Prevalence of Salmonella in juvenile dogs affected with parvoviral enteritis.
Salmonellosis is a disease of major zoonotic importance and canine parvovirus is a potentially fatal cause of canine enteritis with a world-wide distribution. Persistent isolation of Salmonella during routine environmental sampling surveys of a hospital ward, reserved for the treatment of dogs with canine parvovirus infection, prompted investigation into a possible source. We hypothesised that dogs affected by canine parvovirus would have a higher prevalence of faecal salmonellae compared to an apparently healthy cohort. Seventy-four client-owned dogs naturally infected with canine parvovirus and 42 apparently healthy client-owned dogs were included in the study. This prospective, longitudinal, observational study was conducted over an 18-month period. Fresh faecal samples were collected from dogs aged 6 weeks to 9 months diagnosed with canine parvovirus infection and admitted for treatment, and from apparently healthy dogs presented for vaccination or routine hospital procedures. Faeces were submitted for the isolation, antimicrobial susceptibility testing and serotyping of salmonellae. The prevalence of faecal Salmonella shedding was 22% and 31% for the affected and apparently healthy dogs, respectively, which was not statistically different. No significant associations between Salmonella status and possible risk factors or continuous variables such as age, body weight and duration of hospitalisation were identified. All the Salmonella isolates (n = 32) were resistant to penicillin G, lincomycin and tylosin. Salmonellae from nine different serotypes were identified. The prevalence of Salmonella shedding in both groups was higher than that commonly reported, yet similar to those in previous reports on young dogs, shelter dogs or dogs fed a raw meat diet.


Berberine Directly Affects the Gut Microbiota to Promote Intestinal Farnesoid X Receptor Activation.
Intestinal bacteria play an important role in bile acid metabolism and in the regulation of multiple host metabolic pathways (e.g., lipid and glucose homeostasis) through modulation of intestinal farnesoid X receptor (FXR) activity. Here, we examined the effect of berberine (BBR), a natural plant alkaloid, on intestinal bacteria using in vitro and in vivo models. In vivo, the metabolomic response and changes in mouse intestinal bacterial communities treated with BBR (100 mg/kg) for 5 days were assessed using NMR- and mass spectrometry-based metabolomics coupled with multivariate data analysis. Short-term BBR exposure altered intestinal bacteria by reducing Clostridium cluster XIVa and IV and their bile salt hydrolase (BSH) activity, which resulted in the accumulation of taurocholic acid (TCA). The accumulation of TCA was associated with activation of intestinal FXR, which can mediate bile acid, lipid, and glucose metabolism. In vitro, isolated mouse cecal bacteria were incubated with three doses of BBR (0.1, 1, and 10 mg/ml) for 4 hours in an anaerobic chamber. NMR-based metabolomics combined with flow cytometry was used to evaluate the direct physiologic and metabolic effect of BBR on the bacteria. In vitro, BBR exposure not only altered bacterial physiology but also changed bacterial community composition and function, especially reducing BSH-expressing bacteria like Clostridium spp. These data suggest that BBR directly affects bacteria to alter bile acid metabolism and activate FXR signaling. These data provide new insights into the link between intestinal bacteria, nuclear receptor signaling, and xenobiotics.


Therapeutic Implications of a Specific Murine Monoclonal Antibody (7E3) to the Platelet Receptor GPIIb/IIIa.
The murine monoclonal antibody 7E3 blocks the platelet glycoprotein IIb/IIIa receptor, and is a potent inhibitor of platelet function in both animals and man. Animal models of the acute coronary syndromes suggest that 7E3 abolishes the in vivo formation of platelet thrombi, accelerates thrombolysis with tissue plasminogen activator, and prevents subsequent reocclusion. Human studies with 7E3 suggest that complete inhibition of platelet function may be safely undertaken for periods of up to 36 h, and preliminary studies indicate effectiveness in the therapy of clinical unstable angina. Potential problems with its use include immunogenicity and thrombocytopenia. The outcome of the acute coronary thrombotic syndromes, which include unstable angina, acute myocardial infarction and abrupt closure following coronary angioplasty, may be significantly improved with 7E3 therapy.


Arthroscopic extended rotator interval release for treating refractory adhesive capsulitis.
To present the clinical results of arthroscopic extended rotator interval release with a stretching program for treating refractory adhesive capsulitis. Case series; level of evidence, 4. Arthroscopy-assisted extended rotator interval tissue release including anterior capsular was performed in 26 patients with refractory adhesive capsulitis. All rotator interval tissues, except the medial sling of the biceps, were excised and the excursion of the subscapularis tendon was restored and freely mobilized. The preoperative mean passive forward flexion was 101°, whereas external rotation at the side was 10°. Patients were followed for a minimum of 2 years and their visual analog scale for pain, muscle power, range of motion, Constant score, modified American Shoulder and Elbow Surgeons Shoulder Evaluation Form score, and modified University of California at Los Angeles score were recorded. Visual pain scale and the aforementioned clinical scores improved postoperatively. The patients exhibited a significant postoperative difference in forward flexion, external rotation, and internal rotation. Postoperative mean passive forward flexion was 172°, whereas external rotation at the side was 58°. There was no difference in the muscle power postoperatively including abduction, internal rotation, and external rotation. Our study revealed satisfactory subjective and objective clinical results after a 2-year follow-up. Arthroscopy-assisted extended rotator interval release with a stretching program could be an alternative treatment for refractory adhesive capsulitis.


Risk of hemodialysis graft thrombosis: analysis of monthly flow surveillance.
During clinical application of flow surveillance of hemodialysis grafts, the risk of thrombosis is assessed month after month, rather than after one or several measurements, as has been done in published studies. Adequate assessment of risk should consider the many measurements obtained over time. Prospective cohort diagnostic test study. 176 patients with hemodialysis grafts from 2 university-affiliated dialysis units during a 6-year period. Monthly measurement of graft blood flow or change in flow. Graft thrombosis. We used logistic regression analysis to compute the risk of thrombosis and used receiver operating characteristic (ROC) curves to assess the accuracy in predicting thrombosis within 1 month. Newer grafts were most likely to thrombose, whereas older grafts were unlikely to thrombose even at low flows or large decreases in flow. Areas under the ROC curves were 0.698 for flow and 0.713 for change in flow measured over 2 months. Flow predicted thrombosis with a sensitivity of 53% at a specificity of 79%, and change in flow had a sensitivity of 58% at a specificity of 75%. More than half the thromboses lacked a change in flow measurement, usually because thrombosis occurred before a change could be measured. Thus, the effective predictive accuracy of change in flow was much less than the ROC curves indicated because the curves do not consider missing measurements. Performance characteristics of index tests may vary across patient populations. Flow and change in flow are inaccurate predictors of thrombosis. Many thromboses are not predicted, and intervention based on surveillance likely yields many unnecessary procedures. Thus, this study does not support routine application of surveillance to prevent thrombosis.


Neuropathological consequences of delivering an adenoviral vector in the rat brain.
Adenoviruses have many advantages as vehicles for gene delivery to the central nervous system (CNS) and retrograde transport of vectors to axonally linked sites has been postulated as a method for targeting neurons in remote brain regions. To investigate optimisation of this we injected different doses of vector and have documented the neuropathological side effects. Increasing doses of a first-generation adenoviral vector, expressing the lacZ gene, were inoculated in the rat striatum and beta-galactosidase expression was examined at the primary and secondary sites. Subsequently, at the highest dose of vector, transgene expression, the inflammatory response, tyrosine hydroxylase (TH) expression and the rotational behaviour of animals were studied over time. When a high dose of an adenoviral vector was delivered to the rat striatum, high levels of transgene expression were seen at 5 days in the injection site and in the substantia nigra. Smaller doses gave lower levels of expression with little expression detectable in the substantia nigra. At later time points, with the high dose, a marked reduction in transgene expression was detected and was accompanied by cytopathic damage, a strong inflammatory response and animal weight loss. This was associated with depletion in TH levels and abnormal motor behaviour in animals. Neuropathological damage in the dopaminergic system, caused by high doses of adenoviral vectors, has not previously been documented. To minimise damage and prolong transgene expression, it is important that the dose of vectors to be delivered is carefully optimised.


Trans-endocytosis elicited by nectins transfers cytoplasmic cargo, including infectious material, between cells.
Here, we show that cells expressing the adherens junction protein nectin-1 capture nectin-4-containing membranes from the surface of adjacent cells in a trans-endocytosis process. We find that internalized nectin-1-nectin-4 complexes follow the endocytic pathway. The nectin-1 cytoplasmic tail controls transfer: its deletion prevents trans-endocytosis, while its exchange with the nectin-4 tail reverses transfer direction. Nectin-1-expressing cells acquire dye-labeled cytoplasmic proteins synchronously with nectin-4, a process most active during cell adhesion. Some cytoplasmic cargo remains functional after transfer, as demonstrated with encapsidated genomes of measles virus (MeV). This virus uses nectin-4, but not nectin-1, as a receptor. Epithelial cells expressing nectin-4, but not those expressing another MeV receptor in its place, can transfer infection to nectin-1-expressing primary neurons. Thus, this newly discovered process can move cytoplasmic cargo, including infectious material, from epithelial cells to neurons. We name the process nectin-elicited cytoplasm transfer (NECT). NECT-related trans-endocytosis processes may be exploited by pathogens to extend tropism. This article has an associated First Person interview with the first author of the paper.


Toward 'omic scale metabolite profiling: a dual separation-mass spectrometry approach for coverage of lipid and central carbon metabolism.
Although the objective of any 'omic science is broad measurement of its constituents, such coverage has been challenging in metabolomics because the metabolome is comprised of a chemically diverse set of small molecules with variable physical properties. While extensive studies have been performed to identify metabolite isolation and separation methods, these strategies introduce bias toward lipophilic or water-soluble metabolites depending on whether reversed-phase (RP) or hydrophilic interaction liquid chromatography (HILIC) is used, respectively. Here we extend our consideration of metabolome isolation and separation procedures to integrate RPLC/MS and HILIC/MS profiling. An aminopropyl-based HILIC/MS method was optimized on the basis of mobile-phase additives and pH, followed by evaluation of reproducibility. When applied to the untargeted study of perturbed bacterial metabolomes, the HILIC method enabled the accurate assessment of key, dysregulated metabolites in central carbon pathways (e.g., amino acids, organic acids, phosphorylated sugars, energy currency metabolites), which could not be retained by RPLC. To demonstrate the value of the integrative approach, bacterial cells, human plasma, and cancer cells were analyzed by combined RPLC/HILIC separation coupled to ESI positive/negative MS detection. The combined approach resulted in the observation of metabolites associated with lipid and central carbon metabolism from a single biological extract, using 80% organic solvent (ACN:MeOH:H2O 2:2:1). It enabled the detection of more than 30,000 features from each sample type, with the highest number of uniquely detected features by RPLC in ESI positive mode and by HILIC in ESI negative mode. Therefore, we conclude that when time and sample are limited, the maximum amount of biological information related to lipid and central carbon metabolism can be acquired by combining RPLC ESI positive and HILIC ESI negative mode analysis.


Microwave accelerated Glaser-Hay macrocyclizations at high concentrations.
Efficient macrocyclization can be conducted at high concentrations employing microwave irradiation and a phase separation strategy. The rate of the Glaser-Hay macrocyclization is accelerated using microwave irradiation and reaction times decreased from 48 h to 1-6 h, depending on the nature of the substrate. Macrocyclization concentrations could be increased up to 0.1 M compared to traditional concentrations (0.2 mM).


An experimental study on the noise properties of x-ray CT sinogram data in Radon space.
Computed tomography (CT) has been well established as a diagnostic tool through hardware optimization and sophisticated data calibration. For screening purposes, the associated x-ray exposure risk must be minimized. An effective way to minimize the risk is to deliver fewer x-rays to the subject or lower the mAs parameter in data acquisition. This will increase the data noise. This work aims to study the noise property of the calibrated or preprocessed sinogram data in Radon space as the mAs level decreases. An anthropomorphic torso phantom was scanned repeatedly by a commercial CT imager at five different mAs levels from 100 down to 17 (the lowest value provided by the scanner). The preprocessed sinogram datasets were extracted from the CT scanner to a laboratory computer for noise analysis. The repeated measurements at each mAs level were used to test the normality of the repeatedly measured samples for each data channel using the Shapiro-Wilk statistical test merit. We further studied the probability distribution of the repeated measures. Most importantly, we validated a theoretical relationship between the sample mean and variance at each channel. It is our intention that the statistical test and particularly the relationship between the first and second statistical moments will improve low-dose CT image reconstruction for screening applications.


Spatiotemporal Analysis of Malaria in Urban Ahmedabad (Gujarat), India: Identification of Hot Spots and Risk Factors for Targeted Intervention.
The world population, especially in developing countries, has experienced a rapid progression of urbanization over the last half century. Urbanization has been accompanied by a rise in cases of urban infectious diseases, such as malaria. The complexity and heterogeneity of the urban environment has made study of specific urban centers vital for urban malaria control programs, whereas more generalizable risk factor identification also remains essential. Ahmedabad city, India, is a large urban center located in the state of Gujarat, which has experienced a significant Plasmodium vivax and Plasmodium falciparum disease burden. Therefore, a targeted analysis of malaria in Ahmedabad city was undertaken to identify spatiotemporal patterns of malaria, risk factors, and methods of predicting future malaria cases. Malaria incidence in Ahmedabad city was found to be spatially heterogeneous, but temporally stable, with high spatial correlation between species. Because of this stability, a prediction method utilizing historic cases from prior years and seasons was used successfully to predict which areas of Ahmedabad city would experience the highest malaria burden and could be used to prospectively target interventions. Finally, spatial analysis showed that normalized difference vegetation index, proximity to water sources, and location within Ahmedabad city relative to the dense urban core were the best predictors of malaria incidence. Because of the heterogeneity of urban environments and urban malaria itself, the study of specific large urban centers is vital to assist in allocating resources and informing future urban planning.


Health-related quality of life for patients with vasculitis and their spouses.
Chronic disease affects both patients and spouses, yet the effect of vasculitis on the spouses of patients has not been systematically examined. We addressed this research gap by describing 8 dimensions of health-related quality of life (HRQOL) for antineutrophil cytoplasmic antibody-associated vasculitis (AAV) patients and their spouses and compared patients and spouses with norms for the general US population. We also determined whether the impact of AAV on HRQOL differs by sex for patients or their spouses. Ninety-seven patients with AAV and their spouses completed a mailed questionnaire that included demographic information and a measure of HRQOL (Medical Outcomes Study Short Form 36.) Using the RAND method, we calculated 8 HRQOL dimensions: general health, physical functioning, emotional role limitations, physical role limitations, social functioning, mental health, bodily pain, and energy/vitality. Using norm-based scores, we compared the HRQOL of patients and spouses with the general US population. We then used multivariate analysis of covariance (MANCOVA) test for sex differences in HRQOL for patients and spouses. Patients with AAV scored lower than the US norm on all HRQOL subscales with the exception of bodily pain. In contrast, spouses scored similarly to national norms. When age, education, race, illness duration, and disease severity were controlled, there were no significant sex differences in HRQOL for patients or spouses. AAV negatively impacts HRQOL for patients; whereas, spouses do not appear to be as negatively impacted. Long-term marriages may attenuate the effect of sex on HRQOL. Quality of life should be assessed throughout the disease course for both patients and spouses.


Protein Expression in Tonsillar and Base of Tongue Cancer and in Relation to Human Papillomavirus (HPV) and Clinical Outcome.
Human papillomavirus (HPV) is a major etiological factor for tonsillar and the base of tongue cancer (TSCC/BOTSCC). HPV-positive and HPV-negative TSCC/BOTSCC present major differences in mutations, mRNA expression and clinical outcome. Earlier protein studies on TSCC/BOTSCC have mainly analyzed individual proteins. Here, the aim was to compare a larger set of cancer and immune related proteins in HPV-positive and HPV-negative TSCC/BOTSCC in relation to normal tissue, presence of HPV, and clinical outcome. Fresh frozen tissue from 42 HPV-positive and 17 HPV-negative TSCC/BOTSCC, and corresponding normal samples, were analyzed for expression of 167 proteins using two Olink multiplex immunoassays. Major differences in protein expression between TSCC/BOTSCC and normal tissue were identified, especially in chemo- and cytokines. Moreover, 34 proteins, mainly immunoregulatory proteins and chemokines, were differently expressed in HPV-positive vs HPV-negative TSCC/BOTSCC. Several proteins were potentially related to clinical outcome for HPV-positive or HPV-negative tumors. For HPV-positive tumors, these were mostly related to angiogenesis and hypoxia. Correlation with clinical outcome of one of these, VEGFA, was validated by immunohistochemistry. Differences in immune related proteins between HPV-positive and HPV-negative TSCC/BOTSCC reflect the stronger activity of the immune defense in the former. Angiogenesis related proteins might serve as potential targets for therapy in HPV-positive TSCC/BOTSCC.


Phenotype ontologies and cross-species analysis for translational research.
The use of model organisms as tools for the investigation of human genetic variation has significantly and rapidly advanced our understanding of the aetiologies underlying hereditary traits. However, while equivalences in the DNA sequence of two species may be readily inferred through evolutionary models, the identification of equivalence in the phenotypic consequences resulting from comparable genetic variation is far from straightforward, limiting the value of the modelling paradigm. In this review, we provide an overview of the emerging statistical and computational approaches to objectively identify phenotypic equivalence between human and model organisms with examples from the vertebrate models, mouse and zebrafish. Firstly, we discuss enrichment approaches, which deem the most frequent phenotype among the orthologues of a set of genes associated with a common human phenotype as the orthologous phenotype, or phenolog, in the model species. Secondly, we introduce and discuss computational reasoning approaches to identify phenotypic equivalences made possible through the development of intra- and interspecies ontologies. Finally, we consider the particular challenges involved in modelling neuropsychiatric disorders, which illustrate many of the remaining difficulties in developing comprehensive and unequivocal interspecies phenotype mappings.


[Laser pointers are not toys; eye injury with permanent loss of visual acuity].
In the nineteen-nineties, there was much hype in the European media about presumed laser pointer maculopathy. However, the recent introduction of more powerful and therefore more dangerous laser pointers and their easy availability on the internet necessitates vigilance on the issue. This is an urgent matter, as here we report three cases of proven maculopathy due to an unsafe laser pointer. Three boys aged 13, 9 and 12 years used an unsafe laser pointer as a toy and looked repeatedly into the pointer, resulting in a permanent reduction in visual acuity due to macular damage. Laser pointers are not designed to be children's toys or instruments to annoy people in a crowd. Health authorities and the ophthalmic community should be aware of the potential danger of improper use of high-output laser pointers and warn the general public before the widespread availability of unsafe laser pointers and consequently laser pointer-induced macular damage becomes a true social problem.


Behavioral reduction of infection risk.
Evolutionary biologists have long postulated that there should be fitness advantages to animals that are able to recognize and avoid conspecifics infected with contact-transmitted disease. This avoidance hypothesis is in direct conflict with much of epidemiological theory, which is founded on the assumptions that the likelihood of infection is equal among members of a population and constant over space. The inconsistency between epidemiological theory and the avoidance hypothesis has received relatively little attention because, to date, there has been no evidence that animals can recognize and reduce infection risk from conspecifics. We investigated the effects of Candida humicola, a pathogen that reduces growth rates and can cause death of tadpoles, on associations between infected and uninfected individuals. Here we demonstrate that bullfrog (Rana catesbeiana) tadpoles avoid infected conspecifics because proximity influences infection. This avoidance behavior is stimulated by chemical cues from infected individuals and thus does not require direct contact between individuals. Such facultative modulations of disease infection risk may have critical consequences for the population dynamics of disease organisms and their impact on host populations.


Invasive plants facilitated by socioeconomic change harbor vectors of scrub typhus and spotted fever.
Ecological determinants of most emerging vector-borne diseases are understudied, particularly for neglected tropical disease. Moreover, although socioeconomic impacts can have significant downstream effects on human risks to vector-borne diseases via a change in land cover, particularly facilitating the invasion of exotic plants, related studies remains very scarce. Scrub typhus and spotted fever are neglected diseases emerging around the globe and are transmitted by chigger mites and ticks infective of Orientia tsutsugamushi and Rickettsia spp., respectively, with small mammals as the primary hosts of both vectors. We investigated how invasions of the plant Leucaena leucocephala caused by widespread abandonment of farmlands driven by industrialization affected abundance of chiggers and ticks in Penghu Island, Taiwan. We determined ectoparasite abundance by trapping small mammals in three types of habitats (invasion site, agricultural field, human residential) every two months for a year. Based on ectoparasite burdens, invasion sites harbored more chiggers and ticks than the other two habitats. Furthermore, hosts maintained higher burdens of both vectors in early winter and burdens of chiggers were more stable across seasons in invasion sites, suggesting that sites with invasive plants could be a temporary refuge for both vectors and might help mitigate the negative influence of unfavorable climate. Infective rates of O. tsutsugamushi in chiggers and Rickettsia in ticks were also consistently not lower in invasion sites. Top soil temperature and relative humidity were similar across the three habitats, but invasion sites contained more of the rat Rattus losea, on which chiggers and ticks were more engorged than those from the most commonly trapped species (Suncus murinus shrew), indicating that abundance of the host R. losea instead of microclimate might better determine the abundance of both vectors. This study highlights an important but largely neglected issue that socioeconomic change can have unexpected consequences for human health induced particularly by invasive plants, which could become a hotspot for emerging infectious diseases but usually are very hard to be eradicated. In the future, a more comprehensive approach that integrates socio-economics, land use, exotic species, and human health should be considered to fully understand potential emergence of vector-borne diseases.


Contributions to Speech-Cue Weighting in Older Adults With Impaired Hearing.
Purpose In a previous paper (Souza, Wright, Blackburn, Tatman, & Gallun, 2015), we explored the extent to which individuals with sensorineural hearing loss used different cues for speech identification when multiple cues were available. Specifically, some listeners placed the greatest weight on spectral cues (spectral shape and/or formant transition), whereas others relied on the temporal envelope. In the current study, we aimed to determine whether listeners who relied on temporal envelope did so because they were unable to discriminate the formant information at a level sufficient to use it for identification and the extent to which a brief discrimination test could predict cue weighting patterns. Method Participants were 30 older adults with bilateral sensorineural hearing loss. The first task was to label synthetic speech tokens based on the combined percept of temporal envelope rise time and formant transitions. An individual profile was derived from linear discriminant analysis of the identification responses. The second task was to discriminate differences in either temporal envelope rise time or formant transitions. The third task was to discriminate spectrotemporal modulation in a nonspeech stimulus. Results All listeners were able to discriminate temporal envelope rise time at levels sufficient for the identification task. There was wide variability in the ability to discriminate formant transitions, and that ability predicted approximately one third of the variance in the identification task. There was no relationship between performance in the identification task and either amount of hearing loss or ability to discriminate nonspeech spectrotemporal modulation. Conclusions The data suggest that listeners who rely to a greater extent on temporal cues lack the ability to discriminate fine-grained spectral information. The fact that the amount of hearing loss was not associated with the cue profile underscores the need to characterize individual abilities in a more nuanced way than can be captured by the pure-tone audiogram.


Selective optical-optical switching for planar plasmonic waveguides and nodes.
Two nodes are studied as means to perform selective optical-optical switching of four planar thin surface plasmon waveguides by interfering TEM(10), TEM(01), and TEM(00) light beams incident upon a node. One node uses a flat-apex pyramidal reflector to reflect the incident light toward the waveguides' ends. An alternative node is a simple square aperture, which couples surface plasmons through light diffraction at the aperture's edges. Numerical calculations predict switching contrast, coupling efficiencies, and cross-talk between waveguides. Individually turned-off waveguides are shown to have their coupled surface plasmons attenuated by at least -10 dB and up to -21 dB.


High Rate of Positive Anti-Tissue Transglutaminase Antibodies in Chronic Liver Disease.
Since the recognition of tissue transglutaminase (tTG) as the target antigen of anti-endomysium antibodies, several ELISA assays using either guinea pig or human recombinant tTG have been developed. The aim of the study was to compare the behaviour of anti-tTG and anti-endomysium antibodies assays in coeliacs and in patients with chronic liver disease. 34 patients (24 women, 34.9 ± 12.5 years) with coeliac disease and 41 with chronic liver disease (14 women, 57 ± 11.2 years), including 19 cirrhotics, were evaluated for anti-endomysium antibodies by indirect immunofluorescence and for anti-tTG IgA antibodies by ELISA, using guinea pig liver or human recombinant transglutaminase. The prevalences of anti-tTG and anti-endomysium antibodies were 100% in patients with coeliac disease at diagnosis, 75% and 64.3% in patients on a gluten-free diet. All liver disease patients were negative for anti-endomysium antibodies, while 11 (26.8%) were positive for anti-tTG. All these patients had liver cirrhosis and represented 57.9% of all cirrhotics. The presence of anti-tTG was associated with higher Child-Pugh scores. The use of human transglutaminase determined a reduction in the rate of positive results; however, the rate of positive anti-tTG was still 17.1% in all liver disease patients and 31.6% in cirrhotics. Our data confirm that anti-tTG have a similar sensitivity compared with anti-endomysium antibodies assay in coeliacs. However, a high prevalence of positive anti-tTG results is observed in cirrhotic patients, even when human recombinant tTG is used. The high prevalence of positive results among cirrhotic patients is associated with more advanced liver disease.


MGMT Promoter Methylation Is a Strong Prognostic Biomarker for Benefit from Dose-Intensified Temozolomide Rechallenge in Progressive Glioblastoma: The DIRECTOR Trial.
Rechallenge with temozolomide (TMZ) at first progression of glioblastoma after temozolomide chemoradiotherapy (TMZ/RT→TMZ) has been studied in retrospective and single-arm prospective studies, applying temozolomide continuously or using 7/14 or 21/28 days schedules. The DIRECTOR trial sought to show superiority of the 7/14 regimen. Patients with glioblastoma at first progression after TMZ/RT→TMZ and at least two maintenance temozolomide cycles were randomized to Arm A [one week on (120 mg/m(2) per day)/one week off] or Arm B [3 weeks on (80 mg/m(2) per day)/one week off]. The primary endpoint was median time-to-treatment failure (TTF) defined as progression, premature temozolomide discontinuation for toxicity, or death from any cause. O(6)-methylguanine DNA methyltransferase (MGMT) promoter methylation was prospectively assessed by methylation-specific PCR. Because of withdrawal of support, the trial was prematurely closed to accrual after 105 patients. There was a similar outcome in both arms for median TTF [A: 1.8 months; 95% confidence intervals (CI), 1.8-3.2 vs. B: 2.0 months; 95% CI, 1.8-3.5] and overall survival [A: 9.8 months (95% CI, 6.7-13.0) vs. B: 10.6 months (95% CI, 8.1-11.6)]. Median TTF in patients with MGMT-methylated tumors was 3.2 months (95% CI, 1.8-7.4) versus 1.8 months (95% CI, 1.8-2) in MGMT-unmethylated glioblastoma. Progression-free survival rates at 6 months (PFS-6) were 39.7% with versus 6.9% without MGMT promoter methylation. Temozolomide rechallenge is a treatment option for MGMT promoter-methylated recurrent glioblastoma. Alternative strategies need to be considered for patients with progressive glioblastoma without MGMT promoter methylation.


Caspase-8 gene therapy using the human telomerase reverse transcriptase promoter for malignant glioma cells.
Telomerase is a distinctive candidate for targeted gene therapy of malignant gliomas, because the vast majority of malignant gliomas express telomerase activity while normal brain tissues do not. Recently, we developed a telomerase-specific expression system of caspase-8 gene using the promoter of the human telomerase reverse transcriptase (hTERT) gene. However, the transcriptional activity of hTERT-181 promoter (a 181-base pair [bp] region upstream of the transcription start site) was relatively lower in malignant glioma cells than in other tumors such as prostate cancer cells. To establish the hTERT/caspase-8 construct as a novel therapy for malignant gliomas, we need to increase the transcriptional activity of the hTERT promoter in malignant glioma cells. In the present study, we demonstrate that the transcriptional activity of hTERT-378 promoter (a 378-bp region) was 2- to 40-fold higher in hTERT-positive malignant glioma cells (A172, GB-1, T98G, U87-MG, U251-MG, and U373-MG) than that of hTERT-181. We further demonstrate that by using the hTERT-378/caspase-8 construct, apoptosis was restricted to malignant glioma cells, and was not seen in astrocytes or fibroblasts lacking hTERT. Moreover, the growth of subcutaneously established U373-MG tumors in mice was significantly inhibited by seven daily intratumoral injections of hTERT-378/caspase-8 construct and its inhibitory effect persisted during 3 additional weeks without additional treatment. These results suggest that the telomerase-specific expression of caspase-8 under hTERT-378 promoter is a novel targeting approach for the treatment of telomerase-positive malignant gliomas.


An Overview of Specialist Services for Huntington's Disease in the United Kingdom.
Huntington's disease (HD) is a rare inherited neurodegenerative disorder characterized by complex evolving needs that change as the condition progresses. There is limited understanding about the organization of HD clinical services and their resourcing in the United Kingdom (UK). To understand the organization and resourcing of specialist HD services for people with HD (PwHD) in the UKMethods:This cross-sectional study collected quantitative data via on online survey, and qualitative data via telephone semi-structured interviews. Descriptive statistics were used to describe quantitative outcomes, and qualitative results were analyzed using content analysis. A total of 31 specialist services for HD were identified. Of the 27 services that completed the online survey, 23 had an active multidisciplinary team of healthcare professionals (HCPs) and were led primarily by a mental health trust (26%) or tertiary referral hospital (26%). Specialist services offered outpatient clinics (96%), outreach in the community (74%), telemedicine (70%), inpatient beds (26%) and satellite clinics (26%). Many services indicated that their capacity (ability to see patients as often as needed with current resources) was difficult, with some services reporting more difficulty at the early or later stages of HD. Key resourcing gaps were identified with access to facilities, HCPs and referral networks. This research highlights the variation in organization and capacity within individual HD services as well as current resourcing and gaps in access that influence this capacity. Further research should be done to understand the impact of service organization and current resourcing gaps in access on the quality of care provided for PwHD in the UK.


Neonatal survival interventions in humanitarian emergencies: a survey of current practices and programs.
Neonatal deaths account for over 40% of all deaths in children younger than five years of age and neonatal mortality rates are highest in areas affected by humanitarian emergencies. Of the ten countries with the highest neonatal mortality rates globally, six are currently or recently affected by a humanitarian emergency. Yet, little is known about newborn care in crisis settings. Understanding current policies and practices for the care of newborns used by humanitarian aid organizations will inform efforts to improve care in these challenging settings. Between August 18 and September 25, 2009, 56 respondents that work in humanitarian emergencies completed a web-based survey either in English or French. A snow ball sampling technique was used to identify organizations that provide health services during humanitarian emergencies to gather information on current practices for maternal and newborn care in these settings. Information was collected about continuum-of-care services for maternal, newborn and child health, referral services, training and capacity development, health information systems, policies and guidelines, and organizational priorities. Data were entered into MS Excel and frequencies and percentages were calculated. The majority of responding organizations reported implementing components of neonatal and maternal health interventions. However, multiple barriers exist in providing comprehensive care, including: funding shortages (63.3%), gaps in training (51.0%) and staff shortages and turnover (44.9%). Neonatal care is provided by most of the responding humanitarian organizations; however, the quality, breadth and consistency of this care are limited.


Clustering huge protein sequence sets in linear time.
Metagenomic datasets contain billions of protein sequences that could greatly enhance large-scale functional annotation and structure prediction. Utilizing this enormous resource would require reducing its redundancy by similarity clustering. However, clustering hundreds of millions of sequences is impractical using current algorithms because their runtimes scale as the input set size N times the number of clusters K, which is typically of similar order as N, resulting in runtimes that increase almost quadratically with N. We developed Linclust, the first clustering algorithm whose runtime scales as N, independent of K. It can also cluster datasets several times larger than the available main memory. We cluster 1.6 billion metagenomic sequence fragments in 10 h on a single server to 50% sequence identity, >1000 times faster than has been possible before. Linclust will help to unlock the great wealth contained in metagenomic and genomic sequence databases.


Normalising comparative effectiveness trials as clinical practice.
There is a lack of high-quality evidence underpinning many contemporary clinical practice guidelines embedded in the healthcare systems, leading to treatment uncertainty and practice variation in most medical disciplines. Comparative effectiveness trials (CETs) represent a diverse range of research that focuses on optimising health outcomes by comparing currently approved interventions to generate high-quality evidence to inform decision makers. Yet, despite their ability to produce real-world evidence that addresses the key priorities of patients and health systems, many implementation challenges exist within the healthcare environment.This manuscript aims to highlight common barriers to conducting CETs and describes potential solutions to normalise their conduct as part of a learning healthcare system.


Numerical modeling of nitrogen removal processes in biofilters with simultaneous nitritation and anammox.
This study developed a simple numerical model for nitrogen removal in biofilters, which was designed to enhance simultaneous nitritation and anaerobic ammonium oxidation (anammox). It is the first attempt to simulate anammox together with two-step nitrification in natural treatment systems, which may have different kinetic parameters and temperature effects from conventional bioreactors. Prediction accuracy was improved by adjusting kinetic coefficients over the startup period of the biofilters. The maximum rates of nitritation and nitrite oxidation increased linearly over time during the startup period. Simulations confirmed successful enhancement of simultaneous nitritation and anammox (SNA) in the biofilters, with anammox contributing 35% of ammonium removal. Effluent ammonium concentration was affected by influent ammonium concentration and the maximum nitritation rate, and was insensitive to the maximum nitrite oxidation rate and anammox substrate factor. Ammonium removal via SNA was likely limited by biomass of aerobic ammonia oxidizing bacteria in the biofilters. The developed model is a promising tool for studying the dynamics of nitrogen removal processes including SNA in natural treatment systems.


Prehospital experiences of older men with a first myocardial infarction: a qualitative analysis within the Northern Sweden MONICA Study.
To explore older men's prehospital experiences of their first myocardial infarction (MI). The delay between the onset of symptoms to the initiation of medical care is a major determinant of prognosis in MI. The majority of people experiencing MI are men. But few studies have been conducted solely on men's experiences before seeking medical care for MI. The objective of this study was to explore older men's experiences of symptoms and their reasoning during the prehospital phase of their first MI. Data collection was carried out through individual interviews with 20 men representing the age range 65-80 (mean 71) years. The participants were interviewed 3 days after admission for a confirmed first MI. The interviews were designed to prompt the men to describe their symptoms and their reasoning up to the decision to seek care. A qualitative content analysis was used to analyse the interviews. The prehospital phase in older men was found to be a complex and extended journey. The symptoms were perceived from diffuse ill-being to a cluster of alarming symptoms. The participants dealt with conceptions about MI symptoms. They were unsure about the cause of their symptoms, which did not correspond to their expectations about an MI, and whether they should seek medical care. They had difficulty making the final decision to seek care and strived to maintain a normal life. They initially tried to understand, reduce or treat the symptoms by themselves. The decision to seek medical care preceded a movement from uncertainty to conviction. Understanding older men's prehospital experiences of MI is essential to reduce their patient decision times. This requires knowledge about the complexity and dynamic evolvement of symptoms, beliefs and strategies to maintain an ordinary life.


Branched polyethylenimine-functionalized carbon dots as sensitive and selective fluorescent probes for N-acetylcysteine via an off-on mechanism.
N-Acetylcysteine (NAC) plays an important role in optimizing the protective ability of cells as well as in the treatment of some chronic clinical conditions. Unfortunately, current methods for determining NAC based on fluorescence assay strategies remain poorly investigated. In this study, a new fluorescence method for highly sensitive and selective detection of NAC was developed. The detection method employed the fluorescence (FL) signal of branched polyethylenimine-functionalized carbon dots (PEI-CDs) via an off-on mechanism. The detection system was based on the formation of cupric amine complexes by the reaction of Cu2+ ions with surface amino groups on PEI-CDs. The FL of PEI-CDs at 460.0 nm upon exciting at 360.0 nm was quenched as a result of electron transfer between the complexes and PEI-CDs. Upon addition of NAC to Cu2+-CD solution, electron transfer occurred from the mercapto group on NAC to Cu2+, leading to the formation of Cu+ species and the dissociation of cupric amine complexes. As a result, the FL signal of PEI-CDs was turned on since single excited electrons cannot be transferred from PEI-CDs to the cupric amine complexes. The detection limit of this method was 0.56 μM, while the linear response ranged from 5.56 μM to 277.8 μM.


Effect of implant surface and grafting on implants placed into simulated extraction sockets: a histologic study in dogs.
To study bone healing at implant sites in simulated extraction sockets with 1-mm marginal defects and compare healing around a turned surface (T) to that around a porous oxide surface prepared by anodic oxidation (AO) with or without the use of an autogenous bone graft. All mandibular premolars and first molars were extracted from 10 mongrel dogs. After 9 weeks, four sites were prepared on both sides of all mandibles. Each osteotomy was widened in the coronal 5 mm to create a marginal defect of 1 mm around the implants. Autogenous bone was collected during the drilling procedure. The sites were randomized to receive implants with a T or an AO surface, with or without bone grafting. The animals were sacrificed 4 months after implant placement for histologic analysis. Clinically, all sites healed with complete bone fill. The combination of an AO implant and a bone graft resulted in a significantly greater percentage of bone-to-implant contact (BIC) (P < .05) versus all other groups. The highest point of BIC was achieved with the AO group, which was significantly greater than the lowest group (T). No significant differences between groups were found when the apical 4 mm (non-gap areas) were compared (P = .65). Studies have demonstrated that bone can fill in a marginal defect around a titanium implant with varied histologic BIC, depending on implant surface type and defect dimensions. Based upon this animal study using 10 mongrel dogs, marginal circumferential defects of 1 mm showed significantly higher BIC values for implants that were prepared by AO compared to implants with a turned surface. The addition of autogenous bone grafts further enhanced the degree of BIC.


Ribosomal protein S1 and NusA protein complexed to recombination protein beta of phage lambda.
The red genes of bacteriophage lambda specify two proteins, exonuclease and beta protein, which are essential for general recombination of lambda in recA cells. Earlier studies suggested that these proteins form an equimolar complex (C. M. Radding, J. Rosenweig, F. Richards, and E. Cassuto, J. Biol. Chem. 246:2510-2512, 1971). A more recent study indicated that beta protein forms a strong complex with an unknown polypeptide of 70 kDa (K. Muniyappa and C. M. Radding, J. Biol. Chem. 261:7472-7478, 1986). In the present study, in addition to the complex of beta and the 70-kDa protein, a new association of beta protein with a 65-kDa protein was observed. N-terminal sequencing identified these proteins as host-encoded ribosomal protein S1 and transcription terminator protein NusA.


An amplitude quotient based method to analyze changes in the shape of the glottal pulse in the regulation of vocal intensity.
This study presents an approach to visualizing intensity regulation in speech. The method expresses a voice sample in a two-dimensional space using amplitude-domain values extracted from the glottal flow estimated by inverse filtering. The two-dimensional presentation is obtained by expressing a time-domain measure of the glottal pulse, the amplitude quotient (AQ), as a function of the negative peak amplitude of the flow derivative (d(peak)). The regulation of vocal intensity was analyzed with the proposed method from voices varying from extremely soft to very loud with a SPL range of approximately 55 dB. When vocal intensity was increased, the speech samples first showed a rapidly decreasing trend as expressed on the proposed AQ-d(peak) graph. When intensity was further raised, the location of the samples converged toward a horizontal line, the asymptote of a hypothetical hyperbola. This behavior of the AQ-d(peak) graph indicates that the intensity regulation strategy changes from laryngeal to respiratory mechanisms and the method chosen makes it possible to quantify how control mechanisms underlying the regulation of vocal intensity change gradually between the two means. The proposed presentation constitutes an easy-to-implement method to visualize the function of voice production in intensity regulation because the only information needed is the glottal flow wave form estimated by inverse filtering the acoustic speech pressure signal.


Evolving DNA methylation and gene expression markers of B-cell chronic lymphocytic leukemia are present in pre-diagnostic blood samples more than 10 years prior to diagnosis.
B-cell chronic lymphocytic leukemia (CLL) is a common type of adult leukemia. It often follows an indolent course and is preceded by monoclonal B-cell lymphocytosis, an asymptomatic condition, however it is not known what causes subjects with this condition to progress to CLL. Hence the discovery of prediagnostic markers has the potential to improve the identification of subjects likely to develop CLL and may also provide insights into the pathogenesis of the disease of potential clinical relevance. We employed peripheral blood buffy coats of 347 apparently healthy subjects, of whom 28 were diagnosed with CLL 2.0-15.7 years after enrollment, to derive for the first time genome-wide DNA methylation, as well as gene and miRNA expression, profiles associated with the risk of future disease. After adjustment for white blood cell composition, we identified 722 differentially methylated CpG sites and 15 differentially expressed genes (Bonferroni-corrected p < 0.05) as well as 2 miRNAs (FDR < 0.05) which were associated with the risk of future CLL. The majority of these signals have also been observed in clinical CLL, suggesting the presence in prediagnostic blood of CLL-like cells. Future CLL cases who, at enrollment, had a relatively low B-cell fraction (<10%), and were therefore less likely to have been suffering from undiagnosed CLL or a precursor condition, showed profiles involving smaller numbers of the same differential signals with intensities, after adjusting for B-cell content, generally smaller than those observed in the full set of cases. A similar picture was obtained when the differential profiles of cases with time-to-diagnosis above the overall median period of 7.4 years were compared with those with shorted time-to-disease. Differentially methylated genes of major functional significance include numerous genes that encode for transcription factors, especially members of the homeobox family, while differentially expressed genes include, among others, multiple genes related to WNT signaling as well as the miRNAs miR-150-5p and miR-155-5p. Our findings demonstrate the presence in prediagnostic blood of future CLL patients, more than 10 years before diagnosis, of CLL-like cells which evolve as preclinical disease progresses, and point to early molecular alterations with a pathogenetic potential.


Transcription rate and transcript length drive formation of chromosomal interaction domain boundaries.
Chromosomes in all organisms are highly organized and divided into multiple chromosomal interaction domains, or topological domains. Regions of active, high transcription help establish and maintain domain boundaries, but precisely how this occurs remains unclear. Here, using fluorescence microscopy and chromosome conformation capture in conjunction with deep sequencing (Hi-C), we show that in Caulobacter crescentus, both transcription rate and transcript length, independent of concurrent translation, drive the formation of domain boundaries. We find that long, highly expressed genes do not form topological boundaries simply through the inhibition of supercoil diffusion. Instead, our results support a model in which long, active regions of transcription drive local decompaction of the chromosome, with these more open regions of the chromosome forming spatial gaps in vivo that diminish contacts between DNA in neighboring domains. These insights into the molecular forces responsible for domain formation in Caulobacter likely generalize to other bacteria and possibly eukaryotes.


Natural products modulate Shigella-host-cell interaction.
This study focused on identifying possible new options derived from natural sources for the treatment of bacterial infections. Several natural products were investigated for their potential in modulating Shigella-host-cell interactions. The proliferation of Shigella sonnei was effectively inhibited inside HEp-2 cells in the presence of 4-methoxycinnamic acid and propolin D. Propolin D also significantly reduced the apoptosis of infected macrophage-like U937 cells and moderately reduced the secretion of interleukin (IL)-1β and IL-18, which probably resulted from the inhibition of invasion plasmid antigen B secretion by this compound. Further characterization showed that propolin D did not prevent escape of Shigella from phagocytic vacuoles, as evidenced by actin-based motility and by the fact that addition of chloroquine did not further reduce the number of intracellular c.f.u. The role of propolin D in modulating autophagy could not be established under the experimental conditions used. As these compounds had no direct anti-Shigella activity in vitro, it was concluded that these compounds modulated Shigella-host-cell interactions by targeting yet-to-be defined mechanisms that provide benefits to host cells.


Lymphoma--emerging realities in sub-Saharan Africa.
Substantial geographical differences exist for Hodgkin and other lymphoproliferative disorders with these having previously been documented in a report from the lymphoma reclassification project. In the light of rampant human immunodeficiency syndrome, largely centred in sub-Sahara, this experience is updated in a further 512 consecutive individuals treated over an 8-year period in a privately based academic centre. Median age was 55.2 years 61% were males, 10% had Hodgkin lymphoma and, overall, constitutional symptoms were present in 20%. Prior to referral 19% had received chemotherapy and a further 20% some form of irradiation. Median survival in hairy cell leukaemia (n=14), chronic lymphocytic leukaemia-small lymphocytic lymphoma (n=103), Hodgkin (n=41) and follicular lymphoma (n=59) was not reached at the time of analysis and exceeded 36 months. This was followed by 32 months for those with mantle cell (n=7), splenic (n=2) and extranodal marginal cell (n=11), 24 months for T-cell lymphomas (n=24), 20 months for diffuse large B-cell variants (n=88) but only 12 months for the aggressive tumours exemplified by Burkitt (n=7) and lymphoblastic subtypes (n=6). The remaining 36 patients had to be excluded because numbers were too small for statistical analysis or unreliable staging. Adverse factors were constitutional symptoms, prior treatment with chemotherapy, intermediate or high-risk scores as defined by the international prognostic index, histologic grading and certain anatomical sites of primary tumour. In contrast gender, staging by Rye or Rai classification, retroviral infection and prior treatment with radiotherapy were without effect. Overall survival at 3 years in each category was compared to the curve for the entire cohort and was 100% in hairy cell leukaemia receiving two chlorodeoxyadenosine and greater than 88% in Hodgkin lymphoma treated according to the German study group protocols (p=0.0004). Corresponding figures for chronic lymphocytic leukaemia-small lymphocytic lymphoma were 82% (p=0.0006), follicular lymphoma 71% (p=0.060), peripheral T-cell lymphoma 43% (p=0.0156), diffuse large B-cell lymphoma 39% (p<0.0001), aggressive tumours 25% (p=0.0002) and for the indolent categories including mantle cell, splenic and extra nodal marginal cell lymphomas 22% (p=0.2023). Outcome argues in favour of patient management by a multidisciplinary team implicit in which are standardised protocols for diagnosis, staging and treatment. Under these circumstances the well recognized centre effect applies when results approximate those from first world reference centres. Conversely any deviation from such a disciplined approach is unlikely to achieve comparable benefit and therefore to be strongly discouraged.


Unusual kinetic isotope effects of deuterium reinforced polyunsaturated fatty acids in tocopherol-mediated free radical chain oxidations.
Substitution of -CD2- at the reactive centers of linoleic and linolenic acids reduces the rate of abstraction of D by a tocopheryl radical by as much as 36-fold, compared to the abstraction of H from a corresponding -CH2- center. This H atom transfer reaction is the rate-determining step in the tocopherol-mediated peroxidation of lipids in human low-density lipoproteins, a process that has been linked to coronary artery disease. The unanticipated large kinetic isotope effects reported here for the tocopherol-mediated oxidation of linoleic and linolenic acids and esters suggests that tunneling makes this process favorable.


Visual and non-visual properties of filters manipulating short-wavelength light.
Optical filters and tints manipulating short-wavelength light (sometimes called 'blue-blocking' or 'blue-attenuating' filters) are used in the management of a range of ocular, retinal, neurological and psychiatric disorders. In many cases, the only available quantification of the optical effects of a given optical filter is the spectral transmittance, which specifies the amount of light transmitted as a function of wavelength. We propose a novel physiologically relevant and retinally referenced framework for quantifying the visual and non-visual effects of these filters, incorporating the attenuation of luminance (luminous transmittance), the attenuation of melanopsin activation (melanopsin transmittance), the colour shift, and the reduction of the colour gamut (gamut reduction). Using these criteria, we examined a novel database of spectral transmittance functions of optical filters (n = 121) which were digitally extracted from a variety of sources. We find a large diversity in the alteration of visual and non-visual properties. The spectral transmittance properties of the examined filters vary widely, in terms of shapes and cut-off wavelengths. All filters show relatively more melanopsin attenuation than luminance attenuation (lower melanopsin transmittance than luminous transmittance). Across the data set, we find that melanopsin transmittance and luminous transmittance are correlated. We suggest that future studies and examinations of the physiological effects of optical filters quantify the visual and non-visual effects of the filters beyond the spectral transmittance, which will eventually aid in developing a mechanistic understanding of how different filters affect physiology. We strongly discourage comparing the downstream effects of different filters on, e.g. sleep or circadian responses, without considering their effects on the retinal stimulus.


The prevalence and significance of a short QT interval in 18,825 low-risk individuals including athletes.
The short QT syndrome is a cardiac channelopathy characterised by accelerated repolarisation which manifests as a short QT interval on the ECG. The definition of a short QT interval is debated, ranging from <390 to ≤320 ms, and its clinical significance in healthy young individuals is unknown. We assessed the prevalence and medium-term significance of an isolated short QT interval in a diverse young British population. Between 2005 and 2013, 18 825 apparently healthy people aged 14-35 years underwent cardiovascular evaluation with history, physical examination and ECG. QT intervals were measured by cardiologists using 4 recommended guidelines (Seattle 2013, Heart Rhythm Society 2013, European Society of Cardiology 2010 and American Heart Association 2009). The prevalence of a short QT interval was 0.1% (26 patients, ≤320 ms), 0.2% (44 patients, ≤330 ms), 7.9% (1478 patients, <380 ms), 15.8% (2973 patients, <390 ms). Male gender and Afro-Caribbean ethnicity had the strongest association with short QT intervals. Athletes had shorter QT intervals than non-athletes but athletic status did not predict short QT intervals. Individuals with short QT intervals ≤320 ms did not report syncope or a sinister family history, and during a follow-up period of 5.3±1.2 years, there were no deaths in this group. The prevalence of a short QT interval depends on the recommended cut-off value. Even at values ≤320 ms, there was an excellent medium-term prognosis among 14 people followed. We conclude that a definition of ≤320 ms is realistic to prevent overdiagnosis and excessive investigations.


Reducing the dimensionality of data with neural networks.
High-dimensional data can be converted to low-dimensional codes by training a multilayer neural network with a small central layer to reconstruct high-dimensional input vectors. Gradient descent can be used for fine-tuning the weights in such "autoencoder" networks, but this works well only if the initial weights are close to a good solution. We describe an effective way of initializing the weights that allows deep autoencoder networks to learn low-dimensional codes that work much better than principal components analysis as a tool to reduce the dimensionality of data.


A criss-cross heart with concordant atrioventriculo-arterial connections. Report of a case.
A distinctive angiographic appearance is described in a case of "criss-cross" heart with concordant atrioventriculo-arterial connections. The atrial situs was normal, and the morphological right ventricle was superior to the left ventricle, and the ventricular septum was horizontal in position. Both ventricleswere connected by a small ventricular septal defect. The aorta was situated anteriorly and to the right of the pulmonary artery. Although the cardiac segment was the apparent [S,L,D] type, the heart had conconrdant atrioventriculo-arterial connections. The ventricular rotation of the solitus heart about the longitudinal and the anteroposterior axis affected the atrioventricular flows, the plane of the ventricular septum, the inflow and outflow tracts of the ventricles, the interrelationship of the great arteries, and the course of the left coronary artery. Using the recent definition of criss-cross heart, we classified the heart as solitus-concordant (l-rotated) -normal.


Matrix analyses of interaction among fingers in static force production tasks.
The fingers on a hand show interactions in force production tasks. The interfinger connection matrices (IFMs) quantify these interactions (Li et al. 2002; Zatsiorsky et al. 2002b; Danion et al. 2003). The goal of the present study was to explore the differences in the IFMs of individual subjects and, in particular, to establish a procedure that may be used in the future for diagnostic purposes. Subjects ( n=20) pressed downward maximally with ten different combinations of the four fingers, index (I), middle (M), ring (R), and little (L): I, M, R, L, IM, MR, RL, IMR, MRL, and IMRL. Voluntary activation of a subset of the four fingers was accompanied by an involuntary force production by fingers that were not intentionally activated (enslaving). Interfinger connection matrices were computed for each subject by the artificial neural network. The similarities/dissimilarities (proximities) between the individual matrices were determined. This procedure was performed twice: (a) for nonnormalized IFMs whose elements represented the amount of force (in newtons) exerted by a finger i in response to a unit command to a finger j; and (b) for normalized IFMs, after dividing the elements of each IFM by the total force produced by the four fingers acting together (the elements of the matrix are in percents). The 20x20 matrix of the proximities was subjected to multidimensional scaling (MDS) to reduce the number of dimensions and identify the major ones. To interpret the meaning of the computed dimensions, they were regressed on a set of finger force parameters described in the text. For the nonnormalized IFMs an interpretable dimension was the strength of the subjects. For the normalized IFMs two dimensions were interpreted: (a) the location of the point of resultant force application along the mediolateral axis that is defined by the pattern of force sharing among the fingers and (b) the total contribution of the enslaved forces into the total finger force. We speculate that the similarity of typical everyday tasks across the population promotes the similarity of the IMFs reflecting optimal hand functioning over these tasks.


Enteric Pathogens in HIV/AIDS from a Tertiary Care Hospital.
Patterns of enteric infections in HIV in developing countries may differ in several important ways from developed countries, the knowledge of which can often guide therapy when resource limitations hamper the exact diagnosis of the etiological agent in HIV-associated diarrhea. The primary objective of this study was to define and compare the microbial etiologies of diarrhea in HIV-1 infected and non infected patients and in HIV infected non diarrheal patients. This study was conducted between April 2007 and July 2007 at the Department of Microbiology, Maulana Azad Medical College, New Delhi. Stool samples from 50 HIV seropositive cases with diarrhea (study group), 50 HIV seropositive cases without diarrhea (control group I), and 50 HIV seronegative cases with diarrhea (control group II) were examined. After the diagnosis of HIV infection was made, routine parasitological and bacteriological detection was done. An ELISA was used for the detection of Clostridium difficile toxin and Cryptosporidium antigen in stool samples. The overall prevalence of enteric parasitosis in the study group was 20% and the bacteria identified were Escherischia coli in 24% of the case, Clostridium difficile in 10% of the cases, Salmonella species and Vibrio cholerae in 4% of the cases, and Shigella species in 2% of the cases. Candida species was identified in 36% of the cases. Identification of the etiological agent of diarrhea in a patient with AIDS is very important as it can help in the institution of appropriate therapy and the reduction of morbidity and mortality in these patients.


Domain structure for an amorphous iridium-oxide water-oxidation catalyst characterized by X-ray pair distribution function analysis.
The domain structure of an amorphous, "blue layer" iridium-oxide water-oxidation catalyst film (BL) electrodeposited from the soluble precursor complex, [Cp*Ir(H2O)3]SO4, was characterized by X-ray pair distribution function (PDF) analysis. The results show that the experimental PDF can be fit remarkably well using a single Ir5O22 cluster extracted from the rutile lattice. The model includes distortions that indicate the presence of Ir(μ-O)3Ir or distorted Ir(μ-O)2Ir substructures, and hence deviations from a rutile structure. The five Ir atom cluster is suggested to represent the population-averaged distribution of metal-oxo clusters in the film. BL is found to be distinguished from other amorphous film water-oxidation catalysts because of the remarkably small domain size and homogeneity. As such, the blue layer catalyst provides a model for investigating ligand-determined metal-oxide cluster assembly and catalyst mechanism.


A novel bile acid biosynthesis defect due to a deficiency of peroxisomal ABCD3.
ABCD3 is one of three ATP-binding cassette (ABC) transporters present in the peroxisomal membrane catalyzing ATP-dependent transport of substrates for metabolic pathways localized in peroxisomes. So far, the precise function of ABCD3 is not known. Here, we report the identification of the first patient with a defect of ABCD3. The patient presented with hepatosplenomegaly and severe liver disease and showed a striking accumulation of peroxisomal C27-bile acid intermediates in plasma. Investigation of peroxisomal parameters in skin fibroblasts revealed a reduced number of enlarged import-competent peroxisomes. Peroxisomal beta-oxidation of C26:0 was normal, but beta-oxidation of pristanic acid was reduced. Genetic analysis revealed a homozygous deletion at the DNA level of 1758bp, predicted to result in a truncated ABCD3 protein lacking the C-terminal 24 amino acids (p.Y635NfsX1). Liver disease progressed and the patient required liver transplantation at 4 years of age but expired shortly after transplantation. To corroborate our findings in the patient, we studied a previously generated Abcd3 knockout mouse model. Abcd3-/- mice accumulated the branched chain fatty acid phytanic acid after phytol loading. In addition, analysis of bile acids revealed a reduction of C24 bile acids, whereas C27-bile acid intermediates were significantly increased in liver, bile and intestine of Abcd3-/- mice. Thus, both in the patient and in Abcd3-/- mice, there was evidence of a bile acid biosynthesis defect. In conclusion, our studies show that ABCD3 is involved in transport of branched-chain fatty acids and C27 bile acids into the peroxisome and that this is a crucial step in bile acid biosynthesis.


ApoE2 Exaggerates PTSD-Related Behavioral, Cognitive, and Neuroendocrine Alterations.
Apolipoprotein E (apoE) is an essential component of lipoprotein particles in both the brain and periphery, and exists in three isoforms in the human population: E2, E3, and E4. ApoE has numerous, well-established roles in neurobiology. Most notably, E4 is associated with earlier onset and increased risk of Alzheimer's disease (AD). Although possession of E2 is protective in the context of AD, E2 appears to confer an increased incidence and severity of posttraumatic stress disorder (PTSD). However, the biological processes underlying this link remain unclear. In this study, we began to elucidate these associations by examining the effects of apoE on PTSD severity in combat veterans, and on PTSD-like behavior in mice with human apoE. In a group of 92 veterans with PTSD, we observed significantly higher Clinician-Administered PTSD Scale and PTSD Checklist scores in E2+ individuals, as well as alterations in salivary cortisol levels. Furthermore, we measured behavioral and biological outcomes in mice expressing human apoE after a single stressful event as well as following a period of chronic variable stress, a model of combat-related trauma. Mice with E2 showed impairments in fear extinction, and behavioral, cognitive, and neuroendocrine alterations following trauma. To the best of our knowledge, these data constitute the first translational demonstration of PTSD severity in men and PTSD-like symptoms in mice with E2, and point to apoE as a novel biomarker of susceptibility, and potential therapeutic target, for PTSD.


[Relation of serum levels of C-reactive protein to anthropometric meaurements; a sustematic review of studies in South America].
In many studies the Body Mass Index (BMI) is associated with serum markers of inflammation such as CRP. In obesity there is an increase of adipose tissue resulting in an increase in BMI. It has been reported that adipose tissue has a high production and secretion of a variety of proinflammatory molecules such as tumor necrosis factor-α, interleukin-6, interleukin-8 and C-reactive protein, which may have local effects on the physiology of fat cell and systemic effects on other organs. To conduct a systematic review of studies conducted in South America where they relate serum levels of CRP and anthropometric measurements, BMI and waist circumference (CC) in the healthy adult population. We searched the PubMed database for articles published until January 2012, in English, Spanish and Portuguese, which relates serum levels of CRP and anthropometric measurements, BMI and WC. Search for words used PCR, BMI and WC and the names of each of the South American countries (Argentina, Bolivia, Brazil, Chile, Colombia, Ecuador, Guyana, Paraguay, Peru, Suriname, Uruguay and Venezuela). We identified 141 potential studies, of which 8 met the inclusion criteria. In studies serum levels of CRP were positively correlated with BMI (r 0.08 to 0.84) and CC (r 0.27 to 1.03), being highly relevant correlation between BMI and CC with CRP serum levels observed in obese people in South America. Serum levels of CRP, and therefore subclinical inflammation seems to be related to increased rates of anthropometric measures in the South American population. A better understanding of the mechanisms and molecular components of the inflammatory response induced by an increase in BMI may lead to identifying new therapeutic targets that can prevent the complications associated with obesity.


Do stage of disease, comorbidity or access to treatment explain socioeconomic differences in survival after ovarian cancer? - A cohort study among Danish women diagnosed 2005-2010.
In order to reduce social inequality in cancer survival, knowledge is needed about where in the cancer trajectory disparities occur, and how social and health-related aspects may interact. We aimed to determine whether socioeconomic factors are related to cancer diagnosis stage, and whether socioeconomic disparities in survival after ovarian cancer can be explained by socioeconomic differences in cancer stage, comorbidity, treatment or lifestyle factors. In the Danish Gynaecological Cancer Database we identified 2873 cases of ovarian cancer diagnosed between 2005 and 2010. From this data we retrieved information on prognostic factors, treatment information and lifestyle factors. Age, vital status, comorbidity, education, income and cohabitation status were ascertained from nationwide administrative registers. Associations were analyzed with logistic regression and Cox regression models. Educational level was weakly associated with cancer stage. Short education, lower income and living without a partner were related to poorer survival after ovarian cancer. Among women with early cancer stage, HR (95% CI) for death was 1.75 (1.20-2.54) in shorter compared to longer educated women. After adjustment for comorbid conditions, cancer stage, tumour histology, operation status and lifestyle factors, socioeconomic differences in survival persisted. Socioeconomic disparities in survival after ovarian cancer were to some extent, but not fully explained by differences in important prognostic factors, suggesting further investigations into this problem, however implying that socially less advantaged ovarian cancer patients should receive attention during cancer treatment and rehabilitation.


Studying weak and dynamic interactions of posttranslationally modified proteins using expressed protein ligation.
Many cellular processes are regulated by posttranslational modifications that are recognized by specific domains in protein binding partners. These interactions are often weak, thus allowing a highly dynamic and combinatorial regulatory network of protein-protein interactions. We report an efficient strategy that overcomes challenges in structural analysis of such a weak transient interaction between the Tudor domain of the Survival of Motor Neuron (SMN) protein and symmetrically dimethylated arginine (sDMA). The posttranslational modification is chemically introduced and covalently linked to the effector module by a one-pot expressed protein ligation (EPL) procedure also enabling segmental incorporation of NMR-active isotopes for structural analysis. Covalent coupling of the two interacting moieties shifts the equilibrium to the bound state, and stoichiometric interactions are formed even for low affinity interactions. Our approach should enable the structural analysis of weak interactions by NMR or X-ray crystallography to better understand the role of posttranslational modifications in dynamic biological processes.


Risky play and children's safety: balancing priorities for optimal child development.
Injury prevention plays a key role in keeping children safe, but emerging research suggests that imposing too many restrictions on children's outdoor risky play hinders their development. We explore the relationship between child development, play, and conceptions of risk taking with the aim of informing child injury prevention. Generational trends indicate children's diminishing engagement in outdoor play is influenced by parental and societal concerns. We outline the importance of play as a necessary ingredient for healthy child development and review the evidence for arguments supporting the need for outdoor risky play, including: (1) children have a natural propensity towards risky play; and, (2) keeping children safe involves letting them take and manage risks. Literature from many disciplines supports the notion that safety efforts should be balanced with opportunities for child development through outdoor risky play. New avenues for investigation and action are emerging seeking optimal strategies for keeping children "as safe as necessary," not "as safe as possible." This paradigm shift represents a potential for epistemological growth as well as cross-disciplinary collaboration to foster optimal child development while preserving children's safety.


Binge-pattern alcohol consumption and genetic risk as determinants of alcohol-related liver disease.
Alcohol-related liver disease (ARLD) represents a major public health burden. Identification of high-risk individuals would allow efficient targeting of public health interventions. Here, we show significant interactions between pattern of drinking, genetic predisposition (polygenic risk score, PRS) and diabetes mellitus, and risk of incident ARLD, in 312,599 actively drinking adults in UK Biobank. Binge and heavy binge drinking significantly increase the risk of alcohol-related cirrhosis (ARC), with higher genetic predisposition further amplifying the risk. Further, we demonstrate a pronounced interaction between heavy binge drinking and high PRS, resulting in a relative excess risk due to interaction (RERI) of 6.07. Diabetes consistently elevates ARC risk across all drinking and PRS categories, and showed significant interaction with both binge patterns and genetic risk. Overall, we demonstrate synergistic effects of binge drinking, genetics, and diabetes on ARC, with potential to identify high-risk individuals for targeted interventions.


Validation study of a new semi-automated software program for CT body composition analysis.
Computed tomography (CT) has been increasingly used to quantify abdominal muscle and fat in clinical research studies, and multiple studies have shown importance of body composition in predicting clinical outcome. The purpose of study is to compare newly developed semi-automated software (BodyCompSlicer) to commercially available validated software (Slice-O-Matic) for CT body composition analysis. CT scans of abdomen at L3 level in 30 patients were analyzed by two reviewers and using two softwares (BodyCompSlicer and Slice-O-Matic). Body composition analysis using BodyCompSlicer was semi-automated. The program automatically segmented subcutaneous fat (SF), skeletal muscle (SM), and visceral fat (VF) areas. Reviewers manually corrected the segmentation using computer-mouse interface as necessary. Body composition analysis using Slice-O-Matic was performed by manually segmenting each area using computer-mouse interface (brush tool). After segmentation, SM, SF, and VF areas were calculated using CT attenuation thresholds. Inter-observer and inter-software variability of measurements were analyzed using intraclass correlation coefficients (ICC) and coefficient of variation (COV). Inter-observer ICC and COV using BodyCompSlicer were 0.997 and 1.5% for SM, 1.000 and 0.8% for SF, and 1.000 and 1.0% for VF, whereas those using Slice-O-Matic were 0.993 and 2.5% for SM, 0.995 and 3.1% for SF, and 0.999 and 2.3% for VF. Inter-software ICCs and COV were 0.995-0.995 and 2.0-2.1% for SM, 0.991-0.994 and 3.4-3.9% for SF, and 0.998-0.998 and 2.8-3.3% for VF. Time to analyze 30 cases was 70-100 min and 150-180 min using BodyCompSlicer and Slice-O-Matic, respectively. BodyCompSlicer is comparable to Slice-O-Matic for CT body composition analysis.


Applications of gold nanoparticles in virus detection.
Viruses are the smallest known microbes, yet they cause the most significant losses in human health. Most of the time, the best-known cure for viruses is the innate immunological defense system of the host; otherwise, the initial prevention of viral infection is the only alternative. Therefore, diagnosis is the primary strategy toward the overarching goal of virus control and elimination. The introduction of a new class of nanoscale materials with multiple unique properties and functions has sparked a series of breakthrough applications. Gold nanoparticles (AuNPs) are widely reported to guide an impressive resurgence in biomedical and diagnostic applications. Here, we review the applications of AuNPs in virus testing and detection. The developed AuNP-based detection techniques are reported for various groups of clinically relevant viruses with a special focus on the applied types of bio-AuNP hybrid structures, virus detection targets, and assay modalities and formats. We pay particular attention to highlighting the functional role and activity of each core Au nanostructure and the resultant detection improvements in terms of sensitivity, detection range, and time. In addition, we provide a general summary of the contributions of AuNPs to the mainstream methods of virus detection, technical measures, and recommendations required in guidance toward commercial in-field applications.


Clades of huge phages from across Earth's ecosystems.
Bacteriophages typically have small genomes1 and depend on their bacterial hosts for replication2. Here we sequenced DNA from diverse ecosystems and found hundreds of phage genomes with lengths of more than 200 kilobases (kb), including a genome of 735 kb, which is-to our knowledge-the largest phage genome to be described to date. Thirty-five genomes were manually curated to completion (circular and no gaps). Expanded genetic repertoires include diverse and previously undescribed CRISPR-Cas systems, transfer RNAs (tRNAs), tRNA synthetases, tRNA-modification enzymes, translation-initiation and elongation factors, and ribosomal proteins. The CRISPR-Cas systems of phages have the capacity to silence host transcription factors and translational genes, potentially as part of a larger interaction network that intercepts translation to redirect biosynthesis to phage-encoded functions. In addition, some phages may repurpose bacterial CRISPR-Cas systems to eliminate competing phages. We phylogenetically define the major clades of huge phages from human and other animal microbiomes, as well as from oceans, lakes, sediments, soils and the built environment. We conclude that the large gene inventories of huge phages reflect a conserved biological strategy, and that the phages are distributed across a broad bacterial host range and across Earth's ecosystems.


Environmental Toxicant Induced Epigenetic Transgenerational Inheritance of Prostate Pathology and Stromal-Epithelial Cell Epigenome and Transcriptome Alterations: Ancestral Origins of Prostate Disease.
Prostate diseases include prostate cancer, which is the second most common male neoplasia, and benign prostatic hyperplasia (BPH), which affects approximately 50% of men. The incidence of prostate disease is increasing, and some of this increase may be attributable to ancestral exposure to environmental toxicants and epigenetic transgenerational inheritance mechanisms. The goal of the current study was to determine the effects that exposure of gestating female rats to vinclozolin has on the epigenetic transgenerational inheritance of prostate disease, and to characterize by what molecular epigenetic mechanisms this has occurred. Gestating female rats (F0 generation) were exposed to vinclozolin during E8-E14 of gestation. F1 generation offspring were bred to produce the F2 generation, which were bred to produce the transgenerational F3 generation. The transgenerational F3 generation vinclozolin lineage males at 12 months of age had an increased incidence of prostate histopathology and abnormalities compared to the control lineage. Ventral prostate epithelial and stromal cells were isolated from F3 generation 20-day old rats, prior to the onset of pathology, and used to obtain DNA and RNA for analysis. Results indicate that there were transgenerational changes in gene expression, noncoding RNA expression, and DNA methylation in both cell types. Our results suggest that ancestral exposure to vinclozolin at a critical period of gestation induces the epigenetic transgenerational inheritance of prostate stromal and epithelial cell changes in both the epigenome and transcriptome that ultimately lead to prostate disease susceptibility and may serve as a source of the increased incidence of prostate pathology observed in recent years.


The FTZ-F1 gene encodes two functionally distinct nuclear receptor isoforms in the ectoparasitic copepod salmon louse (Lepeophtheirus salmonis).
The salmon louse, Lepeophtheirus salmonis, is an ectoparasitic crustacean that annually inflicts substantial losses to the aquaculture industry in the northern hemisphere and poses a threat to the wild populations of salmonids. The salmon louse life cycle consists of eight developmental stages each separated by a molt. Fushi Tarazu Factor-1 (FTZ-F1) is an ecdysteroid-regulated gene that encodes a member of the NR5A family of nuclear receptors that is shown to play a crucial regulatory role in molting in insects and nematodes. Characterization of an FTZ-F1 orthologue in the salmon louse gave two isoforms named αFTZ-F1 and βFTZ-F1, which are identical except for the presence of a unique N-terminal domain (A/B domain). A comparison suggest conservation of the FTZ-F1 gene structure among ecdysozoans, with the exception of nematodes, to produce isoforms with unique N-terminal domains through alternative transcription start and splicing. The two isoforms of the salmon louse FTZ-F1 were expressed in different amounts in the same tissues and showed a distinct cyclical expression pattern through the molting cycle with βFTZ-F1 being the highest expressed isoform. While RNA interference knockdown of βFTZ-F1 in nauplius larvae and in pre-adult males lead to molting arrest, knockdown of βFTZ-F1 in pre-adult II female lice caused disruption of oocyte maturation at the vitellogenic stage. No apparent phenotype could be observed in αFTZ-F1 knockdown larvae, or in their development to adults, and no genes were found to be differentially expressed in the nauplii larvae following αFTZ-F1 knockdown. βFTZ-F1 knockdown in nauplii larvae caused both down and upregulation of genes associated with proteolysis and chitin binding and affected a large number of genes which are in normal salmon louse development expressed in a cyclical pattern. This is the first description of FTZ-F1 gene function in copepod crustaceans and provides a foundation to expand the understanding of the molecular mechanisms of molting in the salmon louse and other copepods.


Cryoplasty versus conventional angioplasty in femoropopliteal arterial recanalization: 3-year analysis of reintervention-free survival by treatment received.
To compare long-term efficacy of cryoplasty therapy versus conventional angioplasty in the treatment of peripheral arterial atherosclerotic stenosis on the basis of our 3-year clinical experience. From January 2006 to December 2008, a total of 155 patients with 192 lesions of the femoropopliteal sector were randomized to receive either cryoplasty or conventional balloon angioplasty. The primary study end point was lesion target patency. Follow-up with clinical evaluation of patient's symptoms, ankle-brachial index, and Doppler ultrasound was scheduled at 1, 6, 9, 12, 24, and 36 months. For the cryoplasty group (n = 86), technical immediate success was achieved in 74.4% of lesions. Rate of significant dissection was 13.5% and rate of stent placement of 22%. In the long term, target lesion patency rate at 6 months was 59.4%, with rates of 55.9, 52.6, and 49.1% at 1, 2, and 3 years, respectively. For the conventional angioplasty group (n = 69), the immediate technical success rate was 83.7%. Rate of significant dissection was 19%, and rate of stent placement was 72.9%. Patency rates at 6 months and at 1, 2, and 3 years were 71.5, 61.2, 60, and 56%, respectively. Compared with conventional angioplasty, cryoplasty showed good immediate success rates with lower stent placement rates. During the 3-year follow-up, patency rates tended to equalize between the two modalities.


Current challenges for detection of circulating tumor cells and cell-free circulating nucleic acids, and their characterization in non-small cell lung carcinoma patients. What is the best blood substrate for personalized medicine?
The practice of "liquid biopsy" as a diagnostic, prognostic and theranostic tool in non-small cell lung cancer (NSCLC) patients is an appealing approach, at least in theory, since it is noninvasive and easily repeated. In particular, this approach allows patient monitoring during treatment, as well as the detection of different genomic alterations that are potentially accessible to targeted therapy or are associated with treatment resistance. However, clinical routine practice is slow to adopt the liquid biopsy. Several reasons may explain this: (I) the vast number of methods described for potential detection of circulating biomarkers, without a consensus on the ideal technical approach; (II) the multiplicity of potential biomarkers for evaluation, in particular, circulating tumor cells (CTCs) vs. circulating tumor DNA (ctDNA); (III) the difficulty in controlling the pre-analytical phase to obtain robust and reproducible results; (IV) the present cost of the currently available techniques, which limits accessibility to patients; (V) the turnaround time required to obtain results that are incompatible with the urgent need for delivery of treatment. The purpose of this review is to describe the main advances in the field of CTC and ctDNA detection in NSCLC patients and to compare the main advantages and disadvantages of these two approaches.


DNA hypermethylation within TERT promoter upregulates TERT expression in cancer.
Replicative immortality is a hallmark of cancer cells governed by telomere maintenance. Approximately 90% of human cancers maintain their telomeres by activating telomerase, driven by the transcriptional upregulation of telomerase reverse transcriptase (TERT). Although TERT promoter mutations (TPMs) are a major cancer-associated genetic mechanism of TERT upregulation, many cancers exhibit TERT upregulation without TPMs. In this study, we describe the TERT hypermethylated oncological region (THOR), a 433-bp genomic region encompassing 52 CpG sites located immediately upstream of the TERT core promoter, as a cancer-associated epigenetic mechanism of TERT upregulation. Unmethylated THOR repressed TERT promoter activity regardless of TPM status, and hypermethylation of THOR counteracted this repressive function. THOR methylation analysis in 1,352 human tumors revealed frequent (>45%) cancer-associated DNA hypermethylation in 9 of 11 (82%) tumor types screened. Additionally, THOR hypermethylation, either independently or along with TPMs, accounted for how approximately 90% of human cancers can aberrantly activate telomerase. Thus, we propose that THOR hypermethylation is a prevalent telomerase-activating mechanism in cancer that can act independently of or in conjunction with TPMs, further supporting the utility of THOR hypermethylation as a prognostic biomarker.


[Laboratory surveillance for invasive meningococcal disease in Chile, 2006-2012].
Laboratory surveillance of Invasive Meningococcal Disease (IMD) is performed by the Institute of Public Health of Chile. It confirms identification, classifies in serogroups and analyzes the genetic profiles of Neisseria meningitidis isolates from laboratories throughout the country. To show the results of this surveillance from 2006 to 2012. A descriptive data analysis of the confirmed cases of IMD and serological characterization, susceptibility and genetic profiles of the isolates. The analysis was disaggregated by serogroup, age and region. From 2006 to 2012, 486 isolates of N. meningitidis were confirmed. In 2011 a rise in IMD rates was observed due to an increase in W serogroup cases, mainly affecting children aged 5 years or less. Serogroup W became the most prevalent during 2012 (58.3%), replacing the historically prevalent serogroup B. Predominating strains belonged to ST-32 complex/ET-5 complex (40, 4% of strains) and ST-41/44 complex/ Lineage 3 (45, 9% of strains). Laboratory surveillance has allowed the early detection of increasing IMD caused by serogroup W, which is emergent in Chile. This information has reinforced the daily monitoring of new cases, in collaboration with all the clinical laboratories of the country.


Actin polymerization-dependent activation of Cas-L promotes immunological synapse stability.
The immunological synapse formed between a T-cell and an antigen-presenting cell is important for cell-cell communication during T-cell-mediated immune responses. Immunological synapse formation begins with stimulation of the T-cell receptor (TCR). TCR microclusters are assembled and transported to the center of the immunological synapse in an actin polymerization-dependent process. However, the physical link between TCR and actin remains elusive. Here we show that lymphocyte-specific Crk-associated substrate (Cas-L), a member of a force sensing protein family, is required for transport of TCR microclusters and for establishing synapse stability. We found that Cas-L is phosphorylated at TCR microclusters in an actin polymerization-dependent fashion. Furthermore, Cas-L participates in a positive feedback loop leading to amplification of Ca2+ signaling, inside-out integrin activation, and actomyosin contraction. We propose a new role for Cas-L in T-cell activation as a mechanical transducer linking TCR microclusters to the underlying actin network and coordinating multiple actin-dependent structures in the immunological synapse. Our studies highlight the importance of mechanotransduction processes in T-cell-mediated immune responses.


Measuring social difficulties in routine patient-centred assessment: a Rasch analysis of the social difficulties inventory.
Social difficulties may add to the psychological burden experienced by cancer patients. Therefore identifying social difficulties in routine oncology practice may help prevent or alleviate distress. The Social Difficulties Inventory (SDI) is a short questionnaire developed for assessing social difficulties in cancer patients. Although well-validated, not enough is known about the clinical meaning and utility of the instrument or whether the items can be meaningfully summed to form a summary index of "Social Distress". To determine, using Rasch analysis, whether the SDI could be used as a summary index of social distress specifically examining three fundamental criteria: item fit, unidimensionality and item invariance. The Partial Credit Model was applied to a heterogeneous sample of cancer patients (n = 609) who had completed the SDI. Five items were identified as misfitting (infit mean square > or = 1.3 and standardised t-statistic > or = 2) and excluded from the subsequent analysis. The remaining items formed a unidimensional interval scale with no additional factors identified in a principal components analysis of the residuals. No differential item functioning was observed for age, gender, diagnosis, extent of disease or social deprivation. The 16-item SDI can be summed to produce an overall index of social distress, facilitating routine identification of social difficulties. Subsequent work is needed to evaluate whether the instrument is able to identify patients with high levels of social distress requiring intervention.


An Interprofessional Education Simulation Workshop: Health Professions Learning Palliative Care Communication.
Patients with life-limiting illnesses need health professionals who can communicate with each other, as well as with patients and family members. Nursing faculty teach these skills in a variety of formats and, increasingly, via simulation experiences. This pilot study aimed to compare a group of interprofessional health professions students' (N = 73) self-reported level of confidence in communication, explore behavior change and professional identity, and identify areas for future interprofessional education. Students participated in a simulated team meeting with a standardized family member of an older adult patient hospitalized with an acute aspiration pneumonia and a chronic, progressive illness. Postworkshop, students rated themselves as significantly more confident in interprofessional and palliative care communication (p ⩽ .001) than preworkshop, identified important areas of behavior change and professional identity, and provided faculty with ideas for future simulation workshops. Additional research is needed regarding longitudinal curricular efforts and direct patient care outcomes. [J Nurs Educ. 2018;57(8):493-497.].