Usual Clinical Practice for Early Supported Discharge after Stroke with Continued Rehabilitation at Home: An Observational Comparative Study.
Based on randomised controlled trials, evidence exists that early supported discharge (ESD) from the hospital with continued rehabilitation at home has beneficial effects after stroke; however, the effects of ESD service in regular clinical practice have not been investigated. The purpose of the current study was to compare ESD service with conventional rehabilitation in terms of patient outcomes, caregiver burden at 3 and 12 months and the use and costs of healthcare during the first year after stroke. This study was a subgroup analysis of a longitudinal observational study of patients who received care in the stroke unit at Karolinska University Hospital in Sweden. Patients who met the inclusion criteria for ESD in previous experimental studies were included. The patients were referred to available rehabilitation services at discharge, and comparisons between those who received ESD service (the ESD group, n = 40) and those who received conventional rehabilitation (the NoESD group, n = 110) were performed with regard to independence in activities of daily living (ADL), the frequency of social activities, life satisfaction, and caregiver burden and the use and costs of healthcare during the first year after stroke. At 3 and 12 months, no differences were observed with regard to patient outcomes; however, ESD was associated with a lower caregiver burden (p = 0.01) at 12 months. The initial length of stay (LOS) at the hospital was 8 days for the ESD group and 15 days for the NoESD group (p = 0.02). The median number of outpatient rehabilitation contacts was 20.5 for the ESD group (81% constituting ESD service) and 3 for the NoESD group (p<0.001). There was no difference between the groups with regard to overall healthcare costs. ESD service in usual clinical practice renders similar health benefits as conventional rehabilitation but a different pattern of resource use and with released capacity in acute stroke care.


Isolation and characterization of two H5N1 influenza viruses from swine in Jiangsu Province of China.
Pigs are susceptible to infection with both human and avian influenza A viruses and are considered intermediate hosts that facilitate virus reassortment. Although H5N1 virus has spread to a wide range of avian and mammalian species, data about swine H5N1 isolates are scarce. To determine whether Asian H5N1 influenza viruses had been transmitted to pigs, a total of 1,107 nasal swab samples from healthy swine were collected from 2008 to 2009 in Jiangsu province of eastern China. In this survey, two H5N1 viruses A/swine/Jiangsu/1/2008 (JS/08) and A/swine/Jiangsu/2/2009 (JS/09) were isolated and identified. Phylogenetic analysis showed that JS/08 and JS/09 belonged to clade 7 and clade 2.3.4, respectively, and shared over 99.0 % sequence identity with poultry H5N1 isolates of the same clade in China. Receptor specificity analysis also showed that both of the swine H5N1 isolates bound preferentially to avian-type receptors. However, experiments in mammals indicated that JS/09 was moderately pathogenic to mice without prior adaption, whereas JS/08 had limited ability to replicate. Our findings suggest that pigs are naturally infected with avian H5N1 virus and highlight the potential threat to public health due to adaption or reassortment of H5N1 virus in this species.


Videos of conspecifics elicit interactive looking patterns and facial expressions in monkeys.
A broader understanding of the neural basis of social behavior in primates requires the use of species-specific stimuli that elicit spontaneous, but reproducible and tractable behaviors. In this context of natural behaviors, individual variation can further inform about the factors that influence social interactions. To approximate natural social interactions similar to those documented by field studies, we used unedited video footage to induce in viewer monkeys spontaneous facial expressions and looking patterns in the laboratory setting. Three adult male monkeys (Macaca mulatta), previously behaviorally and genetically (5-HTTLPR) characterized, were monitored while they watched 10 s video segments depicting unfamiliar monkeys (movie monkeys) displaying affiliative, neutral, and aggressive behaviors. The gaze and head orientation of the movie monkeys alternated between "averted" and "directed" at the viewer. The viewers were not reinforced for watching the movies, thus their looking patterns indicated their interest and social engagement with the stimuli. The behavior of the movie monkey accounted for differences in the looking patterns and facial expressions displayed by the viewers. We also found multiple significant differences in the behavior of the viewers that correlated with their interest in these stimuli. These socially relevant dynamic stimuli elicited spontaneous social behaviors, such as eye-contact induced reciprocation of facial expression, gaze aversion, and gaze following, that were previously not observed in response to static images. This approach opens a unique opportunity to understanding the mechanisms that trigger spontaneous social behaviors in humans and nonhuman primates.


Human induced pluripotent stem cell-derived MGE cell grafting after status epilepticus attenuates chronic epilepsy and comorbidities via synaptic integration.
Medial ganglionic eminence (MGE)-like interneuron precursors derived from human induced pluripotent stem cells (hiPSCs) are ideal for developing patient-specific cell therapy in temporal lobe epilepsy (TLE). However, their efficacy for alleviating spontaneous recurrent seizures (SRS) or cognitive, memory, and mood impairments has never been tested in models of TLE. Through comprehensive video- electroencephalographic recordings and a battery of behavioral tests in a rat model, we demonstrate that grafting of hiPSC-derived MGE-like interneuron precursors into the hippocampus after status epilepticus (SE) greatly restrained SRS and alleviated cognitive, memory, and mood dysfunction in the chronic phase of TLE. Graft-derived cells survived well, extensively migrated into different subfields of the hippocampus, and differentiated into distinct subclasses of inhibitory interneurons expressing various calcium-binding proteins and neuropeptides. Moreover, grafting of hiPSC-MGE cells after SE mediated several neuroprotective and antiepileptogenic effects in the host hippocampus, as evidenced by reductions in host interneuron loss, abnormal neurogenesis, and aberrant mossy fiber sprouting in the dentate gyrus (DG). Furthermore, axons from graft-derived interneurons made synapses on the dendrites of host excitatory neurons in the DG and the CA1 subfield of the hippocampus, implying an excellent graft-host synaptic integration. Remarkably, seizure-suppressing effects of grafts were significantly reduced when the activity of graft-derived interneurons was silenced by a designer drug while using donor hiPSC-MGE cells expressing designer receptors exclusively activated by designer drugs (DREADDs). These results implied the direct involvement of graft-derived interneurons in seizure control likely through enhanced inhibitory synaptic transmission. Collectively, the results support a patient-specific MGE cell grafting approach for treating TLE.


Effects of spatial smoothing on functional brain networks.
Graph-theoretical methods have rapidly become a standard tool in studies of the structure and function of the human brain. Whereas the structural connectome can be fairly straightforwardly mapped onto a complex network, there are more degrees of freedom in constructing networks that represent functional connections between brain areas. For functional magnetic resonance imaging (fMRI) data, such networks are typically built by aggregating the blood-oxygen-level dependent signal time series of voxels into larger entities (such as Regions of Interest in some brain atlas) and determining their connection strengths from some measure of time-series correlations. Although it is evident that the outcome must be affected by how the voxel-level time series are treated at the preprocessing stage, there is a lack of systematic studies of the effects of preprocessing on network structure. Here, we focus on the effects of spatial smoothing, a standard preprocessing method for fMRI. We apply various levels of spatial smoothing to resting-state fMRI data and measure the changes induced in functional networks. We show that the level of spatial smoothing clearly affects the degrees and other centrality measures of functional network nodes; these changes are non-uniform, systematic, and depend on the geometry of the brain. The composition of the largest connected network component is also affected in a way that artificially increases the similarity of the networks of different subjects. Our conclusion is that wherever possible, spatial smoothing should be avoided when preprocessing fMRI data for network analysis.


Linking Microbial Community Structure and Function During the Acidified Anaerobic Digestion of Grass.
Harvesting valuable bioproducts from various renewable feedstocks is necessary for the critical development of a sustainable bioeconomy. Anaerobic digestion is a well-established technology for the conversion of wastewater and solid feedstocks to energy with the additional potential for production of process intermediates of high market values (e.g., carboxylates). In recent years, first-generation biofuels typically derived from food crops have been widely utilized as a renewable source of energy. The environmental and socioeconomic limitations of such strategy, however, have led to the development of second-generation biofuels utilizing, amongst other feedstocks, lignocellulosic biomass. In this context, the anaerobic digestion of perennial grass holds great promise for the conversion of sustainable renewable feedstock to energy and other process intermediates. The advancement of this technology however, and its implementation for industrial applications, relies on a greater understanding of the microbiome underpinning the process. To this end, microbial communities recovered from replicated anaerobic bioreactors digesting grass were analyzed. The bioreactors leachates were not buffered and acidic pH (between 5.5 and 6.3) prevailed at the time of sampling as a result of microbial activities. Community composition and transcriptionally active taxa were examined using 16S rRNA sequencing and microbial functions were investigated using metaproteomics. Bioreactor fraction, i.e., grass or leachate, was found to be the main discriminator of community analysis across the three molecular level of investigation (DNA, RNA, and proteins). Six taxa, namely Bacteroidia, Betaproteobacteria, Clostridia, Gammaproteobacteria, Methanomicrobia, and Negativicutes accounted for the large majority of the three datasets. The initial stages of grass hydrolysis were carried out by Bacteroidia, Gammaproteobacteria, and Negativicutes in the grass biofilms, in addition to Clostridia in the bioreactor leachates. Numerous glycolytic enzymes and carbohydrate transporters were detected throughout the bioreactors in addition to proteins involved in butanol and lactate production. Finally, evidence of the prevalence of stressful conditions within the bioreactors and particularly impacting Clostridia was observed in the metaproteomes. Taken together, this study highlights the functional importance of Clostridia during the anaerobic digestion of grass and thus research avenues allowing members of this taxon to thrive should be explored.


Statistical Machines for Trauma Hospital Outcomes Research: Application to the PRospective, Observational, Multi-Center Major Trauma Transfusion (PROMMTT) Study.
Improving the treatment of trauma, a leading cause of death worldwide, is of great clinical and public health interest. This analysis introduces flexible statistical methods for estimating center-level effects on individual outcomes in the context of highly variable patient populations, such as those of the PRospective, Observational, Multi-center Major Trauma Transfusion study. Ten US level I trauma centers enrolled a total of 1,245 trauma patients who survived at least 30 minutes after admission and received at least one unit of red blood cells. Outcomes included death, multiple organ failure, substantial bleeding, and transfusion of blood products. The centers involved were classified as either large or small-volume based on the number of massive transfusion patients enrolled during the study period. We focused on estimation of parameters inspired by causal inference, specifically estimated impacts on patient outcomes related to the volume of the trauma hospital that treated them. We defined this association as the change in mean outcomes of interest that would be observed if, contrary to fact, subjects from large-volume sites were treated at small-volume sites (the effect of treatment among the treated). We estimated this parameter using three different methods, some of which use data-adaptive machine learning tools to derive the outcome models, minimizing residual confounding by reducing model misspecification. Differences between unadjusted and adjusted estimators sometimes differed dramatically, demonstrating the need to account for differences in patient characteristics in clinic comparisons. In addition, the estimators based on robust adjustment methods showed potential impacts of hospital volume. For instance, we estimated a survival benefit for patients who were treated at large-volume sites, which was not apparent in simpler, unadjusted comparisons. By removing arbitrary modeling decisions from the estimation process and concentrating on parameters that have more direct policy implications, these potentially automated approaches allow methodological standardization across similar comparativeness effectiveness studies.


De novo demonstration and co-localization of free-radical production and apoptosis formation in rat kidney subjected to ischemia/reperfusion.
Ischemia-induced oxidative damage to the reperfused kidney was examined. A modified chemiluminescence method, an in situ nitro blue tetrazolium perfusion technique, and a DNA fragmentation/apoptosis-related protein assay were adapted for demonstration de novo and co-localization of reactive oxygen species (ROS) production and apoptosis formation in rat kidneys subjected to ischemia/reperfusion injury. The results showed that prolonged ischemia potentiated proapoptotic mechanisms, including increases in the Bax/Bcl-2 ratio, CPP32 expression, and poly-(ADP-ribose)-polymerase fragments, and subsequently resulted in severe apoptosis, including increases in DNA fragmentation and apoptotic cell number in renal proximal tubules (PT) and distal tubules (DT) in a time-dependent manner. The increased level of ROS detected on the renal surface was correlated with that in blood and was intensified by a prolonged interval of ischemia. The main source of ROS synthesis was the PT epithelial cells. The ROS and apoptotic nuclei detected in the PT cells can be ameliorated by superoxide dismutase (SOD) treatment before reperfusion. However, the apoptotic nuclei remained in DT in the SOD-treated rats, indicating that formation of apoptosis in DT was not influenced by the small amounts of ROS produced. In PT and DT cell cultures, significant increases in apoptotic cells and ROS were evident in PT cells after hypoxia/reoxygenation insult. Furthermore, the oxidative damage in PT, but not in DT, can be alleviated by ROS scavengers SOD and hexa(sulfobutyl)fullerene, confirming that PT are vulnerable to ROS. These results lead us to conclude that ROS produced in significant amounts in PT epithelium under ischemia/reperfusion or hypoxia/reoxygenation conditions may be responsible for the apoptotic death of these cells.


Postmortem amitriptyline pharmacokinetics in pigs after oral and intravenous routes of administration.
In this study we have evaluated the postmortem pharmacokinetics of amitriptyline (Ami) and metabolites in pigs after oral and intravenous administration, and the results are compared with previous studies in rats and humans. In addition a meticulous investigation of blood and tissue concentrations after postmortem intravenous infusion of Ami was undertaken. Of a total of 9 over-night fasted pigs, 3 were given 25 mg/Kg Ami orally, and another 3 pigs received an intravenous infusion lasting 1 h of 3.3 mg/Kg Ami prior to death. The final 3 pigs were sacrificed and then given the intravenous infusion after death. After approximately 5 h at room temperature, all carcasses were subsequently stored at 4-5 degrees C. Postmortem blood samples were collected at 0.25, 1, 2, 4, 8, 24, 48, and 96 h through an indwelling intracardial needle. Postmortem examination with blood and tissue sampling was performed 96 h after death. Analysis was carried out by high performance liquid chromatography with ultraviolet detection. Postmortem blood samples from the heart of the orally dosed animals revealed large and variable concentration increases of 99(30-243)% for Ami and 96(52-429)% for the main metabolite 10-OH-Ami at 96 h. In the intravenously infused live pigs heart blood Ami increased by 55(33-69)% and 10-OH-Ami increased by 232(76-240)%. Blood from the atria had significantly higher Ami concentrations than blood from both ventricles in the animals dosed while alive, and the drug concentration in femoral blood was higher than in heart blood (p < 0.01). In the orally dosed pigs the left lobe of the liver had significantly higher Ami levels than the right lobe. Tissue/blood Ami concentration ratios were generally lower than previously reported in rats and approximating the levels reported in humans. The animals infused intravenously after death demonstrated high drug levels in blood samples from central vessels, heart, lungs as well as cerebrospinal fluid and vitreous humour. This implies that the presence of a lethal concentration of a drug in just one sample of heart blood can prove worthless in a case where agonal drug infusion may have occurred.


Initial Laparotomy Versus Peritoneal Drainage in Extremely Low Birthweight Infants With Surgical Necrotizing Enterocolitis or Isolated Intestinal Perforation: A Multicenter Randomized Clinical Trial.
The aim of this study was to determine which initial surgical treatment results in the lowest rate of death or neurodevelopmental impairment (NDI) in premature infants with necrotizing enterocolitis (NEC) or isolated intestinal perforation (IP). The impact of initial laparotomy versus peritoneal drainage for NEC or IP on the rate of death or NDI in extremely low birth weight infants is unknown. We conducted the largest feasible randomized trial in 20 US centers, comparing initial laparotomy versus peritoneal drainage. The primary outcome was a composite of death or NDI at 18 to 22 months corrected age, analyzed using prespecified frequentist and Bayesian approaches. Of 992 eligible infants, 310 were randomized and 96% had primary outcome assessed. Death or NDI occurred in 69% of infants in the laparotomy group versus 70% with drainage [adjusted relative risk (aRR) 1.0; 95% confidence interval (CI): 0.87-1.14]. A preplanned analysis identified an interaction between preoperative diagnosis and treatment group (P = 0.03). With a preoperative diagnosis of NEC, death or NDI occurred in 69% after laparotomy versus 85% with drainage (aRR 0.81; 95% CI: 0.64-1.04). The Bayesian posterior probability that laparotomy was beneficial (risk difference <0) for a preoperative diagnosis of NEC was 97%. For preoperative diagnosis of IP, death or NDI occurred in 69% after laparotomy versus 63% with drainage (aRR, 1.11; 95% CI: 0.95-1.31); Bayesian probability of benefit with laparotomy = 18%. There was no overall difference in death or NDI rates at 18 to 22 months corrected age between initial laparotomy versus drainage. However, the preoperative diagnosis of NEC or IP modified the impact of initial treatment.


A survey of the mycorrhization of Southeast Asian ferns and lycophytes.
The colonisation of land by plants may not have been possible without mycorrhizae, which supply the majority of land plants with nutrients, water and other benefits. In this sense, the mycorrhization of basal groups of land plants such as ferns and lycophytes is of particular interest, yet only about 9% of fern and lycophyte species have been sampled for their mycorrhization status, and no community-level analyses exist for tropical fern communities. In the present study, we screened 170 specimens of ferns and lycophytes from Malaysia and Sulawesi (Indonesia), representing 126 species, and report the mycorrhization status for 109 species and 19 genera for the first time. Mycorrhizal colonisations were detected in 96 (56.5%) of the specimens, 85 of which corresponded to arbuscular mycorrhizae (AMF), three to dark-septate endophytes (DSE) and four to mixed colonisations (AMF + DSE). DSE colonisations were lower than in comparable samples of ferns from the Andes, suggesting a geographical or taxonomic pattern in this type of colonisation. Epiphytes had significantly lower levels of colonisation (26.1%) than terrestrial plants (70.7%), probably due to the difficulty of establishment of mycorrhizal fungi in the canopy habitat.


In vivo magnetic resonance imaging and spectroscopy identifies oncolytic adenovirus responders.
At present, it is not possible to reliably identify patients who will benefit from oncolytic virus treatments. Conventional modalities such as computed tomography (CT), which measure tumor size, are unreliable owing to inflammation-induced tumor swelling. We hypothesized that magnetic resonance imaging (MRI) and spectroscopy (MRS) might be useful in this regard. However, little previous data exist and neither oncolytic adenovirus nor immunocompetent models have been assessed by MRS. Here, we provide evidence that in T2-weighted MRI a hypointense core area, consistent with coagulative necrosis, develops in immunocompetent Syrian hamster carcinomas that respond to oncolytic adenovirus treatment. The same phenomenon was observed in a neuroblastoma patient while he responded to the treatment. With relapse at a later stage, however, the tumor of this patient became moderately hyperintense. We found that MRS of taurine, choline and unsaturated fatty acids can be useful early indicators of response and provide detailed information about tumor growth and degeneration. In hamsters, calprotectin-positive inflammatory cells (heterophils and macrophages) were found in abundance; particularly surrounding necrotic areas in carcinomas and T cells were significantly increased in sarcomas, when these had been treated with a granulocyte-macrophage colony-stimulating factor-producing virus, suggesting a possible link between oncolysis, necrosis (seen as a hypointense core in MRI) and/or immune response. Our study indicates that both MRI and MRS could be useful in the estimation of oncolytic adenovirus efficacy at early time points after treatment.


Morphological Control of Coil-Rod-Coil Molecules Containing m-Terphenyl Group: Construction of Helical Fibers and Helical Nanorings in Aqueous Solution.
Rod-coil molecules, composed of rigid segments and flexible coil chains, have a strong intrinsic ability to self-assemble into diverse supramolecular nanostructures. Herein, we report the synthesis and the morphological control of a new series of amphiphilic coil-rod-coil molecular isomers 1-2 containing flexible oligoether chains. These molecules are comprised of m-terphenyl and biphenyl groups, along with triple bonds, and possess lateral methyl or butyl groups at the coil or rod segments. The results of this study suggest that the morphology of supramolecular aggregates is significantly influenced by the lateral alkyl groups and by the sequence of the rigid fragments in the bulk and in aqueous solution. The molecules with different coils self-assemble into lamellar or oblique columnar structures in the bulk state. In aqueous solution, molecule 1a, with a lack of lateral groups, self-assembled into large strips of sheets, whereas exquisite nanostructures of helical fibers were obtained from molecule 1b, which incorporated lateral methyl groups between the rod and coil segments. Interestingly, molecule 1c with lateral butyl and methyl groups exhibited a strong self-organizing capacity to form helical nanorings. Nanoribbons, helical fibers, and small nanorings were simultaneously formed from the 2a-2c, which are structural isomers of 1a, 1b, and 1c. Accurate control of these supramolecular nanostructures can be achieved by tuning the synergistic interactions of the noncovalent driving force with hydrophilic-hydrophobic interactions in aqueous solution.


Effects of radiofrequency electromagnetic wave exposure from cellular phones on the reproductive pattern in male Wistar rats.
The present study investigates the effect of free radical formation due to mobile phone exposure and effect on fertility pattern in 70-day-old male Wistar rats (sham exposed and exposed). Exposure took place in Plexiglas cages for 2 h a day for 35 days to mobile phone frequency. The specific absorption rate was estimated to be 0.9 W/kg. An analysis of antioxidant enzymes glutathione peroxidase (P < 0.001) and superoxide dismutase (P < 0.007) showed a decrease, while an increase in catalase (P < 0.005) was observed. Malondialdehyde (P < 0.003) showed an increase and histone kinase (P = 0.006) showed a significant decrease in the exposed group. Micronuclei also show a significant decrease (P < 0.002) in the exposed group. A significant change in sperm cell cycle of G(0)-G(1) (P = 0.042) and G(2)/M (P = 0.022) were recorded. Generation of free radicals was recorded to be significantly increased (P = 0.035). Our findings on antioxidant, malondialdehyde, histone kinase, micronuclei, and sperm cell cycle are clear indications of an infertility pattern, initiated due to an overproduction of reactive oxygen species. It is concluded that radiofrequency electromagnetic wave from commercially available cell phones might affect the fertilizing potential of spermatozoa.


[Adverse drug reactions in patients attending in emergency service].
Describing adverse drug reactions (ADR) and analyzing the factors associated with their appearance in patients attending two hospitals' emergency services. Descriptive, medicine-surveillance study of people consulting the emergency service at the Rosales hospital and the basic total health attention unit in Pereira, Colombia, between July and September 2005 for any symptom related to taking medicine. Naranjo's Adverse Drug Reaction probability scale was used. A total of 91 patients were found; 62,6 % were women having an average age of 36,3+/-22,4 (range 0-85). Antibiotics (24,2 %) NSAIDs (17,6 %), analgesics (9,9 %) and antidiabetic agents (8,8 %) were the most used drugs. 39,6 % of the patients only used one medicine; the average of drugs per patient was 2,4+/-1,5. Self-medication was present in 25,3 %. The most common ADRs arose from urticaria (31,9 %), hypoglycaemia (8,8 %), gastritis (6,6 %), angioedema (5,5 %) and anaphylaxis (4,4 %). 13,2 % of ADRs were serious and 54,9 % were avoidable. 14,3 % of ADRs were classified as being definite and 75,8 % probable. Only 14,3 % of patients had a past history of ADR. The average cost of being attended was US$ 78,1 dollars. Analgesic use was associated with anaphylaxis, antibiotics with urticaria whilst NSAIDs was associated with gastritis and gastrointestinal bleeding, antidiabetic agents with hypoglycaemia, antipsychotic and antidepressants agents with neurological symptoms and warfarin with bleeding. The most important ADR-associated risk factors in patients attending emergency services were determined. Special care must be paid to patients using NSAIDs, warfarin, metoclopramide, metamizole, antipsychotic and antidepressants agents, antidiabetic agents and antibiotics, having a past history of ADR, being more than 55 years old and presenting urticaria, digestive and/or neurological symptoms


Management of immune-related adverse events and kinetics of response with ipilimumab.
Monoclonal antibodies directed against the immune checkpoint protein cytotoxic T-lymphocyte antigen-4 (CTLA-4; CD152)-ipilimumab and tremelimumab-have been investigated in metastatic melanoma and other cancers and have shown promising results. Recently, ipilimumab was approved by the US Food and Drug Administration for the treatment of metastatic melanoma. We review the literature on managing the adverse effects and kinetics of tumor regression with ipilimumab and provide guidelines on their management. During treatment with these antibodies, a unique set of adverse effects may occur, called immune-related adverse events (irAEs). These include rashes, which may rarely progress to life-threatening toxic epidermal necrolysis, and colitis, characterized by a mild to moderate, but occasionally also severe and persistent diarrhea. Hypophysitis, hepatitis, pancreatitis, iridocyclitis, lymphadenopathy, neuropathies, and nephritis have also been reported with ipilimumab. Early recognition of irAEs and initiation of treatment are critical to reduce the risk of sequelae. Interestingly, irAEs correlated with treatment response in some studies. Unique kinetics of response have been observed with CTLA-4 blockade with at least four patterns: (1) response in baseline lesions by week 12, with no new lesions seen; (2) stable disease, followed by a slow, steady decline in total tumor burden; (3) regression of tumor after initial increase in total tumor burden; and (4) reduction in total tumor burden during or after the appearance of new lesion(s) after week 12. We provide a detailed description of irAEs and recommendations for practicing oncologists who are managing them, along with the unusual kinetics of response associated with ipilimumab therapy.


Molecular Profiling of Patients with Pancreatic Cancer: Initial Results from the Know Your Tumor Initiative.
Purpose: To broaden access to and implementation of precision medicine in the care of patients with pancreatic cancer, the Know Your Tumor (KYT) program was initiated using a turn-key precision medicine system. Patients undergo commercially available multiomic profiling to determine molecularly rationalized clinical trials and off-label therapies.Experimental Design: Tumor samples were obtained for 640 patients from 287 academic and community practices covering 44 states. College of American Pathologists/Clinical Laboratory Improvement Amendments-accredited laboratories were used for genomic, proteomic, and phosphoprotein-based molecular profiling.Results: Tumor samples were adequate for next-generation sequencing in 96% and IHC in 91% of patients. A tumor board reviewed the results for every patient and found actionable genomic alterations in 50% of patients (with 27% highly actionable) and actionable proteomic alterations (excluding chemopredictive markers) in 5%. Actionable alterations commonly found were in DNA repair genes (BRCA1/2 or ATM mutations, 8.4%) and cell-cycle genes (CCND1/2/3 or CDK4/6 alterations, 8.1%). A subset of samples was assessed for actionable phosphoprotein markers. Among patients with highly actionable biomarkers, those who received matched therapy (n = 17) had a significantly longer median progression-free survival (PFS) than those who received unmatched therapy [n = 18; PFS = 4.1 vs. 1.9 months; HR, 0.47; 95% confidence interval (CI): 0.24-0.94; P adj = 0.03].Conclusions: A comprehensive precision medicine system can be implemented in community and academic settings, with highly actionable findings observed in over 25% of pancreatic cancers. Patients whose tumors have highly actionable alterations and receive matched therapy demonstrated significantly increased PFS. Our findings support further prospective evaluation of precision oncology in pancreatic cancer. Clin Cancer Res; 24(20); 5018-27. ©2018 AACR.


Methodological and theoretical improvements in the study of superstitious beliefs and behaviour.
Via four studies (N = 901), we developed an improved Belief in Superstition Scale (BSS) composed of three distinct components (belief in bad luck, belief in good luck, and the belief that luck can be changed), whose structure was supported through exploratory (Study 1) and confirmatory (Studies 2 and 3) factor analyses using divergent samples. We found that among theoretical predictors, higher 'chance' locus of control (i.e., the belief that chance/fate controls one's life) best predicted all three BSS subscales (Studies 2-3). In Study 3, we found that BSS subscale scores were reliable, but largely invariant across age and education with a non-general psychology sample. In Study 4, the BSS subscales best predicted participants' superstitious attitudes and behaviour in a new lottery drawing paradigm among other commonly used superstition scales. Taken together, our results indicate that the BSS is a valuable addition to the burgeoning research on superstitious attitudes and behaviour.


Magnetic resonance imaging in the evaluation of (deep infiltrating) endometriosis: the value of diffusion-weighted imaging.
To assess the value of magnetic resonance (MR)diffusion-weighted imaging (DWI) in the evaluation of deep infiltrating endometriosis (DIE). In a prospective single-center study, DWI was added to the standard MRI protocol in 56 consecutive patients with known or suspected endometriosis. Endometriotic lesions as well as (functional) ovarian cysts were analyzed for location, size, and signal intensity on T1, T2, and DWI. Apparent diffusion coefficient (ADC) values were calculated using b-values of 50, 400, 800,and 1200 s/mm(2). Statistical analysis included the Spearman correlation coefficient, Mann-Whitney U, and Kruskal-Wallis tests. A total of 110 lesions (62 endometrial cysts and 48 DIE) were detected, 60 of which were large enough to analyze. Mean ADC values of endometrial cysts and functional ovarian cysts were 1.10 x 10(-3)/mm(2)/s and 2.14 x 10(-3)/mm(2)/s, respectively. Mean ADC values of DIE retrocervical, infiltrating the colon, and bladder were 0.70 x 10(-3)/mm(2)/s, 0.77 x 10(-3)/mm(2)/s, and 0.79 x 10(-3)/mm(2)/s, respectively. ADC values of DIE did not show a significant difference between varying pelvic locations (P = 0.63). Results of our study suggest that ADC values of DIE are consistently low, without significant difference between pelvic locations.


Noninvasive positive pressure ventilation enhances the effects of aerobic training on cardiopulmonary function.
The purpose of this study was to determine the effect of aerobic training under noninvasive positive pressure ventilation (NPPV) on maximal oxygen uptake ([Formula: see text]). Ten healthy young male volunteers participated in the study. Before the training, stroke volume (SV) and cardiac output (CO) were measured in all subjects under 0, 4, 8, and 12 cmH2O NPPV at rest. Then, the subjects exercised on a cycle ergometer at 60% of pre-training [Formula: see text] for 30 min daily for 5 consecutive days with/without NPPV. The 5-day exercise protocol was repeated after a three-week washout period without/with NPPV. The primary endpoint was changes in [Formula: see text]. The secondary endpoints were changes in SV, CO, maximum heart rate (HRmax), maximum respiratory rate (RRmax), maximum expiratory minute volume (VEmax) and the percent change in plasma volume (PV). NPPV at 12 cmH2O significantly reduced SV and CO at rest. [Formula: see text] significantly increased after 5 days training with and without NPPV, but the magnitude of increase in [Formula: see text] after training under 12 cmH2O NPPV was significantly higher than after training without NPPV. VEmax significantly increased after training under NPPV, but not after training without NPPV. HRmax and RRmax did not change during training irrespective of NPPV. The percent change in PV was similar between training with and without NPPV. The 5-day training program with NPPV resulted in greater improvement in [Formula: see text] than without NPPV. Aerobic training under NPPV has add-on effects on [Formula: see text] and exercise-related health benefits in healthy young men.


Diets of drinkers on drinking and nondrinking days: NHANES 2003-2008.
Alcohol may affect dietary intake. However, little is known about diets on drinking days in the US population. We determined whether the diets of drinkers differ on drinking compared with nondrinking days. Data were from the 2003-2008 NHANES Mobile Examination Center interview. We identified 1864 current drinkers (1126 men and 738 women) who completed two 24-h dietary recalls, one of which was on a drinking day and the other of which was on a nondrinking day. Sex-specific repeated-measures analyses that were adjusted for dietary recall order and recall day of the week were used to compare within-individual differences in energy, nutrient, and food-group intakes. Analyses were weighted to produce representative estimates. On their drinking (compared with nondrinking) days, men consumed an excess 168 nonalcohol kcal (P < 0.01), which was reflected in higher intakes of nutrients including total protein (P < 0.001), total fat (P < 0.01), saturated fat (P < 0.01), monounsaturated fat (P < 0.01), potassium (P < 0.001), and sodium (P < 0.05). Men also had higher intakes of food groups including meat (P < 0.001), white potatoes (P < 0.05), and discretionary oil and solid fat (P < 0.05) and lower intakes of total fruit (P < 0.05) and milk (P < 0.05). Women did not consume excess nonalcohol kilocalories but had higher intakes of total fat (P < 0.05), monounsaturated fat (P < 0.05), polyunsaturated fat (P < 0.05), potassium (P < 0.01), and discretionary oil and solid fat (P < 0.05) and lower intakes of milk (P < 0.01) and milk products (P < 0.01). These mostly moderate drinkers had poorer diets on drinking days. Same-day associations between alcohol and diet could be useful targets for public health efforts to improve dietary intake.


Controlled release of drugs from multi-component biomaterials.
In order to control their release, drugs are encapsulated into systems which are expected to provide a certain site with a predetermined amount of drug over a well-defined period of time. Here we report on a multi-component drug delivery biomaterial that consists of a hydrogel matrix in which drug-loaded biodegradable microcarriers are dispersed, and whose potential applications could be found in the design of implantable devices with long-term activity, as required by contraceptive and hormone replacement treatments. The release profile of the drug can actually be tuned by the complex interplay of several release mechanisms, including the permeability and eventually the degradation rate of the microcarriers and the diffusion through the hydrogel. The hydrogel consisted of 2-hydroxyethyl methacrylate cross-linked by ethylene glycol dimethacrylate. The microcarriers were biodegradable poly-epsilon-caprolactone (PCL) microspheres in which active molecules, such as levonorgestrel (LNG), were encapsulated. The hydrogels were characterized by water swelling, thermal properties, LNG diffusion through drug-free and drug-depleted hydrogel membranes and LNG release from devices with drug dispersed in the hydrogel. The PCL microspheres were observed by scanning electron microscopy; their size distribution, LNG loading and release were also investigated. The hydrogel-microsphere assemblies were characterized in terms of the distribution of the microspheres within the hydrogel, water swelling and the release of the encapsulated molecules. The developed device, due to its composite structure, has the ability to combine several release mechanisms, leading to drug release obeying zero-order kinetics for most of the time.


Proline and hydroxyproline excretion and vitamin C status in elderly human subjects.
1. Plasma and buffy-coat vitamin C, urinary proline, hydroxyproline, creatinine and total amino acid concentrations were meausred in 23 healthy elderly subjects at intervals of 3 months. 2. There was a strong positive correlation between plasma vitamin C and buffy-coat vititamin C. 3. There were not significant correlations between plasma or buffy-coat vitamin CPAMIN C. 3. There were not significant correlations between plasma or buffy-coat vitamin C and total urinary hydroxyproline, whether expressed on a creatinine basis or on a total amino acid basis. Similarly, no significant correlations could be detected involving the proline/hydroxyproline ratio in urine hydrolysates. 4. There was a significant negative correlation between plasma or buffy-coat vitamin C and total urinary proline, when expressed per unit of total urinary proline, when expressed per unit of total urinary proline, when expressed per unit of total amino acids in the hydrolysates. This correlation was not observed with unhydrolysed urine, and it appeared to reside in the diffusible fraction, part of whose proline could be liberated by prolidase digestion. In addition, in the man, there was some evidence for a positive correlation between plasma or buffy-coat vitamin C and the ratio of total urinary amino acids to creatinine. 5. These results support the view that poor vitamin C status in elderly humans may be associated with a defect in collagen proline hydroxylation, reflected by increased excretion of proline-rich, collagen-derived peptides. If this interpretation is correct, it indicates a potential defect in connective tissue repair, and could form the basis of a functional test for subclinical vitamin C deficiency.


The mechanism of the phosphoramidite synthesis of polynucleotides.
The mechanism of the coupling step in polynucleotide synthesis using 5'-4,4'-dimethoxytritylthymidine-3'-beta-cyanoethyl-N,N-diisopropylphosphoramidite as the phosphitylating agent and catalysed by the salt of saccharin and N-methylimidazole in acetonitrile has been studied by (31)P NMR. Pre- and post-equilibria between the activator salt and released diisopropylamine have been examined by (1)H NMR and ITC, which show that the salt between saccharin and diisopropylamine will be present in acetonitrile. Activation of the phosphoramidite by the salt of saccharin and N-methylimidazole involves nucleophilic catalysis and the formation of a reactive saccharin adduct bonded through its carbonyl oxygen to phosphorus. The rate constants for the reaction of the 4-methoxyphenol with 5'-4,4'-dimethoxytritylthymidine-3'-beta-cyanoethyl-N,N-diisopropylphosphoramidite in the presence of saccharin-N-methylimidazole salt show a non-linear dependence on phenol concentration, becoming independent at high phenol concentrations, compatible with a change in rate limiting step from the alcoholysis step to the activation step.


Mechanism of repeat-associated microRNAs in fragile X syndrome.
The majority of the human genome is comprised of non-coding DNA, which frequently contains redundant microsatellite-like trinucleotide repeats. Many of these trinucleotide repeats are involved in triplet repeat expansion diseases (TREDs) such as fragile X syndrome (FXS). After transcription, the trinucleotide repeats can fold into RNA hairpins and are further processed by Dicer endoribonuclases to form microRNA (miRNA)-like molecules that are capable of triggering targeted gene-silencing effects in the TREDs. However, the function of these repeat-associated miRNAs (ramRNAs) is unclear. To solve this question, we identified the first native ramRNA in FXS and successfully developed a transgenic zebrafish model for studying its function. Our studies showed that ramRNA-induced DNA methylation of the FMR1 5'-UTR CGG trinucleotide repeat expansion is responsible for both pathological and neurocognitive characteristics linked to the transcriptional FMR1 gene inactivation and the deficiency of its protein product FMRP. FMRP deficiency often causes synapse deformity in the neurons essential for cognition and memory activities, while FMR1 inactivation augments metabotropic glutamate receptor (mGluR)-activated long-term depression (LTD), leading to abnormal neuronal responses in FXS. Using this novel animal model, we may further dissect the etiological mechanisms of TREDs, with the hope of providing insights into new means for therapeutic intervention.


The prognostic effect of brain natriuretic peptide levels on outcomes of hospitalized patients with COVID-19.
Natriuretic peptides are biomarkers of myocardial stress and are frequently elevated among patients with severe respiratory illnesses, typically in the absence of elevated cardiac-filling pressures or clinical heart failure. Elevation of brain natriuretic peptide (BNP) or NT-proBNP is associated with worse outcomes among patients with Acute Respiratory Distress Syndrome (ARDS). We conducted a retrospective cohort study based on a comprehensive review of Electronic Medical Records (EMRs) of patients with Coronavirus Disease 2019 (COVID-19) to evaluate whether BNP on admission has prognostic value on mortality and hospital length of stay (LOS) among patients admitted with confirmed COVID-19 along with the inclusion of additional prognostic variables. Overall, 146 patients were included after analyzing 230 patients' EMR and excluding potential confounding factors for abnormal BNP. Our statistical analysis did not show a statistically significant association between BNP level and mortality rate (P = 0.722) or ICU LOS ( P = 0.741). A remarkable secondary outcome to our study was that impaired renal function (GFR<60) on admission was significantly associated with an increased mortality rate (P = 0.026) and an increased ICU LOS (P = 0.022). Although various studies have presented the predictive role of pro-BNP among patients with respiratory distress in the past years, our study did not find BNP to be an accurate predictive and prognostic factor among patients with COVID-19 in our study population. Renal impairment and high Acute Physiology and Chronic Health Evaluation (APACHE) II scores on admission, on the other hand, have demonstrated to be strong predictors for COVID-19 morbidity and mortality. This study could represent an introduction to more prominent multicenter studies to evaluate additional prognostic factors and minimize the ordering of nonspecific testing.


Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli.
Sulforaphane, an isothiocyanate from broccoli, is one of the most potent food-derived anticarcinogens. This compound is not present in the intact vegetable, rather it is formed from its glucosinolate precursor, glucoraphanin, by the action of myrosinase, a thioglucosidase enzyme, when broccoli tissue is crushed or chewed. However, a number of studies have demonstrated that sulforaphane yield from glucoraphanin is low, and that a non-bioactive nitrile analog, sulforaphane nitrile, is the primary hydrolysis product when plant tissue is crushed at room temperature. Recent evidence suggests that in Arabidopsis, nitrile formation from glucosinolates is controlled by a heat-sensitive protein, epithiospecifier protein (ESP), a non-catalytic cofactor of myrosinase. Our objectives were to examine the effects of heating broccoli florets and sprouts on sulforaphane and sulforaphane nitrile formation, to determine if broccoli contains ESP activity, then to correlate heat-dependent changes in ESP activity, sulforaphane content and bioactivity, as measured by induction of the phase II detoxification enzyme quinone reductase (QR) in cell culture. Heating fresh broccoli florets or broccoli sprouts to 60 degrees C prior to homogenization simultaneously increased sulforaphane formation and decreased sulforaphane nitrile formation. A significant loss of ESP activity paralleled the decrease in sulforaphane nitrile formation. Heating to 70 degrees C and above decreased the formation of both products in broccoli florets, but not in broccoli sprouts. The induction of QR in cultured mouse hepatoma Hepa lclc7 cells paralleled increases in sulforaphane formation.


Endemic HBV among hospital in-patients in Bangladesh, including evidence of occult infection.
Bangladesh is one of the top-ten most heavily burdened countries for viral hepatitis, with hepatitis B (HBV) infections responsible for the majority of cases. Recombinant and occult HBV infections (OBI) have been reported previously in the region. We investigated an adult fever cohort (n=201) recruited in Dhaka, to determine the prevalence of HBV and OBI. A target-enrichment deep sequencing pipeline was applied to samples with HBV DNA >3.0 log10 IU ml-1. HBV infection was present in 16/201 (8 %), among whom 3/16 (19 %) were defined as OBI (HBsAg-negative but detectable HBV DNA). Whole genome deep sequences (WGS) were obtained for four cases, identifying genotypes A, C and D. One OBI case had sufficient DNA for sequencing, revealing multiple polymorphisms in the surface gene that may contribute to the occult phenotype. We identified mutations associated with nucleos(t)ide analogue resistance in 3/4 samples sequenced, although the clinical significance in this cohort is unknown. The high prevalence of HBV in this setting illustrates the importance of opportunistic clinical screening and DNA testing of transfusion products to minimise OBI transmission. WGS can inform understanding of diverse disease phenotypes, supporting progress towards international targets for HBV elimination.


Psychophysiologic responses of mechanically ventilated patients to music: a pilot study.
Although mechanically ventilated patients experience numerous stressors, they have not been included in music therapy stress reduction and relaxation studies. To examine selected psychophysiologic responses of mechanically ventilated patients to music. A two-group experimental design with pretest, posttest, and repeated measures was used. Twenty mechanically ventilated patients were randomized to a music-listening group or a nonmusic (headphones only) group. Physiologic dependent measures--heart rate and rhythm, respiratory rate, systolic and diastolic blood pressure, oxygen saturation, and airway pressure--were collected at timed intervals. Psychologic data were collected before and after intervention using the Profile of Mood States. Using repeated measures analysis of variance, results for heart rate and respiratory rate over time and over time between groups were significant. Between-group differences were significant for respiratory rate. Significant differences were found via t test for the music group's Profile of Mood States scores. No adverse cardiovascular responses were noted for either group. Data indicated that music listening decreased heart rate, respiratory rate, and Profile of Mood States scores, indicating relaxation and mood improvement.


Divided and disconnected--an examination of youths' experiences with emotional distress within the context of their everyday lives.
This paper is based on a qualitative study conducted in a rural community in British Columbia, Canada. Ethnographic methods were used to: (1) to bring youth voice to the literature on emotional distress; and (2) to capture the ways in which context shapes young peoples' experiences of emotional distress within their everyday lives. Our findings demonstrate how socio-structural contextual factors such as the local economy, geographical segregation, racism, ageism, and cutbacks in health and social service programming operate to create various forms of disconnection, and intersect in young peoples' lives to shape their experiences of emotional distress.


Fragments of the internal transcribed spacer 1 of pre-rRNA accumulate in Saccharomyces cerevisiae lacking 5'----3' exoribonuclease 1.
The portion of the internal transcribed spacer 1 found on 20S pre-rRNA accumulates in Saccharomyces cerevisiae lacking 5'----3' exoribonuclease 1, showing that an endonucleolytic cleavage at the 3' terminus of 18S rRNA is involved in the 20S pre-rRNA to 18S mature rRNA conversion. Smaller fragments of the spacer sequence are also found. The exoribonuclease may be involved as a cytoplasmic RNase in the hydrolysis of the spacer.


The Lipocalin LPR-1 Cooperates with LIN-3/EGF Signaling To Maintain Narrow Tube Integrity in Caenorhabditis elegans.
Lipocalins are secreted cup-shaped glycoproteins that bind sterols, fatty acids, and other lipophilic molecules. Lipocalins have been implicated in a wide array of processes related to lipophilic cargo transport, sequestration, and signaling, and several are used as biomarkers for human disease, but the functions of most lipocalins remain poorly understood. Here we show that the Caenorhabditis elegans lipocalin LPR-1 is required to maintain apical membrane integrity and a continuous lumen in two narrow unicellular tubes, the excretory duct and pore, during a period of rapid lumen elongation. LPR-1 fusion protein is expressed by the duct and pore and accumulates both intracellularly and in apical extracellular compartments, but it can also function cell nonautonomously when provided from outside of the excretory system. lpr-1 mutant defects can be rescued by increased signaling through the epidermal growth factor (EGF)-Ras-extracellular signal regulated kinase (ERK) pathway, which promotes the more elongated duct vs. less elongated pore tube fate. Spatial and temporal rescue experiments indicate that Ras signaling acts within the duct and pore tubes during or prior to cell fate determination to bypass the requirement for LPR-1 lpr-1 mutations did not disrupt LIN-3/EGF-dependent duct-fate specification, prevent functioning of any specific LIN-3/EGF isoform, or alter LET-23/EGFR localization, and reduced signaling did not phenocopy or enhance lpr-1 mutant defects. These data suggest that LPR-1 protects lumen integrity through a LIN-3/EGF-independent mechanism, but that increased signaling upregulates some target(s) that can compensate for lpr-1 absence.


Relationships between milk characteristics and somatic cell score in milk from primiparous browsing goats.
To determine milk yield and composition, total microbic count (TMC) and somatic cell count (SCC) of browsing goats throughout the first lactation, 100 goats of Sarda breed, equally distributed in four flocks (F1, F2, F3 and F4), were selected. They were exclusively fed pasture and hand-milked once daily. Individual milk samples and daily milk yield were taken from each goat at monthly intervals, from March to July. Milk samples were analyzed for: total protein, fat, lactose, urea, freezing point (FP), pH, TMC and SCC. The data was subjected to analysis of variance and to correlation matrix. On the whole, in all the flocks, milk yield showed the highest production in April and May. Fat content increased (P < 0.01) throughout the lactation. Protein content showed the lowest value (P < 0.01) in June (4.15%). Urea and pH values were fluctuating. FP was lower (P < 0.01) at the start of lactation (-0.562 Hortvet degrees). TMC log10 values were low, considering the hand milking and inadequacy of facilities on the farms. SCC increased (P < 0.01) throughout the lactation and, on the whole, SCC and TMC were not correlated.


Towards the integration of foreign DNA into the chromosome of Mycobacterium leprae.
Integrative plasmid vectors based on the pSAM2 system of Streptomyces ambofaciens offer great potential for the genetic analysis of Mycobacterium leprae. To assess this, the chromosomal attachment site of M. leprae, att-pSAM2, has been cloned, mapped and characterized. Nucleotide sequence analysis shows att-pSAM2 to correspond to a putative tRNA(pro) gene identical in sequence to those of S. ambofaciens and M. tuberculosis. In addition, it is shown that the genes encoding aspartate semialdehyde dehydrogenase, asd, and an anonymous protein antigen recognized by sera from leprosy patients, are linked to the M. leprae att-pSAM2 locus.


Assessment of quality of care for the management of type 2 diabetes: a multicentre study from a developing country.
Complications of diabetes can be prevented or delayed by providing high quality care. This study aimed to document the quality of care provided to people with type 2 diabetes in Pakistan and to identify the difference in care offered at various clinics. Cross-sectional multi-centre study. Interviews were undertaken with 672 people with type 2 diabetes attending three different types of diabetes clinic (private clinic (A), non-governmental organisation (B) and public clinic (C)) in Karachi, Pakistan. A structured questionnaire was used to collect socio-demographic and clinical information from patients; quality of care indicators were also confirmed by reference to patients' medical records. Overall, 68% (A: 92%, B: 58% and C: 52%, P<0.001) of study subjects were informed about diabetes complications. Blood pressure (BP) monitoring at every visit was completed for 80% of study respondents (A: 100%, B: 79% and C: 57%, P<0.001). Foot examination was infrequent (53%, A: 98%, B: 52% and C: 8% (P<0.001). Lipid profiles of 48% of patients had been done in the past 12 months (A: 77%, B: 16% and C: 50%, P<0.001). Microalbumin testing had been performed in 32% of patients in the previous year (A: 77%, B: 09% and C: 05%; P<0.001). Most participants had elevated glycaemic (58.2%) and BP levels (84.7%) with higher prevalence among people who attended clinics B and C (P<0.001). Overall, 82.6% of study subjects had an elevated body mass index; this was almost equally prevalent across clinics. Many patients with type 2 diabetes do not receive optimal diabetes care in Karachi. Among the different settings, care provided in private health sector clinics was of a better standard. However, our results reveal a need for overall improvement in the quality of diabetes care. Further research is also needed to evaluate the reasons for poor diabetes care, and to identify the most cost-effective means to address these.


Disturbed habitats locally reduce the signal of deep evolutionary history in functional traits of plants.
The functioning of present ecosystems reflects deep evolutionary history of locally cooccurring species if their functional traits show high phylogenetic signal (PS). However, we do not understand what drives local PS. We hypothesize that local PS is high in undisturbed and stressful habitats, either due to ongoing local assembly of species that maintained ancestral traits, or to past evolutionary maintenance of ancestral traits within habitat species-pools, or to both. We quantified PS and diversity of 10 traits within 6704 local plant communities across 38 Dutch habitat types differing in disturbance or stress. Mean local PS varied 50-fold among habitat types, often independently of phylogenetic or trait diversity. Mean local PS decreased with disturbance but showed no consistent relationship to stress. Mean local PS exceeded species-pool PS, reflecting nonrandom subsampling from the pool. Disturbance or stress related more strongly to mean local than to species-pool PS. Disturbed habitats harbour species with evolutionary divergent trait values, probably driven by ongoing, local assembly of species: environmental fluctuations might maintain different trait values within lineages through an evolutionary storage effect. If functional traits do not reflect phylogeny, ecosystem functioning might not be contingent on the presence of particular lineages, and lineages might establish evolutionarily novel interactions.


Proteomic Analysis of the Bovine and Human Ciliary Zonule.
The zonule of Zinn (ciliary zonule) is a system of fibers that centers the crystalline lens on the optical axis of the eye. Mutations in zonule components underlie syndromic conditions associated with a broad range of ocular pathologies, including microspherophakia and ectopia lentis. Here, we used HPLC-mass spectrometry to determine the molecular composition of the zonule. Tryptic digests of human and bovine zonular samples were analyzed by HPLC-mass spectrometry. The distribution of selected components was confirmed by immunofluorescence confocal microscopy. In bovine samples, the composition of the equatorial zonule was compared to that of the hyaloid zonule and vitreous humor. The 52 proteins common to the zonules of both species accounted for >95% of the zonular protein. Glycoproteins constituted the main structural components, with two proteins, FBN1 and LTBP2, constituting 70%-80% of the protein. Other abundant components were MFAP2, EMILIN-1, and ADAMTSL-6. Lysyl oxidase-like 1, a crosslinking enzyme implicated in collagen and elastin biogenesis, was detected at significant levels. The equatorial and hyaloid zonular samples were compositionally similar to each other, although the hyaloid sample was relatively enriched in the proteoglycan opticin and the fibrillar collagens COL2A1, COL11A1, COL5A2, and COL5A3. The zonular proteome was surprisingly complex. In addition to structural components, it contained signaling proteins, protease inhibitors, and crosslinking enzymes. The equatorial and hyaloid zonules were similar in composition, but the latter may form part of a composite structure, the hyaloid membrane, that stabilizes the vitreous face.


Communicable Diseases: Achievements and Challenges for Public Health.
The past two centuries have seen enormous achievements in control of infectious diseases, previously the leading cause of death, in large measure due to sanitation and food safety, vaccines, antibiotics and improved nutrition. This has led people to put their faith in the notion that medical science would succeed in overcoming the remaining obstacles. Vaccination has eradicated smallpox, nearly eradicated poliomyelitis and greatly reduced many other highly dangerous infections such as diphtheria, tetanus and measles. New diseases such as HIV and new forms of influenza have taken both professional and popular opinion by surprise and have renewed the challenges before the world public health community. Emergence of antibiotic-resistant strains of common organisms due to overuse of antibiotics and lack of vaccines for many dangerous microorganisms poses problems to humanity. This stresses the need for new vaccines, effective antibiotics and strengthened environmental control measures. New knowledge of the microbiological origins of cancers such as that of the cervix, stomach and liver have strengthened primary prevention and brought hope that new cures will be found for other chronic diseases of infectious origin. Tragically long delays in adopting "new" and cost effective vaccines cause hundreds of thousands of preventable deaths each year in developing and mid-level developed countries. Gains are being made in control of many tropical diseases, but malaria, tuberculosis and other infectious diseases remain enormous global problems. Research and acquisition of new knowledge, risk communication, application of currently available means and fair distribution will be great challenges to public health in the coming decades.


Chronic thalamic stimulation with three-dimensional MR stereotactic guidance.
To report a method of electrode implantation in the ventralis intermedius nucleus of the thalamus for the treatment of tremor using a 3-D stereotactic MR imaging technique. Five patients (three men and two women; mean age, 59 years) with medically refractory tremor had intrathalamic implantation of a stimulating electrode. Stereotactic MR imaging was performed on a 1.5-T unit equipped with an MR-compatible Leksell G stereotactic frame fixed to the patient's head. Calculation of the coordinates of the theoretical target was based on the coordinates of the anterior commissure, the posterior commissure, and the midline sagittal plane as determined via stereotactic MR imaging. During the surgical procedure, the best position for the stimulating electrode was determined by electrophysiological and clinical studies. Postoperative MR control studies were done in all cases to verify the position of the electrode. Stereotactic MR imaging allowed precise implantation of the stimulating electrode in all patients. Electrode stimulation produced a 90% reduction of the tremor in two patients, an 80% and 70% reduction in one patient each, and a persistent microthalamotomy-like effect in the fifth patient. Examination of the MR control studies showed that mean error in the positioning of the electrodes was 0.77 +/- 0.6 mm (mean +/- SD) in the x direction and 0.80 +/- 1.02 mm in the y direction. Although our series is relatively small, the precision achieved with stereotactic MR imaging proves that it can be used with confidence for precise functional neurosurgical procedures.


Typhoid Conjugate Vaccines: Advancing the Research and Public Health Agendas.
The disease burden of typhoid fever remains high in endemic areas in Asia and Africa, especially in children. Recent clinical trials conducted by the Typhoid Vaccine Acceleration Consortium show typhoid conjugate vaccine (TCV) to be safe, immunogenic, and efficacious at preventing blood culture-confirmed typhoid fever in African and Asian children. Pakistan, Liberia, and Zimbabwe recently introduced TCV through campaigns and routine childhood immunizations, providing protection for this vulnerable population. It is essential to continue this momentum while simultaneously filling data gaps-including typhoid complications-to inform decision-making on TCV introduction. A multidisciplinary approach including surveillance, water, sanitation, and hygiene investments, and large-scale TCV introduction is needed to decrease the burden and mortality of typhoid fever.


Forensic handwriting examiners' opinions on the process of production of disguised and simulated signatures.
Large-scale blind testing of forensic handwriting examiners (FHEs) has shown that authorship opinions on disguised and simulated signatures attract higher misleading and inconclusive rates than genuine signatures do. To test whether this is due to the failure of FHEs to detect the indicators of disguise/simulation behaviours we examined their opinions regarding the 'process of production' (which in this case was a choice between written naturally or written using a disguise/simulation strategy) of the questioned disguised and simulated signatures in blinded skill testing trials. The relationship between their process opinions and authorship opinions is then assessed. It was found that the majority of the inconclusive authorship opinions for both disguised and simulated signatures had a correct process opinion (707 of 1241, 57.0% for disguised; 3838 of 4368, 87.9% for simulated), with only 7.3% (90 of 1241) of the disguised and 0.85% (37 of 4368) of the simulated signatures exhibiting incorrect process opinions. For the total misleading authorship opinions relating to disguised signatures, the majority of the process opinions were correct (167 of 241, 69.3%) indicating that a disguise/simulation process was detected, but misinterpreted as being by another writer. These results show the usefulness of FHEs offering a first stage simulation/disguise process opinion without going on to form an opinion on authorship, as the support for the proposition that a signature is something other than genuine may be, in itself, of strong evidential value.


Creating a nondiffracting beam with sub-diffraction size by a phase spatial light modulator.
Due to its unique properties of nondiffracting propagation, highly-localized intensity distribution, small beam cross-section, and self-healing, a nondiffracting beam is attractive for materials processing, microscopy, and optical research. Various methods have been investigated to generate such beams with conventional optics. However, the transverse size of those nondiffracting beams is restricted by the diffraction-limit. To overcome the diffraction limit, we use the concepts of super-oscillation and the vectorial angular spectrum method to design a phase mask mirror with a focal length of 1 m, radius of 5 mm, and numerical aperture of about 0.005 for a wavelength of 632.8 nm. The phase mask mirror was created with a phase spatial light modulator. Under the illumination of a linearly polarized Gaussian wave, a nondiffracting beam was created with sub-diffraction transverse size. The maximum transverse size of the beam is smaller than the diffraction limit of 0.5λ/NA for a propagation distance greater than 43.3 mm. A nondiffracting beam with smaller transverse size can be realized by further increasing the NA value.


APP is cleaved by Bace1 in pre-synaptic vesicles and establishes a pre-synaptic interactome, via its intracellular domain, with molecular complexes that regulate pre-synaptic vesicles functions.
Amyloid Precursor Protein (APP) is a type I membrane protein that undergoes extensive processing by secretases, including BACE1. Although mutations in APP and genes that regulate processing of APP, such as PSENs and BRI2/ITM2B, cause dementias, the normal function of APP in synaptic transmission, synaptic plasticity and memory formation is poorly understood. To grasp the biochemical mechanisms underlying the function of APP in the central nervous system, it is important to first define the sub-cellular localization of APP in synapses and the synaptic interactome of APP. Using biochemical and electron microscopy approaches, we have found that APP is localized in pre-synaptic vesicles, where it is processed by Bace1. By means of a proteomic approach, we have characterized the synaptic interactome of the APP intracellular domain. We focused on this region of APP because in vivo data underline the central functional and pathological role of the intracellular domain of APP. Consistent with the expression of APP in pre-synaptic vesicles, the synaptic APP intracellular domain interactome is predominantly constituted by pre-synaptic, rather than post-synaptic, proteins. This pre-synaptic interactome of the APP intracellular domain includes proteins expressed on pre-synaptic vesicles such as the vesicular SNARE Vamp2/Vamp1 and the Ca2+ sensors Synaptotagmin-1/Synaptotagmin-2, and non-vesicular pre-synaptic proteins that regulate exocytosis, endocytosis and recycling of pre-synaptic vesicles, such as target-membrane-SNAREs (Syntaxin-1b, Syntaxin-1a, Snap25 and Snap47), Munc-18, Nsf, α/β/γ-Snaps and complexin. These data are consistent with a functional role for APP, via its carboxyl-terminal domain, in exocytosis, endocytosis and/or recycling of pre-synaptic vesicles.


The need for vigilance: the case of a false-negative newborn screen for cystic fibrosis.
Cystic fibrosis (CF) is the most common life-limiting recessive genetic disorder in the white population. CF is caused by abnormalities in the gene that codes for the cystic fibrosis transmembrane conductance regulator protein (CFTR) and may result in severe chronic lung disease, poor growth, and malnutrition. Physicians often do not consider CF in the differential diagnosis of an infant with failure to thrive in the presence of a negative newborn screening (NBS) result. In Minnesota, newborn infants are screened for CF by immunoreactive trypsinogen (IRT) testing followed by DNA analysis if the IRT screen result is abnormal. All positive NBS results are followed by confirmatory sweat-testing by pilocarpine iontophoresis. We present here the case of a 1-month-old white boy with failure to thrive, chronic diarrhea, and severe malnutrition. Minnesota state CF NBS results were negative at birth (IRT: 43 ng/mL [96% cutoff value: 52 ng/mL]). Clinical symptoms resulted in sweat-testing by Gibson-Cooke pilocarpine iontophoresis at 1 month of age, and the result was positive (102 mmol Cl(-)/L [normal: ≤30 mmol Cl(-)/L]). CFTR mutation analysis confirmed a homozygous f508del genotype, and stool pancreatic elastase testing revealed severe exocrine pancreatic insufficiency. This case represents the first known false-negative result in Minnesota since the initiation of NBS for CF in 2006, which illustrates the importance of considering CF in the evaluation of an infant with failure to thrive and symptoms of malabsorption, regardless of NBS results.


Smokers With Adequate Vitamin C Intake Show a Preferable Pulmonary Function Test.
Airflow obstruction often results from the chronic inflammation caused by cigarette smoke. It has been concluded that cigarette smoke-induced oxidative damage is prevented by ascorbic acid on a cellular level. The purpose of the current study was to explore the effect of vitamin C intake on pulmonary function in established smokers (100 or more cigarettes) and never-smokers in a Korean population. The 2974 enrolled men and women over the age of 40 in the Korea National Health and Nutrition Examination Survey IV 2008 were divided into 4 groups based on smoking patterns (never-smoker vs established smoker) and vitamin C intake from dietary assessment (higher vs lower; median value: 77.18 mg/day). Univariate analysis showed associations between forced expiratory volume in 1 second (FEV1), FEV/forced vital capacity (FVC) and age, gender, body mass index (BMI), pack-years, vitamin C intake, and additional micronutrient intake. On multiple logistic regression analysis, the odds ratios (95% confidence interval) for FEV1 < 80% predicted were 1.000 (never-smokers, higher vitamin C intake), 1.067 (0.823, 1.383; never-smokers, lower vitamin C intake), 1.224 (0.871, 1.721; established smokers, higher vitamin C intake), and 1.479 (1.057, 2.072; established smokers, lower vitamin C intake). The odds ratios for FEV1/FVC < 0.70 were 1.177 (0.821, 1.687; never-smokers, lower vitamin C intake), 1.637 (1.094, 2.445; established smokers, higher vitamin C intake), and 2.093 (1.403, 3.122; established smokers, lower vitamin C intake) after adjusting for confounding factors (p < 0.001). Korean smokers with adequate vitamin C intake showed a preferable pulmonary function test.


Activity-Based Restorative Therapies after Spinal Cord Injury: Inter-institutional conceptions and perceptions.
This manuscript is a review of the theoretical and clinical concepts provided during an inter-institutional training program on Activity-Based Restorative Therapies (ABRT) and the perceptions of those in attendance. ABRT is a relatively recent high volume and intensity approach toward the restoration of neurological deficits and decreasing the risk of secondary conditions associated with paralysis after spinal cord injury (SCI). ABRT is guided by the principle of neuroplasticity and the belief that even those with chronic SCI can benefit from repeated activation of the spinal cord pathways located both above and below the level of injury. ABRT can be defined as repetitive-task specific training using weight-bearing and external facilitation of neuromuscular activation. The five key components of ABRT are weight-bearing activities, functional electrical stimulation, task-specific practice, massed practice and locomotor training which includes body weight supported treadmill walking and water treadmill training. The various components of ABRT have been shown to improve functional mobility, and reverse negative body composition changes after SCI leading to the reduction of cardiovascular and other metabolic disease risk factors. The consensus of those who received the ABRT training was that ABRT has much potential for enhancement of recovery of those with SCI. Although various institutions have their own strengths and challenges, each institution was able to initiate a modified ABRT program.


Induction of steroidogenic cells from adult stem cells and pluripotent stem cells [Review].
Steroid hormones are mainly produced in adrenal glands and gonads. Because steroid hormones play vital roles in various physiological processes, replacement of deficient steroid hormones by hormone replacement therapy (HRT) is necessary for patients with adrenal and gonadal failure. In addition to HRT, tissue regeneration using stem cells is predicted to provide novel therapy. Among various stem cell types, mesenchymal stem cells can be differentiated into steroidogenic cells following ectopic expression of nuclear receptor (NR) 5A subfamily proteins, steroidogenic factor-1 (also known as adrenal 4 binding protein) and liver receptor homolog-1, with the aid of cAMP signaling. Conversely, these approaches cannot be applied to pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, because of poor survival following cytotoxic expression of NR5A subfamily proteins. However, if pluripotent stem cells are first differentiated through mesenchymal lineage, they can also be differentiated into steroidogenic cells via NR5A subfamily protein expression. This approach offers a potential suitable cells for future regenerative medicine and gene therapy for diseases caused by steroidogenesis deficiencies. It represents a powerful tool to investigate the molecular mechanisms involved in steroidogenesis. This article highlights our own and current research on the induction of steroidogenic cells from various stem cells. We also discuss the future direction of their clinical application.


Bond strength between a silica glass-fiber-reinforced composite and artificial polymer teeth.
A resin-bonded prosthesis is preferred when conservation of abutment tooth structure is required, although a problem with such systems is bonding between the fixed partial denture material and a polymer pontic. The aim of this study was to investigate the bond strength of a silica glass-fiber-reinforced composite to polymer teeth compared with the bond strength of a proprietary heat-polymerized denture base polymer and also between a layer of the polymerized matrix polymer of the experimental base material and an additional layer of the same material. Polymeric artificial maxillary incisors and two heat-polymerized base materials (ProBase Hot and an experimental formulation) were used. Three test groups (each n=6) were prepared: bonding between artificial tooth and an experimental composite (group-TC), bonding between tooth and ProBase Hot (group-TP), and bonding between experimental base material and experimental base material (group-BB). The content of inorganic particles of teeth was determined by combustion analysis. The bond strengths were measured in shear with a universal testing machine and the data were analyzed by one-way ANOVA and post-hoc Scheffé test (p<0.05). After failure, the fracture surfaces were examined by optical microscopy. The content of inorganic particles in the teeth was low. There is no statistically significant difference in bond strength between the groups. Bond strength of the experimental composite to artificial polymer teeth was as good as that of a conventional heat-polymerized denture base polymer. The strong bonding between the experimental material and artificial teeth makes this combination suitable for resin-bonded prostheses.


Neuronal populations in the occipital cortex of the blind synchronize to the temporal dynamics of speech.
The occipital cortex of early blind individuals (EB) activates during speech processing, challenging the notion of a hard-wired neurobiology of language. But, at what stage of speech processing do occipital regions participate in EB? Here we demonstrate that parieto-occipital regions in EB enhance their synchronization to acoustic fluctuations in human speech in the theta-range (corresponding to syllabic rate), irrespective of speech intelligibility. Crucially, enhanced synchronization to the intelligibility of speech was selectively observed in primary visual cortex in EB, suggesting that this region is at the interface between speech perception and comprehension. Moreover, EB showed overall enhanced functional connectivity between temporal and occipital cortices that are sensitive to speech intelligibility and altered directionality when compared to the sighted group. These findings suggest that the occipital cortex of the blind adopts an architecture that allows the tracking of speech material, and therefore does not fully abstract from the reorganized sensory inputs it receives.


Current Status of Pediatric Critical Care in Korea: Results of 2015 National Survey.
The aim of this study was to describe the structure, organization, management, and staffing of pediatric critical care (PCC) in Korea. We directed a questionnaire survey for all Upper Grade General Hospitals (n = 43) in Korea in 2015. The first questionnaire was mainly about structure, organization, and staffing and responses were obtained from 32 hospitals. The second questionnaire was mainly about patients and management. Responses to second questionnaire were obtained from 18 hospitals. Twelve from 32 Upper Grade General Hospitals had pediatric intensive care units (PICUs) and 11 of them had the PICU which was exclusive for children. Total number of PICU beds in Korea was 113. The ratio of the number of PICU beds to the number of children was 1:77,460 in Korea and this ratio is lower than that of other developed countries. The mean number of beds in the PICUs was 9.4 ± 9.3 (range, 2-30). There were 16 medical doctors who were assigned for PCC and only 5 of them were full time pediatric intensivists. In the 18 Upper Grade General Hospitals that responded to the second questionnaire survey, there were 97 patients in the PICUs with an average number of 5.7 ± 7.2 (range, 0-22) on the survey day. The mean age of the patients was 3.4 ± 5.6 years. The mean length of hospital stay was 82 ± 271 days. The mean Pediatric Risk of Mortality score III was 9.4 ± 7.8 at the time of admission to the PICUs. There is a considerable shortage of PICU beds compared to those in developed countries. In addition, the proportion of PICUs with PCC specialists is much lower than those in the US and European countries.


OSIRIS, an entirely in-house developed drug discovery informatics system.
We present OSIRIS, an entirely in-house developed drug discovery informatics system. Its components cover all information handling aspects from compound synthesis via biological testing to preclinical development. Its design principles are platform and vendor independence, a consistent look and feel, and complete coverage of the drug discovery process by custom tailored applications. These include electronic laboratory notebook applications for biology and chemistry, tools for high-throughput and secondary screening evaluation, chemistry-aware data visualization, physicochemical property prediction, 3D-pharmacophore comparisons, interactive modeling, computing grid based ligand-protein docking, and more. Most applications are developed in Java and are built on top of a Java library layer that provides reusable cheminformatics functionality and GUI components such as chemical editors, structure canonicalization, substructure search, combinatorial enumeration, enhanced stereo perception, force field minimization, and conformation generation.


Genetic signatures of lipid metabolism evolution in Cetacea since the divergence from terrestrial ancestor.
In mammalian evolutionary history, Cetacea (whales, dolphins and porpoises) achieved astonishing success by adapting to an aquatic environment. One unique characteristic of cetaceans, contributing to this adaptive success, is efficient lipid utilization. Here, we report a comparative genetic analysis of five aquatic and five terrestrial Cetartiodactyla species using 144 genes associated with lipid metabolism. Mutation ratio (dN /dS ), amino acid substitution in functional domains and metabolic pathways were evaluated using branch-site model in PAML, Pfam and KEGG, respectively. Our tests detected 20 positively selected genes in Cetacea compared to 11 in Bovidae with little overlap between the lineages. We identified lineage-specific patterns of amino acid substitutions and functional domains that were mutually exclusive between cetaceans and bovids, supporting divergent evolution of lipid metabolism since the divergence of these taxa from a common ancestor. Moreover, a pathway analysis showed that the identified genes in cetaceans were associated with lipid digestion, lipid storage and energy-producing pathways. This study emphasizes the evolutionary context of lipid metabolism modification of cetaceans and provides a foundation for future studies of elucidating the adapted biological mechanisms of cetacean lipid metabolism and a framework for incorporating ecological context into studies aimed at investigating adaptive evolution.


Protein phosphatase 2A dephosphorylates CaBP4 and regulates CaBP4 function.
CaBP4 is a neuronal Ca(2+)-binding protein that is expressed in the retina and in the cochlea, and is essential for normal photoreceptor synaptic function. CaBP4 is phosphorylated by protein kinase C zeta (PKCζ) in the retina at serine 37, which affects its interaction with and modulation of voltage-gated Ca(v)1 Ca(2+) channels. In this study, we investigated the potential role and functional significance of protein phosphatase 2A (PP2A) in CaBP4 dephosphorylation. The effect of protein phosphatase inhibitors, light, and overexpression of PP2A subunits on CaBP4 dephosphorylation was measured in in vitro assays. Pull-down experiments using retinal or transfected HEK293 cell lysates were used to investigate the association between CaBP4 and PP2A subunits. Electrophysiologic recordings of cotransfected HEK293 cells were performed to analyze the effect of CaBP4 dephosphorylation in modulating Ca(v)1.3 currents. PP2A inhibitors, okadaic acid (OA), and fostriecin, but not PP1 selective inhibitors, NIPP-1, and inhibitor 2, block CaBP4 dephosphorylation in retinal lysates. Increased phosphatase activity in light-dependent conditions reverses phosphorylation of CaBP4 by PKCζ. In HEK293 cells, overexpression of PP2A enhances the rate of dephosphorylation of CaBP4. In addition, inhibition of protein phosphatase activity by OA increases CaBP4 phosphorylation and potentiates the modulatory effect of CaBP4 on Ca(v)1.3 Ca(2+) channels in HEK293T cells. This study provides evidence that CaBP4 is dephosphorylated by PP2A in the retina. Our findings reveal a novel role for protein phosphatases in regulating CaBP4 function in the retina, which may fine tune presynaptic Ca(2+) signals at the photoreceptor synapse.


Corticosteroids in Sepsis: An Updated Systematic Review and Meta-Analysis.
This systematic review and meta-analysis addresses the efficacy and safety of corticosteroids in critically ill patients with sepsis. We updated a comprehensive search of MEDLINE, EMBASE, CENTRAL, and LILACS, and unpublished sources for randomized controlled trials that compared any corticosteroid to placebo or no corticosteroid in critically ill children and adults with sepsis. Reviewers conducted duplicate screening of citations, data abstraction, and, using a modified Cochrane risk of bias tool, individual study risk of bias assessment. A parallel guideline committee provided input on the design and interpretation of the systematic review, including the selection of outcomes important to patients. We assessed overall certainty in evidence using Grading of Recommendations Assessment, Development and Evaluation methodology and performed all analyses using random-effect models. For subgroup analyses, we performed metaregression and considered p value less than 0.05 as significant. Forty-two randomized controlled trials including 10,194 patients proved eligible. Based on low certainty, corticosteroids may achieve a small reduction or no reduction in the relative risk of dying in the short-term (28-31 d) (relative risk, 0.93; 95% CI, 0.84-1.03; 1.8% absolute risk reduction; 95% CI, 4.1% reduction to 0.8% increase), and possibly achieve a small effect on long-term mortality (60 d to 1 yr) based on moderate certainty (relative risk, 0.94; 95% CI, 0.89-1.00; 2.2% absolute risk reduction; 95% CI, 4.1% reduction to no effect). Corticosteroids probably result in small reductions in length of stay in ICU (mean difference, -0.73 d; 95% CI, -1.78 to 0.31) and hospital (mean difference, -0.73 d; 95% CI, -2.06 to 0.60) (moderate certainty). Corticosteroids result in higher rates of shock reversal at day 7 (relative risk, 1.26; 95% CI, 1.12-1.42) and lower Sequential Organ Failure Assessment scores at day 7 (mean difference, -1.39; 95% CI, -1.88 to -0.89) (high certainty). Corticosteroids likely increase the risk of hypernatremia (relative risk, 1.64; 95% CI, 1.32-2.03) and hyperglycemia (relative risk, 1.16; 95% CI, 1.08-1.24) (moderate certainty), may increase the risk of neuromuscular weakness (relative risk, 1.21; 95% CI, 1.01-1.52) (low certainty), and appear to have no other adverse effects (low or very low certainty). Subgroup analysis did not demonstrate a credible subgroup effect on any of the outcomes of interest (p > 0.05 for all). In critically ill patients with sepsis, corticosteroids possibly result in a small reduction in mortality while also possibly increasing the risk of neuromuscular weakness.


Molecularly imprinted solid-phase extraction of diazepam and its metabolites from hair samples.
An anti-diazepam, molecularly imprinted polymer (MIP) has been synthesized and used to extract diazepam and other benzodiazepines from hair samples via a molecularly imprinted solid-phase extraction (MISPE) protocol. Optimum retention of diazepam on the MIP columns was achieved using an apolar solvent, and the binding capacity of the polymer toward diazepam was found to be 110 ng of diazepam/mg of polymer. The recovery of a 50 ng diazepam standard spiked into blank hair was 93%, with good precision (RSD = 1.5%). The LOD and LOQ of diazepam in spiked hair samples were 0.09 and 0.14 ng/mg, respectively. The MISPE method was demonstrated to be applicable to the analysis of diazepam metabolites and other benzodiazepine drugs, in addition to diazepam itself. The application of the extraction method to postmortem hair samples yielded results that were in good agreement with the corresponding ELISA data (from blood samples) and data arising from the analysis of the same blood samples using a validated in-house SPE-LC-MS-MS method.


PLM detection by actigraphy compared to polysomnography: a validation and comparison of two actigraphs.
To compare periodic leg movement (PLM) counts obtained with polysomnography (PSG) to those obtained from actigraphy with two devices (Actiwatch and PAM-RL). Twenty-four patients underwent full night actigraphy with Actiwatch from both legs and simultaneous PSG. Out of these patients, 10 had additional actigraphy with PAM-RL. Bilateral and unilateral PLM indices (PLMI) for both actigraphs were calculated for time in bed and compared to polysomnographic PLMI. Additionally, a comparison between the two different actigraphs was performed. PLMI obtained with Actiwatch were significantly lower than those obtained with PSG (21.2+/-25.6/h versus 34.4+/-30.7/h; p<0.001), whereas the PLMI from PAM-RL were significantly higher than in PSG (63.6+/-39.3/h versus 37.0+/-33.5/h; p=0.009). In direct comparison, Actiwatch gave significantly lower PLMI than the PAM-RL (p=0.005). The correlations between Actiwatch and PSG (rho=0.835, p<0.001), PAM-RL and PSG (rho=0.939, p<0.001), and Actiwatch and PAM-RL (rho=0.915, p<0.001) were significant. Unilateral actigraphy compared to standard PSG gave less consistent findings. When comparing different settings of the PAM-RL, manual threshold setting resulted in PLMI that were no longer different from PSG (p=0.074), in contrast to the default threshold setting. The Actiwatch underestimated and the PAM-RL overestimated PLMI compared to PSG. Whereas PLMI obtained with two actigraphs and PSG were highly correlated, they differed in mean values. Therefore, PSG, actigraphy and also the different actigraphs cannot be interchanged in longitudinal studies, and actigraphy should not be used for diagnostic decision making based on PLM indices. The best approximation to PSG PLMI was achieved by using manual threshold setting with the PAM-RL.


Results of clubfoot treatment after manipulation and casting using the Ponseti method: experience in Harare, Zimbabwe.
The objective of this study was to evaluate the outcomes of the Ponseti manipulation and casting method for clubfoot in a tertiary hospital in Zimbabwe and explore predictors of these outcomes. A cohort study included children with idiopathic clubfoot managed from 2011 to 2013 at Parirenyatwa Hospital. Demographic data, clinical features and treatment outcomes were extracted from clinic records. The primary outcome measure was the final Pirani score (clubfoot severity measure) after manipulation and casting. Secondary outcomes included change in Pirani score (pre-treatment to end of casting), number of casts for correction, proportion receiving tenotomy and proportion lost to follow up. A total of 218 children (337 feet) were eligible for inclusion. The median age at treatment was 8 months; 173 children (268 feet) completed casting treatment within the study period. The mean length of time for corrective treatment was 10.2 weeks (9.5-10.9 weeks). Of the 45 children who did not complete treatment, 28 were under treatment and 17 were lost to follow up. A Pirani score of 1 or less was achieved in 85% of feet. Mean Pirani score at presentation was 3.80 (SD 1.15) and post-treatment 0.80 (SD 0.56, P-value <0.0001). Severity of deformity and being male were associated with a higher (worse) final Pirani score. Severity and age over two were associated with an increase in the number of casts required to correct deformity. This case series demonstrates that the majority (80%+) of children with clubfoot can achieve a good outcome with the Ponseti manipulation and casting method.


Activation of acid-sensing ion channel 3 by lysophosphatidylcholine 16:0 mediates psychological stress-induced fibromyalgia-like pain.
Fibromyalgia is commonly considered a stress-related chronic pain disorder, and daily stressors are known triggers. However, the relation between stress and pain development remains poorly defined by clinical approaches. Also, the aetiology remains largely unknown. We used a newly developed mouse model and lipidomic approaches to probe the causation and explore the biological plausibility for how perceived stress translates into chronic non-inflammatory pain. Clinical and lipidomic investigations of fibromyalgia were conducted for human validation. Using non-painful sound stimuli as psychological stressors, we demonstrated that mice developed long-lasting non-inflammatory hyperalgesia after repeated and intermittent sound stress exposure. Elevated serum malondialdehyde level in stressed mice indicated excessive oxidative stress and lipid oxidative damage. Lipidomics revealed upregulation of lysophosphatidylcholine 16:0 (LPC16:0), a product of lipid oxidisation, in stressed mice. Intramuscular LPC16:0 injection triggered nociceptive responses and a hyperalgesic priming-like effect that caused long-lasting hypersensitivity. Pharmacological or genetic inhibition of acid-sensing ion channel 3 impeded the development of LPC16:0-induced chronic hyperalgesia. Darapladib and antioxidants could effectively alleviate the stress-induced hyperalgesia by inhibiting LPC16:0 synthesis. Clinical investigations showed that excessive oxidative stress and LPC16:0 expression also exist in patients with fibromyalgia. Moreover, LPC16:0 expression was correlated with pain symptoms in patients with high oxidative stress and disease severity. Our study provides experimental evidence for the causal effect of psychological stressors on chronic pain development. The findings identify a possible pathophysiological mechanism of stress-induced chronic non-inflammatory pain at molecular, behavioural and clinical levels that might indicate a new therapeutic approach for fibromyalgia.


CRP2, a new invadopodia actin bundling factor critically promotes breast cancer cell invasion and metastasis.
A critical process underlying cancer metastasis is the acquisition by tumor cells of an invasive phenotype. At the subcellular level, invasion is facilitated by actin-rich protrusions termed invadopodia, which direct extracellular matrix (ECM) degradation. Here, we report the identification of a new cytoskeletal component of breast cancer cell invadopodia, namely cysteine-rich protein 2 (CRP2). We found that CRP2 was not or only weakly expressed in epithelial breast cancer cells whereas it was up-regulated in mesenchymal/invasive breast cancer cells. In addition, high expression of the CRP2 encoding gene CSRP2 was associated with significantly increased risk of metastasis in basal-like breast cancer patients. CRP2 knockdown significantly reduced the invasive potential of aggressive breast cancer cells, whereas it did not impair 2D cell migration. In keeping with this, CRP2-depleted breast cancer cells exhibited a reduced capacity to promote ECM degradation, and to secrete and express MMP-9, a matrix metalloproteinase repeatedly associated with cancer progression and metastasis. In turn, ectopic expression of CRP2 in weakly invasive cells was sufficient to stimulate cell invasion. Both GFP-fused and endogenous CRP2 localized to the extended actin core of invadopodia, a structure primarily made of actin bundles. Purified recombinant CRP2 autonomously crosslinked actin filaments into thick bundles, suggesting that CRP2 contributes to the formation/maintenance of the actin core. Finally, CRP2 depletion significantly reduced the incidence of lung metastatic lesions in two xenograft mouse models of breast cancer. Collectively, our data identify CRP2 as a new cytoskeletal component of invadopodia that critically promotes breast cancer cell invasion and metastasis.