized through an arrayed-waveguide-grating (AWG). Design guidelines and optimization procedure for the device are discussed by way of examples.


Effects of switching from olanzapine, quetiapine, and risperidone to aripiprazole on 10-year coronary heart disease risk and metabolic syndrome status: results from a randomized controlled trial.
This study examined the clinical significance of switching from olanzapine, quetiapine, or risperidone to aripiprazole by examining changes in predicted risk of cardiovascular disease (CVD) according to the Framingham Risk Score (FRS) and metabolic syndrome status. FRS estimates 10-year risk of "hard" coronary heart disease (CHD) outcomes (myocardial infarction and coronary death) while metabolic syndrome is associated with increased risk of CVD, stroke, and diabetes mellitus. Changes in FRS and metabolic syndrome status were compared between patients with BMI ≥ 27 and non-HDL-C ≥ 130 mg/dL randomly assigned to stay on stable current treatment (olanzapine, quetiapine, or risperidone) or switch to treatment with aripiprazole with 24 weeks of follow-up. All study participants were enrolled in a behavioral program that promoted healthy diet and exercise. The pre-specified analyses included 89 switchers and 98 stayers who had post-baseline measurements needed to assess changes. Least squares mean estimates of 10-year CHD risk decreased more for the switch (from 7.0% to 5.2%) than the stay group (from 7.4% to 6.4%) (p = 0.0429). The odds ratio for having metabolic syndrome (stay vs. switch) at the last observation was 1.748 (95% CI 0.919, 3.324, p = 0.0885). Switching from olanzapine, quetiapine, or risperidone to aripiprazole was associated with larger reductions in predicted 10-year risk of CHD than the behavioral program alone. The advantage of switching on metabolic syndrome was not statistically significant. The benefits of switching must be balanced against its risks, which in this study included more discontinuations of the study treatment but no significant increase in symptoms or hospitalizations.


Efficacy and biocompatibility of a photopolymerized, synthetic, absorbable hydrogel as a dural sealant in a canine craniotomy model.
A canine craniotomy model was used to evaluate the dural sealing efficacy and biocompatibility of a novel, synthetic, bioresorbable hydrogel. Bilateral craniotomies were performed in 24 dogs assigned to six survival periods. In each animal a parasagittal durotomy was created and then repaired. At the treatment sites the hydrogel sealant was applied over the dural repair and photopolymerized. The repair was tested for leaks to 20 cm H2O by using a Valsalva maneuver. At the control sites the incisions were sutured and tested for leaks only. After uneventful survival periods, the leak test was repeated in three of the four animals in each group. Bone-dura adhesion was evaluated, after which the dura and underlying brain were removed, fixed, and examined histologically. En bloc histological investigation was performed on a specimen obtained from the fourth animal in each group. Over a 56-day period, 18 treated sites were tested for leaks. A leak was detected at a site remote from that of the repair in one animal; this was excluded from analysis. Thus 17 of 17 treated sites remained free of leaks. On the control side of one animal, there was a leak from a new dural tear at the cranial end of the durotomy, which occurred when the bone flap was removed. This site was also excluded from analysis. Eleven of 17 leak-tested control sites remained free of leaks over the study period. Bone-dura adhesions occurred in 15 of 19 control sites and had a mean adhesion score of 1.37 (range 0-4), whereas adhesions occurred in 10 of 19 treated sites with a mean adhesion score of 0.84 (range 0-3). No cortical reaction was noted. This novel hydrogel sealant is efficacious in sealing dural repair sites measuring up to 2 mm. Healing of the underlying dura is not compromised and exposed cortical tissue is not altered histologically.


Thermal Transport in Quasi-1D van der Waals Crystal Ta2Pd3Se8 Nanowires: Size and Length Dependence.
Van der Waals (vdW) crystals with covalently bonded building blocks assembled together through vdW interactions have attracted tremendous attention recently because of their interesting properties and promising applications. Compared to the explosive research on two-dimensional (2D) vdW materials, quasi-one-dimensional (quasi-1D) vdW crystals have received considerably less attention, while they also present rich physics and engineering implications. Here we report on the thermal conductivity of exfoliated quasi-1D Ta2Pd3Se8 vdW nanowires. Interestingly, even though the interatomic interactions along each molecular chain are much stronger than the interchain vdW interactions, the measured thermal conductivity still demonstrates a clear dependence on the cross-sectional size up to >110 nm. The results also reveal that partial ballistic phonon transport can persist over 13 μm at room temperature along the molecular chain direction, the longest experimentally observed ballistic transport distance with observable effects on thermal conductivity so far. First-principles calculations suggest that the ultralong ballistic phonon transport arises from the highly focused longitudinal phonons propagating along the molecular chains. These data help to understand phonon transport through quasi-1D vdW crystals, facilitating various applications of this class of materials.


Autoradiographic localization of peripheral M1 muscarinic receptors using [3H]pirenzepine.
A novel antimuscarinic agent, pirenzepine, has been proposed to distinguish at least two subtypes of muscarinic receptor. M1 receptors have been designated as those displaying a high affinity for pirenzepine. Both functional and binding studies have revealed a prevalence of M1 receptors in sympathetic ganglia while autonomic effector tissues have only low densities of M1 receptors. In the present study, in vitro autoradiographic procedures have been used to localize specifically high affinity binding sites for pirenzepine (M1 receptors) in sections of guinea-pig ileum, rat superior cervical ganglion and rat submaxillary gland. The overall localization of muscarinic receptors was also studied using the non-selective antagonist, [3H]N-methylscopolamine. The highest densities of M1 receptors were found in superior cervical ganglion, sympathetic nerve bundles, myenteric ganglia and mucous secreting cells of the submaxillary gland, while lower densities were found in smooth muscle and serous secreting cells of the submaxillary gland. No area found to possess muscarinic receptors was devoid of M1 receptors.


Phenotypic stress response of Pseudomonas aeruginosa following culture in water microcosms.
The purpose of the present study was to explore the potential behavioural changes of Pseudomonas aeruginosa following growth in different aquatic environmental conditions. To achieve this, P. aeruginosa was cultured in various water microcosms for 12 months under fixed (pH, nutrients and temperature) factors. P. aeruginosa responses to these conditions were investigated using colony morphotype, biochemical and enzymatic characterisation, pyocin typing, serotyping, sensitivity to different classes of antibiotics and molecular identification. Results show that starvation in water microcosms lead to unusual phenotypes. Of interest is that the pyocin changed from 24/n in the wild type to 83/a following culture in the water microcosms, and the serotype changed from O6 in the wild type to O1 in microcosm-cultured P. aeruginosa. Furthermore, the starvation period in various aquatic microcosms enhanced the resistance of P. aeruginosa against beta-lactam antibiotics. Compared to the other aquatic environments, the seawater microcosm produced the greatest amount of variations in P. aeruginosa. Overall, data demonstrated a high adaptability of P. aeruginosa to environmental changes. This may explain the unusual antibiotic-resistant phenotypes belonging to P. aeruginosa species, and their capacity for spreading that leads to human infections.


The effects of extracorporeal shock wave therapy on pain and range of motion in patients with adhesive capsulitis.
[Purpose] This study examined the effects of extracorporeal shock wave therapy on pain and range of motion in patients with adhesive capsulitis. [Subjects and Methods] Thirty patients with adhesive capsulitis were divided into an experimental group (n=15) that would be treated with extracorporeal shock wave therapy and a control group (n=15) that would be treated only with conservative physical therapy. Both groups were treated three times a week over a four-week period. [Results] An intra-group comparison before and after the treatment showed that both groups experienced a decline in pain and an increase in their range of motion that were statistically significant. An inter-group comparison after the treatment showed that the experimental group had a lower level of pain and a higher range of motion than the control group that were statistically significant. [Conclusion] Extracorporeal shock wave therapy may be an effective intervention for reducing pain and improving the range of motion in patients with adhesive capsulitis.


The reporting of sensitive behavior by adolescents: a methodological experiment in Kenya.
Does audio computer-assisted self-interviewing (ACASI) produce more valid reporting of sexual activity and related behaviors than face-to-face interviews or self-administered interviews? This analysis, based on data collected from over 6,000 unmarried adolescents in two districts of Kenya--Nyeri and Kisumu--indicates substantial and significant differences in reported rates of premarital sex across interview modes, although not always in the expected direction. Our assumption that girls underreport sexual activity in face-to-face interviews by comparison with ACASI is not confirmed by the Nyeri data, but our results from Kisumu are considerably more promising. As for boys, who we believe exaggerate their level of sexual activity in face-to-face interviews, a more nuanced set of expectations regarding the reporting of sensitive behaviors was offered; our results from Kisumu, although not always significant, by and large conform to expectations.


Safety of Intra-articular Hyaluronic Acid Injections in Osteoarthritis: Outcomes of a Systematic Review and Meta-Analysis.
Some controversy exists regarding the safety of intra-articular hyaluronic acid (IAHA) in the management of osteoarthritis (OA). The objective of this study was to re-assess the safety profile of IAHA in patients with OA, through a comprehensive meta-analysis of randomized, placebo-controlled trials. A comprehensive literature search was undertaken in the databases MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), and Scopus. Randomized, double-blind, placebo-controlled, parallel-group trials that assessed adverse events (AEs) with IAHA in patients with OA were eligible for inclusion. Authors and/or study sponsors were contacted to obtain the full report of AEs. The primary outcomes were overall severe and serious AEs, as well as the following MedDRA System Organ Class (SOC)-related AEs: gastrointestinal, cardiac, vascular, respiratory, nervous system, skin and subcutaneous tissue disorders, musculoskeletal, renal and urinary disorders, infections and infestations, and hypersensitivity reaction. Database searches initially identified 1481 records. After exclusions according to the selection criteria, 22 studies were included in the qualitative synthesis, and nine studies having adequate data were ultimately included in the meta-analysis. From the studies excluded according to the pre-specified selection criteria, 21 with other pharmacological OA treatments permitted during the trials were a posteriori included in a parallel qualitative synthesis, from which eight studies with adequate data were finally included in a parallel meta-analysis. Since this meta-analysis was designed to assess safety, the exclusion criterion on concomitant anti-OA medication was crucial. However, due to the high number of studies that allowed mainly concomitant oral non-steroidal anti-inflammatory drugs (NSAIDs), we decided to include them in a post hoc parallel analysis in order to compare the results from the two analyses. No statistically significant difference in odds was found between IAHA and placebo for all types of SOC-related disorders, except for infections and infestations, for which significantly lower odds were found with IAHA compared with placebo, both overall (odds ratio [OR] = 0.61, 95% confidence interval [CI] 0.40-0.93; I2 = 0%) and in studies without concomitant anti-OA medication (OR = 0.49, 95% CI 0.27-0.89). There were significant increased odds of reporting serious AEs with IAHA compared with placebo, both overall (OR = 1.78, 95% CI 1.21-2.63; I2 = 0%) and in studies with concomitant anti-OA medication (OR = 1.78, 95% CI 1.10-2.89), but not in studies without concomitant anti-OA medication (OR = 1.78, 95% CI 0.92-3.47). Using the available data on studies without any concomitant anti-OA medication permitted during clinical trials, IAHA seems not to be associated with any safety issue in the management of OA. However, this evidence was associated with only a "low" to "moderate" certainty. A possible association with increased risk of serious AEs, particularly when used with concomitant OA medications, requires further investigation.


An investigation into the perceived value of the College of Radiographers voluntary accreditation scheme for advanced and consultant practitioners in breast imaging.
A voluntary accreditation scheme has been introduced, requiring advanced (AdP) and consultant practitioners (CP) to submit several pieces of work to the College of Radiographers (CoR). However, few individuals have opted to become voluntary accredited. This study investigated the reasons behind becoming voluntary accredited, the value that was gained and why there appears to be a lack of support for the scheme. An online electronic survey was conducted using a mixed methods approach. Open questions enabled individual opinions and thoughts to be expressed, Likert scale style questions allowed further understanding of the level of agreement and closed questions identified the support for and against the scheme. A total of 55 respondents participated, including 18 AdPs, 25 CPs, 1 consultant trainee practitioner, 5 practitioners and 6 listed as 'other'. Forty-four participants were non-accredited, citing too much clinical work; no recognition from employers and too much effort for little reward. Motivations for joining the scheme were to improve the profession; help create a new consultant post and protect the non-clinical element of the consultant role. The CoR voluntary accreditation scheme has a small perceived value but overall, the majority of respondents believed the scheme did not warrant the work needed to apply. Concern was raised about the risk of creating a two-tier profession by the scheme's instigation. The results of this study suggest that the CoR's voluntary accreditation scheme would need to address these barriers before more practitioners would apply.


Getting Recovery Right After Neck Dissection (GRRAND-F): mixed-methods feasibility study to design a pragmatic randomised controlled trial protocol.
We will evaluate the feasibility of a randomised controlled trial to estimate the effectiveness and cost-effectiveness of a rehabilitation intervention on pain, function and health-related quality of life following neck dissection (ND) after head and neck cancer (HNC). This is a pragmatic, multicentred, feasibility study. Participants are randomised to usual care (control) or usual care plus an individualised, rehabilitation programme (Getting Recovery Right After Neck Dissection, GRRAND intervention). Adults aged over 18 with HNC for whom ND is part of their care will be recruited from specialist clinics. Participants are randomised in 1:1 ratio using a web-based service. The target sample size is 60 participants. Usual care will be received by all participants during their postoperative inpatient stay consisting standard National Health Service care supplemented with a booklet advising on postoperative self-management strategies. The GRRAND intervention programme consists of usual care plus up to six individual physiotherapy sessions including neck and shoulder range of motion (ROM) and progressive resistance exercises, advice and education. Between sessions participants will be advised to complete a home exercise programme. The primary outcome is to determine recruitment and retention rates from study participants across sites. Outcomes will be measured at 6 and 12 months. Participants and physiotherapists will be invited to an optional qualitative interview at the completion of their involvement in the study. The target qualitative sample size is 15 participants and 12 physiotherapists. Interviews aim to further investigate the feasibility and acceptability of the intervention and to determine wider experiences of the study design and intervention from patient and physiotherapist perspectives. Ethical approval was given on 29 October 2019 (National Research Ethics Committee Number: 19/SC/0457). Results will be reported at conferences and in peer-reviewed publications. ISRCTN11979997. Trial recruitment is ongoing and is expected to be completed by 30 August 2021.


Enhanced degradation of EGF receptors by a sorting nexin, SNX1.
The vectorial movement of proteins requires specific recognition by components of the vesicular trafficking machinery. A protein, sorting nexin-1 (SNX1), was identified in a human cell line that bound to a region of the epidermal growth factor receptor (EGFR) containing the lysosomal targeting code. SNX1 contains a region of homology to a yeast vacuolar sorting protein, and overexpression of SNX1 decreased the amount of EGFR on the cell surface as a result of enhanced rates of constitutive and ligand-induced degradation. Thus, SNX1 is likely to play a role in sorting EGFR to lysosomes.


Comparison of three cuffed emergency percutaneous cricothyroidotomy devices to conventional surgical cricothyroidotomy in a porcine model.
Emergency cricothyroidotomy is a potentially life-saving procedure in the 'cannot intubate cannot ventilate (CICV)' scenario. Although surgical cricothyroidotomy remains the technique recommended in many 'CICV' algorithms, the insertion of a tracheostomy as a cannula over a trocar, or using the Seldinger method, may have advantages as they are more familiar to the anaesthetist. We compared the utility of three cuffed cricothyroidotomy devices: cuffed Melker®, Quicktrach 2®, and PCK® devices, with surgical cricothyroidotomy. After ethical committee approval and written informed consent, 20 anaesthetists performed cricothyroidotomy with all four devices in random order, in a pig larynx and trachea model covered in cured pelt. The primary endpoints were the rate of successful placement of the cricothyroidotomy device into the trachea and the duration of the insertion attempt. The Melker® and Quicktrach 2® devices possessed advantages over the surgical approach, in contrast to the PCK® device, which performed less well. All 20 participants inserted the Melker®, with 19 being successful using the surgical approach and the Quicktrach 2®, whereas only 12 successfully inserted the PCK® device (PCK® vs surgical, P=0.02). The Quicktrach 2® had the fastest insertion times and caused least trauma to the posterior tracheal wall. The Melker® was rated highest by the participants and was the only device rated higher than the surgical technique. The Melker® and Quicktrach 2® devices appear to hold particular promise as alternatives to surgical cricothyroidotomy. Further studies, in more clinically relevant models, are required to confirm these initial positive findings.


Baculoviruses expressing the human familial Alzheimer's disease presenilin 1 mutation lacking exon 9 increase levels of an amyloid beta-like protein in Sf9 cells.
Presenilin 1 (PS1) plays a pivotal role in the production of the amyloid-beta protein (Abeta) that is central to the pathogenesis of Alzheimer's disease. PS1 regulates the intramembranous proteolysis of a 99-amino-acid C-terminal fragment of the amyloid precursor protein (APP-C99), a cleavage event that releases Abeta following a reaction catalyzed by an enzyme termed 'gamma-secretase'. The molecular mechanism of PS1-mediated, gamma-secretase cleavage remains largely unresolved. In particular, controversy surrounds whether PS1 includes the catalytic site of the gamma-secretase protease or whether instead PS1 mediates gamma-secretase activity indirectly, perhaps by regulating the trafficking or presentation of substrates to the 'authentic' protease, which may be a molecule distinct from PS1. To address this issue, the baculovirus expression system was used to co-express: (i) APP-C99; (ii) a pathogenic, constitutively active mutant form of PS1 lacking exon 9 (PS1DeltaE9); (iii) nicastrin and (iv) tropomyosin in Spodoptera frugiperda (Sf9) cells. Cells infected with APP-C99 alone produced an Abeta-like species, and levels of this species were enhanced by the addition of baculoviruses bearing the PS1DeltaE9 mutation. The addition to APP-C99-infected cells of baculoviruses bearing nicastrin, also a transmembrane protein, had a neutral or inhibitory effect on the reaction; tropomyosin viruses had the same effect as nicastrin viruses. These results suggest that PS1DeltaE9 molecules expressed in Sf9 cells retain the ability to modulate Abeta levels. Baculoviral-expressed PS1DeltaE9 provides a source of microgram quantities of bioactive molecules for use as starting material for purifying and reconstituting gamma-secretase activity from its individual purified component parts.


C21orf5, a novel human chromosome 21 gene, has a Caenorhabditis elegans ortholog (pad-1) required for embryonic patterning.
To contribute to the development of the transcription map of human chromosome 21 (HC21), we isolated a new transcript, C21orf5 (chromosome 21 open reading frame 5), encoding a predicted 2298-amino-acid protein. Analysis of the genomic DNA sequence revealed that C21orf5 consists of 37 exons that extend over 130 kb and maps between the CBR3 (carbonyl reductase 3) and the KIAA0136 genes. Northern blot analyses showed a ubiquitously expressed RNA species of 8.5 kb. RNA in situ hybridization on brain sections of normal human embryos revealed a strong labeling in restricted areas of the cerebral cortex. In silico analysis of the deduced C21orf5 protein revealed several highly probable transmembrane segments but no known protein domains or homology with known proteins. However, there were significant homologies to several hypothetical Caenorhabditis elegans proteins and Drosophila melanogaster genomic sequences. To investigate the function of C21orf5, we isolated the cDNA of the C. elegans ortholog and performed double-stranded RNA-mediated genetic interference experiments. The major phenotype observed in the progeny of injected animals was embryonic lethality. Most of the tissues of the embryo failed to undergo proper patterning during gastrulation, and morphogenesis did not occur; thus we termed the ortholog pad-1, for patterning defective 1. These results indicated that pad-1 is essential for the development and the survival of C. elegans. This study provides the first example of the use of C. elegans as a model to study the function of genes on human chromosome 21 that might be involved in Down syndrome.


Standardized representation, visualization and searchable repository of antiretroviral treatment-change episodes.
To identify the determinants of successful antiretroviral (ARV) therapy, researchers study the virological responses to treatment-change episodes (TCEs) accompanied by baseline plasma HIV-1 RNA levels, CD4+ T lymphocyte counts, and genotypic resistance data. Such studies, however, often differ in their inclusion and virological response criteria making direct comparisons of study results problematic. Moreover, the absence of a standard method for representing the data comprising a TCE makes it difficult to apply uniform criteria in the analysis of published studies of TCEs. To facilitate data sharing for TCE analyses, we developed an XML (Extensible Markup Language) Schema that represents the temporal relationship between plasma HIV-1 RNA levels, CD4 counts and genotypic drug resistance data surrounding an ARV treatment change. To demonstrate the adaptability of the TCE XML Schema to different clinical environments, we collaborate with four clinics to create a public repository of about 1,500 TCEs. Despite the nascent state of this TCE XML Repository, we were able to perform an analysis that generated a novel hypothesis pertaining to the optimal use of second-line therapies in resource-limited settings. We also developed an online program (TCE Finder) for searching the TCE XML Repository and another program (TCE Viewer) for generating a graphical depiction of a TCE from a TCE XML Schema document. The TCE Suite of applications - the XML Schema, Viewer, Finder, and Repository - addresses several major needs in the analysis of the predictors of virological response to ARV therapy. The TCE XML Schema and Viewer facilitate sharing data comprising a TCE. The TCE Repository, the only publicly available collection of TCEs, and the TCE Finder can be used for testing the predictive value of genotypic resistance interpretation systems and potentially for generating and testing novel hypotheses pertaining to the optimal use of salvage ARV therapy.


Treatment of adult short bowel syndrome with recombinant human growth hormone: a review of clinical studies.
Extensive resection of the intestinal tract frequently results in inadequate digestion and/or absorption of nutrients, a condition known as short bowel syndrome (SBS). Several therapies, including parenteral nutrition (PN), bowel rehabilitation, and surgical procedures to reconstruct the bowel, have been used for patients with SBS. However, these treatments only partially correct the underlying problem of reduced bowel function in some patients. This review investigates the results of studies of recombinant human growth hormone (r-hGH) in patients with SBS. In one randomized, controlled and five open-label studies, treatment with r-hGH significantly increased absorption of energy, protein and/or carbohydrates. Four studies also demonstrated significantly increased body weight, lean body mass, total body potassium and/or total body water in response to r-hGH. However, in two other randomized controlled trials, r-hGH had no significant effect on energy or fluid absorption. In one randomized, controlled study and six open-label trials, treatment with r-hGH also allowed a majority of patients, including those with no colon and significant bowel resection, to eliminate or substantially reduce PN. Optimal clinical benefits appear to be achieved when r-hGH is administered in combination with a specialized oral diet and perhaps with glutamine. Although the use of r-hGH to treat SBS patients remains controversial, results from these studies suggest that short-term r-hGH treatment in combination with optimized medical and dietetic treatments can enhance bowel absorption and function and, with the continuation of optimized medical and dietetic treatments, may reduce PN requirements for a wide range of patients with SBS.


Coupling of spin and angular momentum of light in plasmonic vortex.
We present that two distinct optical properties of light, the spin angular momentum (SAM) and the orbital angular momentum (OAM), can be coupled in the plasmonic vortex. If a plasmonic vortex lens (PVL) is illuminated by the helical vector beam (HVB) with the SAM and OAM, then those distinct angular momenta contribute to the generation of the plasmonic vortex together. The analytical model reveals that the total topological charge of the generated plasmonic vortex is given by a linear summation of those of the SAM and OAM, as well as the geometric charge of the PVL. The generation of the plasmonic vortex and the manipulation of the fractional topological charge are also presented.


Delayed S-cone sensitivity losses following the onset of intense yellow backgrounds linked to the lifetime of a photobleaching product?
Thirty years ago, Mollon, Stockman, & Polden (1987) reported that after the onset of intense yellow 581-nm backgrounds, S-cone threshold rose unexpectedly for several seconds before recovering to the light-adapted steady-state value-an effect they called: "transient-tritanopia of the second kind" (TT2). Given that 581-nm lights have little direct effect on S-cones, TT2 must arise indirectly from the backgrounds' effects on the L- and M-cones. We attribute the phenomenon to the action of an unknown L- and M-cone photobleaching product, X, which acts at their outputs like an "equivalent" background light that then inhibits S-cones at a cone-opponent, second-site. The time-course of TT2 is similar in form to the lifetime of X in a two-stage, first-order biochemical reaction A→X→C with successive best-fitting time-constants of 3.09 ± 0.35 and 7.73 ± 0.70 s. Alternatively, with an additional slowly recovering exponential "restoring-force" with a best-fitting time-constant 23.94 ± 1.42 s, the two-stage best-fitting time-constants become 4.15 ± 0.62 and 6.79 ± 1.00 s. Because the time-constants are roughly independent of the background illumination, and thus the rate of photoisomerization, A→X is likely to be a reaction subsidiary to the retinoid cycle, perhaps acting as a buffer when the bleaching rate is too high. X seems to be logarithmically related to S-cone threshold, which may result from the logarithmic cone-opponent, second-site response compression after multiplicative first-site adaptation. The restoring-force may be the same cone-opponent force that sets the rate of S-cone recovery following the unusual threshold increase following the offset of dimmer yellow backgrounds, an effect known as "transient-tritanopia" (TT1).


Plant metabolites. Structure and in vitro antiviral activity of quinovic acid glycosides from Uncaria tomentosa and Guettarda platypoda.
A reinvestigation of the bark of Uncaria tomentosa afforded, in addition to the major quinovic acid glycosides 1-3, three further glycosides 4-6. The structures were elucidated by spectral and chemical studies. Furthermore, a series of antiviral tests were performed on all these glycosides and on the related glycosides 7-9, previously isolated from Guettarda platypoda.


Neuroendoscopic approach to intraventricular lesions.
Object. The purpose of this study was to determine the efficacy of endoscopic treatment in patients with intraventricular tumors. Methods. A series of 30 patients with endoscopically treated intraventricular lesions is reported. The lesions included seven colloid cysts, six astrocytomas, three subependymomas, two ependymomas, and one each of the following: pineoblastoma, pineocytoma/pineoblastoma (intermediate type), epidermoid cyst, pineal cyst, medulloblastoma, arteriovenous hemangioma, cavernoma, choroid plexus papilloma, pituitary adenoma, craniopharyngioma, melanoma, and germinoma. Total tumor resections, partial resections, biopsies, stent implantations, septostomies, and third ventriculostomies were performed. In two cases (two subependymomas > 2 cm in diameter), piecemeal endoscopic resection was ineffective because of the very firm consistency of the tumors. Therefore the endoscopic procedure was discontinued and the tumors were removed microsurgically. In the remaining cases the procedures were completed as planned. Even in the presence of difficulties such as poor orientation or significant bleeding, there was no need to abandon the endoscopic procedure. A total of 28 strictly endoscopic interventions were performed, in which the average duration was 85 minutes (range 35-170 minutes). All colloid cysts and the epidermoid lesion were completely evacuated and the capsules were widely resected. Total extirpation of solid tumors was achieved in five cases, whereas most astrocytomas were partially resected. The hydrocephalus-related symptoms resolved in all of the 22 patients with cerebrospinal fluid pathway obstruction. There were no endoscopy-related deaths. In two cases, major bleeding occurred and was controlled endoscopically. The authors observed one case of meningitis, one of mutism, two of memory loss attributed to forniceal injury, one of transient trochlear palsy after a biopsy specimen of an aqueductal tumor was obtained, and one of transient confusion after a biopsy specimen of a germinoma was obtained. Conclusions. In the authors' preliminary experience, the endoscopic approach was found to be safe and effective. In this series, it was possible to achieve relief of noncommunicating hydrocephalus, tumor resections, and even complete tumor removals by using endoscopic techniques. Based on the results, the authors believe that endoscopic techniques should be considered in the treatment of selected intraventricular lesions.


Protein-binding elements establish in the oocyte the primary imprint of the Prader-Willi/Angelman syndromes domain.
Imprinting of the PWS/AS 2.4 Mb domain in the human is controlled by a paternally active imprinting center (PWS-IC). PWS-IC on the maternal allele is methylated and inactivated by an 880-bp sequence (AS-IC) located 30 kb upstream. In this communication, we report the identification of 7 cis acting elements within AS-IC. The elements: DMR, DNS, 2 OCTA sequences, SOX, E1, and E2 bind specific proteins that form at least 2 protein complexes. Using variants of an imprinted transgene, mutated at the elements each at a time, we show that (i) all 7 elements are involved in the methylation and inactivation of the maternal PWS-IC; (ii) the OCTA and SOX elements that bind a protein complex, and the E1 and E2 elements, function in establishing the primary imprint that constitutes an active and unmethylated AS-IC in the oocyte; (iii) DNS and DMR bind a multiprotein complex that may facilitate interaction between AS-IC and PWS-IC, mediating the inactivation in cis of PWS-IC; and (iv) all 7 elements participate in maintaining an unmethylated PWS-IC in the oocyte, which is essential for its maternal methylation later in development. Altogether, the above observations imply that the cis acting elements on AS-IC display diverse functions in establishing the imprints at both AS-IC and PWS-IC in the oocyte. A postulated epigenetic mark imprints the PWS-IC in the oocyte and maintains its inactive status during development before it is translated into maternal methylation.


New hemostatic agents in the combat setting.
Hemorrhage is a leading cause of potentially preventable death in both civilian and military trauma patients. Animal data have shown that hemostatic bandages reduce hemorrhage and improve survival. This article reports recent clinical observations regarding the efficacy and evolution of use of two new hemostatic bandages employed in the global war on terrorism. We performed a retrospective cohort review of soldiers treated with either the QuikClot or HemCon hemostatic bandages between April and October 2006. Hemostatic dressings were placed on wounds either in the field or at the combat support hospital (CSH). During the 6-month study period, 1691 trauma patients were admitted to the CSH. Fifty uses of hemostatic dressings in 44 patients (2.6% of admissions) were identified. Forty patients were treated with HemCon dressings, three patients with QuikClot, and one with both QuikClot and HemCon. Eighteen percent of the dressings were used in the field, predominantly on extremity wounds (7/8). In contrast, most dressings used in the CSH were for truncal wounds (26/36 patients). Hemostatic dressings were applied to extremity wounds in prehospital and hospital settings, either alone or in conjunction with tourniquets. In surviving patients (95%), the treating surgeon determined that the hemorrhage was either stopped or greatly decreased by use of hemostatic dressings. Two of the four patients treated with QuikClot had burns from exothermic reactions, while no adverse reactions were noted with HemCon. Hemostatic agents stop or decrease bleeding. Whereas HemCon appears to be safe, QuikClot may produce superficial burns. These new hemostatic agents have a place in the surgical armamentarium to assist in controlling internal hemorrhage from truncal and pelvic hemorrhage, especially during damage-control surgery.


Ovarian Epithelial Inclusions With Mucinous Differentiation: A Clinicopathologic Study of 42 Cases.
Ovarian epithelial inclusions lined by mucinous epithelium are rare and of uncertain origin. Ovaries containing such inclusions were studied in 42 women. The inclusions were divided into 3 groups: serous epithelial lined with typical ciliated morphology but with distinct basophilic cytoplasmic mucin in some or all of the lining cells, those lined by typical mucinous epithelium, and those lined by a combination of typical mucinous epithelium and serous epithelium. The mean patient age was 61.5 years. Pure mucinous inclusions were found in 27 patients, serous-type inclusions with cytoplasmic mucin in 20, and mixed type in 10. All 3 types of inclusions were found in 1 patient. Two types of inclusions were found in 13. Four patients had associated mucinous neoplasms (1 mucinous cystadenoma, 1 atypical proliferative seromucinous tumor, and 2 seromucinous cystadenomas), and 11 patients (26%) had endometriosis. The fallopian tubes in 4 patients (9.5%) also displayed mucinous metaplasia; this was not significantly different from the 3.1% we found in our previously reported series of unselected tubes from the same population. These findings suggest that mucinous inclusions may arise as a direct metaplastic change in serous-type inclusions. Other possible origins of mucinous inclusions in the ovarian cortex include endometriosis and Brenner (transitional cell) nests. Whether such inclusions can be a source of mucinous ovarian neoplasms as are Brenner tumors and mature cystic teratomas is unknown and may warrant further investigation.


Evidence for Linear but Not Helical Automatic Representation of Pitch in the Human Auditory System.
The perceptual organization of pitch is frequently described as helical, with a monotonic dimension of pitch height and a circular dimension of pitch chroma, accounting for the repeating structure of the octave. Although the neural representation of pitch height is widely studied, the way in which pitch chroma representation is manifested in neural activity is currently debated. We tested the automaticity of pitch chroma processing using the MMN-an ERP component indexing automatic detection of deviations from auditory regularity. Musicians trained to classify pure or complex tones across four octaves, based on chroma-C versus G (21 participants, Experiment 1) or C versus F# (27, Experiment 2). Next, they were passively exposed to MMN protocols designed to test automatic detection of height and chroma deviations. Finally, in an "attend chroma" block, participants had to detect the chroma deviants in a sequence similar to the passive MMN sequence. The chroma deviant tones were accurately detected in the training and the attend chroma parts both for pure and complex tones, with a slightly better performance for complex tones. However, in the passive blocks, a significant MMN was found only to height deviations and complex tone chroma deviations, but not to pure tone chroma deviations, even for perfect performers in the active tasks. These results indicate that, although height is represented preattentively, chroma is not. Processing the musical dimension of chroma may require higher cognitive processes, such as attention and working memory.


Skewed X-chromosome inactivation in scleroderma.
Scleroderma is a female-prevalent autoimmune disease of unclear etiology. Two fundamental gender differences, skewed X-chromosome inactivation (XCI) and pregnancy-related microchimerism, have been implicated in scleroderma. We investigated the XCI patterns of female scleroderma patients and the parental origin of the inactive X chromosome in those patients having skewed XCI patterns (>80%). In addition, we investigated whether a correlation exists between XCI patterns and microchimerism in a well-characterized cohort. About 195 female scleroderma patients and 160 female controls were analyzed for the androgen receptor locus to assess XCI patterns in the DNA extracted from peripheral blood cells. Skewed XCI was observed in 67 (44.9%) of 149 informative patients and in 10 of 124 healthy controls (8.0%) [odds ratio (OR) = 9.3 (95% confidence interval (CI) 4.3-20.6, P < 0.0001)]. Extremely skewed XCI (>90%) was present in 44 of 149 patients (29.5%) but only in 3 of 124 controls (2.4%; OR = 16.9; 95% CI 4.8-70.4, P < 0.0001). Parental origin of the inactive X chromosome was investigated for ten patients for whom maternal DNA was informative, and the inactive X chromosome was of maternal origin in eight patients and of paternal origin in two patients. Skewed XCI mosaicism could be considered as an important risk factor in scleroderma.


Malarial dihydroorotate dehydrogenase. Substrate and inhibitor specificity.
The malarial parasite relies on de novo pyrimidine biosynthesis to maintain its pyrimidine pools, and unlike the human host cell it is unable to scavenge preformed pyrimidines. Dihydroorotate dehydrogenase (DHODH) catalyzes the oxidation of dihydroorotate (DHO) to produce orotate, a key step in pyrimidine biosynthesis. The enzyme is located in the outer membrane of the mitochondria of the malarial parasite. To characterize the biochemical properties of the malarial enzyme, an N-terminally truncated version of P. falciparum DHODH has been expressed as a soluble, active enzyme in E. coli. The recombinant enzyme binds 0.9 molar equivalents of the cofactor FMN and it has a pH maximum of 8.0 (k(cat) 8 s(-1), K(m)(app) DHO (40-80 microm)). The substrate specificity of the ubiquinone cofactor (CoQ(n)) that is required for the oxidation of FMN in the second step of the reaction was also determined. The isoprenoid (n) length of CoQ(n) was a determinant of reaction efficiency; CoQ(4), CoQ(6) and decylubiquinone (CoQ(D)) were efficiently utilized in the reaction, however cofactors lacking an isoprenoid tail (CoQ(0) and vitamin K(3)) showed decreased catalytic efficiency resulting from a 4 to 7-fold increase in K(m)(app). Five potent inhibitors of mammalian DHODH, Redoxal, dichloroallyl lawsone (DCL), and three analogs of A77 1726 were tested as inhibitors of the malarial enzyme. All five compounds were poor inhibitors of the malarial enzyme, with IC(50)'s ranging from 0.1-1.0 mm. The IC(50) values for inhibition of the malarial enzyme are 10(2)-10(4)-fold higher than the values reported for the mammalian enzyme, demonstrating that inhibitor binding to DHODH is species specific. These studies provide direct evidence that the malarial DHODH active site is different from the host enzyme, and that it is an attractive target for the development of new anti-malarial agents.


Sex-Specific Association of Apolipoprotein E With Cerebrospinal Fluid Levels of Tau.
The strongest genetic risk factor for Alzheimer disease (AD), the apolipoprotein E (APOE) gene, has a stronger association among women compared with men. Yet limited work has evaluated the association between APOE alleles and markers of AD neuropathology in a sex-specific manner. To evaluate sex differences in the association between APOE and markers of AD neuropathology measured in cerebrospinal fluid (CSF) during life or in brain tissue at autopsy. This multicohort study selected data from 10 longitudinal cohort studies of normal aging and AD. Cohorts had variable recruitment criteria and follow-up intervals and included population-based and clinic-based samples. Inclusion in our analysis required APOE genotype data and either CSF data available for analysis. Analyses began on November 6, 2017, and were completed on December 20, 2017. Biomarker analyses included levels of β-amyloid 42, total tau, and phosphorylated tau measured in CSF. Autopsy analyses included Consortium to Establish a Registry for Alzheimer's Disease staging for neuritic plaques and Braak staging for neurofibrillary tangles. Of the 1798 patients in the CSF biomarker cohort, 862 were women, 226 had AD, 1690 were white, and the mean (SD) age was 70 [9] years. Of the 5109 patients in the autopsy cohort, 2813 were women, 4953 were white, and the mean (SD) age was 84 (9) years. After correcting for multiple comparisons using the Bonferroni procedure, we observed a statistically significant interaction between APOE-ε4 and sex on CSF total tau (β = 0.41; 95% CI, 0.27-0.55; P < .001) and phosphorylated tau (β = 0.24; 95% CI, 0.09-0.38; P = .001), whereby APOE showed a stronger association among women compared with men. Post hoc analyses suggested this sex difference was present in amyloid-positive individuals (β = 0.41; 95% CI, 0.20-0.62; P < .001) but not among amyloid-negative individuals (β = 0.06; 95% CI, -0.18 to 0.31; P = .62). We did not observe sex differences in the association between APOE and β-amyloid 42, neuritic plaque burden, or neurofibrillary tangle burden. We provide robust evidence of a stronger association between APOE-ε4 and CSF tau levels among women compared with men across multiple independent data sets. Interestingly, APOE-ε4 is not differentially associated with autopsy measures of neurofibrillary tangles. Together, the sex difference in the association between APOE and CSF measures of tau and the lack of a sex difference in the association with neurofibrillary tangles at autopsy suggest that APOE may modulate risk for neurodegeneration in a sex-specific manner, particularly in the presence of amyloidosis.


mTNF reverse signalling induced by TNFα antagonists involves a GDF-1 dependent pathway: implications for Crohn's disease.
Mechanisms of action (MoA) of anti-tumour necrosis factor α (TNFα) therapies in Crohn's disease (CD) may critically involve induction of immune cell apoptosis via membrane-bound TNFα (mTNFα) binding. Certolizumab pegol (CZP), which is effective in induction and maintenance of remission in CD lacks the ability to induce apoptosis. The aim of this study was to analyse transcriptomal responses of reverse signalling induced by the TNFα binding agents infliximab (IFX) and CZP in myelomonocytic cells. Induction of transcriptional patterns upon anti-TNFα stimulation was assessed using oligonucleotide microarrays. mRNA expression of GDF-1/ LASS1, which was identified as a shared target, was studied in inflammatory bowel disease by real-time PCR, while signalling pathways induced by growth and differentiation factor 1 (GDF-1) were investigated using western blots and ELISA. IFX and CZP induced a common signature of 20 transcripts that could be categorised into control of cell cycle, transcription activation and pre-mRNA processing. We selected GDF-1/LASS1 for functional follow-up, which was found to be upregulated in inflamed CD tissues. We show that downregulation of GDF-1/LASS1 depends on autocrine release of transforming growth factor β after mTNFα ligation. We demonstrate that GDF-1 itself acts as a novel proinflammatory factor via induction of interleukin 6 and signal transducer and activator of transcription 3 and is downregulated after IFX treatment. Commonalities in the MoA of IFX and CZP comprise modulation of non-apoptotic pathways through downregulation of proinflammatory GDF-1. Further characterisation of the molecular role of GDF-1 in complex inflammatory processes in vivo is warranted to decide whether this proinflammatory molecule is a promising therapeutic target in patients with CD.