Morphing as a means of generating variation in visual medical teaching materials.
In computer-based medical education, there is frequently a need to present students with pictorial data representative of the natural variation associated with disease presentations as well as the progression of disease within an individual. Because of the difficulty in acquiring such data, image acquisition is often the most resource-intensive phase of multimedia courseware development. In light of the resource demands associated with image content, many courseware designers do not make opportune use of image data, but rely instead upon text descriptions to provide variation in content. The resulting lack of adequate pictorial content often lessens the overall impact of the courseware. To overcome constraints imposed by the difficulty in acquiring pictorial content of sufficient richness, a methodology of generating variation in visual teaching materials has been developed through the use of morphing. These techniques have general applicability in creating variation in pictorial teaching materials in a variety of image-intensive domains.


Ethical considerations of neuro-oncology trial design in the era of precision medicine.
The field of oncology is currently undergoing a paradigm shift. Advances in the understanding of tumor biology and in tumor sequencing technology have contributed to the shift towards precision medicine, the therapeutic framework of targeting the individual oncogenic changes each tumor harbors. The success of precision medicine therapies, such as targeted kinase inhibitors and immunotherapies, in other cancers have motivated studies in brain cancers. The high specificity and cost of these therapies also encourage a shift in clinical trial design away from randomized control trials towards smaller, more exclusive early phase clinical trials. While these new trials advance the clinical application of increasingly precise and individualized therapies, their design brings ethical challenges . We review the pertinent ethical considerations for clinical trials of precision medicine in neuro-oncology and discuss methods to protect patients in this new era of trial design.


Genomics of wood-degrading fungi.
Woody plants convert the energy of the sun into lignocellulosic biomass, which is an abundant substrate for bioenergy production. Fungi, especially wood decayers from the class Agaricomycetes, have evolved ways to degrade lignocellulose into its monomeric constituents, and understanding this process may facilitate the development of biofuels. Over the past decade genomics has become a powerful tool to study the Agaricomycetes. In 2004 the first sequenced genome of the white rot fungus Phanerochaete chrysosporium revealed a rich catalog of lignocellulolytic enzymes. In the decade that followed the number of genomes of Agaricomycetes grew to more than 75 and revealed a diversity of wood-decaying strategies. New technologies for high-throughput functional genomics are now needed to further study these organisms.


Multifactorial origins of heart and gut defects in nipbl-deficient zebrafish, a model of Cornelia de Lange Syndrome.
Cornelia de Lange Syndrome (CdLS) is the founding member of a class of multi-organ system birth defect syndromes termed cohesinopathies, named for the chromatin-associated protein complex cohesin, which mediates sister chromatid cohesion. Most cases of CdLS are caused by haploinsufficiency for Nipped-B-like (Nipbl), a highly conserved protein that facilitates cohesin loading. Consistent with recent evidence implicating cohesin and Nipbl in transcriptional regulation, both CdLS cell lines and tissues of Nipbl-deficient mice show changes in the expression of hundreds of genes. Nearly all such changes are modest, however--usually less than 1.5-fold--raising the intriguing possibility that, in CdLS, severe developmental defects result from the collective action of many otherwise innocuous perturbations. As a step toward testing this hypothesis, we developed a model of nipbl-deficiency in zebrafish, an organism in which we can quantitatively investigate the combinatorial effects of gene expression changes. After characterizing the structure and embryonic expression of the two zebrafish nipbl genes, we showed that morpholino knockdown of these genes produces a spectrum of specific heart and gut/visceral organ defects with similarities to those in CdLS. Analysis of nipbl morphants further revealed that, as early as gastrulation, expression of genes involved in endodermal differentiation (sox32, sox17, foxa2, and gata5) and left-right patterning (spaw, lefty2, and dnah9) is altered. Experimental manipulation of the levels of several such genes--using RNA injection or morpholino knockdown--implicated both additive and synergistic interactions in causing observed developmental defects. These findings support the view that birth defects in CdLS arise from collective effects of quantitative changes in gene expression. Interestingly, both the phenotypes and gene expression changes in nipbl morphants differed from those in mutants or morphants for genes encoding cohesin subunits, suggesting that the transcriptional functions of Nipbl cannot be ascribed simply to its role in cohesin loading.


Children in taxis: an opportunity for pediatricians and emergency physicians to save lives?
Child safety devices (infant seats, booster seats, and seat belts) are effective in curbing the risk of injury; however, there remains a pattern of parental nonuse or misuse of safety seats. The aim of this study was to assess the level of knowledge and compliance of parents with children presenting for emergency care of the National Highway and Traffic Association safety seat guidelines in private cars and taxicabs. Two hundred forty-two caregivers of children (ages range, 2 weeks to 19 years) presenting for care in the pediatric emergency department of an urban university hospital were approached to complete an interviewer-administered questionnaire, and 225 participated. The questionnaire included knowledge, attitude, and behavior questions on protective equipment for various aged children. Eleven (47.8%) of 23 children 1 year or younger were reported to use infant seats often or always while riding in private cars, compared with 8 (22.2%) of 36 children 1 year or younger were reported to their use while in taxis (P < 0.05). Seventeen (85%) of 20 children older than 8 years were reported to have used seat belts often or always in private cars versus 10 (41.7%) of 24 in taxis (P < 0.01). One hundred fifty-four (99.3%) of 155 subjects knew the National Highway Traffic Safety Administration recommended position for the safety seat for their child. Most parents believed in the efficacy of child safety seats in preventing vehicle injuries and reported they would be more likely to use safety devices if they received information on their use in the emergency department. Data from this survey show that use of safety seats is lower in taxis than in private automobiles and that this is attributable to the inconvenience of carrying these seats to and from the taxi rather than financial considerations or lack of knowledge about their effectiveness. Strategies should be sought to increase availability of child safety devices in taxicabs. The emergency department, as well as the pediatrician's office or clinic, can be a locus for an educational intervention to parents and caregivers on child passenger safety.


Androgen deprivation with salvage surgery for radiorecurrent prostate cancer: results at 5-year followup.
We studied the effect of the combination of androgen deprivation with salvage surgery in patients with radiorecurrent prostate cancer. Salvage cystoprostatectomy or radical prostatectomy was performed in 29 patients with radiorecurrent prostate cancer. Of the 29 patients 24 had been treated with neoadjuvant hormonal therapy before salvage surgery, while in 5 an initial trial of androgen deprivation had failed preoperatively. The positive surgical margin rate for all patients was 31%. Margin involvement correlated strongly with disease specific and disease-free survival. At a mean followup of 5.3 years disease specific survival was 95% in men with negative surgical margins compared with 44% in those with positive surgical margins (p = 0.002). Similarly, clinical and biochemical disease-free survival was 80% in patients with negative surgical margins, while only 44% of those with positive surgical margins remained disease-free (p = 0.05). Surgical margins were positive in 80% of the men in the androgen deprivation failure group and in 21% in the neoadjuvant hormonal therapy group (p = 0.001). The disease specific survival rate after an initial trial of androgen deprivation failed was only 20% compared with 92% after neoadjuvant hormonal therapy was given preoperatively (p = 0.001) The combination of neoadjuvant hormonal therapy with salvage surgery for radiorecurrent prostate cancer resulted in a low incidence of surgical margin involvement, which correlated strongly with disease specific and disease-free survival. Patients with radiorecurrent prostate cancer in whom an initial trial of androgen deprivation fails appear to be poor candidates for salvage prostatectomy.


Risk factors, ulcer grade and management outcome of diabetic foot ulcers in a Tropical Tertiary Care Hospital.
The objective was to determine the risk factors, ulcer grade, and management outcome of patients with diabetic foot ulcers (DFU) managed in a tropical tertiary hospital. This is a prospective observational study of all consecutive diabetes mellitus (DM) patients with DFU admitted in the University of Benin Teaching Hospital, Benin City, Nigeria over a 26-month period. Data documented included age, gender, type of DM, duration of DM, risk factors of DFU, duration of DFU, Wagner's ulcer grade, and the blood glucose at presentation and management outcome. Thirty-four (55.7%) of the 61 study subjects were females. Their mean age was 56.29±12.71 years. 85.2% had type 2 DM. 13.1% of the patients were newly diagnosed diabetic at presentation. The mean duration of DM was 7.8±6.98 years. The mean duration of DFU was 46.09±47.82 days and the casual blood glucose level at presentation was 18.41±9.31mmol/l. Risk factors of DFU included spontaneous blisters (52.46%), peripheral vascular disease (44.3%), peripheral neuropathy (42.6%), and visual impairment (21.3%). The common ulcer grades were IV (44.3%) and III (36.1%).The amputation rate was 52.2% while the mortality rate was 14.3%.The baseline ulcer grade was significantly associated with the risk of lower extremity amputation, and the odds ratio was 2.36 (95% 1.06-5.21). Spontaneous blisters, peripheral vascular disease, peripheral neuropathy,and visual impairment are common risk factors of DFUs. Many of our patients with DFUs presented with grade IV and V ulcers with the resultant high rate of lower extremity amputations (LEAs). Early presentation and treatment of DFUs will reduce LEAs.


Discovery of small-molecule inhibitors targeting the ribosomal peptidyl transferase center (PTC) of M. tuberculosis.
M. tuberculosis (Mtb) is a pathogenic bacterium that causes tuberculosis, which kills more than 1.5 million people worldwide every year. Strains resistant to available antibiotics pose a significant healthcare problem. The enormous complexity of the ribosome poses a barrier for drug discovery. We have overcome this in a tractable way by using an RNA segment that represents the peptidyl transferase center as a target. By using a novel combination of NMR transverse relaxation times (T 2) and computational chemistry approaches, we have obtained improved inhibitors of the Mtb ribosomal PTC. Two phenylthiazole derivatives were predicted by machine learning models as effective inhibitors, and this was confirmed by their IC50 values, which were significantly improved over standard antibiotic drugs.


Purification of deoxyribonucleic acid polymerase delta from calf thymus: partial characterization of physical properties.
Deoxyribonucleic acid (DNA) polymerase delta has been purified 7800-fold from calf thymus, to a specific activity of 28 000 units/mg of protein. Similar to DNA polymerase delta from bone marrow [Byrnes, J.J., Downey, K. M., Black, V. L., & So, A. G. (1976) Biochemistry 15, 2817], the calf thymus enzyme is associated with 3'- to 5'-exonuclease activity. Both DNA polymerase and 3'- to 5'-exonuclease activities copurify on hydroxylapatite, DNA-cellulose, and molecular sieve chromatography. The ratio of exonuclease activity to polymerase activity is approximately 1:12. When the most highly purified fraction is subjected to polyacrylamide gel electrophoresis under nondenaturing conditions, both DNA polymerase and exonuclease activities have the same mobility at several acrylamide gel concentrations. Isoelectric focusing experiments have shown that both activities have the same pI. These data suggest that 3'- to 5'-exonuclease activity is an intrinsic property of DNA polymerase delta. The molecular weight of the enzyme, as estimated from measurements of Stokes radius and sedimentation coefficient, is 152 000.


Daily use of high-potency cannabis is associated with more positive symptoms in first-episode psychosis patients: the EU-GEI case-control study.
Daily use of high-potency cannabis has been reported to carry a high risk for developing a psychotic disorder. However, the evidence is mixed on whether any pattern of cannabis use is associated with a particular symptomatology in first-episode psychosis (FEP) patients. We analysed data from 901 FEP patients and 1235 controls recruited across six countries, as part of the European Network of National Schizophrenia Networks Studying Gene-Environment Interactions (EU-GEI) study. We used item response modelling to estimate two bifactor models, which included general and specific dimensions of psychotic symptoms in patients and psychotic experiences in controls. The associations between these dimensions and cannabis use were evaluated using linear mixed-effects models analyses. In patients, there was a linear relationship between the positive symptom dimension and the extent of lifetime exposure to cannabis, with daily users of high-potency cannabis having the highest score (B = 0.35; 95% CI 0.14-0.56). Moreover, negative symptoms were more common among patients who never used cannabis compared with those with any pattern of use (B = -0.22; 95% CI -0.37 to -0.07). In controls, psychotic experiences were associated with current use of cannabis but not with the extent of lifetime use. Neither patients nor controls presented differences in depressive dimension related to cannabis use. Our findings provide the first large-scale evidence that FEP patients with a history of daily use of high-potency cannabis present with more positive and less negative symptoms, compared with those who never used cannabis or used low-potency types.


Identification of Symmetry-Protected Topological States on Noisy Quantum Computers.
Identifying topological properties is a major challenge because, by definition, topological states do not have a local order parameter. While a generic solution to this challenge is not available yet, a broad class of topological states, namely, symmetry-protected topological (SPT) states, can be identified by distinctive degeneracies in their entanglement spectrum. Here, we propose and realize two complementary protocols to probe these degeneracies based on, respectively, symmetry-resolved entanglement entropies and measurement-based computational algorithms. The two protocols link quantum information processing to the classification of SPT phases of matter. They invoke the creation of a cluster state and are implemented on an IBM quantum computer. The experimental findings are compared to noisy simulations, allowing us to study the stability of topological states to perturbations and noise.


Ventricular tachycardia/fibrillation early after defibrillator implantation in patients with hypertrophic cardiomyopathy is explained by a high-risk subgroup of patients.
Implantable cardioverter-defibrillator (ICD) studies in patients with coronary artery disease report higher risk of ventricular tachycardia/fibrillation (VT/VF) early post-implant, potentially related to local proarrhythmic effects of ICD leads. To characterize early and long-term risk of ICD discharge for VT/VF in a large hypertrophic cardiomyopathy (HCM) cohort. By using HCM multicenter registry data, we compared long-term risk of VT/VF subsequent to an early post-implant period (a priori defined as within 3 months of implant) between patients with or without VT/VF within 3 months after ICD implantation. Over a median follow-up of 4.3 years, 109 of 506 (22%) patients with HCM who received ICDs received at least 1 ICD discharge for VT/VF. Risk of first ICD discharge for VT/VF was highest in the first year post-implant (10.8% per person-year; 95% confidence interval 7.9-13.8) and particularly in the first 3 months (17.0% per person-year; 95% confidence interval 9.8-24.3). Patients with early VT/VF (≤3 months post-implant) were older, and more commonly had secondary prevention ICDs following cardiac arrest or systolic dysfunction (end-stage HCM with ejection fraction<50%). Only 2 of 247 (0.7%) patients with primary prevention ICDs and preserved systolic function had early VT/VF. Patients with VT/VF early post-implant (≤3 months) had more than 5-fold higher risk for future VT/VF during long-term follow-up compared with patients without early VT/VF (adjusted hazard ratio 5.4; 95% confidence interval 2.3-12.6). High-risk patients with HCM and VT/VF early after ICD implantation are particularly prone to subsequent VT/VF throughout follow-up. Early ICD interventions for VT/VF are largely confined to patients with prior cardiac arrest or systolic dysfunction and therefore more likely driven by higher arrhythmic risk rather than lead-related proarrhythmia.


Succession and interaction of surface and subsurface cyanobacterial blooms in oligotrophic/mesotrophic reservoirs: A case study in Miyun Reservoir.
Subsurface cyanobacterial blooms, are a significant source of odor problems in source water and have been recorded in many oligotrophic/mesotrophic drinking water reservoirs. In this study, we explored the key driving forces responsible for the succession between surface and subsurface cyanobacteria using ecological niche modelling based upon a case study in Miyun Reservoir, China. The results suggest a negative effect of water depth and surface light irradiance (I0) on subsurface Planktothrix sp. growth (p-values < 0.001), and a unimodal effect of surface water temperature (T0) with the optimum at 23 °C (p-value < 0.001). While the surface Microcystis spp. shows a strong positive relationship with temperature (T0; p-value < 0.001), and significant effects for the interaction between T0 and I0 (p- value < 0.01). In addition, we identified the extent and type of interaction between subsurface and surface cyanobacteria and conclude that the high irradiance surface water combined with sufficient nutrients at the pre-bloom stage are key factors responsible for the preferential growth of surface cyanobacteria, while the gradual decline of the surface cyanobacteria in post-bloom stage is associated with nutrient reduction. This decline and loss of surface populations enhanced underwater irradiance and thus promoted the growth and allowed for succession of subsurface cyanobacteria in deeper layers where the nutrient supply was still adequate. Based upon this, the growth potentials for the subsurface and surface cyanobacteria are different under different environmental conditions: the subsurface cyanobacteria have greater growth potential than surface cyanobacteria in shallow oligotrophic and deep eutrophic reservoirs during median light irrigation seasons.


Protein-tyrosine kinase activity and pp60v-src expression in whole cells measured by flow cytometry.
The expression and phosphotyrosine activity of pp60v-src were measured in the B31 avian sarcoma virus-transformed rat cell line by flow cytometry using monoclonal antibodies against pp60v-src (EB7) and phosphotyrosine (1G2). Although the immunocytochemical staining was markedly heterogeneous, binding of both antibodies was significantly greater to B31 cells than to untransformed Rat 1 cells. Binding of 1G2 to phosphotyrosine residues was specific; it was entirely inhibited by adding excess phenylphosphate but was not affected by phosphoserine or phosphothreonine. The relationship between the amount of phosphorylated tyrosine measured by our FCM technique and total cellular phosphotyrosine measured by phosphoamino acid analysis was linear in vanadate-treated BALB/c 3T3 cells. Treatment of B31 cells for 48 h with herbimycin A, a benzenoid ansamycin antibiotic, to decrease the expression and tyrosine kinase activity of pp60v-src caused reductions of 42% in anti-pp60v-src and 58% in anti-phosphotyrosine antibody immunofluorescence. DNA staining with the fluorescent dye propidium iodide showed no cell cycle specificity in the binding of either antibody. Herbimycin A also caused the transformed cell line to revert to the morphology, actin configuration, and growth behavior of untransformed cells; these changes were reversed within 12 h after removal of the drug. Flow cytometric evaluation of tyrosine kinase expression and activity was fast and easy, and the results correlated well with other measures of cell phenotype. This technique can be used to quantitate the effects of drugs on oncogenic proteins such as pp60v-src and their associated tyrosine kinase activity.


Contribution of bacterial sepsis to morbidity in infants born to HIV-infected Haitian mothers.
Haiti is a country with a heavy burden of HIV infection in childbearing women. Previous studies have shown that early infant deaths are common in children of HIV-infected women. This study was designed to define the rates of and risk factors for systemic bacterial and mycobacterial infection in such children and to identify the causative agents. A cohort of 120 children born to HIV-infected mothers between May 2001 and December 2003 were prospectively observed to 15 months of age. They received comprehensive pediatric care at the GHESKIO Centers. Children were assigned to being HIV-infected by serology, RNA detection, and/or defining clinical illnesses. Blood cultures were obtained before giving antibiotics in children who were febrile or chronically ill. Blood cultures also were obtained at selected visits on well children. The mortality rate in the first 15 months was high, 22 of 106 (207/1,000 live births) in these children. Sixteen (70%) deaths were within 6 months of birth. Fourty-eight blood cultures had clinically significant organisms of which 38 were Staphylococcus aureus. Blood cultures were more likely to be positive in symptomatic and in HIV-infected children. Despite perinatal HIV treatment, mortality in children born to HIV-infected mothers remained high. Bacteremia, particularly with Staphylococcus aureus, is a partial explanation for excess illness.


Fatty acid oxidation disorders.
Genetic disorders of mitochondrial fatty acid beta-oxidation have been recognized within the last 20 years as important causes of morbidity and mortality, highlighting the physiological significance of fatty acids as an energy source. Although the mammalian mitochondrial fatty acid-oxidizing system was recognized at the beginning of the last century, our understanding of its exact nature remains incomplete, and new components are being identified frequently. Originally described as a four-step enzymatic process located exclusively in the mitochondrial matrix, we now recognize that long-chain-specific enzymes are bound to the inner mitochondrial membrane, and some enzymes are expressed in a tissue-specific manner. Much of our new knowledge of fatty acid metabolism has come from the study of patients who were diagnosed with single-gene autosomal recessive defects, a situation that seems to be further evolving with the emergence of phenotypes determined by combinations of multiple genetic and environmental factors. This review addresses the normal process of mitochondrial fatty acid beta-oxidation and discusses the clinical, metabolic, and molecular aspects of more than 20 known inherited diseases of this pathway that have been described to date.


Long non‑coding RNA HOTTIP promotes hepatocellular carcinoma tumorigenesis and development: A comprehensive investigation based on bioinformatics, qRT‑PCR and meta‑analysis of 393 cases.
HOTTIP functions as an independent biomarker in multiple cancers. However, the role of HOTTIP in hepatocellular carcinoma (HCC) remains unclear. In this study, we sought to investigate the HOTTIP expression in HCC and normal liver. We combined quantitative reverse transcription-polymerase chain reactions (qRT‑PCR), Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA), Multi Experiment Matrix (MEM) and Oncomine database to assess the clinical role and the potential molecular mechanism of HOTTIP in HCC. Furthermore, a meta‑analysis was performed to evaluate the relationship between HOTTIP and HCC tumorigenesis and development. Additionally, bioinformatics analysis, which contained Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and network analysis, were applied to investigate the underlying functions, pathways and networks of the potential genes. HOTTIP was obviously upregulated in HCC. A statistically significant higher expression of HOTTIP was found in TNM (III +Ⅳ), age (≥60), sex (male), race (white) and cirrhosis (no) compared to the control groups (P<0.05). Furthermore, the meta‑analysis of 393 cases from multiple centers indicated that HOTTIP had high diagnostic value in HCC. Additionally, according to GO and KEGG analyses, we found that the most strongly enriched functional terms were gland development, transcription factor activity and extrinsic to membrane. Also, the HOTTIP co‑expressed genes were significantly related to PPAR signaling pathway. We speculate that HOTTIP might play a vital part in HCC via regulating various pathways, especially PPAR signaling pathway. However, the detailed mechanism should be confirmed by functional experiments.


In silico analysis of potential inhibitors of Ca(2+) activated K(+) channel blocker, Charybdotoxin-C from Leiurus quinquestriatus hebraeus through molecular docking and dynamics studies.
Charybdotoxin-C (ChTx-C), from the scorpion Leiurus, quinquestriatus hebraeus blocks the calcium-activated potassium channels and causes hyper excitability of the nervous system. Detailed understanding the structure of ChTx-C, conformational stability, and intermolecular interactions are required to select the potential inhibitors of the toxin. The structure of ChTx-C was modeled using Modeller 9v7. The amino acid residues lining the binding site were predicted and used for toxin-ligand docking studies, further, selected toxin-inhibitor complexes were studied using molecular dynamics (MD) simulations. The predicted structure has 91.7% of amino acids in the core and allowed regions of Ramachandran plot. A total of 133 analog compounds of existing drugs for scorpion bites were used for docking. As a result of docking, a list of compounds was shown good inhibiting properties with target protein. By analyzing the interactions, Ser 15, Lys 32 had significant interactions with selected ligand molecules and Val5, which may have hydrophobic interaction with the cyclic group of the ligand. MD simulation studies revealed that the conformation and intermolecular interactions of all selected toxin-inhibitor complexes were stable. The interactions of the ligand and active site amino acids were found out for the best-docked poses in turn helpful in designing potential antitoxins which may further be exploited in toxin based therapies.


The proprotein convertase PC5/6 is protective against intestinal tumorigenesis: in vivo mouse model.
The secretory basic amino acid-specific proprotein convertases (PCs) have often been associated with cancer/metastasis. By controlling the cleavage of cancer-associated proteins, PCs play key roles in multiple steps of cancer development. Most analyses of the implication of PCs in cancer/metastasis relied on the use of in vitro overexpression systems or inhibitors that can affect more than one PC. Aside from the role of furin in salivary gland tumorigenesis, no other in vivo genetic model of PC-knockout was reported in relation to cancer development. Since PC5/6 is highly expressed in the small intestine, the present study examined its in vivo role in intestinal tumorigenesis. Analysis of human intestinal tumors at various stages showed a systematic down-regulation of PC5/6 expression. Since gene inactivation of PC5/6 leads to lethality at birth, we generated mice lacking PC5/6 in enterocytes and analyzed the impact of the presence or absence of this PC in the mouse ApcMin/+ model that develops numerous adenocarcinomas along the intestinal tract. This resulted in viable mice with almost no expression of PC5/6 in small intestine, but with no overt phenotype. The data showed that by themselves ApcMin/+ tumors express lower levels of PC5/6 mRNA, and that the lack of PC5/6 in enterocytes results in a significantly higher tumor number in the duodenum, with a similar trend in other intestinal segments. Finally, the absence of PC5/6 is also associated with a premature mortality of ApcMin/+ mice. Overall, these data suggest that intestinal PC5/6 is protective towards tumorigenesis, especially in mouse duodenum, and possibly in human colon.


Machine learning for comprehensive forecasting of Alzheimer's Disease progression.
Most approaches to machine learning from electronic health data can only predict a single endpoint. The ability to simultaneously simulate dozens of patient characteristics is a crucial step towards personalized medicine for Alzheimer's Disease. Here, we use an unsupervised machine learning model called a Conditional Restricted Boltzmann Machine (CRBM) to simulate detailed patient trajectories. We use data comprising 18-month trajectories of 44 clinical variables from 1909 patients with Mild Cognitive Impairment or Alzheimer's Disease to train a model for personalized forecasting of disease progression. We simulate synthetic patient data including the evolution of each sub-component of cognitive exams, laboratory tests, and their associations with baseline clinical characteristics. Synthetic patient data generated by the CRBM accurately reflect the means, standard deviations, and correlations of each variable over time to the extent that synthetic data cannot be distinguished from actual data by a logistic regression. Moreover, our unsupervised model predicts changes in total ADAS-Cog scores with the same accuracy as specifically trained supervised models, additionally capturing the correlation structure in the components of ADAS-Cog, and identifies sub-components associated with word recall as predictive of progression.


The Prevalence of and Factors Related to Vascular Hyperintensity on T1-Weighted Imaging in Acute Ischemic Stroke.
Thrombus visualization in patients with acute ischemic stroke has been detected and reported using various imaging modalities. T1-weighted imaging (T1-WI) can depict thrombi as hyperintense signals within vessels. Moreover, in addition to thrombi, T1-WI hyperintensities in arteries may suggest arterial dissection. However, the frequency of and factors related to the T1-hyperintense vessel sign (T1-HVS) are not fully known. The aim of this study was to clarify the prevalence of and related factors for the T1-HVS in patients with acute ischemic stroke. From September 2014 through December 2015, consecutive acute ischemic stroke patients who were admitted to our stroke unit within 7 days from symptom onset were retrospectively recruited from the prospective registry. A T1-HVS was defined as the presence of a hyperintense signal, with intensity higher than surrounding brain, within the vessel lumen. Moreover, T1-HVSs were separated into filled T1-HVSs (hyperintensity fills whole vessel lumen) and non-filled T1-HVSs. The frequency of the T1-HVS and the timing of emersion and the relationship between the presence of the T1-HVS and arterial occlusion were assessed. A total of 399 patients (139 women; median age 73 years; National Institutes of Health Stroke Scale score 3) were enrolled in the present study. Of these, 327 (82%) patients had T1-WI on admission. Two hundred and sixty-seven (67%) subjects had at least one follow-up T1-WI (median 6 days after admission), and 134 (34%) cases had ≥2 follow-up T1-WI examinations. The T1-HVS was observed in 18 patients during admission; therefore, the frequency of the T1-HVS in acute ischemic stroke patients was 4.5% (95% CI 2.5-6.5%). All but one (94%) of the T1-HVSs were first observed on follow-up imaging, and the median number of days from onset to T1-HVS appearance was 9. For patients having initial major artery occlusion and follow-up MRI (n = 95), sensitivity and specificity of the T1-HVS for persistent arterial occlusion on follow-up MR angiography were 22 and 100%, respectively. T1-HVS persisted for a few months and then normalized. Although there were no significant differences between filled and non-filled T1-HVS, more patients with non-filled T1-HVS had arterial dissection (43%) than those with filled T1-HVS (9%, p = 0.245). The T1-HVS was observed in 4.5% of acute ischemic stroke patients. T1-HVSs appeared in the subacute phase in arteries with persistent occlusion and remained for a few months.


Barcode sequencing and a high-throughput assay for chronological lifespan uncover ageing-associated genes in fission yeast.
Ageing-related processes are largely conserved, with simple organisms remaining the main platform to discover and dissect new ageing-associated genes. Yeasts provide potent model systems to study cellular ageing owing their amenability to systematic functional assays under controlled conditions. Even with yeast cells, however, ageing assays can be laborious and resource-intensive. Here we present improved experimental and computational methods to study chronological lifespan in Schizosaccharomyces pombe. We decoded the barcodes for 3206 mutants of the latest gene-deletion library, enabling the parallel profiling of ~700 additional mutants compared to previous screens. We then applied a refined method of barcode sequencing (Bar-seq), addressing technical and statistical issues raised by persisting DNA in dead cells and sampling bottlenecks in aged cultures, to screen for mutants showing altered lifespan during stationary phase. This screen identified 341 long-lived mutants and 1246 short-lived mutants which point to many previously unknown ageing-associated genes, including 46 conserved but entirely uncharacterized genes. The ageing-associated genes showed coherent enrichments in processes also associated with human ageing, particularly with respect to ageing in non-proliferative brain cells. We also developed an automated colony-forming unit assay to facilitate medium- to high-throughput chronological-lifespan studies by saving time and resources compared to the traditional assay. Results from the Bar-seq screen showed good agreement with this new assay. This study provides an effective methodological platform and identifies many new ageing-associated genes as a framework for analysing cellular ageing in yeast and beyond.


Synonymous codon usage in Drosophila melanogaster: natural selection and translational accuracy.
I present evidence that natural selection biases synonymous codon usage to enhance the accuracy of protein synthesis in Drosophila melanogaster. Since the fitness cost of a translational misincorporation will depend on how the amino acid substitution affects protein function, selection for translational accuracy predicts an association between codon usage in DNA and functional constraint at the protein level. The frequency of preferred codons is significantly higher at codons conserved for amino acids than at nonconserved codons in 38 genes compared between D. melanogaster and Drosophila virilis or Drosophila pseudoobscura (Z = 5.93, P < 10(-6)). Preferred codon usage is also significantly higher in putative zinc-finger and homeodomain regions than in the rest of 28 D. melanogaster transcription factor encoding genes (Z = 8.38, P < 10(-6)). Mutational alternatives (within-gene differences in mutation rates, amino acid changes altering codon preference states, and doublet mutations at adjacent bases) do not appear to explain this association between synonymous codon usage and amino acid constraint.


Cinnamic acid derivative rare-earth dinuclear complexes and one-dimensional architectures: synthesis, characterization and magnetic properties.
Discrete and extended architectures based on the association of lanthanide ions with two derivatives of cinnamic acid (2-methoxycinnamic acid and 3-methoxycinnamic acid) have been synthesized and characterized by IR spectroscopy, elemental analyses, X-ray diffraction and magnetic measurements. The use of the 2-methoxycinnamic ligand enables the synthesis of a series of dinuclear complexes (Ce3+ (1), Nd3+ (2), Gd3+ (3), Dy3+ (4)), while the presence of 3-methoxycinnamic acid induces the formation of one-dimensional coordination polymers (Ce3+ (5), Nd3+ (6), Dy3+ (7)). Investigations of the magnetic properties of the complexes reveal that they all exhibit field-induced slow relaxation of the magnetisation, mainly induced by Raman and direct relaxation processes.


Long-term change in the bone mineral density of adults with adult onset growth hormone (GH) deficiency in response to short or long-term GH replacement therapy.
Only two previous studies have assessed the effects of long-term GH replacement therapy on bone mineral density (BMD) in patients with adult onset GH deficiency. To date no study has looked at the long-term impact on BMD after a short course (6-12 months) of GH replacement. In two groups of patients with adult onset GH deficiency we have studied BMD either (a) after 3 years of continuous GH replacement or (b) 2 years after completion of a short course of GH. An open GH therapeutic study in which patients were recruited from a previous double-blind placebo-controlled study. The BMD status of all patients was unknown to the physician and patient at the time of recruitment. Group A (n = 7, three females) all received GH replacement continuously for 3 years. Group B (n = 8, five females) included six patients who received GH replacement for 6 months and two who received GH replacement for 12 months with BMD being measured at 6-monthly intervals. Single photon absorptiometry (SPA) and later single X-ray absorptiometry (SXA) were used to measure forearm cortical BMD. Dual-energy X-ray absorptiometry (DXA) was used to measure lumbar spine, trochanteric, femoral neck and Ward's area BMD. In group A lumbar spine and trochanter BMD had increased significantly from baseline by 3.7% (DXA: median change = 0.045 g/cm2; P = 0.028) and 4.0% (DXA: median change = 0.031 g/cm2; P = 0.046), respectively. There were non-significant decreases in femoral neck (1.9%) (DXA: median change = -0.02 g/cm2; P = 0.39), Ward's area (6.5%) (DXA: median change = -0.06 g/cm2; P = 0.09) and forearm (2.6%) (SPA/SXA: median change = -0.013 g/cm2; P = 0.18). In group B, compared with baseline, only trochanter BMD changed significantly, increasing by 5.9% (DXA: median change = 0.0485 g/cm2; P = 0.049). Lumbar spine (DXA: median change = -0.001 g/cm2) Ward's area (DXA: median change = 0.0135 g/cm2), femoral neck (DXA: median change = -0.005 g/cm2) and forearm cortical (SPA/SXA; median change = -0.01 g/cm2) BMD did not change significantly (P = 0.67, P = 0.57, P = 0.86 and P = 0.31, respectively). Median percentage changes compared with baseline were -0.1%, 1.8%, -0.5% and -2.1%, respectively. From the time of completion of GH therapy however, BMD increased significantly at lumbar spine, (median change = 0.023 g/cm2), Ward's area (median change = 0.03 g/cm2) and trochanter (median change = 0.056 g/cm2) (P = 0.036, P = 0.049 and P = 0.012, respectively) but not at the femoral neck (median change = 0.017 g/cm2; P = 0.31) or forearm (median change = 0 g/cm2; P = 0.75). Long-term GH replacement therapy for three years appears to have beneficial effects on bone in patients with adult onset GH deficiency particularly at the lumbar spine and trochanter; the effects on femoral neck and forearm cortical BMD, however, are less impressive. A short course (6-12 months) of GH replacement therapy results in an increase in trochanter BMD several years later, and after an initial decline in BMD whilst on GH replacement, lumbar spine and Ward's area BMD return towards their baseline values. These results emphasize that not all types of bone and skeletal sites respond to GH therapy identically. Furthermore a short course of GH replacement over 6-12 months may result in significant changes in BMD several years later.


Heterogeneity of Research Results: A New Perspective From Which to Assess and Promote Progress in Psychological Science.
Heterogeneity emerges when multiple close or conceptual replications on the same subject produce results that vary more than expected from the sampling error. Here we argue that unexplained heterogeneity reflects a lack of coherence between the concepts applied and data observed and therefore a lack of understanding of the subject matter. Typical levels of heterogeneity thus offer a useful but neglected perspective on the levels of understanding achieved in psychological science. Focusing on continuous outcome variables, we surveyed heterogeneity in 150 meta-analyses from cognitive, organizational, and social psychology and 57 multiple close replications. Heterogeneity proved to be very high in meta-analyses, with powerful moderators being conspicuously absent. Population effects in the average meta-analysis vary from small to very large for reasons that are typically not understood. In contrast, heterogeneity was moderate in close replications. A newly identified relationship between heterogeneity and effect size allowed us to make predictions about expected heterogeneity levels. We discuss important implications for the formulation and evaluation of theories in psychology. On the basis of insights from the history and philosophy of science, we argue that the reduction of heterogeneity is important for progress in psychology and its practical applications, and we suggest changes to our collective research practice toward this end.


Repressing a repressor: gibberellin-induced rapid reduction of the RGA protein in Arabidopsis.
RGA (for repressor of ga1-3) and SPINDLY (SPY) are likely repressors of gibberellin (GA) signaling in Arabidopsis because the recessive rga and spy mutations partially suppressed the phenotype of the GA-deficient mutant ga1-3. We found that neither rga nor spy altered the GA levels in the wild-type or the ga1-3 background. However, expression of the GA biosynthetic gene GA4 was reduced 26% by the rga mutation, suggesting that partial derepression of the GA response pathway by rga resulted in the feedback inhibition of GA4 expression. The green fluorescent protein (GFP)-RGA fusion protein was localized to nuclei in transgenic Arabidopsis. This result supports the predicted function of RGA as a transcriptional regulator based on sequence analysis. Confocal microscopy and immunoblot analyses demonstrated that the levels of both the GFP-RGA fusion protein and endogenous RGA were reduced rapidly by GA treatment. Therefore, the GA signal appears to derepress the GA signaling pathway by degrading the repressor protein RGA. The effect of rga on GA4 gene expression and the effect of GA on RGA protein level allow us to identify part of the mechanism by which GA homeostasis is achieved.


Does overall diet in midlife predict future aging phenotypes? A cohort study.
The impact of diet on specific age-related diseases has been studied extensively, but few investigations have adopted a more holistic approach to determine the association of diet with overall health at older ages. We examined whether diet, assessed in midlife, using dietary patterns and adherence to the Alternative Healthy Eating Index (AHEI), is associated with aging phenotypes, identified after a mean 16-year follow-up. Data were drawn from the Whitehall II cohort study of 5350 adults (age 51.3±5.3 years, 29.4% women). Diet was assessed at baseline (1991-1993). Mortality, chronic diseases, and functioning were ascertained from hospital data, register linkage, and screenings every 5 years and were used to create 5 outcomes at follow-up: ideal aging (free of chronic conditions and high performance in physical, mental, and cognitive functioning tests; 4%), nonfatal cardiovascular event (12.7%), cardiovascular death (2.8%), noncardiovascular death (7.3%), [corrected] and normal aging (73.2%). Low adherence to the AHEI was associated with an increased risk of cardiovascular and noncardiovascular death. In addition, participants with a "Western-type" diet (characterized by high intakes of fried and sweet food, processed food and red meat, refined grains, and high-fat dairy products) had lower odds of ideal aging (odds ratio for top vs bottom tertile: 0.58; 95% confidence interval, 0.36-0.94; P=.02), independently of other health behaviors. By considering healthy aging as a composite of cardiovascular, metabolic, musculoskeletal, respiratory, mental, and cognitive function, the present study offers a new perspective on the impact of diet on aging phenotypes.


A phase-synchronization and random-matrix based approach to multichannel time-series analysis with application to epilepsy.
We present a general method to analyze multichannel time series that are becoming increasingly common in many areas of science and engineering. Of particular interest is the degree of synchrony among various channels, motivated by the recognition that characterization of synchrony in a system consisting of many interacting components can provide insights into its fundamental dynamics. Often such a system is complex, high-dimensional, nonlinear, nonstationary, and noisy, rendering unlikely complete synchronization in which the dynamical variables from individual components approach each other asymptotically. Nonetheless, a weaker type of synchrony that lasts for a finite amount of time, namely, phase synchronization, can be expected. Our idea is to calculate the average phase-synchronization times from all available pairs of channels and then to construct a matrix. Due to nonlinearity and stochasticity, the matrix is effectively random. Moreover, since the diagonal elements of the matrix can be arbitrarily large, the matrix can be singular. To overcome this difficulty, we develop a random-matrix based criterion for proper choosing of the diagonal matrix elements. Monitoring of the eigenvalues and the determinant provides a powerful way to assess changes in synchrony. The method is tested using a prototype nonstationary noisy dynamical system, electroencephalogram (scalp) data from absence seizures for which enhanced cortico-thalamic synchrony is presumed, and electrocorticogram (intracranial) data from subjects having partial seizures with secondary generalization for which enhanced local synchrony is similarly presumed.


Trading stages: life expectancies in structured populations.
Interest in stage-and age structured models has recently increased because they can describe quantitative traits such as size that are left out of age-only demography. Available methods for the analysis of effects of vital rates on lifespan in stage-structured models have not been widely applied because they are hard to use and interpret, and tools for age and stage structured populations are missing. We present easily interpretable expressions for the sensitivities and elasticities of life expectancy to vital rates in age-stage models, and illustrate their application with two biological examples. Much of our approach relies on trading of time and mortality risk in one stage for time and risk in others. Our approach contributes to the new framework of the study of age- and stage-structured biodemography.


Comparison between a generalized Newtonian model and a network-type multiscale model for hemodynamic behavior in the aortic arch: Validation with 4D MRI data for a case study.
Blood is a complex fluid in which the presence of the various constituents leads to significant changes in its rheological properties. Thus, an appropriate non-Newtonian model is advisable; and we choose a Modified version of the rheological model of Phan-Thien and Tanner (MPTT). The different parameters of this model, derived from the rheology of polymers, allow characterization of the non-Newtonian nature of blood, taking into account the behavior of red blood cells in plasma. Using the MPTT model that we implemented in the open access software OpenFOAM, numerical simulations have been performed on blood flow in the thoracic aorta for a healthy patient. We started from a patient-specific model which was constructed from medical images. Exiting flow boundary conditions have been developped, based on a 3-element Windkessel model to approximate physiological conditions. The parameters of the Windkessel model were calibrated with in vivo measurements of flow rate and pressure. The influence of the selected viscosity of red blood cells on the flow and wall shear stress (WSS) was investigated. Results obtained from this model were compared to those of the Newtonian model, and to those of a generalized Newtonian model, as well as to in vivo dynamic data from 4D MRI during a cardiac cycle. Upon evaluating the results, the MPTT model shows better agreement with the MRI data during the systolic and diastolic phases than the Newtonian or generalized Newtonian model, which confirms our interest in using a complex viscoelastic model.


IMPAIRED EXECUTIVE FUNCTIONING IN PEDIATRIC TRICHOTILLOMANIA (HAIR PULLING DISORDER).
No neurocognitive examinations of pediatric trichotillomania (hair pulling disorder; HPD) have taken place. As a result, science's understanding of the underlying pathophysiology associated with HPD in youths is greatly lacking. The present study seeks to begin to address this gap in the literature via examination of executive functioning in a stimulant-free sample of children with HPD. Sixteen and 23 children between 9 and 17 years of age meeting DSM-5 diagnostic criteria for HPD or classified as a healthy control, respectively, were recruited (N = 39) to complete structured interviews, self-reports, and a subset of tests from the Cambridge Automatic Neurocognitive Test Assessment Battery (CANTAB) assessing cognitive flexibility/reversal learning (intradimensional/extradimensional; IED), working memory (spatial span; SSP), and planning and organization (Stocking of Cambridge; SOC). Hierarchical regression analyses indicated that, after controlling for appropriate covariates, diagnostic status predicted impaired performance on both the IED (reversal learning only) and SOC (planning and organization) but failed to predict cognitive flexibility or working memory capacity. Correlational analyses revealed that pulling severity was strongly related to working memory capacity, while disparate relationships between pulling styles (automatic, focused pulling) were evident with respect to working memory and planning and organization. Children with HPD performed more poorly on tasks of executive functioning as compared to controls. Correlational analyses suggest potentially distinct pathophysiology underlying automatic and focused pulling warranting further research. Limitations and future areas of inquiry are discussed.


Migration, isolation and hybridization in island crop populations: the case of Madagascar rice.
Understanding how crop species spread and are introduced to new areas provides insights into the nature of species range expansions. The domesticated species Oryza sativa or Asian rice is one of the key domesticated crop species in the world. The island of Madagascar off the coast of East Africa was one of the last major Old World areas of introduction of rice after the domestication of this crop species and before extensive historical global trade in this crop. Asian rice was introduced in Madagascar from India, the Malay Peninsula and Indonesia approximately 800-1400 years ago. Studies of domestication traits characteristic of the two independently domesticated Asian rice subspecies, indica and tropical japonica, suggest two major waves of migrations into Madagascar. A population genetic analysis of rice in Madagascar using sequence data from 53 gene fragments provided insights into the dynamics of island founder events during the expansion of a crop species' geographic range and introduction to novel agro-ecological environments. We observed a significant decrease in genetic diversity in rice from Madagascar when compared to those in Asia, likely the result of a bottleneck on the island. We also found a high frequency of a unique indica type in Madagascar that shows clear population differentiation from most of the sampled Asian landraces, as well as differential exchange of alleles between Asia and Madagascar populations of the tropical japonica subspecies. Finally, despite partial reproductive isolation between japonica and indica, there was evidence of indica/japonica recombination resulting from their hybridization on the island.


Phosphorylation inactivation of endothelial nitric oxide synthesis in pulmonary arterial hypertension.
The impairment of vasodilator nitric oxide (NO) production is well accepted as a typical marker of endothelial dysfunction in vascular diseases, including in the pathophysiology of pulmonary arterial hypertension (PAH), but the molecular mechanisms accounting for loss of NO production are unknown. We hypothesized that low NO production by pulmonary arterial endothelial cells in PAH is due to inactivation of NO synthase (eNOS) by aberrant phosphorylation of the protein. To test the hypothesis, we evaluated eNOS levels, dimerization, and phosphorylation in the vascular endothelial cells and lungs of patients with PAH compared with controls. In mechanistic studies, eNOS activity in endothelial cells in PAH lungs was found to be inhibited due to phosphorylation at T495. Evidence pointed to greater phosphorylation/activation of protein kinase C (PKC) α and its greater association with eNOS as the source of greater phosphorylation at T495. The presence of greater amounts of pT495-eNOS in plexiform lesions in lungs of patients with PAH confirmed the pathobiological mechanism in vivo. Transfection of the activating mutation of eNOS (T495A/S1177D) restored NO production in PAH cells. Pharmacological blockade of PKC activity by β-blocker also restored NO formation by PAH cells, identifying one mechanism by which β-blockers may benefit PAH and cardiovascular diseases through recovery of endothelial functions.


Local temporal variability reflects functional integration in the human brain.
Local moment-to-moment variability exists at every level of neural organization, but its driving forces remain opaque. Inspired by animal work demonstrating that local temporal variability may reflect synaptic input rather than locally-generated "noise," we used publicly-available high-temporal-resolution fMRI data (N = 100 adults; 33 males) to test in humans whether greater BOLD signal variability in local brain regions was associated with functional integration (estimated via spatiotemporal PCA dimensionality). Using a multivariate partial least squares analysis, we indeed found that individuals with higher local temporal variability had a more integrated (lower dimensional) network fingerprint. Notably, temporal variability in the thalamus showed the strongest negative association with PCA dimensionality. Previous animal work also shows that local variability may upregulate from thalamus to visual cortex; however, such principled upregulation from thalamus to cortex has not been demonstrated in humans. In the current study, we rather establish a more general putative dynamic role of the thalamus by demonstrating that greater within-person thalamo-cortical upregulation in variability is itself a unique hallmark of greater functional integration that cannot be accounted for by local fluctuations in several other well-known integrative-hub regions. Our findings indicate that local variability primarily reflects functional integration, and establish a fundamental role for the thalamus in how the brain fluctuates and communicates across moments.


An observational study of monitoring of vital signs in children admitted to Kenyan hospitals: an insight into the quality of nursing care?
Measurement and correct interpretation of vital signs is part of routine clinical care. Repeated measurement enhances early recognition of deterioration, may help prevent morbidity and mortality and is a standard of care in most countries. To examine documentation of vital signs by clinicians for admissions to paediatric wards in Kenyan hospitals, to describe monitoring frequency by nurses and explore factors influencing frequency. Vital signs information (temperature, respiratory and pulse rate) for the first 48 hours of admission was collected from case records of children admitted with non-surgical conditions to 13 Kenyan county hospitals between September 2013 and April 2016. A mixed effect negative binomial regression model was used to explore whether the severity of illness (indicated by danger signs or severe diagnostic episodes) is associated with increased vital signs observation frequency. We examined 54 800 admission episodes with an overall mortality 6.1%. Nurse to bed ratios were very low (1:10 to 1:41 across hospitals). Admitting clinicians documented all or no vital signs in 57.0% and 8.4% cases respectively. For respiratory and pulse rates there was pronounced even end-digit preference (an indicator of incorrect information) and high frequency recording of specific values (P < 0.001) suggesting approximation. Monitoring frequency was explored in 41 738 children. Those with inpatient stays ≥48 hours were expected to have a vital signs count of 18, hospitals varied but most did not achieve this benchmark (median 9, range 2-30). There were clinically small but significant associations between vital signs count and presence of multiple severe illnesses or presence of severe pallor (adjusted relative risk ratio = 1.04, P < 0.01, 95% confidence interval CI = 1.02-1.06 and 1.05, P = 0.02, 95% CI = 1.01-1.09, respectively). Data suggest accurate admission measures are sometimes missing especially for pulse and respiratory rates, possibly linked to manual measurement. Monitoring frequency is often low in the high risk population studied probably indicating how quality of nursing care is undermined by considerable human resource shortages.


Flagellar radial spoke protein 2 is a calmodulin binding protein required for motility in Chlamydomonas reinhardtii.
Genetic and morphological studies have revealed that the radial spokes regulate ciliary and flagellar bending. Functional and biochemical analysis and the discovery of calmodulin in the radial spokes suggest that the regulatory mechanism involves control of axonemal protein phosphorylation and calcium binding to spoke proteins. To identify potential regulatory proteins in the radial spoke, in-gel kinase assays were performed on isolated axonemes and radial spoke fractions. The results indicated that radial spoke protein 2 (RSP2) can bind ATP and transfer phosphate in vitro. RSP2 was cloned and mapped to the PF24 locus, a gene required for motility. Sequencing revealed that pf24 contains a point mutation converting the first ATG to ATA, resulting in only trace amounts of RSP2 and confirming the RSP2 mapping. Surprisingly, the sequence does not include signature domains for conventional kinases, indicating that RSP2 may not perform as a protein kinase in vivo. However, the predicted RSP2 protein sequence contains Ca2+-dependent calmodulin binding motifs and a GAF domain, a domain found in diverse signaling proteins for binding small ligands including cyclic nucleotides. As predicted from the sequence, recombinant RSP2 binds calmodulin in a calcium-dependent manner. We postulate that RSP2 is a regulatory subunit of the radial spoke involved in localization of calmodulin for control of motility.


Bile Acid Physiology.
The primary bile acids (BAs) are synthetized from colesterol in the liver, conjugated to glycine or taurine to increase their solubility, secreted into bile, concentrated in the gallbladder during fasting, and expelled in the intestine in response to dietary fat, as well as bio-transformed in the colon to the secondary BAs by the gut microbiota, reabsorbed in the ileum and colon back to the liver, and minimally lost in the feces. BAs in the intestine not only regulate the digestion and absorption of cholesterol, triglycerides, and fat-soluble vitamins, but also play a key role as signaling molecules in modulating epithelial cell proliferation, gene expression, and lipid and glucose metabolism by activating farnesoid X receptor (FXR) and G-protein-coupled bile acid receptor-1 (GPBAR-1, also known as TGR5) in the liver, intestine, muscle and brown adipose tissue. Recent studies have revealed the metabolic pathways of FXR and GPBAR-1 involved in the biosynthesis and enterohepatic circulation of BAs and their functions as signaling molecules on lipid and glucose metabolism.


COL1-related overlap disorder: A novel connective tissue disorder incorporating the osteogenesis imperfecta/Ehlers-Danlos syndrome overlap.
The 2017 classification of Ehlers-Danlos syndromes (EDS) identifies three types associated with causative variants in COL1A1/COL1A2 and distinct from osteogenesis imperfecta (OI). Previously, patients have been described with variable features of both disorders, and causative variants in COL1A1/COL1A2; but this phenotype has not been included in the current classification. Here, we expand and re-define this OI/EDS overlap as a missing EDS type. Twenty-one individuals from 13 families were reported, in whom COL1A1/COL1A2 variants were found after a suspicion of EDS. None of them could be classified as affected by OI or by any of the three recognized EDS variants associated with COL1A1/COL1A2. This phenotype is dominated by EDS-related features. OI-related features were limited to mildly reduced bone mass, occasional fractures and short stature. Eight COL1A1/COL1A2 variants were novel and five recurrent with a predominance of glycine substitutions affecting residues within the procollagen N-proteinase cleavage site of α1(I) and α2(I) procollagens. Selected variants were investigated by biochemical, ultrastructural and immunofluorescence studies. The pattern of observed changes in the dermis and in vitro for selected variants was more typical of EDS rather than OI. Our findings indicate the existence of a wider recognizable spectrum associated with COL1A1/COL1A2.


[Cross-sectoral Approach of a Perioperative Management Center for General Thoracic Surgery].
Perioperative assessment and care, such as enhanced recovery after surgery (ERAS), is very important for improving the clinical outcomes of patients who have undergone surgery. However, professional assessments and care cannot be achieved through the actions of only 1 surgical department. We established a perioperative management center(PERIO) comprised of surgeons, dedicated nurses, anesthesiologists, dentists, physiotherapists, pharmacists, and nutritionists to perform intensive cross-sectoral perioperative management. In this manuscript, we investigated the impact of PERIO on the clinical outcomes of 127 elderly patients who underwent thoracic surgery for the resection of non-small cell lung cancer (NSCLC). We categorized these 127 patients into 3 groups:① those treated before the introduction of PERIO (between January 2006 to August 2008), ② those treated during the early phase after PERIO introduction (September 2008 to December 2011), and ③ those treated during the late phase after PERIO introduction( January 2012 to December 2014). Radical operations were performed significantly more frequently after PERIO introduction than before PERIO introduction, while the postoperative complication rates were similar among the 3 groups. The duration of postoperative hospitalization was reduced after the introduction of PERIO, and the hospital surplus increased after the introduction of PERIO. In conclusion, PERIO may play an important role in improving the clinical outcomes of thoracic surgery, especially for elderly patients with NSCLC.


Human Cerebrospinal Fluid Induces Neuronal Excitability Changes in Resected Human Neocortical and Hippocampal Brain Slices.
Human cerebrospinal fluid (hCSF) has proven advantageous over conventional medium for culturing both rodent and human brain tissue. In addition, increased activity and synchrony, closer to the dynamic states exclusively recorded in vivo, were reported in rodent slices and cell cultures switching from artificial cerebrospinal fluid (aCSF) to hCSF. This indicates that hCSF possesses properties that are not matched by the aCSF, which is generally used for most electrophysiological recordings. To evaluate the possible significance of using hCSF as an electrophysiological recording medium, also for human brain tissue, we compared the network and single-cell firing properties of human brain slice cultures during perfusion with hCSF and aCSF. For measuring the overall activity from a majority of neurons within neocortical and hippocampal human slices, we used a microelectrode array (MEA) recording technique with 252 electrodes covering an area of 3.2 × 3.2 mm2. A second CMOS-based MEA with 4225 sensors on a 2 × 2 mm2 area was used for detailed mapping of action potential waveforms and cell identification. We found that hCSF increased the number of active electrodes and neurons and the firing rate of the neurons in the slices and induced an increase in the numbers of single channel and population bursts. Interestingly, not only an increase in the overall activity in the slices was observed, but a reconfiguration of the network could also be detected with specific activation and inactivation of subpopulations of neuronal ensembles. In conclusion, hCSF is an important component to consider for future human brain slice studies, especially for experiments designed to mimic parts of physiology and disease observed in vivo.


Acute life threatening events among infants on home oxygen.
Chronic lung disease (CLD) of prematurity is associated with significant morbidity, and infants discharged home in oxygen are particularly vulnerable. To assess the incidence of acute life threatening events (ALTEs) during 12-month follow-up of CLD infants discharged home receiving supplemental oxygen. All infants discharged over a one-year period were studied. Pre-discharge oxygen requirements were set on clinical grounds. Before discharge, oxygen saturations were recorded blind using a data-logger. Infant's oxygen requirements, hospital attendances, and details of ALTEs were recorded over the subsequent year. Sixteen infants were studied. Median (range): birth-weight 938 grams (448 - 1,638); gestational age 28 weeks (24 - 32); discharge oxygen requirement 0.20 litres/minute (0.05 - 0.50). Eight infants subsequently had one or more ALTEs. Discharge oxygen saturation profiles were significantly lower in these infant when compared to those not having ALTEs (p < 0.05), despite receiving supplementary oxygen. Before discharge home, formal oxygen saturation studies should be performed in infants receiving supplementary oxygen to ensure optimum oxygen delivery.


The assessment of immediate post-operative scoliosis correction using pedicle screw system by utilising the fulcrum bending technique.
Assessment of the curve flexibility is a crucial step in a surgeon's pre-operative planning for scoliosis surgery. Many techniques have been described. These include traction films, supine side bending films, push prone techniques, traction under general anaesthesia as well as fulcrum bending film. In this study, we studied the pre- and immediate post-operative radiographs of twenty eight adolescent idiopathic scoliosis (AIS) patients who were corrected using pedicle screw systems between January 2004 and August 2006. There were twenty two females and six male patients. The mean age of the patients were 17.5 years with a range of 12 to 38 years. Skeletal maturity of the patients was assessed by Risser's score. The majority was Risser 4 (15 cases, 53.6%). Based on King and Moe's classification, the most common curve was type 3 curve (15 cases, 53.6%). Among the twenty eight patients, twenty three patients underwent only posterior correction, while 5 patients underwent additional anterior release surgery. The mean pre-operative Cobb's angle for the posterior surgery group was 65.5 +/- 13.9 degrees and the mean post-operative Cobb's angle was 32.9 +/- 12.6 degrees. There was no difference between the mean correction estimated by fulcrum bending films (Fulcrum Flexibility) and the post- operative Correction Rate figures (44.2% vs. 49.9%). The mean Fulcrum Bending Correction Index (FBCI) in this group of patients is 112.8%. In the group of patients who underwent additional anterior release, their curves were noted to be larger and less flexible with the mean pre-operative Cobb's angle and Fulcrum Flexibility of 90.4 degrees +/- 9.3 degrees and 23.4% respectively. The Fulcrum Bending Correction Index (FBCI) for this group of patients was significantly higher than the posterior surgery group: i.e. 164.0% vs 112.8%. Thus, anterior release does help to improve the correction significantly. The fulcrum bending films give good pre-operative estimation of the amount of correction to be expected post-operatively. The fulcrum bending films can help to identify the curve types which might require anterior release in order to improve the scoliosis correction. Using the Fulcrum Bending Correction Index (FBCI) will also enable surgeons to quantify more accurately the amount of correction achieved by taking into account the inherent flexibility of the spine.


Phylogeography of Schisandra chinensis (Magnoliaceae) Reveal Multiple Refugia With Ample Gene Flow in Northeast China.
Temperate conifers and broadleaved mixed forests in northeast China are ideal to investigate the genetic consequences of climate changes during the last glacial maximum (LGM), 29 - 16 kya. As previous studies were focused on tree species with long generation time; here, the evolutionary history of Schisandra chinensis, a climber species with a generation time of five years, was investigated using chloroplast DNA (cpDNA), nuclear single copy gene (nSCG), and nuclear single sequence repeats (nSSRs, i.e., microsatellite) markers, along with ecological niche modeling (ENM), which predicted a suitable habitat in Korea Peninsula (KP) during the LGM. Private haplotypes and high genetic diversity of both cpDNA and nSCG were mainly found in KP and Changbai Mt. (CB). Although no significant phylogeographic structure was detected in the cpDNA and nSCG, three nSSRs clusters roughly distributed in west (CB and KP), east (north China), and north (Xiaoxing'an Range, XR) regions were found in Structure analysis. The approximate Bayesian computation analysis showed the west cluster diverged at 35.45 kya, and the other two clusters at 19.85 kya. The genetic diversity calculated for each of the three markers showed no significant correlation with latitude. Genetic differentiation of nSSRs was also not correlated with geographic distance. Migrate analysis estimated extensive gene flow between almost all genetic cluster pairs and BOTTLENECK analysis showed that few populations experienced severe bottlenecks. Overall, results indicate that S. chinensis survived the LGM in situ in multiple refugia, which likely include two macrorefugia (KP and CB) and two microrefugia (XR and north China). Extensive postglacial gene flow among the three nSSRs clusters led to uniformly distributed genetic diversity and low genetic differentiation.


A Case of HIV Infection with Thrombocytopenia: Assosiation of HIV, Thrombotic Thrombocytopenic Purpura and Brucellosis.
To report a case of HIV infection presenting with thrombotic thrombocytopenic purpura (TTP) and brucellosis that responded well to plasmapheresis and anti-infective therapy. A 64-year-old woman with moderate confusion, fever and pancytopenia was admitted. HIV infection history was taken from her family and she was not receiving antiretroviral therapy last one year. She had generalized purpuric skin lesions. Wright tube agglutination test was found positive with a 1:160 dilution and the patient was diagnosed as brucellosis. Detailed literature search showed brucellosis as a possible cause of TTP. Patient was treated by plasma exchange/fresh frozen plasma and antimicrobials and the response was excellent. Although brucellosis seems to explain the clinical picture of this patient, it is revealed that broad differential diagnosis is needed to reach uncommon diagnosis like TTP particularly in HIV infected patients.


Carbamate and organophosphate resistance in cotton pests in India, 1995 to 1999.
Monitoring for organophosphate and carbamate resistance was carried out on five major insect pests of cotton collected from 22 cotton-growing districts across India. Resistance was monitored in Helicoverpa armigera (Hübner) and Pectinophora gossypiella (Saunders) for the period 1995-1999 and for Spodoptera litura (Fabricius), Earias vittella (Fabricius) and Bemisia tabaci (Gennadius) in a survey conducted during the 1997-98 cropping season. Of the 53 field strains of H. armigera, only four were found to exhibit resistance to quinalphos, the highest 15-fold, whereas all 16 field strains tested were found to be resistant to monocrotophos. Similarly, out of 40 field strains tested, only eight were found to express appreciable resistance to methomyl. Resistance in P. gossypiella to quinalphos was high in the majority of the strains tested. Of the seven strains of E. vittella tested, two strains from northern India exhibited > 70-fold resistance to monocrotophos. Of the 11 S. litura strains tested, only four were found to exhibit resistance factors of 10 to 30-fold to quinalphos and monocrotophos. All of the B. tabaci field strains exhibited resistance to methomyl and monocrotophos and susceptibility to triazophos. Practical implications for pest control resulting from the observed patterns of cross-resistance between quinalphos, monocrotophos and methomyl are discussed.


Endoplasmic reticulum protein ERp46 in renal cell carcinoma.
An established inverse clinical correlation between serum adiponectin levels and renal cell carcinoma (RCC) aggressiveness exists. We have recently demonstrated that adiponectin suppresses clear cell RCC (ccRCC) progression through interaction with its receptor, adiponectin receptor 1 (AdipoR1). ERp46 has been shown to inhibit adiponectin signaling via interaction with AdipoR1 in HeLa cells. However, the expression of ERp46 in RCC has not been described thus far. The objectives of this study were to investigate ERp46 in RCC, its expression, its effects on RCC growth in a mouse model and whether it interacts with AdipoR1. We demonstrated a higher ERp46/AdipoR1 expression ratio in metastatic compared to non-metastatic ccRCC, as determined by immunohistochemistry of tissue microarrays and subsequent image analysis. When ERp46 was stably knocked down using shRNA or overexpressed in murine RCC RAG cells, RCC growth after subcutaneous injection in BALB/c nude mice was inhibited and accelerated, respectively. In vitro analysis to determine the molecular interaction between AdipoR1 and ERp46 included co-immunoprecipitation using human ccRCC 786-O cells and a bacterial adenylate cyclase-based two hybrid system and demonstrated no sustained AdipoR1-ERp46 interaction. This is the first report to suggest a role for ERp46 as a potential therapeutic target in RCC given its expression profile in human RCC samples and its effect on in vivo RCC growth. Since a stable interaction with AdipoR1 could not be established, we suggest that the tumorigenic properties of ERp46 in RCC cells are not related to an inhibitory modulation of AdipoR1.


Actin filaments accumulated in the nucleus remain in the vicinity of condensing chromosomes in the zebrafish early embryo.
In the cytoplasm, filamentous actin (F-actin) plays a critical role in cell regulation, including cell migration, stress fiber formation, and cytokinesis. Recent studies have shown that actin filaments that form in the nucleus are associated with diverse functions. Here, using live imaging of an F-actin-specific probe, superfolder GFP-tagged utrophin (UtrCH-sfGFP), we demonstrated the dynamics of nuclear actin in zebrafish (Danio rerio) embryos. In early zebrafish embryos up to around the high stage, UtrCH-sfGFP increasingly accumulated in nuclei during the interphase and reached a peak during the prophase. After nuclear envelope breakdown (NEBD), patches of UtrCH-sfGFP remained in the vicinity of condensing chromosomes during the prometaphase to metaphase. When zygotic transcription was inhibited by injecting α-amanitin, the nuclear accumulation of UtrCH-sfGFP was still observed at the sphere and dome stages, suggesting that zygotic transcription may induce a decrease in nuclear F-actin. The accumulation of F-actin in nuclei may contribute to proper mitotic progression of large cells with rapid cell cycles in zebrafish early embryos, by assisting in NEBD, chromosome congression, and/or spindle assembly.


Carbon dioxide as a green carbon source for the synthesis of carbon cages encapsulating porous silicon as high performance lithium-ion battery anodes.
Si/C composite is one of the most promising candidate materials for next-generation lithium-ion battery anodes. Herein, we demonstrate the novel structure of carbon cages encapsulating porous Si synthesized by the reaction between magnesium silicide (Mg2Si) and carbon dioxide (CO2) and subsequent acid washing. Benefitting from the in situ deposition through magnesiothermic reduction of CO2, the carbon cage seals the inner Si completely and shows higher graphitization than that obtained from the decomposition of acetylene. After removing MgO, pores are created, which can accommodate the volume change of the Si anode during the charge/discharge process. As the anode material for lithium-ion batteries, the porous Si/C electrode shows a charge capacity of ∼1124 mA h g-1 after 100 cycles with 86.4% capacity retention at the current density of 0.4 A g-1. When the current density increases to 1.6 and 3.2 A g-1, the capacity can still be maintained at ∼860 and ∼460 mA h g-1, respectively. The prominent cycling and rate performance is contributed by the built-in space for Si expansion, static carbon cages that prevent penetration of electrolyte and stabilize the solid electrolyte interface (SEI) outside, and fast charge transport by the novel structure.


The predicted antigenicity of the haemagglutinin of the 1918 Spanish influenza pandemic suggests an avian origin.
In 1982 we characterized the antigenic sites of the haemagglutinin of influenza A/PR/8/34, which is an influenza strain of the H1 subtype that was isolated from humans in 1934, by studying mutants which escaped neutralization by antibody. Four antigenic sites, namely Cb, Sa, Sb and Ca, were found to be located near the tip of the trimeric haemagglutinin spike. Based on the sequence of the haemagglutinin of the 1918 Spanish influenza, we can now specify the extent of divergence of antigenic sites of the haemagglutinin during the antigenic drift of the virus between 1918 and 1934. This divergence was much more extensive (40%) than the divergence (20%) in predicted antigenic sites between the 1918 Spanish influenza and an avian H1 subtype consensus sequence. These results support the hypothesis that the human 1918 pandemic originated from an avian virus of the H1 subtype that crossed the species barrier from birds to humans and adapted to humans, presumably by mutation and/or reassortment, shortly before 1918.


Heparin Enhances the Effects of Mesenchymal Stem Cell Transplantation in a Rabbit Model of Acute Myocardial Infarction.
Stem cell transplantation in combination with administration of bioactive compounds has shown promising resultsin treating myocardial infraction (MI). In the current study, we investigated the effect of combining mesenchymal stem cells(MSCs) transplantation with heparin into the infarcted heart rabbits. For this purpose, 35 male New Zealand white rabbitswere randomly divided into five groups: sham, MI, MI+ MSCs, MI+ heparin and MI+MSCs+ heparin. MI was induced by30 min ligation of the left anterior descending coronary artery. The animals of MSCs and MSCs +heparin groups wereinjected cell culture containing MSCs intramyocardially into the infarct area. Functional parameters of the left ventricle byechocardiography, serum levels of VEGF by enzyme-linked immunosorbent assay, size of fibrotic area by Masson'strichrome staining, evaluation of morphology by Haematoxylin-Eosin and capillary density alkaline phosphatase stainingwere compared between groups. Ejection fraction, fractional shortening and levels of VEGF significantly improved in MSCsand MSCs + heparin group (P<0.05). The fibrotic area was significantly reduced (p=0.009) in MSC + heparin treated animalsin comparison with MSCs. Number of live cells and angiogenesis were increased significantly in MSCs + heparin groups incomparison with MSCs (p< 0.05). Although injection of MSCs significantly restored normal function of fibrotic area, wefound that administration of heparin combined with MSCs to infarcted heart of animals could have better effects on LVfunctional parameters in fibrosis area and resulted in superior therapeutic outcome in enhancing neovascularization andimproving cardiac fibrosis.


Cortical Excitability and Activation of TrkB Signaling During Rebound Slow Oscillations Are Critical for Rapid Antidepressant Responses.
Rapid antidepressant effects of ketamine become most evident when its psychotomimetic effects subside, but the neurobiological basis of this "lag" remains unclear. Laughing gas (N2O), another NMDA-R (N-methyl-D-aspartate receptor) blocker, has been reported to bring antidepressant effects rapidly upon drug discontinuation. We took advantage of the exceptional pharmacokinetic properties of N2O to investigate EEG (electroencephalogram) alterations and molecular determinants of antidepressant actions during and immediately after NMDA-R blockade. Effects of the drugs on brain activity were investigated in C57BL/6 mice using quantitative EEG recordings. Western blot and qPCR were used for molecular analyses. Learned helplessness (LH) was used to assess antidepressant-like behavior. Immediate-early genes (e.g., bdnf) and phosphorylation of mitogen-activated protein kinase-markers of neuronal excitability-were upregulated during N2O exposure. Notably, phosphorylation of BDNF receptor TrkB and GSK3β (glycogen synthase kinase 3β) became regulated only gradually upon N2O discontinuation, during a brain state dominated by slow EEG activity. Subanesthetic ketamine and flurothyl-induced convulsions (reminiscent of electroconvulsive therapy) also evoked slow oscillations when their acute pharmacological effects subsided. The correlation between ongoing slow EEG oscillations and TrkB-GSK3β signaling was further strengthened utilizing medetomidine, a hypnotic-sedative agent that facilitates slow oscillations directly through the activation of α2-adrenergic autoreceptors. Medetomidine did not, however, facilitate markers of neuronal excitability or produce antidepressant-like behavioral changes in LH. Our results support a hypothesis that transient cortical excitability and the subsequent regulation of TrkB and GSK3β signaling during homeostatic emergence of slow oscillations are critical components for rapid antidepressant responses.


Invasive and non-invasive congeners show similar trait shifts between their same native and non-native ranges.
Differences in morphological or ecological traits expressed by exotic species between their native and non-native ranges are often interpreted as evidence for adaptation to new conditions in the non-native ranges. In turn this adaptation is often hypothesized to contribute to the successful invasion of these species. There is good evidence for rapid evolution by many exotic invasives, but the extent to which these evolutionary changes actually drive invasiveness is unclear. One approach to resolving the relationship between adaptive responses and successful invasion is to compare traits between populations from the native and non-native ranges for both exotic invaders and congeners that are exotic but not invasive. We compared a suite of morphological traits that are commonly tested in the literature in the context of invasion for three very closely related species of Centaurea, all of which are sympatric in the same native and non-native ranges in Europe and North America. Of these, C. solstitialis is highly invasive whereas C. calcitrapa and C. sulphurea are not. For all three species, plants from non-native populations showed similar shifts in key traits that have been identified in other studies as important putative adaptive responses to post-introduction invasion. For example, for all three species plants from populations in non-native ranges were (i) larger and (ii) produced seeds that germinated at higher rates. In fact, the non-invasive C. calcitrapa showed the strongest trait shift between ranges. Centaurea solstitialis was the only species for which plants from the non-native range increased allocation to defensive spines, and allocated proportionally less resources to reproduction, patterns contrary to what would be predicted by theory and other empirical studies to enhance invasion. Our results suggest caution when interpreting the commonly observed increase in size and reproductive capacity as factors that cause exotics to become invaders.


Type I and Type II hybrid aortic arch replacement: postoperative and mid-term outcome analysis.
Hybrid aortic arch replacement has emerged as a safe treatment modality for arch aneurysms, especially in patients of old age and with greater comorbid burden. We assessed our institutional outcomes in patients undergoing Types I and II hybrid aortic arch replacement. From 2005 to 2012, 685 patients underwent thoracic endovascular repair (TEVAR), of whom 104 had hybrid arch repair (open + endovascular approach). 47 of these patients had treatment for aortic arch aneurysm ± proximal ascending aortic aneurysm. The hybrid repair entailed aortic arch vessel debranching and concomitant/delayed antegrade ± retrograde TEVAR stent grafting of the arch. Type III patients were excluded from the analysis. Data was prospectively maintained. 28 patients had Type I repair, 8 had Type II repair, and 11 had Type III repair. Mean age was 71±8 years. Primary aortic pathology was aneurysm (81%), followed by chronic arch dissection (11%). 14% of patients required reoperative cardiac surgery. Stent graft deployment rate was 100% after arch vessel debranching. Postoperative endoleak rate was zero. Average cardiopulmonary bypass time was 215±64 minutes, with crossclamp time of 70±55 minutes, and circulatory arrest time of 50±17 minutes. Paraplegia rate was 5.5% (n=2), with stroke rate of 8% (n=3) and renal failure rate of 3% (n=1) requiring hemodialysis. In-hospital mortality was 8% (n=3). Mean length of stay was 17.2±14 days. Median follow-up was 30±21 months. Freedom from all-cause mortality was 71%, 60%, and 48% at 1, 3, and 5 years respectively. Aortic reoperation rate was 2.7% (n=1). No patient had Type I or III endoleak at follow-up. Freedom from mortality was improved in cases performed more recently (July 2008 to 2012) than during our early experience (2005 to June 2008) (81% versus 44% at 3 years, P=0.05). Hybrid aortic arch replacement can be performed with good postoperative and midterm results in a cohort of old patients with significant comorbidity. With greater experience, early and midterm outcomes continue to improve. The hybrid arch technique may represent a technical advancement in the field of aortic arch surgery.


Computational toxicology--a tool for early safety evaluation.
Although inappropriate pharmacokinetic properties were a major cause of attrition in the 1990s, safety issues are recognized as today's single largest cause of drug candidate failure. It is expected that the right balance of in vivo, in vitro and computational toxicology predictions applied as early as possible in the discovery process will help to reduce the number of safety issues. This review focuses on recent developments in computational toxicology. Direct modeling of toxic endpoints has been deceiving and hampered the wide acceptance of computer predictions. The current trend is to make simpler predictions, closer to the mechanism of action, and to follow them up with in vitro or in vivo assays as appropriate.


Purification of a recombinant oncolytic virus from clarified cell culture media by anion exchange monolith chromatography.
The use of viral vectors for vaccine, gene therapy, and oncolytic virotherapy applications has received increased attention in recent years. Large-scale purification of viral vector-based biotherapeutics still presents a significant technical challenge. Chromatography is the primary tool for the purification of biomolecules in the biotechnology industry; however, the majority of chromatography resins currently available have been designed for the purification of proteins. In contrast, convective interaction media monoliths are chromatographic supports that have been designed and successfully utilized for the purification of large biomolecules, including viruses, viruslike particles, and plasmids. We present a case study on the development of a purification method for recombinant Newcastle disease virus directly from clarified cell culture media using strong anion exchange monolith technology (CIMmultus QA, BIA Separations). Resin screening studies showed at least 10 times higher dynamic binding capacity of CIMmultus QA compared to traditional anion exchange chromatography resins. Design of experiments was used to demonstrate a robust operating window for the purification of recombinant virus directly from clarified cell culture without any further pH or conductivity adjustment of the load material. The capture step was successfully scaled up from 1 mL CIMmultus QA columns to the 8 L column scale and achieved a greater than 30-fold reduction in process volume. Compared to the load material, total host cell proteins were reduced by more than 76%, and residual host cell DNA by more than 57% in the elution pool, respectively. Direct loading of clarified cell culture onto a high-capacity monolith stationary phase makes convective flow chromatography an attractive alternative to centrifugation or TFF-based virus purification procedures.


Effects of dietary nitrate supplementation on symptoms of acute mountain sickness and basic physiological responses in a group of male adolescents during ascent to Mount Everest Base Camp.
The purpose of this study was to investigate the effects of dietary nitrate supplementation, in the form of beetroot juice, on acute mountain sickness (AMS) symptoms and physiological responses, in a group of young males trekking to Mount Everest Base Camp (EBC). Forty healthy male students (mean age (SD): 16 (1) yrs) trekked to EBC over 11 days. Following an overnight fast, each morning participants completed the Lake Louise AMS questionnaire and underwent a series of physiological tests: resting blood pressure as well as resting and exercising heart rate, respiratory rate, and peripheral oxygen saturation. The exercise test consisted of a standardised 2-min stepping protocol and measurements were taken in the last 10 s. Participants in the intervention arm of the study consumed 140 ml of concentrated beetroot juice daily, containing approximately 10 mmol of nitrate, while those in the control arm consumed 140 ml of concentrated blackcurrant cordial with negligible nitrate content. Drinks were taken for the first seven days at high altitude (days 2-8), in two equal doses; one with breakfast, and one with the evening meal. Mixed modelling revealed no significant between-groups difference in the incidence of AMS (Odds Ratio - nitrate vs. 1.16 (95% CI: 0.59; 2.29)). Physiological changes occurring during ascent to high altitude generally were not significantly different between the two groups (Model Coef (95% CI) - average difference nitrate vs. systolic blood pressure, 0.16 (-4.47; 4.79); peripheral oxygen saturation, 0.28 (-0.85; 1.41); heart rate, -0.48 (-8.47; 7.50) (Model Coef (95% CI) - relative difference nitrate vs. ventilatory rate, 0.95 (0.82; 1.08)). Modelling revealed that diastolic blood pressure was 3.37 mmHg (0.24; 6.49) higher for participants in the beetroot juice, however this difference was no larger than that found at baseline and no interaction effect was observed. Supplementation with dietary nitrate did not significantly change symptoms of AMS or alter key physiological variables, in a group of adolescent males during a high altitude trekking expedition. There was no evidence of harm from dietary nitrate supplementation in this context. Given the wide confidence intervals in all models, a larger sample size would be required to exclude a false negative result. Our data suggest that prolonged oral nitrate supplementation is safe and feasible at altitude but has little physiological or clinical effect.


Medical liability claims and lawsuits filed against the University of Texas System involving adult psychiatric patients.
A survey was conducted to investigate medical liability claims filed against the University of Texas System (the U.T. System), physicians, and/or institutions concerning care of adult psychiatric patients. There were 34 such claims filed between 1978 and 1991 that had closed by December 1991. In many of these cases, medical record documentation was determined to be inadequate. Of the 34 claims filed, only seven were settled on behalf of U.T. personnel or U.T. institutions. Of these seven settlements, the mean monetary amount paid per case on behalf of U.T. System and non-U.T. System parties was $98,678.57. The two categories with the largest number of claims were 1) cases involving suicides, and 2) cases involving adverse reactions to medication. The largest award was $350,000, paid on behalf of U.T. physicians, a private physician, and non-U.T. institutions for alleged negligent failure to diagnose Cushing's disease. Of this amount, only $150,000 was paid on behalf of U.T. physicians. Risk management strategies should be implemented to address areas of liability in psychiatry residency programs.


Human cardiac myosin light chain 4 (MYL4) mosaic expression patterns vary by sex.
Sex disparities modulate cardiac function, although the proteins and mechanisms remain to be elucidated. We recently demonstrated a mosaic pattern of protein expression in the heart for over 100 proteins. Here we investigate one of these proteins, myosin light chain 4 (MYL4), which is important for contractile functions by increasing force production. We assayed the expression pattern of MYL4 across 756 ventricular myocardial samples from 668 individuals utilizing a semi-automated Cell Profiler method on five tissue microarrays (TMAs) of cardiac tissues across a diverse set of diseases. The percentage of MYL4 positive cells was significantly higher in male subjects independently across all five TMAs, regardless of disease state (p = 8.66e-15). Higher MYL4 expression was also modestly associated with hypertrophic cardiomyopathy (p = 6.3e-04). MYL4 expression did not associate with sudden cardiac death or other cardiomyopathies. This study demonstrates a new mosaic pattern of protein expression that underlies sex disparities in the human heart.


Competencies and frameworks in interprofessional education: a comparative analysis.
Health professionals need preparation and support to work in collaborative practice teams, a requirement brought about by an aging population and increases in chronic and complex diseases. Therefore, health professions education has seen the introduction of interprofessional education (IPE) competency frameworks to provide a common lens through which disciplines can understand, describe, and implement team-based practices. Whilst an admirable aim, often this has resulted in more confusion with the introduction of varying definitions about similar constructs, particularly in relation to what IPE actually means.The authors explore the nature of the terms competency and framework, while critically appraising the concept of competency frameworks and competency-based education. They distinguish between competencies for health professions that are profession specific, those that are generic, and those that may be achieved only through IPE. Four IPE frameworks are compared to consider their similarities and differences, which ultimately influence how IPE is implemented. They are the Interprofessional Capability Framework (United Kingdom), the National Interprofessional Competency Framework (Canada), the Core Competencies for Interprofessional Collaborative Practice (United States), and the Curtin University Interprofessional Capability Framework (Australia).The authors highlight the need for further discussion about establishing a common language, strengthening ways in which academic environments work with practice environments, and improving the assessment of interprofessional competencies and teamwork, including the development of assessment tools for collaborative practice. They also argue that for IPE frameworks to be genuinely useful, they need to augment existing curricula by emphasizing outcomes that might be attained only through interprofessional activity.