Transcriptomics and methylomics of CD4-positive T cells in arsenic-exposed women.
Arsenic, a carcinogen with immunotoxic effects, is a common contaminant of drinking water and certain food worldwide. We hypothesized that chronic arsenic exposure alters gene expression, potentially by altering DNA methylation of genes encoding central components of the immune system. We therefore analyzed the transcriptomes (by RNA sequencing) and methylomes (by target-enrichment next-generation sequencing) of primary CD4-positive T cells from matched groups of four women each in the Argentinean Andes, with fivefold differences in urinary arsenic concentrations (median concentrations of urinary arsenic in the lower- and high-arsenic groups: 65 and 276 μg/l, respectively). Arsenic exposure was associated with genome-wide alterations of gene expression; principal component analysis indicated that the exposure explained 53% of the variance in gene expression among the top variable genes and 19% of 28,351 genes were differentially expressed (false discovery rate <0.05) between the exposure groups. Key genes regulating the immune system, such as tumor necrosis factor alpha and interferon gamma, as well as genes related to the NF-kappa-beta complex, were significantly downregulated in the high-arsenic group. Arsenic exposure was associated with genome-wide DNA methylation; the high-arsenic group had 3% points higher genome-wide full methylation (>80% methylation) than the lower-arsenic group. Differentially methylated regions that were hyper-methylated in the high-arsenic group showed enrichment for immune-related gene ontologies that constitute the basic functions of CD4-positive T cells, such as isotype switching and lymphocyte activation and differentiation. In conclusion, chronic arsenic exposure from drinking water was related to changes in the transcriptome and methylome of CD4-positive T cells, both genome wide and in specific genes, supporting the hypothesis that arsenic causes immunotoxicity by interfering with gene expression and regulation.


Chronic acid water feeding protects mice against lethal gut-derived sepsis due to Pseudomonas aeruginosa.
Acidified feeding formulas have been proposed as a method of controlling gastrointestinal colonization and nosocomial infection in critically ill patients. We examined possible mechanisms by which chronic acid water feeding might protect the host against lethal gut derived sepsis by assessing its effect on both local intestinal epithelial barrier function to bacteria as well as on local and systemic heat shock protein expression. Heat shock protein expression measured by immunoblot demonstrated that HSP25 was increased in the stomach, aorta and kidney of mice chronically fed acid water (8 weeks) compared to tap water fed controls. HSP72 expression was also increased in the aorta of mice drinking acid water. The protein content of cecum and its barrier function were enhanced in mice ingesting acidified water. The direct effect of an acid environment on intestinal epithelial barrier function was tested in cultured human intestinal epithelial cells. An acidified environment protected against bacterial mediated disruption of the intestinal epithelial barrier. Finally, the protective effect of chronic acid water feeding on gut-derived sepsis due to P. aeruginosa was tested in mice. Chronic acid water feeding protected mice from the lethal gut derived sepsis due to P. aeruginosa.


Increased nitric oxide production in hypotensive hemodialysis patients.
A subset of patients on long-term hemodialysis have sustained hypotension, defined as a predialysis systolic pressure of < 100 mmHg. To determine the role of nitric oxide (NO), an important vasodilator, in this condition, the authors measured the plasma levels of nitrite (NO2-) and nitrate (NO3-), the known NO metabolites taken as an index of NO production, in 10 hypotensive patients on long-term hemodialysis. None of them had diabetes, cirrhosis of the liver, congestive heart failure, or infection. Fifteen age and gender-matched normotensive patients on hemodialysis were selected as control subjects. Measurements of plasma levels of nitrite and nitrate based on the Greiss reaction were made. There was no significant difference in hematocrit, serum intact parathyroid hormone, total calcium, inorganic phosphorus, albumin, heart rate, cardiac index, or interdialysis weight gain between these two groups. Plasma nitrite and nitrate levels did not correlate with either predialysis serum creatinine or blood urea nitrogen. The mean arterial pressure (MAP) was significantly lower and plasma nitrite and nitrate levels were significantly higher in chronic hypotensive patients than in normotensive patients (MAP: 68.30 +/- 3.24 mmHg vs 95.20 +/- 2.44 mmHg, p < 0.001; plasma nitrite and nitrate: 72.49 +/- 14.41 mumol/L vs 36.42 +/- 5.45 mumol/L, p < 0.05). In addition, MAP from hypotensive and normotensive patients on hemodialysis was inversely correlated with plasma levels of nitrite and nitrate (r = -0.54, p < 0.01). It was concluded that enhanced NO production in this subset of patients on hemodialysis may contribute to their chronic hypotension.


High-Throughput Sequencing Analysis of the Actinobacterial Spatial Diversity in Moonmilk Deposits.
Moonmilk are cave carbonate deposits that host a rich microbiome, including antibiotic-producing Actinobacteria, making these speleothems appealing for bioprospecting. Here, we investigated the taxonomic profile of the actinobacterial community of three moonmilk deposits of the cave "Grotte des Collemboles" via high-throughput sequencing of 16S rRNA amplicons. Actinobacteria was the most common phylum after Proteobacteria, ranging from 9% to 23% of the total bacterial population. Next to actinobacterial operational taxonomic units (OTUs) attributed to uncultured organisms at the genus level (~44%), we identified 47 actinobacterial genera with Rhodoccocus (4 OTUs, 17%) and Pseudonocardia (9 OTUs, ~16%) as the most abundant in terms of the absolute number of sequences. Streptomycetes presented the highest diversity (19 OTUs, 3%), with most of the OTUs unlinked to the culturable Streptomyces strains that were previously isolated from the same deposits. Furthermore, 43% of the OTUs were shared between the three studied collection points, while 34% were exclusive to one deposit, indicating that distinct speleothems host their own population, despite their nearby localization. This important spatial diversity suggests that prospecting within different moonmilk deposits should result in the isolation of unique and novel Actinobacteria. These speleothems also host a wide range of non-streptomycetes antibiotic-producing genera, and should therefore be subjected to methodologies for isolating rare Actinobacteria.


Hypothalamic AMP-activated protein kinase regulates glucose production.
The fuel sensor AMP-activated protein kinase (AMPK) in the hypothalamus regulates energy homeostasis by sensing nutritional and hormonal signals. However, the role of hypothalamic AMPK in glucose production regulation remains to be elucidated. We hypothesize that bidirectional changes in hypothalamic AMPK activity alter glucose production. To introduce bidirectional changes in hypothalamic AMPK activity in vivo, we first knocked down hypothalamic AMPK activity in male Sprague-Dawley rats by either injecting an adenovirus expressing the dominant-negative form of AMPK (Ad-DN AMPKα2 [D(157)A]) or infusing AMPK inhibitor compound C directly into the mediobasal hypothalamus. Next, we independently activated hypothalamic AMPK by delivering either an adenovirus expressing the constitutive active form of AMPK (Ad-CA AMPKα1(312) [T172D]) or the AMPK activator AICAR. The pancreatic (basal insulin)-euglycemic clamp technique in combination with the tracer-dilution methodology was used to assess the impact of alternations in hypothalamic AMPK activity on changes in glucose kinetics in vivo. Injection of Ad-DN AMPK into the hypothalamus knocked down hypothalamic AMPK activity and led to a significant suppression of glucose production with no changes in peripheral glucose uptake during the clamps. In parallel, hypothalamic infusion of AMPK inhibitor compound C lowered glucose production as well. Conversely, molecular and pharmacological activation of hypothalamic AMPK negated the ability of hypothalamic nutrients to lower glucose production. These data indicate that changes in hypothalamic AMPK activity are sufficient and necessary for hypothalamic nutrient-sensing mechanisms to alter glucose production in vivo.


Circadian variation in fibrinolytic activity in patients with variant angina.
Coronary artery spasm induces activation of the coagulation system. Turnover and maintenance of thrombus depend not only on formation but also on lysis. The relation between coronary spasm and fibrinolytic system has not been elucidated. To examine whether there is impairment of or a circadian variation in fibrinolytic activity in patients with variant angina. Plasminogen activator inhibitor (PAI) activity and concentrations of tissue plasminogen activator (t-PA) antigen were measured in venous plasma samples taken at 2200, 0600, and 1400 and 24 h Holter tapes were recorded in 15 patients with variant angina, 12 patients with stable exertional angina, and 12 controls. There were significant circadian variations in PAI activity and t-PA antigen with peak values at 0600 in all three groups. Mean (SEM) PAI activity (IU/ml) at 2200, 0600, and 1400 was 6.1 (1.1), 11.0 (1.3), and 4.4 (0.6) in the variant angina group; 1.8 (0.7), 5.6 (1.1), and 1.2 (0.3) in the stable exertional angina group; and 1.1 (0.5), 4.5 (0.8), and 0.7 (0.3) in the control group. Furthermore, both plasma PAI activity and t-PA antigen concentrations were significantly higher in the variant angina group than in the stable exertional angina group and the control group at each sampling time. In patients with variant angina there was a circadian variation in fibrinolytic activity, which was lowest in the early morning, and impaired fibrinolytic activity particularly in the early morning, when attacks of angina occur most frequently.


Atopic dermatitis and ascariasis in children aged 2 to 10 years.
To assess the association between atopic dermatitis (AD) and ascariasis in 2 to 10-aged children from the neighborhood Pedregal, in the city of Campina Grande, Brazil, an area of low socioeconomic index. Cross-sectional study conducted with the use of the standard questionnaire from the International Study of Asthma and Allergies in Childhood (ISAAC) and stool parasitological exam for Ascaris lumbricoides. The dependent variable was AD diagnosis: absent, mild, and severe. Multivariate logistic regression and descriptive analysis of the variable were used. Associations were estimated using relative risk (RR) and odds ratio (OR). The statistical inference was based on 95% confidence intervals (95%CI). We assessed 1,195 children, 612 (51.2%) female. The AD prevalence was 24.6%, and ascariasis prevalence was 26.1%. In the mild AD group of children, 44 (36.7%) were infected by A. lumbricoides while in the severe AD group, 40 (22.9%) had the same geohelminthosis (p = 0.01). Comparing negative cases of AD between mild and severe forms, the infection with A. lumbricoides increased the frequency of mild AD (RR = 1.7; p = 0.009), but not the severe form (RR = 0.86; p = 0.46). Evaluating only the positive cases of dermatitis, 120 mild AD (40.8%) and 176 with severe AD (58.2%), it can be said that the exposure to the parasite decreased the frequency of the severe form of dermatitis (RR = 1.46; p = 0.016). There is a high prevalence of AD and of ascariasis in the population studied. The severe AD is related to lower parasitemia of A. lumbricoides.


From grief, guilt pain and stigma to hope and pride - a systematic review and meta-analysis of mixed-method research of the psychosocial impact of stillbirth.
Despite improvements in maternity healthcare services over the last few decades, more than 2.7 million babies worldwide are stillborn each year. The global health agenda is silent about stillbirth, perhaps, in part, because its wider impact has not been systematically analysed or understood before now across the world. Our study aimed to systematically review, evaluate and summarise the current evidence regarding the psychosocial impact of stillbirth to parents and their families, with the aim of improving guidance in bereavement care worldwide. Systematic review and meta-summary (quantitative aggregation of qualitative findings) of quantitative, qualitative, and mixed-methods studies. All languages and countries were included. Two thousand, six hundred and nineteen abstracts were identified; 144 studies were included. Frequency effect sizes (FES %) were calculated for each theme, as a measure of their prevalence in the literature. Themes ranged from negative psychological symptoms post bereavement (77 · 1) and in subsequent pregnancies (27 · 1), to disenfranchised grief (31 · 2), and incongruent grief (28 · 5), There was also impact on siblings (23 · 6) and on the wider family (2 · 8). They included mixed-feelings about decisions made when the baby died (12 · 5), avoidance of memories (13 · 2), anxiety over other children (7 · 6), chronic pain and fatigue (6 · 9), and a different approach to the use of healthcare services (6 · 9). Some themes were particularly prominent in studies of fathers; grief suppression (avoidance)(18 · 1), employment difficulties, financial debt (5 · 6), and increased substance use (4 · 2). Others found in studies specific to mothers included altered body image (3 · 5) and impact on quality of life (2 · 1). Counter-intuitively, Some themes had mixed connotations. These included parental pride in the baby (5 · 6), motivation for engagement in healthcare improvement (4 · 2) and changed approaches to life and death, self-esteem, and own identity (25 · 7). In studies from low/middle income countries, stigmatisation (13 · 2) and pressure to prioritise or delay conception (9) were especially prevalent. Experiencing the birth of a stillborn child is a life-changing event. The focus of the consequences may vary with parent gender and country. Stillbirth can have devastating psychological, physical and social costs, with ongoing effects on interpersonal relationships and subsequently born children. However, parents who experience the tragedy of stillbirth can develop resilience and new life-skills and capacities. Future research should focus on developing interventions that may reduce the psychosocial cost of stillbirth.


Canadian Association of Radiologists White Paper on De-Identification of Medical Imaging: Part 1, General Principles.
The application of big data, radiomics, machine learning, and artificial intelligence (AI) algorithms in radiology requires access to large data sets containing personal health information. Because machine learning projects often require collaboration between different sites or data transfer to a third party, precautions are required to safeguard patient privacy. Safety measures are required to prevent inadvertent access to and transfer of identifiable information. The Canadian Association of Radiologists (CAR) is the national voice of radiology committed to promoting the highest standards in patient-centered imaging, lifelong learning, and research. The CAR has created an AI Ethical and Legal standing committee with the mandate to guide the medical imaging community in terms of best practices in data management, access to health care data, de-identification, and accountability practices. Part 1 of this article will inform CAR members on principles of de-identification, pseudonymization, encryption, direct and indirect identifiers, k-anonymization, risks of reidentification, implementations, data set release models, and validation of AI algorithms, with a view to developing appropriate standards to safeguard patient information effectively.


Expression on porcine gamma delta lymphocytes of a phylogenetically conserved surface antigen previously restricted in expression to ruminant gamma delta T lymphocytes.
A 180,000 MW molecule has been identified on porcine leucocytes that is the homologue of the 215,000/300,000 MW WC1 (T19) leucocyte antigen previously considered to be restricted to ruminants. In ruminants the WC1 molecule is expressed by a T-cell subpopulation that is CD2-CD4-CD8-CD5+ and that is gamma delta T-cell receptor positive (TcR+). In pigs, the 180,000 MW molecule, identified by a new monoclonal antibody CC101, is expressed by a gamma delta TcR+ T-cell subpopulation that is also CD2-CD4-CD8-. The p180+ cells are a major T-cell subpopulation comprising approximately 40% of the peripheral blood mononuclear cells from 6-9-month-old pigs. Expression of p180 identifies the majority of the CD2-CD4-CD8- T cells in porcine blood. The p180+ T cells have a distribution in lymphoid tissues that is distinct from that of T cells that express the CD2, CD4 or CD8 molecules. They are evident particularly in the thymic medulla, the epithelium, lamina propria and interfollicular areas of the small intestine, and the superficial dermis of the skin, but largely absent from conventional T-dependent areas of secondary lymphoid tissue.


Pentobarbital coma for refractory intra-cranial hypertension after severe traumatic brain injury: mortality predictions and one-year outcomes in 55 patients.
To identify predictors of mortality and long-term outcomes in survivors after pentobarbital coma (PBC) in patients failing current treatment standards for severe traumatic brain injuries (TBI). This is a retrospective cohort study of severe TBI patients receiving PBC at Level I Trauma Center and tertiary university hospital. Four thousand nine hundred thirty-four patients were admitted to the trauma intensive care unit with severe TBI (head Abbreviated Injury Scale >or= 3) between April 1998 and December 2004. Six hundred eleven received intracranial pressure (ICP) monitoring and 58 received PBC. Three patients underwent craniotomy for intracranial mass lesion and were excluded. The study group received standardized medical management for severe TBI including opiates, benzodiazepines, elevation of the head of bed, avoidance of hypotension and hypercapnia and hyperosmolar therapy (HOsmRx). In addition, 31 of 55 patients (56%) underwent placement of intraventricular catheters for cerebrospinal fluid drainage. If routine medical management and cerebrospinal fluid diversion failed to control ICP, then the patient was determined to have refractory intracranial hypertension (RICH) and PBC treatment was initiated. PBC was performed with pentobarbital infusion with continuous electroencephalogram monitoring to ensure adequate burst suppression. The measurements include serum sodium (Na) and osmolality (Osm) were assessed as indicators for initiation of PBC and to estimate the 50% mortality cut-points when controlling for ICP. Follow-up functional outcomes were assessed using the Glasgow Outcome Scale and stratified according to admission Glasgow Coma Scale score and Marshall computed tomography classification. Of the 55 PBC patients, 22 (40%) survived at discharge. 19 of 22 had long-term follow-up (1 year or more) available. Of these, 13 (68%) were normal or functionally independent (Glasgow Outcome Scale score 4 or 5). Serum Na and Osm were associated with death (p < 0.05) when controlling for ICP. The 50% mortality cut-points were Na of 160 mEq/L and Osm of 330 mOsm/kg H2O. Median minimum cerebral perfusion pressure after PBC was 42 mm Hg in survivors and 34 mm Hg in nonsurvivors (p = 0.013). In patients with severe TBI and RICH, survival at discharge of 40% with good functional outcomes in 68% of survivors at 1 year or more can be achieved with PBC after failure of HOsmRx. Based on 50% mortality cut-points, analysis suggests the limits of HOsmRx to be Na of 160 mEq/L and Osm of 330 mOsm/Kg H2O. Maintenance of higher cerebral perfusion pressure after PBC is associated with survival. PBC treatment of RIH may be even more important when other treatments of RIH, such as decompressive craniectomy, are not available.


Evaluation of chemopreventive potential of Strobilanthes crispus against colon cancer formation in vitro and in vivo.
With cancer being one of the major causes of death around the world, studies are ongoing to find new chemotherapeutic leads. There are common mechanisms for colorectal cancer (CRC) formation. Several are connected with oxidative stress-induced cell apoptosis and others are related to imbalanced homeostasis or intake of drugs/toxins. Plants that have been used for decades in folk and traditional medicine have been accepted as one of the commonest sources of discovered natural agents of cancer chemotherapy and chemoprevention. The aim was to study the antioxidant and chemopreventive effects of Strobilanthes crispus on colorectal cancer formation. Five groups of rats were injected subcutaneously with AOM, 15 mg/kg body weight, each once weekly for 2 weeks. The cancer group was continued on 10 % Tween-20 feeding for 8 weeks. The standard drug group was continued on 35 mg/kg 5-fluorouracil intraperitoneal injection twice a week for 8 weeks, and the experimental groups were continued on 250 and 500 mg/kg S. crispus extract oral feeding for 8 weeks, respectively. The normal group was injected subcutaneously with normal saline once a week for 2 weeks, followed by oral administration of 10 % Tween-20 for 8 weeks. All the rats were sacrificed after 10 weeks. The colons were evaluated grossly and histopathologically for aberrant crypt foci (ACF). Gene expression was performed for Bax, Bcl2, Defa24, Slc24a3, and APC genes by real-time PCR. S. crispus and its fractions were evaluated for their chemopreventive effects against human colorectal adenocarcinoma cell line HT29 and cytotoxicity for normal human colon epithelial cell line CCD 841, and the active fraction was assessed for its components. We observed significant decrease in total colonic ACF formation, malonaldehyde (MDA) and lactate dehydrogenase (LDH), increase in superoxide dismutase (SOD), up-regulation of APC, Bax and Slc24a3, and down-regulation of Defa24 and Bcl-2 in rats treated with Strobilanthes crispus. Our results support the in vivo protection of S. crispus against CRC formation (azoxymethane-induced aberrant crypt foci) and suggest that the mechanism is highly specific to protect from oxidative insults and the following apoptotic cascade.


Anti-fouling strategies for central venous catheters.
Central venous catheters (CVCs) are ubiquitous in the healthcare industry and carry two common complications, catheter related infections and occlusion, particularly by thrombus. Catheter-related bloodstream infections (CRBSI) are an important cause of nosocomial infections that increase patient morbidity, mortality, and hospital cost. Innovative design strategies for intravenous catheters can help reduce these preventable infections. Antimicrobial coatings can play a major role in preventing disease. These coatings can be divided into two major categories: drug eluting and non-drug eluting. Much of these catheter designs are targeted at preventing the formation of microbial biofilms that make treatment of CRBSI nearly impossible without removal of the intravenous device. Exciting developments in catheter impregnation with antibiotics as well as nanoscale surface design promise innovative changes in the way that physicians manage intravenous catheters. Occlusion of a catheter renders the catheter unusable and is often treated by tissue plasminogen activator administration or replacement of the line. Prevention of this complication requires a thorough understanding of the mechanisms of platelet aggregation, signaling and cross-linking. This article will look at the advances in biomaterial design specifically drug eluting, non-drug eluting, lubricious coatings and micropatterning as well as some of the characteristics of each as they relate to CVCs.


The evolutionary paradox and the missing heritability of schizophrenia.
Schizophrenia is one of the most detrimental common psychiatric disorders, occurring at a prevalence of approximately 1%, and characterized by increased mortality and reduced reproduction, especially in men. The heritability has been estimated around 70% and the genome-wide association meta-analyses conducted by the Psychiatric Genomics Consortium have been successful at identifying an increasing number of risk loci. Various theories have been proposed to explain why genetic variants that predispose to schizophrenia persist in the population, despite the fitness reduction in affected individuals, a question known as the evolutionary paradox. In this review, we consider evolutionary perspectives of schizophrenia and of the empirical evidence that may support these perspectives. Proposed evolutionary explanations include balancing selection, fitness trade-offs, fluctuating environments, sexual selection, mutation-selection balance and genomic conflicts. We address the expectations about the genetic architecture of schizophrenia that are predicted by different evolutionary scenarios and discuss the implications for genetic studies. Several potential sources of "missing" heritability, including gene-environment interactions, epigenetic variation, and rare genetic variation are examined from an evolutionary perspective. A better understanding of evolutionary history may provide valuable clues to the genetic architecture of schizophrenia and other psychiatric disorders, which is highly relevant to genetic studies that aim to detect genetic risk variants.


The incidence of dementia in England and Wales: findings from the five identical sites of the MRC CFA Study.
Although incidence of dementia is known to vary between nations, variation within country has not been explored because most incidence studies are single site or have insufficient numbers to compare sites. Few countries have conducted multisite incidence studies in order to facilitate national comparisons. This study aims to provide robust measures of the variation of the incidence of dementia across sites within England and Wales and produce overall estimates by age and sex. The Medical Research Council Cognitive Function and Ageing Study used identical methodology in five diverse sites across the United Kingdom, each with different risk patterns and mortality rates. Incidence has been estimated using likelihood-based methods between the first two waves of interviews. Incidence rates rise with age, particularly above the age of 75 y, from 6.7 [corrected] (95% confidence interval, 3.8 [corrected]-12.4 [corrected]) per 1,000 person years at age 65-69 y to 68.5 [corrected] (95% confidence interval, 52.5 [corrected]-88.1 [corrected]) per 1,000 person years at age 85 y and above. The rate of increase for both sexes is marked, and continues into the oldest age groups. Hence, it is estimated that approximately 163,000 [corrected] new cases of dementia occur in England and Wales each year. There is no convincing evidence of variation across sites, and incidence rates do not reflect the variations in the prevalence of possible risk factors in these sites. There is no evidence, within England and Wales, of variation in dementia incidence across sites. Dementia incidence rates do not tail off at the oldest ages.


2-Arylindenes and 2-arylindenones: molecular structures and considerations in the binding orientation of unsymmetrical nonsteroidal ligands to the estrogen receptor.
We have studied how 2-arylindene systems, unsymmetrical nonsteroidal estrogens, orient themselves within the binding site of the estrogen receptor, relative to estradiol, by making a comprehensive comparison of the binding affinity of 16 analogues. These analogues are representatives of two major classes, those substituted at C-3 with an ethyl or with a phenyl substituent; within each class there are members that have different patterns of hydroxyl group substitution and C-1 oxo or alkyl substitution. Orientational preferences were inferred from the relative binding affinities and were supplemented by computer graphic molecular overlap studies that utilized crystal structures of selected representative compounds and the known tolerance of the estrogen receptor to substituents on the steroidal ligand estradiol. 2-Arylindenes with a 3-aryl substituent appear to orient with the indene system mimicking the A- and B-rings of estradiol (indene/AB mode). This orientation is supported by the fact that hydroxyl substitution at C-6 in the indene markedly elevates binding relative to hydroxyl substitution at the para position of the 2-phenyl substituent. A C-1 oxo substituent increases binding further, but a C-1 alkyl group has little effect. By contrast, the 2-arylindenes with a C-3 ethyl substituent appear to bind with the pendant C-2 ring, mimicking the A-ring of estradiol (pendant/A mode), as hydroxyl substitution in this ring elevates binding relative to the C-6 hydroxy analogues. C-1 alkyl substitution elevates binding affinity in this series; such a substituent in a C-1 S configuration would be projected into the receptor region normally occupied by the high-affinity 7 alpha- or 11 beta-alkyl estradiols. A C-1 oxo substituent produces only a modest binding enhancement in the C-3 ethyl series. A thermodynamic evaluation of receptor fit suggests that the smaller 3-ethyl-2-arylindenes are more efficient than the 2,3-diarylindenes in the use of the molecular bulk to achieve receptor binding. This analysis of the orientational preference of 2-arylindene nonsteroidal estrogens has important implications in the design of donor/acceptor-substituted 2-arylindenes as fluorescent ligands for the estrogen receptor.


Effect of 12 weeks of yoga training on the somatization, psychological symptoms, and stress-related biomarkers of healthy women.
Previous studies have shown that the practice of yoga reduces perceived stress and negative feelings and that it improves psychological symptoms. Our previous study also suggested that long-term yoga training improves stress-related psychological symptoms such as anxiety and anger. However, little is known about the beneficial effects of yoga practice on somatization, the most common stress-related physical symptoms, and stress-related biomarkers. We performed a prospective, single arm study to examine the beneficial effects of 12 weeks of yoga training on somatization, psychological symptoms, and stress-related biomarkers. We recruited healthy women who had no experience with yoga. The data of 24 participants who were followed during 12 weeks of yoga training were analyzed. Somatization and psychological symptoms were assessed before and after 12 weeks of yoga training using the Profile of Mood State (POMS) and the Symptom Checklist-90-Revised (SCL-90-R) questionnaires. Urinary 8-hydroxydeoxyguanosine (8-OHdG), biopyrrin, and cortisol levels were measured as stress-related biomarkers. The Wilcoxon signed-rank test was used to compare the stress-related biomarkers and the scores of questionnaires before and after 12 weeks of yoga training. After 12 weeks of yoga training, all negative subscale scores (tension-anxiety, depression, anger-hostility, fatigue, and confusion) from the POMS and somatization, anxiety, depression, and hostility from the SCL-90-R were significantly decreased compared with those before starting yoga training. Contrary to our expectation, the urinary 8-OHdG concentration after 12 weeks of yoga training showed a significant increase compared with that before starting yoga training. No significant changes were observed in the levels of urinary biopyrrin and cortisol after the 12 weeks of yoga training. Yoga training has the potential to reduce the somatization score and the scores related to mental health indicators, such as anxiety, depression, anger, and fatigue. The present findings suggest that yoga can improve somatization and mental health status and has implications for the prevention of psychosomatic symptoms in healthy women. University Hospital Medical Information Network (UMIN CTR) UMIN000007868.


ARIA 2016: Care pathways implementing emerging technologies for predictive medicine in rhinitis and asthma across the life cycle.
The Allergic Rhinitis and its Impact on Asthma (ARIA) initiative commenced during a World Health Organization workshop in 1999. The initial goals were (1) to propose a new allergic rhinitis classification, (2) to promote the concept of multi-morbidity in asthma and rhinitis and (3) to develop guidelines with all stakeholders that could be used globally for all countries and populations. ARIA-disseminated and implemented in over 70 countries globally-is now focusing on the implementation of emerging technologies for individualized and predictive medicine. MASK [MACVIA (Contre les Maladies Chroniques pour un Vieillissement Actif)-ARIA Sentinel NetworK] uses mobile technology to develop care pathways for the management of rhinitis and asthma by a multi-disciplinary group and by patients themselves. An app (Android and iOS) is available in 20 countries and 15 languages. It uses a visual analogue scale to assess symptom control and work productivity as well as a clinical decision support system. It is associated with an inter-operable tablet for physicians and other health care professionals. The scaling up strategy uses the recommendations of the European Innovation Partnership on Active and Healthy Ageing. The aim of the novel ARIA approach is to provide an active and healthy life to rhinitis sufferers, whatever their age, sex or socio-economic status, in order to reduce health and social inequalities incurred by the disease.


Phylogeographic evidence of crop neodiversity in sorghum.
Sorghum has shown the adaptability necessary to sustain its improvement during time and geographical extension despite a genetic foundation constricted by domestication bottlenecks. Initially domesticated in the northeastern part of sub-Saharan Africa several millenia ago, sorghum quickly spread throughout Africa, and to Asia. We performed phylogeographic analysis of sequence diversity for six candidate genes for grain quality (Shrunken2, Brittle2, Soluble starch synthaseI, Waxy, Amylose extender1, and Opaque2) in a representative sample of sorghum cultivars. Haplotypes along 1-kb segments appeared little affected by recombination. Sequence similarity enabled clustering of closely related alleles and discrimination of two or three distantly related groups depending on the gene. This scheme indicated that sorghum domestication involved structured founder populations, while confirming a specific status for the guinea margaritiferum subrace. Allele rooted genealogy revealed derivation relationships by mutation or, less frequently, by recombination. Comparison of germplasm compartments revealed contrasts between genes. Sh2, Bt2, and SssI displayed a loss of diversity outside the area of origin of sorghum, whereas O2 and, to some extent, Wx and Ae1 displayed novel variation, derived from postdomestication mutations. These are likely to have been conserved under the effect of human selection, thus releasing valuable neodiversity whose extent will influence germplasm management strategies.


The first α-helical domain of the vesicle-inducing protein in plastids 1 promotes oligomerization and lipid binding.
The vesicle-inducing protein in plastids 1 (Vipp1) is an essential component for thylakoid biogenesis in cyanobacteria and chloroplasts. Vipp1 proteins share significant structural similarity with their evolutionary ancestor PspA (bacterial phage shock protein A), namely a predominantly α-helical structure, the formation of oligomeric high molecular weight complexes (HMW-Cs) and a tight association with membranes. Here, we elucidated domains of Vipp1 from Arabidopsis thaliana involved in homo-oligomerization as well as association with chloroplast inner envelope membranes. We could show that the 21 N-terminal amino acids of Vipp1, which form the first α-helix of the protein, are essential for assembly of the 2 MDa HMW-C but are not needed for formation of smaller subcomplexes. Interestingly, removal of this domain also interferes with association of the Vipp1 protein to the inner envelope. Fourier transform infrared spectroscopy of recombinant Vipp1 further indicates that Escherichia coli lipids bind tightly enough that they can be co-purified with the protein. This feature also depends on the presence of the first helix, which strongly supports an interaction of lipids with the Vipp1 HMW-C but not with smaller subcomplexes. Therefore, Vipp1 oligomerization appears to be a prerequisite for its membrane association. Our results further highlight structural differences between Vipp1 and PspA, which might be important in regard to their different function in thylakoid biogenesis and bacterial stress response, respectively.


Task shifting for emergency obstetric surgery in district hospitals in Senegal.
Due to a long-term shortage of obstetricians, the Ministry of Health of Senegal and Dakar University Obstetric Department agreed in 1998 to train district teams consisting of an anaesthetist, general practitioner and surgical assistant in emergency obstetric surgery. An evaluation of the policy was carried out in three districts in 2006, covering trends in rates of major obstetric interventions, outcomes in newborns and mothers, and the views of key informants, community members and final year medical students. From 2001 to 2006, 11 surgical teams were trained but only six were functioning in 2006. The current rate of training is not rapid enough to cover all districts by 2015. An increase in the rate of interventions was noted as soon as a team had been put in place, but unmet need persisted. Central decision-makers considered the policy more viable than training gynaecologists for district hospitals, but resistance from senior academic clinicians, a perceived lack of career progression among the doctors trained, and lack of programme coordination were obstacles. Practitioners felt the work was valuable, but complained of low additional pay and not being replaced during training. Communities appreciated that the services saved lives and money, but called for improved information and greater continuity of care.


The epidemiology of injury in ATV and motocross sports.
Off-road motorsports are popular in rural and suburban areas, and allow for racing, recreation, and easy access to backcountry destinations. This chapter will review the incidence and types of injuries sustained in off-road motorsports. We completed a structured review of motocross and all-terrain vehicle (ATV) injuries, assessing for injury rates, risk factors, and mortality figures. Information for this study was obtained from a PubMed search under the terms 'motocross', 'motorcross', 'all-terrain vehicles', 'injury', 'motorcycle', 'ATV'. Abstracts and articles in the English language from 1980 onward were reviewed. Further statistics were obtained from the US Consumer Product Safety Commission publications. Operating vehicles off-road requires coordination, experience, and training. Motocross is an organized sport with national associations governing the competition of highly trained athletes. ATVs are used both recreationally and commercially, typically for farming and ranching. ATV use appears more dangerous that motocross, with a higher mortality rate, disproportionately for children. Both sports continue to have high rates of head, spinal cord, and extremity injury. Future prospective studies in off-road motorsports should evaluate the risk factors for injury and target specific areas for injury prevention. Improved training programs, use and improvement of safety helmets, and for ATV use, limiting access to minors, may improve the overall safety of off-road motorsports.


Sorption, desorption, and surface oxidative fate of nicotine.
Nicotine dynamics in an indoor environment can be greatly affected by building parameters (e.g. relative humidity (RH), air exchange rate (AER), and presence of ozone), as well as surface parameters (e.g. surface area (SA) and polarity). To better understand the indoor fate of nicotine, these parameter effects on its sorption, desorption, and oxidation rates were investigated on model indoor surfaces that included fabrics, wallboard paper, and wood materials. Nicotine sorption under dry conditions was enhanced by higher SA and higher polarity of the substrate. Interestingly, nicotine sorption to cotton and nylon was facilitated by increased RH, while sorption to polyester was hindered by it. Desorption was affected by RH, AER, and surface type. Heterogeneous nicotine-ozone reaction was investigated by Fourier transform infrared spectrometry with attenuated total reflection (FTIR-ATR), and revealed a pseudo first-order surface reaction rate of 0.035 +/- 0.015 min(-1) (at [O(3)] = 6 +/- 0.3 x 10(15) molecules cm(-3)) that was partially inhibited at high RH. Extrapolation to a lower ozone level ([O(3)] = 42 ppb) showed oxidation on the order of 10(-5) min(-1) corresponding to a half-life of 1 week. In addition, similar surface products were identified in dry and high RH using gas chromatography-mass spectrometry (GC-MS). However, FTIR analysis revealed different product spectra for these conditions, suggesting additional unidentified products and association with surface water. Knowing the indoor fate of condensed and gas phase nicotine and its oxidation products will provide a better understanding of nicotine's impact on personal exposures as well as overall indoor air quality.


Evaluation of non-inferiority of intradermal versus adjuvanted seasonal influenza vaccine using two serological techniques: a randomised comparative study.
Although seasonal influenza vaccine is effective in the elderly, immune responses to vaccination are lower in the elderly than in younger adults. Strategies to optimise responses to vaccination in the elderly include using an adjuvanted vaccine or using an intradermal vaccination route. The immunogenicity of an intradermal seasonal influenza vaccine was compared with that of an adjuvanted vaccine in the elderly. Elderly volunteers (age > or = 65 years) were randomised to receive a single dose of trivalent seasonal influenza vaccine: either a split-virion vaccine containing 15 microg haemagglutinin [HA]/strain/0.1-ml dose administered intradermally, or a subunit vaccine (15 microg HA/strain/0.5-ml dose) adjuvanted with MF59C.1 and administered intramuscularly. Blood samples were taken before and 21 +/- 3 days post-vaccination. Anti-HA antibody titres were assessed using haemagglutination inhibition (HI) and single radial haemolysis (SRH) methods. We aimed to show that the intradermal vaccine was non-inferior to the adjuvanted vaccine. A total of 795 participants were enrolled (intradermal vaccine n = 398; adjuvanted vaccine n = 397). Non-inferiority of the intradermal vaccine was demonstrated for the A/H1N1 and B strains, but not for the A/H3N2 strain (upper bound of the 95% CI = 1.53) using the HI method, and for all three strains by the SRH method. A post-hoc analysis of covariance to adjust for baseline antibody titres demonstrated the non-inferiority of the intradermal vaccine by HI and SRH methods for all three strains. Both vaccines were, in general, well tolerated; the incidence of injection-site reactions was higher for the intradermal (70.1%) than the adjuvanted vaccine (33.8%) but these reactions were mild and of short duration. The immunogenicity and safety of the intradermal seasonal influenza vaccine in the elderly was comparable with that of the adjuvanted vaccine. Intradermal vaccination to target the immune properties of the skin appears to be an appropriate strategy to address the challenge of declining immune responses in the elderly. ClinicalTrials.gov: NCT00554333.


Identification of New Pathogenic Races of Common Bunt and Dwarf Bunt Fungi, and Evaluation of Known Races Using an Expanded Set of Differential Wheat Lines.
Pathogenic races of Tilletia caries and T. foetida, which cause common bunt of wheat (Triticum aestivum), and Tilletia contraversa, which causes dwarf bunt of wheat, have been identified previously by their reaction to 10 differential wheat lines, each containing single bunt resistance genes Bt1 through Bt10. The reactions of races to the differential wheat lines follow the classic gene-for gene system for host-pathogen interactions. The pathogens are closely related and resistance to both diseases in wheat is controlled by the same genes. To better define pathogenic races, six additional wheat lines containing the genes Bt11 through Bt15 and a wheat line with a resistance factor designated as Btp were added to the set of 10 differentials and tested with all named U.S. races of common bunt and dwarf bunt. In addition, new isolates of dwarf bunt, and common bunt from hybrids and field collections, were tested with all 16 differentials for race identification. Six new races of T. caries, five new races of T. foetida, and two new races of T. contraversa were identified. Races of common bunt virulent to Bt8 or Bt12, and dwarf bunt races virulent to the combinations of Bt11 and Bt12, and Bt8, Bt9, Bt10, Bt11, and Bt12, were identified for the first time. Comparison of the reactions of the common bunt races with the Bt14 and Bt15 differentials grown in different environments after initial infection showed that these genes are temperature sensitive, indicating they should be excluded from the set of differential lines to avoid ambiguity in determining virulent or avirulent reactions. In the previous list of bunt races, there were races that had the same reaction to the set of 10 differentials but were designated as different races. These races were not differentiated further with the six additional differentials, indicating that the duplicate races should be dropped from the list of pathogenic races. The new races of common bunt and dwarf bunt identified have unique patterns of virulence that allow specific targeting and elucidation of bunt resistance genes in wheat and will aid the development of bunt-resistant wheat cultivars.


A time for restraint.
The debate on the use of human embryos for research will be one of the more important issues of the 21st century. Unlike recombinant DNA technology, embryonic stem cell research most probably will result in the destruction of living embryos. Many people consider this research immoral, illegal, and unnecessary. Therefore, it is imperative to proceed cautiously. Federal funding of research using human embryos or pluripotent cells derived from them would be inappropriate until further resolution of the ethical issues has been achieved.


Purification and functional characterization of type II DNA topoisomerase from rat testis and comparison with topoisomerase II from liver.
A number of studies in yeast have shown that DNA topoisomerase II is essential for chromosome condensation and disjunction during mitosis at the metaphase/anaphase transition and meiosis I. Accordingly, kinetic and mechanistic studies have implied a role for topoisomerase II in chromosome disjunction. As a step toward understanding the nature and role of topoisomerase II in a mammalian germline in vivo, we have purified topoisomerase II from rat testis to homogeneity and ascertained several of its catalytic activities in conjunction with that of the purified enzyme from liver. The purified enzymes appeared to be monomers under denaturing conditions; however, they differed in their relative molecular mass. Topoisomerase II from testis and liver have apparent molecular masses of 150 +/- 10 kDa and 160 +/- 10 kDa, respectively. The native molecular mass of testis topoisomerase II as assayed by immunoblot analysis of cell-free extracts, prepared in the presence of SDS and a number of protease inhibitors, corroborated with the size of the purified enzyme. Both enzymes are able to promote decatenation and relax supercoiled DNA substrates in an ATP and Mg(2+)-dependent manner. However, quantitative comparison of catalytic properties of topoisomerase II from testis with that of the enzyme from liver displayed significant differences in their efficiencies. Optimal pH values for testis enzyme are 6.5 to 8.5 while they are 6 to 7.5 for the liver enzyme. Intriguingly, the relaxation activity of liver topoisomerase II was inhibited by potassium glutamate at 1 M, whereas testis enzyme required about half its concentration. These findings argue that topoisomerase II from rat testis is structurally distinct from that of its somatic form and the functional differences between the two enzymes parallels with the physiological environment that is unique to these two tissues.


Phosphorylation of BRN2 modulates its interaction with the Pax3 promoter to control melanocyte migration and proliferation.
MITF-M and PAX3 are proteins central to the establishment and transformation of the melanocyte lineage. They control various cellular mechanisms, including migration and proliferation. BRN2 is a POU domain transcription factor expressed in melanoma cell lines and is involved in proliferation and invasion, at least in part by regulating the expression of MITF-M and PAX3. The T361 and S362 residues of BRN2, both in the POU domain, are conserved throughout the POU protein family and are targets for phosphorylation, but their roles in vivo remain unknown. To examine the role of this phosphorylation, we generated mutant BRN2 in which these two residues were replaced with alanines (BRN2TS→BRN2AA). When expressed in melanocytes in vitro or in the melanocyte lineage in transgenic mice, BRN2TS induced proliferation and repressed migration, whereas BRN2AA repressed both proliferation and migration. BRN2TS and BRN2AA bound and repressed the MITF-M promoter, whereas PAX3 transcription was induced by BRN2TS but repressed by BRN2AA. Expression of the BRN2AA transgene in a Mitf heterozygous background and in a Pax3 mutant background enhanced the coat color phenotype. Our findings show that melanocyte migration and proliferation are controlled both through the regulation of PAX3 by nonphosphorylated BRN2 and through the regulation of MITF-M by the overall BRN2 level.


Hybrid integrate-and-fire model of a bursting neuron.
We present a reduction of a Hodgkin-Huxley (HH)--style bursting model to a hybridized integrate-and-fire (IF) formalism based on a thorough bifurcation analysis of the neuron's dynamics. The model incorporates HH--style equations to evolve the subthreshold currents and includes IF mechanisms to characterize spike events and mediate interactions between the subthreshold and spiking currents. The hybrid IF model successfully reproduces the dynamic behavior and temporal characteristics of the full model over a wide range of activity, including bursting and tonic firing. Comparisons of timed computer simulations of the reduced model and the original model for both single neurons and moderately sized networks (n < or = 500) show that this model offers improvement in computational speed over the HH--style bursting model.


Health and Taxes: Hospitals, Community Health and the IRS.
The Affordable Care Act created new conditions of federal tax exemption for nonprofit hospitals, including a requirement that hospitals conduct a community health needs assessment (CHNA) every three years to identify significant health needs in their communities and then develop and implement a strategy responding to those needs. As a result, hospitals must now do more than provide charity care to their patients in exchange for the benefits of tax exemption. The CHNA requirement has the potential both to prompt a radical change in hospitals' relationship to their communities and to enlist hospitals as meaningful contributors to community health improvement initiatives. Final regulations issued in December 2014 clarify hospitals' obligations under the CHNA requirement, but could do more to facilitate hospitals' engagement in collaborative community health projects. The Internal Revenue Service (IRS) has a rich opportunity, while hospitals are still learning to conduct CHNAs, to develop guidance establishing clear but flexible expectations for how providers should assess and address community needs. This Article urges the IRS to seize that opportunity by refining its regulatory framework for the CHNA requirement. Specifically, the IRS should more robustly promote transparency, accountability, community engagement, and collaboration while simultaneously leaving hospitals a good degree of flexibility. By promoting alignment between hospitals' regulatory compliance activities and broader community health improvement initiatives, the IRS could play a meaningful role in efforts to reorient our system towards promoting health and not simply treating illness.


Determinants of the choice of GP practice registration in England: evidence from a discrete choice experiment.
There have been growing concerns that general practitioner (GP) services in England, which are based on registration with a single practice located near the patient's home, are not sufficiently convenient for patients. To inform the decision as to whether to change registration rules allowing patients to register 'out-of-area' and to estimate the demand for this wider choice, we undertook a discrete choice experiment with 1706 respondents. Latent class models were used to analyse preferences for GP practice registration comparing preferences for neighbourhood and non-neighbourhood practices. We find that there is some appetite for registering outside the neighbourhood, but this preference is not uniformly shared across the population. Specifically individuals who are less mobile (e.g. older people and those with caring responsibilities), or satisfied with their local practice are less likely to be interested in registering at a practice outside their neighbourhood. Overall, people feel most strongly about obtaining an appointment with a GP as quickly as possible. Respondents regarded weekend opening as less important than other factors, and particularly less important than extended practice opening hours from Monday to Friday. Assuming a constant demand for GP services, a policy encouraging GP practices to extend their opening hours during the week is likely to decrease the average patient waiting time for an appointment and is likely to be preferred by patients.


Pan-cancer characterisation of microRNA across cancer hallmarks reveals microRNA-mediated downregulation of tumour suppressors.
microRNAs are key regulators of the human transcriptome across a number of diverse biological processes, such as development, aging and cancer, where particular miRNAs have been identified as tumour suppressive and oncogenic. In this work, we elucidate, in a comprehensive manner, across 15 epithelial cancer types comprising 7316 clinical samples from the Cancer Genome Atlas, the association of miRNA expression and target regulation with the phenotypic hallmarks of cancer. Utilising penalised regression techniques to integrate transcriptomic, methylation and mutation data, we find evidence for a complex map of interactions underlying the relationship of miRNA regulation and the hallmarks of cancer. This highlighted high redundancy for the oncomiR-1 cluster of oncogenic miRNAs, in particular hsa-miR-17-5p. In addition, we reveal extensive miRNA regulation of tumour suppressor genes such as PTEN, FAT4 and CDK12, uncovering an alternative mechanism of repression in the absence of mutation, methylation or copy number changes.


Trauma and post-traumatic stress disorder in women: a review.
In the aftermath of traumas such as combat or sexual assault, both men and women may experience similar symptoms, including post-traumatic stress disorder (PTSD). However, epidemiological studies have yielded higher rates of PTSD in women than in men in general populations, and there are also a number of gender differences in clinical presentation after trauma. Thus, in a study by the authors of patients presenting with physical trauma after interpersonal violence, women were more likely than men to have been previously assaulted, or to have sustained injury by a relative or someone known to them, but less likely to have used substances at the time of the assault or to require emergency surgery. A better understanding of the particular factors that contribute to higher rates of PTSD in women may ultimately shed light on the pathogenesis of this complex disorder. This article reviews gender differences in exposure to trauma and subsequent PTSD, emphasizing those features that characterize trauma and PTSD in women.


Psychophysical study of the visual sun location in pictures of cloudy and twilight skies inspired by Viking navigation.
In the late 1960s it was hypothesized that Vikings had been able to navigate the open seas, even when the sun was occluded by clouds or below the sea horizon, by using the angle of polarization of skylight. To detect the direction of skylight polarization, they were thought to have made use of birefringent crystals, called "sun-stones," and a large part of the scientific community still firmly believe that Vikings were capable of polarimetric navigation. However, there are some critics who treat the usefulness of skylight polarization for orientation under partly cloudy or twilight conditions with extreme skepticism. One of their counterarguments has been the assumption that solar positions or solar azimuth directions could be estimated quite accurately by the naked eye, even if the sun was behind clouds or below the sea horizon. Thus under partly cloudy or twilight conditions there might have been no serious need for a polarimetric method to determine the position of the sun. The aim of our study was to test quantitatively the validity of this qualitative counterargument. In our psychophysical laboratory experiments, test subjects were confronted with numerous 180 degrees field-of-view color photographs of partly cloudy skies with the sun occluded by clouds or of twilight skies with the sun below the horizon. The task of the subjects was to guess the position or the azimuth direction of the invisible sun with the naked eye. We calculated means and standard deviations of the estimated solar positions and azimuth angles to characterize the accuracy of the visual sun location. Our data do not support the common belief that the invisible sun can be located quite accurately from the celestial brightness and/or color patterns under cloudy or twilight conditions. Although our results underestimate the accuracy of visual sun location by experienced Viking navigators, the mentioned counterargument cannot be taken seriously as a valid criticism of the theory of the alleged polarimetric Viking navigation. Our results, however, do not bear on the polarimetric theory itself.


Functional multiple indicators, multiple causes measurement error models.
Objective measures of oxygen consumption and carbon dioxide production by mammals are used to predict their energy expenditure. Since energy expenditure is not directly observable, it can be viewed as a latent construct with multiple physical indirect measures such as respiratory quotient, volumetric oxygen consumption, and volumetric carbon dioxide production. Metabolic rate is defined as the rate at which metabolism occurs in the body. Metabolic rate is also not directly observable. However, heat is produced as a result of metabolic processes within the body. Therefore, metabolic rate can be approximated by heat production plus some errors. While energy expenditure and metabolic rates are correlated, they are not equivalent. Energy expenditure results from physical function, while metabolism can occur within the body without the occurrence of physical activities. In this manuscript, we present a novel approach for studying the relationship between metabolic rate and indicators of energy expenditure. We do so by extending our previous work on MIMIC ME models to allow responses that are sparsely observed functional data, defining the sparse functional multiple indicators, multiple cause measurement error (FMIMIC ME) models. The mean curves in our proposed methodology are modeled using basis splines. A novel approach for estimating the variance of the classical measurement error based on functional principal components is presented. The model parameters are estimated using the EM algorithm and a discussion of the model's identifiability is provided. We show that the defined model is not a trivial extension of longitudinal or functional data methods, due to the presence of the latent construct. Results from its application to data collected on Zucker diabetic fatty rats are provided. Simulation results investigating the properties of our approach are also presented.


Morphology of OLED Film Stacks Containing Solution-Processed Phosphorescent Dendrimers.
Organic light-emitting devices containing solution-processed emissive dendrimers can be highly efficient. The most efficient devices contain a blend of the light-emitting dendrimer in a host and one or more charge-transporting layers. Using neutron reflectometry measurements with in situ photoluminescence, we have investigated the structure of the as-formed film as well as the changes in film structure and dendrimer emission under thermal stress. It was found that the as-formed film stacks comprising poly(3,4-ethylenedioxythiophene):polystyrene sulfonate/host:dendrimer/1,3,5-tris(N-phenylbenzimidazol-2-yl)benzene (where the host was deuterated 4,4'-N,N'-di(carbazolyl)biphenyl or tris(4-carbazol-9-ylphenyl)amine, the host:dendrimer layer was solution-processed, and the 1,3,5-tris(N-phenylbenzimidazol-2-yl)benzene evaporated) had well-defined interfaces, indicating good wetting of each of the layers by the subsequently deposited layer. Upon thermal annealing, there was no change in the poly(3,4-ethylenedioxythiophene):polystyrene sulfonate/host:dendrimer interface, but once the temperature reached above the Tg of the host:dendrimer layer, it became a supercooled liquid into which 1,3,5-tris(N-phenylbenzimidazol-2-yl)benzene dissolved. When the film stacks were held at a temperature just above the onset of the diffusion process, they underwent an initial relatively fast diffusion process before reaching a quasi-stable state at that temperature.


Functional conservation of a forebrain enhancer from the elephant shark (Callorhinchus milii ) in zebrafish and mice.
The phylogenetic position of the elephant shark (Callorhinchus milii ) is particularly relevant to study the evolution of genes and gene regulation in vertebrates. Here we examine the evolution of Dlx homeobox gene regulation during vertebrate embryonic development with a particular focus on the forebrain. We first identified the elephant shark sequence orthologous to the URE2 cis -regulatory element of the mouse Dlx1/Dlx2 locus (herein named CmURE2). We then conducted a comparative study of the sequence and enhancer activity of CmURE2 with that of orthologous regulatory sequences from zebrafish and mouse. The CmURE2 sequence shows a high percentage of identity with its mouse and zebrafish counterparts but is overall more similar to mouse URE2 (MmURE2) than to zebrafish URE2 (DrURE2). In transgenic zebrafish and mouse embryos, CmURE2 displayed enhancer activity in the forebrain that overlapped with that of DrURE2 and MmURE2. However, we detected notable differences in the activity of the three sequences in the diencephalon. Outside of the forebrain, CmURE2 shows enhancer activity in areas such as the pharyngeal arches and dorsal root ganglia where its' counterparts are also active. Our transgenic assays show that part of the URE2 enhancer activity is conserved throughout jawed vertebrates but also that new characteristics have evolved in the different groups. Our study demonstrates that the elephant shark is a useful outgroup to study the evolution of regulatory mechanisms in vertebrates and to address how changes in the sequence of cis -regulatory elements translate into changes in their regulatory activity.


Are the stages of change relevant for the development and implementation of a web-based tailored alcohol intervention? A cross-sectional study.
Computer-tailored programs are a promising tool to stimulate health behavior change, such as reducing alcohol intake. Yet more research is needed to assess whether groups differing in their motivational level to change may need different types of feedback. Furthermore, it is unknown whether motivational level may also determine reactions to computer-tailored interventions. Our aim is to identify the potential relevance of the application of the stages of change concept in the development and implementation of alcohol interventions. A web-based instrument was used to disseminate a questionnaire and to provide tailored feedback messages among adults in the Netherlands (N = 170; 96 females). Motivational level was assessed by the stage of change construct. The survey furthermore assessed alcohol consumption, attitude, social influence, self-efficacy, and program evaluation (i.e., survey items, tailored advice, layout and functionality of the program). The Least Significant Difference method was used to compare people in different stages of change with regard to psychosocial determinants of drinking behavior and program evaluation. Of the respondents, 34.1% (n = 58) reported no intention to change to healthier drinking habits in the foreseeable future (precontemplation), 22.9% (n = 39) intended to improve their drinking behavior in the near future (contemplation/preparation) and 42.9% (n = 73) reported to currently adhere to the Dutch alcohol consumption guidelines (action/maintenance). When comparing the three groups, people in the action or maintenance stage reported the lowest number of pros of drinking alcohol, having most healthy drinking role models and the highest levels of self-efficacy regarding healthy drinking in difficult situations, whereas precontemplators reported to receive the least social support regarding healthy drinking. In general, the intervention was positively evaluated, but it seemed to be most appreciated by contemplators and preparers. Stage-matched interventions may be useful to encourage people to reduce their alcohol intake. Different factors seem to be important for people in different motivational stages. Longitudinal studies are needed to determine whether these factors also predict stage transition. The intervention could be optimized by tailoring the feedback messages more precisely to the needs of people in different motivational stages, for example by applying the different processes of change.


Irbesartan treatment does not influence plasma levels of the advanced glycation end products N(epsilon)(1-carboxymethyl)lysine and N(epsilon)(1-carboxyethyl)lysine in patients with type 2 diabetes and microalbuminuria. A randomized controlled trial.
In vitro and animal experiments have shown inhibiting effects of angiotensin receptor blockers (ARBs) on the formation of advanced glycation end products (AGEs), which are known to be involved in the development of cardiovascular complications in diabetes. However, sufficient human data to confirm such beneficial effects of ARBs on AGEs are lacking. Therefore, we investigated the effects of irbesartan treatment on plasma levels of the AGEs N(ε)(1-carboxymethyl)lysine (CML) and N(ε)(1-carboxyethyl)lysine (CEL) in hypertensive patients with type 2 diabetes and microalbuminuria. We analysed data from a multicentre, double-blind, parallel, randomized controlled trial in patients with type 2 diabetes and microalbuminuria, the primary goal of which was to examine the renoprotective effects of irbesartan treatment (150 or 300 mg daily). Secondary end points included plasma CML and CEL in the treatment arm receiving 300 mg irbesartan (n = 139) and in the placebo group (n = 125). Effects of treatment at 1- and 2-year follow-up were analysed by means of generalized estimating equations according to an intention-to-treat principle. Levels of CML and CEL did not differ between groups at baseline. No significant changes were observed in CML and CEL over time in either group and there was no effect of treatment on CML and CEL at any time-point. Mean differences for the irbesartan versus placebo group over time were -0.96 μmol/mol lysine (95% confidence interval: -3.43 to 1.51) for CML and -0.10 μmol/mol lysine (-0.76 to 0.56) for CEL. Long-term irbesartan treatment does not influence plasma levels of the AGE CML and CEL in patients with type 2 diabetes and microalbuminuria.


Interactive cueing with Walk-Mate for hemiparetic stroke rehabilitation.
Many techniques that compensate for locomotion problems in daily life using externally controlled stimulation have recently been reported. These techniques are beneficial for effortlessly supporting patients' locomotive functions, but the users of such devices must necessarily remain dependent on them. It is possible that some individuals with gait impairment may be prevented recovering locomotive function. From a rehabilitation viewpoint, it may therefore be supposed that ideally, devices that can be used in daily life to improve the locomotive functions of the body itself should be proposed. We evaluate the effectiveness of Walk-Mate, which has been used mainly as a gait compensation device, as a gait rehabilitation training device by analyzing improvement in locomotion before, during and after rehabilitation in hemiparetic patients and comparing it with a previous gait training method. Walk-Mate generates a model walking rhythm in response to a user's locomotion in real time, and by indicating this rhythm using auditory stimuli, provides a technology that supports walking by reducing asymmetries and fluctuations in foot contact rhythm. If patients can use the system to learn a regulated walking rhythm, then it may also be expected to fulfil the functions of a gait rehabilitation training device for daily life. With regard to asymmetry, significantly improvements were seen for compensatory movement during training using Walk-Mate, but improvements were not retained as rehabilitative results. Regarding fluctuations in the foot contact period, significant improvement was observed for compensatory movement during training and these significant improvements were retained as rehabilitative results. In addition, it became clear that such improvement could not be adequately obtained by the previously proposed training technique utilizing constant rhythmic auditory stimulation. Walk-Mate effectively compensated for locomotion problems of hemiparetic patients by improving gait rhythm both during and after training, suggesting that locomotive function can be effectively recovered in some patients. The interactive mechanism of Walk-Mate may be capable of simultaneously achieving the aims of gait compensation and gait rehabilitation training methods previously developed under individual frameworks. Walk-Mate is a promising technology for assisting the reintegration of disabled persons into society.


A High-Affinity Calmodulin-Binding Site in the CyaA Toxin Translocation Domain is Essential for Invasion of Eukaryotic Cells.
The molecular mechanisms and forces involved in the translocation of bacterial toxins into host cells are still a matter of intense research. The adenylate cyclase (CyaA) toxin from Bordetella pertussis displays a unique intoxication pathway in which its catalytic domain is directly translocated across target cell membranes. The CyaA translocation region contains a segment, P454 (residues 454-484), which exhibits membrane-active properties related to antimicrobial peptides. Herein, the results show that this peptide is able to translocate across membranes and to interact with calmodulin (CaM). Structural and biophysical analyses reveal the key residues of P454 involved in membrane destabilization and calmodulin binding. Mutational analysis demonstrates that these residues play a crucial role in CyaA translocation into target cells. In addition, calmidazolium, a calmodulin inhibitor, efficiently blocks CyaA internalization. It is proposed that after CyaA binding to target cells, the P454 segment destabilizes the plasma membrane, translocates across the lipid bilayer and binds calmodulin. Trapping of CyaA by the CaM:P454 interaction in the cytosol may assist the entry of the N-terminal catalytic domain by converting the stochastic motion of the polypeptide chain through the membrane into an efficient vectorial chain translocation into host cells.


Pre-operative renal function and selective renal vein renin levels as markers of favourable outcome in renovascular hypertension.
To evaluate factors that affect outcome following revascularization in patients with renovascular hypertension. This study was conducted at the Aga Khan University Hospital. It included all the patients diagnosed to have Renovascular hypertension confirmed by renal angiography between July 1996 to September 2000. Using ANOVA (Analysis of variance) and paired-sample t-test, several preoperative factors were evaluated for their ability to predict postoperative improvement of blood pressure and renal function. For descriptive purpose patients were divided into cured, improved and failure groups. Of the total 15 patients, 9 were males and 6 were females. Before the surgical procedure, 13 patients were on 3 or more antihypertensive drugs. Eleven patients received vein grafts, three patients had PTFE (polytetraflouroethyline) grafts while one patient underwent angioplasty and stenting. The patients had a median follow-up period of nine months with a range of 2 to 84 months. Five were completely cured, as they became normotensive without antihypertensive therapy after operation, whereas four showed marked improvement in BP control postoperatively, requiring two antihypertensive drugs. Six patients showed no improvement requiring 3 or more drugs. Extended cure or improvement of renovascular hypertension was achieved in nine patients. Normal preoperative serum creatinine level, high preoperative unstimulated peripheral renin levels and renal vein renin ratio of at least 1.75:1 were the most significant predictive factors for favorable outcome. This study confirms the long term effectiveness and safety of renovascular reconstruction in the relief of severe hypertension. The results further suggest that normal preoperative renal function, high renal vein renin ratio and high preoperative unstimulated peripheral renin levels are associated with the most favourable postoperative outcome (p <0.01). Preoperative severity and duration of hypertension and degree of disparity in kidney sizes do not affect the overall results in terms of postoperative improvement in renal function and blood pressure control (p=0.734).


Description of Gloeomargarita lithophora gen. nov., sp. nov., a thylakoid-bearing, basal-branching cyanobacterium with intracellular carbonates, and proposal for Gloeomargaritales ord. nov.
A unicellular cyanobacterium, strain Alchichica-D10, was isolated from microbialites of the alkaline Lake Alchichica, Mexico. The cells were short rods (3.9±0.6 µm in length and 1.1±0.1 µm in width) forming biofilms of intense emerald green colour. They exhibited red autofluorescence under UV light excitation. UV-visible absorption spectra revealed that they contain chlorophyll a and phycocyanin, and electron microscopy showed the presence of thylakoids. The strain grew within a temperature range of 15-30 °C. Genomic DNA G+C content was 52.2 mol%. The most remarkable feature of this species was its granular cytoplasm, due to the presence of numerous intracellular spherical granules (16-26 per cell) with an average diameter of 270 nm. These granules, easily visible under scanning electron microscopy, were composed of amorphous carbonate containing Ca, Mg, Ba and Sr. A multi-gene phylogeny based on the analysis of 59 conserved protein markers supported robustly that this strain occupies a deep position in the cyanobacterial tree. Based on its phenotypic characters and phylogenetic position, strain Alchichica-D10 is considered to represent a new genus and novel species of cyanobacteria for which the name Gloeomargarita lithophora gen. nov., sp. nov. is proposed. The type strain is Alchichica-D10 (Culture Collection of Algae and Protozoa CCAP strain 1437/1; Collections de Cyanobactéries et Microalgues Vivantes of the Museum National d'Histoire Naturelle in Paris strain PMC 919.15). Furthermore, a new family, Gloeomargaritaceae, and a new order, Gloeoemargaritales, are proposed to accommodate this species under the International Code of Nomenclature for algae, fungi and plants.


Evaluation of changes in vertical ground reaction forces as indicators of meniscal damage after transection of the cranial cruciate ligament in dogs.
To determine whether decreases in peak vertical force of the hind limb after transection of the cranial cruciate ligament (CrCL) would be indicative of medial meniscal damage in dogs. 39 purpose-bred adult male Walker Hounds. The right CrCL was transected arthroscopically. Force plate measurements of the right hind limb were made prior to and 2, 4, 10, and 18 weeks after transection of the CrCL. Only dogs with > or =10% decreases in peak vertical force after week 2 were considered to have potential meniscal damage. Dogs that did not have > or =10% decreases in peak vertical force at any time point after week 2 were assigned to group 1. Group 2 dogs had > or =10% decreases in peak vertical force from weeks 2 to 4 only. Group 3 and 4 dogs had > or =10% decreases in peak vertical force from weeks 4 to 10 only or from weeks 10 to 18 only, respectively. Damage to menisci and articular cartilage was graded at week 18, and grades for groups 2 to 4 were compared with those of group 1. The percentage change in peak vertical force and impulse area was significantly different in groups 2 (n = 4), 3 (4), and 4 (4) at the end of each measurement period (weeks 4, 10, and 18, respectively) than in group 1 (27). The meniscal grade for groups 2 to 4 was significantly higher than for group 1. A > or =10% decrease in peak vertical force had sensitivity of 52% and accuracy of 72% for identifying dogs with moderate to severe medial meniscal damage. In dogs with transected or ruptured CrCLs, force plate analysis can detect acute exacerbation of lameness, which may be the result of secondary meniscal damage, and provide an objective noninvasive technique that delineates the temporal pattern of medial meniscal injury.


Coordinate based random effect size meta-analysis of neuroimaging studies.
Low power in neuroimaging studies can make them difficult to interpret, and Coordinate based meta-analysis (CBMA) may go some way to mitigating this issue. CBMA has been used in many analyses to detect where published functional MRI or voxel-based morphometry studies testing similar hypotheses report significant summary results (coordinates) consistently. Only the reported coordinates and possibly t statistics are analysed, and statistical significance of clusters is determined by coordinate density. Here a method of performing coordinate based random effect size meta-analysis and meta-regression is introduced. The algorithm (ClusterZ) analyses both coordinates and reported t statistic or Z score, standardised by the number of subjects. Statistical significance is determined not by coordinate density, but by a random effects meta-analyses of reported effects performed cluster-wise using standard statistical methods and taking account of censoring inherent in the published summary results. Type 1 error control is achieved using the false cluster discovery rate (FCDR), which is based on the false discovery rate. This controls both the family wise error rate under the null hypothesis that coordinates are randomly drawn from a standard stereotaxic space, and the proportion of significant clusters that are expected under the null. Such control is necessary to avoid propagating and even amplifying the very issues motivating the meta-analysis in the first place. ClusterZ is demonstrated on both numerically simulated data and on real data from reports of grey matter loss in multiple sclerosis (MS) and syndromes suggestive of MS, and of painful stimulus in healthy controls. The software implementation is available to download and use freely.


An investigation into the use of low quantities of functional additives to control drug release from hot melt extruded solid dispersions for poorly soluble drug delivery.
The motivation of this study is to demonstrate the practicality of producing slow release and fast release products in a single-step hot melt extrusion (HME) process. HPMCAS as the carrier material showed good potential in monolithic controlled release formulations for the model drug, carbamazepine (CBZ). As binary formulations, CBZ-HPMCAS extrudates showed zero-order release over 24 h which was accompanied by the swelling of the extrudates. A range of functional excipients was used at low quantities to modulate the release rate. The release rates of the HME extrudates could be either accelerated by the incorporations of low quantities (5% w/w) of soluble additives or further sustained by adding lipid excipient, Gelucire 50/13. Clear phase separations of the soluble additives including crosscarmellose sodium, sodium starch glycolate, maltodextrin and lactose in the extrudates led to higher interior porosity and quicker erosion in comparison to the binary extrudates. The phase separated Gelucire in the extrudates led to the substantial swelling of the extrudates and resulted in further prolonged drug release. This study provided clear formulation strategies for modulating the drug release rate from controlled release formulation prepared directly by single-step HME. In addition, this research work also evaluates for the first time HME extrudates simultaneous swelling and drug release using this UV imaging technique. The whole dose cell of this instrumentation is utilised to provide insights into the dissolution process of solid dispersions prepared by HME.


Genital tract HIV RNA levels and their associations with human papillomavirus infection and risk of cervical precancer.
Plasma HIV RNA levels have been associated with the risk of human papillomavirus (HPV) and cervical neoplasia in HIV-seropositive women. However, little is known regarding local genital tract HIV RNA levels and their relation with cervical HPV and neoplasia. In an HIV-seropositive women's cohort with semiannual follow-up, we conducted a nested case-control study of genital tract HIV RNA levels and their relation with incident high-grade squamous intraepithelial lesions (HSIL) subclassified as severe (severe HSIL), as provided for under the Bethesda 2001 classification system. Specifically, 66 incidents of severe HSIL were matched to 130 controls by age, CD4 count, highly active antiretroviral therapy use, and other factors. We also studied HPV prevalence, incident detection, and persistence in a random sample of 250 subjects. Risk of severe HSIL was associated with genital tract HIV RNA levels (odds ratio comparing HIV RNA ≥ the median among women with detectable levels versus undetectable, 2.96; 95% confidence interval: 0.99 to 8.84; Ptrend = 0.03). However, this association became nonsignificant (Ptrend = 0.51) after adjustment for plasma HIV RNA levels. There was also no association between genital tract HIV RNA levels and the prevalence of any HPV or oncogenic HPV. However, the incident detection of any HPV (Ptrend = 0.02) and persistence of oncogenic HPV (Ptrend = 0.04) were associated with genital tract HIV RNA levels, after controlling plasma HIV RNA levels. These prospective data suggest that genital tract HIV RNA levels are not a significant independent risk factor for cervical precancer in HIV-seropositive women, but they leave open the possibility that they may modestly influence HPV infection, an early stage of cervical tumorigenesis.


Paeoniflorin suppresses inflammatory response in imiquimod-induced psoriasis-like mice and peripheral blood mononuclear cells (PBMCs) from psoriasis patients.
Psoriasis is one of the most common immune-mediated chronic inflammatory skin disorders, characterized by hyperproliferation of keratinocytes, dilation and growth of dermal capillary vasculature, and cellular infiltration of T cells, dendritic cells (DCs), and neutrophils. Paeoniflorin (PF), the principal component of total glucosides of paeony (TGP), displays anti-inflammatory and antioxidant properties in several animal models. In this study, we investigated the anti-inflammatory effects and mechanisms of PF in imiquimod (IMQ)-induced psoriasis-like mouse model. The effects of PF on inflammatory cytokine expression in peripheral blood mononuclear cells (PBMCs) from patients with psoriasis vulgaris were also observed. Our results indicated that PF effectively attenuated the clinical and histopathologic changes in IMQ-induced psoriasis-like mouse model. Furthermore, PF reduced the infiltration of T cells, CD11c(+)DCs, and neutrophils in lesional skin. In addition, PF also significantly decreased the mRNA expression of inflammatory cytokines, such as IL-17, INF-γ, IL-6, and TNF-α, in IMQ-induced psoriasis-like mouse model and PBMCs from patients with psoriasis vulgaris. Hence, our data suggest that PF can inhibit leukocyte infiltration and decrease the expression of inflammatory cytokines such as IL-17, INF-γ, IL-6, and TNF-α. PF might be a candidate drug for the treatment of psoriasis.


Habituation of reflexive and motivated behavior in mice with deficient BK channel function.
Habituation is considered the most basic form of learning. It describes the decrease of a behavioral response to a repeated non-threatening sensory stimulus and therefore provides an important sensory filtering mechanism. While some neuronal pathways mediating habituation are well described, underlying cellular/molecular mechanisms are not yet fully understood. In general, there is an agreement that short-term and long-term habituation are based on different mechanisms. Historically, a distinction has also been made between habituation of motivated versus reflexive behavior. In recent studies in invertebrates the large conductance voltage- and calcium-activated potassium (BK) channel has been implicated to be a key player in habituation by regulating synaptic transmission. Here, we tested mice deficient for the pore forming α-subunit of the BK channel for short-term and long-term habituation of the acoustic startle reflex (reflexive behavior) and of the exploratory locomotor behavior in the open field box (motivated behavior). Short-term habituation of startle was completely abolished in the BK knock-out mice, whereas neither long-term habituation of startle nor habituation of motivated behavior was affected by the BK deficiency. Our results support a highly preserved mechanism for short-term habituation of startle across species that is distinct from long-term habituation mechanisms. It also supports the notion that there are different mechanisms underlying habituation of motivated behavior versus reflexive behavior.


Elevated total and central adiposity and low physical activity are associated with insulin resistance in children.
The aim of this study was 2-fold: (1) to examine insulin resistance, blood lipid levels, and inflammatory markers in 9- to 11.5-year-old obese and lean children and (2) to identify factors that influence insulin resistance in this cohort of youths. Body mass index, skinfold thickness, waist circumference, physical activity (4-day triaxial accelerometer), cardiorespiratory fitness (submaximal bicycle ergometer test), and dietary intake (3-day food records) were evaluated in 27 obese and 27 lean boys and girls. Fasting blood samples were analyzed for insulin, glucose, lipids and lipoproteins, C-reactive protein (CRP), interleukin 6, soluble intercellular adhesion molecule, and soluble vascular cell adhesion molecule. Homeostasis model assessment (HOMA) was used to evaluate insulin resistance (HOMA-IR). Obese children presented higher HOMA-IR, CRP, and blood lipid levels (all P < .01) compared with lean children. Total body fat and waist circumference were positively associated with fasting insulin (r > or = 0.51), HOMA-IR (r > or = 0.56), CRP (r > or = 0.51), and blood triacylglycerol (r > or = 0.38), and were inversely correlated with high-density lipoprotein cholesterol (r > or = -0.39; all P < .01). Cardiorespiratory fitness was inversely associated with HOMA-IR (r = -0.24; P < .05), but this association disappeared when adjusted for age, sex, and fat mass. Waist circumference and total daily physical activity explained 49% of the variance in HOMA-IR in these children. In conclusion, these findings suggest that total and central adiposity are positively associated and physical activity is negatively associated with insulin resistance in children. Interventions to improve glucose metabolism in youth should target at reducing total body and abdominal fat and increasing physical activity. The lack of association between inflammatory markers and HOMA-IR suggests that obesity may precede the elevation of these markers in the evolution of insulin resistance in youth.


Rapid evolution of escape ability in Trinidadian guppies (Poecilia reticulata).
Predators are widely assumed to create selection that shapes the evolution of prey escape abilities. However, this assumption is difficult to test directly due to the challenge of recording both predation and its evolutionary consequences in the wild. We examined these events by studying natural and experimental populations of Trinidadian guppies, Poecilia reticulata, which occur in distinct high-predation and low-predation environments within streams. Importantly, in the last two decades several populations of guppies have been experimentally introduced from one type of predatory environment into the other, allowing measurements of the consequences of change. We used this system to test two hypotheses: First, that changes in predatory environments create phenotypic selection favoring changes in escape ability of guppies, and second, that this selection can result in rapid evolution. For the first test we compared escape ability of wild caught guppies from high- versus low-predation environments by measuring survival rates during staged encounters with a major predator, the pike cichlid Crenicichla alta. We used guppies from three streams, comparing two within-stream pairs of natural populations and three within-stream pairs of an introduced population versus its natural source population. In every comparison, guppies from the high-predation population showed higher survival. These multiple, parallel divergences in guppy survival phenotype suggest that predatory environment does create selection of escape ability. We tested our second hypothesis by rearing guppies in common garden conditions in the laboratory, then repeating the earlier experiments using the F2 generation. As before, each comparison resulted in higher survival of guppies descended from the high-predation populations, demonstrating that population differences in escape ability have a genetic basis. These results also show that escape ability can evolve very rapidly in nature, that is, within 26-36 generations in the introduced populations. Interestingly, we found rapid evolutionary loss of escape ability in populations introduced into low-predation environments, suggesting that steep fitness trade-offs may influence the evolution of escape traits.


Comparison of the Mallampati Classification in Sitting and Supine Position to Predict Difficult Tracheal Intubation: A Prospective Observational Cohort Study.
The Mallampati classification (MLPT) is normally evaluated in the sitting position. However, many patients cannot be evaluated in the sitting position for medical reasons. Thus, we compared the MLPT in sitting and supine positions in predicting difficult tracheal intubation (DTI). We hypothesized that the diagnostic accuracy of the MLPT performed in sitting and supine positions would differ. We performed a single-center prospective observational study in adult patients who received general anesthesia and orotracheal intubation for noncardiac surgery. During the preanesthesia consultation, the MLPT in the sitting position was recorded. The day of surgery, the MLPT in the supine position and the difficulty of intubation (DTI) were recorded by an independent observer. The diagnostic performance of the MLPT for the prediction of DTI was evaluated in the sitting and supine positions through the area under the receiver operating characteristic (ROC) curve. The performance of the Naguib score in predicting DTI was calculated with the MLPT in sitting and supine positions. Among the 3036 patients, 157 (5.1%) had DTI. The area under the ROC curve for the MLPT in supine position (0.82 [0.78-0.84]) was greater than that for the MLPT in the sitting position (0.70 [0.66-0.75]; P < .001). The relationship between the sitting and supine MLPTs was moderate (Spearman rank correlation coefficient: 0.50; P < .001). The area under ROC curve for predicting DTI by the Naguib score calculated with the supine MLPT (0.78 [95% confidence interval, 0.74-0.82]) was greater than that for the Naguib score calculated with MLPT in the sitting position (0.69 [95% confidence interval, 0.63-0.74)]; P < .001). The MLPT performed in the supine position is possibly superior to that performed in the sitting position for predicting difficult intubation in adults.


Influence of Polymer Molecular Weight on Drug-Polymer Solubility: A Comparison between Experimentally Determined Solubility in PVP and Prediction Derived from Solubility in Monomer.
In this study, the influence of polymer molecular weight on drug-polymer solubility was investigated using binary systems containing indomethacin (IMC) and polyvinylpyrrolidone (PVP) of different molecular weights. The experimental solubility in PVP, measured using a differential scanning calorimetry annealing method, was compared with the solubility calculated from the solubility of the drug in the liquid analogue N-vinylpyrrolidone (NVP). The experimental solubility of IMC in the low-molecular-weight PVP K12 was not significantly different from that in the higher molecular weight PVPs (K25, K30, and K90). The calculated solubilities derived from the solubility in NVP (0.31-0.32 g/g) were found to be lower than those experimentally determined in PVP (0.38-0.40 g/g). Nevertheless, the similarity between the values indicates that the analogue solubility can provide valuable indications on the solubility in the polymer. Hence, if a drug is soluble in an analogue of the polymer, it is most likely also soluble in the polymer. In conclusion, the solubility of a given drug-polymer system is determined by the strength of the drug-polymer interactions rather than the molecular weight of the polymer. Therefore, during the first screenings for drug solubility in polymers, only one representative molecular weight per polymer is needed.


Medical staff appointment and delineation of pediatric privileges in hospitals.
The review and verification of credentials and the granting of clinical privileges are required of every hospital to ensure that members of the medical staff are competent and qualified to provide specified levels of patient care. The credentialing process involves the following: (1) assessment of the professional and personal background of each practitioner seeking privileges; (2) assignment of privileges appropriate for the clinician's training and experience; (3) ongoing monitoring of the professional activities of each staff member; and (4) periodic reappointment to the medical staff on the basis of objectively measured performance. We examine the essential elements of a credentials review for initial and renewed medical staff appointments along with suggested criteria for the delineation of clinical privileges. Sample forms for the delineation of privileges can be found on the American Academy of Pediatrics Committee on Hospital Care Web site (http://www.aap.org/visit/cmte19.htm). Because of differences among individual hospitals, no 1 method for credentialing is universally applicable. The medical staff of each hospital must, therefore, establish its own process based on the general principles reviewed in this report. The issues of medical staff membership and credentialing have become very complex, and institutions and medical staffs are vulnerable to legal action. Consequently, it is advisable for hospitals and medical staffs to obtain expert legal advice when medical staff bylaws are constructed or revised.


Comparisons of self-reported and register data on sickness absence among public employees in Sweden.
Self-reported assessments of sickness absence are often performed in epidemiological studies. The objective of this study was to compare the number of sick-leave days according to self-reported data over 12 months with data from the employer's register for the same period. An additional aim was to ascertain whether the self-reported information and the recorded data would show equivalent associations with self-reported general health. The study was based on a cohort of 4869 municipal employees in Sweden, about 80% women, who answered a questionnaire in 2001-2. The responses provided by the employees included information on number of sick-leave days and self-rated health. Data on sick-leave days, occupation and age were derived from the employers' computerised registers. The questionnaire information on sick-leave days was compared with the corresponding information retrieved from the employer register by means of calculating sensitivity and specificity, using the employers' data as the "gold standard". The annual number of sick-leave days was lower according to the self-reported information than to the register data. For women the agreement between the two sickness absence measures for no sick-leave days, 1-7 days and >/=28 days were 74%, 72% and 67%, respectively. The sensitivity of questionnaire versus register information regarding any self-reported sick-leave day was 91% and the specificity was 74%. Sensitivity and specificity for sickness absence >/=28 days were 67% and 98%, respectively. The results for men were similar to those for women. Self-reported and recorded sickness absence were both associated with self-rated health. The odds ratios were 7.27 and 8.25, for subjects with >/=28 recorded and self-reported number of sick-leave days respectively, compared to subjects with no sickness absence. Good agreement was found between self-reported and register information on sickness absence. Self-reported data on sickness absence may be useful in common epidemiological applications.