The use of imepitoin (Pexion™) on fear and anxiety related problems in dogs - a case series.
Fear and anxiety based problems are common in dogs. Alongside behaviour modification programmes, a range of psychopharmacological agents may be recommended to treat such problems, but few are licensed for use in dogs and the onset of action of some can be delayed. The low affinity partial benzodiazepine receptor agonist imepitoin (Pexion™, Boehringer Ingelheim) is licensed for treating canine epilepsy, has a fast onset of action in dogs and has demonstrated anxiolytic properties in rodent models. This case series reports on the use of imepitoin in a group of dogs identified as having fear/anxiety based problems. Twenty dogs were enrolled into the study, attended a behaviour consultation and underwent routine laboratory evaluation. Nineteen dogs proceeded to be treated with imepitoin orally twice daily (starting dose approximately 10 mg/kg, with alterations as required to a maximum 30 mg/kg) alongside a patient-specific behaviour modification plan for a period of 11-19 weeks. Progress was monitored via owner report through daily diary entries and telephone follow-up every two weeks. A Positive and Negative Activation Scale (PANAS) of temperament was also completed by owners during baseline and at the end of the study. The primary outcome measure was average weekly global scores (AWG) from the owner diaries. Average weekly reaction scores (AWR) for each type of eliciting context was used as a secondary outcome. Seventeen dogs completed the trial. Treatment with imepitoin alongside a behaviour modification programme resulted in owner reported improvement with reduced AWG and reduced AWR for anxiety across a range of social and non-social eliciting contexts including noise sensitivities. Significant improvement was apparent within the first week of treatment, and further improvements seen at the 11 week review point. There was a significant reduction in negative activation (PANAS) with 76.5% of owners opting to continue imepitoin at their own expense after completion of the study. This study provides initial evidence indicating the potential value of imepitoin (Pexion™) alongside appropriate behaviour modification for the rapid alleviation of signs of fear and anxiety in dogs. Further research with a larger subject population and a placebo control would be useful to confirm the apparent efficacy reported here.


Effect of pH on endotoxin affinity for metal-ceramic alloys.
Crevicular pH may modify bacterial endotoxin affinity for high-noble metal-ceramic alloys. Porphyromonas gingivalis lipopolysaccharide (LPS) affinity for 3 metal-ceramic alloys at 3 different pH levels was compared in vitro by measuring adsorption to and release from the alloy surface. Metallographically polished disks were fabricated from Pd-Ag-Sn, Au-Pd-Ag-Sn-In, and Au-Pd-In-Ga alloys. Clean disks were placed individually into 1 mL at pH 6.5, 7.0, or 7.5 phosphate-buffered saline solution containing 0.9 endotoxin units per square millimeter tritiated LPS (n = 3 disks per alloy-pH group). The disks were incubated for 24 hours at 37 degrees C before being transferred to LPS-free buffer and incubated, again for 24 hours at 37 degrees C, to evaluate elution. This transfer continued at 24-hour intervals up to 96 hours total elution incubation. Lipopolysaccharide adsorption to and elution from disks was determined through liquid scintillation spectrometry. Adsorption data were evaluated with a 2-way analysis of variance (alpha=.05) and the post hoc Tukey honestly significant difference test. Lipopolysaccharide adsorption values ranged from 0.48 +/- 0.04 EU/mm(2) for the Au-Pd-Ag-Sn-In alloy at pH 7.5 to 0.75 +/- 0.04 EU/mm(2) for the Pd-Ag-Sn alloy at pH 6.5. Alloy type (P=.0001) and environmental pH (P=.0001) significantly influenced adsorption. Adsorption to the Pd-Ag-Sn and Au-Pd-In-Ga alloys at pH 6.5, 7.0, and 7.5 were similar and decreased with increasing pH. In contrast, adsorption to the Au-Pd-Ag-Sn-In alloy was significantly less than to other alloys at pH 6.5 but did not differ at other pH levels. Lipopolysaccharide release from the alloy surface could not be detected. P. gingivalis LPS affinity for metal-ceramic alloys was modified by environmental pH. The degree of LPS adsorption depended on the composition and surface chemistry of each alloy.


Educational Intervention to Improve Delirium Recognition by Nurses.
Delirium poses increased morbidity and mortality risks for hospitalized patients. Underrecognition by health care providers contributes to poor outcomes for patients. Little has been published about methods used to teach health care providers how to screen for delirium using the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU). To evaluate the effectiveness of a multimodal educational intervention for nurses in the medical intensive care unit to improve their knowledge and skills regarding delirium and delirium recognition. An educational intervention was done in the medical intensive care unit of an academic medical center. Effectiveness was evaluated via a quasi-experimental design and using preintervention and postintervention assessments. Procedural correctness of performing the CAM-ICU delirium screening also was measured. Nurses participated in 1 small-group session (n = 34). Fifteen sessions were conducted from June to September 2016, and assessments were completed before and after the intervention. The sample consisted of predominantly nurses with a bachelor's degree (56%) who had 1 to 5 years' experience (59%). Mean scores overall and on the knowledge subscale differed significantly (P < .001) from before to after the intervention. No correlation was found between demographic groups and score differences. Seventy-nine percent of participants used the tool correctly after the intervention. The educational intervention provided for these nurses further validated published reports of the benefits of an educational program about delirium. The content of the educational intervention should be targeted for the setting, the risk factors for the patient population in question, and the specific delirium screening tool used.


Dying at home: a qualitative study of family carers' views of support provided by GPs community staff.
Dying at home is the preference of many patients with life-limiting illness. This is often not achieved and a key factor is the availability of willing and able family carers. To elicit family carers' views about the community support that made death at home possible. Qualitative study in East Devon, North Lancashire, and Cumbria. Participants were bereaved family carers who had provided care at the end of life for patients dying at home. Semi-structured interviews were conducted 6-24 months after the death. Fifty-nine bereaved family carers were interviewed (54% response rate; 69% female). Two-thirds of the patients died from cancer with median time of home care being 5 months and for non-cancer patients the median time for home care was 30 months. An overarching theme was of continuity of care that divided into personal, organisational, and informational continuity. Large numbers and changes in care staff diluted personal continuity and failure of the GPs to visit was viewed negatively. Family carers had low expectations of informational continuity, finding information often did not transfer between secondary and primary care and other care agencies. Organisational continuity when present provided comfort and reassurance, and a sense of control. The requirement for continuity in delivering complex end-of-life care has long been acknowledged. Family carers in this study suggested that minimising the number of carers involved in care, increasing or ensuring personal continuity, and maximising the informational and organisational aspects of care could lead to a more positive experience.


The fiscal impact of informal caregiving to home care recipients in Canada: how the intensity of care influences costs and benefits to government.
The objective of this study was to estimate the annual costs and consequences of unpaid caregiving by Canadians from a government perspective. We estimated these costs both at the individual and population levels for caregivers aged 45 and older. We conducted a cost-benefit analysis where we considered the costs of unpaid caregiving to be potential losses in income tax revenues and changes in social assistance payments and the potential benefit of reduced paid care expenditures. Our costing methods were based on multivariate analyses using the 2007 General Social Survey, a cross-sectional survey of 23,404 individuals. We determined the differential probability of employment, wages, and hours worked by caregivers of varying intensity versus non-caregivers. We also used multivariate analysis to determine how receiving different intensities of unpaid care impacted both the probability of receiving paid care and the weekly hours of paid care received. At the lowest intensities of caregiving, there was a net benefit to government from caregiving, at both the individual and population levels. At the population level, the net benefit to government was estimated to be $4.4 billion for caregivers providing less than five hours of weekly care. At the highest intensity of caregiving, there was a net cost to government of $641 million. Our overall findings were robust to a number of changes applied in our sensitivity analysis. We found that the factor with the greatest impact on cost was the probability of labour force participation. As the biggest cost driver appears to be the higher likelihood of intense caregivers dropping out of the labour force, government policies that enable intense caregivers to balance caregiving with employment may help to mitigate these losses.


A probable cis-regulatory element on yeast mitochondrial DNA responsible for cAMP-mediated transcription.
Studies from this laboratory have suggested that mitochondrial (mt) transcription in yeast (Saccharomyces cerevisiae) is governed by changing cellular cAMP levels, and that the mechanism of such transcriptional regulation requires cAMP-dependent protein kinase (PKA) activity; these observations, in turn, suggest a trans-activation process for nucleotide-dependent mt transcriptional control. Here we demonstrate a sequence-specific mtDNA-phosphorylated protein interaction, a requisite part of such a control mechanism, using filter-binding and gel mobility shift assays with mt protein extracts and mtDNA from rho- strains whose retained mt genes show cAMP-sensitive expression. We demonstrate that the protein-mt DNA interaction depends on PKA activity, that it specifically involves a tripartite GC-rich sequence element on yeast mtDNA, and that it does not involve mt coding or promoter sequences. Sequence analysis indicates that the GC-rich element undergoing protein interaction is present in ten copies on the yeast mt genome, and that each copy is located 5' to a strong mt promoter; the elements appear in both orientations relative to, and at varying distances upstream from, the putatively associated mt promoter elements. The mt element shows no sequence homology to relevant nuclear cis-elements examined and is unrelated to published vertebrate mt cis-elements. Several lines of evidence and argument strongly suggest that this GC-rich element functions as the cis-regulatory sequence involved in cAMP-mediated transcriptional control in yeast mitochondria.


Evaluation of integrated care services in Catalonia: population-based and service-based real-life deployment protocols.
Comprehensive assessment of integrated care deployment constitutes a major challenge to ensure quality, sustainability and transferability of both healthcare policies and services in the transition toward a coordinated service delivery scenario. To this end, the manuscript articulates four different protocols aiming at assessing large-scale implementation of integrated care, which are being developed within the umbrella of the regional project Nextcare (2016-2019), undertaken to foster innovation in technologically-supported services for chronic multimorbid patients in Catalonia (ES) (7.5 M inhabitants). Whereas one of the assessment protocols is designed to evaluate population-based deployment of care coordination at regional level during the period 2011-2017, the other three are service-based protocols addressing: i) Home hospitalization; ii) Prehabilitation for major surgery; and, iii) Community-based interventions for frail elderly chronic patients. All three services have demonstrated efficacy and potential for health value generation. They reflect different implementation maturity levels. While full coverage of the entire urban health district of Barcelona-Esquerra (520 k inhabitants) is the main aim of home hospitalization, demonstration of sustainability at Hospital Clinic of Barcelona constitutes the core goal of the prehabilitation service. Likewise, full coverage of integrated care services addressed to frail chronic patients is aimed at the city of Badalona (216 k inhabitants). The population-based analysis, as well as the three service-based protocols, follow observational and experimental study designs using a non-randomized intervention group (integrated care) compared with a control group (usual care) with a propensity score matching method. Evaluation of cost-effectiveness of the interventions using a Quadruple aim approach is a central outcome in all protocols. Moreover, multi-criteria decision analysis is explored as an innovative method for health delivery assessment. The following additional dimensions will also be addressed: i) Determinants of sustainability and scalability of the services; ii) Assessment of the technological support; iii) Enhanced health risk assessment; and, iv) Factors modulating service transferability. The current study offers a unique opportunity to undertake a comprehensive assessment of integrated care fostering deployment of services at regional level. The study outcomes will contribute refining service workflows, improving health risk assessment and generating recommendations for service selection. NCT03130283 (date released 04/06/2018), NCT03768050 (date released 12/05/2018), NCT03767387 (date released 12/05/2018).


Psychiatric genome-wide association study analyses implicate neuronal, immune and histone pathways.
Genome-wide association studies (GWAS) of psychiatric disorders have identified multiple genetic associations with such disorders, but better methods are needed to derive the underlying biological mechanisms that these signals indicate. We sought to identify biological pathways in GWAS data from over 60,000 participants from the Psychiatric Genomics Consortium. We developed an analysis framework to rank pathways that requires only summary statistics. We combined this score across disorders to find common pathways across three adult psychiatric disorders: schizophrenia, major depression and bipolar disorder. Histone methylation processes showed the strongest association, and we also found statistically significant evidence for associations with multiple immune and neuronal signaling pathways and with the postsynaptic density. Our study indicates that risk variants for psychiatric disorders aggregate in particular biological pathways and that these pathways are frequently shared between disorders. Our results confirm known mechanisms and suggest several novel insights into the etiology of psychiatric disorders.


Insights on hydrogen-bond lifetimes in liquid and supercooled water.
We study the temperature dependence of the lifetime of geometric and geometric/energetic water hydrogen-bonds (H-bonds), down to supercooled water, through molecular dynamics. The probability and lifetime of H-bonds that break either by translational or librational motions and those of energetic broken H-bonds, along with the effects of transient broken H-bonds and transient H-bonds, are considered. We show that the fraction of transiently broken energetic H-bonds increases at low temperatures and that this energetic breakdown is caused by oxygen-oxygen electrostatic repulsions upon too small amplitude librations to disrupt geometric H-bonds. Hence, differences between geometric and energetic continuous H-bond lifetimes are associated with large H-bond energy fluctuations, in opposition to moderate geometric fluctuations, within common energetic and geometric H-bond definition thresholds. Exclusion of transient broken H-bonds and transient H-bonds leads to H-bond definition-independent mean lifetimes and activation energies, ~11 kJ/mol, consistent with the reactive flux method and experimental scattering results. Further, we show that power law decay of specific temporal H-bond lifetime probability distributions is associated with librational and translational motions that occur on the time scale (~0.1 ps) of H-bond breaking /re-forming dynamics. While our analysis is diffusion-free, the effect of diffusion on H-bond probability distributions where H-bonds are allowed to break and re-form, switching acceptors in between, is shown to result in neither exponential nor power law decay, similar to the reactive flux correlation function.


Prevalence of tobacco use in selected Johannesburg suburbs.
Tobacco smoking is estimated to kill more than 44 000 South Africans every year. Studies have shown that since the introduction of tobacco control measures, national smoking prevalence has declined in South Africa (SA). To determine the prevalence of tobacco smoking over a 7-year period in five impoverished neighbourhoods in Johannesburg, SA. Data were collected through the annual administration of a prestructured questionnaire to one adult respondent in preselected dwellings from 2006 to 2012. Information was collected on socioeconomic status, smoking practices and health status. Over the 7-year period of the analysis, smoking levels remained unchanged. The proportion of households with one or more smokers varied significantly across the five study neighbourhoods. Approximately 20% of households in Hillbrow and as many as 77% in Riverlea had a member who smoked. Despite a national downward trend in smoking levels, tobacco use remains high and persistent in certain vulnerable communities, requiring scaled-up action to reduce the risk of a range of tobacco-related diseases.


Renal phosphate handling and inherited disorders of phosphate reabsorption: an update.
Renal phosphate handling critically determines plasma phosphate and whole body phosphate levels. Filtered phosphate is mostly reabsorbed by Na+-dependent phosphate transporters located in the brush border membrane of the proximal tubule: NaPi-IIa (SLC34A1), NaPi-IIc (SLC34A3), and Pit-2 (SLC20A2). Here we review new evidence for the role and relevance of these transporters in inherited disorders of renal phosphate handling. The importance of NaPi-IIa and NaPi-IIc for renal phosphate reabsorption and mineral homeostasis has been highlighted by the identification of mutations in these transporters in a subset of patients with infantile idiopathic hypercalcemia and patients with hereditary hypophosphatemic rickets with hypercalciuria. Both diseases are characterized by disturbed calcium homeostasis secondary to elevated 1,25-(OH)2 vitamin D3 as a consequence of hypophosphatemia. In vitro analysis of mutated NaPi-IIa or NaPi-IIc transporters suggests defective trafficking underlying disease in most cases. Monoallelic pathogenic mutations in both SLC34A1 and SLC34A3 appear to be very frequent in the general population and have been associated with kidney stones. Consistent with these findings, results from genome-wide association studies indicate that variants in SLC34A1 are associated with a higher risk to develop kidney stones and chronic kidney disease, but underlying mechanisms have not been addressed to date.


Active-Site Engineering of ω-Transaminase for Production of Unnatural Amino Acids Carrying a Side Chain Bulkier than an Ethyl Substituent.
ω-Transaminase (ω-TA) is a promising enzyme for use in the production of unnatural amino acids from keto acids using cheap amino donors such as isopropylamine. The small substrate-binding pocket of most ω-TAs permits entry of substituents no larger than an ethyl group, which presents a significant challenge to the preparation of structurally diverse unnatural amino acids. Here we report on the engineering of an (S)-selective ω-TA from Ochrobactrum anthropi (OATA) to reduce the steric constraint and thereby allow the small pocket to readily accept bulky substituents. On the basis of a docking model in which L-alanine was used as a ligand, nine active-site residues were selected for alanine scanning mutagenesis. Among the resulting variants, an L57A variant showed dramatic activity improvements in activity for α-keto acids and α-amino acids carrying substituents whose bulk is up to that of an n-butyl substituent (e.g., 48- and 56-fold increases in activity for 2-oxopentanoic acid and L-norvaline, respectively). An L57G mutation also relieved the steric constraint but did so much less than the L57A mutation did. In contrast, an L57V substitution failed to induce the improvements in activity for bulky substrates. Molecular modeling suggested that the alanine substitution of L57, located in a large pocket, induces an altered binding orientation of an α-carboxyl group and thereby provides more room to the small pocket. The synthetic utility of the L57A variant was demonstrated by carrying out the production of optically pure L- and D-norvaline (i.e., enantiomeric excess [ee]>99%) by asymmetric amination of 2-oxopantanoic acid and kinetic resolution of racemic norvaline, respectively.


Checkpoint blockade immunotherapy reshapes the high-dimensional phenotypic heterogeneity of murine intratumoural neoantigen-specific CD8+ T cells.
The analysis of neoantigen-specific CD8+ T cells in tumour-bearing individuals is challenging due to the small pool of tumour antigen-specific T cells. Here we show that mass cytometry with multiplex combinatorial tetramer staining can identify and characterize neoantigen-specific CD8+ T cells in mice bearing T3 methylcholanthrene-induced sarcomas that are susceptible to checkpoint blockade immunotherapy. Among 81 candidate antigens tested, we identify T cells restricted to two known neoantigens simultaneously in tumours, spleens and lymph nodes in tumour-bearing mice. High-dimensional phenotypic profiling reveals that antigen-specific, tumour-infiltrating T cells are highly heterogeneous. We further show that neoantigen-specific T cells display a different phenotypic profile in mice treated with anti-CTLA-4 or anti-PD-1 immunotherapy, whereas their peripheral counterparts are not affected by the treatments. Our results provide insights into the nature of neoantigen-specific T cells and the effects of checkpoint blockade immunotherapy.Immune checkpoint blockade (ICB) therapies can unleash anti-tumour T-cell responses. Here the authors show, by integrating MHC tetramer multiplexing, mass cytometry and high-dimensional analyses, that neoantigen-specific, tumour-infiltrating T cells are highly heterogeneous and are subjected to ICB modulations.


Synthesis of thioether andrographolide derivatives and their inhibitory effect against cancer cells.
A series of novel thioether andrographolide derivatives were synthesized by incorporating various aromatic (or heteroaromatic) substituents into C-12 or 14-OH. A total of 38 andrographolide derivatives were prepared and evaluated for their in vitro inhibitory activity against cancer cells. All the derivatives exhibited better activity against prostate cancer cells (PC-3) than the parent compound. Among these, compounds 6a, 8, 9, 17, 19, 31, and 32 demonstrated good activity. These compounds were further evaluated for their anticancer activities against other cancer cell lines including MCF-7, MDA-MB-231, and A549. Compounds 31 and 32 showed excellent activity against MCF-7 with an IC50 value of 0.7 and 0.6 μM, respectively. The absolute configuration of 15a was determined via single-crystal X-ray diffraction. The activity of 6a (12S), which was the precursor of 15a, was better than that of the diastereoisomer 6b (12R). Moreover, the preliminary structure-activity relationship has been summarized. The results obtained herein are very important for further optimization of andrographolide.


A constrained approach to multiscale stochastic simulation of chemically reacting systems.
Stochastic simulation of coupled chemical reactions is often computationally intensive, especially if a chemical system contains reactions occurring on different time scales. In this paper, we introduce a multiscale methodology suitable to address this problem, assuming that the evolution of the slow species in the system is well approximated by a Langevin process. It is based on the conditional stochastic simulation algorithm (CSSA) which samples from the conditional distribution of the suitably defined fast variables, given values for the slow variables. In the constrained multiscale algorithm (CMA) a single realization of the CSSA is then used for each value of the slow variable to approximate the effective drift and diffusion terms, in a similar manner to the constrained mean-force computations in other applications such as molecular dynamics. We then show how using the ensuing Fokker-Planck equation approximation, we can in turn approximate average switching times in stochastic chemical systems.


Transcriptome analysis reveals novel players in the cranial neural crest gene regulatory network.
The neural crest is an embryonic stem cell population that gives rise to a multitude of derivatives. In particular, the cranial neural crest (CNC) is unique in its ability to contribute to both facial skeleton and peripheral ganglia. To gain further insight into the molecular underpinnings that distinguish the CNC from other embryonic tissues, we have utilized a CNC-specific enhancer as a tool to isolate a pure, region-specific NC subpopulation for transcriptional profiling. The resulting data set reveals previously unknown transcription factors and signaling pathways that may influence the CNC's ability to migrate and/or differentiate into unique derivatives. To elaborate on the CNC gene regulatory network, we evaluated the effects of knocking down known neural plate border genes and early neural crest specifier genes on selected neural crest-enriched transcripts. The results suggest that ETS1 and SOX9 may act as pan-neural crest regulators of the migratory CNC. Taken together, our analysis provides unprecedented characterization of the migratory CNC transcriptome and identifies new links in the gene regulatory network responsible for development of this critical cell population.


Genome mapping and characterization of the Anopheles gambiae heterochromatin.
Heterochromatin plays an important role in chromosome function and gene regulation. Despite the availability of polytene chromosomes and genome sequence, the heterochromatin of the major malaria vector Anopheles gambiae has not been mapped and characterized. To determine the extent of heterochromatin within the An. gambiae genome, genes were physically mapped to the euchromatin-heterochromatin transition zone of polytene chromosomes. The study found that a minimum of 232 genes reside in 16.6 Mb of mapped heterochromatin. Gene ontology analysis revealed that heterochromatin is enriched in genes with DNA-binding and regulatory activities. Immunostaining of the An. gambiae chromosomes with antibodies against Drosophila melanogaster heterochromatin protein 1 (HP1) and the nuclear envelope protein lamin Dm0 identified the major invariable sites of the proteins' localization in all regions of pericentric heterochromatin, diffuse intercalary heterochromatin, and euchromatic region 9C of the 2R arm, but not in the compact intercalary heterochromatin. To better understand the molecular differences among chromatin types, novel Bayesian statistical models were developed to analyze genome features. The study found that heterochromatin and euchromatin differ in gene density and the coverage of retroelements and segmental duplications. The pericentric heterochromatin had the highest coverage of retroelements and tandem repeats, while intercalary heterochromatin was enriched with segmental duplications. We also provide evidence that the diffuse intercalary heterochromatin has a higher coverage of DNA transposable elements, minisatellites, and satellites than does the compact intercalary heterochromatin. The investigation of 42-Mb assembly of unmapped genomic scaffolds showed that it has molecular characteristics similar to cytologically mapped heterochromatin. Our results demonstrate that Anopheles polytene chromosomes and whole-genome shotgun assembly render the mapping and characterization of a significant part of heterochromatic scaffolds a possibility. These results reveal the strong association between characteristics of the genome features and morphological types of chromatin. Initial analysis of the An. gambiae heterochromatin provides a framework for its functional characterization and comparative genomic analyses with other organisms.


Role clarification processes for better integration of nurse practitioners into primary healthcare teams: a multiple-case study.
Role clarity is a crucial issue for effective interprofessional collaboration. Poorly defined roles can become a source of conflict in clinical teams and reduce the effectiveness of care and services delivered to the population. Our objective in this paper is to outline processes for clarifying professional roles when a new role is introduced into clinical teams, that of the primary healthcare nurse practitioner (PHCNP). To support our empirical analysis we used the Canadian National Interprofessional Competency Framework, which defines the essential components for role clarification among professionals. A qualitative multiple-case study was conducted on six cases in which the PHCNP role was introduced into primary care teams. Data collection included 34 semistructured interviews with key informants involved in the implementation of the PHCNP role. Our results revealed that the best performing primary care teams were those that used a variety of organizational and individual strategies to carry out role clarification processes. From this study, we conclude that role clarification is both an organizational process to be developed and a competency that each member of the primary care team must mobilize to ensure effective interprofessional collaboration.


Iridium-based complexes for water oxidation.
Organometallic Ir precatalysts have been found to yield homogeneous Ir-based water-oxidation catalysts (WOCs) with very high activity. The Cp*Ir catalyst series can operate under a variety of regimes: it can either act as a homogeneous or a heterogeneous catalyst; it can be driven by chemical, photochemical, or electrochemical methods; and the molecular catalyst can either act in solution or supported as a molecular unit on a variety of solid oxides. In addition to optimizing the various reaction conditions, work has continued to elucidate the catalyst activation mechanism and identify water-oxidation intermediates. This Perspective will describe the development of the Cp*Ir series, their many forms as WOCs, and their ongoing characterization.


[Sequelae of collective violence: victims' voices in the ISAVIC study].
Despite the significant influence of collective violence on the health status of its victims, there are still many gaps in our understanding of the nature of the functional limitations this violence produces and its impact on victims' wellbeing and quality of life. The ISAVIC study was carried out in the autonomous region of the Basque Country from 2005-2008 to estimate the effects of collective violence on health. The assessment included victims' perceptions of these sequelae and their impact on health. A purposive sample of 36 primary victims was selected through contact networks and mediators. The victims' perceptions were collected through semi-structured in-depth interviews, which were later analyzed according to the study's conceptual framework. The victims' testimony suggests that they were profoundly traumatized by the collective violence experienced, which was often not yet completely overcome and significantly impaired their quality of life. The main functional, physical, emotional and social limitations described by the victims were identified. The qualitative results of this study are coherent with those obtained through the parametric phase of the ISAVIC study and provide a more complete overview of the nature of the sequelae of collective violence and its impact on quality of life. These results should be verified in larger studies and the influence of the social context on the relationship between collective violence and health should be analyzed in greater depth.


Changes in the rates of weight and waist circumference gain in Australian adults over time: a longitudinal cohort study.
To assess in a single cohort whether annual weight and waist circumference (WC) change has varied over time. Longitudinal cohort study with three surveys (1) 1999/2000; (2) 2004/2005 and (3) 2011/2012. Generalised linear mixed models with random effects were used to compare annualised weight and WC change between surveys 1 and 2 (period 1) with that between surveys 2 and 3 (period 2). Models were adjusted for age to analyse changes with time rather than age. Models were additionally adjusted for sex, education status, area-level socioeconomic disadvantage, ethnicity, body mass index, diabetes status and smoking status. The Australian Diabetes, Obesity and Lifestyle study (AusDiab)-a population-based, stratified-cluster survey of 11247 adults aged ≥25 years. 3351 Australian adults who attended each of three surveys and had complete measures of weight, WC and covariates. Weight and WC were measured at each survey. Change in weight and WC was annualised for comparison between the two periods. Mean weight and WC increased in both periods (0.34 kg/year, 0.43 cm/year period 1; 0.13 kg/year, 0.46 cm/year period 2). Annualised weight gain in period 2 was 0.11 kg/year (95% CI 0.06 to 0.15) less than period 1. Lesser annual weight gain between the two periods was not seen for those with greatest area-level socioeconomic disadvantage, or in men over the age of 55. In contrast, the annualised WC increase in period 2 was greater than period 1 (0.07 cm/year, 95% CI 0.01 to 0.12). The increase was greatest in men aged 55+ years and those with a greater area-level socioeconomic disadvantage. Between 2004/2005 and 2011/2012, Australian adults in a national study continued to gain weight, but more slowly than 1999/2000-2004/2005. While weight gain may be slowing, this was not observed for older men or those in more disadvantaged groups, and the same cannot be said for WC.


A Quantitative Method for Acesulfame K Using the Taste Sensor.
We have developed a method to quantify the sweetness of negatively charged high-potency sweeteners coexisting with other taste substances. This kind of sweetness sensor uses lipid polymer membranes as the taste-sensing part. Two types of outputs have been defined in the measurement of the taste sensor: one is the relative value and the other is the CPA (the change in membrane potential caused by adsorption) value. The CPA value shows a good selectivity for high-potency sweeteners. On the other hand, the relative value is several times higher than the CPA value, but the relative value is influenced by salty substances. In order to obtain both high sensitivity and selectivity, we established a model for predicting the concentration of sweeteners with a nonlinear regression analysis method using the relative values of both the sweetness sensor and the saltiness sensor. The analysis results showed good correlations with the estimated concentration of acesulfame potassium coexisting with salty substances, as represented by R2 = 0.99. This model can correspond well to the prediction of acesulfame K in a concentration of 0.2-0.7 mM, which is commonly used in food and beverages. The results obtained in this paper suggest that this method is useful for the evaluation of acesulfame K using the taste sensors.


2016 New Horizons Lecture: Beyond Imaging-Radiology of Tomorrow.
This article is based on the New Horizons lecture delivered at the 2016 Radiological Society of North America Annual Meeting. It addresses looming changes for radiology, many of which stem from the disruptive effects of the Fourth Industrial Revolution. This is an emerging era of unprecedented rapid innovation marked by the integration of diverse disciplines and technologies, including data science, machine learning, and artificial intelligence-technologies that narrow the gap between man and machine. Technologic advances and the convergence of life sciences, physical sciences, and bioengineering are creating extraordinary opportunities in diagnostic radiology, image-guided therapy, targeted radionuclide therapy, and radiology informatics, including radiologic image analysis. This article uses the example of oncology to make the case that, if members in the field of radiology continue to be innovative and continuously reinvent themselves, radiology can play an ever-increasing role in both precision medicine and value-driven health care. © RSNA, 2018.


Promoter influence on baculovirus-mediated gene expression in permissive and nonpermissive insect cell lines.
The activities of viral and insect promoters were examined in a range of insect cell lines permissive and nonpermissive for the replication of the baculovirus Autographa californica nuclear polyhedrosis virus. Recombinant baculoviruses were constructed to place the bacterial chloramphenicol acetyltransferase gene under the control of promoters strongly active in the early, late, or very late stages of virus replication. In fully permissive cells, expression from a very late promoter was 2- to 3-fold higher than expression from a late promoter and 10- to 20-fold higher than expression from an early promoter or from a virus-borne insect promoter. In cell lines that do not support the efficient production of viral progeny, late-promoter-driven expression was similar to or surpassed very late promoter-driven expression. In nonpermissive insect cell lines, expression driven by an insect promoter derived from Drosophila melanogaster was higher than expression from the three viral promoters and was especially high in the Drosophila cell line tested. Surprisingly, late-promoter-driven expression, which is dependent on DNA replication, was higher than early-promoter-driven expression in three of four nonpermissive lines. In contrast, very late promoter-driven expression was quite limited in nonpermissive cell lines. The results indicate that the promoter used to drive foreign-gene expression strongly influences the range of insect cells which can efficiently support the production of the foreign protein during infection with recombinant baculoviruses.


Incidence and routes of transmission of hepatitis B virus in England and Wales, 1995-2000: implications for immunisation policy.
The incidence of hepatitis B virus (HBV) infection in the UK is low. Since the infection can have serious sequelae, there is a continuing need to examine its epidemiology so as to inform control measures. We aimed to describe the current HBV incidence and patterns of transmission in the UK, to estimate the rate of new carrier infections, and to discuss implications for the control of HBV through immunisation. We analysed routine England and Wales laboratory surveillance data of acute HBV infection (1995-2000) and data on migration and global HBsAg prevalence. The estimated annual incidence of HBV infection in England and Wales was 7.4 per 100,000. Injecting drug use was the most frequently reported route of transmission. The number of cases attributed to heterosexual contact was fairly stable, whereas the number of cases in men having sex with men decreased. These observations continue trends reported for the early 1990s. Transmission during childhood was rarely reported, but was more frequent among South Asians. The incidence in South Asians is relatively high, and their main risk factors are medical treatment overseas and heterosexual contact. For about a third of cases of acute HBV infection no route of transmission is reported, but analysis of secular trends and age distribution suggest that many of these may be related to injecting drug use. Endemic transmission gives rise to only a small proportion of all new chronic infections, with the vast majority arising from immigration of established HBV carriers. The incidence of acute HBV infection in England and Wales has remained low, with a similar pattern of reported routes of transmission compared to the early 1990s. The UK prevalence of HBV infection is dependant on global rather than national immunisation policy. Endemic transmission may be reduced by improving immunisation coverage among injecting drug users, which is expected to also reduce the number of cases without a risk factor reported. In addition, immunisation options that better suit the needs of ethnic minorities need to be explored.


Defining risk groups to yellow fever vaccine-associated viscerotropic disease in the absence of denominator data.
Several risk groups are known for the rare but serious, frequently fatal, viscerotropic reactions following live yellow fever virus vaccine (YEL-AVD). Establishing additional risk groups is hampered by ignorance of the numbers of vaccinees in factor-specific risk groups thus preventing their use as denominators in odds ratios (ORs). Here, we use an equation to calculate ORs using the prevalence of the factor-specific risk group in the population who remain well. The 95% confidence limits and P values can also be calculated. Moreover, if the estimate of the prevalence is imprecise, discrimination analysis can indicate the prevalence at which the confidence interval results in an OR of ∼1 revealing if the prevalence might be higher without yielding a non-significant result. These methods confirm some potential risk groups for YEL-AVD and cast doubt on another. They should prove useful in situations in which factor-specific risk group denominator data are not available.


Predictors of women's perceptions of the childbirth experience.
To determine the factors that predict women's perceptions of the childbirth experience and to examine whether these vary with the type of birth a woman experiences. Prospective cohort study. The postpartum units of two eastern Canadian hospitals. Six hundred fifty two women and their newborns. Data were collected in hospital at 12 to 48 hours postpartum using self-report questionnaires and chart review. Perception of the childbirth experience was measured for women having a vaginal and emergency cesarean birth using the Questionnaire Measuring Attitudes About Labor and Delivery and planned cesarean birth using the Modified Questionnaire Measuring Attitudes About Labor and Delivery. Of the 20 predictors of women's childbirth perceptions, the strongest were type of birth; degree of awareness, relaxation, and control; helpfulness of partner support; and being together with the infant following birth. Of the predictors of a quality birth experience, most were amenable to nursing interventions: enhancement of patient awareness, relaxation, and control; promotion of partner support; and provision of immediate opportunities for women to be with their babies.


doRiNA: a database of RNA interactions in post-transcriptional regulation.
In animals, RNA binding proteins (RBPs) and microRNAs (miRNAs) post-transcriptionally regulate the expression of virtually all genes by binding to RNA. Recent advances in experimental and computational methods facilitate transcriptome-wide mapping of these interactions. It is thought that the combinatorial action of RBPs and miRNAs on target mRNAs form a post-transcriptional regulatory code. We provide a database that supports the quest for deciphering this regulatory code. Within doRiNA, we are systematically curating, storing and integrating binding site data for RBPs and miRNAs. Users are free to take a target (mRNA) or regulator (RBP and/or miRNA) centric view on the data. We have implemented a database framework with short query response times for complex searches (e.g. asking for all targets of a particular combination of regulators). All search results can be browsed, inspected and analyzed in conjunction with a huge selection of other genome-wide data, because our database is directly linked to a local copy of the UCSC genome browser. At the time of writing, doRiNA encompasses RBP data for the human, mouse and worm genomes. For computational miRNA target site predictions, we provide an update of PicTar predictions.


Imported case of acute respiratory tract infection associated with a member of species nelson bay orthoreovirus.
A Japanese man suffered from acute respiratory tract infection after returning to Japan from Bali, Indonesia in 2007. Miyazaki-Bali/2007, a strain of the species of Nelson Bay orthoreovirus, was isolated from the patient's throat swab using Vero cells, in which syncytium formation was observed. This is the sixth report describing a patient with respiratory tract infection caused by an orthoreovirus classified to the species of Nelson Bay orthoreovirus. Given the possibility that all of the patients were infected in Malaysia and Indonesia, prospective surveillance on orthoreovirus infections should be carried out in Southeast Asia. Furthermore, contact surveillance study suggests that the risk of human-to-human infection of the species of Nelson Bay orthoreovirus would seem to be low.


Miswiring of Frontostriatal Projections in Schizophrenia.
We investigated brain wiring in chronic schizophrenia and healthy controls in frontostriatal circuits using diffusion magnetic resonance imaging tractography in a novel way. We extracted diffusion streamlines in 27 chronic schizophrenia and 26 healthy controls connecting 4 frontal subregions to the striatum. We labeled the projection zone striatal surface voxels into 2 subtypes: dominant-input from a single cortical subregion, and, functionally integrative, with mixed-input from diverse cortical subregions. We showed: 1) a group difference for total striatal surface voxel number (P = .045) driven by fewer mixed-input voxels in the left (P = .007), but not right, hemisphere; 2) a group by hemisphere interaction for the ratio quotient between voxel subtypes (P = .04) with a left (P = .006), but not right, hemisphere increase in schizophrenia, also reflecting fewer mixed-input voxels; and 3) fewer mixed-input voxel counts in schizophrenia (P = .045) driven by differences in left hemisphere limbic (P = .007) and associative (P = .01), but not sensorimotor, striatum. These results demonstrate a less integrative pattern of frontostriatal structural connectivity in chronic schizophrenia. A diminished integrative pattern yields a less complex input pattern to the striatum from the cortex with less circuit integration at the level of the striatum. Further, as brain wiring occurs during early development, aberrant brain wiring could serve as a developmental biomarker for schizophrenia.


Cardiac magnetic resonance imaging: a new tool to identify cardioaortic sources in ischaemic stroke.
Stroke of undetermined aetiology or 'cryptogenic' stroke accounts for 30-40% of ischaemic strokes despite extensive diagnostic evaluation. The role and yield of cardiac imaging is controversial. Cardiac MRI (CMR) has been used for cardiac disorders, but its use in cryptogenic stroke is not well established. We reviewed the literature (randomised trials, exploratory comparative studies and case series) on the use of CMR in the diagnostic evaluation of patients with ischaemic stroke. The literature on the use of CMR in the diagnostic evaluation of ischaemic stroke is sparse. However, studies have demonstrated a potential role for CMR in the diagnostic evaluation of patients with cryptogenic stroke to identify potential aetiologies such as cardiac thrombi, cardiac tumours, aortic arch disease and other rare cardiac anomalies. CMR can also provide data on certain functional and structural parameters of the left atrium and the left atrial appendage which have been shown to be associated with ischaemic stroke risk. CMR is a non-invasive modality that can help identify potential mechanisms in cryptogenic stroke and patients who may be targeted for enrolment into clinical trials comparing anticoagulation to antiplatelet therapy in secondary stroke prevention. Prospective studies are needed to compare the value of CMR as compared to transthoracic and transesophageal echocardiography in the diagnostic evaluation of cryptogenic stroke.


A biosensor for MAPK-dependent Lin28 signaling.
Intracellular levels of the RNA-binding protein and pluripotency factor, Lin28a, are tightly controlled to govern cellular and organismal growth. Lin28a is extensively regulated at the posttranscriptional level, and can undergo mitogen-activated protein kinase (MAPK)-mediated elevation from low basal levels in differentiated cells by phosphorylation-dependent stabilizing interaction with the RNA-silencing factor HIV TAR RNA-binding protein (TRBP). However, molecular and spatiotemporal details of this critical control mechanism remained unknown. In this work, we dissect the interacting regions of Lin28a and TRBP proteins and develop biosensors to visualize this interaction. We identify truncated domains of Lin28a and of TRBP that are sufficient to support coassociation and mutual elevation of protein levels, and a requirement for MAPK-dependent phosphorylation of TRBP at putative Erk-target serine 152, as well as Lin28a serine 200 phosphorylation, in mediating the increase of Lin28a protein by TRBP. The phosphorylation-dependent association of Lin28a and TRBP truncated constructs is leveraged to develop fluorescence resonance energy transfer (FRET)-based sensors for dynamic monitoring of Lin28a and TRBP interaction. We demonstrate the response of bimolecular and unimolecular FRET sensors to growth factor stimulation in living cells, with coimaging of Erk activation to achieve further understanding of the role of MAPK signaling in Lin28a regulation.


Public health impact of adverse bone effects of oral corticosteroids.
The objective of this study was to estimate the number of fractures attributed to oral corticosteroid use. Information was obtained from the General Practice Research Database which contains medical records of general practitioners in the UK. The total number of corticosteroid-related fractures during a course of treatment was estimated using the formula for attributable risk among the exposed. A total of 244 235 patients was prescribed an oral corticosteroid. The rate of hip fractures increased exponentially with age in both males and females. The excess number of hip fracture cases among females aged 85 years or older using 7.5 mg prednisolone per day or more was 1.4 cases per 100 patients per year. About 47% of all hip and 72% of all vertebral fractures that occurred can be attributed to oral corticosteroid use. Among 10 000 female users of higher doses, 99.7 nonvertebral, 31.6 hip and 45.8 vertebral fractures can be attributed to use of oral corticosteroids. The targeting of high-risk patients will be important for implementing preventative strategies in a cost-effective manner.


Impact of a resistance gene against a fungal pathogen on the plant host residue microbiome: The case of the Leptosphaeria maculans-Brassica napus pathosystem.
Oilseed rape residues are a crucial determinant of stem canker epidemiology as they support the sexual reproduction of the fungal pathogen Leptosphaeria maculans. The aim of this study was to characterize the impact of a resistance gene against L. maculans infection on residue microbial communities and to identify microorganisms interacting with this pathogen during residue degradation. We used near-isogenic lines to obtain healthy and infected host plants. The microbiome associated with the two types of plant residues was characterized by metabarcoding. A combination of linear discriminant analysis and ecological network analysis was used to compare the microbial communities and to identify microorganisms interacting with L. maculans. Fungal community structure differed between the two lines at harvest, but not subsequently, suggesting that the presence/absence of the resistance gene influences the microbiome at the base of the stem whilst the plant is alive, but that this does not necessarily lead to differential colonization of the residues by fungi. Direct interactions with other members of the community involved many fungal and bacterial amplicon sequence variants (ASVs). L. maculans appeared to play a minor role in networks, whereas one ASV affiliated to Plenodomus biglobosus (synonym Leptosphaeria biglobosa) from the Leptosphaeria species complex may be considered a keystone taxon in the networks at harvest. This approach could be used to identify and promote microorganisms with beneficial effects against residue-borne pathogens and, more broadly, to decipher the complex interactions between multispecies pathosystems and other microbial components in crop residues.


Estimation and Optimization of Composite Outcomes.
There is tremendous interest in precision medicine as a means to improve patient outcomes by tailoring treatment to individual characteristics. An individualized treatment rule formalizes precision medicine as a map from patient information to a recommended treatment. A treatment rule is defined to be optimal if it maximizes the mean of a scalar outcome in a population of interest, e.g., symptom reduction. However, clinical and intervention scientists often seek to balance multiple and possibly competing outcomes, e.g., symptom reduction and the risk of an adverse event. One approach to precision medicine in this setting is to elicit a composite outcome which balances all competing outcomes; unfortunately, eliciting a composite outcome directly from patients is difficult without a high-quality instrument, and an expert-derived composite outcome may not account for heterogeneity in patient preferences. We propose a new paradigm for the study of precision medicine using observational data that relies solely on the assumption that clinicians are approximately (i.e., imperfectly) making decisions to maximize individual patient utility. Estimated composite outcomes are subsequently used to construct an estimator of an individualized treatment rule which maximizes the mean of patient-specific composite outcomes. The estimated composite outcomes and estimated optimal individualized treatment rule provide new insights into patient preference heterogeneity, clinician behavior, and the value of precision medicine in a given domain. We derive inference procedures for the proposed estimators under mild conditions and demonstrate their finite sample performance through a suite of simulation experiments and an illustrative application to data from a study of bipolar depression.


Perturbation of Hydrogen-Bonding Networks over Supported Lipid Bilayers by Poly(allylamine hydrochloride).
Water is vital to many biochemical processes and is necessary for driving fundamental interactions of cell membranes with their external environments, yet it is difficult to probe the membrane/water interface directly and without the use of external labels. Here, we employ vibrational sum frequency generation spectroscopy to understand the role of interfacial water molecules above bilayers formed from zwitterionic (phosphatidylcholine) and anionic (phosphatidylglycerol, PG, and phosphatidylserine, PS) lipids as they are exposed to the common polycation poly(allylamine hydrochloride) (PAH) in 100 mM NaCl. We show that as the concentration of PAH is increased, the interfacial water molecules are irreversibly displaced and find that it requires 10 times more PAH to displace interfacial water molecules from membranes formed from purely zwitterionic lipids when compared to membranes that contain the anionic PG and PS lipids. This outcome is likely due to the difference in (1) the energy with which water molecules are bound to the lipid headgroups, (2) the number of water molecules bound to the headgroups, which is related to the headgroup area, and (3) the electrostatic interactions between the PAH molecules and the negatively charged lipids that are favored when compared to the zwitterionic lipid headgroups. The findings presented here contribute to establishing causal relationships in nanotoxicology and to understanding, controlling, and predicting the initial steps that lead to the lysis of cells exposed to membrane-disrupting polycations or to transfection.


Effects of corticosteroids on COPD lung macrophage phenotype and function.
The numbers of macrophages are increased in the lungs of chronic obstructive pulmonary disease (COPD) patients. COPD lung macrophages have reduced ability to phagocytose microbes and efferocytose apoptotic cells. Inhaled corticosteroids (ICSs) are widely used anti-inflammatory drugs in COPD; however, their role beyond suppression of cytokine release has not been explored in COPD macrophages. We have examined the effects of corticosteroids on COPD lung macrophage phenotype and function. Lung macrophages from controls and COPD patients were treated with corticosteroids; effects on gene and protein expression of CD163, CD164, CD206, MERTK, CD64, CD80 and CD86 were studied. We also examined the effect of corticosteroids on the function of CD163, MERTK and cluster of differentiation 64 (CD64). Corticosteroid increased CD163, CD164, CD206 and MERTK expression and reduced CD64, CD80 and CD86 expression. We also observed an increase in the uptake of the haemoglobin-haptoglobin complex (CD163) from 59 up to 81% and an increase in efferocytosis of apoptotic neutrophils (MERTK) from 15 up to 28% following corticosteroid treatment. We observed no effect on bacterial phagocytosis. Corticosteroids alter the phenotype and function of COPD lung macrophages. Our findings suggest mechanisms by which corticosteroids exert therapeutic benefit in COPD, reducing iron available for bacterial growth and enhancing efferocytosis.


Molecular Minimal Residual Disease in Acute Myeloid Leukemia.
Patients with acute myeloid leukemia (AML) often reach complete remission, but relapse rates remain high. Next-generation sequencing enables the detection of molecular minimal residual disease in virtually every patient, but its clinical value for the prediction of relapse has yet to be established. We conducted a study involving patients 18 to 65 years of age who had newly diagnosed AML. Targeted next-generation sequencing was carried out at diagnosis and after induction therapy (during complete remission). End points were 4-year rates of relapse, relapse-free survival, and overall survival. At least one mutation was detected in 430 out of 482 patients (89.2%). Mutations persisted in 51.4% of those patients during complete remission and were present at various allele frequencies (range, 0.02 to 47%). The detection of persistent DTA mutations (i.e., mutations in DNMT3A, TET2, and ASXL1), which are often present in persons with age-related clonal hematopoiesis, was not correlated with an increased relapse rate. After the exclusion of persistent DTA mutations, the detection of molecular minimal residual disease was associated with a significantly higher relapse rate than no detection (55.4% vs. 31.9%; hazard ratio, 2.14; P<0.001), as well as with lower rates of relapse-free survival (36.6% vs. 58.1%; hazard ratio for relapse or death, 1.92; P<0.001) and overall survival (41.9% vs. 66.1%; hazard ratio for death, 2.06; P<0.001). Multivariate analysis confirmed that the persistence of non-DTA mutations during complete remission conferred significant independent prognostic value with respect to the rates of relapse (hazard ratio, 1.89; P<0.001), relapse-free survival (hazard ratio for relapse or death, 1.64; P=0.001), and overall survival (hazard ratio for death, 1.64; P=0.003). A comparison of sequencing with flow cytometry for the detection of residual disease showed that sequencing had significant additive prognostic value. Among patients with AML, the detection of molecular minimal residual disease during complete remission had significant independent prognostic value with respect to relapse and survival rates, but the detection of persistent mutations that are associated with clonal hematopoiesis did not have such prognostic value within a 4-year time frame. (Funded by the Queen Wilhelmina Fund Foundation of the Dutch Cancer Society and others.).


Supplemental cracked corn for steers fed fresh alfalfa: II. Protein and amino acid digestion.
The effects of different levels of cracked corn on N intake, ruminal bacterial CP synthesis, and duodenal flows and small intestinal digestion of amino acids (AA) in steers fed fresh alfalfa indoors were determined. Angus steers (n = 6; average BW 338 +/- 19 kg) cannulated in the rumen, duodenum, and ileum were fed each of five diets over five periods in a Latin square design with an extra animal. Steers consumed 1) alfalfa (20.4% CP, 41.6% NDF) ad libitum (AALF); 2), 3), and 4) AALF supplemented (S) with three levels of corn (.4, .8, or 1.2% of BW, respectively), or 5) alfalfa restricted (RALF) to the average forage intake of S steers. Average N intake and duodenal flow of nonammonia N (NAN) were greater (P < .01) in S than in RALF steers. Greater duodenal flows of NAN in S compared with RALF were due to a trend toward higher (P = .06) flows of both bacterial and dietary N. Levels of corn decreased (P < .01) linearly N intake and increased (P < .01) linearly duodenal flow of NAN owing to a numerical linear increase in nonbacterial N (P = .15) with no increase in bacterial N flow. Duodenal NAN flows as percentages of N intake increased (P < .01) linearly (69.3 to 91.0%) as corn increased. Ruminal NH3 N concentration, ruminal CP degradability, and the proportion of bacterial N in duodenal NAN were decreased (P < .01) linearly as corn increased. Efficiency of net microbial CP synthesis was not affected (P > .05) by treatment (average 42.6 and 30.9 g N/kg of OM apparently or truly digested in the rumen, respectively). Small intestinal disappearance of total N and individual AA, except for threonine and lysine, and small intestinal digestibility of N and individual AA, except for methionine, histidine, and proline, increased (P < .01) linearly with level of corn and were greater (P < .01) in S than in RALF steers. Supplementing corn to steers fed fresh alfalfa reduced ruminal N losses and CP degradability and increased the duodenal flow and the small intestinal disappearance and digestibility of total N and total, essential, and nonessential AA.


White matter tract and glial-associated changes in 5-hydroxymethylcytosine following chronic cerebral hypoperfusion.
White matter abnormalities due to age-related cerebrovascular alterations is a common pathological hallmark associated with functional impairment in the elderly which has been modeled in chronically hypoperfused mice. 5-Methylcytosine (5mC) and its oxidized derivative 5-hydroxymethylcytosine (5hmC) are DNA modifications that have been recently linked with age-related neurodegeneration and cerebrovascular pathology. Here we conducted a pilot investigation of whether chronic cerebral hypoperfusion might affect genomic distribution of these modifications and/ or a Ten-Eleven Translocation protein 2 (TET2) which catalyses hydroxymethylation in white and grey matter regions of this animal model. Immunohistochemical evaluation of sham and chronically hypoperfused mice a month after surgery revealed significant (p<0.05) increases in the proportion of 5hmC positive cells, Iba1 positive inflammatory microglia, and NG2 positive oligodendroglial progenitors in the hypoperfused corpus callosum. In the same white matter tract there was an absence of hypoperfusion-induced alterations in the proportion of 5mC, TET2 positive cells and CC1 positive mature oligodrendrocytes. Correlation analysis across animals within both treatment groups demonstrated a significant association of the elevated 5hmC levels with increases in the proportion of inflammatory microglia only (p=0.01) in the corpus callosum. In vitro studies revealed that 5hmC is lost during oligodendroglial maturation but not microglial activation. Additionally, TET1, TET2, and TET3 protein levels showed dynamic alterations during oligodendroglial development and following oxidative stress in vitro. Our study suggests that 5hmC exhibits white matter tract and cell type specific dynamics following chronic cerebral hypoperfusion in mice.


Chondrogenesis in chick limb bud mesodermal cells: reciprocal modulation by activin and inhibin.
Activin and inhibin are members of the transforming growth factor-beta superfamily sharing a common beta-subunit, but with opposite biological activities on the release of follicle-stimulating hormone from pituitary. Recent studies have demonstrated that activin is the long sought after mesoderm induction factor in amphibians. To determine if activin and inhibin affect the proliferation and differentiation of chick limb bud mesodermal cells during cartilage formation of the limb, recombinant human activin A and inhibin A were tested on stage 24 chick limb bud mesodermal cells. The results showed that activin A has an inhibitory effect on chondrogenic differentiation, as indicated by Alcian blue staining and decreased [35S]sulfate incorporation into proteoglycans. In addition, the expression of type II collagen mRNA, a specific phenotypic marker of cartilage, was greatly inhibited when cultures were exposed to activin A for 5 days at a dose of 10 ng/ml. In contrast, inhibin A stimulated cartilage formation, as indicated by increased expression of type II collagen mRNA and synthesis of proteoglycans. These results imply that activin A and inhibin A can modulate chondrogenesis during differentiation of limb bud cells in culture.


Contractile deactivation and uncoupling of crossbridges. Effects of 2,3-butanedione monoxime on mammalian myocardium.
We investigated the effects of 1 and 3 mM 2,3-butanedione monoxime (BDM, diacetyl monoxime) on excitation and contraction of cardiac muscle in several types of preparations at various levels of organization. We selected a concentration of BDM that was not expected to affect sarcolemmal calcium flux and action potential duration in cardiac tissue. Two indicators were used to record intracellular calcium. Aequorin, a bioluminescent calcium indicator, was used in studies with ferret papillary muscle preparations, and fura-2, a fluorescent calcium indicator, was used in studies with guinea pig cardiac myocytes. In both cases, addition of BDM resulted in a reduction of peak intracellular calcium released from the sarcoplasmic reticulum and a reduction of peak twitch force. The duration of the action potential of isolated myocytes was slightly abbreviated in the presence of BDM. In studies on the calcium current in the myocytes, addition of BDM was associated with reduced calcium current at any potential. Peak calcium current was reduced by 7.9 +/- 1% in the presence of BDM. In tetanized ferret papillary muscles, BDM reduced maximal calcium-activated force by 30 +/- 5% and increased the calcium ion concentration required for half-maximal force by 0.1 +/- 0.01 microM. The Hill coefficient was reduced from 5.00 +/- 0.11 to 3.40 +/- 0.20. Maximal shortening velocity of ferret papillary muscles was increased in the presence of BDM from 1.55 +/- 0.24 to 2.04 +/- 0.33 mm/sec. Ca2+ binding to troponin C in skinned fiber preparations from guinea pig, bovine, and canine hearts was unaffected by addition of up to 10 mM BDM. Our results indicate that BDM affects both calcium availability and responsiveness of the myofilaments to Ca2+. Uncoupling of contractile activation from excitation may also result from altered crossbridge kinetics.


Cortical activity measured by functional near infrared spectroscopy during a theory of mind task in subjects with schizophrenia, bipolar disorder and healthy controls.
Theory of Mind (ToM) deficits interfere in social cognitive functioning in schizophrenia (SCZ) and are increasingly recognized to do so in bipolar disorder (BD), however their clinical and neurobiological correlates remain unclear. This study represents the first direct comparison of subjects with SCZ (N = 26), BD (N = 26) and healthy controls (N = 33) in cortical activity during the Reading the Mind in the Eyes Task (RMET) using functional Near Infrared Spectroscopy (fNIRS) with the control condition (CC) involving gender identification via the same stimuli. The three groups were compared with a comprehensive ToM battery and assessed in terms of the relationship of ToM performance with clinical symptoms, insight and functioning. The controls scored higher than the SCZ and BD groups in ToM assessments, with SCZ group showing the worse performance in terms of meta-representation and empathy. The SCZ group ToM scores inversely correlated with negative symptom severity and positively correlated with insight; BD group ToM scores negatively correlated with subclinical mania symptoms and projected functioning. Cortical activity was higher during the ToM condition compared to the CC in the pre-motor and supplementary-motor cortices, middle and superior temporal gyri, and the primary somatosensory cortex. Group x Condition interaction was detected whereby activity was higher during the ToM condition among controls with no detected difference between SCZ and BD groups. The results suggest that ToM is represented similarly in cortical activity in SCZ and BD compared to healthy controls pointing to possible neurobiological convergence of SCZ and BD in underlying impairments of social cognition.


Tuning the Mechanical Response of Metal-Organic Frameworks by Defect Engineering.
The incorporation of defects into crystalline materials provides an important tool to fine-tune properties throughout various fields of materials science. We performed high-pressure powder X-ray diffraction experiments, varying pressures from ambient to 0.4 GPa in 0.025 GPa increments to probe the response of defective UiO-66 to hydrostatic pressure for the first time. We observe an onset of amorphization in defective UiO-66 samples around 0.2 GPa and decreasing bulk modulus as a function of defects. Intriguingly, the observed bulk moduli of defective UiO-66(Zr) samples do not correlate with defect concentration, highlighting the complexity of how defects are spatially incorporated into the framework. Our results demonstrate the large impact of point defects on the structural stability of metal-organic frameworks (MOFs) and pave the way for experiment-guided computational studies on defect engineered MOFs.


Conditional deletion of Hdac3 in osteoprogenitor cells attenuates diet-induced systemic metabolic dysfunction.
Obesity is a major health epidemic in the United States and a leading cause of preventable diseases including type 2 diabetes. A growing body of evidence indicates that the skeleton influences whole body metabolism and suggests a new avenue for developing novel therapeutic agents, but the underlying mechanisms are not well understood. Here, it is demonstrated that conditional deletion of an epigenetic regulator, Hdac3, in osteoblast progenitor cells abrogates high fat diet-induced insulin resistance and hepatic steatosis. These Hdac3-deficient mice have reduced bone formation and lower circulating levels of total and undercarboxylated osteocalcin, coupled with decreased bone resorption activity. They also maintain lower body fat and fasting glucose levels on normal and high fat chow diets. The mechanisms by which Hdac3 controls systemic energy homeostasis from within osteoblasts have not yet been fully realized, but the current study suggests that it does not involve elevated levels of circulating osteocalcin. Thus, Hdac3 is a new player in the emerging paradigm that the skeleton influences systemic energy metabolism.


Factors associated with BMI, weight perceptions and trying to lose weight in African-American smokers.
This study examined sociodemographic, behavioral and psychosocial factors associated with BMI, weight perceptions and trying to lose weight among African-American smokers (N=600, M=44.2 years, 70% female). Sixty-eight percent of the sample were overweight or obese (sample BMI M=28.0, SD=6.7). Three separate, simultaneous multivariable regression models were used to determine which factors were associated with BMI, weight perceptions and trying to lose weight. Poorer health, female gender and high-school education or higher were significantly associated with higher BMIs (p<0.05). Being female (OR=5.8, 95% CI=3.6-9.3) and having a higher BMI (OR=0.6, 95% CI=0.5-0.6) was associated with perception of overweight and smoking more cigarettes per day (OR=1.0, 95% CI=1.0-1.1), and perceiving oneself as overweight (OR=14.1, 95% CI=8.2-24.2) was associated with trying to lose weight. Participants somewhat underestimated their BMI in their weight perceptions. Those who perceived themselves as overweight were more likely to be trying to lose weight; therefore, increasing participant awareness of actual BMI status may lead to improved weight-control efforts in African-American smokers. Several expected associations with outcomes were not found, suggesting that BMI and weight constructs are not well-understood in this population.


Reduced Prefrontal Gyrification in Carriers of the Dopamine D4 Receptor 7-Repeat Allele With Attention Deficit/Hyperactivity Disorder: A Preliminary Report.
Objective: Structural and functional abnormalities have been noted in the prefrontal cortex of individuals with neurodevelopmental disorders such as attention deficit/hyperactivity disorder (ADHD). Cortical thickness and gyrification, both of which have been reported as abnormal in the prefrontal cortex in ADHD, are thought to be modulated by genetic influences during neural development. This study aimed to investigate the effects of a polymorphism of the dopamine DRD4 gene (the 7-repeat (7R) "risk" allele) on thickness and gyrification as distinct parameters of prefrontal cortical structure in children with ADHD. Method: Structural images and genetic samples were obtained from 49 children aged 9-15 years (25 with ADHD and 24 matched controls), and measures of cortical thickness and gyrification for inferior, middle, and superior frontal cortex were calculated. Results: A significant interaction between diagnosis and genotype on prefrontal gyrification was observed, largely driven by reduced inferior frontal gyrification in patients who carried the DRD4 7R allele. Furthermore, inferior frontal gyrification-but not thickness-related to everyday executive functioning in 7R allele carriers across groups. Conclusions: Prefrontal gyrification is reduced in children with ADHD who also carry the DRD4 7R allele, and it relates to critical functional skills in the executive domain in carriers of the risk allele. More broadly, these effects highlight the importance of considering precise neurodevelopmental mechanisms through which risk alleles influence cortical neurogenesis and migration.


Comparing Region of Interest versus Voxel-Wise Diffusion Tensor Imaging Analytic Methods in Mild and Moderate Traumatic Brain Injury: A Systematic Review and Meta-Analysis.
Diffusion tensor imaging is a magnetic resonance imaging technique that is uniquely capable of detecting microstructural tissue damage in mild and moderate traumatic brain injuries (TBIs). To date, it remains unknown if two common analytic techniques, region of interest (ROI) versus voxel-wise (VW) analyses, detect injury in similar locations. The purpose of this systematic review and meta-analysis was to directly compare the regions of abnormality elucidated by each method. Twenty-seven ROI and 11 VW studies met our inclusion criteria. Our ROI meta-analysis identified 11 regions, including the splenium of the corpus callosum, where fractional anisotropy (FA) was significantly decreased in TBI patients, compared with controls. Likewise, we identified higher mean diffusivity/apparent diffusivity constant in the genu, body, and splenium of the corpus callosum. Alternatively, our VW analysis identified one region of high FA in the right superior longitudinal fasciculus and seven regions of low FA, with the two largest located in the corpus callosum. High mean diffusivity and high radial diffusivity, both in the right inferior longitudinal fasciculus, also was revealed by our VW analysis. Moreover, we have shown that the magnitude of damage in the corpus callosum revealed by ROI analysis (z = -3.15) is greater than that demonstrated by VW analysis (z = -1.41). Overall, this study indicates that both ROI and VW analytic methods are sensitive to low FA in the corpus callosum; however, the ROI method has more power to detect the full extent of tissue abnormality in the corpus callosum. More research utilizing standardized methods and reporting is essential to fully characterize the extent to which ROI and VW analyses can concordantly detect other locations of pathology in mild and moderate TBI patients.


A Huaier polysaccharide reduced metastasis of human hepatocellular carcinoma SMMC-7721 cells via modulating AUF-1 signaling pathway.
TP-1 is a polysaccharide from one famous fungus Huaier. Treatment with TP-1 significantly inhibited the cell growth, adhesion, migration, and motility of SMMC-7721 cells in a dose-dependent manner. Real-time quantitative RT-PCR revealed a dose-dependent decrease in RNA-binding factor 1 (AUF-1) and astrocyte elevated gene-1 (AEG-1) messenger RNA (mRNA) levels in TP-1-treated SMMC-7721 cells, which is consistent with their protein expression detected by Western blotting. On the contrary, microRNA-122 (miR-122) expression increased in SMMC-7721 cells following TP-1 treatment. Moreover, TP-1 treatment at three doses apparently increased epithelial marker E-cadherin protein expression but decreased the mesenchymal marker N-cadherin protein level. In addition, the hematoxylin-eosin (H & E) staining showed that the TP-1 significantly inhibited the lung metastasis of liver cancer in mice orthotopic implanted with SMMC-7721 tumor tissue. Taken together, these findings proved the inhibitory effect of TP-1 on the growth and metastasis of SMMC-7721 cells, and TP-1 might be offered for future application as a powerful chemopreventive agent against hepatocellular carcinoma (HCC) metastasis.


Explaining, not just predicting, drives interest in personal genomics.
There is a widespread assumption that risk prediction is the major driver of customer interest in personal genomic testing (PGT). However, some customers may also be motivated by finding out whether their existing diseases have a genetic etiology. We evaluated the impact of an existing medical diagnosis on customer interest in condition-specific results from PGT. Using a prospective online survey of PGT customers, we measured customer interest prior to receiving PGT results for 11 health conditions, and examined the association between interest and personal medical history of these conditions using logistic regression. We analyzed data from 1,538 PGT customers, mean age 48.7 years, 61 % women, 90 % White, and 47 % college educated. The proportion of customers who were 'very interested' in condition-specific PGT varied considerably, from 28 % for ulcerative colitis to 68% for heart disease. After adjusting for demographic and personal characteristics including family history, having a diagnosis of the condition itself was significantly associated with interest in genetic testing for risk of that condition, with odds ratios ranging from 2.07 (95 % CI 1.28-3.37) for diabetes to 19.99 (95 % CI 4.57-87.35) for multiple sclerosis. PGT customers are particularly interested in genetic markers for their existing medical conditions, suggesting that the value of genetic testing is not only predictive, but also explanatory.


The Sustained Induction of c-MYC Drives Nab-Paclitaxel Resistance in Primary Pancreatic Ductal Carcinoma Cells.
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease with limited and, very often, ineffective medical and surgical therapeutic options. The treatment of patients with advanced unresectable PDAC is restricted to systemic chemotherapy, a therapeutic intervention to which most eventually develop resistance. Recently, nab-paclitaxel (n-PTX) has been added to the arsenal of first-line therapies, and the combination of gemcitabine and n-PTX has modestly prolonged median overall survival. However, patients almost invariably succumb to the disease, and little is known about the mechanisms underlying n-PTX resistance. Using the conditionally reprogrammed (CR) cell approach, we established and verified continuously growing cell cultures from treatment-naïve patients with PDAC. To study the mechanisms of primary drug resistance, nab-paclitaxel-resistant (n-PTX-R) cells were generated from primary cultures and drug resistance was verified in vivo, both in zebrafish and in athymic nude mouse xenograft models. Molecular analyses identified the sustained induction of c-MYC in the n-PTX-R cells. Depletion of c-MYC restored n-PTX sensitivity, as did treatment with either the MEK inhibitor, trametinib, or a small-molecule activator of protein phosphatase 2a. IMPLICATIONS: The strategies we have devised, including the patient-derived primary cells and the unique, drug-resistant isogenic cells, are rapid and easily applied in vitro and in vivo platforms to better understand the mechanisms of drug resistance and for defining effective therapeutic options on a patient by patient basis.


Quantitating membrane bleb stiffness using AFM force spectroscopy and an optical sideview setup.
AFM-based force spectroscopy in combination with optical microscopy is a powerful tool for investigating cell mechanics and adhesion on the single cell level. However, standard setups featuring an AFM mounted on an inverted light microscope only provide a bottom view of cell and AFM cantilever but cannot visualize vertical cell shape changes, for instance occurring during motile membrane blebbing. Here, we have integrated a mirror-based sideview system to monitor cell shape changes resulting from motile bleb behavior of Xenopus cranial neural crest (CNC) cells during AFM elasticity and adhesion measurements. Using the sideview setup, we quantitatively investigate mechanical changes associated with bleb formation and compared cell elasticity values recorded during membrane bleb and non-bleb events. Bleb protrusions displayed significantly lower stiffness compared to the non-blebbing membrane in the same cell. Bleb stiffness values were comparable to values obtained from blebbistatin-treated cells, consistent with the absence of a functional actomyosin network in bleb protrusions. Furthermore, we show that membrane blebs forming within the cell-cell contact zone have a detrimental effect on cell-cell adhesion forces, suggesting that mechanical changes associated with bleb protrusions promote cell-cell detachment or prevent adhesion reinforcement. Incorporating a sideview setup into an AFM platform therefore provides a new tool to correlate changes in cell morphology with results from force spectroscopy experiments.


Why differentiating between health system support and health system strengthening is needed.
There is increasing recognition that efforts to improve global health cannot be achieved without stronger health systems. Interpretation of health system strengthening (HSS) has varied widely however, with much of the focus to-date on alleviating input constraints, whereas less attention has been given to other performance drivers. It is important to distinguish activities that support the health system, from ones that strengthen the health system. Supporting the health system can include any activity that improves services, from distributing mosquito nets to procuring medicines. These activities improve outcomes primarily by increasing inputs. Strengthening the health system is accomplished by more comprehensive changes to performance drivers such as policies and regulations, organizational structures, and relationships across the health system to motivate changes in behavior and/or allow more effective use of resources to improve multiple health services. Even organizations that have made significant investments in health systems have not provided guidance on what HSS entails. While both supporting and strengthening are important and necessary, it is nonetheless important to make a distinction. If activities fail to produce improvements in system performance because they were incorrectly labeled as system strengthening, the value of HSS investments could quickly be discredited. Not distinguishing supportive activities from strengthening ones will lead to unmet expectations of stronger health systems, as well as neglect of critical system strengthening activities. Distinguishing between these two types of activities will improve programming impact.


Vagal sparing surgical technique but not stoma size affects body weight loss in rodent model of gastric bypass.
The aim of this study was to evaluate whether gastric bypass with or without vagal preservation resulted in a different outcome. Body weight, food intake and postprandial peptide YY (PYY) and glucagon-like peptide (GLP-1) levels were compared between gastric bypass (n = 55) and sham-operated rats (n = 27) in three groups. In group 1 (n = 17), the vagal nerve was not preserved, while in group 2 the vagal nerve was preserved during gastric bypass (n = 10). In group 3, gastric bypass rats (n = 28) were randomised for either one of the two techniques. Rats in which the vagal nerve was preserved during gastric bypass showed a lower body weight (p < 0.001) and reduced food intake (p < 0.001) compared to rats in which the vagal nerve was not preserved during the gastric bypass operation. Levels of PYY and GLP-1 were significantly increased after gastric bypass compared to sham-operated controls (p < 0.05), but there was no difference between gastric bypass rats with and without vagal preservation. Differences in food intake and body weight were not related to the size of the gastro-jejunostomy in gastric bypass rats. There were no signs of malabsorption or inflammation after gastric bypass. We propose that the vagal nerve should be preserved during the gastric bypass operation as this might play an important role for the mechanisms that induce weight loss and reduce food intake in rats. In contrast, the gastro-jejunal stoma size was found to be of minor relevance.


Weight status and nonadherence to asthma maintenance therapy among children enrolled in a public drug insurance plan.
Objective: The pediatric obese-asthma phenotype is associated with poor control, perhaps because of medication nonadherence. This study aimed to assess whether weight status is associated with nonadherence in children prescribed new asthma maintenance therapies.Methods: A historical cohort was constructed from a clinical database linking individual patient and prescription data to Quebec's prescription claims registry. Children aged 2-18 years with specialist-diagnosed asthma who were newly prescribed one of the following maintenance controllers: leukotriene receptor antagonists (LTRA); low-dose inhaled corticosteroids (ICS); medium/high-dose ICS; or combination therapy (ICS with long-acting beta-2 agonists and/or LTRA), at the Asthma Center of the Montreal Children's Hospital from 2000-2007 were included. Primary nonadherence was defined as not claiming any prescriptions, whereas secondary nonadherence was measured with the proportion of prescribed days covered (PPDC ≤ 50%) among primary adherers over a 6-month follow-up period. A modified Poisson regression model served to estimate the effect of excess weight (BMI > 85th percentile) on primary and secondary nonadherence.Results: Approximately one third of patients were primary nonadherers and 60% took less than 50% of prescribed therapy. Excess weight was associated with a trend toward increased risk of primary nonadherence in children newly prescribed low-dose ICS (RR 1.53, 95%CI 0.94-2.49), and of secondary nonadherence in children initiating medium/high-dose ICS (RR 1.24; 95%CI 0.98-1.59).Conclusions: Excess weight status is a possible determinant of primary nonadherence in children initiating low-dose ICS and secondary nonadherence to higher-dose ICS regimens. This hypothesis-generating study suggests that nonadherence may be a potential contributor to higher morbidity in children with obese-asthma.


Postoperative atrial fibrillation prediction following isolated surgical aortic valve replacement.
Postoperative atrial fibrillation (POAF) is the most common complication following cardiac surgery, with increased risk of stroke and high mortality. Our aim was to identify patients at risk and to design a model that could predict POAF. In this single center study, we evaluated 1191 patients requiring isolated surgical aortic valve replacement between January 2000 and June 2014. The patients were followed during the early postoperative period until discharge. AF occurred in 342 patients (28.71%). Six variables associated with high arrhythmic risk [advanced age, body mass index, tricuspid regurgitation, prolonged ventilation, longer intensive care unit stay, and dilated left atrium (LA; volume ≥35 ml/m2)] were selected to create a multivariate prediction model. This model predicted POAF in 64.7% of cases, with a moderate discriminative power (AUC=0.65; p=0.001; 95% CI, 0.571-0.771). We also developed the CHAID (Chi-square automatic interaction detection) model showing multilevel interactions among risk factors for POAF. Age had the greatest discriminative power, with patients aged >68 years at a higher risk for POAF. In low-risk patients, the subgroup with dilated LA (volume ≥40 ml) was more prone to develop POAF. For the intermediate-risk group, history of AF was the next deciding variable, whereas for the high-risk group, it was tricuspid regurgitation (at least moderate). The multivariate logistic model has an acceptable predictive value. CHAID-derived model is a new tool that could be easily applied to identify patients requiring prophylactic regimens.


Evaluation of the efficacy and safety of changes in antiretroviral regimens for HIV-infected patients.
Antiretroviral therapy is now available for HIV-infected patients, and so-called highly active antiretroviral therapy (HAART) now makes it possible to strongly suppress viral proliferation and restore immunity. However, the development of new drugs and regimens for HAART is still in progress, with the aim of overcoming a number of associated problems. For this purpose, changes in the prescribed anti-HIV drugs are often made. In the present study, we attempted to clarify the actual effects of such treatment modifications in patients who had been started on HAART. We retrospectively investigated HIV-infected patients who had been started on HAART at Kitasato University Hospital between April 1997 and March 2013. The patients' backgrounds, characteristics and laboratory data were established from the hospital medical records. The total follow-up time was 447.3 person-years. The patients remained on their initial regimen for a median period of 2040 days, and 39 patients took a second regimen for a median of 2714 days. There was no treatment failure due to regimen change. The reason for the regimen change was adverse effects in 49 cases, poor adherence/virological failure in 4, immunological failure in 3, patient request in 2, and proposals made by health care workers, or for simplification, in 11. The number of patients who required regimen change due to renal dysfunction showed a gradual increase. The number of times anti-HIV drugs were taken per day was not altered when the regimen changed, being mainly once or twice a day. In the present study, there were no instances of treatment failure due to regimen change. Through appropriate regimen change, it is possible to avoid serious adverse effects, and to improve patient adherence. Further adverse effects associated with long-term antiretroviral therapy, and reduction of adherence through medication fatigue should be considered. Drug selection and regimen change should be considered in relation to long-term prognosis.


Small RNAs in early mammalian development: from gametes to gastrulation.
Small non-coding RNAs, including microRNAs (miRNAs), endogenous small interfering RNAs (endo-siRNAs) and Piwi-interacting RNAs (piRNAs), play essential roles in mammalian development. The function and timing of expression of these three classes of small RNAs differ greatly. piRNAs are expressed and play a crucial role during male gametogenesis, whereas endo-siRNAs are essential for oocyte meiosis. By contrast, miRNAs are ubiquitously expressed in somatic tissues and function throughout post-implantation development. Surprisingly, however, miRNAs are non-essential during pre-implantation embryonic development and their function is suppressed during oocyte meiosis. Here, we review the roles of small non-coding RNAs during the early stages of mammalian development, from gamete maturation through to gastrulation.


Exponential model for analysis of heart rate responses and autonomic cardiac modulation during different intensities of physical exercise.
The aim of this study was to compare the heart rate (HR) dynamics and variability before and after high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) protocols with workloads based on treadmill workload at which maximal oxygen uptake was achieved ( WL V ˙ O 2 max ) . Ten participants performed cardiopulmonary exercise testing (CPET) to obtain oxygen uptake ( WL V ˙ O 2 max ) . All training protocols were performed on a treadmill, with 0% grade, and had similar total distance. The MICT was composed by 21 min at 70% of WL V ˙ O 2 max . The first HIIT protocol (HIIT-30 : 30) was composed by 29 repetitions of 30 s at 100% of s V ˙ O 2 max and the second HIIT protocol (HIIT-4 : 3) was composed by three repetitions of 4 min at 90% of WL V ˙ O 2 max . Before, during and after each training protocol, HR dynamics and variability (HRV) were analysed by standard kinetics and linear (time and frequency domains). The repeated measures analysis of variance indicated that the HR dynamics, which characterizes the speed of HR during the rest to exercise transition, was statistically (p < 0.05) slower during MICT in comparison to both HIIT protocols. The HRV analysis, which characterizes the cardiac autonomic modulation during the exercise recovery, was statistically higher in HIIT-4 : 3 in comparison to MICT and HIIT-30 : 30 protocols (p < 0.005 and p = 0.012, respectively), suggesting that the HIIT-4 : 3 induced higher sympathetic and lower parasympathetic modulation during exercise in comparison to the other training protocols. In conclusion, HIIT-4 : 3 demonstrated post-exercise sympathetic hyperactivity and a higher HRpeak, while the HIIT-30 : 30 and MICT resulted in better HRV and HR in the exercise-recovery transition. The cardiac autonomic balance increased in HIIT-30 : 30 while HIIT-4 : 3 induced sympathetic hyperactivity and cardiac overload.


Corticosteroid-Related Adverse Events Systematically Increase with Corticosteroid Dose in Noninfectious Intermediate, Posterior, or Panuveitis: Post Hoc Analyses from the VISUAL-1 and VISUAL-2 Trials.
Chronic use of corticosteroids for the treatment of uveitis has been linked with drug-associated toxicity and adverse events (AEs). This study examines the association between corticosteroid dosage and incidence rates of corticosteroid-related AEs. A post hoc analysis of the VISUAL-1 and VISUAL-2 placebo-controlled clinical trials. The clinical trials consisted of adults with active (VISUAL-1) and inactive (VISUAL-2) noninfectious intermediate, posterior, and panuveitis. Patients were randomized to receive adalimumab or placebo and underwent a protocol-defined mandatory taper to discontinue their oral corticosteroids. Adverse event data were collected at each visit and included an assessment of the corticosteroid relationship by the investigator. A longitudinal Poisson regression model was estimated controlling for time-dependent corticosteroid dose, age, sex, prior oral corticosteroid dose, prior topical corticosteroid use, and concomitant immunosuppressive drug use. Only patients randomized to placebo were considered. The primary outcome measure was the frequency of AEs. The incidence rates of corticosteroid-related AEs among placebo patients during the prednisone treatment period in VISUAL-1 was statistically higher than after discontinuation (454.2 per 100 patient-years [PY] vs. 36.1 per 100 PY, incident rate ratio = 12.6, P < 0.001). Incidence rate ratios among VISUAL-2 patients were similarly high (317.5 per 100 PY vs. 41.1 per 100 PY, incident rate ratio = 7.7, P < 0.001). Based on the Poisson multivariate longitudinal Generalized Estimating Equation (GEE) model, each 10 mg increase in prednisone dose is associated with a 1.5- and 2.6-fold increase (P < 0.001 and P < 0.001) in the rate of corticosteroid-related AEs in VISUAL-1 and VISUAL-2, respectively. This implies in turn that a patient with active uveitis taking 60 mg/day of prednisone will experience, on average, an additional 10.1 (95% confidence interval (CI), 6.3-14.5; P < 0.001) corticosteroid-related AEs per year compared with a patient taking 10 mg/day, whereas a patient with inactive uveitis taking 35 mg/day of prednisone will experience, on average, an additional 23.5 (95% CI, 7.6-52.7; P = 0.05) corticosteroid-related AEs per year compared with a patient taking 10 mg/day. Evidence from VISUAL-1 and VISUAL-2 suggests that the incidence rates of corticosteroid-related AEs increase systematically with corticosteroid dose.