Reporting on blinding in trial protocols and corresponding publications was often inadequate but rarely contradictory.
To compare the reporting on blinding in protocols and articles describing randomized controlled trials. We studied 73 protocols of trials approved by the scientific/ethical committees for Copenhagen and Frederiksberg, 1994 and 1995, and their corresponding publications. Three out of 73 trials (4%) reported blinding in the protocol that contradicted that in the publication (e.g., "open" vs. "double blind"). The proportion of "double-blind" trials with a clear description of the blinding of participants increased from 11 out of 58 (19%) when based on publications alone to 39 (67%) when adding the information in the protocol. The similar proportions for the blinding of health care providers were 2 (3%) and 22 (38%); and for the blinding of data collectors, they were 8 (14%) and 14 (24%). In 52 of 58 publications (90%), it was unclear whether all patients, health care providers, and data collectors had been blinded. In 4 of the 52 trials (7%), the protocols clarified that all three key trial persons had been blinded. The reporting on blinding in both trial protocols and publications is often inadequate. We suggest developing international guidelines for the reporting of trial protocols and public access to protocols.


Dietary zinc and the control of Streptococcus pneumoniae infection.
Human zinc deficiency increases susceptibility to bacterial infection. Although zinc supplementation therapies can reduce the impact of disease, the molecular basis for protection remains unclear. Streptococcus pneumoniae is a major cause of bacterial pneumonia, which is prevalent in regions of zinc deficiency. We report that dietary zinc levels dictate the outcome of S. pneumoniae infection in a murine model. Dietary zinc restriction impacts murine tissue zinc levels with distribution post-infection altered, and S. pneumoniae virulence and infection enhanced. Although the activation and infiltration of murine phagocytic cells was not affected by zinc restriction, their efficacy of bacterial control was compromised. S. pneumoniae was shown to be highly sensitive to zinc intoxication, with this process impaired in zinc restricted mice and isolated phagocytic cells. Collectively, these data show how dietary zinc deficiency increases sensitivity to S. pneumoniae infection while revealing a role for zinc as a component of host antimicrobial defences.


NK cell function and antibodies mediating ADCC in HIV-1-infected viremic and controller patients.
Natural killer (NK) cells have been suggested to play a protective role in HIV disease progression. One potent effector mechanism of NK cells is antibody-dependent cellular cytotoxicity (ADCC) mediated by antiviral antibodies binding to the FcγRIIIa receptor (CD16) on NK cells. We investigated NK cell-mediated ADCC function and the presence of ADCC antibodies in plasma from 20 HIV-1-infected patients and 10 healthy donors. The HIV-positive patients were divided into two groups: six who controlled viremia for at least 8 y without treatment (controllers), and 14 who were persistently viremic and not currently on treatment. Plasma from both patient groups induced NK cell IFN-γ expression and degranulation in response to HIV-1 envelope (Env) gp140-protein-coated cells. Patient antibodies mediating ADCC were largely directed towards the Env V3 loop, as identified by a gp140 protein lacking the V3 loop. Interestingly, in two controllers ADCC-mediating antibodies were more broadly directed to other parts of Env. A high viral load in patients correlated with decreased ADCC-mediated cytolysis of gp140-protein-coated target cells. NK cells from both infected patients and healthy donors degranulated efficiently in the presence of antibody-coated HIV-1-infected Jurkat cells. In conclusion, the character of ADCC-mediating antibodies differed in some controllers compared to viremic patients. NK cell ADCC activity is not compromised in HIV-infected patients.


Radiological response predicts survival following transarterial chemoembolisation in patients with unresectable hepatocellular carcinoma.
It remains unclear whether initial compact lipiodol uptake after transarterial chemoembolisation (TACE) is associated with improved survival in patients with hepatocellular carcinoma (HCC). To reveal the clinical relevance of compact lipiodolisation after TACE. We studied 490 patients with unresectable HCC who had first been treated with TACE. Compact lipiodolisation was defined as the absence of an arterial enhancing lesion, reflecting complete lipiodol uptake, as assessed by dynamic computed tomography (CT) 1 month after treatment. The rate of initial compact lipiodolisation was analysed according to multiplicity and size of tumour, and survival of patients who achieved compact lipiodolisation was compared to that of patients who did not. Of the 490 patients, 409 (83.5%) were in Child-Pugh class A and 81 (16.5%) in class B. The rate of initial compact lipiodolisation in single HCCs was higher than that in multinodular HCCs (33.7% vs. 14.6%, P < 0.001). Among single HCCs, the rate of compact lipiodolisation in tumours ≤5, 5-10 and >10 cm was 46.6%, 13.6%, and 0% respectively. The 1-, 3- and 5-year survival rates of patients with compact uptake were 92.7%, 70.7% and 52.4% compared to 60.8%, 28.0% and 16.9% in patients with noncompact lipiodolisation. Multivariate analysis revealed that Child-Pugh class, alpha-fetoprotein level, tumour node metastasis stage, portal vein thrombosis and initial compact lipiodolisation were independent predictors of survival. Initial compact lipiodol uptake after transarterial chemoembolisation is associated with improved survival in patients with unresectable hepatocellular carcinoma. Accordingly, initial complete lipiodolisation should be considered a relevant therapeutic target.


Characterizing spatial uncertainty when integrating social data in conservation planning.
Recent conservation planning studies have presented approaches for integrating spatially referenced social (SRS) data with a view to improving the feasibility of conservation action. We reviewed the growing conservation literature on SRS data, focusing on elicited or stated preferences derived through social survey methods such as choice experiments and public participation geographic information systems. Elicited SRS data includes the spatial distribution of willingness to sell, willingness to pay, willingness to act, and assessments of social and cultural values. We developed a typology for assessing elicited SRS data uncertainty which describes how social survey uncertainty propagates when projected spatially and the importance of accounting for spatial uncertainty such as scale effects and data quality. These uncertainties will propagate when elicited SRS data is integrated with biophysical data for conservation planning and may have important consequences for assessing the feasibility of conservation actions. To explore this issue further, we conducted a systematic review of the elicited SRS data literature. We found that social survey uncertainty was commonly tested for, but that these uncertainties were ignored when projected spatially. Based on these results we developed a framework which will help researchers and practitioners estimate social survey uncertainty and use these quantitative estimates to systematically address uncertainty within an analysis. This is important when using SRS data in conservation applications because decisions need to be made irrespective of data quality and well characterized uncertainty can be incorporated into decision theoretic approaches.


The impact of violence on Venezuelan life expectancy and lifespan inequality.
Venezuela is one of the most violent countries in the world. According to the United Nations, homicide rates in the country increased from 32.9 to 61.9 per 100 000 people between 2000 and 2014. This upsurge coincided with a slowdown in life expectancy improvements. We estimate mortality trends and quantify the impact of violence-related deaths and other causes of death on life expectancy and lifespan inequality in Venezuela. Life tables were computed with corrected age-specific mortality rates from 1996 to 2013. From these, changes in life expectancy and lifespan inequality were decomposed by age and cause of death using a continuous-change model. Lifespan inequality, or variation in age at death, is measured by the standard deviation of the age-at-death distribution. From 1996 to 2013 in Venezuela, female life expectancy rose 3.57 [95% confidence interval (CI): 3.08-4.09] years [from 75.79 (75.98-76.10) to 79.36 (78.97-79.68)], and lifespan inequality fell 1.03 (-2.96 to 1.26) years [from 18.44 (18.01-19.00) to 17.41 (17.30-18.27)]. Male life expectancy increased 1.64 (1.09-2.25) years [from 69.36 (68.89-59.70) to 71.00 (70.53-71.39)], but lifespan inequality increased 0.95 (-0.80 to 2.89) years [from 20.70 (20.24-21.08) to 21.65 (21.34-22.12)]. If violence-related death rates had not risen over this period, male life expectancy would have increased an additional 1.55 years, and lifespan inequality would have declined slightly (-0.31 years). As increases in violence-related deaths among young men (ages 15-39) have slowed gains in male life expectancy and increased lifespan inequality, Venezuelan males face more uncertainty about their age at death. There is an urgent need for more accurate mortality estimates in Venezuela.


Population based allele frequencies of disease associated polymorphisms in the Personalized Medicine Research Project.
There is a lack of knowledge regarding the frequency of disease associated polymorphisms in populations and population attributable risk for many populations remains unknown. Factors that could affect the association of the allele with disease, either positively or negatively, such as race, ethnicity, and gender, may not be possible to determine without population based allele frequencies.Here we used a panel of 51 polymorphisms previously associated with at least one disease and determined the allele frequencies within the entire Personalized Medicine Research Project population based cohort. We compared these allele frequencies to those in dbSNP and other data sources stratified by race. Differences in allele frequencies between self reported race, region of origin, and sex were determined. There were 19544 individuals who self reported a single racial category, 19027 or (97.4%) self reported white Caucasian, and 11205 (57.3%) individuals were female. Of the 11,208 (57%) individuals with an identifiable region of origin 8337 or (74.4%) were German.41 polymorphisms were significantly different between self reported race at the 0.05 level. Stratification of our Caucasian population by self reported region of origin revealed 19 polymorphisms that were significantly different (p = 0.05) between individuals of different origins. Further stratification of the population by gender revealed few significant differences in allele frequencies between the genders. This represents one of the largest population based allele frequency studies to date. Stratification by self reported race and region of origin revealed wide differences in allele frequencies not only by race but also by region of origin within a single racial group. We report allele frequencies for our Asian/Hmong and American Indian populations; these two minority groups are not typically selected for population allele frequency detection. Population wide allele frequencies are important for the design and implementation of studies and for determining the relevance of a disease associated polymorphism for a given population.


What can cross-cultural correlations teach us about human nature?
Many recent evolutionary psychology and human behavioral ecology studies have tested hypotheses by examining correlations between variables measured at a group level (e.g., state, country, continent). In such analyses, variables collected for each aggregation are often taken to be representative of the individuals present within them, and relationships between such variables are presumed to reflect individual-level processes. There are multiple reasons to exercise caution when doing so, including: (1) the ecological fallacy, whereby relationships observed at the aggregate level do not accurately represent individual-level processes; (2) non-independence of data points, which violates assumptions of the inferential techniques used in null hypothesis testing; and (3) cross-cultural non-equivalence of measurement (differences in construct validity between groups). We provide examples of how each of these gives rise to problems in the context of testing evolutionary hypotheses about human behavior, and we offer some suggestions for future research.


Clinical outcomes with ertapenem as a first-line treatment option of infections caused by extended-spectrum β-lactamase producing gram-negative bacteria.
Infections caused by extended-spectrum β-lactamase (ESBL)-producing gram-negative organisms are a growing concern in hospitalized patients. Traditionally, these infections can be effectively treated by the carbapenem class of drugs. In 2005, our institution initiated a protocol for use of ertapenem, a carbapenem, as the first-line treatment option for these infections. It is unknown whether ertapenem is associated with similar clinical response and microbiologic cure rates as those achieved with group 2 carbapenems (imipenem, meropenem, doripenem). To describe clinical response and microbiologic cure rates associated with ertapenem as first-line treatment of infections caused by ESBL-producing organisms. This case series included patients who received ertapenem for more than 48 hours to treat a documented infection with a positive culture for an ESBL-producing organism. Efficacy was determined by the clinical response and microbiologic cure rates achieved with ertapenem. Seventy-three patients received ertapenem for a mean (SD) of 10.7 (5.9) days. The most common (59%) infection site was urine. The most common causative organisms were ESBL-producing Klebsiella pneumoniae (47%) and Escherichia coli (48%). Clinical response was observed in 78% of patients. Microbiologic cure was achieved in 92% of the evaluable population (n = 50). There were no significant differences in clinical or microbiologic cure rates across important subgroups. Patients treated with ertapenem achieved favorable clinical response and microbiologic cure rates. Our data suggest that ertapenem can be used as an alternative to group 2 carbapenems for the treatment of infections caused by ESBL-producing gram-negative organisms.


Dynamic expression of the Robo ligand Slit2 in bone marrow cell populations.
The bone marrow (BM) niche is essential for lifelong hematopoietic stem cell (HSC) maintenance, proliferation and differentiation. Several BM cell types, including osteoblast lineage cells (OBC), mesenchymal stem cells (MSC) and endothelial cells (EC) have been implicated in supporting HSC location and function, but the relative importance of these cell types and their secreted ligands remain controversial. We recently found that the cell surface receptors Robo4 and CXCR4 cooperate to localize HSC to BM niches. We hypothesized that Slit2, a putative ligand for Robo4, cooperates with the CXCR4 ligand SDF1 to direct HSC to specific BM niche sites. Here, we have isolated OBC, MSC and EC by flow cytometry and determined their frequency within the bone marrow and the relative mRNA levels of Slit2, SDF1 and Robo4. We found that expression of Slit2 and SDF1 were dynamically regulated in MSC and OBC-like populations following radiation, while Robo4 expression was restricted to EC. Radiation also significantly affected the cellularity and frequency of both the non-adherent and adherent cells within the BM stroma. These data support a physiological role for Slit2 in regulating the dynamic function of Robo-expressing cells within BM niches at steady state and following radiation.


Sensitivity of the reflection technique: optimum angles of incidence to determine the optical properties of materials.
The purpose of this study is to develop a technique that will provide the maximum sensitivity of the reflection technique for the determination of the optical properties of materials. Analytical relations are presented that yield the maximum sensitivity of the technique for a given range of angles of incidence and refractive indices. The methods of the reflectance ratio at one and two angles of incidence are compared. The angular range of 75 degrees to 80 degrees was found to be suitable for the maximum sensitivity for the reflectance ratio at one angle of incidence, whereas for the two angles of incidence the ranges 45 degrees to 60 degrees and 75 degrees to 85 degrees are suitable. Furthermore, angular-reflectance measurements on a specular carbon-rod surface at the wavelength of 3.5 microm were used to demonstrate the advantages of inverting full-angularrange data as opposed to one or two angle measurements. The limitations of the measurements at normal incidence and 45 degrees are also assessed.


Mutations in alpha-tubulin cause abnormal neuronal migration in mice and lissencephaly in humans.
The development of the mammalian brain is dependent on extensive neuronal migration. Mutations in mice and humans that affect neuronal migration result in abnormal lamination of brain structures with associated behavioral deficits. Here, we report the identification of a hyperactive N-ethyl-N-nitrosourea (ENU)-induced mouse mutant with abnormalities in the laminar architecture of the hippocampus and cortex, accompanied by impaired neuronal migration. We show that the causative mutation lies in the guanosine triphosphate (GTP) binding pocket of alpha-1 tubulin (Tuba1) and affects tubulin heterodimer formation. Phenotypic similarity with existing mouse models of lissencephaly led us to screen a cohort of patients with developmental brain anomalies. We identified two patients with de novo mutations in TUBA3, the human homolog of Tuba1. This study demonstrates the utility of ENU mutagenesis in the mouse as a means to discover the basis of human neurodevelopmental disorders.


Ocean acidification induces distinct transcriptomic responses across life history stages of the sea urchin Heliocidaris erythrogramma.
Ocean acidification (OA) from seawater uptake of rising carbon dioxide emissions impairs development in marine invertebrates, particularly in calcifying species. Plasticity in gene expression is thought to mediate many of these physiological effects, but how these responses change across life history stages remains unclear. The abbreviated lecithotrophic development of the sea urchin Heliocidaris erythrogramma provides a valuable opportunity to analyse gene expression responses across a wide range of life history stages, including the benthic, post-metamorphic juvenile. We measured the transcriptional response to OA in H. erythrogramma at three stages of the life cycle (embryo, larva, and juvenile) in a controlled breeding design. The results reveal a broad range of strikingly stage-specific impacts of OA on transcription, including changes in the number and identity of affected genes; the magnitude, sign, and variance of their expression response; and the developmental trajectory of expression. The impact of OA on transcription was notably modest in relation to gene expression changes during unperturbed development and much smaller than genetic contributions from parentage. The latter result suggests that natural populations may provide an extensive genetic reservoir of resilience to OA. Taken together, these results highlight the complexity of the molecular response to OA, its substantial life history stage specificity, and the importance of contextualizing the transcriptional response to pH stress in light of normal development and standing genetic variation to better understand the capacity for marine invertebrates to adapt to OA.


Testing in the HIV Counselling and Testing (HCT) campaign, HIV risk behaviours and ART enrolment in South Africa.
To examine the association between testing in the 2010 HIV Testing and Counselling (HCT) campaign with HIV risk behaviours and enrolment on ART. Data for this study were collected as part of a nationally representative cross-sectional household survey conducted in 2012 in South Africa. Consenting participants completed a structured questionnaire and provided a dry blood spot specimen which was tested for HIV antibodies and antiretroviral drugs. Multinomial logistic regression was used to examine the association between HIV testing history and explanatory variables of interest. There was no association between testing in the 2010 HCT campaign and condom use at last sex, number of sexual partnerships or HIV knowledge. Individuals who tested in the HCT campaign were more likely to disclose their status (COR 2.6, 95% CI: 1.71-3.8) and those who tested HIV positive in the campaign were more likely to be receiving ART (COR 1.8, 95% CI: 1.1-2.9). Testing in the HCT campaign was associated with having received both pretest and post-test counselling while testing before the campaign was associated with having received HIV results with no counselling (COR 2.1, 95% CI: 1.2-3.8). We highlight the success of the 2010 HCT campaign in improving HIV status disclosure and enrolment on ART as well as shortcomings on HIV risk behaviours and HIV knowledge. These may be related to issues of quality assurance in the counselling process. Our results further highlight possible HCT counselling inconsistencies across sectors requiring stronger public-private partnership in the delivery of HCT in South Africa.


Risk factors associated with repetition of self-harm in black and minority ethnic (BME) groups: a multi-centre cohort study.
Little information is available to inform clinical assessments on risk of self-harm repetition in ethnic minority groups. In a prospective cohort study, using data collected from six hospitals in England for self-harm presentations occurring between 2000 and 2007, we investigated risk factors for repeat self-harm in South Asian and Black people in comparison to Whites. During the study period, 751 South Asian, 468 Black and 15,705 White people presented with self-harm in the study centres. Repeat self-harm occurred in 4379 individuals, which included 229 suicides (with eight of these fatalities being in the ethnic minority groups). The risk ratios for repetition in the South Asian and Black groups compared to the White group were 0.6, 95% CI 0.5-0.7 and 0.7, 95% CI 0.5-0.8, respectively. Risk factors for repetition were similar across all three groups, although excess risk versus Whites was seen in Black people presenting with mental health symptoms, and South Asian people reporting alcohol use and not having a partner. Additional modelling of repeat self-harm count data showed that alcohol misuse was especially strongly linked with multiple repetitions in both BME groups. Ethnicity was not recorded in a third of cases which may introduce selection bias. Differences may exist due to cultural diversity within the broad ethnic groups. Known social and psychological features that infer risk were present in South Asian and Black people who repeated self-harm. Clinical assessment in these ethnic groups should ensure recognition and treatment of mental illness and alcohol misuse.


Compounds with Therapeutic Potential against Novel Respiratory 2019 Coronavirus.
Currently, the expansion of the novel human respiratory coronavirus (known as SARS-CoV-2 [severe acute respiratory syndrome coronavirus 2], COVID-2019 [coronavirus disease 2019], or 2019-nCoV [2019 novel coronavirus]) has stressed the need for therapeutic alternatives to alleviate and stop this new epidemic. The previous epidemics of infections by high-morbidity human coronaviruses, such as SARS-CoV in 2003 and the Middle East respiratory syndrome coronavirus (MERS-CoV) in 2012, prompted the characterization of compounds that could be potentially active against the currently emerging novel coronavirus, SARS-CoV-2. The most promising compound is remdesivir (GS-5734), a nucleotide analog prodrug currently in clinical trials for treating Ebola virus infections. Remdesivir inhibited the replication of SARS-CoV and MERS-CoV in tissue cultures, and it displayed efficacy in nonhuman animal models. In addition, a combination of the human immunodeficiency virus type 1 (HIV-1) protease inhibitors lopinavir/ritonavir and interferon beta (LPV/RTV-IFN-β) was shown to be effective in patients infected with SARS-CoV. LPV/RTV-IFN-β also improved clinical parameters in marmosets and mice infected with MERS-CoV. Remarkably, the therapeutic efficacy of remdesivir appeared to be superior to that of LPV/RTV-IFN-β against MERS-CoV in a transgenic humanized mouse model. The relatively high mortality rates associated with these three novel human coronavirus infections, SARS-CoV, MERS-CoV, and SARS-CoV-2, have suggested that proinflammatory responses might play a role in the pathogenesis. It remains unknown whether the generated inflammatory state should be targeted. Therapeutics that target the coronavirus alone might not be able to reverse highly pathogenic infections. This minireview aims to provide a summary of therapeutic compounds that have shown potential in fighting SARS-CoV-2 infections.


Does Metacognition Make a Unique Contribution to Health Anxiety When Controlling for Neuroticism, Illness Cognition, and Somatosensory Amplification?
Somatosensory amplification (e.g., Barsky, 1992), illness cognition (e.g., Salkovskis & Warwick, 1986), and neuroticism (e.g., Noyes et al., 2003) have all been linked to health anxiety. The first two factors are disorder specific; however, neuroticism is a general vulnerability connected to a range of disorders. In the metacognitive model (Wells, 2009), beliefs about thinking have been implicated in the development of psychopathologies, but little is known about the contribution of individual differences in metacognition to health anxiety, specifically. A cross-sectional design was employed with convenience sampling used for participant selection. Participants (N = 351) completed a questionnaire battery and the following hypotheses were tested: (a) metacognition will show a significant positive correlation with health anxiety and (b) the relationship between metacognition and health anxiety will remain significant after controlling for variables normally associated with health anxiety (i.e., neuroticism, somatosensory amplification, and illness cognition). Hierarchical multiple regression analysis were run to test hypotheses and determine the best independent metacognitive predictors. The results supported each of the hypotheses and revealed three independent metacognitive predictors of health anxiety: "negative metacognitive beliefs about uncontrollability and danger," "beliefs about the need for thought control," and "cognitive confidence." Overall, this study indicates that metacognition may have an important role in health anxiety, and the clinical implications are discussed.


Thoracic outlet syndrome--a myofascial variant: Part 3. Structural and postural considerations.
Thoracic outlet syndrome involves more than just local neurovascular compression. Myofascial release treatments and stretching exercises may be only partially or temporarily successful unless all related components of somatic dysfunction, including craniosacral mechanisms, are addressed. Structural and postural abnormalities in the frontal plane, as with a short leg, and in the sagittal plane, such as lumbopelvic imbalances, as well as neural involvement all contribute to thoracic outlet syndrome symptoms. Once segmental restrictions are treated and symptoms diminish, postural correction and strengthening exercises may be initiated. Osteopathic diagnosis and treatment of the local, regional, and remote structural problems is necessary for optimal treatment of thoracic outlet syndrome and the maintenance of a symptom-free status.


Incidence and variance of knee injuries in elite college football players.
Knee injuries are among the most common musculoskeletal injuries in US football players. The literature includes little information about the role of player position and risk for knee injury. We hypothesized that the incidence of knee injury in elite collegiate US football players is high and that type of injury varies by player position. We evaluated 332 elite collegiate US football players at the 2005 National Football League Combine. All players underwent radiographic examinations, including plain x-rays and/or magnetic resonance imaging when necessary. All knee pathologic conditions and surgical procedures were recorded. Data were analyzed by player position to detect any trends. Fifty-four percent (179) of the 332 players had a history of knee injury; knee injuries totaled 233 (1.3/player injured). Eighty-six players (25.9%) had a total of 114 surgeries. The most common injuries were medial collateral ligament injury (n = 79), meniscal injury (n = 51), and anterior cruciate ligament (ACL) injury (n = 40). The most common surgeries were arthroscopic meniscectomy (n = 39), ACL reconstruction (n = 35), and arthroscopic meniscal repair (n = 13). A history of knee injury was most common in defensive linemen (68% of players), tight ends (57%), and offensive linemen (57%). Knee surgery was more commonly performed on running backs (36%) and linebackers (34%). There were no significant associations between type or frequency of specific injuries with regard to player position. Knee injuries are common injuries in elite collegiate football players, and one fourth of these players undergo surgical procedures. However, there were no statistically significant differences in type or frequency of injuries by player position.


Diversity and productivity peak at intermediate dispersal rate in evolving metacommunities.
Positive relationships between species diversity and productivity have been reported for a number of ecosystems. Theoretical and experimental studies have attempted to determine the mechanisms that generate this pattern over short timescales, but little attention has been given to the problem of understanding how diversity and productivity are linked over evolutionary timescales. Here, we investigate the role of dispersal in determining both diversity and productivity over evolutionary timescales, using experimental metacommunities of the bacterium Pseudomonas fluorescens assembled by divergent natural selection. We show that both regional diversity and productivity peak at an intermediate dispersal rate. Moreover, we demonstrate that these two patterns are linked: selection at intermediate rates of dispersal leads to high niche differentiation between genotypes, allowing greater coverage of the heterogeneous environment and a higher regional productivity. We argue that processes that operate over both ecological and evolutionary timescales should be jointly considered when attempting to understand the emergence of ecosystem-level properties such as diversity-function relationships.


Power supplies and equipment for military field research: lessons from the British Service Dhaulagiri Research Expedition 2016.
The British Service Dhaulagiri Research Expedition (BSDMRE) took place from 27 March to 31 May 2016. The expedition involved 129 personnel, with voluntary participation in nine different study protocols. Studies were conducted in three research camps established at 3600, 4600 and 5140 m and involved taking and storing blood samples, cardiac echocardiography and investigations involving a balance plate. Research in this remote environment requires careful planning in order to provide a robust and resilient power plan. In this paper we aim to report the rationale for the choices we made in terms of power supply, the equipment used and potential military applicability. This is a descriptive account from the expedition members involved in planning and conducting the medical research. Power calculations were used to determine estimates of requirement prior to the expedition. The primary sources used to generate power were internal combustion engine (via petrol fuelled electric generators) and solar panels. Having been generated, power was stored using lithium-ion batteries. Special consideration was given to the storage of samples taken in the field, for which electric freezers and dry shippers were used. All equipment used functioned well during the expedition, with the challenges of altitude, temperature and transport all overcome due to extensive prior planning. Power was successfully generated, stored and delivered during the BSDMRE, allowing extensive medical research to be undertaken. The challenges faced and overcome are directly applicable to delivering military medical care in austere environments, and lessons learnt can help with the planning and delivery of future operations, training exercises or expeditions.


The exploration of risk factors of concurrent bacteraemia in patients critically ill with severe dengue.
We investigated the clinical features of intensive care unit (ICU) patients with concomitant severe dengue infection and bacteraemia to identify risk factors for this comorbidity. The records of all ICU dengue patients admitted during the period of 31 July-30 November 2015 were reviewed. Patients with 'concurrent bacteremia' (positive bacterial blood culture within 72 h of ICU admission) were identified. ICU admission was required for 142 patients, of which 22 (15.5 %) had concurrent bacteraemia. Species of the genus Streptococcus was the most common pathogens, followed by Escherichia coli then species of the genus Staphylococcus. Patients with a severe dengue infection and bacteraemia had higher APACHE II and TISS scores, C-reactive protein (CRP) levels and leukocyte counts, positive fluid balances, longer activated partial thromboplastin times (APTTs), higher lactate levels and more kidney failure, but controls (severe dengue patients without bacteraemia) had higher Glasgow Coma Scale (GCS) scores, higher albumin levels and more abdominal pain (all P<0.05). Patients with bacteraemia had a higher mortality rate than did ontrols (40.9 vs 18.3 %; P=0.018). Multiple logistic regression analysis showed that bacteraemia was significantly positively associated with the following independent predictors: higher CRP levels [adjusted odds ratio (aOR): 1.026; 95 % confidence interval (CI): 1.008-1.044; P=0.005], and longer APTTs (aOR: 1.034; 95 CI: 1.004-1.065; P=0.027). Concurrent bacteraemia is not uncommon in severe dengue patients in the ICU, and it is associated with high mortality. Higher CRP levels and longer APTTs were two independent risk factors associated with bacteraemia.


The stimulation of insulin secretion in non-insulin-dependent diabetic patients by amino acids and gliclazide in the basal and hyperglycemic state.
Sulfonylureas stimulate insulin secretion as their predominant contribution toward decreasing blood glucose in diabetic patients. We studied eight gliclazide-treated, non-insulin-dependent diabetic patients on two occasions with a protocol of basal observation for 30 minutes, a 60-minute infusion of randomized leucine or arginine, and a further 90-minute hyperglycemic clamp. Basal glucose was the same on both occasions (mean, 7.82 mmol/L for leucine v 7.79 for arginine, P = NS), and glucose levels declined to 7.50 and 7.25 mmol/L, respectively, by 30 minutes. After leucine infusion, the decline of glucose continued, but stabilized or reversed with arginine such that by the end of the infusions, glucose levels were 6.63 +/- 0.69 mmol/L for leucine and 7.62 +/- 0.67 for arginine (P < .02). Arginine caused a sharp increase in insulin secretion (from 17.8 mU/L to 43.8 mU/L in 6 minutes) at the onset of the infusion, and thereafter insulin secretion was not significantly different throughout either the amino acid or hyperglycemic clamp periods (mean, 42.1 v 44.7 mU/L, respectively, P = NS). By contrast, the leucine infusion caused little acute change in secretion, but augmented it with time from the basal period (17.2 mU/L) to the end of the infusion (29.4 mU/L). During the hyperglycemic clamp period, there was significant further augmentation of insulin secretion, increasing to 81.6 +/- 16 mU/L at the end of the study. Leucine significantly augmented insulin secretion compared with arginine (81.6 +/- 16 v 54.0 +/- 8.4 mU/L, respectively, P < .002). These data suggest that leucine is a better priming agent for sulfonylurea than arginine. Additive effects on insulin secretion may allow the use of combinations of branched chain amino acids (BCAAs) and sulfonylureas to augment insulin secretion in the presence of hyperglycemia.


Double in situ hybridization techniques in zebrafish.
Over recent years many genes expressed in zebrafish embryos have been cloned and a first step in their analysis is the determination of the spatial and temporal expression patterns of their transcripts by in situ hybridization. It is often necessary to relate the expression pattern to that of other genes expressed at the same period of development. Transcripts from different genes may be expressed in complementary or overlapping domains. The pattern of expression of different genes can be related by performing in situ hybridization on consecutive sections of tissue but it is also possible to perform hybridizations with two probes and to visualize the signals separately in the same tissue. Occasionally it is also possible to combine in situ hybridization with immunocytochemistry to localize tissue-specific antigens. Methods developed for performing these types of analyses in zebrafish are described in this article.


Luminal preloading with hydrogen-rich saline ameliorates ischemia-reperfusion injury following intestinal transplantation in rats.
Prolonged intestinal cold storage causes considerable mucosal breakdown, which could bolster bacterial translocation and cause life-threatening infection for the transplant recipient. The intestine has an intraluminal compartment, which could be a target for intervention, but has not yet been fully investigated. Hydrogen gas exerts organ protection and has used been recently in several clinical and basic research studies on topics including intestinal transplantation. In this study, we aimed to investigate the cytoprotective efficacy of intraluminally administered hydrogen-rich saline on cold IR injury in intestinal transplantation. Isogeneic intestinal transplantation with 6 hours of cold ischemia was performed on Lewis rats. Hydrogen-rich saline (H2 concentration at 5 ppm) or normal saline was intraluminally introduced immediately before preservation. Graft intestine was excised 3 hours after reperfusion and analyzed. Histopathological analysis of control grafts revealed blunting of the villi and erosion. These mucosal changes were notably attenuated by intraluminal hydrogen. Intestinal mucosa damage caused by IR injury led to considerable deterioration of gut barrier function 3 h post-reperfusion. However, this decline in permeability was critically prevented by hydrogen treatment. IR-induced upregulation of proinflammatory cytokine mRNAs such as IL-6 was mitigated by hydrogen treatment. Western blot revealed that hydrogen treatment regulated loss of the transmembrane protein ZO-1. Hydrogen-rich saline intraluminally administered in the graft intestine modulated IR injury to transplanted intestine in rats. Successful abrogation of intestinal IR injury with a novel strategy using intraluminal hydrogen may be easily clinically applicable and will compellingly improve patient care after transplantation.


Interstitial deletion of 2q associated with craniosynostosis, ocular coloboma, and limb abnormalities: cytogenetic and molecular investigation.
We report on the clinical and cytogenetic findings in a 9-year-old boy with a de novo deletion of 2q, shown by molecular analysis to have arisen from the paternal chromosome. Examination of microsatellite markers indicated deletion of bands 2q24.3 and 2q31. Clinical findings included craniosynostosis, bilateral ocular colobomata, and limb abnormalities, the latter being an emerging association with deletion of this region of 2q.


Prefrontal Cortex Oxygenation Evoked by Convergence Load Under Conflicting Stimulus-to-Accommodation and Stimulus-to-Vergence Eye-Movements Measured by NIRS.
Purpose: To extend our knowledge of the functional linkages between visual fatigue and regional cerebral prefrontal cortex (PFC) oxygenation, we measured time related hemodynamic changes over the right dorsolateral prefrontal cortex (dlPFC) during convergence load under conflicting stimulus-to-accommodation and stimulus-to-vergence eye movements with and without concurrent mental load. Methods: Twenty healthy participants with a median age of 28 years (range: 18-44 years) fixated upon a vertical bar presented separately to the left and right eyes, using polarized filters, during four counterbalanced 10-min periods: (i) no accommodation/vergence conflict (Control, Ctrl); (ii) added convergence load and accommodation/vergence conflict (Conv); (iii) added cognitive load only (Cog) and; (iv) a combination of added cognitive and convergence load and accommodation/vergence conflict (Cc). Viewing distance was 65 cm. Non-invasive measurements of hemodynamic activity over the dlPFC were quantified by functional near-infrared spectroscopy (fNIRS). During the two-convergence load conditions, the horizontal disparity of the two bars varied dynamically from no disparity to a disparity set 20% below the individual threshold for diplopia. Cognitive load was induced by the n-back-2 test which required the subject to memorize and recall the changing colors of the horizontal bars and decide when a given color was the same as that occurring two colors previously. fNIRS data were averaged over 10-s windows centered at 0, 2, 4, 6, 8, and 10 min of each task, subtracted from a 20-s baseline window immediately preceding the visual task, and then represented as changes in oxygenated hemoglobin (ΔHbO2), deoxygenated hemoglobin (ΔHHb) and total hemoglobin (ΔtHb). Results: Linear mixed model analyses showed that hemodynamic activity was systematically influenced by time (p < 0.001). The group-averaged time-related level of change across the viewing conditions did not differ when compared with one another (p > 0.05). Larger convergence eye-movement responses under conflicting stimulus-to-accommodation, and stimulus-to-vergence over time, increased ΔHbO2 and ΔtHb only in condition Cc and after 8 min of task time (p < 0.10 for min-6 and min-8: p < 0.05 for min-10). Discussion: Collectively, our data suggest that HbO2, HHb, and tHb, recorded over the dlPFC with fNIRS, can be used to assay the degree to which supervisory oculomotor control processes are activated during visually deficient near work.


The role of photon scattering in voltage-calcium fluorescent recordings of ventricular fibrillation.
Recent optical mapping studies of cardiac tissue suggest that membrane voltage (V(m)) and intracellular calcium concentrations (Ca) become dissociated during ventricular fibrillation (VF), generating a proarrhythmic substrate. However, experimental methods used in these studies may accentuate measured dissociation due to differences in fluorescent emission wavelengths of optical voltage/calcium (V(opt)/Ca(opt)) signals. Here, we simulate dual voltage-calcium optical mapping experiments using a monodomain-Luo-Rudy ventricular-tissue model coupled to a photon-diffusion model. Dissociation of both electrical, V(m)/Ca, and optical, V(opt)/Ca(opt), signals is quantified by calculating mutual information (MI) for VF and rapid pacing protocols. We find that photon scattering decreases MI of V(opt)/Ca(opt) signals by 23% compared to unscattered V(m)/Ca signals during VF. Scattering effects are amplified by increasing wavelength separation between fluorescent voltage/calcium signals and respective measurement-location misalignment. In contrast, photon scattering does not affect MI during rapid pacing, but high calcium dye affinity can decrease MI by attenuating alternans in Ca(opt) but not in V(opt). We conclude that some dissociation exists between voltage and calcium at the cellular level during VF, but MI differences are amplified by current optical mapping methods.


Calbindin and parvalbumin are early markers of non-mitotically regenerating hair cells in the bullfrog vestibular otolith organs.
Earlier studies have demonstrated hair cell regeneration in the absence of cell proliferation, and suggested that supporting cells could phenotypically convert into hair cells following hair cell loss. Because calcium-binding proteins are involved in gene up-regulation, cell growth, and cell differentiation, we wished to determine if these proteins were up-regulated in scar formations and regenerating hair cells following gentamicin treatment. Calbindin and parvalbumin immunolabeling was examined in control or gentamicin-treated (GT) bullfrog saccular and utricular explants cultured for 3 days in amphibian culture medium or amphibian culture medium supplemented with aphidicolin, a blocker of nuclear DNA replication in eukaryotic cells. In control cultures, calbindin and parvalbumin immunolabeled the hair bundles and, less intensely, the cell bodies of mature hair cells. In GT or mitotically-blocked GT (MBGT) cultures, calbindin and parvalbumin immunolabeling was also seen in the hair bundles, cuticular plates, and cell bodies of hair cells with immature hair bundles. Thus, these antigens were useful markers for both normal and regenerating hair cells. Supporting cell immunolabeling was not seen in control cultures nor in the majority of supporting cells in GT cultures. In MBGT cultures, calbindin and parvalbumin immunolabeling was up-regulated in the cytosol of single supporting cells participating in scar formations and in supporting cells with hair cell-like characteristics. These data provide further evidence that non-mitotic hair cell regeneration in cultures can be accomplished by the conversion of supporting cells into hair cells.


Effects of infusion of human methemoglobin solution following hydrogen sulfide poisoning.
We have recently reported that infusion of a solution containing methemoglobin (MetHb) during exposure to hydrogen sulfide results in a rapid and large decrease in the concentration of the pool of soluble/diffusible H2S in the blood. However, since the pool of dissolved H2S disappears very quickly after H2S exposure, it is unclear if the ability of MetHb to "trap" sulfide in the blood has any clinical interest and relevance in the treatment of sulfide poisoning. In anesthetized rats, repetition of short bouts of high level of H2S infusions was applied to allow the rapid development of an oxygen deficit. A solution containing MetHb (600 mg/kg) or its vehicle was administered 1 min and a half after the end of H2S intoxication. The injection of MetHb solution increased methemoglobinemia to about 6%, almost instantly, but was unable to affect the blood concentration of soluble H2S, which had already vanished at the time of infusion, or to increase combined H2S. In addition, H2S-induced O2 deficit and lactate production as well as the recovery of carotid blood flow and blood pressure were similar in treated and control animals. Our results do not support the view that administration of MetHb or drugs-induced methemoglobinemia during the recovery phase following severe H2S intoxication in sedated rats can restore cellular oxidative metabolism, as the pool of diffusible sulfide, accessible to MetHb, disappears rapidly from the blood after H2S exposure.


FOXM1 expression is significantly associated with chemotherapy resistance and adverse prognosis in non-serous epithelial ovarian cancer patients.
Epithelial ovarian cancer (EOC) is a spectrum of different diseases, which makes their treatment a challenge. Forkhead box M1 (FOXM1) is an oncogene aberrantly expressed in many solid cancers including serous EOC, but its role in non-serous EOCs remains undefined. We examined FOXM1 expression and its correlation to prognosis across the three major EOC subtypes, and its role in tumorigenesis and chemo-resistance in vitro. Gene signatures were generated by microarray for 14 clear-cell and 26 endometrioid EOCs, and 15 normal endometrium snap-frozen biopsies. Validation of FOXM1 expression was performed by RT-qPCR and immunohistochemistry in the same samples and additionally in 50 high-grade serous EOCs and in their most adequate normal controls (10 luminal fallopian tube and 20 ovarian surface epithelial brushings). Correlations of FOXM1 expression to clinic-pathological parameters and patients' prognosis were evaluated by Kaplan-Meier and Cox proportional-hazards analyses. OVCAR-3 and two novel deeply characterized EOC cell lines (EOC-CC1 and OSPC2, with clear-cell and serous subtype, respectively) were employed for in vitro studies. Effects of FOXM1 inhibition by transient siRNA transfection were evaluated on cell-proliferation, cell-cycle, colony formation, invasion, and response to conventional first- and second-line anticancer agents, and to the PARP-inhibitor olaparib. Gene signatures of FOXM1-silenced cell lines were generated by microarray and confirmed by RT-qPCR. A significant FOXM1 mRNA up-regulation was found in EOCs compared to normal controls. FOXM1 protein overexpression significantly correlated to serous histology (p = 0.001) and advanced FIGO stage (p = 0.004). Multivariate analyses confirmed FOXM1 protein overexpression as an independent indicator of worse disease specific survival in non-serous EOCs, and of shorter time to progression in platinum-resistant cases. FOXM1 downregulation in EOC cell lines inhibited cell growth and clonogenicity, and promoted the cytotoxic effects of platinum compounds, doxorubicin hydrochloride and olaparib. Upon FOXM1 knock-down in EOC-CC1 and OSPC2 cells, microarray and RT-qPCR analyses revealed the deregulation of several common and other unique subtype-specific FOXM1 putative targets involved in cell cycle, metastasis, DNA repair and drug response. FOXM1 is up-regulated in all three major EOCs subtypes, and is a prognostic biomarker and a potential combinatorial therapeutic target in platinum resistant disease, irrespective of tumor histology.


Interim Results of a Multicenter Trial with the New Electronic Subretinal Implant Alpha AMS in 15 Patients Blind from Inherited Retinal Degenerations.
Purpose: We assessed the safety and efficacy of a technically advanced subretinal electronic implant, RETINA IMPLANT Alpha AMS, in end stage retinal degeneration in an interim analysis of two ongoing prospective clinical trials. The purpose of this article is to describe the interim functional results (efficacy). Methods: The subretinal visual prosthesis RETINA IMPLANT Alpha AMS (Retina Implant AG, Reutlingen, Germany) was implanted in 15 blind patients with hereditary retinal degenerations at four study sites with a follow-up period of 12 months (www.clinicaltrials.gov NCT01024803 and NCT02720640). Functional outcome measures included (1) screen-based standardized 2- or 4-alternative forced-choice (AFC) tests of light perception, light localization, grating detection (basic grating acuity (BaGA) test), and Landolt C-rings; (2) gray level discrimination; (3) performance during activities of daily living (ADL-table tasks). Results: Implant-mediated light perception was observed in 13/15 patients. During the observation period implant mediated localization of visual targets was possible in 13/15 patients. Correct grating detection was achieved for spatial frequencies of 0.1 cpd (cycles per degree) in 4/15; 0.33 cpd in 3/15; 0.66 cpd in 2/15; 1.0 cpd in 2/15 and 3.3 cpd in 1/15 patients. In two patients visual acuity (VA) assessed with Landolt C- rings was 20/546 and 20/1111. Of 6 possible gray levels on average 4.6 ± 0.8 (mean ± SD, n = 10) were discerned. Improvements (power ON vs. OFF) of ADL table tasks were measured in 13/15 patients. Overall, results were stable during the observation period. Serious adverse events (SAEs) were reported in 4 patients: 2 movements of the implant, readjusted in a second surgery; 4 conjunctival erosion/dehiscence, successfully treated; 1 pain event around the coil, successfully treated; 1 partial reduction of silicone oil tamponade leading to distorted vision (silicon oil successfully refilled). The majority of adverse events (AEs) were transient and mostly of mild to moderate intensity. Conclusions: Psychophysical and subjective data show that RETINA IMPLANT Alpha AMS is reliable, well tolerated and can restore limited visual functions in blind patients with degenerations of the outer retina. Compared with the previous implant Alpha IMS, longevity of the new implant Alpha AMS has been considerably improved. Alpha AMS has meanwhile been certified as a commercially available medical device, reimbursed in Germany by the public health system.


The P300 event-related potential in bipolar disorder: A systematic review and meta-analysis.
Neurophysiology including P300, that is a typical index of event-related potential, may be potential biomarkers for bipolar disorder (BD) and it can be useful towards elucidating the pathophysiology of BD. However, previous findings from P300 studies were inconsistent due to the heterogeneity of research methods, which make it difficult to understand the neurobiological significance of them. The aim of this study is to conduct a meta-analysis on P300 in patients with BD. A literature search was conducted using PubMed to identify studies that compared P300 event-related potential between patients with BD and healthy controls (HCs). We analyzed P300 indices such as amplitude and latency of P3a and P3b in auditory or visual paradigms. Further, moderator analyses were conducted to investigate the influence of patient characteristics (i.e. history of psychosis, diagnostic subcategories [BD-I/BD-II], and phase of illness [euthymic, manic, or depressive]) on P300 indices. Out of 124 initial records, we included 30 articles (BD: N = 1331; HCs: N = 1818). Patients with BD showed reduced P3a and P3b amplitude in both paradigms and delayed P3b latency in auditory paradigms compared to HCs. There was no influence on the history of psychosis, diagnostic subcategories, or phase of illness on P300 indices. The difference in medication use was difficult to control and it may affect the results. This meta-analysis provides evidence for P300 abnormalities in patients with BD compared to HCs. Our results suggest that P300 may be trait markers rather than state markers in this illness.


Efficient and high fidelity incorporation of dCTP opposite 7,8-dihydro-8-oxodeoxyguanosine by Sulfolobus solfataricus DNA polymerase Dpo4.
DNA polymerases insert dATP opposite the oxidative damage product 7,8-dihydro-8-oxodeoxyguanosine (8-oxoG) instead of dCTP, to the extent of >90% with some polymerases. Steady-state kinetics with the Y-family Sulfolobus solfataricus DNA polymerase IV (Dpo4) showed 90-fold higher incorporation efficiency of dCTP > dATP opposite 8-oxoG and 4-fold higher efficiency of extension beyond an 8-oxoG:C pair than an 8-oxoG:A pair. The catalytic efficiency for these events (with dCTP or C) was similar for G and 8-oxoG templates. Mass spectral analysis of extended DNA primers showed >/=95% incorporation of dCTP > dATP opposite 8-oxoG. Pre-steady-state kinetics showed faster rates of dCTP incorporation opposite 8-oxoG than G. The measured K(d)(,dCTP) was 15-fold lower for an oligonucleotide containing 8-oxoG than with G. Extension beyond an 8-oxoG:C pair was similar to G:C and faster than for an 8-oxoG:A pair, in contrast to other polymerases. The E(a) for dCTP insertion opposite 8-oxoG was lower than for opposite G. Crystal structures of Dpo4 complexes with oligonucleotides were solved with C, A, and G nucleoside triphosphates placed opposite 8-oxoG. With ddCTP, dCTP, and dATP the phosphodiester bonds were formed even in the presence of Ca(2+). The 8-oxoG:C pair showed classic Watson-Crick geometry; the 8-oxoG:A pair was in the syn:anti configuration, with the A hybridized in a Hoogsteen pair with 8-oxoG. With dGTP placed opposite 8-oxoG, pairing was not to the 8-oxoG but to the 5' C (and in classic Watson-Crick geometry), consistent with the low frequency of this frameshift event observed in the catalytic assays.


Insights into plant consciousness from neuroscience, physics and mathematics: a role for quasicrystals?
There is considerable debate over whether plants are conscious and this, indeed, is an important question. Here I look at developments in neuroscience, physics and mathematics that may impact on this question. Two major concomitants of consciousness in animals are microtubule function and electrical gamma wave synchrony. Both these factors may also play a role in plant consciousness. I show that plants possess aperiodic quasicrystal structures composed of ribosomes that may enable quantum computing, which has been suggested to lie at the core of animal consciousness. Finally I look at whether a microtubule fractal suggests that electric current plays a part in conventional neurocomputing processes in plants.


Antiparasitic effect of the Psidium guajava L. (guava) and Psidium brownianum MART. EX DC. (araçá-de-veado) extracts.
In the search for new therapeutic agents against neglected diseases, both aqueous and hydroethanolic extracts from Psidium guajava L. and P. brownianum Mart ex DC leaves were investigated regarding their antiparasitic effect and cytotoxic potential. The extracts were tested at three concentrations (250, 500 and 1000 μg/mL) against Trypanosoma cruzi epimastigote forms (Chagas, 1909), Leishmania braziliensis (Vianna, 1911) and L. infantum promastigotes forms (Nicolle, 1908), as well as against fibroblasts. P. guajava showed no activity against T. cruzi forms, while the hydroethanolic (PBHE), aqueous by decoction (PBAED) and aqueous by infusion (PBAEI) P. browninaum extracts were responsible, respectively, for inhibiting 100, 100 and 92.68% of T. cruzi epimastigote growth at the 1000 μg/mL concentration. The P. brownianum hydroethanolic extract (PBHE) at the highest concentration caused 58.46% death in L. braziliensis, thus demonstrating moderate activity, however when tested against L. infantum, the PBHE inhibited their growth by 37.16%, revealing its low activity. As for the cytotoxicity assays, the P. brownianum aqueous extract by decoction (PBAED) obtained the highest death percentage when compared to the others, causing 90.85% fibroblast mortality at the 1000 μg/mL concentration.


Lumefantrine and Desbutyl-Lumefantrine Population Pharmacokinetic-Pharmacodynamic Relationships in Pregnant Women with Uncomplicated Plasmodium falciparum Malaria on the Thailand-Myanmar Border.
Artemether-lumefantrine is the most widely used antimalarial artemisinin-based combination treatment. Recent studies have suggested that day 7 plasma concentrations of the potent metabolite desbutyl-lumefantrine correlate better with treatment outcomes than those of lumefantrine. Low cure rates have been reported in pregnant women with uncomplicated falciparum malaria treated with artemether-lumefantrine in northwest Thailand. A simultaneous pharmacokinetic drug-metabolite model was developed based on dense venous and sparse capillary lumefantrine and desbutyl-lumefantrine plasma samples from 116 pregnant patients on the Thailand-Myanmar border. The best model was used to evaluate therapeutic outcomes with a time-to-event approach. Lumefantrine and desbutyl-lumefantrine concentrations, implemented in an Emax model, both predicted treatment outcomes, but lumefantrine provided better predictive power. A combined model including both lumefantrine and desbutyl-lumefantrine did not improve the model further. Simulations suggested that cure rates in pregnant women with falciparum malaria could be increased by prolonging the treatment course. (These trials were registered at controlled-trials.com [ISRCTN 86353884].).


Peripheral intravenous catheters in situ for more than 96 h in adults: What factors affect removal?
Peripheral intravenous catheters (PIVC) are widely used in clinical nursing, but indwelling time remains a subject of debate. This study aimed to assess the risk factors for PIVC phlebitis in adults and provide a basis for indwelling time decisions. A total of 189 first-time PIVC patients in the emergency ward were assessed between May and October 2015. Data were retrieved for patient characteristics and PIVC assessment records. This study showed that over two-thirds (67.72%) of PIVCs were removed because of phlebitis, including oedema (37.57%), rubefaction (33.33%), pain (32.28%), slow infusion speed (13.23%) and accidental extrusion (2.12%). PIVC indwelling time in the planned removal group was higher than that obtained for the unplanned removal group: 152.42 (74.58) vs. 94.64 (50.15) h, P < 0.001. At indwelling times > 96 h, 28.57% (n = 54) of catheters caused phlebitis, although 23.28% (n = 44) showed no phlebitis. PIVC phlebitis was associated with treatment with compound amino acid infusion (OR: 2.624), site at the elbow joint (OR: 3.049), haemoglobin level (OR: 2.492), white cell count (OR: 2.196) and catheter size (OR: 1.837). Study findings suggest that PIVC might be used for longer durations based on nursing assessments and health education.


A Versatile and Robust Serine Protease Inhibitor Scaffold from Actinia tenebrosa.
Serine proteases play pivotal roles in normal physiology and a spectrum of patho-physiological processes. Accordingly, there is considerable interest in the discovery and design of potent serine protease inhibitors for therapeutic applications. This led to concerted efforts to discover versatile and robust molecular scaffolds for inhibitor design. This investigation is a bioprospecting study that aims to isolate and identify protease inhibitors from the cnidarian Actinia tenebrosa. The study isolated two Kunitz-type protease inhibitors with very similar sequences but quite divergent inhibitory potencies when assayed against bovine trypsin, chymostrypsin, and a selection of human sequence-related peptidases. Homology modeling and molecular dynamics simulations of these inhibitors in complex with their targets were carried out and, collectively, these methodologies enabled the definition of a versatile scaffold for inhibitor design. Thermal denaturation studies showed that the inhibitors were remarkably robust. To gain a fine-grained map of the residues responsible for this stability, we conducted in silico alanine scanning and quantified individual residue contributions to the inhibitor's stability. Sequences of these inhibitors were then used to search for Kunitz homologs in an A. tenebrosa transcriptome library, resulting in the discovery of a further 14 related sequences. Consensus analysis of these variants identified a rich molecular diversity of Kunitz domains and expanded the palette of potential residue substitutions for rational inhibitor design using this domain.


Curve evolution implementation of the Mumford-Shah functional for image segmentation, denoising, interpolation, and magnification.
In this work, we first address the problem of simultaneous image segmentation and smoothing by approaching the Mumford-Shah paradigm from a curve evolution perspective. In particular, we let a set of deformable contours define the boundaries between regions in an image where we model the data via piecewise smooth functions and employ a gradient flow to evolve these contours. Each gradient step involves solving an optimal estimation problem for the data within each region, connecting curve evolution and the Mumford-Shah functional with the theory of boundary-value stochastic processes. The resulting active contour model offers a tractable implementation of the original Mumford-Shah model (i.e., without resorting to elliptic approximations which have traditionally been favored for greater ease in implementation) to simultaneously segment and smoothly reconstruct the data within a given image in a coupled manner. Various implementations of this algorithm are introduced to increase its speed of convergence. We also outline a hierarchical implementation of this algorithm to handle important image features such as triple points and other multiple junctions. Next, by generalizing the data fidelity term of the original Mumford-Shah functional to incorporate a spatially varying penalty, we extend our method to problems in which data quality varies across the image and to images in which sets of pixel measurements are missing. This more general model leads us to a novel PDE-based approach for simultaneous image magnification, segmentation, and smoothing, thereby extending the traditional applications of the Mumford-Shah functional which only considers simultaneous segmentation and smoothing.


Frequency of monoclonal B-cell lymphocytosis in relatives of patients with chronic lymphocytic leukemia.
Monoclonal B-cell lymphocytosis is a symptom free condition characterized by the circulation of small clonal population of B lymphocytes in peripheral blood (less than 5x10(9)/L) expressing an immunophenotype similar to chronic lymphocytic leukemia. Different studies based on big hospital series have manifested a higher risk in subjects with monoclonal B-cell lymphocytosis to progress to a chronic lymphocytic leukemia. The behavior of this hematologic entity is unknown therefore its frequency in sporadic chronic lymphocytic leukemia patient relatives was determined. Transversal descriptive study, 8 color flow cytometry was performed using two of the tubes of the Euro Flow recommended panel, with modifications, for the diagnose of chronic lymphoproliferative disorders of B lymphocytes; besides, a fluorescence in situ hybridization was performed. univariate and bivariate analyses of the information were performed. Monoclonal B-cell lymphocytosis frequency found in 51 analyzed relatives was 2%, it was a female participant, 59 years old, with a total leukocyte count of 7.7x10(9)/L and a B lymphocyte count of 0.124x10(9)/L; from these, 0.04x10(9)/L were clonal cells with restrictions of the kappa light chain. Rearrangements of the IGH gene (14q32) were found. Monoclonal B-cell lymphocytosis was detected in one relative of a patient with sporadic chronic lymphocytic leukemia in a frequency similar to the one reported in general population.


The acute effects of a caffeine-containing supplement on strength, muscular endurance, and anaerobic capabilities.
The purpose of this study was to examine the acute effects of a caffeine-containing supplement on upper- and lower-body strength and muscular endurance as well as anaerobic capabilities. Thirty-seven resistance-trained men (mean +/- SD, age: 21 +/- 2 years) volunteered to participate in this study. On the first laboratory visit, the subjects performed 2 Wingate Anaerobic Tests (WAnTs) to determine peak power (PP) and mean power (MP), as well as tests for 1 repetition maximum (1RM), dynamic constant external resistance strength, and muscular endurance (TOTV; total volume of weight lifted during an endurance test with 80% of the 1RM) on the bilateral leg extension (LE) and free-weight bench press (BP) exercises. Following a minimum of 48 hours of rest, the subjects returned to the laboratory for the second testing session and were randomly assigned to 1 of 2 groups: a supplement group (SUPP; n = 17), which ingested a caffeine-containing supplement, or a placebo group (PLAC; n = 20), which ingested a cellulose placebo. One hour after ingesting either the caffeine-containing supplement or the placebo, the subjects performed 2 WAnTs and were tested for 1RM strength and muscular endurance on the LE and BP exercises. The results indicated that there was a significant (p < 0.05) increase in BP 1RM for the SUPP group, but not for the PLAC group. The caffeine-containing supplement had no effect, however, on LE 1RM, LE TOTV, BP TOTV, PP, and MP. Thus, the caffeine-containing supplement may be an effective supplement for increasing upper-body strength and, therefore, could be useful for competitive and recreational athletes who perform resistance training.


Protective action by methylprednisolone, allopurinol and indomethacin against stroke-induced damage to adenylate cyclase in gerbil cerebral cortex.
Adenylate cyclase activity was investigated in either homogenate or particulate fractions from the frontal cerebral cortex of the gerbil following five experimental conditions of bilateral ischemia. After periods of 15 min ischemia, 15 min ischemia plus 15 min of recirculation or 60 min ischemia the enzyme generally displayed enhanced responses to GTP, norepinephrine (NE), dopamine (DA), NE + GTP and DA + GTP. Pretreatment of the gerbils with methylprednisolone, allopurinol or indomethacin did not significantly influence the outcome of these findings. When the animals were subjected to 60 min ischemia plus 15 min of reflow, enzyme responses to the stimulatory agents including forskolin and NaF were all reduced. Pretreatment with methylprednisolone, allopurinol or indomethacin prevented the damage to adenylate cyclase in the 60 min ischemia plus 15 min reflow animals. When animals were made ischemic for 15 min followed by one week of recovery, enzyme sensitivity to GTP, calmodulin-Ca++, NE, combinations thereof and forskolin were reduced in only the particulate fractions. Enzyme damage was reversed following methylprednisolone. Enzyme damage may result from generation of free radicals during reflow and drugs that either inhibit synthesis pathways generating free radicals, stabilize cell membranes or act as free radical scavengers may be therapeutically beneficial under specific conditions of stroke.


Dietary acrylamide and the risk of pancreatic cancer in the International Pancreatic Cancer Case-Control Consortium (PanC4).
Occupational exposure to acrylamide was associated with excess mortality from pancreatic cancer, though in the absence of dose-risk relationship. Few epidemiological studies have examined the association between acrylamide from diet and pancreatic cancer risk. We considered this issue in a combined set of 1975 cases of pancreatic cancer and 4239 controls enrolled in six studies of the Pancreatic Cancer Case-Control Consortium (PanC4). We calculated pooled odds ratios (ORs) and their 95% confidence intervals (CI) by estimating study-specific ORs through multivariate unconditional logistic regression models and pooling the obtained estimates using random-effects models. Compared with the lowest level of estimated dietary acrylamide intake, the pooled ORs were 0.97 (95% CI, 0.79-1.19) for the second, 0.91 (95% CI, 0.71-1.16) for the third, and 0.92 (95% CI, 0.66-1.28) for the fourth (highest) quartile of intake. For an increase of 10 µg/day of acrylamide intake, the pooled OR was 0.96 (95% CI, 0.87-1.06), with heterogeneity between estimates (I2 = 67%). Results were similar across various subgroups, and were confirmed when using a one-stage modelling approach. This PanC4 pooled-analysis found no association between dietary acrylamide and pancreatic cancer.


Triticum mosaic virus Isolates in the Southern Great Plains.
In 2006, a previously unknown wheat (Triticum aestivum) virus was discovered in Western Kansas and given the name Triticum mosaic virus (TriMV). TriMV has since been found in wheat samples isolated all across the Great Plains. Even though it can infect singularly, TriMV is mostly found with Wheat streak mosaic virus (WSMV) as a co-infection. The potential for TriMV to cause economic loss is significant, but very little is known about the virus. The objective of this study was to survey the TriMV population for genetic variation by nucleotide sequencing of isolates across a geographical region. A secondary objective was to characterize the WSMV isolates that are being co-transmitted with TriMV. Fourteen different TriMV isolations were taken from locations in Texas, Oklahoma, and Kansas, and the coat protein cDNA was sequenced. Thirteen nucleotide differences were found in the TriMV isolates, of which three induce amino acid changes. WSMV isolates had 65 nucleotide changes when compared to WSMV Sydney81. Our results indicate the TriMV virus population has minimal amounts of sequence variation and no singular WSMV genotype is specifically associated with TriMV co-infection. Based on the isolates analyzed, it appears that the field population of TriMV is very homogeneous.


A multi-metric approach to investigate the effects of weather conditions on the demographic of a terrestrial mammal, the european badger (Meles meles).
Models capturing the full effects of weather conditions on animal populations are scarce. Here we decompose yearly temperature and rainfall into mean trends, yearly amplitude of change and residual variation, using daily records. We establish from multi-model inference procedures, based on 1125 life histories (from 1987 to 2008), that European badger (Meles meles) annual mortality and recruitment rates respond to changes in mean trends and to variability in proximate weather components. Variation in mean rainfall was by far the most influential predictor in our analysis. Juvenile survival and recruitment rates were highest at intermediate levels of mean rainfall, whereas low adult survival rates were associated with only the driest, and not the wettest, years. Both juvenile and adult survival rates also exhibited a range of tolerance for residual standard deviation around daily predicted temperature values, beyond which survival rates declined. Life-history parameters, annual routines and adaptive behavioural responses, which define the badgers' climatic niche, thus appear to be predicated upon a bounded range of climatic conditions, which support optimal survival and recruitment dynamics. That variability in weather conditions is influential, in combination with mean climatic trends, on the vital rates of a generalist, wide ranging and K-selected medium-sized carnivore, has major implications for evolutionary ecology and conservation.


Psychological therapies for people with intellectual disabilities: a systematic review and meta-analysis.
The aim of this study was to evaluate the efficacy of psychological therapies for people with intellectual disabilities (IDs) through a systematic review and meta-analysis of the current literature. A comprehensive literature search identified 143 intervention studies. Twenty-two trials were eligible for review, and 14 of these were subsequently included in the meta-analysis. Many studies did not include adequate information about their participants, especially the nature of their IDs; information about masked assessment, and therapy fidelity was also lacking. The meta-analysis yielded an overall moderate between-group effect size, g=.682, while group-based interventions had a moderate but smaller treatment effect than individual-based interventions. Cognitive-behaviour therapy (CBT) was efficacious for both anger and depression, while interventions aimed at improving interpersonal functioning were not effectual. When CBT was excluded, there was insufficient evidence regarding the efficacy of other psychological therapies, or psychological therapies intended to treat mental health problems in children and young people with IDs. Adults with IDs and concurrent mental health problems appear to benefit from psychological therapies. However, clinical trials need to make use of improved reporting standards and larger samples.


Peroxynitrite-induced oligodendrocyte toxicity is not dependent on poly(ADP-ribose) polymerase activation.
Oligodendrocyte loss is a characteristic feature of several CNS disorders, including multiple sclerosis (MS) and spinal cord injury. However, the mechanisms responsible for oligodendrocyte destruction remain undefined. As recent studies have implicated peroxynitrite in the pathogenesis of both spinal cord injury and MS, we have examined whether peroxynitrite may mediate at least some of the oligodendrocyte damage and demyelination observed in these conditions. Primary rat oligodendrocytes were exposed to authentic peroxynitrite in vitro and assessed for cytotoxicity. Mitochondrial function, measured by the reduction of MTT to formazan, and mitochondrial membrane potential were used as indicators of cell viability. Cell death was quantitated by measuring either the release of lactate dehydrogenase from, or the uptake of propidium iodide into, damaged and dying cells. Peroxynitrite dose-dependently reduced the viability of primary oligodendrocytes and induced cell death. Furthermore, peroxynitrite significantly increased DNA strand breakage and the activity of poly(ADP-ribose) polymerase (PARP) in oligodendrocyte cultures. To identify whether PARP activation plays a role in peroxynitrite-induced oligodendrocyte toxicity, we examined the effects of the PARP inhibitors 3-aminobenzamide (3AB) and 5-iodo-6-amino-1,2-benzopyrone (INH(2)BP) on mitochondrial function and cell death in oligodendrocytes. The presence of 3AB and INH(2)BP did not protect oligodendrocytes from peroxynitrite-induced cytotoxicity. However, both compounds significantly reduced PARP activity in these cells. Primary oligodendrocytes generated from PARP-deficient mice were also highly susceptible to peroxynitrite-induced cell death. Therefore, our results show that peroxynitrite exerts cytotoxic effects on oligodendrocytes in vitro independently of PARP activation.


Activation of transcription in cell-free extracts by a novel cAMP-responsive sequence from the bovine CYP17 gene.
An in vitro transcription system has been developed to study the transcriptional control of steroid hydroxylase gene expression in steroidogenic tissues using upstream sequences of these genes in rabbit beta-globin promoter/reporter constructs. Analysis of deletion mutants of the upstream region of the bovine CYP17 (p45017 alpha) gene in whole cell extracts prepared from mouse Y1 adrenocortical cells revealed that sequences activating transcription in vitro are identical to those previously identified in vivo, namely the novel cAMP-responsive sequences crsI (-243 to -225 base pairs) and crsII (-80 to -40 base pairs). These sequences do not contain the known cAMP response elements, CRE or AP-2. In similar extracts from nonsteroidogenic mouse SVT2 cells, crsI was unable to activate transcription. Partially purified protein factor(s) binding to crsI were prepared from Y1 extracts by crsI-specific DNA affinity chromatography. This fraction activated transcription in vitro in SVT2 extracts only when crsI was present in the template. This in vitro transcription system will permit further investigation of crsI and its cognate factors and of the transcriptional regulation of the other steroid hydroxylase genes.


Automatic segmentation and classification of outdoor images using neural networks.
The paper describes how neural networks may be used to segment and label objects in images. A self-organising feature map is used for the segmentation phase, and we quantify the quality of the segmentations produced as well as the contribution made by colour and texture features. A multi-layer perception is trained to label the regions produced by the segmentation process. It is shown that 91.1% of the image area is correctly classified into one of eleven categories which include cars, houses, fences, roads, vegetation and sky.


Effect of Liquid Swine Manure on Hatch and Viability of Heterodera glycines.
Experiments were conducted in the laboratory and greenhouse to determine the effect of raw and anaerobically digested liquid swine manures on the hatch and viability of Heterodera glycines, the soybean cyst nematode. Anaerobic digestion was performed for 15 and 35 days to enrich volatile fatty acids (VFA) and ammonium (NH(4) (+)), respectively. All filtrates of the raw, VFA-enriched, and NH(4) (+)-enriched manures at 10(-1) to 250(-1) dilutions inhibited H. glycines hatch, and the reduction of hatch was increased with increasing concentration of the manure. Cumulative hatch at day 21 was only 2.1% to 3.7% in the 10(-1) dilution manures, while the hatch in water was 21% to 27.3%. The high concentrations appeared to be lethal to some eggs. Most second-stage juveniles (J2) of H. glycines were killed when incubated for 8 hours in the manure filtrate at the original concentration (>90% mortality) or for 48 hours at the 64(-1) dilution (> 82% mortality). When J2 were treated with the manures at 10(-1) to 250(-1) dilutions for 4 hours, only the 10(-1) dilution of VFA-enriched and raw manures resulted in a lower number of J2 that penetrated soybean roots as compared with lower concentrations. The VFA-enriched manure was the best, raw manure intermediate, and NH(4) (+)-enriched manure the least effective in inhibiting H. glycines hatch and killing eggs and J2.


Critical evaluation of the composition and use of topical fluorides.
This report reviews the current status of various topical fluoride systems commonly used for caries prevention; these systems include professional fluoride applications, prophylaxis pastes, varnishes, rinses, gels, and dentifrices. Although the cariostatic benefits of professional applications are well-established from early studies, the magnitude of benefit relative to other preventive measures needs to be re-assessed under current conditions. Significant cariostatic benefits from fluoride prophylaxis pastes remain to be established, as does the relative efficacy of fluoride varnishes in comparison with other fluoride measures. The established cariostatic activity of fluoride rinses with supervised usage needs confirmation under normal home usage conditions. There is inadequate evidence to justify either increasing or decreasing the concentration of fluoride in conventional dentifrices whose efficacy is well-established. Based on limited clinical data and supportive laboratory data, fluoride appears to be effective for preventing root-surface caries.


Indications for and outcomes of limb amputation in goats and sheep.
OBJECTIVE To characterize indications for and clinical outcomes of limb amputation in goats and sheep. DESIGN Retrospective case series and observational study. ANIMALS Goats (n = 15) and sheep (7) that underwent partial or complete limb amputation at the University of California-Davis Veterinary Medical Teaching Hospital from January 1, 1985, through December 31, 2015. PROCEDURES Medical records of qualifying goats and sheep were reviewed and data extracted regarding signalment, use of animal, characteristics of the amputated limb, duration of hospitalization, outcome, and surgery-associated complications. The Fisher exact test was performed to compare postsurgical complication rates between various groups. RESULTS Reasons for limb amputation included trauma (ie, fracture, dog bite, and wire-fence injury) and infectious or degenerative disease (ie, osteomyelitis and osteoarthritis). Median (range) hospitalization period for goats and sheep was 7.5 days (1 to 63 days) and 8 days (3 to 20 days), respectively. Most (7/9) of the postsurgical complications were noticed > 2 months after amputation and included uncoordinated gait, tendon breakdown and laxity of the contralateral limb, chronic lameness, surgical site infection, chronic intermittent pain, and angular limb deformity. No association was identified between postsurgical complication rate in goats and body weight category (< 20 vs ≥ 20 kg [44 lb]), age category (< 2 vs ≥ 2 years), and amputated limb type (forelimb vs hind limb). CONCLUSIONS AND CLINICAL RELEVANCE Clinical outcomes for goats and sheep following limb amputation varied and were seemingly case dependent. Amputation should be considered when preservation of the whole limb is not possible. Veterinarians should advise owners that limb amputation may result in various complications and recommend aftercare treatment such as physiotherapy.


Neurological examination of the upper limb: a study of construct validity.
We have previously demonstrated that neurological individual findings and patterns can be reliably assessed in the examination of the upper limb and also that they are related to pain, weakness, and/or numbness/tingling. This study aimed to study further aspects of the construct validity of the neurological examination. Blinded to patient-characteristics, two examiners assessed the function of 16 muscles, the sensibility in 7 territories, and the nerve-mechanosensitivity at 20 locations in 82 upper limbs. Based on anatomical patterns and pre-designed algorithms, one or both examiners rated neuropathy as "possible" or "definite" in 40 limbs and also determined the location( s). We developed and tested hypotheses on anatomically and regionally related locations of nerve afflictions (a selective vulnerability of neurons, double and multiple crush, and a tendency to regional spread) and examined the stability of the internal structure of the constructs in different situations. The interrelations of findings were analyzed by hypothesis testing and factor analyses, and the homogeneity of location profiles was analyzed by a conditional likelihood test. Out of 30 limbs with related locations of neuropathy, the findings of each examiner correlated positively (gamma > 0.35) in 22/25, respectively. The patterns of the interrelations identified by the two examiners were similar, with no evidence of any heterogeneity of location profiles for either examiner. This study supports the validity of the physical examination. However, feasibility of its application requires the demonstration of further aspects of construct validity and a favorable influence on patient-management and/or prevention.


Anesthetic induction with guaifenesin and propofol in adult horses.
To evaluate whether guaifenesin can prevent adverse anesthetic induction events caused by propofol and whether a guaifenesin-propofol induction combination has brief cardiovascular effects commensurate with rapid drug washout. 8 healthy adult horses. Guaifenesin was administered IV for 3 minutes followed by IV injection of a bolus of propofol (2 mg/kg). Additional propofol was administered if purposeful movement was detected. Anesthesia was maintained for 2 hours with isoflurane or sevoflurane at 1.2 times the minimum alveolar concentration with controlled normocapnic ventilation. Normotension was maintained via a dobutamine infusion. Plasma concentrations of propofol and guaifenesin were measured every 30 minutes. Mean ± SD guaifenesin and propofol doses inducing anesthesia in half of the horses were 73 ± 18 mg/kg and 2.2 ± 0.3 mg/kg, respectively. No adverse anesthetic induction events were observed. By 70 minutes, there was no significant temporal change in the dobutamine infusion rate required to maintain normotension for horses anesthetized with isoflurane or sevoflurane. Mean plasma guaifenesin concentrations were 122 ± 30 μM, 101 ± 33 μM, 93 ± 28 μM, and 80 ± 24 μM at 30, 60, 90, and 120 minutes after anesthetic induction, respectively. All plasma propofol concentrations were below the limit of quantitation. Guaifenesin prevented adverse anesthetic induction events caused by propofol. Guaifenesin (90 mg/kg) followed by propofol (3 mg/kg) should be sufficient to immobilize > 99% of calm healthy adult horses. Anesthetic drug washout was rapid, and there was no change in inotrope requirements after anesthesia for 70 minutes.


A Role for the Brain RAS in Alzheimer's and Parkinson's Diseases.
The brain renin-angiotensin system (RAS) has available the necessary functional components to produce the active ligands angiotensins II (AngII), angiotensin III, angiotensins (IV), angiotensin (1-7), and angiotensin (3-7). These ligands interact with several receptor proteins including AT1, AT2, AT4, and Mas distributed within the central and peripheral nervous systems as well as local RASs in several organs. This review first describes the enzymatic pathways in place to synthesize these ligands and the binding characteristics of these angiotensin receptor subtypes. We next discuss current hypotheses to explain the disorders of Alzheimer's disease (AD) and Parkinson's disease (PD), as well as research efforts focused on the use of angiotensin converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs), in their treatment. ACE inhibitors and ARBs are showing promise in the treatment of several neurodegenerative pathologies; however, there is a need for the development of analogs capable of penetrating the blood-brain barrier and acting as agonists or antagonists at these receptor sites. AngII and AngIV have been shown to play opposing roles regarding memory acquisition and consolidation in animal models. We discuss the development of efficacious AngIV analogs in the treatment of animal models of AD and PD. These AngIV analogs act via the AT4 receptor subtype which may coincide with the hepatocyte growth factor/c-Met receptor system. Finally, future research directions are described concerning new approaches to the treatment of these two neurological diseases.


Investigation of sex-specific effects of apolipoprotein E on severity of EAE and MS.
Despite pleiotropic immunomodulatory effects of apolipoprotein E (apoE) in vitro, its effects on the clinical course of experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS) are still controversial. As sex hormones modify immunomodulatory apoE functions, they may explain contentious findings. This study aimed to investigate sex-specific effects of apoE on disease course of EAE and MS. MOG(35-55) induced EAE in female and male apoE-deficient mice was assessed clinically and histopathologically. apoE expression was investigated by qPCR. The association of the MS severity score (MSSS) and APOE rs429358 and rs7412 was assessed across 3237 MS patients using linear regression analyses. EAE disease course was slightly attenuated in male apoE-deficient (apoE (-/-) ) mice compared to wildtype mice (cumulative median score: apoE (-/-) = 2 [IQR 0.0-4.5]; wildtype = 4 [IQR 1.0-5.0]; n = 10 each group, p = 0.0002). In contrast, EAE was more severe in female apoE (-/-) mice compared to wildtype mice (cumulative median score: apoE (-/-) = 3 [IQR 2.0-4.5]; wildtype = 3 [IQR 0.0-4.0]; n = 10, p = 0.003). In wildtype animals, apoE expression during the chronic EAE phase was increased in both females and males (in comparison to naïve animals; p < 0.001). However, in MS, we did not observe a significant association between MSSS and rs429358 or rs7412, neither in the overall analyses nor upon stratification for sex. apoE exerts moderate sex-specific effects on EAE severity. However, the results in the apoE knock-out model are not comparable to effects of polymorphic variants in the human APOE gene, thus pinpointing the challenge of translating findings from the EAE model to the human disease.


Analyzing clinical trial outcomes based on incomplete daily diary reports.
A case study is presented assessing the impact of missing data on the analysis of daily diary data from a study evaluating the effect of a drug for the treatment of insomnia. The primary analysis averaged daily diary values for each patient into a weekly variable. Following the commonly used approach, missing daily values within a week were ignored provided there was a minimum number of diary reports (i.e., at least 4). A longitudinal model was then fit with treatment, time, and patient-specific effects. A treatment effect at a pre-specified landmark time was obtained from the model. Weekly values following dropout were regarded as missing, but intermittent daily missing values were obscured. Graphical summaries and tables are presented to characterize the complex missing data patterns. We use multiple imputation for daily diary data to create completed data sets so that exactly 7 daily diary values contribute to each weekly patient average. Standard analysis methods are then applied for landmark analysis of the completed data sets, and the resulting estimates are combined using the standard multiple imputation approach. The observed data are subject to digit heaping and patterned responses (e.g., identical values for several consecutive days), which makes accurate modeling of the response data difficult. Sensitivity analyses under different modeling assumptions for the data were performed, along with pattern mixture models assessing the sensitivity to the missing at random assumption. The emphasis is on graphical displays and computational methods that can be implemented with general-purpose software. Copyright © 2016 John Wiley & Sons, Ltd.


Oral anticoagulant therapy: Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines.
The objective of this article is to summarize the published literature concerning the pharmacokinetics and pharmacodynamics of oral anticoagulant drugs that are currently available for clinical use and other aspects related to their management. We carried out a standard review of published articles focusing on the laboratory and clinical characteristics of the vitamin K antagonists; the direct thrombin inhibitor, dabigatran etexilate; and the direct factor Xa inhibitor, rivaroxaban The antithrombotic effect of each oral anticoagulant drug, the interactions, and the monitoring of anticoagulation intensity are described in detail and discussed without providing specific recommendations. Moreover, we describe and discuss the clinical applications and optimal dosages of oral anticoagulant therapies, practical issues related to their initiation and monitoring, adverse events such as bleeding and other potential side effects, and available strategies for reversal. There is a large amount of evidence on laboratory and clinical characteristics of vitamin K antagonists. A growing body of evidence is becoming available on the first new oral anticoagulant drugs available for clinical use, dabigatran and rivaroxaban.


Food for thought ... considerations and guidelines for basic test method descriptions in toxicology.
The development and application of new test systems and test methods is central to the progress of in vitro toxicology. In order to live up to the future challenges, it is important to use the vast knowledge of adjoining fields, such as cell biology or developmental biology, and to attract specialists from such areas to develop new methods. Not all of them bring with them the necessary experience and training required for the development of toxicological test systems. Thus, promising new biological test systems sometimes still require additional considerations to become successful. Sometimes even the referees of scientific journals and their editors seem to lack judgement guidelines for minimum performance standards. Here we provide a list of points to be considered for the establishment of a test method. The chapters range from the explanation of the meaning of positive controls, performance standards or signal-noise ratios to a discussion of statistical considerations, suitable solvents and data display formats. The considerations are simple and expressed in a non-mathematical format, with a strong focus on plausibility and common sense. The major intention of this article is to provide a compilation of important issues requiring consideration. Whether they apply to a specific system and whether action is required must be determined by individual judgement.


Differentiation of Mycobacterium tuberculosis complex by PCR amplification of genomic regions of difference.
Differentiation of members of the Mycobacterium tuberculosis complex by conventional mycobacteriological methods is time consuming, making surveillance of species-specific disease difficult. A two-step, multiplex polymerase chain reaction (PCR) method based on genomic regions of difference (RD1, RD1(mic), RD2(seal), RD4, RD9 and RD12) was developed for the differentiation of M. canettii, M. tuberculosis, M. africanum, M. microti, M. pinnipedii, M. caprae, M. bovis and M. bovis BCG. The size of the respective multiplex PCR amplification products corresponded to the presence of the different M. tuberculosis complex members. This method allows for rapid differentiation, making it suitable for routine laboratories and surveillance purposes.


Effect on ejaculatory performance and semen parameters of sexually-satiated male goats (Capra hircus) after changing the stimulus female.
The association between stimulation of sexual activity and semen characteristics of sexually mature goats was evaluated. Nine 2-year-old criollo bucks were used. Each buck was subjected to 8-weekly trials in which one of the following two treatments was alternately applied. In treatment 1, males were individually exposed to the same estrus-induced female during a 4-h period, and ejaculates were collected and analyzed throughout. Treatment 2 was the same, except that a different doe replaced the stimulus animal for the third and fourth hours. The total number of ejaculations and total sperm number for four sets of data was different (P<0.01) for treatments 1 and 2 (184 versus 252 ejaculations with a total of 354.3 versus 477.1 billion sperm, respectively). In treatment 2, when first and last ejaculation of each male with the (original) stimulus animal were compared, total sperm per ejaculate decreased (P=0.08) from 4.14+/-3.8 x 10(9) to 0.77+/-0.7 x 10(9), while this value increased (P=0.05) to 3.04+/-2.3 x 10(9) after a new female was introduced, which represented a recovery of 67.35%. All males achieved a first ejaculation with the original stimulus animal while on average, only three achieved a seventh service. After changing the stimulus animal, all males ejaculated again. It is concluded that changing the stimulus animal after a 2-h continuous exposure to an estrous female stimulates sexual activity and increases sperm output in male goats.