PAX1 Methylation as a Potential Biomarker to Predict the Progression of Cervical Intraepithelial Neoplasia: A Meta-analysis of Related Studies.
The methylation of paired box gene 1 (PAX1) has a great influence on the process of cervical lesion. However, available evidence for the association between PAX1 methylation and cervical intraepithelial neoplasia (CIN) are inconsistent. Here, we systematically reviewed and analyzed PAX1 methylation in progress of CIN. Two investigators independently searched eligible studies of PAX1 methylation and CIN that were published in PubMed, Cochrane Library, EMBASE, and Web of Science databases until November 30, 2016. We extracted clinicopathologic features of CIN and cervical cancel relevant to PAX1 methylation. Odds ratios (ORs) with their 95% confidence intervals (CIs) were used to assess the association between PAX1 methylation and progression of patients with CIN. Seven studies composed of 1055 patients with various stages of CIN and cervical cancel were eventually included. The results revealed that PAX1 methylation was associated with transition of CIN I to CIN II/III (OR, 0.09; 95% CI, 0.04-0.19) and CIN II/III to cervical cancer (OR, 0.16; 95% CI, 0.05-0.46), and similar results were produced in sensitivity analysis. Also, we found that the OR value was associated with average age and number of patients, publication year, and study location of included articles. PAX1 gene methylation was associated with the transition of CIN I to CIN II/III and CIN II/III to cervical cancer, so that it could be an auxiliary biomarker to estimate the risk of CIN progress. Moreover, PAX1 may help to determine appropriate reexaminations and treatment for patients with various stages of CIN.


Psychosocial interventions in stimulant use disorders: a systematic review and qualitative synthesis of randomized controlled trials.
Stimulant use disorders are highly prevalent with a large burden of disease. Most clinical guidelines recommend psychosocial interventions, but there are no clear hierarchies or indications. Moreover, these interventions have been reported unevenly in the literature. Identifying the most suitable treatment for each patient therefore represents a major challenge. In this review, we describe all psychosocial interventions for stimulant use disorders investigated in randomized controlled trials - including contingency management, cognitive behavioral interventions, community reinforcement approach, 12-step program, meditation-based interventions and physical exercise, supportive expressive psychodynamic therapy, interpersonal psychotherapy, family therapy, motivational interviewing, drug counseling -, and we synthesize the main findings of these studies. Similarities and differences between treatments are highlighted, suggesting that distinct psychosocial interventions can be relevant for certain patients' groups but not for others. Conversely, several interventions can be equally effective in similar clinical contexts, suggesting that a shared element such as therapeutic alliance is key. Finally, combined approaches emerge as a viable option for people with complex needs. Future studies will need to benchmark psychosocial interventions in stimulant use disorders and ascertain markers of response with a view to individualized treatment.


Early phonological and sociocognitive skills as predictors of later language and social communication outcomes.
Previous studies of outcome for children with early language delay have focused on measures of early language as predictors of language outcome. This study investigates whether very early processing skills (VEPS) known to underpin language development will be better predictors of specific language and social communication outcomes than measures of language itself. Participants were 163 children referred to clinical services with concerns about language at 2;6-3;6 years and followed up at 4-5 years. Novel assessments of phonological and sociocognitive processing were administered at Time 1 (T1), together with a standardised test of receptive and expressive language, and parental report of expressive vocabulary. The language test was re-administered at Time 2 (T2), together with assessments of morphosyntax and parental reports of social communication. Intercorrelations at and between T1 and T2 were high, and dissociations were rare. Ordinal regressions were run, entering predictors singly and simultaneously. With the exception of the phonological task, every early measure on its own was significantly predictive of most outcomes, and receptive language was the strongest all-round predictor. Results of simultaneous entry, controlling for the effect of other predictors, showed that early language was the strongest predictor of general language outcome, but early phonology was the strongest predictor of a measure of morphosyntax, and early sociocognition the strongest predictor of social communication. Language measures which draw on a wide range of skills were the strongest overall predictors of general language outcomes. However, our VEPS measures were stronger predictors of specific outcomes. The clinical and theoretical implications of these findings are discussed.


Choreography of early thalamocortical development.
Thalamic axons, which carry most of the information from the sensory environment, are amongst the first projections to reach the cerebral cortex during embryonic development. It has been proposed that the scaffold of early generated cells in the ventral thalamus, internal capsule and preplate play a pivotal role in their deployment through sharp gene expression boundaries. These ideas were recently evaluated in various strains of mutant mice. In Tbr1, Gbx2, Pax6 KO both thalamic and corticofugal projections fail to traverse the striatocortical junction. In both Emx2 and Pax6 KO brains, the misrouted thalamic afferents are accompanied by displacements of the pioneering projections from the internal capsule. Regardless of their altered route, thalamic afferents in the reeler and L1 KO mice seem to be able to redistribute themselves on the cortical sheet and establish normal periphery-related representation in the somatosensory cortex. Early neural activity delivered through the thalamic projections is thought to be involved in the realignment process of thalamic axons at the time of their accumulation in the subplate layer. However, axonal growth and the early topographic arrangement of thalamocortical fiber pathways appear normal in the Snap25 KO, where action potential mediated synaptic vesicle release is disrupted. We therefore suggest that intercellular communication mediated by constitutive secretion of transmitters or growth factors might play a dominant role during early thalamocortical development.


Rapid release of tissue enzymes into blood after blast exposure: potential use as biological dosimeters.
Explosive blast results in multiple organ injury and polytrauma, the intensity of which varies with the nature of the exposure, orientation, environment and individual resilience. Blast overpressure alone may not precisely indicate the level of body or brain injury after blast exposure. Assessment of the extent of body injury after blast exposure is important, since polytrauma and systemic factors significantly contribute to blast-induced traumatic brain injury. We evaluated the activity of plasma enzymes including aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and creatine kinase (CK) at different time points after blast exposure using a mouse model of single and repeated blast exposures to assess the severity of injury. Our data show that activities of all the enzymes in the plasma were significantly increased as early as 1 h after blast exposure. The elevated enzyme activity remained up to 6 h in an overpressure dose-dependent manner and returned close to normal levels at 24 h. Head-only blast exposure with body protection showed no increase in the enzyme activities suggesting that brain injury alone does not contribute to the systemic increase. In contrast to plasma increase, AST, ALT and LDH activity in the liver and CK in the skeletal muscle showed drastic decrease at 6 h after blast exposures. Histopathology showed mild necrosis at 6 h and severe necrosis at 24 h after blast exposures in liver and no changes in the skeletal muscle suggesting that the enzyme release from the tissue to plasma is probably triggered by transient cell membrane disruption from shockwave and not due to necrosis. Overpressure dependent transient release of tissue enzymes and elevation in the plasma after blast exposure suggest that elevated enzyme activities in the blood can be potentially used as a biological dosimeter to assess the severity of blast injury.


Metabolism of orally administered rac-1-O-[1'-14C]dodecylglycerol and nutritional effects of dietary rac-1-O-dodecylglycerol in mice.
The metabolism of orally administered rac-1-O-[1'-14C]dodecylglycerol was investigated in mice. The substrate was rapidly absorbed in the intestine and then incorporated into ether glycerolipids of various organs and tissues in high proportions. In intestine and liver, however, large amounts of rac-1-O-[1'-14C]dodecylglycerol were catabolized by oxidative cleavage of the ether bond followed by degradation of the radioactive cleavage product, i.e., lauric acid, to water-soluble metabolites that were excreted in the urine at a fast rate. The feeding of a rac-1-O-dodecylglycerol-containing diet (1 g rac-1-O-dodecylglycerol/kg body weight X day) given over a period of 4 weeks did not significantly alter body weights or organ weights of mice. Analysis of total lipids revealed that high proportions of the substrate were incorporated into ether lipids of all organs and tissues during the feeding period, generally accompanied by a remarkable increase in saturated acyl moieties and a concomitant decrease of linoleoyl moieties of total lipids. Yet, 4 weeks after removal of the rac-1-O-dodecylglycerol-containing diet, the lipids of murine organs and tissues showed a close resemblance to those of the control group.


Culture and carelessness: constituting disability in South India.
Professional and lay explanations of disability, collected via interviews and participant-observation during fieldwork in Hyderabad, South India, identify "carelessness" and "superstition" as major impediments to good health among the general population, and education as the key solution. In that such findings suggest a valorization of personal responsibility for self-care, the Foucauldian concept of biopower appeared a salient framework for analysis. Although illuminating, however, biopower was ultimately inadequate for explaining what emerged, on closer analysis, as significant discrepancies between assumptions about how disabled people engaged with healthcare services and their actual beliefs and practices; and between the moral interpretations different stakeholders made of "carelessness" in describing perceived causes of disability. My data also suggested that education was not in itself a key determinant in people's healthcare decisions. This article explores these differences between official and demotic discourses concerning the causes of disability and attempts to account for them ethnographically.


Safety and efficacy of video-assisted versus conventional lung resection for lung cancer.
We sought to evaluate the use of video-assisted thoracoscopy among patients with lung cancer and its safety and effectiveness relative to conventional resection. A cohort study (1994-2002) was conducted by using the Surveillance, Epidemiology, and End-Results Medicare database. Video-assisted thoracoscopy and conventional resection were hypothesized to be equivalent in terms of risks of death. Equivalency was defined by a confidence interval of 0.72 to 1.28 for the odds of 30-day death and 0.89 to 1.11 for the hazard of death, corresponding to a difference of no more than 1% for 30-day mortality and 5% for 5-year survival, respectively. Among 12,958 patients who underwent segmentectomy or lobectomy (mean age, 74 +/- 5 years), 6% underwent video-assisted thoracoscopy. The use of video-assisted thoracoscopy increased from 1% to 9% between 1994 and 2002. Compared with those who underwent conventional resection, patients who underwent video-assisted thoracoscopy more frequently had smaller tumors (P < .001) and stage I disease (P = .03), underwent lymphadenectomy (P < .001), and were cared for by higher-volume surgeons (P < .001) and at higher-volume hospitals (P < .001). After adjusting for differences in patient, cancer, management, and provider characteristics, the odds of early death were not significantly different between patients undergoing video-assisted thoracoscopy and those undergoing conventional resection, although equivalency was not demonstrated (adjusted odds ratio, 0.93; 95% confidence interval, 0.57-1.50). The hazard of death was equivalent for video-assisted thoracoscopy and conventional resection (adjusted hazard ratio, 0.99; 95% confidence interval, 0.90-1.08). Video-assisted thoracoscopy was uncommonly used to manage lung cancer, although its use has increased over time. Video-assisted thoracoscopy and conventional resection were equivalent in terms of long-term survival.


Neuropeptides, Inflammation, Biofilms, and diabetic Foot Ulcers.
A diabetic foot ulcer (DFU) is a serious complication in patients with diabetes mellitus (DM). A DFU is the most common cause of non-traumatic limb amputation, and patients with DFUs have increased mortality rates within 5 years after amputation. DFUs also increase the risk of cardiovascular and cerebrovascular diseases; therefore, with the increasing incidence and prevalence of diabetic foot wounds, DFUs are gradually becoming a major public health problem. The pathophysiology of DFUs is complicated and remains unclear. In recent years, many studies have demonstrated that the pathophysiology of DFUs is especially associated with neuropeptides, inflammation, and biofilms. Neuropeptides, especially substance P (SP) and calcitonin gene-related peptide (CGRP), play an important role in wound healing. SP and CGRP accelerate the healing of cutaneous wounds by promoting neovascularization, inhibiting the release of certain proinflammatory chemokines, regulating macrophage polarization, and so on. However, the expression of SP and CGRP was downregulated in DM and DFUs. DFUs are characterized by a sustained inflammatory phase. Immune cells such as neutrophils and macrophages are involved in the sustained inflammatory phase in DFUs by extracellular traps (NETs) and dysregulated macrophage polarization, which delays wound healing. Furthermore, DFUs are at increased risk of biofilm formation. Biofilms disturb wound healing by inducing a chronic inflammatory response, inhibiting macrophage phagocytosis and keratinocyte proliferation migration, and transferring antimicrobial resistance genes. To understand the relationships among neuropeptides, inflammation, biofilms, and DFUs, this review highlights the recent scientific advances that provide possible pathophysiological insights into the delayed healing of DFUs.


Moderation analysis exploring associations between age and mucocutaneous activity in Behçet's syndrome: A multicenter study from Turkey.
The aim of the present study was to examine the effects of age on mucocutaneous activity by using moderation analysis in Behçet's syndrome (BS). In this cross-sectional study, 887 BS patients (female : male, 481:406; mean age, 38.4 ± 10.9 years) followed in 13 tertiary centers in Turkey were included. Mucocutaneous activity was evaluated by using the Mucocutaneous Index (MI) according to sex and disease course. Moderation analysis was performed to test the effect of age on mucocutaneous activity. A moderator variable is a third variable and affects the relationship between independent and outcome variables. Age was chosen as a potential moderator variable (interaction effect), MI score as the outcome variable and sex as an independent variable in the analysis. The moderation analysis tested the effects of age in three steps: whole BS patient group, patients without systemic involvement and those with systemic involvement. The moderation model was only significant in BS patients with systemic involvement (P = 0.0351), and a significant relationship was observed between female sex and MI score (P = 0.0156). In addition, the interaction plot showed that female patients had increased MI scores compared with male patients, especially in the 28-year-old age group (P = 0.0067). Moreover, major organ involvement was newly diagnosed in the majority of these young female BS patients. Our results suggest that the relationship between sex and mucocutaneous activity was moderated by age in the systemic involvement group. Also, increased mucocutaneous activity may be associated with new major organ involvement in young female BS patients with systemic involvement.


Discovery of a Potent and Selective CDKL5/GSK3 Chemical Probe That Is Neuroprotective.
Despite mediating several essential processes in the brain, including during development, cyclin-dependent kinase-like 5 (CDKL5) remains a poorly characterized human protein kinase. Accordingly, its substrates, functions, and regulatory mechanisms have not been fully described. We realized that availability of a potent and selective small molecule probe targeting CDKL5 could enable illumination of its roles in normal development as well as in diseases where it has become aberrant due to mutation. We prepared analogs of AT-7519, a compound that has advanced to phase II clinical trials and is a known inhibitor of several cyclin-dependent kinases (CDKs) and cyclin-dependent kinase-like kinases (CDKLs). We identified analog 2 as a highly potent and cell-active chemical probe for CDKL5/GSK3 (glycogen synthase kinase 3). Evaluation of its kinome-wide selectivity confirmed that analog 2 demonstrates excellent selectivity and only retains GSK3α/β affinity. We next demonstrated the inhibition of downstream CDKL5 and GSK3α/β signaling and solved a co-crystal structure of analog 2 bound to human CDKL5. A structurally similar analog (4) proved to lack CDKL5 affinity and maintain potent and selective inhibition of GSK3α/β, making it a suitable negative control. Finally, we used our chemical probe pair (2 and 4) to demonstrate that inhibition of CDKL5 and/or GSK3α/β promotes the survival of human motor neurons exposed to endoplasmic reticulum stress. We have demonstrated a neuroprotective phenotype elicited by our chemical probe pair and exemplified the utility of our compounds to characterize the role of CDKL5/GSK3 in neurons and beyond.


Association between HACEK bacteraemia and endocarditis.
We retrospectively examined medical records of 87 patients with bacteraemia caused by members of the HACEK group (Haemophilus parainfluenzae, Aggregatibacter actinomycetemcomitans, Aggregatibacter aphrophilus, Aggregatibacter paraphrophilus, Cardiobacterium spp., Eikenella corrodens and Kingella spp.) to determine whether endocarditis was present, as defined by the Duke criteria. The overall positive predictive value (PPV) of HACEK bacteraemia for endocarditis was 60 %. The PPV varied with different HACEK species from 0 % (E. corrodens) to 100 % (A. actinomycetemcomitans).


Variation in thermal performance and reaction norms among populations of Drosophila melanogaster.
The major goal of evolutionary thermal biology is to understand how variation in temperature shapes phenotypic evolution. Comparing thermal reaction norms among populations from different thermal environments allows us to gain insights into the evolutionary mechanisms underlying thermal adaptation. Here, we have examined thermal adaptation in six wild populations of the fruit fly (Drosophila melanogaster) from markedly different natural environments by analyzing thermal reaction norms for fecundity, thorax length, wing area, and ovariole number under ecologically realistic fluctuating temperature regimes in the laboratory. Contrary to expectation, we found only minor differences in the thermal optima for fecundity among populations. Differentiation among populations was mainly due to differences in absolute (and partly also relative) thermal fecundity performance. Despite significant variation among populations in the absolute values of morphological traits, we observed only minor differentiation in their reaction norms. Overall, the thermal reaction norms for all traits examined were remarkably similar among different populations. Our results therefore suggest that thermal adaptation in D. melanogaster predominantly involves evolutionary changes in absolute trait values rather than in aspects of thermal reaction norms.


Adaptive evolution of pelvic reduction in sticklebacks by recurrent deletion of a Pitx1 enhancer.
The molecular mechanisms underlying major phenotypic changes that have evolved repeatedly in nature are generally unknown. Pelvic loss in different natural populations of threespine stickleback fish has occurred through regulatory mutations deleting a tissue-specific enhancer of the Pituitary homeobox transcription factor 1 (Pitx1) gene. The high prevalence of deletion mutations at Pitx1 may be influenced by inherent structural features of the locus. Although Pitx1 null mutations are lethal in laboratory animals, Pitx1 regulatory mutations show molecular signatures of positive selection in pelvic-reduced populations. These studies illustrate how major expression and morphological changes can arise from single mutational leaps in natural populations, producing new adaptive alleles via recurrent regulatory alterations in a key developmental control gene.


Imbalances in the health labour force: an assessment using data from three national health facility surveys.
Accurate knowledge of the characteristics of the health labour force that can affect health care production is of critical importance to health planners and policymakers. This study uses health facility survey data to examine characteristics of the primary health care labour force in Nicaragua, Tanzania and Bangladesh. The characteristics examined are those that are likely to affect service provision, including urban/rural distribution, demographic characteristics, and experience and in-service training, for three types of providers (physicians, nurses and auxiliary nurses). The profiles suggest a pattern of urban/rural imbalances in Nicaragua and Tanzania. The Bangladesh facility survey did not include hospitals, thereby making concrete conclusions on the supply and distribution of providers difficult to make. Multivariate logistic regressions are used to assess the relationship between the urban/rural placement of providers by health need, population demand and facility characteristics. Health need, as measured by child mortality rates, does not have a significant association with the placement of providers in either country, unlike population size and annual growth rates. The mean number of years providers have worked at a facility is significantly associated with a decreased likelihood of rural placement for the three types of providers in Nicaragua. The mean age and percentage of female providers at each facility has a negative association with the placement of rural providers in Tanzania. The use of health facility data to profile the health care labour force is also discussed.


Treatment of infantile Pompe disease with alglucosidase alpha: the UK experience.
Treatment of infantile Pompe disease with recombinant human acid α-glucosidase has shown substantial improvement in survival, and in cardiac, motor and respiratory functions. We analyzed the outcome of all patients with infantile Pompe disease treated in the United Kingdom since the availability of the enzyme, using a questionnaire-based survey circulated to all treating centres. A total of 20 infants were treated from 2000 to 2009. Median ages at diagnosis and treatment were 5.75 months (range 0.25-31 months) and 6.5 months (0.5-32 months), respectively. Median duration of treatment was 31 months (1-102 months). Overall ventilator-free survival was 35% (7/20 infants), while 35% (7/20) died at a median age of 10 months (5.75-15 months) and 30% (6/20) were alive but ventilator-dependent. Endotracheal intubation for acute deterioration carried a high risk of failure of extubation and progression to long-term ventilation (LTV), but elective general anaesthesia, in contrast, was well tolerated. Overall outcome was worse than in the pivotal clinical trials; possible causes include later diagnosis and treatment in our patients and a higher incidence of infants at the severe end of the clinical spectrum. Careful consideration must be given to all possible outcomes, including LTV, before commencing enzyme replacement therapy in newly diagnosed infants.


Isolation and identification of iron-chelating peptides from casein hydrolysates.
Iron deficiency is a common nutritional disorder worldwide. Peptides derived from protein hydrolysates have recently attracted interest as novel iron chelators due to their superiority in terms of increasing solubility, bioavailability, absorption and stability. The aim of this study was to isolate and identify iron-chelating peptides from casein hydrolysates. Casein was hydrolyzed (trypsin, 3 h) and subsequently isolated using ultrafiltration and RP-HPLC. Four iron-chelating casein hydrolysate peptides, named CHP-1, CHP-2, CHP-3 and CHP-4, were identified by LC-MS/MS, and their amino acid sequences were Glu-Asp-Val-Pro-Ser-Glu-Arg (EDVPSER), His-Lys-Glu-Met-Pro-Phe-Pro-Lys (HKEMPFPK), Asn-Met-Ala-Ile-Asn-Pro-Ser-Lys (NMAINPSK) and Ala-Val-Pro-Tyr-Pro-Gln-Arg (AVPYPQR), with molecular weights of 830.6120 Da, 1012.5280 Da, 873.4440 Da and 829.4570 Da, respectively. The artificially synthesized peptides of CHP-1, CHP-2, CHP-3 and CHP-4 were verified, and their iron-chelating rates were 11.14%, 8.02%, 7.57% and 59.76%, respectively. These results suggested that the isolated iron-chelating peptides might serve as potential iron supplements and be used as food additives and functional foods.


Optogenetic stimulation of VIPergic SCN neurons induces photoperiodic-like changes in the mammalian circadian clock.
Circadian clocks play key roles in how organisms respond to and even anticipate seasonal change in day length, or photoperiod. In mammals, photoperiod is encoded by the central circadian pacemaker in the brain, the suprachiasmatic nucleus (SCN). The subpopulation of SCN neurons that secrete the neuropeptide VIP mediates the transmission of light information within the SCN neural network, suggesting a role for these neurons in circadian plasticity in response to light information that has yet to be directly tested. Here, we used in vivo optogenetic stimulation of VIPergic SCN neurons followed by ex vivo PERIOD 2::LUCIFERASE (PER2::LUC) bioluminescent imaging to test whether activation of this SCN neuron subpopulation can induce SCN network changes that are hallmarks of photoperiodic encoding. We found that optogenetic stimulation designed to mimic a long photoperiod indeed altered subsequent SCN entrained phase, increased the phase dispersal of PER2 rhythms within the SCN network, and shortened SCN free-running period-similar to the effects of a true extension of photoperiod. Optogenetic stimulation also induced analogous changes on related aspects of locomotor behaviour in vivo. Thus, selective activation of VIPergic SCN neurons induces photoperiodic network plasticity in the SCN that underpins photoperiodic entrainment of behaviour.


The E3-ubiquitin ligase TRIM50 interacts with HDAC6 and p62, and promotes the sequestration and clearance of ubiquitinated proteins into the aggresome.
In this study we report that, in response to proteasome inhibition, the E3-Ubiquitin ligase TRIM50 localizes to and promotes the recruitment and aggregation of polyubiquitinated proteins to the aggresome. Using Hdac6-deficient mouse embryo fibroblasts (MEF) we show that this localization is mediated by the histone deacetylase 6, HDAC6. Whereas Trim50-deficient MEFs allow pinpointing that the TRIM50 ubiquitin-ligase regulates the clearance of polyubiquitinated proteins localized to the aggresome. Finally we demonstrate that TRIM50 colocalizes, interacts with and increases the level of p62, a multifunctional adaptor protein implicated in various cellular processes including the autophagy clearance of polyubiquitinated protein aggregates. We speculate that when the proteasome activity is impaired, TRIM50 fails to drive its substrates to the proteasome-mediated degradation, and promotes their storage in the aggresome for successive clearance.


The statistics of identifying differentially expressed genes in Expresso and TM4: a comparison.
Analysis of DNA microarray data takes as input spot intensity measurements from scanner software and returns differential expression of genes between two conditions, together with a statistical significance assessment. This process typically consists of two steps: data normalization and identification of differentially expressed genes through statistical analysis. The Expresso microarray experiment management system implements these steps with a two-stage, log-linear ANOVA mixed model technique, tailored to individual experimental designs. The complement of tools in TM4, on the other hand, is based on a number of preset design choices that limit its flexibility. In the TM4 microarray analysis suite, normalization, filter, and analysis methods form an analysis pipeline. TM4 computes integrated intensity values (IIV) from the average intensities and spot pixel counts returned by the scanner software as input to its normalization steps. By contrast, Expresso can use either IIV data or median intensity values (MIV). Here, we compare Expresso and TM4 analysis of two experiments and assess the results against qRT-PCR data. The Expresso analysis using MIV data consistently identifies more genes as differentially expressed, when compared to Expresso analysis with IIV data. The typical TM4 normalization and filtering pipeline corrects systematic intensity-specific bias on a per microarray basis. Subsequent statistical analysis with Expresso or a TM4 t-test can effectively identify differentially expressed genes. The best agreement with qRT-PCR data is obtained through the use of Expresso analysis and MIV data. The results of this research are of practical value to biologists who analyze microarray data sets. The TM4 normalization and filtering pipeline corrects microarray-specific systematic bias and complements the normalization stage in Expresso analysis. The results of Expresso using MIV data have the best agreement with qRT-PCR results. In one experiment, MIV is a better choice than IIV as input to data normalization and statistical analysis methods, as it yields as greater number of statistically significant differentially expressed genes; TM4 does not support the choice of MIV input data. Overall, the more flexible and extensive statistical models of Expresso achieve more accurate analytical results, when judged by the yardstick of qRT-PCR data, in the context of an experimental design of modest complexity.


Alternative splicing of caspase 9 is modulated by the phosphoinositide 3-kinase/Akt pathway via phosphorylation of SRp30a.
Increasing evidence points to the functional importance of alternative splice variations in cancer pathophysiology. Two splice variants are derived from the CASP9 gene via the inclusion (Casp9a) or exclusion (Casp9b) of a four-exon cassette. Here we show that alternative splicing of Casp9 is dysregulated in non-small cell lung cancers (NSCLC) regardless of their pathologic classification. Based on these findings we hypothesized that survival pathways activated by oncogenic mutation regulated this mechanism. In contrast to K-RasV12 expression, epidermal growth factor receptor (EGFR) overexpression or mutation dramatically lowered the Casp9a/9b splice isoform ratio. Moreover, Casp9b downregulation blocked the ability of EGFR mutations to induce anchorage-independent growth. Furthermore, Casp9b expression blocked inhibition of clonogenic colony formation by erlotinib. Interrogation of oncogenic signaling pathways showed that inhibition of phosphoinositide 3-kinase or Akt dramatically increased the Casp9a/9b ratio in NSCLC cells. Finally, Akt was found to mediate exclusion of the exon 3,4,5,6 cassette of Casp9 via the phosphorylation state of the RNA splicing factor SRp30a via serines 199, 201, 227, and 234. Taken together, our findings show that oncogenic factors activating the phosphoinositide 3-kinase/Akt pathway can regulate alternative splicing of Casp9 via a coordinated mechanism involving the phosphorylation of SRp30a.


Spiral-medial butterfly fractures (AO-12-B1) in distal diaphysis of humerus with rotational forces: preliminary results of open reduction and plate-screw fixation.
Humerus shaft fractures are frequent injuries in orthopedic practice and generally occurs with direct and indirect traumas. While the most of these fractures can be managed with conservative methods, in some situations surgery is needed. The aim of this study is to define a different mechanism and treatment result of humerus spiral fractures with medial butterfly fragment (AO-12-B1). Between 2010 and 2013, 11 patients who had undergone surgery for spiral humerus shaft fracture with medial large butterfly fragment (AO type 12-B1) resulting from a motor vehicle accident were enrolled to the study. All data was retrieved from the hospital's database retrospectively. All fractures were closed type and all of the patients were treated with open reduction and plate osteosynthesis through lateral approach. Elbow and shoulder ROMs were examined for functional outcomes. The DASH (Disabilities of the Arm, Shoulder and Hand) scoring system was applied at the final follow-up visit. There were 7 male and 4 female patients with a mean age of 36 years (range, 28-50 years). All of the fractures occurred with same mechanism resulting from motor vehicle accident. Conservative treatment was not considered for any of the cases. The mean time to surgery was 2 days (1-3 days). The mean postoperative follow-up period was 14 months (6-24 months). All fractures had healed completely at 3 months postoperatively. At the final follow-up visit the mean DASH score was 4.2 (3.3-6.7). Humerus spiral shaft fractures with a large medial butterfly fragment can be seen in dashboard injuries. It is important to fix the medial large fragment anatomically and minimal invasively in order to achieve union and not to disturb the vascular supply.


Fundamental and biotechnological applications of neutron scattering measurements for macromolecular dynamics.
To explore macromolecular dynamics on the picosecond timescale, we used neutron spectroscopy. First, molecular dynamics were analyzed for the hyperthermophile malate dehydrogenase from Methanococcus jannaschii and a mesophilic homologue, the lactate dehydrogenase from Oryctolagus cunniculus muscle. Hyperthermophiles have elaborate molecular mechanisms of adaptation to extremely high temperature. Using a novel elastic neutron scattering approach that provides independent measurements of the global flexibility and of the structural resilience (rigidity), we have demonstrated that macromolecular dynamics represents one of these molecular mechanisms of thermoadaptation. The flexibilities were found to be similar for both enzymes at their optimal activity temperature and the resilience is higher for the hyperthermophilic protein. Secondly, macromolecular motions were examined in a native and immobilized dihydrofolate reductase (DHFR) from Escherichia coli. The immobilized mesophilic enzyme has increased stability and decreased activity, so that its properties are changed to resemble those of the thermophilic enzyme. Are these changes reflected in dynamical behavior? For this study, we performed quasielastic neutron scattering measurements to probe the protein motions. The residence time is 7.95 ps for the native DHFR and 20.36 ps for the immobilized DHFR. The average height of the potential barrier to local motions is therefore increased in the immobilized DHFR, with a difference in activation energy equal to 0.54 kcal/mol, which is, using the theoretical rate equation, of the same order than expected from calculation.


Adapting to a new reality: COVID-19 coronavirus and online education in the health professions.
The current novel coronavirus, COVID-19, has effected a significant change in the way industry-based and tertiary health professions education (HPE) can occur. Advice for strict, widespread social distancing has catalysed the transformation of course delivery into fully online design across nations. This is problematic for HPE, which has traditionally relied on face-to-face learner interaction, in the form of skills laboratories, simulation training and industry-based clinical placements. The transition to online-only course delivery has brought with it a need to address particular issues regarding the construction and delivery of quality curricula and education activities. It is in this context that regional, rural and remote health professionals and academics can provide invaluable insights into the use of technology to overcome the tyranny of distance, promote high-quality online HPE and enable the ongoing development of communities of practice. This article is the first in a series addressing the risks and opportunities in the current transition to online HPE, providing practical solutions for educators who are now unable to embrace more traditional face-to-face HPE delivery methods and activities.


Gender difference and effect of pharmacotherapy: findings from a smoking cessation service.
Smoking cessation services are available in England to provide assistance to those wishing to quit smoking. Data from one such service were analysed in order to investigate differences in quit rate between males and females prescribed with different treatments. A logistic regression model was fitted to the data using the binary response of self-reported quit (failed attempt = 0, successful attempt = 1), validated by Carbon Monoxide (CO) monitoring, 4 weeks after commencing programme. Main effects fitted were: client gender; age; region; the type of advisory sessions; and pharmacotherapy, Nicotine Replacement Therapy (NRT) or Varenicline. A second model was fitted including all main effects plus two-way interactions except region. These models were repeated using 12-week self-reported quit as the outcome. At 4 weeks, all main effects were statistically significant, with males more likely (odds ratio and 95 % CI, females v males = 0.88 [0.79-0.97]), older smokers more likely (adjusted odds ratios [OR] and 95 % confidence interval [CI] respectively for groups 20-29, 30-49, 50-69 and 70+ vs 12-19 age group: 1.79 [1.39-2.31], 2.12 [1.68-2.68], 2.30 [1.80-2.92] and 2.47 [1.81-3.37] and for overall difference between groups, χ2(4) = 53.5, p < 0.001) and clients being treated with Varenicline more likely to have successfully quit than those on NRT (adjusted OR and 95 % CI for Varenicline vs NRT = 1.41 [1.21-1.64]). Statistically significant interactions were observed between (i) gender and type of counselling, and (ii) age and type of counselling. Similar results were seen in relation to main effects at 12 weeks except that type of counselling was non-significant. The only significant interaction at this stage was between gender and pharmacotherapy (adjusted OR and 95 % CI for females using Varenicline versus all other groups = 1.43 [1.06-1.94]). Gender and treatment options were identified as predictors of abstinence at both 4 and 12 weeks after quitting smoking. Furthermore, interactions were observed between gender and (i) type of counselling received (ii) pharmacotherapy. In particular, the quit rate in women at 12 weeks was significantly improved in conjunction with Varenicline use. These findings have implications for service delivery.


Nanotopography regulates motor neuron differentiation of human pluripotent stem cells.
The regulation of human pluripotent stem cell (hPSC) behaviors has been mainly studied through exploration of biochemical factors. However, the current directed differentiation protocols for hPSCs that completely rely on biochemical factors remain suboptimal. It has recently become evident that coexisting biophysical signals in the stem cell microenvironment, including nanotopographic cues, can provide potent regulatory signals to mediate adult stem cell behaviors, including self-renewal and differentiation. Herein, we utilized a recently developed, large-scale nanofabrication technique based on reactive-ion etching (RIE) to generate random nanoscale structures on glass surfaces with high precision and reproducibility. We report here that hPSCs are sensitive to nanotopographic cues and such nanotopographic sensitivity can be leveraged for improving directed neuronal differentiation of hPSCs. We demonstrate early neuroepithelial conversion and motor neuron (MN) progenitor differentiation of hPSCs can be promoted using nanoengineered topographic substrates. We further explore how hPSCs sense the substrate nanotopography and relay this biophysical signal through a regulatory signaling network involving cell adhesion, the actomyosin cytoskeleton, and Hippo/YAP signaling to mediate the neuroepithelial induction of hPSCs. Our study provides an efficient method for large-scale production of MNs from hPSCs, useful for regenerative medicine and cell-based therapies.


Normalization enables robust validation of disparity estimates from neural populations.
Binocular fusion takes place over a limited region smaller than one degree of visual angle (Panum's fusional area), which is on the order of the range of preferred disparities measured in populations of disparity-tuned neurons in the visual cortex. However, the actual range of binocular disparities encountered in natural scenes extends over tens of degrees. This discrepancy suggests that there must be a mechanism for detecting whether the stimulus disparity is inside or outside the range of the preferred disparities in the population. Here, we compare the efficacy of several features derived from the population responses of phase-tuned disparity energy neurons in differentiating between in-range and out-of-range disparities. Interestingly, some features that might be appealing at first glance, such as the average activation across the population and the difference between the peak and average responses, actually perform poorly. On the other hand, normalizing the difference between the peak and average responses results in a reliable indicator. Using a probabilistic model of the population responses, we improve classification accuracy by combining multiple features. A decision rule that combines the normalized peak to average difference and the peak location significantly improves performance over decision rules based on either measure in isolation. In addition, classifiers using normalized difference are also robust to mismatch between the image statistics assumed by the model and the actual image statistics.


The Na+/K+/2Cl- cotransporter in the sea bass Dicentrarchus labrax during ontogeny: involvement in osmoregulation.
This study combines a cellular and molecular analysis of the Na(+)/K(+)/2Cl(-) cotransporter (NKCC) to determine the osmoregulatory role of this protein in different tissues during the ontogeny of the sea bass. We have characterized the complete sequence of the NKCC1 isoform isolated from the sea bass gills and have identified, by immunofluorescence, NKCC1, and other isoforms, within the epithelium of the major osmoregulatory organs. Different (absorptive and secretory) functions have been attributed to this protein according to the tissue and salinity. The effects of short- (1-4 days), medium- (7-21 days) and long (6 months)-term freshwater (FW) adaptations were investigated, in comparison with seawater (SW)-maintained sea bass. In adult sea bass after long-term adaptation to FW and SW, the gills had the highest expression of NKCC mRNA compared with the median/posterior kidney and to the posterior intestine. Expression of NKCC mRNA in the kidney was 95% (SW) and 63% (FW) lower, and in the intestine 98% (SW) and 77% (FW) lower. Compared to SW-maintained sea bass, long-term FW adaptation induced a significant 5.6-fold decrease in the branchial NKCC gene expression whereas the intestinal and renal expressions did not vary significantly. The cells of the intestine and collecting ducts as well as a part of the epithelium lining the urinary bladder expressed NKCC apically. Within the gill chloride cells, NKCC was found basolaterally in SW-acclimated fish; some apically stained cells were detected after 7 days of FW exposure and their relative number increased progressively following FW acclimation. The appearance of FW-type chloride cells induces a functional shift of the gills from a secretory to an absorptive epithelium, which was only completed after long-term exposure to FW. Short- and medium-term exposure to FW induced a progressive decrease in total NKCC content and an increase in functionally different branchial chloride cells. During development, the cotransporter was already expressed in tegumentary ionocytes and along the digestive tract of late embryos. NKCC was recorded in the branchial chamber and along the renal collecting ducts in prelarvae and also in the dorsal part of the urinary bladder in larvae. The expression of NKCC along the osmoregulatory epithelial cells and the presence of Na(+)/K(+)-ATPase within these cells contribute to the increase of the osmoregulatory capacity during sea bass ontogeny.


The effect of caffeine as an ergogenic aid in anaerobic exercise.
The study examined caffeine (5 mg/kg body weight) vs. placebo during anaerobic exercise. Eighteen male athletes (24.1+/-5.8 yr; BMI 26.4+/-2.2 kg/m2) completed a leg press, chest press, and Wingate test. During the caffeine trial, more total weight was lifted with the chest press, and a greater peak power was obtained during the Wingate test. No differences were observed between treatments for the leg press and average power, minimum power, and power drop (Wingate test). There was a significant treatment main effect found for postexercise glucose and insulin concentrations; higher concentrations were found in the caffeine trial. A significant interaction effect (treatment and time) was found for cortisol and glucose concentrations; both increased with caffeine and decreased with placebo. Postexercise systolic blood pressure was significantly higher during the caffeine trial. No differences were found between treatments for serum free-fatty-acid concentrations, plasma lactate concentrations, serum cortisol concentrations, heart rate, and rating of perceived exertion. Thus, a moderate dose of caffeine resulted in more total weight lifted for the chest press and a greater peak power attained during the Wingate test in competitive athletes.


Measurement of blood viscosity using a pressure-scanning capillary viscometer.
A newly designed pressure-scanning capillary viscometer is extended to measure the viscosity of whole blood over a range of shear rates without the use of anticoagulants in a clinical setting. In the present study, a single measurement of pressure variation with time replaces the flow rate and pressure drop measurements that are usually required for the operation of a capillary tube viscometer. Using a pressure transducer and capillary, we measured the variation of pressure flowing through capillary tube with respect to time, p(t), from which viscosity and the shear rate were mathematically calculated. For water and anticoagulant-added bloods, there was an excellent agreement found between the results from the pressure scanning capillary viscometer and those from a commercially available rotating viscometer. Also, the pressure-scanning capillary viscometer measured the viscosity of whole blood without heparin or EDTA. This new method overcomes the drawbacks of conventional viscometers in the measurement of whole blood viscosity. First, the pressure-scanning capillary viscometer can accurately and consistently measure the whole blood viscosity over a range of shear rates in less than 2 min without any anticoagulants. Second, this design provides simplicity (i.e., ease of operation, no moving parts, and disposable) and low cost.


Energy barrier at the N719-dye/CsSnI₃ interface for photogenerated holes in dye-sensitized solar cells.
This report is to address the question if black γ-polymorph of cesium tin tri-iodide (B-γ-CsSnI3) can be used as a solid-state hole-transport material in the conventional DSSCs with the N719 dye to replace the liquid electrolyte as reported by I. Chung et al. on Nature 485, 486, (2012). Here we demonstrate rigorously that B-γ-CsSnI3 is not energetically possible to collect photogenerated holes because of the large energy barrier at the interface of N719/B-γ-CsSnI3. Therefore, it cannot serve as a hole-transporter for the conventional DSSCs although it is a good hole-conducting material. A solution-based method was employed to synthesize the B-γ-CsSnI3 polycrystalline thin-films used for this work. These thin-films were then characterized by X-ray diffraction, Hall measurements, optical reflection, and photoluminescence (PL). Particularly, spatially resolved PL intensity images were taken after B-γ-CsSnI3 was incorporated in the DSSC structure to insure the material integrity. The means of ultraviolet photoemission spectroscopy (UPS) was used to reveal why B-γ-CsSnI3 could not act as the substitute of liquid electrolyte in the conventional DSSCs. For the completeness, other two related compounds, one is the yellow polymorph of CsSnI3 and other is Cs2SnI6 with tetravalent tin instead of double-valent tin in CsSnI3 were also investigated by UPS.


Photoactivation of CdSe/ZnS quantum dots embedded in silica colloids.
A study of the influence of the local environment on the light-induced luminescence enhancement of CdSe/ZnS quantum dots (QD) embedded in silica colloids that are dispersed in various solvents is presented. The photoluminescence of the embedded QD is enhanced up to a factor of ten upon photoactivation by ultraviolet or visible light. This enhancement is strongly dependent on the local environment. The thickness-dependent permeability of the silica shell covering the QD controls the influence of the solvent on the QD. If foreign ions are present the activation state is stabilized after termination of the activation, whereas in their absence the process is partially reversible. A new qualitative model for the photoactivation of QD in various environments is developed. It comprises light-induced passivation and subsequent oxidation processes. The embedded QD also retain their fluorescence quantum yield inside living cells. Moreover, they can be activated for many hours in living cells by laser radiation in the visible regime.


A prospective study on canine atopic dermatitis and food-induced allergic dermatitis in Switzerland.
Canine atopic dermatitis sensu stricto and food-induced allergic dermatitis are common canine skin conditions, which are often considered clinically undistinguishable. Several attempts have been made to describe populations of atopic dogs and determine breed predisposition but the results were often biased by the use of hospital populations as control group. The present study aims to describe a population of Swiss atopic and food-allergic dogs and to compare it with a data set representing more than 85% of all Swiss dogs. The study, which was carried out during 1 year in several practices and teaching hospital in Switzerland, describes a group of 259 allergic dogs, determines breed predisposition for atopic dermatitis and food-induced allergic dermatitis, compares the clinical signs and features of both conditions, and outlines the clinical picture of five frequently affected breeds.


Toward a better understanding of the process of disclosure events among people who stutter.
The purpose of this study was to identify components of disclosure events among people who stutter, and identify possible relations between these components in order to understand how disclosure events unfold. Twelve adults who stutter participated in semi-structured interviews focused on disclosure of stuttering. Participants were purposefully selected due to their self-reported history of disclosing stuttering. Qualitative content analysis using a grounded theory approach helped to identify relevant themes and subthemes related to the process of disclosure of stuttering. The findings describe the complex process of disclosure as being comprised of antecedents (including considerations about when and why to disclose), the disclosure event itself (including the content and form of the disclosure, most and least helpful methods of disclosure, as well as immediate listener reactions), and the perceived outcomes of the disclosure at individual, dyadic, and societal/contextual levels. These components of the process are linked and affect one another, resulting in a feedback loop. Disclosure methods are context-dependent and not mutually exclusive within individuals who stutter. Professionals and advocates gaining a more nuanced understanding of the process of disclosure events can increase their ability to help people who stutter make optimal decisions about disclosure. Making good disclosure decisions can help PWS improve their quality of life and reduce a variety of environmental communicative barriers.


Integrated Management Systems (IMS) to Support and Sustain Quality One Health Services: International Lessons from the COVID-19 Pandemic by the IMIA Primary Care Working Group.
One Health considers human, animal and environment health as a continuum. The COVID-19 pandemic started with the leap of a virus from animals to humans. Integrated management systems (IMS) should provide a coherent management framework, to meet reporting requirements and support care delivery. We report IMS deployment during, and retention post the COVID-19 pandemic, and exemplar One Health use cases. Six volunteer members of the International Medical Association's (IMIA) Primary Care Working Group provided data about any IMS and One Health use to support the COVID-19 pandemic initiatives. We explored how IMS were: (1) Integrated with organisational strategy; (2) Utilised standardised processes, and (3) Met reporting requirements, including public health. Selected contributors provided Unified Modelling Language (UML) use case diagram for a One Health exemplar. There was weak evidence of synergy between IMS and health system strategy to the COVID-19 pandemic. However, there were rapid pragmatic responses to COVID-19, not citing IMS. All health systems implemented IMS to link COVID test results, vaccine uptake and outcomes, particularly mortality and to provide patients access to test results and vaccination certification. Neither proportion of gross domestic product alone, nor vaccine uptake determined outcome. One Health exemplars demonstrated that animal, human and environmental specialists could collaborate. IMS use improved the pandemic response. However, IMS use was pragmatic rather than utilising an international standard, with some of their benefits lost post-pandemic. Health systems should incorporate IMS that enables One Health approaches as part of their post COVID-19 pandemic preparedness.


Bilirubin level fluctuation in drain discharge after hepatectomies justifies long-term drain maintenance.
Need for abdominal drains after liver resection is debated. However, unrecognized bile leak is relatively frequent: to prevent bile collection we adopted the use of long-term drains. The aim of this study was to validate this policy checking the bilirubin concentration in the drain discharge and serum along the postoperative course. A prospective cohort study enrolling 58 consecutive patients with liver tumors was carried out. All patients underwent liver resection and received abdominal drains which were maintained for at least 7 days postoperatively. The bilirubin concentration in serum and drain discharge was sampled on the 3rd, 5th and 7th postoperative days. No postoperative mortality and major morbidity were observed. The bilirubin level in drain discharge was higher on the 5th postoperative day than on the 3rd and 7th postoperative days: difference between the 3rd and 5th postoperative days was significant. No differences were observed among serum bilirubin levels on 3rd, 5th and 7th postoperative days. The bilirubin level in drain discharge increases late in the postoperative course. Therefore, bile leakage should be evaluated between the 5th and 7th postoperative days. The use of long-term drains helps protect against undiscovered collections and thus impacts postoperative course.


A new lipoglycopeptide: telavancin.
The increase in infections caused by resistant Gram-positive organisms has led to an urgent need for new antibiotics. Telavancin is a rapidly bactericidal lipoglycopeptide with multiple mechanisms of action, including concentration-dependent inhibition of bacterial cell wall synthesis and disruption of the functional integrity of the cell membrane. Telavancin is active against a wide variety of Gram-positive organisms including Staphylococcus aureus with resistance to methicillin, reduced susceptibility to vancomycin, and full resistance to vancomycin. Telavacin is approximately 90% protein bound; it has a serum half-life of around 8 h and a prolonged postantibiotic effect, allowing once daily administration. Telavancin is eliminated principally through the urine, requiring dose adjustment in patients with renal impairment. The efficacy and safety of telavancin was demonstrated in a large program of patients with complicated skin and skin structure infections. Development of resistance has not been detected in clinical strains. Adverse events include taste disturbance, nausea and vomiting; a small proportion of patients experienced reversible increase in serum creatinine. Two large Phase III studies in patients with healthcare associated pneumonia were recently completed. Telavancin has the potential to become an important therapeutic option to treat serious infections produced by resistant Gram-positive cocci, particularly those caused by methicillin-resistant S. aureus.


Quinolone-resistant Campylobacter infections: risk factors and clinical consequences.
We integrated data on quinolone and macrolide susceptibility patterns with epidemiologic and typing data from Campylobacter jejuni and C. coli infections in two Danish counties. The mean duration of illness was longer for 86 patients with quinolone-resistant C. jejuni infections (median 13.2 days) than for 381 patients with quinolone-sensitive C. jejuni infections (median 10.3 days, p = 0.001). Foreign travel, eating fresh poultry other than chicken and turkey, and swimming were associated with increased risk for quinolone-resistant C. jejuni infection. Eating fresh chicken (of presumably Danish origin) was associated with a decreased risk. Typing data showed an association between strains from retail food products and broiler chickens and quinolone-sensitive domestically acquired C. jejuni infections. An association between treatment with a fluoroquinolone before stool-specimen collection and having a quinolone-resistant C. jejuni infection was not observed.


Cardiac autonomic nervous system and risk of arrhythmias in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited systemic microangiopathy with prevalently cerebral manifestations. Among the causes of death, sudden unexpected death seems to occur in a significant number of CADASIL patients. Because potential causes of sudden unexpected death may include cardiac arrhythmias and myocardial infarction, we evaluated risk factors for life-threatening arrhythmias, such as reduced heart rate variability, sympathetic overactivity and QT interval (QTc) prolongation, in 23 CADASIL patients. The relationship of these changes with brain MRI pattern was also investigated. Frequency domain measures of heart rate variability (10 minutes recordings) and QTc interval were recorded in 23 CADASIL patients (17 males, 6 females) and 22 healthy age- and sex-matched control subjects. The following heart rate variability spectral parameters were considered at rest during spontaneous and controlled breathing (Cb): total power, very-low-frequency component, low-frequency component, high-frequency component, low-frequency/high-frequency ratio, and Cb-total power, Cb-very-low-frequency component, Cb-low-frequency component, Cb-high-frequency component, Cb-low-frequency/high-frequency ratio. R-to-R wave and QTc interval were also analyzed. All data were statistically compared between CADASIL and control subjects. Conventional brain MRI was performed in patients with CADASIL and T1-weighted and T2-weighted lesion volumes, and were compared with each spectral component of the tachogram. During spontaneous and controlled breathing, total power spectrum and all spectral components (very low frequency component, high-frequency component, low-frequency component) of heart rate variability were significantly reduced in CADASIL patients with respect to controls (P<0.05). The low-frequency/high-frequency component ratio was significantly higher in CADASIL patients than in controls. No significant correlation between heart rate variability spectral parameters and other variables including total brain T2-weighted and T1-weighted lesion volumes were observed in CADASIL subjects. We found a statistically significant reduction in all frequency domain parameters of heart rate variability associated with a higher low frequency/high frequency ratio for CADASIL patients with respect to normal subjects. These data are consistent with autonomic derangement and suggests that CADASIL patients may be at risk for life-threatening arrhythmias. This could at least in part explain their higher recurrence of sudden unexpected death and should be taken into account in planning therapy.


Nonclassical Islet Peptides: Pancreatic and Extrapancreatic Actions.
The pancreas has physiologically important endocrine and exocrine functions; secreting enzymes into the small intestine to aid digestion and releasing multiple peptide hormones via the islets of Langerhans to regulate glucose metabolism, respectively. Insulin and glucagon, in combination with ghrelin, pancreatic polypeptide and somatostatin, are the main classical islet peptides critical for the maintenance of blood glucose. However, pancreatic islets also synthesis numerous 'nonclassical' peptides that have recently been demonstrated to exert fundamental effects on overall islet function and metabolism. As such, insights into the physiological relevance of these nonclassical peptides have shown impact on glucose metabolism, insulin action, cell survival, weight loss, and energy expenditure. This review will focus on the role of individual nonclassical islet peptides to stimulate pancreatic islet secretions as well as regulate metabolism. In addition, the more recognised actions of these peptides on satiety and energy regulation will also be considered. Furthermore, recent advances in the field of peptide therapeutics and obesity-diabetes have focused on the benefits of simultaneously targeting several hormone receptor signalling cascades. The potential for nonclassical islet hormones within such combinational approaches will also be discussed.


Mixtures of endocrine-disrupting contaminants induce adverse developmental effects in preweaning rats.
Reproductive toxicity was investigated in rats after developmental exposure to a mixture of 13 endocrine-disrupting contaminants, including pesticides, plastic and cosmetic ingredients, and paracetamol. The mixture was composed on the basis of information about high-end human exposures, and the dose levels reflecting 100, 200, and 450 times this exposure were tested. The compounds were also grouped according to their estrogenicity or anti-androgenicity, and their joint effects were tested at two different doses, with each group reflecting 200 or 450 times human exposure. In addition, a single paracetamol dose was tested (350 mg/kg per day). All exposures and a vehicle were administered by oral gavage to time-mated Wistar dams rats throughout gestation and lactation, and their offspring were assessed for reproductive effects at birth and in prepuberty. The mixture doses, which included the anti-androgenic compounds, affected the male offspring by causing decreased anogenital distance, increased nipple retention (NR), and reduced ventral prostate weights, at both medium and high doses. In addition, the weights of the levator ani/bulbocavernosus muscle (LABC) were decreased at the high dose of anti-androgen mixture. No effects were seen after exposure to the estrogenic chemicals alone, whereas males exposed solely to paracetamol showed decreased LABC weights and increased NR. Thus adverse reproductive effects were observed at mixtures reflecting 200 times high-end human exposure, which is relatively close to the safety margin covered by the regulatory uncertainty factor of 100. This suggests that highly exposed human population groups may not be sufficiently protected against mixtures of endocrine-disrupting chemicals.


0-7-21 hypofractionated palliative radiotherapy: an effective treatment for advanced head and neck cancers.
We report our experience in providing palliative radiotherapy (RT) to patients with head and neck cancers (HNCs). Our hypofractionated regimen, "0-7-21", treats patients with 24 Gy in three fractions. Patients, disease and response data were retrieved for candidates of 0-7-21 from 2005 to 2012. Primary end points included symptom and tumour size responses to RT based on response evaluation criteria in solid tumours (RECIST) guidelines. Secondary end points included progression-free survival (PFS) within the irradiated field, overall survival (OS) and symptomatic PFS (SPFS), calculated using Kaplan-Meier method and adverse events. Cox proportional hazards regression and logistic regression were used to investigate for prognostic factors. A total of 110 patients were included. Among the patients, 40% and 31% had complete response for symptoms and tumour size, respectively; 42% and 50% had partial response for symptoms and tumour size, respectively; and 15% had stability of symptoms and tumour size. Median 6-month OS was 51%, and PFS within the irradiated field was 39%. Planning target volume was predictive of OS (p < 0.001), PFS (p < 0.001) and SPFS (p < 0.005), while higher TNM stage was associated with poorer tumour response (p = 0.02). 0-7-21 is an effective and well-tolerated palliative RT regimen for patients with HNC. There was excellent symptom and local control with acceptable toxicity profile in these patients. This is the first study to describe the outcomes of 0-7-21 in treating advanced HNCs. The positive results suggest that 0-7-21 provides excellent palliation with minimal toxicity, with significantly less on-treatment time than current published palliative RT regimen.


Food matrix and processing influence on carotenoid bioaccessibility and lipophilic antioxidant activity of fruit juice-based beverages.
The biological activity of carotenoids depends on their bioaccessibility and solubilization in the gastrointestinal tract. These compounds are poorly dispersed in the aqueous media of the digestive tract due to their lipophilic nature. Thus, it is important to analyze the extent to which some factors, such as the food matrix and food processing, may improve their bioaccessibility. Beverages formulated with a blend of fruit juices and water (WB), milk (MB) or soymilk (SB) were treated by high-intensity pulsed electric fields (HIPEF) (35 kV cm(-1) with 4 μs bipolar pulses at 200 Hz for 1800 μs), high-pressure processing (HPP) (400 MPa at 40 °C for 5 min) or thermal treatment (TT) (90 °C for 1 min) in order to evaluate the influence of food matrix and processing on the bioaccessibility of carotenoids and on the lipophilic antioxidant activity (LAA). The bioaccessibility of these compounds diminished after applying any treatment (HIPEF, HPP and TT), with the exception of cis-violaxanthin + neoxanthin, which increased by 79% in HIPEF and HPP beverages. The lowest carotenoid bioaccessibility was always obtained in TT beverages (losses up to 63%). MB was the best food matrix for improving the bioaccessibility of carotenoids, as well as the LAA. The results demonstrate that treatment and food matrix modulated the bioaccessibility of carotenoids as well as the lipophilic antioxidant potential of beverages. Additionally, HIPEF and HPP could be considered as promising technologies to obtain highly nutritional and functional beverages.


Marijuana intoxication in a cat.
Cannabis from hemp (Cannabis sativa and C. indica) is one of the most common illegal drugs used by drug abusers. Indian cannabis contains around 70 alkaloids, and delta-9-tetrahydrocannabinol (delta-9-THC) is the most psychoactive substance. Animal intoxications occur rarely and are mostly accidental. According to the US Animal Poison Control Center, cannabis intoxication mostly affects dogs (96%). The most common cause of such intoxication is unintentional ingestion of a cannabis product, but it may also occur after the exposure to marijuana smoke. A 6-year-old Persian cat was brought to the veterinary clinic due to strong psychomotor agitation turning into aggression. During hospitalisation for 14 days, the cat behaved normally and had no further attacks of unwanted behaviour. It was returned to its home but shortly after it developed neurological signs again and was re-hospitalised. On presentation, the patient showed no neurological abnormalities except for symmetric mydriasis and scleral congestion. During the examination, the behaviour of the cat changed dramatically. It developed alternate states of agitation and apathy, each lasting several minutes. On interview it turned out that the cat had been exposed to marijuana smoke. Blood toxicology tests by gas chromatography tandem mass spectrometry revealed the presence of delta-9-tetrahydrocannabinol (THC) at 5.5 ng/mL, 11-hydroxy-delta-9-THC at 1.2 ng/mL, and 11-carboxy-delta-9-THC at 13.8 ng/mL. The cat was given an isotonic solution of NaCl 2.5 and 2.5% glucose at a dose of 40 mL/kg/day parenterally and was hospitalised. After complete recovery, the cat was returned to it's owner and future isolation of the animal from marijuana smoke was advised. This is the first case of a delta-9-tetrahydrocannabinol intoxication in a cat with both description of the clinical findings and the blood concentration of delta-9-THC and its main metabolites.


Adenosine is not responsible for local metabolic control of coronary blood flow in dogs during exercise.
The purpose of this investigation was to quantitatively evaluate the role of adenosine in coronary exercise hyperemia. Dogs (n = 10) were chronically instrumented with catheters in the aorta and coronary sinus, and a flow probe on the circumflex coronary artery. Cardiac interstitial adenosine concentration was estimated from arterial and coronary venous plasma concentrations using a previously tested mathematical model. Coronary blood flow, myocardial oxygen consumption, heart rate, and aortic pressure were measured at rest and during graded treadmill exercise with and without adenosine receptor blockade with either 8-phenyltheophylline (8-PT) or 8-p-sulfophenyltheophylline (8-PST). In control vehicle dogs, exercise increased myocardial oxygen consumption 4.2-fold, coronary blood flow 3.8-fold, and heart rate 2.5-fold, whereas mean aortic pressure was unchanged. Coronary venous plasma adenosine concentration was little changed with exercise, and the estimated interstitial adenosine concentration remained well below the threshold for coronary vasodilation. Adenosine receptor blockade did not significantly alter myocardial oxygen consumption or coronary blood flow at rest or during exercise. Coronary venous and estimated interstitial adenosine concentration did not increase to overcome the receptor blockade with either 8-PT or 8-PST as would be predicted if adenosine were part of a high-gain, negative-feedback, local metabolic control mechanism. These results demonstrate that adenosine is not responsible for local metabolic control of coronary blood flow in dogs during exercise.


Relationship between bond strengths and nanoleakage: evaluation of a new assessment method.
To test if there was a correlation between resin-dentin bond strengths and nanoleakage of total etch and self-etching primer bonding systems. Ten extracted third molars were ground flat with 600 grit SIC paper under running water to expose middle dentin. Clearfil Liner Bond 2V or Single Bond was applied to the dentin surface according to the manufacturers' instructions. A crown was built-up with Clearfil AP-X resin-based composite, and the specimens stored in water for 24 hrs at 37 degrees C. The bonded assemblies were cut mesiodistally perpendicular to the interface into approximately 0.7 mm thick slabs, and trimmed for microtensile bond strength testing. Alternate slices from each tooth were either entirely double-coated with nail varnish leaving a 0.5 mm window around the interface, or left uncoated to be immersed in water for 13 hrs as the control. The varnished (experimental) slabs were immersed in 50% AgNO3 for 1 hr, and then immersed in a photo developing solution for 12 hrs. All specimens were subjected to microtensile bond test at a crosshead speed of 1 mm/min. After debonding, the specimens were embedded in epoxy resin and observed under a confocal laser scanning microscope for determination of lateral silver penetration within the interface. Silver penetration was further analyzed under the SEM. Data were analyzed with one- and two-way ANOVA, Fisher's PLSD test (P < 0.05), and regression analysis. The bond strengths of Clearfil Liner Bond 2V and Single Bond were not significantly different (P > 0.05); however the bond strengths of the specimens that were soaked in 50% AgNO3 for 1 hr significantly increased (P < 0.01). No correlation was observed between bond strengths and nanoleakage.


Mechanisms of cancer prevention by tea constituents.
Consumption of tea (Camellia sinensis) has been suggested to prevent cancer, heart disease and other diseases. Animal studies have shown that tea and tea constituents inhibit carcinogenesis of the skin, lung, oral cavity, esophagus, stomach, liver, prostate and other organs. In some studies, the inhibition correlated with an increase in tumor cell apoptosis and a decrease in cell proliferation. Studies with human cancer cell lines have demonstrated that epigallocatechin-3-gallate (EGCG), a major tea polyphenol, inhibits mitogen-activated protein kinases, cyclin-dependent kinases, growth factor-related cell signaling, activation of activator protein 1 (AP-1) and nuclear factor kappaB (NFkappaB), topoisomerase I and matrix metalloproteinases as well as other potential targets. Although some studies report effects of EGCG at submicromolar levels, most experiments require concentrations of >10 or 20 micromol/L to demonstrate the effect. In humans, tea polyphenols undergo glucuronidation, sulfation, methylation, and ring fission. The peak plasma concentration of EGCG is approximately 1 micromol/L. The possible relevance of each of the proposed mechanisms to human cancer prevention is discussed in light of current bioavailability data for tea polyphenols and the potential limitations of animal models of carcinogenesis. Such discussion, it is hoped, will clarify some misunderstandings of cancer prevention by tea and stimulate new research efforts.


The interference of paid work with household demands in different social policy contexts: perceived work-household conflict in Sweden, the UK, the Netherlands, Hungary, and the Czech Republic.
The article explores whether people experiences a lower level of work-household conflict in a context that is characterized by extensive family policies (Sweden and to some extent Hungary and Czech Republic) aimed at facilitating participation in the labour market. This is done by studying perceived work-household conflict among women and men living in Sweden, the UK, the Netherlands, Hungary, and the Czech Republic. The analyses are based on the answers to a questionnaire distributed to nearly 6,000 randomly selected individuals within the framework of the European Union financed 'Household, Work, and Flexibility' (HWF) study. The results show that women in Sweden experience conflicts between work and household demands to a higher degree than any other category in all five countries. The differences between Swedish women and women living in the Netherlands and the UK are explained by variables indicating qualifications and workload in the main job, but the lower degree of work-household conflict among Czech and Hungarian women is still significant when controlling for household composition and working conditions. Data indicate that a possible explanation for this can be found in the interplay between men's and women's attitudes toward gender roles and the actual situation in terms of division of labour.


Residual air persists in the renal collecting system following percutaneous nephrolithotomy.
To determine the presence or absence of air in the renal collecting system 48 hours following percutaneous nephrolithotomy (PCNL). The study was based on a review of the records of the last 100 consecutive patients undergoing PCNL for stone extraction. Two urologists and a uroradiologist reviewed post-PCNL nephrostogram films and determined the presence or absence of air in the urinary tract. Nephrostography was routinely performed 48 hours after PCNL, and all films of the 100 patients were reviewed. Gas was detected in 98 (98%) of the 100 cases that appeared as gas pyelograms outlining the renal calyces. Of the 98 cases, 33 (34%) demonstrated gas in the upper calyx, 10 (10%) in the middle calyx, 12 (13%) in the lower calyx, and 43 (44%) in more than one calyx. Gas was not detected in the renal parenchyma or perirenal areas. Gas is present within the renal collecting system 48 hours after a percutaneous procedure. Prospective studies should be performed to document the amount of time needed for complete resolution of gas from the collecting system.


Camel Streptococcus agalactiae populations are associated with specific disease complexes and acquired the tetracycline resistance gene tetM via a Tn916-like element.
Camels are the most valuable livestock species in the Horn of Africa and play a pivotal role in the nutritional sustainability for millions of people. Their health status is therefore of utmost importance for the people living in this region. Streptococcus agalactiae, a Group B Streptococcus (GBS), is an important camel pathogen. Here we present the first epidemiological study based on genetic and phenotypic data from African camel derived GBS. Ninety-two GBS were characterized using multilocus sequence typing (MLST), capsular polysaccharide typing and in vitro antimicrobial susceptibility testing. We analysed the GBS using Bayesian linkage, phylogenetic and minimum spanning tree analyses and compared them with human GBS from East Africa in order to investigate the level of genetic exchange between GBS populations in the region. Camel GBS sequence types (STs) were distinct from other STs reported so far. We mapped specific STs and capsular types to major disease complexes caused by GBS. Widespread resistance (34%) to tetracycline was associated with acquisition of the tetM gene that is carried on a Tn916-like element, and observed primarily among GBS isolated from mastitis. The presence of tetM within different MLST clades suggests acquisition on multiple occasions. Wound infections and mastitis in camels associated with GBS are widespread and should ideally be treated with antimicrobials other than tetracycline in East Africa.


Water circulation and governing factors in humid tropical river basins in the central Western Ghats, Karnataka, India.
The small river basins in the narrow stretch of the Arabian Sea coast of southwest India experience high annual rainfall (800-8000 mm), with a higher proportion (85 %) during the summer monsoon period between June and September. This is due to a unique orographic barrier provided by the Western Ghats mountain belt (600-2600 m) for the summer monsoon brought by the southwesterly winds. This study is the first of a kind focusing on the water cycle with an intensive stable isotopes approach (samples of river water, groundwater, rainwater; seasonal and spatial sampling) in this part of the Western Ghats in Karnataka and also in the highest rainfall-receiving region (with places like Agumbe receiving 7000-8000 mm annual rainfall) in South India. In addition, the region lacks sustainable water budgeting due to high demographic pressure and a dry pre-monsoon season as the monsoon is mainly unimodal in this part of India, particularly close to the coast. The stable isotopic compositions of groundwater, river water and rainwater in two tropical river basins situated approximately 60 km apart, namely the Swarna near Udupi and the Nethravati near Mangalore, were studied from 2010 to 2013. The δ(18)O and δ(2)H values of the water samples were measured by isotope ratio mass spectrometry, and the d-excess values calculated to better understand the dominant source of the water and the influence of evaporation/recycling processes. The water in the smaller area basin (Swarna basin) does not show seasonal variability in the δ(18)O values for groundwater and river water, having a similar mean value of -3.1 ‰. The d-excess value remains higher in both wet and dry seasons suggesting strong water vapor recycling along the foothills of the Western Ghats. In contrast, the larger tropical basin (Nethravati basin) displays specific seasonal isotopic variability. The observation of higher d-excess values in winter with lower δ(18)O values suggests an influence of northeast winter monsoon water in the larger basin. The narrow coastal strip to the west of the Western Ghats displays unique water characteristics in both tropical river basins investigated. For the smaller and hilly Swarna basin, the dense vegetation (wet canopies) could largely re-evaporate the (intercepted) rain, leading to no marked seasonal or altitude effect on the water isotope values within the basin. The larger Nethravati basin, which stretches farther into the foothills of the Western Ghats, receives winter monsoon water, and thus exhibits a clear seasonal variability in rainfall moisture sources. The degree of water vapor recycling in these wet tropical basins dominates the isotopic composition in this narrow coastal stretch of South India. An insight into the soil water contribution to the river water and groundwater, even in the rainfall-dependent tropical basins of South India, is provided in this study.


Sensitivity of point-of-care testing C reactive protein and procalcitonin to diagnose urinary tract infections in Dutch nursing homes: PROGRESS study protocol.
Suspected urinary tract infection (UTI) ranks among the most common reasons for antibiotic use in nursing homes. However, diagnosing UTI in this setting is challenging because UTI often presents with non-specific symptomatology. Moreover asymptomatic bacteriuria is common in elderly, which complicates attribution of causality to detection of bacteria in urine. These diagnostic challenges contribute to overuse of antibiotics and emergence of antimicrobial resistance in nursing homes. Given the diagnostic challenges, there is a need for point-of-care (POC) diagnostic tests to support clinical rules for diagnosing UTI. Procalcitonin (PCT) and C reactive protein (CRP) are inflammatory blood markers that have been proven useful to support diagnosis and monitoring of (bacterial) respiratory tract infections and sepsis. While limited studies suggest their usefulness in supporting UTI diagnosis, their utility has not been studied in elderly populations for this purpose. In a 24-month matched prospective study, 'PROGRESS' will assess and compare the sensitivity of rapid POC measurements of blood CRP and PCT levels to support clinical rules for diagnosing UTI in nursing home residents. The primary outcome measure is sensitivity of the POC tests to identify patients with true UTI based on the predefined definition, as derived from receiver operating curves. This study will be conducted in accordance with Good Clinical Practice guidelines and the principles of the Declaration of Helsinki. The study protocol is approved by the Medical Ethical Committee of Amsterdam UMC location VUmc with reference number 2017.350 and National Central Committee on Research involving Human Subjects with reference number NL62067.029.17. NTR6467.


Thermal and pasting properties and digestibility of blends of potato and rice starches differing in amylose content.
In this study, waxy or normal potato starches (WPS or NPS) were blended with waxy, low-amylose or high-amylose rice starch (WRS, LARS or HARS) in different ratios (100:0, 80:20, 60:40, 40:60, 20:80 and 0:100). Pasting profiles of blends were additively between those of the component starches separately except for some mixtures of WRS and potato starches. Twin or even three gelatinization peaks were observed for potato-WRS or potato-HARS blend, while only one peak was observed in potato-LARS blend. Addition of WRS enhanced the nutritional profile of blends containing WPS by decreasing the rapidly digestible starch level. Microscopy revealed that addition of WRS was beneficial for the development of dense and compact structure of gels of blends compared with their counterparts, which may be because few amylose chains leached to inhibit the interaction between swollen potato and rice starch granules. Besides, peak, trough, breakdown and final viscosity as well as gelatinization enthalpy showed significantly negative correlations with amylose leaching. Non-additive behaviours were observed for properties, but more independent behaviour was observed between potato starch and LARS or HARS. Results suggested that properties of blends of potato and rice starches differing in amylose content varied through different extents of amylose leaching.


Ethylene signaling pathway and MAPK cascades are required for AAL toxin-induced programmed cell death.
Programmed cell death (PCD), known as hypersensitive response cell death, has an important role in plant defense response. The signaling pathway of PCD remains unknown. We employed AAL toxin and Nicotiana umbratica to analysis plant PCD. AAL toxin is a pathogenicity factor of the necrotrophic pathogen Alternaria alternata f. sp. lycopersici. N. umbratica is sensitive to AAL toxin, susceptible to pathogens, and effective in Tobacco rattle virus-based virus-induced gene silencing (VIGS). VIGS analyses indicated that AAL toxin-triggered cell death (ACD) is dependent upon the mitogen-activated protein (MAP) kinase kinase MEK2, which is upstream of both salicylic acid-induced protein kinase (SIPK) and wound-induced protein kinase (WIPK) responsible for ethylene (ET) synthesis. ET treatment of MEK2-silenced N. umbratica re-established ACD. In SIPK- and WIPK-silenced N. umbratica, ACD was compromised and ET accumulation was not observed. However, in contrast to the case of MEK2-silenced plants, ET treatment did not induce cell death in SIPK- and WIPK-silenced plants. This work showed that ET-dependent pathway and MAP kinase cascades are required in ACD. Our results suggested that MEK2-SIPK/WIPK cascades have roles in ET biosynthesis; however, SIPK and WIPK have other roles in ET signaling or another pathway leading to cell death by AAL toxin.


Phasic responses in dorsal raphe serotonin neurons to noxious stimuli.
Serotonin is widely implicated in aversive processing. It is not clear, however, whether serotonin neurons encode information about aversive stimuli. We found that, in the dorsal raphe of anesthetized rats, most neurochemically-identified clocklike serotonin neurons were phasically excited by noxious footshocks, whereas most bursting serotonin neurons were inhibited. These results suggest that discrete groups of serotonin neurons differentially code for aversive stimuli.


The relationship between inpatient hyperglycaemia and mortality is modified by baseline glycaemic status.
There is a well-established association between inpatient hyperglycaemia and mortality. However, evidence is inconsistent regarding whether this association is differential among those with and without type 2 diabetes mellitus (T2DM). Most studies are based on convenience samples or are unable to adjust for comorbidities. We examined whether the association between hyperglycaemia and 30-day mortality was modified by baseline glycaemic status. This was a retrospective cohort study of 174,671 eligible hospitalized individuals between 2012 and 2015. Thirty-day mortality was assessed during the first inpatient stay up to 30 days post discharge. The adjusted association between hyperglycaemia and mortality was assessed with logistic regression models. Then, four interaction terms were entered into the model to assess if the association between hyperglycaemia and mortality differed by baseline glycaemic status. The multivariate model demonstrated a 2.18-fold risk of mortality associated with hyperglycaemia (odds ratio [OR] [95%CI]: 2.19 [2.08-2.31]). Adding the interaction terms between hyperglycaemia and baseline glycaemic status the ORs of 30-day mortality were 1.41 (1.25-1.60) in non-T2DM status, 1.32 (1.16-1.51) in pre-diabetes status and 1.30 (1.04-1.62) in unscreened status, as compared to T2DM status with hyperglycaemia. Hyperglycaemia is positively associated with mortality and both those without and with controlled T2DM are at highest risk. These findings may help medical staff identify potential increased risk of mortality upon hospital entry and discharge, and direct further research to assess how hyperglycaemia control and proactive deterioration prevention throughout the entire inpatient stay may prevent adverse outcomes.


Effects of Melanocortin 3 and 4 Receptor Deficiency on Energy Homeostasis in Rats.
Melanocortin-3 and 4 receptors (MC3R and MC4R) can regulate energy homeostasis, but their respective roles especially the functions of MC3R need more exploration. Here Mc3r and Mc4r single and double knockout (DKO) rats were generated using CRISPR-Cas9 system. Metabolic phenotypes were examined and data were compared systematically. Mc3r KO rats displayed hypophagia and decreased body weight, while Mc4r KO and DKO exhibited hyperphagia and increased body weight. All three mutants showed increased white adipose tissue mass and adipocyte size. Interestingly, although Mc3r KO did not show a significant elevation in lipids as seen in Mc4r KO, DKO displayed even higher lipid levels than Mc4r KO. DKO also showed more severe glucose intolerance and hyperglycaemia than Mc4r KO. These data demonstrated MC3R deficiency caused a reduction of food intake and body weight, whereas at the same time exhibited additive effects on top of MC4R deficiency on lipid and glucose metabolism. This is the first phenotypic analysis and systematic comparison of Mc3r KO, Mc4r KO and DKO rats on a homogenous genetic background. These mutant rats will be important in defining the complicated signalling pathways of MC3R and MC4R. Both Mc4r KO and DKO are good models for obesity and diabetes research.


An epidemiological study of acute carbon monoxide poisoning in the West Midlands.
To describe the epidemiology of carbon monoxide (CO) poisoning in a defined population, identifying those at greatest risk from acute poisoning resulting in admission to hospital or death. A retrospective study with routinely collected information, set in the former West Midlands Regional Health Authority; population of 5.2 million. The data comprised 939 deaths and 701 hospital admissions due to CO poisoning between January 1988 to December 1994. The main outcome measures were age and sex standardised incidence rates (SIRs) for non-intentional, suicidal, and undetermined poisonings for health authorities and the linear relation with socioeconomic deprivation. Overall rate of non-intentional poisonings over the 7 year period was 7.6/100,000, an annual rate of 1.1/100,000. The 7 year rates were highest in people > or = 85; men 24.0/100,000 and women 19.7/100,000. For suicides the 7 year rate was 19.6/100,000, an annual rate of 2.8/100,000. The 7 year rates were highest for men of 35-39, 64.1/100,000, and for women aged 45-49, 15.3/100,000. None of the causes of poisoning were related to deprivation. Non-intentional poisonings showed a strong seasonal variation with the highest rates being recorded in the months October to March. Increased rates of poisoning were found in the rural districts of the West Midlands. There seems to have been a decline in suicides coinciding with the introduction of three way catalytic converters on cars. Elderly people and the very young are at the greatest risk from non-intentional CO poisoning and rates are highest in the winter months. Although deaths from non-intentional CO poisoning are declining nationally, in the West Midlands they have remained stable and hospital admissions are increasing. It is not solely an urban phenomenon with rates for non-intentional CO poisoning and suicides higher in the rural districts. Health authorities need to consider all populations in any prevention programme. Further work is needed to establish the extent of the burden of chronic CO poisoning and the impact of catalytic converters on suicides.


International differences in the management of intracranial aneurysms: implications for the education of the next generation of neurosurgeons.
The publication of the International Subarachnoid Aneurysm Trial rapidly changed the management of patients with subarachnoid hemorrhage. The present and perceived future trends of aneurysm management have significant implications for patients and how we educate future cerebrovascular specialists. To determine present perceived competencies of final-year neurosurgical residents who have just finished their residencies and to relate those to what practitioners from a variety of continents expect of these persons. The goal is to provide a basis for further discussion regarding the design of further educational programs in neurosurgery. A 55-item questionnaire with 33 questions related to competencies and expectations of competency from final-year residents who have just finished residency was completed by 229 neurosurgeons and neuro-radiologists (81 % response rate) of mixed seniority from 45 countries. We used bivariate and descriptive analyses to determine future trends and geographic differences in cerebral aneurysm management as well as the educational implications on the future. More North Americans than those from the rest of the world are of the opinion that graduating residents are presently competent to perform basic cerebrovascular procedures like evacuation of a hematoma and clipping a simple 7-mm middle cerebral artery aneurysm. Extremely few graduating neurosurgical residents anywhere are presently capable of performing endovascular techniques for even the most basic of aneurysms. Most of those surveyed also believe that endovascular and open surgical management of aneurysms should be a part of residency training for all residents (70.4 and 88.7 %, respectively). Our findings have implications for the design of neurosurgical curricula for residents as well as for certification examinations and procedures. Specialty and educational organizations and those responsible for the education of future clinicians who will care for patients with cerebrovascular problems should adjust educational objectives and implement curricula and learning experiences that will ensure that cerebrovascular specialists are capable of providing the best care possible to the patient with an aneurysm, whether that be open surgery or endovascular management. These findings mean that organizations around the world will need to make these adjustments to the education of future specialists.


Efficacy of intravaginal 0.75% metronidazole gel for the treatment of bacterial vaginosis.
To assess the efficacy of 0.75% metronidazole vaginal gel, formulated at pH 4.0, for the treatment of bacterial vaginosis in a double-blind, placebo-controlled crossover trial. Fifty-three women with bacterial vaginosis (clue cells of at least 20% plus vaginal pH of at least 4.7, amine odor, or homogeneous discharge) were enrolled. Women with concomitant infection by Candida, Trichomonas, Chlamydia trachomatis, or Neisseria gonorrhoeae were excluded. Five grams of metronidazole gel or placebo was inserted intravaginally twice daily for 5 days. Response to therapy was evaluated at 4-16 days and 1 month after treatment. Twenty-six of the 30 women (87%) who received metronidazole were free of bacterial vaginosis 9-21 days after enrollment, compared with only four (17%) of 23 placebo-treated women (P < .001). Women who were cured were followed prospectively. Those who had persistent bacterial vaginosis were crossed over to metronidazole gel in an open trial; bacterial vaginosis was eradicated in 21 of 23 (91%) of these women at the 2-week follow-up. Recurrent bacterial vaginosis developed within 1 month in four (15%) of 26 women who were initially cured after receiving metronidazole gel. Response to therapy was accompanied by a decrease in vaginal Bacteroides, Gardnerella vaginalis, and Mycoplasma hominis. Treatment with intravaginal metronidazole gel resulted in a clinical cure in 87% (placebo-controlled trial) to 91% (crossover trial) of women with bacterial vaginosis. The recurrence rate of 15% at 1 month after treatment is similar to that reported with oral metronidazole.