INHALEd nebulised unfractionated HEParin for the treatment of hospitalised patients with COVID-19 (INHALE-HEP): Protocol and statistical analysis plan for an investigator-initiated international metatrial of randomised studies.
Inhaled nebulised unfractionated heparin (UFH) has a strong scientific and biological rationale that warrants urgent investigation of its therapeutic potential in patients with COVID-19. UFH has antiviral effects and prevents the SARS-CoV-2 virus' entry into mammalian cells. In addition, UFH has significant anti-inflammatory and anticoagulant properties, which limit progression of lung injury and vascular pulmonary thrombosis. The INHALEd nebulised unfractionated HEParin for the treatment of hospitalised patients with COVID-19 (INHALE-HEP) metatrial is a prospective individual patient data analysis of on-going randomised controlled trials and early phase studies. Individual studies are being conducted in multiple countries. Participating studies randomise adult patients admitted to the hospital with confirmed SARS-CoV-2 infection, who do not require immediate mechanical ventilation, to inhaled nebulised UFH or standard care. All studies collect a minimum core dataset. The primary outcome for the metatrial is intubation (or death, for patients who died before intubation) at day 28. The secondary outcomes are oxygenation, clinical worsening and mortality, assessed in time-to-event analyses. Individual studies may have additional outcomes. We use a Bayesian approach to monitoring, followed by analysing individual patient data, outcomes and adverse events. All analyses will follow the intention-to-treat principle, considering all participants in the treatment group to which they were assigned, except for cases lost to follow-up or withdrawn. The metatrial is registered at ClinicalTrials.gov ID NCT04635241. Each contributing study is individually registered and has received approval of the relevant ethics committee or institutional review board. Results of this study will be shared with the World Health Organisation, published in scientific journals and presented at scientific meetings.


Behaviorally bisexual men and their risk behaviors with men and women.
Gay and bisexual men are often treated as a homogenous group; however, there may be important differences between them. In addition, behaviorally bisexual men are a potential source of HIV infection for heterosexual women. In this study, we compared 97 men who have sex with men only (MSM) to 175 men who have sex with men and women (MSMW). We also compared the 175 MSMW to 772 men who have sex with women only (MSW). Bivariate and multiple logistic regression analyses were performed to assess correlates of MSMW risk behaviors with men and with women as well as whether MSMW, compared with MSW, engaged in more risky behaviors with women. Compared with MSM, MSMW were less likely to be HIV-positive or to engage in unprotected receptive anal intercourse. In contrast, MSMW were more likely than MSW to be HIV-positive and to engage in anal intercourse with their female partners; however, rates of unprotected anal intercourse were similar. The study findings suggest that there may be important differences in HIV risk behaviors and HIV prevalence between MSM and MSMW as well as between MSMW and MSW.


Toxoplasma gondii: from animals to humans.
Toxoplasmosis is one of the more common parasitic zoonoses world-wide. Its causative agent, Toxoplasma gondii, is a facultatively heteroxenous, polyxenous protozoon that has developed several potential routes of transmission within and between different host species. If first contracted during pregnancy, T. gondii may be transmitted vertically by tachyzoites that are passed to the foetus via the placenta. Horizontal transmission of T. gondii may involve three life-cycle stages, i.e. ingesting infectious oocysts from the environment or ingesting tissue cysts or tachyzoites which are contained in meat or primary offal (viscera) of many different animals. Transmission may also occur via tachyzoites contained in blood products, tissue transplants, or unpasteurised milk. However, it is not known which of these routes is more important epidemiologically. In the past, the consumption of raw or undercooked meat, in particular of pigs and sheep, has been regarded as a major route of transmission to humans. However, recent studies showed that the prevalence of T. gondii in meat-producing animals decreased considerably over the past 20 years in areas with intensive farm management. For example, in several countries of the European Union prevalences of T. gondii in fattening pigs are now <1%. Considering these data it is unlikely that pork is still a major source of infection for humans in these countries. However, it is likely that the major routes of transmission are different in human populations with differences in culture and eating habits. In the Americas, recent outbreaks of acute toxoplasmosis in humans have been associated with oocyst contamination of the environment. Therefore, future epidemiological studies on T. gondii infections should consider the role of oocysts as potential sources of infection for humans, and methods to monitor these are currently being developed. This review presents recent epidemiological data on T. gondii, hypotheses on the major routes of transmission to humans in different populations, and preventive measures that may reduce the risk of contracting a primary infection during pregnancy.


Assessing outcomes of the educational program of the West Virginia School of Osteopathic Medicine.
In education, "assessment" refers to the ongoing, systematic process of identification of goals, development of methods for measuring progress toward attaining those goals, and use of resulting information to make revisions in educational programs. Recently, state-review programs and accrediting agencies have focused review efforts on outcomes of the educational process. The West Virginia School of Osteopathic Medicine has developed a successful outcomes assessment program that includes information regarding graduates' activities. The school is now considering new programs regarding more comprehensive assessment of clinical outcomes.


A comparison of the effects of quetiapine ('seroquel') and haloperidol in schizophrenic patients with a history of and a demonstrated, partial response to conventional antipsychotic treatment. PRIZE Study Group.
Quetiapine ('Seroquel') is a well-tolerated, novel, atypical antipsychotic with consistent efficacy in the treatment of schizophrenia. To date, no clinical studies have evaluated the effect of quetiapine in patients who only partially respond to conventional antipsychotics, yet this type of patient is most frequently seen by psychiatrists. Therefore, this international, multicentre, double-blind study was conducted to compare the efficacy and tolerability of 8 weeks' treatment of quetiapine 600 mg/day with haloperidol 20 mg/day in 288 patients who had a history of partial response to conventional antipsychotics and displayed a partial or no response to 1 month of fluphenazine (20 mg/day) treatment. Patients on quetiapine tended to have greater improvement than those on haloperidol in the primary efficacy measure, mean Positive and Negative Symptom Scale (PANSS) score, after 4 weeks' treatment (-9.05, -5.82, respectively, P = 0.061) and at study end (-11.50, -8.87, respectively, P = 0.234). Similarly, there was a trend towards patients on quetiapine demonstrating greater improvements in the secondary efficacy measures (Clinical Global Impression, PANSS subscale and Brief Psychiatric Rating Scale scores) [week 4 (baseline) to week 12 (end)], but the difference between treatments did not reach significance. Significantly more patients on quetiapine than on haloperidol showed a clinical response-patient response rates, defined as > 20% reduction in PANSS total score between weeks 4 and 12, were 52.2% for quetiapine and 38.0% for haloperidol (P = 0.043). Patients receiving quetiapine required less anticholinergic medication (P < 0.011), had greater reduction in extrapyramidal symptoms (EPS) (P = 0.005) and fewer treatment-emergent EPS-related adverse events compared to those on haloperidol (P < 0.001). Serum prolactin concentrations were elevated at the end of fluphenazine treatment in 73% of patients. Between weeks 4 and 12, elevated serum prolactin concentrations significantly decreased in quetiapine-treated patients compared to those receiving haloperidol (P < 0.001). At the end of quetiapine treatment, 83% of patients had normal prolactin levels while only 21% of patients receiving haloperidol were within the normal range. These results suggest that quetiapine may make a valuable contribution to the management of patients with a history of partial response to conventional antipsychotics.


Which Game Narratives Do Adolescents of Different Gameplay and Sociodemographic Backgrounds Prefer? A Mixed-Methods Analysis.
Objective: The aim of this study was to investigate which narrative elements of digital game narratives are preferred by the general adolescent population, and to examine associations with gender, socioeconomic status (SES), and gameplay frequency. Further, the study aims to discuss how results can be translated to serious digital games. Materials and Methods: Adolescents were recruited through school to complete a survey on narrative preferences in digital games. The survey included questions on sociodemographic information, frequency of gameplay, and an open-ended question on what could be an appealing narrative for them. Data were analyzed in a mixed-methods approach, using thematic analysis and chi-square analyses to determine narrative preferences and the associations between game narrative elements and player characteristics (gender, SES, and frequency of gameplay). Results: The sample consisted of 446 adolescents (12-15 years old) who described 30 narrative subthemes. Preferences included human characters as protagonists; nonhuman characters only as antagonists; realistic settings, such as public places or cities; and a strong conflict surrounding crime, catastrophe, or war. Girls more often than boys defined characters by their age, included avatars, located the narrative in private places, developed profession-related skills, and included a positive atmosphere. Adolescents of nonacademic education more often than adolescents of academic education defined characters by criminal actions. Infrequent players more often included human characters defined by their age than frequent players. After performing a Bonferroni correction, narrative preferences for several gender differences remained. Conclusion: Different narrative elements related to subgroups of adolescents by gender, SES, and frequency of gameplay. Customization of narratives in serious digital health games should be warranted for boys and girls; yet, further research is needed to specify how to address girls in particular.


Slow-release carbohydrates: growing evidence on metabolic responses and public health interest. Summary of the symposium held at the 12th European Nutrition Conference (FENS 2015).
To draw attention to the necessity of considering differences in the digestibility of carbohydrates, and more specifically of starch, a symposium was held at the 12th European Nutrition Conference (FENS), which took place in Berlin from October 20 to 23, 2015. The purpose of this session was to present the consolidated knowledge and recent advances regarding the relationship between slow-release carbohydrates, metabolic responses, and public health issues. Three main topics were presented: 1) the definition of, sources of, and recognised interest in the glycaemic response to slowly digestible starch (SDS); 2) clinical evidence regarding the physiological effects of slow-release carbohydrates from cereal foods; and 3) interest in reducing the postprandial glycaemic response to help prevent metabolic diseases. Foods with the highest SDS content induce the lowest glycaemic responses, as the starch is protected from gelatinisation during processing. In humans, high-SDS food consumption induces slower glucose release, lower postprandial insulinaemia, and stimulation of gut hormones. Moreover, postprandial hyperglycaemia is an independent risk factor for type two diabetes mellitus (T2DM) and cardiovascular disease (CVD). Therefore, given the plausible aetiologic mechanisms, we argue that postprandial glucose levels are relevant for health and disease and represent a meaningful target for intervention, for example, through dietary factors. This symposium was organised by Mondelez International R&D.


Coding practice in national and regional kidney biopsy registries.
Kidney biopsy registries all over the world benefit research, teaching and health policy. Comparison, aggregation and exchange of data is however greatly dependent on how registration and coding of kidney biopsy diagnoses are performed. This paper gives an overview over kidney biopsy registries, explores how these registries code kidney disease and identifies needs for improvement of coding practice. A literature search was undertaken to identify biopsy registries for medical kidney diseases. These data were supplemented with information from personal contacts and from registry websites. A questionnaire was sent to all identified registries, investigating age of registries, scope, method of coding, possible mapping to international terminologies as well as self-reported problems and suggestions for improvement. Sixteen regional or national kidney biopsy registries were identified, of which 11 were older than 10 years. Most registries were located either in Europe (10/16) or in Asia (4/16). Registries most often use a proprietary coding system (12/16). Only a few of these coding systems were mapped to SNOMED CT (1), older SNOMED versions (2) or ERA-EDTA PRD (3). Lack of maintenance and updates of the coding system was the most commonly reported problem. There were large gaps in the global coverage of kidney biopsy registries. Limited use of international coding systems among existing registries hampers interoperability and exchange of data. The study underlines that the use of a common and uniform coding system is necessary to fully realize the potential of kidney biopsy registries.


Helicobacter pylori antimicrobial resistance and antibiotic consumption in the low-resource Central America setting.
Antimicrobial resistance is a global public health problem, particularly in low- and middle-income countries (LMICs), where antibiotics are often obtained without a prescription. H. pylori antimicrobial resistance patterns are informative for patient care and gastric cancer prevention programs, have been shown to correlate with general antimicrobial consumption, and may guide antimicrobial stewardship programs in LMICs. We report H. pylori resistance and antimicrobial utilization patterns for western Honduras, representative of rural Central America. In the context of the western Honduras gastric cancer epidemiology initiative, gastric biopsies from 189 patients were studied for culture and resistance patterns. Antimicrobial utilization was investigated for common H. pylori treatment regimens from regional public (7 antimicrobials) and national private (4 antimicrobials) data, analyzed in accordance with WHO anatomical therapeutic chemical defined daily doses (DDD) method and expressed as DDD/1000 inhabitants per day (DID) and per year (DIY). H. pylori was successfully cultured from 116 patients (56% males, mean age: 54), and nearly all strains were cagA+ and vacAs1m1+ positive (99% and 90.4%, respectively). Unexpectedly, high resistance was noted for levofloxacin (20.9%) and amoxicillin (10.7%), while metronidazole (67.9%) and clarithromycin (11.2%) were similar to data from Latin America. Significant associations with age, gender, or histology were not noted, with the exception of levofloxacin (28%, P = 0.01) in those with histology limited to non-atrophic gastritis. Total antimicrobial usage in western Honduras of amoxicillin (17.3 DID) and the quinolones had the highest relative utilizations compared with other representative nations. We observed significant H. pylori resistance to amoxicillin and levofloxacin in the context of high community antimicrobial utilization. This has implications in Central America for H. pylori treatment guidelines as well as antimicrobial stewardship programs.


Common west African HLA antigens are associated with protection from severe malaria.
A large case-control study of malaria in West African children shows that a human leucocyte class I antigen (HLA-Bw53) and an HLA class II haplotype (DRB1*1302-DQB1*0501), common in West Africans but rare in other racial groups, are independently associated with protection from severe malaria. In this population they account for as great a reduction in disease incidence as the sickle-cell haemoglobin variant. These data support the hypothesis that the extraordinary polymorphism of major histocompatibility complex genes has evolved primarily through natural selection by infectious pathogens.


Assessment of factors affecting MRI measurement of intracranial volume changes and elastance index.
Non-invasive measurement of cerebral hydrodynamic parameters may be of great use in the study and treatment of conditions involving abnormal intracranial pressure (ICP). ICP can be assessed indirectly in humans by calculation of intracranial volume changes and elastance using MRI. These two hydrodynamic parameters are derived from mathematical analysis of the relationships between blood flow to and from the brain, CSF flow and heart rate. This paper examines the variability in these three critical subject specific parameters. Three healthy adult males had measurements of blood and CSF flow to and from the brain made on three occasions. Inter-individual variation in vasculature and intra-individual variation in heart rates led to modest repeatability (defined as standard deviation divided by the mean) of intracranial volume change and elastance index in two subjects and poor repeatability in the third. Particular care is required in the selection of image planes and the blood vessels to include, and the analysis is very sensitive to differences in heart-rate. Because of these difficulties, caution is required in the interpretation of results obtained from a single examination. Techniques and methods that may reduce the variability in the derived hydrodynamic parameters are discussed.


Shotgun proteomics reveals physiological response to ocean acidification in Crassostrea gigas.
Ocean acidification as a result of increased anthropogenic CO2 emissions is occurring in marine and estuarine environments worldwide. The coastal ocean experiences additional daily and seasonal fluctuations in pH that can be lower than projected end-of-century open ocean pH reductions. In order to assess the impact of ocean acidification on marine invertebrates, Pacific oysters (Crassostrea gigas) were exposed to one of four different p CO2 levels for four weeks: 400 μatm (pH 8.0), 800 μatm (pH 7.7), 1000 μatm (pH 7.6), or 2800 μatm (pH 7.3). At the end of the four week exposure period, oysters in all four p CO2 environments deposited new shell, but growth rate was not different among the treatments. However, micromechanical properties of the new shell were compromised by elevated p CO2. Elevated p CO2 affected neither whole body fatty acid composition, nor glycogen content, nor mortality rate associated with acute heat shock. Shotgun proteomics revealed that several physiological pathways were significantly affected by ocean acidification, including antioxidant response, carbohydrate metabolism, and transcription and translation. Additionally, the proteomic response to a second stress differed with p CO2, with numerous processes significantly affected by mechanical stimulation at high versus low p CO2 (all proteomics data are available in the ProteomeXchange under the identifier PXD000835). Oyster physiology is significantly altered by exposure to elevated p CO2, indicating changes in energy resource use. This is especially apparent in the assessment of the effects of p CO2 on the proteomic response to a second stress. The altered stress response illustrates that ocean acidification may impact how oysters respond to other changes in their environment. These data contribute to an integrative view of the effects of ocean acidification on oysters as well as physiological trade-offs during environmental stress.


Colonization, competition, and dispersal of pathogens in fluid flow networks.
The colonization of bacteria in complex fluid flow networks, such as those found in host vasculature, remains poorly understood. Recently, it was reported that many bacteria, including Bacillus subtilis [1], Escherichia coli [2], and Pseudomonas aeruginosa [3, 4], can move in the opposite direction of fluid flow. Upstream movement results from the interplay between fluid shear stress and bacterial motility structures, and such rheotactic-like behavior is predicted to occur for a wide range of conditions [1]. Given the potential ubiquity of upstream movement, its impact on population-level behaviors within hosts could be significant. Here, we find that P. aeruginosa communities use a diverse set of motility strategies, including a novel surface-motility mechanism characterized by counter-advection and transverse diffusion, to rapidly disperse throughout vasculature-like flow networks. These motility modalities give P. aeruginosa a selective growth advantage, enabling it to self-segregate from other human pathogens such as Proteus mirabilis and Staphylococcus aureus that outcompete P. aeruginosa in well-mixed non-flow environments. We develop a quantitative model of bacterial colonization in flow networks, confirm our model in vivo in plant vasculature, and validate a key prediction that colonization and dispersal can be inhibited by modifying surface chemistry. Our results show that the interaction between flow mechanics and motility structures shapes the formation of mixed-species communities and suggest a general mechanism by which bacteria could colonize hosts. Furthermore, our results suggest novel strategies for tuning the composition of multi-species bacterial communities in hosts, preventing inappropriate colonization in medical devices, and combatting bacterial infections.


Activity of oxantel pamoate monotherapy and combination chemotherapy against Trichuris muris and hookworms: revival of an old drug.
It is widely recognized that only a handful of drugs are available against soil-transmitted helminthiasis, all of which are characterized by a low efficacy against Trichuris trichiura, when administered as single doses. The re-evaluation of old, forgotten drugs is a promising strategy to identify alternative anthelminthic drug candidates or drug combinations. We studied the activity of the veterinary drug oxantel pamoate against Trichuris muris, Ancylostoma ceylanicum and Necator americanus in vitro and in vivo. In addition, the dose-effect of oxantel pamoate combined with albendazole, mebendazole, levamisole, pyrantel pamoate and ivermectin was studied against T. muris in vitro and additive or synergistic combinations were followed up in vivo. We calculated an ED50 of 4.7 mg/kg for oxantel pamoate against T. muris in mice. Combinations of oxantel pamoate with pyrantel pamoate behaved antagonistically in vitro (combination index (CI) = 2.53). Oxantel pamoate combined with levamisole, albendazole or ivermectin using ratios based on their ED50s revealed antagonistic effects in vivo (CI = 1.27, 1.90 and 1.27, respectively). A highly synergistic effect (CI = 0.15) was observed when oxantel pamoate-mebendazole was administered to T. muris-infected mice. Oxantel pamoate (10 mg/kg) lacked activity against Ancylostoma ceylanicum and Necator americanus in vivo. Our study confirms the excellent trichuricidal properties of oxantel pamoate. Since the drug lacks activity against hookworms it is necessary to combine oxantel pamoate with a partner drug with anti-hookworm properties. Synergistic effects were observed for oxantel pamoate-mebendazole, hence this combination should be studied in more detail. Since, of the standard drugs, albendazole has the highest efficacy against hookworms, additional investigations on the combination effect of oxantel pamoate-albendazole should be launched.


Riboflavin kinase couples TNF receptor 1 to NADPH oxidase.
Reactive oxygen species (ROS) produced by NADPH oxidase function as defence and signalling molecules related to innate immunity and various cellular responses. The activation of NADPH oxidase in response to plasma membrane receptor activation depends on the phosphorylation of cytoplasmic oxidase subunits, their translocation to membranes and the assembly of all NADPH oxidase components. Tumour necrosis factor (TNF) is a prominent stimulus of ROS production, but the molecular mechanisms by which TNF activates NADPH oxidase are poorly understood. Here we identify riboflavin kinase (RFK, formerly known as flavokinase) as a previously unrecognized TNF-receptor-1 (TNFR1)-binding protein that physically and functionally couples TNFR1 to NADPH oxidase. In mouse and human cells, RFK binds to both the TNFR1-death domain and to p22(phox), the common subunit of NADPH oxidase isoforms. RFK-mediated bridging of TNFR1 and p22(phox) is a prerequisite for TNF-induced but not for Toll-like-receptor-induced ROS production. Exogenous flavin mononucleotide or FAD was able to substitute fully for TNF stimulation of NADPH oxidase in RFK-deficient cells. RFK is rate-limiting in the synthesis of FAD, an essential prosthetic group of NADPH oxidase. The results suggest that TNF, through the activation of RFK, enhances the incorporation of FAD in NADPH oxidase enzymes, a critical step for the assembly and activation of NADPH oxidase.


The underrecognized burden of influenza in young children.
The disease burden of influenza infection among children is not well established. We conducted a population-based surveillance of medical visits associated with laboratory-confirmed influenza. Eligible children were younger than five years of age, resided in three U.S. counties, and had a medical visit for an acute respiratory tract infection or fever. Nasal and throat swabs were tested for the influenza virus by viral culture and polymerase-chain-reaction assay. Epidemiologic data were collected from parental surveys and chart reviews. Children who were hospitalized were enrolled prospectively from 2000 through 2004. Population-based rates of hospitalizations associated with influenza were calculated. Children who were seen in selected pediatric clinics and emergency departments during two influenza seasons (2002-2003 and 2003-2004) were systematically enrolled. The rates of visits to clinics and emergency departments associated with influenza were estimated. The average annual rate of hospitalization associated with influenza was 0.9 per 1000 children. The estimated burden of outpatient visits associated with influenza was 50 clinic visits and 6 emergency department visits per 1000 children during the 2002-2003 season and 95 clinic visits and 27 emergency department visits per 1000 children during the 2003-2004 season. Few children who had laboratory-confirmed influenza were given a diagnosis of influenza by the treating physician in the inpatient (28 percent) or outpatient (17 percent) settings. Among young children, outpatient visits associated with influenza were 10 to 250 times as common as hospitalizations. Few influenza infections were recognized clinically.


Size and form in efficient transportation networks.
Many biological processes, from cellular metabolism to population dynamics, are characterized by allometric scaling (power-law) relationships between size and rate. An outstanding question is whether typical allometric scaling relationships--the power-law dependence of a biological rate on body mass--can be understood by considering the general features of branching networks serving a particular volume. Distributed networks in nature stem from the need for effective connectivity, and occur both in biological systems such as cardiovascular and respiratory networks and plant vascular and root systems, and in inanimate systems such as the drainage network of river basins. Here we derive a general relationship between size and flow rates in arbitrary networks with local connectivity. Our theory accounts in a general way for the quarter-power allometric scaling of living organisms, recently derived under specific assumptions for particular network geometries. It also predicts scaling relations applicable to all efficient transportation networks, which we verify from observational data on the river drainage basins. Allometric scaling is therefore shown to originate from the general features of networks irrespective of dynamical or geometric assumptions.


Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity.
The possibility of DNA degradation is of concern to all involved in the storage of DNA, whether for diagnostic or research purposes. Many DNA banks are at present maintained at low temperatures, but optimum conditions for storage and handling have yet to be fully assessed. Both DNA and fresh blood have been subjected to repeated cycles of freezing and thawing and DNA extracted from the blood. DNA yield has been established and integrity examined by digestion, electrophoresis, and Southern blot analysis using DNA fingerprinting techniques. No degradation of DNA could be detected using these techniques; however, DNA yield was shown to be adversely affected by freezing, with yield reduced by more than 25% in blood samples frozen only once.


Treatment of mantle-cell lymphoma with Rituximab (chimeric monoclonal anti-CD20 antibody): analysis of factors associated with response.
A retrospective analysis was performed to delineate the factors associated with response, and to determine the duration of response, in 87 patients with CD20-positive mantle-cell lymphoma (MCL) treated with Rituximab (chimeric monoclonal anti-CD20 antibody) in two prior studies. Patients with newly-diagnosed MCL (MCL1, n = 37), and previously-treated MCL (MCL2, n = 50), received single-agent Rituximab, in the context of two multicentre clinical studies using different schedules and doses, conducted in 1996 and 1997. A follow-up analysis was performed at the end of 1998, including all 81 patients who completed therapy. Statistical modeling of factors associated with response was performed using ordered logistic regression. The duration of complete (CR) and partial response (PR), and the time to disease progression (TTP), were also derived. The overall response rate (RR) was 34% (30 of 87) (81 evaluable patients, RR 37%; CR 14%), and was equivalent for MCL1 and MCL2. On univariate analysis, elevated LDH (P = 0.004); prior therapy with alkylating agents (P = 0.01) or fludarabine phosphate (P = 0.04); WHO performance status = 2 (P = 0.02); MCL2 refractory to last prior therapy (P = 0.04); and splenomegaly (P = 0.04), each at the time of treatment with Rituximab, were significantly associated with a lower RR. On multivariate analysis, only LDH (P = 0.007) and prior alkylating agents (P = 0.03) retained statistical significance. At a median follow-up of 1.4 years, the median TTP was 7 months. The median duration of response was one year, and was significantly longer for patients achieving CR vs. PR (P = 0.04). Rituximab is active in MCL, and can induce complete responses in a minority of patients. Elevated LDH at the time of therapy, and prior therapy with alkylating agents, are associated with a significantly lower RR. The duration of response of one year is similar to that previously reported in follicular lymphoma.


Isolation and characterization of cell subpopulations from the monkey corpus luteum of the menstrual cycle.
This study was designed to test the hypothesis that the corpus luteum of primate species consists of cell subpopulations that differ in physical characteristics, function, and regulation by endocrine and paracrine factors. The corpus luteum (n = 25) was removed from rhesus monkeys at the mid-luteal phase of the menstrual cycle (Days 7-8 after the surge of luteinizing hormone, LH) and enzymatically dispersed. Freshly dispersed cells were analyzed and sorted on the basis of their forward and 90 degrees light scatter (FLS and 90LS, respectively) properties using an EPICS C flow cytometer. Freshly dispersed and sorted cells were fixed, stained histochemically for the presence of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), and measured to determine their diameters. Freshly dispersed (MIX) and sorted cells from corpora lutea during the early (Days 4-5 after the LH surge; n = 4) and mid-luteal phases of the cycle were incubated in vitro and steroid production was assessed. The size distribution of dispersed cells revealed four peaks that corresponded to small (10-15 microns in diameter) 3 beta-HSD-negative, and small, medium (16-20 microns), and large (greater than 20 microns) 3 beta-HSD-positive cells. Analysis of dispersed cells for FLS and 90LS demonstrated two continua (C alpha and C beta). C alpha contained single cells and cell clusters; 99.7 +/- 0.3% (n = 3) of the cells were less than or equal to 15 microns in diameter and 96.7 +/- 0.3% were 3 beta-HSD-negative. C alpha cells produced low levels of progesterone (0.2 +/- 0.1 ng/ml per 5 x 10(4) cells; n = 3) in vitro under basal conditions. C beta consisted of single cells from 10 microns to 40 microns in diameter and contained the lipid-filled and 3 beta-HSD-positive cells. Two regions (R1 and R3) of C beta were defined and their cells separated. In R1, 96 +/- 2% (n = 3) of the cells had diameters of less than or equal to 15 microns, whereas 82 +/- 4% (n = 3) of those in R3 were greater than or equal to 20 microns. Basal progesterone production by R3 cells from early luteal phase of the cycle was 12 times greater than that by R1 cells (n = 3 per group).(ABSTRACT TRUNCATED AT 400 WORDS)


Expressive writing as a brief intervention for reducing drinking intentions.
The present study examined the effectiveness of expressive writing in reducing drinking behavior. We expected that students prompted to write about negative drinking experiences would show greater decreases in future drinking intentions compared to the neutral and the positive writing conditions. We also expected that decreases in drinking intentions following the writing prompts might differ based on current drinking and AUDIT scores. Participants included 200 (76% female) undergraduates who completed measures of their current drinking behavior. They were then randomly assigned to either write about: a time when they had a lot to drink that was a good time (Positive); a time when they had a lot to drink that was a bad time (Negative); or their first day of college (Neutral), followed by measures assessing intended drinking over the next three months. Results revealed that participants intended to drink significantly fewer drinks per week and engage in marginally fewer heavy drinking occasions after writing about a negative drinking occasion when compared to control. Interactions provided mixed findings suggesting that writing about a positive event was associated with higher drinking intentions for heavier drinkers. Writing about a negative event was associated with higher intentions among heavier drinkers, but lower intentions among those with higher AUDIT scores. This research builds on previous expressive writing interventions by applying this technique to undergraduate drinkers. Preliminary results provide some support for this innovative strategy but also suggest the need for further refinement, especially with heavier drinkers.


Concentric circular trajectory sampling for super-resolution and image mosaicing.
Ubiquitous applications in diverse fields motivate large-area sampling, super-resolution (SR) and image mosaicing. However, conventional translational sampling has drawbacks including laterally constrained variations between samples. Meanwhile, existing rotational sampling methods do not consider the uniformity of sampling points in Cartesian coordinates, resulting in additional distortion errors in sampled images. We design a novel optimized concentric circular trajectory sampling (OCCTS) method to acquire image information uniformly at fast sampling speeds. The sampling method allows multiple low-resolution images for conventional SR algorithms to be acquired by adding small variations in the angular dimension. Experimental results demonstrate that OCCTS can beat comparable CCTS methods that lack optimized sampling densities by reducing sampling time by more than 11.5% while maintaining 50% distortion error reduction. The SR quality of OCCTS has at least 5.2% fewer distortion errors than the comparable CCTS methods. This paper is the first, to the best of our knowledge, to present an OCCTS method for SR and image mosaicing.


Pharmacokinetics and pharmacodynamics of valproate analogs in rats. III. Pharmacokinetics of valproic acid, cyclohexanecarboxylic acid, and 1-methyl-1-cyclohexanecarboxylic acid in the bile-exteriorized rat.
The pharmacokinetics of valproic acid (VPA) and its structural analogs cyclohexanecarboxylic acid (CCA) and 1-methyl-1-cyclohexanecarboxylic acid (MCCA) were examined in bile-exteriorized rats. A 0.52 mmol/kg dose (equivalent to 75 mg/kg VPA) of test compound (N = 4 rats per compound) was administered as an intravenous bolus. VPA, CCA, and MCCA concentrations in serum, bile, and urine were determined by gas chromatography before and after incubation in sodium hydroxide to hydrolyze base-labile conjugates. Concentration-time profiles of these compounds in serum displayed apparent Michaelis-Menten kinetics. Serum concentrations of base-labile conjugates were similar to parent concentrations for VPA, were an order of magnitude lower than parent concentrations for CCA, and were undetectable for MCCA. Urinary recovery of base-labile (apparently glucuronide) conjugates in the bile-exteriorized rat was 28.8%, 12.0%, and 25.2% of the administered dose for VPA, CCA, and MCCA, respectively. In contrast, more than 50% of the dose for VPA and MCCA was recovered in bile as the base-labile conjugate, with less than 5% of the CCA dose recovered via this excretory route. Bile flow was stimulated significantly by VPA and MCCA, but not by CCA; changes in bile flow correlated with the biliary excretion rate of base-labile conjugates rather than with excretion of the parent compounds themselves.(ABSTRACT TRUNCATED AT 250 WORDS)


A prospective evaluation of the Cape triage score in the emergency department of an urban public hospital in South Africa.
Until recently South Africa had no triage system for emergency department (ED) use. The Cape triage group developed a triage scale called the Cape triage score (CTS). This system consists of a basic physiology score, mobility score and a short list of important discriminators that cannot be accurately triaged on a physiological score alone. Highest priority is given to a red colour code, followed by orange, yellow and green. The purpose was to evaluate the components of the CTS and identify amendments that would improve the quality of the scale in terms of its accuracy to identify patients more likely to require admission or at high risk of death in the ED. Data were prospectively collected over a 4-month period. Data captured included the parameters of a basic physiological score (respiratory rate, pulse rate, systolic blood pressure, temperature and a simplified score measuring level of consciousness), mobility, a list of selected clinical conditions (discriminator list), final clinical diagnosis and final outcome in the ED (admission to hospital or death). 798 patients were triaged and analyzed. The CTS undertriaged 24% (overtriage 25%) of cases who required admission. By altering the colour code parameters, amending the discriminator list as well as the addition of a trauma factor, undertriage was reduced to 12% (with an overtriage of 45%). The amended CTS has an acceptably low undertriage rate and is capable of predicting patient disposal over a wide spectrum of ED presentations.


Trehalose facilitates DNA melting: a single-molecule optical tweezers study.
Using optical tweezers, here we show that the overstretching transition force of double-stranded DNA (dsDNA) is lowered significantly by the addition of the disaccharide trehalose as well as certain polyol osmolytes. This effect is found to depend linearly on the logarithm of the trehalose concentration. We propose an entropic driving mechanism for the experimentally observed destabilization of dsDNA that is rooted in the higher affinity of the DNA bases for trehalose than for water, which promotes base exposure and DNA melting. Molecular dynamics simulation reveals the direct interaction of trehalose with nucleobases. Experiments with other osmolytes confirm that the extent of dsDNA destabilization is governed by the ratio between polar and apolar fractions of an osmolyte.


Effect of single growth factor and growth factor combinations on differentiation of neural stem cells.
The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF), insulin growth factor-I (IGF-I), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using beta-tubulin III and NeuN as neural markers. Neural differentiations in the presence of individual growth factors for beta-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-1, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for beta-tubulin III and 3.1% for NeuN. For beta-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1%; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and postmitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.


Safer sex practices among newly diagnosed HIV-positive men who have sex with men in China: results from an ethnographic study.
The study reported here sought to understand the rationales of safer sex practices adopted by newly diagnosed HIV-positive men who have sex with men (MSM). Guided by a socio-ecological framework, an ethnography was conducted among newly diagnosed HIV-positive MSM. In-depth interviews and participant observation were employed to produce an account of the social and cultural settings that was faithful to the perspectives of participants. A total of 31 participants with diverse backgrounds were recruited in a southern city of China. Participant observation was conducted in local healthcare settings, MSM venues, and NGO offices. Most participants (24/31) reported stopping unprotected anal intercourse (UAI) immediately after being diagnosed as HIV-positive. Factors associated with safer sex practices were identified at both individual and environmental levels, including self-protection, establishment of self-esteem, dignity, altruism and reciprocity, disease experience as a source of personal growth, and organizational culture and values. Newly diagnosed HIV-positive MSM navigate their sexual practices within the context of multiple competing factors. Implications for sustained behaviour change enabling safer sex practices include stimulating survival instinct, facilitating safer sex decision making, motivating and facilitating personal growth, and encouraging volunteerism to promote intentional activities for safer sex practices.


Effects on mortality of alcohol consumption, smoking, physical activity, and close personal relationships.
The study analyses the risks of mortality associated with alcohol consumption and smoking, as well as possible counteracting effects of physical activity and social support through close personal relationships. Data are based on the Upper Bavarian Study, a longitudinal epidemiological study of a representative community sample (n = 1668) in a rural area. Extensive semistructured psychiatric interviews by research physicians were conducted between 1975 and 1977 (n = 1536). Thirteen years after psychiatric assessment, information was obtained from the community register concerning death in the interval, data of death and cause of death according to ICD 9. This information could be ascertained for 93.1% (n = 1430) of those who had been interviewed, thus providing a good basis for generalizing the findings. Results indicate that alcohol intake and cigarette smoking increased mortality while physical activity and the availability of a steady partner had protective effects. There were no interactive effects between the four variables studied, except for a dramatically increased risk for women drinking more than 20 ml of pure alcohol a day and reporting no physical exercise at wave one assessment. The relative risks of alcohol intake and smoking, and the counteracting effects of physical activity and partnership, are exemplified in the cases of a 40-year-old female and a 40-year-old male. Specific analyses of the relationship between alcohol consumption, smoking, physical exercise and personal relationships, on the one hand, and, on the other, different causes of death, are presented.


Meta-Analyses of the Effects of Habitual Running on Indices of Health in Physically Inactive Adults.
In order to implement running to promote physical activity, it is essential to quantify the extent to which running improves health. The aim was to summarise the literature on the effects of endurance running on biomedical indices of health in physically inactive adults. Electronic searches were conducted in October 2014 on PubMed, Embase, CINAHL, SPORTDiscus, PEDro, the Cochrane Library and LILACS, with no limits of date and language of publication. Randomised controlled trials (with a minimum of 8 weeks of running training) that included physically inactive but healthy adults (18-65 years) were selected. The studies needed to compare intervention (i.e. endurance running) and control (i.e., no intervention) groups. Two authors evaluated study eligibility, extracted data, and assessed risk of bias; a third author resolved any uncertainties. Random-effects meta-analyses were performed to summarise the estimates for length of training and sex. A dose-response analysis was performed with random-effects meta-regression in order to investigate the relationship between running characteristics and effect sizes. After screening 22,380 records, 49 articles were included, of which 35 were used to combine data on ten biomedical indices of health. On average the running programs were composed of 3.7 ± 0.9 sessions/week, 2.3 ± 1.0 h/week, 14.4 ± 5.4 km/week, at 60-90% of the maximum heart rate, and lasted 21.5 ± 16.8 weeks. After 1 year of training, running was effective in reducing body mass by 3.3 kg [95% confidence interval (CI) 4.1-2.5], body fat by 2.7% (95% CI 5.1-0.2), resting heart rate by 6.7 min(-1) (95% CI 10.3-3.0) and triglycerides by 16.9 mg dl(-1) (95% CI 28.1-5.6). Also, running significantly increased maximal oxygen uptake (VO2max) by 7.1 ml min(-1) kg(-1) (95% CI 5.0-9.1) and high-density lipoprotein (HDL) cholesterol by 3.3 mg dl(-1) (95% CI 1.2-5.4). No significant effect was found for lean body mass, body mass index, total cholesterol and low-density lipoprotein cholesterol after 1 year of training. In the dose-response analysis, larger effect sizes were found for longer length of training. It was only possible to combine the data of ten out the 161 outcome measures identified. Lack of information on training characteristics precluded a multivariate model in the dose-response analysis. Endurance running was effective in providing substantial beneficial effects on body mass, body fat, resting heart rate, VO2max, triglycerides and HDL cholesterol in physically inactive adults. The longer the length of training, the larger the achieved health benefits. Clinicians and health authorities can use this information to advise individuals to run, and to support policies towards investing in running programs.


Consumers' Perceptions of Edible Marijuana Products for Recreational Use: Likes, Dislikes, and Reasons for Use.
Edible marijuana products have become extremely popular in states that have legalized marijuana for recreational use. The goal of this research was to provide a better understanding of consumer perceptions of edible marijuana products, including why they prefer edibles relative to other forms of marijuana (e.g., smoking) and their concerns regarding the consumption of edibles. We conducted eight focus groups (four groups in Denver, Colorado, and four groups in Seattle, Washington) in February 2016 with 62 adult consumers of edibles. Focus group transcripts were coded in QSR NVivo 10.0 qualitative analysis software, and coding reports identified trends across participants. Most participants preferred edibles to smoking marijuana because there is no smell from smoke and no secondhand smoke. Other reasons participants like edibles included convenience, discreetness, longer-lasting highs, less intense highs, and edibles' ability to aid in relaxation and reduce anxiety more so than smoking marijuana. Concerns and dislikes about edibles included delayed effects, unexpected highs, the unpredictability of the high, and inconsistency of distribution of marijuana in the product. No participants in either location mentioned harmful health effects from consuming edibles as a concern. Conclusions/Importance: The present study was qualitative in nature and provides a good starting point for further research to quantify through surveys how consumers understand and use edibles. Such information will help guide policy makers and regulators as they establish regulations for edibles. Also, such research can help inform educational campaigns on proper use of edibles for recreational purposes.


Immunization with recombinant protein: conditions for cytotoxic T cell and/or antibody induction.
Safe vaccines should optimally induce both cell-mediated and humoral immunity. Recently, it has been shown that protective cytotoxic T cells (CTLs) can be induced not only with live vaccines, but also with recombinant viral proteins. This report shows in C57BL/6 (H-2b) mice that the recombinant nucleoprotein (N) of vesicular stomatitis virus (VSV) induced protective CTLs but no neutralizing antibodies in mice, whereas the recombinant glycoprotein (G) of VSV alone induced neutralizing antibodies but no CTLs. If the N and G of VSV were coinjected, both CTLs and a long-lasting neutralizing IgG response was measurable, demonstrating that mixed vaccines can be used to induce protective CTLs and antibodies with an efficiency comparable to live virus. In an attempt to define optimal conditions for CTL priming, the intravenous, intraperitoneal and subcutaneous route of injection were compared. Intravenous injection of recombinant VSV-N induced up to 30 times higher responses than the latter two routes. Finally, we tried to define conditions inducing only CTLs and no antibodies binding to the native protein form, or vice versa, only antibodies and no CTLs. Intravenous injection of boiled VSV-N induced a CTL response but no antibodies specific for the native VSV-N, whereas VSV-N injected subcutaneously in incomplete Freund's adjuvant induced high amounts of anti-VSV-N antibodies but virtually no CTLs. The conditions defined here permit vaccines to be designed which would function along selected and defined immunological effector pathways.


Provider Perspectives and Clinical Outcomes with Inpatient Telenephrology.
Despite the dramatic increase in the provision of virtual nephrology care, only anecdotal reports of outcomes without comparators to usual care exist in the literature. This study aimed to provide objective determination of clinical noninferiority of hybrid (telenephrology plus face-to-face) versus standard (face-to-face) inpatient nephrology care. This retrospective study compares objective outcomes in patients who received inpatient hybrid care versus standard nephrology care at two Mayo Clinic Health System community hospitals. Outcomes were then additionally compared with those patients receiving care at another Mayo Clinic Health System site where only standard care is available. Hospitalized adults who had nephrology consults from March 1, 2020 to February 28, 2021 were considered. Regression was used to assess 30-day mortality, length of hospitalization, readmissions, odds of being prescribed dialysis, and hospital transfers. Sensitivity analysis was performed using patients who had ≥50% of their care encounters via telenephrology. Structured surveys were used to understand the perspectives of non-nephrology hospital providers and telenephrologists. In total, 850 patients were included. Measured outcomes that included the number of hospital transfers (odds ratio, 1.19; 95% confidence interval, 0.37 to 3.82) and 30-day readmissions (odds ratio, 0.97; 95% confidence interval, 0.84 to 1.06), among others, did not differ significantly between controls and patients in the general cohort. Telenephrologists (n=11) preferred video consults (82%) to phone for communication. More than half (64%) of telenephrologists spent less time on telenephrology compared with standard care. Non-nephrology hospital providers (n=21) were very satisfied (48%) and satisfied (29%) with telenephrology response time and felt telenephrology was as safe as standard care (67%), while providing them enough information to make patient care decisions (76%). Outcomes for in-hospital nephrology consults were not significantly different comparing hybrid care versus standard care. Non-nephrology hospital providers and telenephrologists had favorable opinions of telenephrology and most perceived it is as safe and effective as standard care. This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2022_04_11_CJN13441021.mp3.


Synergistic effects of engineered nanoparticles and organics released from laser printers using nano-enabled toners: potential health implications from exposures to the emitted organic aerosol.
Recent studies have shown that engineered nanoparticles (ENPs) are incorporated into toner powder used in printing equipment and released during their use. Thus, understanding the functional and structural composition and potential synergistic effects of this complex aerosol and released gaseous co-pollutants is critical in assessing their potential toxicological implications and risks. In this study, toner powder and PEPs were thoroughly examined for functional and molecular composition of the organic fraction and the concentration profile of 16 Environmental Protection Agency (EPA)-priority polycyclic aromatic hydrocarbons (PAH) using state of the art analytical methods. Results show significant differences in abundance of non-exchangeable organic hydrogen of toner powder and PEPs, with a stronger aromatic spectral signature in PEPs. Changes in structural composition of PEPs are indicative of radical additions and free-radical polymerization favored by catalytic reactions, resulting in formation of functionalized organic species. Particularly, accumulation of aromatic carbons with strong styrene-like molecular signatures on PEPs is associated with formation of semivolatile heavier aromatic species (i.e., PAHs). Further, the transformation of low molecular weight PAHs in the toner powder to high molecular weight PAHs in PEPs was documented and quantified. This may be a result of synergistic effects from catalytic metal/metal oxide ENPs incorporated into the toner and the presence/release of semi-volatile organic species (SVOCs). The presence of known carcinogenic PAHs on PEPs raises public health concerns and warrants further toxicological assessment.


Recombinant bovine herpesvirus 4 (BoHV-4) expressing glycoprotein D of BoHV-1 is immunogenic and elicits serum-neutralizing antibodies against BoHV-1 in a rabbit model.
Several biological characteristics of bovine herpesvirus 4 (BoHV-4) make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity, unlikely oncogenicity, the capability to accommodate large amounts of foreign genetic material, the ability to infect several cell types coming from different animal species, and the ability to maintain transgene expression in both undifferentiated and differentiated cells. Starting from BoHV-4 cloned as a bacterial artificial chromosome (BAC), we used MuA transposase-mediated in vitro transposition to generate recombinant BoHV-4 expressing the immunodominant glycoprotein D (gD) of BoHV-1, one of the most important pathogens of cattle. Although a cis-acting element from woodchuck hepatitis virus (the woodchuck hepatitis virus posttranscriptional regulatory element [WPRE]) in the 3' end of the gD expression cassette was required for maximal gD expression from plasmids in transient transfection assays, this element was not necessary for efficient expression of gD from recombinant BoHV-4 genomes. BoHV-4 recombinants containing gD expression cassettes with or without the WPRE expressed gD at similarly high levels. Several cell lines originating from different animal species expressed gD when infected with BoHV-4 recombinants. When rabbits were immunized with one of the recombinants, high levels of serum neutralizing antibodies against BoHV-1 were generated. This work is one of the first demonstrations of the use BoHV-4 as a vector for vaccine purposes and may provide the basis for BoHV-1 vaccination of cattle with recombinant BoHV-4.


Primary Intestinal-type Adenocarcinoma of the Nasal Cavity: A Case Report.
We report a rare case of sinonasal intestinal-type adenocarcinoma in the nasal cavity. A 31-year old man represented with headache and epistaxis. We identified a malignant tumour, which is a rare pathology, with detailed physical examination, anterior rhinoscopy, computed tomography scan, magnetic resonance imaging and histopathologic examination. Endoscopic excision of the tumour was performed. After three years of follow-up of the patient in our clinic, there was no sign of any recurrence.


cDNA cloning and expression of rat homeobox gene, Hex, and functional characterization of the protein.
We isolated two cDNA clones of rat Hex, a homeobox protein, studied its expression in rat liver and various cells, and characterized the protein. The levels of Hex mRNA were only slightly increased in liver of rats refed with a high-carbohydrate diet or after partial hepatectomy. Whereas the expression of Hex mRNA was detected in hepatocytes isolated from adult rat liver and also in highly differentiated hepatoma cells, no Hex mRNA was detected in poorly differentiated hepatoma cells. Hex mRNA was also detected in liver from embryo aged 15 days. Expression of Hex was increased in F9 cells during differentiation into visceral endoderm cells by treatment with retinoic acid. This stimulation occurred prior to an increase in the level of alpha-fetoprotein mRNA. When fusion-protein expression vectors of GAL4 DNA-binding domain and Hex were co-transfected with luciferase reporter plasmid, with or without five copies of the GAL4-binding site, into HepG2 cells, the luciferase activities were decreased in concentration- and GAL4-binding site-dependent manners. This repression did not require the presence of the homeodomain, which is located between the amino acid residues 137 and 196. Its repression domain was mapped between the residues 45 and 136 in the proline-rich N-terminal region. In addition, the homeodomain was responsible for DNA-binding of Hex. These results indicate that Hex functions as a transcriptional repressor and may be involved in the differentiation and/or maintenance of the differentiated state in hepatocytes.


Myogenic potentials generated by a click-evoked vestibulocollic reflex.
Electromyograms (EMGs) were recorded from surface electrodes over the sternomastoid muscles and averaged in response to brief (0.1 ms) clicks played through headphones. In normal subjects, clicks 85 to 100 dB above our reference (45 dB SPL: close to perceptual threshold for normal subjects for such clicks) evoked reproducible changes in the averaged EMG beginning at a mean latency of 8.2 ms. The earliest potential change, a biphasic positive-negativity (p13-n23), occurred in all subjects and the response recorded from over the muscle on each side was predominantly generated by afferents originating from the ipsilateral ear. Later potentials (n34, p44), present in most but not all subjects, were generated bilaterally after unilateral ear stimulation. The amplitude of the averaged responses increased in direct proportion to the mean level of tonic muscle activation during the recording period. The p13-n23 response was abolished in patients who had undergone selective section of the vestibular nerve but was preserved in subjects with severe sensorineural hearing loss. It is proposed that the p13-n23 response is generated by activation of vestibular afferents, possibly those arising from the saccule, and transmitted via a rapidly conducting oligosynaptic pathway to anterior neck muscles. Conversely, the n34 and p44 potentials do not depend on the integrity of the vestibular nerve and probably originate from cochlear afferents.


THE INFLUENCE OF EPINEPHRIN UPON THE CORONARY CIRCULATION OF THE MONKEY.
Decrease in coronary flow was the constant response of freshly isolated monkey hearts to epinephrin. These hearts were perfused with autogenous hirudinized blood diluted with Locke solution. The results were constant at high or low perfusion pressures, in beating or resting hearts, and with all adequate doses. Increased coronary flow was obtained constantly in rabbit hearts under identical conditions. In the light of previous work upon isolated human coronary arteries, the general conclusion is drawn that, while actively dilating the coronary vessels in the dog, cat, rabbit, ox, sheep, and pig, epinephrin constricts the coronary vessels in man and the monkey. The coronary arteries of the last two species are presumably supplied with constrictor nerves of true sympathetic (thoracicolumbar) origin.


Aberrant expression of CD7 in myeloblasts is highly associated with de novo acute myeloid leukemias with FLT3/ITD mutation.
Acute myeloid leukemia (AML) with normal cytogenetics represents approximately 40% to 50% of de novo AML. This heterogeneous AML subgroup constitutes the single largest cytogenetic group with an intermediate prognosis. Previous studies have suggested that the Fms-like tyrosine kinase-3 internal tandem duplication (FLT3/ITD) mutation-positive de novo AML may represent a distinctive subgroup of AML. We analyzed the clinical and pathologic features of 15 cases of de novo AML with normal cytogenetics and with the FLT3/ITD mutation. In comparison with patients with AML without the FLT3/ITD mutation, patients with FLT3/ITD+ AML are relatively younger, more often have marked peripheral leukocytosis with a higher number of circulating blasts at initial examination, more often have minimal differentiation morphologic features, more frequently have abnormal CD7 coexpression, and have poorer outcome. Close association of aberrant CD7 expression and FLT3/ITD mutation in the myeloblasts of FLT3/ITD+ AML suggests that FLT3/ITD- mediated leukemic transformation occurs in the more early stage of myeloid progenitor cells.


An influenza prophylaxis clinic in a primary school: 24 hours from notification to protection.
A public health clinic was established to provide antiviral prophylaxis to school contacts during the pandemic (H1N1) 2009 influenza outbreak in NSW, Australia. Children (n = 74) and staff (n = 9) were provided with antiviral (oseltamivir) prophylaxis following exposure to a confirmed case of pandemic (H1N1) 2009 influenza. The success of the clinic included attention to infection control and quarantining of potentially infectious children and staff, attendance at the clinic of pharmacists to ensure accurate dispensing of suspension medication, availability of experienced public health staff at short notice, and provision of accurate information to staff, school children and families attending the clinic.


Electronic and Structural Effects of Inner Sphere Coordination of Chloride to a Homoleptic Copper(II) Diimine Complex.
The reaction of CuCl2 with 2,9-dimethyl-1,10-phenanthroline (dmp) does not lead to the formation of [Cu(dmp)2](Cl)2 but instead to [Cu(dmp)2Cl]Cl, a 5-coordinated complex, in which one chloride is directly coordinated to the metal center. Attempts at removing the coordinated chloride by changing the counterion by metathesis were unsuccessful and resulted only in the exchange of the noncoordinated chloride, as confirmed from a crystal structure analysis. Complex [Cu(dmp)2Cl]PF6 exhibits a reversible cyclic voltammogram characterized by a significant peak splitting between the reductive and oxidative waves (0.85 and 0.60 V vs NHE, respectively), with a half-wave potential E1/2 = 0.73 V vs NHE. When reduced electrochemically, the complex does not convert into [Cu(dmp)2]+, as one may expect. Instead, [Cu(dmp)2]+ is isolated as a product when the reduction of [Cu(dmp)2Cl]PF6 is performed with l-ascorbic acid, as confirmed by electrochemistry, NMR spectroscopy, and diffractometry. [Cu(dmp)2]2+ complexes can be synthesized starting from Cu(II) salts with weakly and noncoordinating counterions, such as perchlorate. Growth of [Cu(dmp)2](ClO4)2 crystals in acetonitrile results in a 5-coordinated complex, [Cu(dmp)2(CH3CN)](ClO4)2, in which a solvent molecule is coordinated to the metal center. However, solvent coordination is associated with a dynamic decoordination-coordination behavior upon reduction and oxidation. Hence, the cyclic voltammogram of [Cu(dmp)2(CH3CN)]2+ is identical to the one of [Cu(dmp)2]+, if the measurements are performed in acetonitrile. The current results show that halide ions in precursors to Cu(II) metal-organic coordination compound synthesis, and most likely also other multivalent coordination centers, are not readily exchanged when exposed to presumed strongly binding and chelating ligand, and thus special care needs to be taken with respect to product characterization.


Predicting outcome of assertive outreach across England.
Assertive community treatment for the severely mentally ill is being implemented increasingly internationally. It is unclear whether recommended characteristics of assertive outreach (AO) teams influence care and outcomes. We hypothesised that recommended characteristics of AO teams such as joint health and social care management would predict reduced hospitalisation in the first year of an AO client programme and related outcomes throughout England. A two-stage design was used: a stratified sample of 100 of the 186 'stand-alone' AO teams in England and a systematic sample of clients from each team with stratification for black and ethnic minority patients. Team characteristics, treatment and outcomes were collected from teams. Analyses took account of patients' histories, clustering and ethnic minority over-sampling. Under AO the proportion of time spent in hospital following admission decreased. Only 3/1,096 patients went missing in 9 months. Although patient' histories significantly predicted outcomes almost no team characteristics predicted re-admission or other patient outcomes after 1 and 3 years. Ethnic minority clients were more likely to be on compulsory orders only on jointly managed teams (P = 0.030). Multidisciplinary teams and teams not working out of hours significantly predicted that patients received psychological interventions, but only 17% of sampled patients received such treatments. Characteristics of AO teams do not explain long-term patient outcomes. Since recommended team characteristics are not effective new models of care should be developed and the process of care tested. Managing teams to implement evidence-based psychological interventions might improve outcomes.


Identification of IRF-8 and IRF-1 target genes in activated macrophages.
Interferon regulatory factor 1 (IRF-1) and IRF-8, also known as interferon consensus sequence binding protein (ICSBP), are important regulators of macrophage differentiation and function. These factors exert their activities through the formation of heterocomplexes. As such, they are coactivators of various interferon-inducible genes in macrophages. To gain better insights into the involvement of these two transcription factors in the onset of the innate immune response and to identify their regulatory network in activated macrophages, DNA microarray was employed. Changes in the expression profile were analyzed in peritoneal macrophages from wild type mice and compared to IRF-1 and IRF-8 null mice, before and following 4 h exposure to IFN-gamma and LPS. The expression pattern of 265 genes was significantly changed (up/down) in peritoneal macrophages extracted from wild type mice following treatment with IFN-gamma and LPS, while no changes in the expression levels of these genes were observed in samples of the same cell-type from both IRF-1 and IRF-8 null mice. Among these putative target genes, numerous genes are involved in macrophage activity during inflammation. The expression profile of 10 of them was further examined by quantitative RT-PCR. In addition, the promoter regions of three of the identified genes were analyzed by reporter gene assay for the ability to respond to IRF-1 and IRF-8. Together, our results suggest that both IRF-1 and IRF-8 are involved in the transcriptional regulation of these genes. We therefore suggest a broader role for IRF-1 and IRF-8 in macrophages differentiation and maturation, being important inflammatory mediators.


Trial of the cysteine cathepsin inhibitor JPM-OEt on early and advanced mammary cancer stages in the MMTV-PyMT-transgenic mouse model.
Recent data suggest proteases of the papain-like cysteine cathepsin family as molecular targets for cancer therapy. Here, we report the treatment of polyoma middle T oncogene-induced breast cancers in mice with the cell-permeable broad-spectrum cysteine cathepsin inhibitor JPM-OEt. Up to 100 mg/kg inhibitor was intraperitoneally injected once per day in two trials on early and advanced cancers. In both trials, transient delays in tumour growth were observed. However, at the endpoint of both experiments no significant differences in tumour weights, histopathology and lung metastasis were found between the inhibitor and the control group. The invasive strand formation of collagen I-embedded tumour cell spheroids generated from primary tumours of inhibitor-treated mice in the early cancer trial could be inhibited in vitro by JPM-OEt; a result arguing against induction of resistance to the inhibitor. Measurement of cysteine cathepsin activities in tissue extracts after intraperitoneal injection of JPM-OEt revealed effective inhibition of cysteine cathepsins in pancreas, kidneys and liver, while activities in mammary cancers and in lungs were not significantly affected. We conclude that the pharmacokinetic properties of JPM-OEt, which result in poor bioavailability, may prohibit its use for stand-alone treatment of solid mammary cancers and their lung metastases.


Ribosomal PCR and DNA sequencing for detection and identification of bacteria: experience from 6 years of routine analyses of patient samples.
The use of broad range PCR and DNA sequencing of bacterial 16S ribosomal RNA genes for routine diagnostics of bacterial infections was evaluated. Here, the results from more than 2600 analyses during a 6-year period (2003-2009) are presented. Almost half of the samples were from joints and bones, and the second most frequent origin of samples was from the central nervous system. Overall, 26% of all samples were positive for bacterial DNA and bacterial identification was obtained in 80% of the PCR-positive samples by subsequent DNA sequencing. Ambiguous species identification was noticed among non-haemolytic streptococci, especially within the mitis group. The data show that ribosomal PCR with subsequent DNA sequencing of the PCR product is a most valuable supplement to culture for identifying bacterial agents of both acute and prolonged infections. However, some bacteria, including non-haemolytic streptococci, may not be precisely identified.


A transgenic model of visceral obesity and the metabolic syndrome.
The adverse metabolic consequences of obesity are best predicted by the quantity of visceral fat. Excess glucocorticoids produce visceral obesity and diabetes, but circulating glucocorticoid levels are normal in typical obesity. Glucocorticoids can be produced locally from inactive 11-keto forms through the enzyme 11beta hydroxysteroid dehydrogenase type 1 (11beta HSD-1). We created transgenic mice overexpressing 11beta HSD-1 selectively in adipose tissue to an extent similar to that found in adipose tissue from obese humans. These mice had increased adipose levels of corticosterone and developed visceral obesity that was exaggerated by a high-fat diet. The mice also exhibited pronounced insulin-resistant diabetes, hyperlipidemia, and, surprisingly, hyperphagia despite hyperleptinemia. Increased adipocyte 11beta HSD-1 activity may be a common molecular etiology for visceral obesity and the metabolic syndrome.


Tsx produces a long noncoding RNA and has general functions in the germline, stem cells, and brain.
The Tsx gene resides at the X-inactivation center and is thought to encode a protein expressed in testis, but its function has remained mysterious. Given its proximity to noncoding genes that regulate X-inactivation, here we characterize Tsx and determine its function in mice. We find that Tsx is actually noncoding and the long transcript is expressed robustly in meiotic germ cells, embryonic stem cells, and brain. Targeted deletion of Tsx generates viable offspring and X-inactivation is only mildly affected in embryonic stem cells. However, mutant embryonic stem cells are severely growth-retarded, differentiate poorly, and show elevated cell death. Furthermore, male mice have smaller testes resulting from pachytene-specific apoptosis and a maternal-specific effect results in slightly smaller litters. Intriguingly, male mice lacking Tsx are less fearful and have measurably enhanced hippocampal short-term memory. Combined, our study indicates that Tsx performs general functions in multiple cell types and links the noncoding locus to stem and germ cell development, learning, and behavior in mammals.


Geometric symmetry of the solar surface of hooves of thoroughbred racehorses.
To define a 3-dimensional (3-D) coordinate system with clear definitions of origins and axes relative to hoof anatomic features and determine whether solar surfaces of Thoroughbred racehorse hooves have geometric asymmetry in the mediolateral and dorsopalmar directions. Left forelimb hooves from 20 Thoroughbred racehorse cadavers. A right-handed 3-D coordinate axes system centered on the collateral sulci was defined for the left front hoof. Orthogonal distances of anatomic features from the dorsopalmar axis and the plane coincident with the ground were measured and compared between medial and lateral sides and between dorsal and palmar regions of the hoof. The hoof was wider and had a greater radius laterally than medially. The most distal part of the lateral bar of the frog was further from the dorsopalmar axis than that of the medial bar. Overall, mediolateral asymmetries in depth were not observed. The sole at the perimeter was deeper medially in the dorsal part of the hoof and laterally in the palmar part, with depth overall being greater palmarly than dorsally. Most features had dorsopalmar asymmetry. When the angle bisected by the collateral sulci is used to determine the dorsopalmar axis of the hoof, most central structures (bars and collateral sulci) have mediolateral symmetry. However, the hoof wall and sole have some mediolateral asymmetries and most structures have dorsopalmar asymmetry. These findings may assist the development of devices for attachment to hooves and studies of the interaction of hooves with bearing surfaces.


Determination of PD-L1 expression in effusions from mesothelioma by immuno-cytochemical staining.
Malignant mesothelioma (MM) is an aggressive, fatal tumor. Current therapeutic options only marginally improve survival. Programmed cell death ligand 1 (PD-L1) is a dominant mediator of immunosuppression, binding to programmed cell death 1 (PD-1). PD-L1 is up-regulated in cancer cells, and the PD-1/PD-L1 pathway plays a critical role in tumor immune evasion, thus providing a target for antitumor therapy. Further, a correlation between PD-L1 expression and prognosis has been reported. Studies performed on histological material have revealed expression of PD-L1 in MM, but no study has been performed on MM effusions thus far. PD-L1 expression was determined by a commercially available antibody (clone 28-8) in 74 formalin-fixed, paraffin-embedded cell blocks from body effusions obtained at diagnosis from patients with MM. The presence of MM cells was confirmed with CK5/6, calretinin, and EMA and the admixture of macrophages was assessed with CD68. Only cases containing more than 100 tumor cells were assessed. Membranous staining in tumor cells was considered positive. Survival time was calculated from the appearance of the first malignant effusion until death. Reactivity was observed in 23 of 61 (38%) of cases and was classified as ≥1%-5% (n = 9 cases), >5%-10% (n = 4 cases), >10%-50% (n = 4 cases), and >50% (n = 6 cases) positive cells. Survival times did not differ significantly between patients with PD-L1-positive and PD-L1-negative tumors. MM effusions are suitable for immune-cytochemical assessment of PD-L1 expression in malignant cells and the results are similar to those reported for histological specimens. Cancer Cytopathol 2017;125:908-17. © 2017 American Cancer Society.


SU-E-I-60: Validation of An Optically Stimulated Luminescent (OSL) Dosimeter for Use in Output Exposure Control Verification of Mammography Imaging Systems.
To develop and validate an Optically Stimulated Luminescent (OSL) dosimeter for exposure control verification of x-ray projection mammography imaging systems. The active detection element of the dosimeter is a strip of OSL material 3.0 mm wide, 0.13 mm thick and 30.0 mm long with an overlying aluminum step wedge with thicknesses of 0, 0.2, 0.4 and 0.6 mm Al, encapsulated in a light-tight plastic enclosure with outer dimensions of 10.0 mm wide, 5.4 mm thick, and 54.0 mm long. The dosimeter is used in conjunction with a breast phantom for the purpose of estimating the half-value layer (HVL), entrance surface exposure (ESE), and average glandular dose (AGD) in conventional projection mammography. ESE and HVL were computed based on analysis of exposure profiles obtained from exposed strip dosimeters. The AGD was estimated by multiplying the ESE by the appropriate exposure to dose conversion factor for the thickness and % glandular tissue fraction represented by the phantom and target-filter combination employed. The accuracy and reproducibility of the ESE, HVL and AGD estimates obtained using the dosimeter positioned on the surface of the ACR phantom at the chest wall edge, was evaluated using mammography systems utilizing different imaging receptor technology, i.e. screen-film (SF), computed radiography (CR) and direct radiography (DR) and compared against results obtained using a calibrated ion chamber fitted with a mammography probe. ESE, AGD and HVL results obtained using the OSL mammography QA dosimeter agreed with results obtained using an ion chamber to within 5-10%, depending on the target-filter combination used. Repeat readings were highly consistent with a coefficient of variation = 5%. The OSL mammography QA dosimeter has been shown to effectively estimate ESE, HVL and AGD, demonstrating its usefulness for secondary monitoring of output exposure of mammography imaging systems.


CollaboNet: collaboration of deep neural networks for biomedical named entity recognition.
Finding biomedical named entities is one of the most essential tasks in biomedical text mining. Recently, deep learning-based approaches have been applied to biomedical named entity recognition (BioNER) and showed promising results. However, as deep learning approaches need an abundant amount of training data, a lack of data can hinder performance. BioNER datasets are scarce resources and each dataset covers only a small subset of entity types. Furthermore, many bio entities are polysemous, which is one of the major obstacles in named entity recognition. To address the lack of data and the entity type misclassification problem, we propose CollaboNet which utilizes a combination of multiple NER models. In CollaboNet, models trained on a different dataset are connected to each other so that a target model obtains information from other collaborator models to reduce false positives. Every model is an expert on their target entity type and takes turns serving as a target and a collaborator model during training time. The experimental results show that CollaboNet can be used to greatly reduce the number of false positives and misclassified entities including polysemous words. CollaboNet achieved state-of-the-art performance in terms of precision, recall and F1 score. We demonstrated the benefits of combining multiple models for BioNER. Our model has successfully reduced the number of misclassified entities and improved the performance by leveraging multiple datasets annotated for different entity types. Given the state-of-the-art performance of our model, we believe that CollaboNet can improve the accuracy of downstream biomedical text mining applications such as bio-entity relation extraction.


Healthy lungs tolerate repetitive collapse and reopening during short periods of mechanical ventilation.
The possible occurrence of lung damage if alveolar units are allowed to collapse and reopen breath by breath during mechanical ventilation with normal tidal volumes was investigated. Anaesthetised, paralysed, open chest rabbits were subjected to either intrathoracic negative (NEEP; n = 6) or positive (PEEP; n = 6) end-expiratory pressure during volume controlled mechanical ventilation. Both experimental settings were preceded by a 30 min control period and followed by a 30 min recovery period during which a PEEP of 0.2 kPa was maintained. Pao2 and pulmonary compliance deteriorated significantly in the NEEP group during the experimental period and compared to ventilation with PEEP. Partial restoration of lung mechanics and blood gases was achieved during the recovery period. After an alveolar recruitment manoeuvre, this recovery was complete. Lung clearance studied by depositing an aerosol of technetium-99m-labelled diethylenetriamine pentaacetate (99mTc-DTPA) in the alveoli, was significantly faster during ventilation with NEEP compared to the PEEP group (P = 0.0002) as well as the control period (P = 0.0029). It did not recover completely during the recovery period but remained significantly faster. Light microscopic histology was normal in both groups with no evidence of inflammation or epithelial disruption. We conclude that previously healthy rabbit lungs show only a transient disturbance of lung mechanics and blood gases with repetitive collapse and re-expansion. The integrity of the alveolar microstructure is preserved. The disturbance in the alveolo-capillary permeability persists and may indicate surfactant related alveolo-capillary barrier dysfunction.


The validity of the Annual Review of Competence Progression: a qualitative interview study of the perceptions of junior doctors and their trainers.
Objective To investigate trainee doctors' and trainers' perceptions of the validity of the Annual Review of Competence Progression (ARCP) using Messick's conceptualisation of construct validity. Design Qualitative semi-structured focus groups and interviews with trainees and trainers. Setting Postgraduate medical training in London, Kent Surrey and Sussex, Yorkshire and Humber, and Wales in November/December 2015. Part of a larger study about the fairness of postgraduate medical training. Participants Ninety-six trainees and 41 trainers, comprising UK and international medical graduates from Foundation, General Practice, Medicine, Obstetrics and Gynaecology, Psychiatry, Radiology, and Surgery, at all levels of training. Main outcome measures Trainee and trainer perceptions of the validity of the ARCP as an assessment tool. Results Participants recognised the need for assessment, but were generally dissatisfied with ARCPs, especially UK graduate trainees. Participants criticised the perceived tick-box nature of ARCPs as measuring clerical rather than clinical ability, and which they found detrimental to learning. Trainees described being able to populate their e-portfolios with just positive feedback; they also experienced difficulty getting assessments signed off by supervisors. ARCPs were perceived as poor at identifying struggling trainees and/or as discouraging excellence by focussing on minimal competency. Positive experiences of ARCPs arose when trainees could discuss their progress with interested supervisors. Conclusions Trainee and trainer criticisms of ARCPs can be conceptualised as evidence that ARCPs lack validity as an assessment tool. Ongoing reforms to workplace-based assessments could address negative perceptions of the 'tick-box' elements, encourage constructive input from seniors and allow trainees to demonstrate excellence as well as minimal competency, while keeping patients safe.


Vitamin D in a northern Canadian first nation population: dietary intake, serum concentrations and functional gene polymorphisms.
The wide spectrum of vitamin D activity has focused attention on its potential role in the elevated burden of disease in a northern Canadian First Nations (Dené) cohort. Vitamin D insufficiency, and gene polymorphisms in the vitamin D receptor (VDR) and vitamin D binding protein (VDBP) have been implicated in susceptibility to infectious and chronic diseases. The objectives of this study were to determine the contribution of vitamin D from food, and measure the serum concentrations of 25-hydroxyvitamin D(3) (25-OHD(3)) and VDBP in Dené participants. Single nucleotide polymorphisms (SNPs) associated with the dysregulation of the innate immune response were typed and counted. Potential correlations between the SNPs and serum concentrations of 25-OHD(3) and VDBP were evaluated. Venous blood was collected in summer and winter over a one-year period and analyzed for 25-OHD(3) and VDBP concentrations (N = 46). A questionnaire was administered to determine the amount of dietary vitamin D consumed. Sixty-one percent and 30% of the participants had 25-OHD(3) serum concentrations <75 nmol/L in the winter and summer respectively. Mean vitamin D binding protein concentrations were within the normal range in the winter but below normal in the summer. VDBP and VDR gene polymorphisms affect the bioavailability and regulation of 25-OHD(3). The Dené had a high frequency of the VDBP D432E-G allele (71%) and the Gc1 genotype (90%), associated with high concentrations of VDBP and a high binding affinity to 25-OHD(3). The Dené had a high frequency of VDR Fok1-f allele (82%), which has been associated with a down-regulated Th1 immune response. VDBP and VDR polymorphisms, and low winter 25-OHD(3) serum concentrations may be risk factors for infectious diseases and chronic conditions related to the dysregulation of the vitamin D pathway.


Effects of neonicotinoids and fipronil on non-target invertebrates.
We assessed the state of knowledge regarding the effects of large-scale pollution with neonicotinoid insecticides and fipronil on non-target invertebrate species of terrestrial, freshwater and marine environments. A large section of the assessment is dedicated to the state of knowledge on sublethal effects on honeybees (Apis mellifera) because this important pollinator is the most studied non-target invertebrate species. Lepidoptera (butterflies and moths), Lumbricidae (earthworms), Apoidae sensu lato (bumblebees, solitary bees) and the section "other invertebrates" review available studies on the other terrestrial species. The sections on freshwater and marine species are rather short as little is known so far about the impact of neonicotinoid insecticides and fipronil on the diverse invertebrate fauna of these widely exposed habitats. For terrestrial and aquatic invertebrate species, the known effects of neonicotinoid pesticides and fipronil are described ranging from organismal toxicology and behavioural effects to population-level effects. For earthworms, freshwater and marine species, the relation of findings to regulatory risk assessment is described. Neonicotinoid insecticides exhibit very high toxicity to a wide range of invertebrates, particularly insects, and field-realistic exposure is likely to result in both lethal and a broad range of important sublethal impacts. There is a major knowledge gap regarding impacts on the grand majority of invertebrates, many of which perform essential roles enabling healthy ecosystem functioning. The data on the few non-target species on which field tests have been performed are limited by major flaws in the outdated test protocols. Despite large knowledge gaps and uncertainties, enough knowledge exists to conclude that existing levels of pollution with neonicotinoids and fipronil resulting from presently authorized uses frequently exceed the lowest observed adverse effect concentrations and are thus likely to have large-scale and wide ranging negative biological and ecological impacts on a wide range of non-target invertebrates in terrestrial, aquatic, marine and benthic habitats.


Novel reprogramming of neutrophils modulates inflammation resolution during atherosclerosis.
Nonresolving inflammation perpetuated by innate leukocytes is involved in the pathogenesis of unstable atherosclerosis. However, the role and regulation of neutrophils related to nonresolving inflammation and atherosclerosis are poorly understood. We report herein that chronic subclinical endotoxemia, a risk factor for atherosclerosis, skewed neutrophils into a nonresolving inflammatory state with elevated levels of inflammatory mediators (Dectin-1, MMP9, and LTB4) and reduced levels of homeostatic mediators (LRRC32, TGFβ, and FPN). The polarization of neutrophils was due to ROS-mediated activation of oxCAMKII, caused by altered peroxisome homeostasis and reduced lysosome fusion. Application of 4-phenylbutyrate (4-PBA) enhanced peroxisome homeostasis of neutrophils, reduced oxCAMKII, and rebalanced the expression profiles of pro- and anti-inflammatory mediators. Adoptive transfer of neutrophils programmed by subclinical endotoxemia rendered exacerbated atherosclerosis. In contrast, transfer of ex vivo programmed neutrophils by 4-PBA reduced the pathogenesis of atherosclerosis. Our data define novel neutrophil dynamics associated with the progression and regression of atherosclerosis.


Differential effects of habitat loss on occupancy patterns of the eastern green lizard Lacerta viridis at the core and periphery of its distribution range.
The effects of habitat loss on the distribution of populations are often linked with species specialization degree. Specialist species can be more affected by changes in landscape structure and local patch characteristics compared to generalist species. Moreover, the spatial scale at which different land covers (eg. habitat, cropland, urban areas) affect specialist species can be smaller. Specialization is usually assumed as a constant trait along the distribution range of species. However, for several taxa, there is evidence of higher specialization degree in peripheral populations compared with populations in the core. Hence, peripheral populations should have a higher sensitivity to habitat loss, and strongest effects should be found at a smaller spatial scale. To test these expectations, we implemented a patch-landscape approach at different spatial scales, and compared effects of landscape structure and patch characteristics on occupancy probability among northern peripheral, more specialized populations (Czech Republic) and core populations (Bulgaria) of the eastern green lizard Lacerta viridis. We found that landscape structure and patch characteristics affect differently the occupancy probability of Lacerta viridis in each region. Strongest effects of habitat loss were found at a spatial scale of 150m around patches in the periphery, but at a scale of 500m in the core. In the periphery occupancy probability of populations was principally affected by landscape composition, and the effect of habitat quality was stronger compared to core populations. In the core, persistence of populations was mainly explained by characteristics of the spatial configuration of habitat patches. We discuss possible ecological mechanisms behind the relationship between sensitivity to habitat loss, populations' specialization degree and position in the distribution range, and suggest conservation measures for L. viridis.


The relative performance of Bayesian and parsimony approaches when sampling characters evolving under homogeneous and heterogeneous sets of parameters.
We tested whether it is beneficial for the accuracy of phylogenetic inference to sample characters that are evolving under different sets of parameters, using both Bayesian MCMC (Markov chain Monte Carlo) and parsimony approaches. We examined differential rates of evolution among characters, differential character-state frequencies and character-state space, and differential relative branch lengths among characters. We also compared the relative performance of parsimony and Bayesian analyses by progressively incorporating more of these heterogeneous parameters and progressively increasing the severity of this heterogeneity. Bayesian analyses performed better than parsimony when heterogeneous simulation parameters were incorporated into the substitution model. However, parsimony outperformed Bayesian MCMC when heterogeneous simulation parameters were not incorporated into the Bayesian substitution model. The higher the rate of evolution simulated, the better parsimony performed relative to Bayesian analyses. Bayesian and parsimony analyses converged in their performance as the number of simulated heterogeneous model parameters increased. Up to a point, rate heterogeneity among sites was generally advantageous for phylogenetic inference using both approaches. In contrast, branch-length heterogeneity was generally disadvantageous for phylogenetic inference using both parsimony and Bayesian approaches. Parsimony was found to be more conservative than Bayesian analyses, in that it resolved fewer incorrect clades.


Analysis of 28 generations of selection for reproduction, growth, and carcass traits in swine.
Selection (28 generations, G) in a Large White-Landrace composite population for traits aimed at increasing live pigs born per litter (BA), with additional selection for increased 180-d weight (WT180) and longissimus muscle area (LMA) and decreased back fat (BF10) in the last 8 generations, was practiced. Objectives herein were to estimate genetic and phenotypic responses and genetic parameters (n = 1,883 to 54,174) and to investigate whether a plateau in response for BA occurred. Line 2 (L2) was selected for an index of ovulation rate and embryo survival (G0 to G11), fully formed pigs (FF) per litter (G12 to 14), and BA and pig birth weight (PBW, G15 to G19), and its control line (LC1) was selected randomly (G0 to G21). Line 4 (L4), derived from L2, and line 5 (L5), derived from LC1, at G8 were selected in 2 stages for ovulation rate and FF (G9 to G16) and BA and PBW (G17 to G19), and their control (LC6) was selected randomly. At G20, L4 and L5 were crossed to form L45, and L4 and L2 were crossed to continue L2; L2 and L45 were subsequently selected for BA, WT180, LMA, and BF10 (G21 to G28). At G21, LC1 and LC6 were reciprocally crossed to form LC16, control for L2, and LC61, control for L45. Selection in L2 and L45 was first for BA and then for other traits among pigs selected for BA. Line sizes were 40 to 60 litters by 15 to 20 sires/G. Cumulative selection differentials (CSD) were calculated. MTDFREML was used to estimate variance components, EBV, and responses. Genetic changes at G28 in L2 were 4.63 FF and 3.66 BA, with 72% (FF) and 86% (BA) of the change occurring after G11. Two-stage selection produced similar responses (P < 0.01) in FF in L4 and L5 (0.27 and 0.29 pigs/G) but a greater response in BA in L5 (0.19 vs. 0.28 pigs/G). Genetic change in L45 from G20 to G28 was 0.17 pigs/G for both FF and BA (P < 0.01). Genetic changes at G28 in L45 were 4.16 FF and 3.68 BA. Genetic correlations of reproductive and growth traits were near zero, ranging from -0.43 (stillborn pigs/litter with BF10) to 0.21 (mummies/litter with LMA). Selection for growth traits along with litter size selection during G19 to G28 resulted in responses consistent with the selection applied and the heritability of the trait. No evidence for a selection plateau existed; selection differentials and variances of FF and BA in selection lines during G20 to G28 were similar to those in earlier generations. Over all generations, heritability of BA was 0.20 ± 0.03 and remained at approximately 0.17 in selection lines in later generations.