Quantitation of hepatitis B virus DNA in plasma using a sensitive cost-effective "in-house" real-time PCR assay.
Sensitive nucleic acid testing for the detection and accurate quantitation of hepatitis B virus (HBV) is necessary to reduce transmission through blood and blood products and for monitoring patients on antiviral therapy. The aim of this study is to standardize an "in-house" real-time HBV polymerase chain reaction (PCR) for accurate quantitation and screening of HBV. The "in-house" real-time assay was compared with a commercial assay using 30 chronically infected individuals and 70 blood donors who are negative for hepatitis B surface antigen, hepatitis C virus (HCV) antibody and human immunodeficiency virus (HIV) antibody. Further, 30 HBV-genotyped samples were tested to evaluate the "in-house" assay's capacity to detect genotypes prevalent among individuals attending this tertiary care hospital. The lower limit of detection of this "in-house" HBV real-time PCR was assessed against the WHO international standard and found to be 50 IU/mL. The interassay and intra-assay coefficient of variation (CV) of this "in-house" assay ranged from 1.4% to 9.4% and 0.0% to 2.3%, respectively. Virus loads as estimated with this "in-house" HBV real-time assay correlated well with the commercial artus HBV RG PCR assay ( r = 0.95, P < 0.0001). This assay can be used for the detection and accurate quantitation of HBV viral loads in plasma samples. This assay can be employed for the screening of blood donations and can potentially be adapted to a multiplex format for simultaneous detection of HBV, HIV and HCV to reduce the cost of testing in blood banks.


Improved characterization of combat injury.
Combat injury patterns differ from civilian trauma in that the former are largely explosion-related, comprising multiple mechanistic and fragment injuries and high-kinetic-energy bullets. Further, unlike civilians, U.S. armed forces combatants are usually heavily protected with helmets and Kevlar body armor with ceramic plate inserts. Searchable databases providing actionable, statistically valid knowledge of body surface entry wounds and resulting organ injury severity are essential to understanding combat trauma. Two tools were developed to address these unique aspects of combat injury: (1) the Surface Wound Mapping (SWM) database and Surface Wound Analysis Tool (SWAT) software that were developed to generate 3D density maps of point-of-surface wound entry and resultant anatomic injury severity; and (2) the Abbreviated Injury Scale (AIS) 2005-Military that was developed by a panel of military trauma surgeons to account for multiple injury etiology from explosions and other high-kinetic- energy weapons. Combined data from the Joint Theater Trauma Registry, Navy/Marine Combat Trauma Registry, and the Armed Forces Medical Examiner System Mortality Trauma Registry were coded in AIS 2005-Military, entered into the SWM database, and analyzed for entrance site and wounding path. When data on 1,151 patients, who had a total of 3,500 surface wounds and 12,889 injuries, were entered into SWM, surface wounds averaged 3.0 per casualty and injuries averaged 11.2 per casualty. Of the 3,500 surface wounds, 2,496 (71%) were entrance wounds with 6,631 (51%) associated internal injuries, with 2.2 entrance wounds and 5.8 associated injuries per casualty (some details cannot be given because of operational security). Crude deaths rates were calculated using Maximum AIS-Military. These new tools have been successfully implemented to describe combat injury, mortality, and distribution of wounds and associated injuries. AIS 2005-Military is a more precise assignment of severity to military injuries. SWM has brought data from all three combat registries together into one analyzable database. SWM and SWAT allow visualization of wounds and associated injuries by region on a 3D model of the body.


Molecular and Physiological Analysis of the Powdery Mildew Antagonist Pseudozyma flocculosa and Related Fungi.
ABSTRACT A number of phenotypic and genotypic characteristics were used to ascertain the identity and diversity of Pseudozyma flocculosa, a natural antagonist of powdery mildews that has received little attention in terms of taxonomy. To this end, several putative isolates of P. flocculosa as well as several closely related species were analyzed. Ribosomal DNA sequences distinguished P. flocculosa from other Pseudozyma spp. and identified two previously unknown Pseudozyma isolates as P. flocculosa. Random amplified microsatellites revealed three distinct P. flocculosa strains among the tested isolates. Biocontrol properties and antifungal metabolite production were limited to the P. flocculosa spp. Results produced useful molecular markers to (i) distinguish P. flocculosa from other related fungi, (ii) identify different strains within this species, and (iii) aid in the construction of isolate-specific molecular tools that will assist in research and development of P. flocculosa as a biocontrol agent of powdery mildew fungi.


Applications of gold nanoparticles in cancer nanotechnology.
It has been almost 4 decades since the "war on cancer" was declared. It is now generally believed that personalized medicine is the future for cancer patient management. Possessing unprecedented potential for early detection, accurate diagnosis, and personalized treatment of cancer, nanoparticles have been extensively studied over the last decade. In this review, we will summarize the current state-of-the-art of gold nanoparticles in biomedical applications targeting cancer. Gold nanospheres, nanorods, nanoshells, nanocages, and surface enhanced Raman scattering nanoparticles will be discussed in detail regarding their uses in in vitro assays, ex vivo and in vivo imaging, cancer therapy, and drug delivery. Multifunctionality is the key feature of nanoparticle-based agents. Targeting ligands, imaging labels, therapeutic drugs, and other functionalities can all be integrated to allow for targeted molecular imaging and molecular therapy of cancer. Big strides have been made and many proof-of-principle studies have been successfully performed. The future looks brighter than ever yet many hurdles remain to be conquered. A multifunctional platform based on gold nanoparticles, with multiple receptor targeting, multimodality imaging, and multiple therapeutic entities, holds the promise for a "magic gold bullet" against cancer.


Epicardial development in the rat: a new perspective.
Development of the epicardium is critical to proper heart formation. It provides all of the precursor cells that form the coronary system and supplies signals that stimulate cardiac myocyte proliferation. The epicardium forms from mesothelial cells associated with the septum transversum and is referred to as the proepicardium (PE). Two different methods by which these PE cells colonize the developing heart have been described. In avians, PE cells form a bridge to the heart over which PE cells migrate onto the heart. In fish and mammals, PE cells form vesicles of cells that detach from the mesothelium, float through the pericardial cavity, and attach to the heart. A previous study of rat PE development investigated this process at the histological level. Protein markers have been developed since this study. Thus, we investigated this important developmental process coupled with these new markers using other visualization techniques such as scanning electron microscopy (SEM) and confocal microscopy. Finally, a novel, three-dimensional (3-D) culture system was used to confirm the identity of the PE cells. In this study, we found convincing evidence that the rat PE cells directly attach to the heart in a manner similar to that observed in avians.


Competitive binding of cations to duplex DNA revealed through molecular dynamics simulations.
The concept of "ion atmosphere" is prevalent in both theoretical and experimental studies of nucleic acid systems, yet the spatial arrangement and the composition of ions in the ion atmosphere remain elusive, in particular when several ionic species (e.g., Na(+), K(+), and Mg(2+)) compete to neutralize the charge of a nucleic acid polyanion. Complementing the experimental study of Bai and co-workers (J. Am. Chem. Soc.2007, 129, 14981), here we characterize ion atmosphere around double-stranded DNA through all-atom molecular dynamics simulations. We demonstrate that our improved parametrization of the all-atom model can quantitatively reproduce the experimental ion-count data. Our simulations determine the size of the ion atmosphere, the concentration profiles of ionic species competing to neutralize the DNA charge, and the sites of the cations' preferential binding at the surface of double-stranded DNA. We find that the effective size of the ion atmosphere depends on both the bulk concentration and valence of ions: increasing either reduces the size of the atmosphere. Near DNA, the concentration of Mg(2+) is strongly enhanced compared to monovalent cations. Within the DNA grooves, the relative concentrations of cations depend on their bulk values. Nevertheless, the total charge of competing cations buried in the DNA grooves is constant and compensates for about ~30% of the total DNA charge.


Durability of aneurysm embolization with matrix detachable coils.
Matrix detachable coils (MDC; Boston Scientific/Target, Fremont, CA) are platinum coils coated with a bioabsorbable polymeric material (polyglycolic-polylactic acid). In animal models, the introduction of polyglycolic-polylactic acid-coated coils into experimental aneurysms resulted in a cellular reaction which promoted stable intra-aneurysmal scar tissue formation. The current study was undertaken to assess the durability of aneurysm occlusion after embolization with MDC. All patients undergoing embolization were prospectively enrolled in an endovascular database at our institution. All coils used for embolization were recorded in the operative notes for the procedure. Only aneurysms embolized with 50% or greater length of MDC were included. All patients with Neuroform stents (Boston Scientific/Target, Fremont, CA) were excluded from the study. Patients were followed with conventional angiography and magnetic resonance angiography. During a 20-month period, 131 aneurysms were embolized with MDC. Follow up data (average, 6.9 mo; range, 1.5-22 mo) were available for 82 aneurysms (61 with conventional angiography, 21 with magnetic resonance angiography only). Of the aneurysm patients with follow-up available, 65 had small aneurysms with small necks, three were small aneurysms with wide necks, 12 were large aneurysms, and two were giant aneurysms. Overall, there were 30 (36.6%) recanalizations, 19 (23.1%) of which required retreatment. In two additional patients, retreatment was either recommended (n = 1) or attempted unsuccessfully (n = 1). Fifty-two aneurysms were unchanged or demonstrated progressive thrombosis (63.4%). The recanalization rate for small aneurysms with small necks was 26.1% (17 out of 65) with a 13.8% retreatment rate (9/65). The recanalization rate for large aneurysms was 75% (9 out of 12) with seven requiring retreatment. In the absence of Neuroform stent support, aneurysms embolized with the MDC system demonstrated a significant rate of recanalization. Many of the recanalizations were of sufficient size to warrant retreatment. The rates of recanalization observed in the present series were comparable to, or worse than, those reported for bare platinum coils.


Different GPI-attachment signals affect the oligomerisation of GPI-anchored proteins and their apical sorting.
To understand the mechanism involved in the apical sorting of glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) we fused to the C-terminus of GFP the GPI-anchor-attachment signal of the folate receptor (FR) or of the prion protein (PrP), two native GPI-anchored proteins that are sorted apically or basolaterally, respectively, in MDCK cells. We investigated the behaviour of the resulting fusion proteins GFP-FR and GFP-PrP by analysing three parameters: their association with DRMs, their oligomerisation and their apical sorting. Strikingly, we found that different GPI-attachment signals differently modulate the ability of the resulting GFP-fusion protein to oligomerise and to be apically sorted. This is probably owing to differences in the GPI anchor and/or in the surrounding lipid microenvironment. Accordingly, we show that addition of cholesterol to the cells is necessary and sufficient to drive the oligomerisation and consequent apical sorting of GFP-PrP, which under control conditions does not oligomerise and is basolaterally sorted.


Locus-specific and activity-independent gene repositioning during early tumorigenesis.
The mammalian genome is highly organized within the cell nucleus. The nuclear position of many genes and genomic regions changes during physiological processes such as proliferation, differentiation, and disease. It is unclear whether disease-associated positioning changes occur specifically or are part of more global genome reorganization events. Here, we have analyzed the spatial position of a defined set of cancer-associated genes in an established mammary epithelial three-dimensional cell culture model of the early stages of breast cancer. We find that the genome is globally reorganized during normal and tumorigenic epithelial differentiation. Systematic mapping of changes in spatial positioning of cancer-associated genes reveals gene-specific positioning behavior and we identify several genes that are specifically repositioned during tumorigenesis. Alterations of spatial positioning patterns during differentiation and tumorigenesis were unrelated to gene activity. Our results demonstrate the existence of activity-independent genome repositioning events in the early stages of tumor formation.


A new hybrid mouse model for agenesis of the corpus callosum.
A three locus model of the inheritance of absent corpus callosum in mice was tested by creating F1 and F2 hybrid crosses from the strains BALB/cWah1 and 129/J which show incomplete penetrance for callosal agenesis. The model predicted that a few of the F2 hybrid mice would suffer severe reduction of the hippocampal commissure when the corpus callosum was absent, a condition that usually occurs only in the most consistently acallosal I/LnJ strain, and this prediction was confirmed. The C129F2 hybrid population expresses substantial genetic variation and an extremely wide range of defects of the corpus callosum, dorsal commissure of the fornix and hippocampal commissure. At the same time, these hybrids have exceptionally good health and reproductive performance, unlike their inbred parent strains. These characteristics make them ideal subjects for the study of brain-behaviour correlation using a noninvasive method.


Surgeon perceptions of Natural Orifice Translumenal Endoscopic Surgery (NOTES).
If proven feasible and safe, Natural Orifice Translumenal Endoscopic Surgery (NOTES) would still need acceptance by surgeons if it were to become a mainstream approach. Three hundred fifty-seven surgeons responded to a preliminary survey describing NOTES and were asked to rate the importance of various surgical considerations and (assuming availability and safety) if they would choose to undergo and/or perform cholecystectomies by NOTES or laparoscopy and why. The risk of having a complication was considered most important. NOTES was theorized to be riskier and to require greater skill than laparoscopy but to potentially cause less pain and convalescence. Nearly three-fourths (72%) of surgeons expressed interest in NOTES training which correlated with younger age, SAGES membership, minimally invasive surgery specialization, and flexible endoscopic volume. Forty-four percent would like to introduce NOTES cholecystectomy into their practices. Among those not preferring NOTES, 88% would adopt NOTES if data showed improved outcomes over laparoscopy. Finally, only 24% would choose to undergo cholecystectomy themselves by NOTES, believing it to be too new and riskier than laparoscopy. The risk of having a complication is the greatest concern among surgeons, and safety will affect NOTES acceptance. The results of this survey seem to justify more focused future investigations.


Early initiation of prophylactic anticoagulation for prevention of coronavirus disease 2019 mortality in patients admitted to hospital in the United States: cohort study.
To evaluate whether early initiation of prophylactic anticoagulation compared with no anticoagulation was associated with decreased risk of death among patients admitted to hospital with coronavirus disease 2019 (covid-19) in the United States. Observational cohort study. Nationwide cohort of patients receiving care in the Department of Veterans Affairs, a large integrated national healthcare system. All 4297 patients admitted to hospital from 1 March to 31 July 2020 with laboratory confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and without a history of anticoagulation. The main outcome was 30 day mortality. Secondary outcomes were inpatient mortality, initiating therapeutic anticoagulation (a proxy for clinical deterioration, including thromboembolic events), and bleeding that required transfusion. Of 4297 patients admitted to hospital with covid-19, 3627 (84.4%) received prophylactic anticoagulation within 24 hours of admission. More than 99% (n=3600) of treated patients received subcutaneous heparin or enoxaparin. 622 deaths occurred within 30 days of hospital admission, 513 among those who received prophylactic anticoagulation. Most deaths (510/622, 82%) occurred during hospital stay. Using inverse probability of treatment weighted analyses, the cumulative incidence of mortality at 30 days was 14.3% (95% confidence interval 13.1% to 15.5%) among those who received prophylactic anticoagulation and 18.7% (15.1% to 22.9%) among those who did not. Compared with patients who did not receive prophylactic anticoagulation, those who did had a 27% decreased risk for 30 day mortality (hazard ratio 0.73, 95% confidence interval 0.66 to 0.81). Similar associations were found for inpatient mortality and initiation of therapeutic anticoagulation. Receipt of prophylactic anticoagulation was not associated with increased risk of bleeding that required transfusion (hazard ratio 0.87, 0.71 to 1.05). Quantitative bias analysis showed that results were robust to unmeasured confounding (e-value lower 95% confidence interval 1.77 for 30 day mortality). Results persisted in several sensitivity analyses. Early initiation of prophylactic anticoagulation compared with no anticoagulation among patients admitted to hospital with covid-19 was associated with a decreased risk of 30 day mortality and no increased risk of serious bleeding events. These findings provide strong real world evidence to support guidelines recommending the use of prophylactic anticoagulation as initial treatment for patients with covid-19 on hospital admission.


Hepatitis C virus prevalence and estimated incidence among new injectors during the opioid epidemic in New York City, 2000-2017: Protective effects of non-injecting drug use.
Assess hepatitis C virus (HCV) prevalence and incidence among person who began injecting drugs during the opioid epidemic in New York City (NYC) and identify possible new directions for reducing HCV infection among persons who inject drugs. 846 persons who began injecting drugs between 2000 and 2017 were recruited from persons entering Mount Sinai Beth Israel substance use treatment programs. A structured interview was administered and HCV antibody testing conducted. Protective effects of non-injecting drug use were examined among persons who "reversed transitioned" to non-injecting drug use and persons who used non-injected heroin in addition to injecting. Participants were 79% male, 41% White, 15% African-American, 40% Latinx, with a mean age of 35. Of those who began injecting in 2000 or later, 97 persons (11%) "reverse transitioned" back to non-injecting drug use. Reverse transitioning was strongly associated with lower HCV seroprevalence (30% versus 47% among those who continued injecting, p < 0.005). Among those who continued injecting, HCV seropositivity was inversely associated with current non-injecting heroin use (AOR = 0.72, 95%CI 0.52-0.99). HCV incidence among persons continuing to inject was estimated as 13/100 person-years. HCV seropositive persons currently injecting cocaine were particularly likely to report behavior likely to transmit HCV. Similar to other locations in the US, NYC is experiencing high rates of HCV infection among persons who have begun injecting since 2000. New interventions that facilitate substitution of non-injecting for injecting drug use and that reduce transmission behavior among HCV seropositives may provide additional methods for reducing HCV transmission.


Middle-atmospheric Doppler lidar using an iodine-vapor edge filter.
We present both modeled capabilities of and experimental data from a Doppler lidar for the stratosphere and the lower mesosphere that uses the edge of a molecular iodine filter in a differential measurement to provide frequency discrimination. Modeled results show a capability for wind measurements to an altitude of 55 km with 1.5-km resolution in 30 min. Experimentally, wind-vector components from 18 to 45 km are measured every 20 min. The molecular-vapor filter provides great advantages with regard to system stability, operation in less-than-optimum weather conditions, and simplicity of data analysis.


The prevalence of stunting is high in HIV-1-exposed uninfected infants in Kenya.
As prevention of mother-to-child HIV-1 transmission (PMTCT) programs decrease the numbers of HIV-1-infected infants, it remains important to improve growth in HIV-1-exposed, uninfected (EU) infants. To determine the growth rate and predictors of growth faltering in breast-fed and formula-fed EU infants, growth analyses [weight-for-age (WAZ), weight-for-length (WLZ), and length-for-age (LAZ) Z-scores] were conducted by using data from a randomized feeding trial in HIV-1-infected women in Kenya. Growth faltering in EU infants was compared based on randomization to breastfeeding (BF) or formula feeding (FF) using Cox proportional hazards regression models. Linear mixed-effects models determined rate and cofactors of length growth. Among 338 EU infants, 164 (49%) were breast-fed and 174 (51%) formula-fed. In both arms, growth declined steadily during follow-up. By 2 y, 29% of children were underweight (WAZ < -2), 18% were wasted (WLZ < -2), and 58% were stunted (LAZ < -2), with no differences by feeding arm. Higher maternal education (y) and taller stature (cm) were associated with a decreased risk of underweight and stunting [underweight: adjusted HR (aHR) = 0.90 (95% CI: 0.83, 0.99), P = 0.03, and aHR = 0.92 (95% CI: 0.87, 0.97), P = 0.002; and stunting: aHR = 0.91 (95% CI: 0.85, 0.97), P = 0.003, and aHR = 0.96 (95% CI: 0.92, 0.99), P = 0.02, respectively]. Diarrhea was associated with an increased risk of wasting [aHR = 2.26 (95% CI: 1.11, 4.62), P = 0.03]. In multivariate analyses, FF was associated with slower declines in length velocity [0.24 LAZ/y (95% CI: 0.06, 0.43), P = 0.009]. Despite being uninfected, HIV-1-exposed infants showed frequent growth faltering, suggesting the need for vigilance in recognizing stunting within PMTCT programs. The slower rate of decline in length growth with FF may reflect benefits of micronutrients. Because BF is the best option for HIV-1-infected mothers in resource-limited settings, nutritional interventions should be examined for their impact on growth in EU breast-fed infants.


Immunoglobulin A-induced shift of Epstein-Barr virus tissue tropism.
Increased immunoglobulin A (IgA) antibodies to the Epstein-Barr virus (EBV) appear months to years before the clinical onset of nasopharyngeal carcinoma and define populations at high risk for this EBV-associated epithelial cancer common in south China. In the human HT-29 epithelial cell line, polymeric IgA (pIgA) specific for EBV promoted infection of the otherwise refractory epithelial cells. When bound to pIgA, EBV entered epithelial cells through secretory component-mediated IgA transport but no longer infected B lymphocytes. Such an immune-induced shift in EBV tissue tropism provides a paradigm for endogenous spread of EBV in the immune host that predicts infectious sequelae of epithelium.


A tale of two counties: expanding health insurance coverage for children in California.
During difficult economic times, many California counties have expanded health insurance coverage for low-income children. These Children's Health Initiatives (CHIs) enroll children in public programs and provide new health insurance, Healthy Kids, for those ineligible for existing programs. This article describes the policy issues in implementing the Santa Clara and San Mateo County CHIs, as well as the children's enrollment levels and utilization of services. These CHIs are among the first of the thirty California counties planning or implementing such initiatives. Their success depends on leadership from county agencies that have not traditionally worked closely together, as well as the development of a diverse public and private funding base. This effort to provide universal coverage for all children is important to national policymakers desiring similar goals.


Analysis of sperm motility using optical tweezers.
This study examines the use of optical trapping as a quantitative measure of sperm motility. The effects of laser trap duration and laser trapping power on sperm motility are described between sperm swimming force, swimmimg speed, and speed of progression (SOP) score. Sperm (SOP scores of 2-4) were trapped by a continuous-wave 1064 nm single-point gradient laser trap. Trap duration effects were quantified for 15, 10, and 5 seconds at 420 mW laser power. Laser power effects were quantified at powers of 420 mW, 350 mW, 300 mW, and 250 mW for five seconds. Swimming force, swimming speed, and SOP score relationships were examined at a trap duration and trapping power shown to minimally affect sperm motility. Swimming forces were measured by trapping sperm and subsequently decreasing laser power until the sperm escaped the trap. Swimming trajectories were calculated by custom-built software, and SOP scores were assigned by three qualified sperm scoring experts. A ubiquitous class of sperm were identified that swim with relatively high forces that are uncorrelated to swimming speed. It is concluded that sperm swimming forces measured by optical trapping provide new and valuable quantitative information to assess sperm motility.


Genetic influences on atrophy patterns in familial Alzheimer's disease: a comparison of APP and PSEN1 mutations.
Mutations in the presenilin1 (PSEN1) and amyloid β-protein precursor (APP) genes account for the majority of cases of autosomal dominantly inherited Alzheimer's disease (AD). We wished to assess and compare the patterns of cerebral loss produced by these two groups of mutations. Volumetric magnetic resonance imaging and neuropsychological assessments were performed in individuals with clinical AD carrying mutations in the APP (n = 10) and PSEN1 (n = 18) genes and in healthy controls (n = 18). Voxel-based morphometry (VBM), cortical thickness, and region of interest analyses were performed. Mini-Mental State Examination scores were similar in the two disease groups suggesting similar levels of disease severity. There was evidence that APP subjects have smaller hippocampal volume compared with PSEN1 subjects (p = 0.007), and weak evidence that they have larger whole-brain and grey matter volumes (both p = 0.07). Although there was no evidence of statistically significant differences between APP and PSEN1 in VBM or cortical thickness analyses, effect-maps were suggestive of APP subjects having more medial temporal lobe atrophy and conversely PSEN1 subjects showing more neocortical loss. Neuropsychological data were consistent with these regional differences and suggested greater memory deficits in the APP patients and greater impairment in non-memory domains in the PSEN1 group, although these differences were not statistically significant. We conclude that the mechanisms by which APP and PSEN1 mutations cause neuronal loss may differ which furthers our understanding of the neuropathology underlying AD and may inform future therapeutic strategies and trial designs.


Prospective estimation of rates of change in mammographic parenchymal patterns: influence of age and of hormone replacement therapy.
The objective of this study was to assess the effect of age, breast size and use of hormone replacement therapy (HRT) on the rate of change of mammographic parenchymal patterns, and the effect of age on the probability of misclassification between patterns. It was designed as a longitudinal study of the members of the treatment arm of a non-randomized screening trial in which subjects were assigned to screening or not by year of birth. The subjects were women in the Kotka district of Finland, each of whom attended for four or five mammographic screens. Participants were all women living in the district who were born in the relevant years and accepted an invitation to screening. A model was fitted to the longitudinal data comprising the observed Wolfe patterns on each woman, with age and breast size as predictors of breast density at first screen, age and HRT use as predictors of change in density at future screens, and age as a predictor of misclassification of true density between favourable (non-dense) and unfavourable (dense) patterns (according to the Wolfe classification). Relevant posterior probability estimates (with 95% credible intervals) were as follows. The probability that a woman of age 43.5 is truly in the favourable state ranged from 0.35 (0.34-0.37) for smallest breast size to 0.74 (0.72-0.76) for the largest. The probability that a woman is truly in the favourable state at first screen increased from 0.37 (0.36-0.38) at age 40 to 0.59 (0.58-0.60) at age 47. The probability that a woman having a later screen who had truly been in the unfavourable state at her previous screen changed to the favourable state increased from 0.12 (0.11-0.13) at age 42 to 0.48 (0.46-0.50) at age 55 for a woman not taking HRT, and from 0.10 (0.09-0.11) to 0.43 (0.40-0.45) at the same ages for a woman taking HRT. The probability that a woman would have changed from being truly in the favourable state to the unfavourable state was 0.003 (0.001-0.003) for any age and HRT use. The probability that a woman truly in a favourable state would be correctly classified rose from 0.87 (0.85-0.89) at age 40 to 0.998 (0.997-0.998) at age 55. The probability that a woman truly in the unfavourable state would be correctly classified decreased from 0.96 (0.95-0.97) to 0.93 (0.91-0.94) between the same ages. The probability of being in a non-dense, favourable state increases with age, as does the rate of change from dense to non-dense patterns. These are consistent with previous work. The probability of non-dense patterns and the rate of change to non-dense patterns are reduced with HRT use. Errors of classification are relatively rare, but are dependent on the age of the subject.


Structural determinants of the eosinophil cationic protein antimicrobial activity.
Antimicrobial RNases are small cationic proteins belonging to the vertebrate RNase A superfamily and endowed with a wide range of antipathogen activities. Vertebrate RNases, while sharing the active site architecture, are found to display a variety of noncatalytical biological properties, providing an excellent example of multitask proteins. The antibacterial activity of distant related RNases suggested that the family evolved from an ancestral host-defence function. The review provides a structural insight into antimicrobial RNases, taking as a reference the human RNase 3, also named eosinophil cationic protein (ECP). A particular high binding affinity against bacterial wall structures mediates the protein action. In particular, the interaction with the lipopolysaccharides at the Gram-negative outer membrane correlates with the protein antimicrobial and specific cell agglutinating activity. Although a direct mechanical action at the bacteria wall seems to be sufficient to trigger bacterial death, a potential intracellular target cannot be discarded. Indeed, the cationic clusters at the protein surface may serve both to interact with nucleic acids and cell surface heterosaccharides. Sequence determinants for ECP activity were screened by prediction tools, proteolysis and peptide synthesis. Docking results are complementing the structural analysis to delineate the protein anchoring sites for anionic targets of biological significance.


Estimation of background serum 2,3,7,8-TCDD concentrations by using quantile regression in the UMDES and NHANES populations.
The goal of the present study was to quantify the population-based background serum concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by using data from the reference population of the 2005 University of Michigan Dioxin Exposure Study (UMDES) and the 2003-2004 National Health and Nutrition Examination Survey (NHANES). Multiple imputation was used to impute the serum TCDD concentrations below the limit of detection by combining the 2 data sources. The background mean, quartiles, and 95th percentile serum TCDD concentrations were estimated by age and sex by using linear and quantile regressions for complex survey data. Any age- and sex-specific mean, quartiles, and 95th percentiles of background serum TCDD concentrations of study participants between ages 18 and 85 years can be estimated from the regressions for the UMDES reference population and the NHANES non-Hispanic white population. For example, for a 50-year-old man in the reference population of UMDES, the mean, quartiles, and 95th percentile serum TCDD concentrations are estimated to be 1.1, 0.6, 1.1, 1.8, and 3.3 parts per trillion, respectively. The study also shows that the UMDES reference population is a valid reference population for serum TCDD concentrations for other predominantly white populations in Michigan. The serum TCDD concentrations increased with age and increased more over age in women than in men, and hence estimation of background concentrations must be adjusted for age and sex. The methods and results discussed in this article have wide application in studies of the concentrations of chemicals in human serum and in environmental samples.


Point-of-care cardiac ultrasound techniques in the physical examination: better at the bedside.
The development of hand-carried, battery-powered ultrasound devices has created a new practice in ultrasound diagnostic imaging, called 'point-of-care' ultrasound (POCUS). Capitalising on device portability, POCUS is marked by brief and limited ultrasound imaging performed by the physician at the bedside to increase diagnostic accuracy and expediency. The natural evolution of POCUS techniques in general medicine, particularly with pocket-sized devices, may be in the development of a basic ultrasound examination similar to the use of the binaural stethoscope. This paper will specifically review how POCUS improves the limited sensitivity of the current practice of traditional cardiac physical examination by both cardiologists and non-cardiologists. Signs of left ventricular systolic dysfunction, left atrial enlargement, lung congestion and elevated central venous pressures are often missed by physical techniques but can be easily detected by POCUS and have prognostic and treatment implications. Creating a general set of repetitive imaging skills for these entities for application on all patients during routine examination will standardise and reduce heterogeneity in cardiac bedside ultrasound applications, simplify teaching curricula, enhance learning and recollection, and unify competency thresholds and practice. The addition of POCUS to standard physical examination techniques in cardiovascular medicine will result in an ultrasound-augmented cardiac physical examination that reaffirms the value of bedside diagnosis.


Complete clearance and Psoriasis Area and Severity Index response for brodalumab and ustekinumab by previous treatment history in AMAGINE-2 and AMAGINE-3.
The pathway for treatment of psoriasis is partly dependent upon disease severity, and patients may experience inadequate response at any point along the treatment pathway. Patients who repeatedly fail therapy represent a population in whom effective and well-tolerated treatment options are limited. To investigate and describe patients achieving Psoriasis Area and Severity Index (PASI) 100 and cumulative treatment benefit over time in patients with moderate-to-severe psoriasis receiving brodalumab or ustekinumab by prior treatment. We conducted a post hoc analysis of data from two phase 3, randomized, controlled, 52-week AMAGINE trials of brodalumab to describe patients who achieved complete clearance as measured by PASI 100 by prior treatment subgroup (naïve to systemic and biologic treatment, systemic-treated but biologic-naïve, biologic-treated without failure, and biologic-treated with failure). A competing risk model was used to assess cumulative incidence over a 52-week period with outcomes of PASI 100 or inadequate response. Cumulative clinical benefit of treatment was determined with an area under the curve analysis. The 52-week cumulative incidence of patients achieving PASI 100 was consistently higher for brodalumab vs. ustekinumab across treatment pathway subgroups (76% vs. 58% in systemic/biologic-naïve patients, 78% vs. 55% in systemic-treated/biologic-naïve patients, 75% vs. 41% in biologic-treated patients without failure, and 70% vs. 30% in biologic-treated patients with failure). Rates of inadequate response were lower with brodalumab compared with ustekinumab across all subgroups. Cumulative treatment benefit was also higher for all subgroups treated with brodalumab compared with those treated with ustekinumab. Treatment with brodalumab was associated with higher levels of complete clearance and greater cumulative benefit over time compared with ustekinumab, in patients with moderate-to-severe psoriasis, regardless of prior treatment experience.


[Curative effect of alpha-lipoic acid on peripheral neuropathy in type 2 diabetes: a clinical study].
To evaluate the effect of alpha-lipoic acid (ALA) on diabetic peripheral neuropathy (DPN). 95 type 2 diabetic patients complicated with DPN were randomly divided into 2 groups: treatment group (n = 52), receiving ALA 600 mg/d in normal saline 250 ml given by intravenous drip infusion once a day for 14 days; and control group (n = 43), administered with radix salviae 20 ml by intravenous drip infusion once a day for 14 days. Before and 7 and 14 days after the management fasting glucose (FPG), fasting insulin (FINS), supersensitive C reactive protein (sCRP), and HbA1c were measured, and total symptom score (TSS) and Michigan neuropathy screening instrument (MNSI) were used to evaluate the nervous symptoms. 50 cases of the treatment group and 43 control cases completed the study. Compared with the control group, the TSS of the treatment group reduced significantly after 1 week, but the MNSI score did not change significantly. The TSS 7d after ALA administration of the treatment group was significantly lower than that before the ALA treatment in the same group and that of the control group 7 days after the radix salviae infusion as well (both P < 0.05). The TSS and MNSI score 14 days after the management of the treatment group were significantly lower than those of the control group (both P < 0.01). The symptom scores of numbness, sting sensation, and burning sensation reduced significantly 2-weeks after ALA treatment, but there were no significant differences in these symptoms after the management in the control group. The total gratification rate of the treatment group was 90% (45/50), significantly higher than hat of the control group (13.95%, 6/43, P < 0.01). One patient of the 50 patients receiving ALA treatment felt chest distress 2 days after ALA administration, but the symptom was improved in the same day after the velocity of intravenous drip was slowed down, and no other adverse effects were found in these two groups. ALA effectively improves the sensitive symptoms of DPN patients and is safe for most of the patients.


Significant accumulation of C(4)-specific pyruvate, orthophosphate dikinase in a C(3) plant, rice.
The C(4)-Pdk gene encoding the C(4) enzyme pyruvate, orthophosphate dikinase (PPDK) of maize (Zea mays cv Golden Cross Bantam) was introduced into the C(3) plant, rice (Oryza sativa cv Kitaake). When the intact maize C(4)-Pdk gene, containing its own promoter and terminator sequences and exon/intron structure, was introduced, the PPDK activity in the leaves of some transgenic lines was greatly increased, in one line reaching 40-fold over that of wild-type plants. In a homozygous line, the PPDK protein accounted for 35% of total leaf-soluble protein or 16% of total leaf nitrogen. In contrast, introduction of a chimeric gene containing the full-length cDNA of the maize PPDK fused to the maize C(4)-Pdk promoter or the rice Cab promoter only increased PPDK activity and protein level slightly. These observations suggest that the intron(s) or the terminator sequence of the maize gene, or a combination of both, is necessary for high-level expression. In maize and transgenic rice plants carrying the intact maize gene, the level of transcript in the leaves per copy of the maize C(4)-Pdk gene was comparable, and the maize gene was expressed in a similar organ-specific manner. These results suggest that the maize C(4)-Pdk gene behaves in a quantitatively and qualitatively similar way in maize and transgenic rice plants. The activity of the maize PPDK protein expressed in rice leaves was light/dark regulated as it is in maize. This is the first reported evidence for the presence of an endogenous PPDK regulatory protein in a C(3) plant.


Cytosolic p120-catenin regulates growth of metastatic lobular carcinoma through Rock1-mediated anoikis resistance.
Metastatic breast cancer is the major cause of cancer-related death among women in the Western world. Invasive carcinoma cells are able to counteract apoptotic signals in the absence of anchorage, enabling cell survival during invasion and dissemination. Although loss of E-cadherin is a cardinal event in the development and progression of invasive lobular carcinoma (ILC), little is known about the underlying mechanisms that govern these processes. Using a mouse model of human ILC, we show here that cytosolic p120-catenin (p120) regulates tumor growth upon loss of E-cadherin through the induction of anoikis resistance. p120 conferred anchorage independence by indirect activation of Rho/Rock signaling through interaction and inhibition of myosin phosphatase Rho-interacting protein (Mrip), an antagonist of Rho/Rock function. Consistent with these data, primary human ILC samples expressed hallmarks of active Rock signaling, and Rock controlled the anoikis resistance of human ILC cells. Thus, we have linked loss of E-cadherin - an initiating event in ILC development - to Rho/Rock-mediated control of anchorage-independent survival. Because activation of Rho and Rock are strongly linked to cancer progression and are susceptible to pharmacological inhibition, these insights may have clinical implications for the development of tailor-made intervention strategies to better treat invasive and metastatic lobular breast cancer.


Protocol: a realist review of user fee exemption policies for health services in Africa.
Background Four years prior to the Millenium Development Goals (MDGs) deadline, low- and middle-income countries and international stakeholders are looking for evidence-based policies to improve access to healthcare for the most vulnerable populations. User fee exemption policies are one of the potential solutions. However, the evidence is disparate, and systematic reviews have failed to provide valuable lessons. The authors propose to produce an innovative synthesis of the available evidence on user fee exemption policies in Africa to feed the policy-making process. Methods The authors will carry out a realist review to answer the following research question: what are the outcomes of user fee exemption policies implemented in Africa? why do they produce such outcomes? and what contextual elements come into play? This type of review aims to understand how contextual elements influence the production of outcomes through the activation of specific mechanisms, in the form of context-mechanism-outcome configurations. The review will be conducted in five steps: (1) identifying with key stakeholders the mechanisms underlying user fee exemption policies to develop the analytical framework, (2) searching for and selecting primary data, (3) assessing the quality of evidence using the Mixed-Method Appraisal Tool, (4) extracting the data using the analytical framework and (5) synthesising the data in the form of context-mechanism-outcomes configurations. The output will be a middle-range theory specifying how user fee exemption policies work, for what populations and under what circumstances. Ethics and dissemination The two main target audiences are researchers who are looking for examples to implement a realist review, and policy-makers and international stakeholders looking for lessons learnt on user fee exemption. For the latter, a knowledge-sharing strategy involving local scientific and policy networks will be implemented. The study has been approved by the ethics committee of the CHUM Research Centre (CR-CHUM). It received funding from the Canadian Institutes of Health Research. The funders will not have any role in study design; collection, management, analysis, and interpretation of data; writing of the report and the decision to submit the report for publication, including who will have ultimate authority over each of these activities.


Demonstration of the in vitro phagocytosis of Treponema pallidum by rabbit peritoneal macrophages.
Evidence has been provided for the in vitro phagocytosis of virulent Treponema pallidum by stimulant-induced peritoneal macrophages. After the 4-hr incubation of macrophages with T. pallidum, treponemal antigens associated with the macrophages are specifically stained using indirect immunofluorescent techniques. Phagocytized treponemes appear within the cytoplasm of macrophages as round, brightly fluorescent "bodies" observable in increasing numbers as the duration of the treponeme-phagocyte interaction increases. Their presence is significantly reduced in the cytoplasm of macrophages that have been treated with cytochalasin B, a known inhibitor of phagocytosis, and in nonphagocytic fibroblasts. Additionally, supportive evidence for T. pallidum phagocytosis in vitro has been provided by electron microscopic examination in which treponemes have been demonstrated within typical phagocytic vacuoles. This study also provides evidence that immune serum factor(s) significantly promote the phagocytosis of T. pallidum, although a contribution by heat-labile serum factors has not been demonstrated. The possible mechanisms of immune serum contribution and the implications of the demonstration of T. pallidum phagocytosis are discussed.


Short rib-polydactyly syndrome: a case report.
Short rib-polydactyly syndrome (SRPS) is a group of rare, lethal skeletal dysplasias characterized by short ribs and limbs, polydactyly, hypoplastic thorax and visceral anomalies. Our case had coarsening of facial features, low-set ears, lobulated tongue, cleft palate, and hypoplastic epiglottis. Short proximal parts of upper limbs, bilateral postaxial polydactyly of hands, and bifid big toe with zygodactyly were additional findings. Chest was narrow. Ambiguous genitalia was noted but testicles were in scrotum. Choroid plexus cyst and coarctation of aorta were found in autopsy. Radiographies of the skull revealed occipital horn accompanied by prominent external occipital protuberance. The thoracic cage was narrow and elongated with short and iliac wings, pubic and ischial rami were were hypoplastic, and both acetabula were shallow and trident shaped. All tubular bones had wide and rounded metaphyses. Because clinical and radiological features of the four established subtypes are very similar, there are difficulties in the classification. We report an infant whose radiological, clinical and postmortem features were consistent with type IV SRPS (Beemer-Langer).


Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts.
Several proteases, i.e., pronase, a mixture of trypsin and chymotrypsin, and thermolysin were screened as potential surface probes of isolated intact pea (Pisum sativum var Laxton's Progress No. 9) chloroplasts. Of these, only thermolysin met the criteria of a suitable probe. Thermolysin destroyed outer envelope polypeptides, but did not affect inner envelope polypeptides, envelope permeability properties or such chloroplast activities as metabolite transport and O(2) evolution.


The relationship between muscarinic receptor occupancy and adenylate cyclase inhibition in the rabbit myocardium.
The muscarinic receptor-binding properties of a series of muscarinic drugs were compared with their effects on adenylate cyclase in membranes of the rabbit myocardium. When measured by competitive inhibition of [3H]-N-methylscopolamine binding, the competition curves of the various agonists were adequately described by the ternary complex model. This model assumes that the receptor can bind reversibly with a guanine nucleotide binding protein in the membrane and that the affinity of the agonist for the receptor-guanine nucleotide-binding protein complex is higher than that for the free receptor. A satisfactory fit of the ternary complex model to the data could only be achieved assuming that very little receptor is precoupled with the guanine nucleotide-binding protein in the absence of agonist. There was good agreement between the efficacy of each agonist as measured by inhibition of adenylate cyclase and the estimate of the positive cooperativity between the binding of the agonist receptor complex and the guanine nucleotide-binding protein. Guanosine 5'-triphosphate (0.1 mM) had no significant effect on the binding of [3H]N-methylscopolamine but caused an increase in the concentration of the various agonists required for half-maximal receptor occupancy. There was good correlation between efficacy as measured by inhibition of adenylate cyclase and the influence of guanosine 5'-triphosphate on binding properties.


Quality of care for remote orthopaedic consultations using telemedicine: a randomised controlled trial.
Decentralised services using outreach clinics or modern technology are methods to reduce both patient transports and costs to the healthcare system. Telemedicine consultations via videoconference are one such modality. Before new technologies are implemented, it is important to investigate both the quality of care given and the economic impact from the use of this new technology. The aim of this clinical trial was to study the quality of planned remote orthopaedic consultations by help of videoconference. We performed a randomised controlled trial (RCT) with two parallel groups: video-assisted remote consultations at a regional medical centre (RMC) as an intervention versus standard consultation in the orthopaedic outpatient clinic at the University Hospital of North Norway (UNN) as a control. The participants were patients referred to or scheduled for a consultation at the orthopaedic outpatient clinic. The orthopaedic surgeons evaluated each consultation they performed by completing a questionnaire. The primary outcome measurement was the difference in the sum score calculated from this questionnaire, which was evaluated by the non-inferiority of the intervention group. The study design was based on the intention to treat principle. Ancillary analyses regarding complications, the number of consultations per patient, operations, patients who were referred again and the duration of consultations were performed. Four-hundred patients were web-based randomised. Of these, 199 (98 %) underwent remote consultation and 190 (95 %) underwent standard consultation. The primary outcome, the sum score of the specialist evaluation, was significantly lower (i.e. 'better') at UNN compared to RMC (1.72 versus 1.82, p = 0.0030). The 90 % confidence interval (CI) for the difference in score (0.05, 0.17) was within the non-inferiority margin. The orthopaedic surgeons involved evaluated 98 % of the video-assisted consultations as 'good' or 'very good'. In the ancillary analyses, there was no significant difference between the two groups. This study supports the argument that it is safe to offer video-assisted consultations for selected orthopaedic patients. We did not find any serious events related to the mode of consultation. Further assessments of the economic aspects and patient satisfaction are needed before we can recommend its wider application. ClinicalTrials.gov identifier: NCT00616837.


TDP-43 loss-of-function causes neuronal loss due to defective steroid receptor-mediated gene program switching in Drosophila.
TDP-43 proteinopathy is strongly implicated in the pathogenesis of amyotrophic lateral sclerosis and related neurodegenerative disorders. Whether TDP-43 neurotoxicity is caused by a novel toxic gain-of-function mechanism of the aggregates or by a loss of its normal function is unknown. We increased and decreased expression of TDP-43 (dTDP-43) in Drosophila. Although upregulation of dTDP-43 induced neuronal ubiquitin and dTDP-43-positive inclusions, both up- and downregulated dTDP-43 resulted in selective apoptosis of bursicon neurons and highly similar transcriptome alterations at the pupal-adult transition. Gene network analysis and genetic validation showed that both up- and downregulated dTDP-43 directly and dramatically increased the expression of the neuronal microtubule-associated protein Map205, resulting in cytoplasmic accumulations of the ecdysteroid receptor (EcR) and a failure to switch EcR-dependent gene programs from a pupal to adult pattern. We propose that dTDP-43 neurotoxicity is caused by a loss of its normal function.


A case of Crohn's disease involving the gallbladder.
Crohn's disease is well known to affect any part of the gastrointestinal tract including the oral cavity and anus. Various extraintestinal complications have been reported in Crohn's disease, but extraintestinal involvement characterized by granulomatous lesions is uncommon. Here, we have reported a case about the involvement of the gallbladder in Crohn's disease. A 33-year-old woman was diagnosed having panperitonitis due to intestinal perforation and cholecystitis. The patient was moved to the surgical service for an emergency operation. On the resected specimen, there was a broad longitudinal ulcer at the mesenteric side. The mucosa of the gallbladder was nodular and granular, and the wall was thickened. The surface epithelium of the gallbladder was partially eroded and pyloric gland metaplasia was observed focally. Rokitansky-Aschoff sinuses were also present. From the lamina propria to the subserosal layer, there were several well-formed epithelioid cell granulomas, which were the non-caseating sarcoidal type different from the foreign-body and xanthomatous granulomas. Periodic-acid Schiff and acid fast stains revealed no organism within the granulomas. Lymphoid aggregates were present throughout the gallbladder wall. Sections from the resected ileum showed typical features of the Crohn's disease. When cholecystectomy is performed in a patient with Crohn's disease, the possibility of gallbladder involvement should be carefully examined by histopathological tests.


A comparison between 4 and 6 implants in the maxillary posterior region to support an overdenture; 1-year results from a randomized controlled trial.
To compare the treatment outcome of 4 vs. 6 bar-connected implants in the posterior region of the maxilla to support an overdenture during a 1-year follow-up period. Sixty-six edentulous patients with an insufficient amount of bone volume in the maxilla to place implants were asked to participate in this study. Randomization assigned patients to either 4 or 6 implants. In all patients, a maxillary sinus floor elevation procedure with bone from the iliac crest was performed, and after a 3-month healing period, 4 or 6 dental implants were inserted in the maxillary posterior region in a one-stage procedure. After 3 months of osseointegration, a bar-supported overdenture was constructed. Implant survival, overdenture survival, clinical scores, peri-implant bone height changes and patients' satisfaction were assessed. Study analysis was performed according a non-inferiority design. All patients completed the one-year follow-up. After a functional period of 1-year, implant survival was 100% in the 4 implants group and 99.5% in the 6 implants group. Overdenture survival was 100% in both groups. Mean clinical scores were very low and did not significantly differ between groups. Mean marginal bone resorption was 0.35 ± 0.31 mm and 0.46 ± 0.34 mm in the 4 and 6 implants group, respectively. Patients' satisfaction improved significantly in both groups, but did not differ between groups. A bar-supported overdenture on 4 implants in the posterior maxillary region is not inferior to an overdenture supported by 6 bar-connected dental implants.


The Physiological Effects of Dandelion (Taraxacum Officinale) in Type 2 Diabetes.
The tremendous rise in the economic burden of type 2 diabetes (T2D) has prompted a search for alternative and less expensive medicines. Dandelion offers a compelling profile of bioactive components with potential anti-diabetic properties. The Taraxacum genus from the Asteraceae family is found in the temperate zone of the Northern hemisphere. It is available in several areas around the world. In many countries, it is used as food and in some countries as therapeutics for the control and treatment of T2D. The anti-diabetic properties of dandelion are attributed to bioactive chemical components; these include chicoric acid, taraxasterol (TS), chlorogenic acid, and sesquiterpene lactones. Studies have outlined the useful pharmacological profile of dandelion for the treatment of an array of diseases, although little attention has been paid to the effects of its bioactive components on T2D to date. This review recapitulates previous work on dandelion and its potential for the treatment and prevention of T2D, highlighting its anti-diabetic properties, the structures of its chemical components, and their potential mechanisms of action in T2D. Although initial research appears promising, data on the cellular impact of dandelion are limited, necessitating further work on clonal β-cell lines (INS-1E), α-cell lines, and human skeletal cell lines for better identification of the active components that could be of use in the control and treatment of T2D. In fact, extensive in-vitro, in-vivo, and clinical research is required to investigate further the pharmacological, physiological, and biochemical mechanisms underlying the effects of dandelion-derived compounds on T2D.


Ab initio study of the radiation pressure on dielectric and magnetic media.
The Maxwell stress tensor and the distributed Lorentz force are applied to calculate forces on lossless media and are shown to be in excellent agreement. From the Maxwell stress tensor, we derive analytical formulae for the forces on both a half-space and a slab under plane wave incidence. It is shown that a normally incident plane wave pushes the slab in the wave propagation direction, while it pulls the half-space toward the incoming wave. Zero tangential force is derived at a boundary between two lossless media, regardless of incident angle. The distributed Lorentz force is applied to the slab in a direct way, while the half-space is dealt with by introducing a finite conductivity. In this regard, we show that the ohmic losses have to be properly accounted for, otherwise differing results are obtained. This contribution, together with a generalization of the formulation to magnetic materials, establishes the method on solid theoretical grounds. Agreement between the two methods is also demonstrated for the case of a 2-D circular dielectric particle.


Family relationships and troubled sleep among U.S. adults: examining the influences of contact frequency and relationship quality.
Sleep is essential for health and daily functioning, and social relationships may be a key social factor influencing sleep, yet sleep has been understudied in the literature on social relationships and health. This study used data from the National Survey of Midlife Development in the United States to examine associations between troubled sleep and family contact, social support, and strain. Results show that having strained family relationships is associated with more troubled sleep, while supportive family relationships are associated with less troubled sleep. Family strain is more consequential for sleep than support, and sleep troubles are greatest when family relationships are highly strained and provide inadequate emotional support. Family strain is also more harmful to sleep among individuals who are in frequent contact with family members. These findings underscore the importance of focusing on both negative and positive aspects of relationships and highlight the significance of family relationships for sleep.


Diagnosis and treatment of the hemiplegic patient with brachial plexus injury.
Brachial plexus injury was observed as a complication in 5 of 12 hemiplegic patients admitted over a 5-week period to an inpatient unit of the Rehabilitation Institute of Chicago. These patients exhibited unusual patterns of muscle atrophy and return of function in the impaired upper extremity. Occupational therapists may play an important part in the diagnosis and treatment of this complication of hemiplegia by promptly recognizing its subtle clinical signs and instituting appropriate therapy. Electromyography may be recommended to confirm this diagnosis. The treatment of choice is to maintain correct positioning of the limb both day and night, to use facilitation techniques for specific muscles in order to prevent atrophy, and to maintain passive range of motion as much as possible. Prevention of brachial plexus injury depends largely on the education of patient, family, and staff as to the potential hazards to a frail extremity that has no protective responses.


Can green space quantity and quality help prevent postpartum weight gain? A longitudinal study.
Postpartum weight gain is a well-known challenge for many mothers, but associations with green space quantity and quality have not been investigated. This longitudinal study used data on 3843 mothers living in Australia tracked biennially for 15 years post partum from 2004 onwards. Multilevel growth curve models adjusted for confounding were used to examine the patterning of body mass index (BMI) in relation to green space quantity, measured by percentage land use, and green space quality, measured by self-report. Two-way interaction terms were fitted to investigate time-contingent associations between BMI and green space. Compared with mothers in areas with ≤5% green space, adjusted BMI coefficients were -0.43 kg/m2 (SE 0.37), -0.69 kg/m2 (SE 0.32) -0.86 kg/m2 (SE 0.33) and -0.80 kg/m2 (SE 0.41) among mothers in areas with 6%-10%, 11%-20%, 21%-40% and ≥41% green space, respectively. There were no independent associations between BMI and green space quality. Evidence suggested mothers living in areas with 21%-40% green space had the lowest BMI, whether they agreed that local parks were good quality (-0.89 kg/m2 (SE 0.34)) or not (-0.93 kg/m2 (SE 0.35)). Mothers in the greenest areas only had statistically significantly lower BMI if they perceived local parks as high quality (-0.89 kg/m2 (SE 0.41)). There was limited evidence that these associations varied with respect to the number of years post partum. These findings may suggest that urban greening strategies to achieve a threshold of at least 21% or more green space in an area may help reduce, but not fully prevent postpartum weight gain. Potential mechanisms warrant investigation.


Lipid screening: is it enough to measure total cholesterol concentration?
To determine whether measurement of total cholesterol concentration is sufficient to identify most patients at lipoprotein mediated risk of coronary heart disease without measurement of triglyceride and high density lipoprotein (HDL) cholesterol concentrations. Cross sectional screening programme. Six general practices in Oxfordshire. 1901 Men and 2068 women aged 25-59. Cardiovascular risk as assessed by fasting venous plasma concentrations of total cholesterol, triglyceride, and HDL cholesterol. 2931 Patients (74% of those screened) had a total cholesterol concentration of less than 6.5 mmol/l. If the triglyceride concentration had not been measured in these patients isolated hypertriglyceridaemia (greater than or equal to 2.3 mmol/l) would have remained undetected in 185. Among these 185 patients, however, 123 were overweight or obese and only 18 (0.6% of those screened) had an increased risk associated with both a raised triglyceride concentration (greater than or equal to 2.3 mmol/l) and a low HDL cholesterol concentration (less than 0.9 mmol/l). Conversely, in the 790 patients with predominant hypercholesterolaemia (cholesterol concentration greater than or equal to 6.5 mmol/l and triglyceride concentration less than 2.3 mmol/l) measurement of HDL cholesterol concentration showed that 348 (9% of those screened) had only a moderately increased risk with a ratio of total to HDL cholesterol of less than 4.5 and 104 had a low risk with a ratio of less than 3.5. Fasting triglyceride and HDL cholesterol concentrations identify few patients at increased risk of coronary heart disease if the total cholesterol concentration is less than 6.5 mmol/l. HDL cholesterol and triglyceride concentrations should, however, be measured in patients with a total cholesterol concentration exceeding this value. Total cholesterol concentration alone may overestimate risk in a considerable number of these patients, and measurement of HDL cholesterol concentration allows a more precise estimate of risk. Measurement of the triglyceride concentration is required to characterise the lipoprotein abnormality. A patient should not be started on a drug that lowers lipid concentrations without having had a full lipoprotein assessment including measurement of HDL cholesterol concentration.


Healthfulness and nutritional composition of Canadian prepackaged foods with and without sugar claims.
The objective of this study was to evaluate differences in calories, nutrient content, overall healthfulness, and use of sweetener ingredients between products with and without sugar claims. Consumers assume products with sugar claims are healthier and lower in calories. It is therefore important claims be found on comparatively healthier items. This study is a cross-sectional analysis of the University of Toronto's 2013 Food Label Database. Subcategories where at least 5% of products (and n ≥ 5) carried a sugar claim were included (n = 3048). Differences in median calorie content, nutrient content, and overall healthfulness, using the Food Standards Australia/New Zealand Nutrient Profiling Scoring criterion, between products with and without sugar claims, were determined. Proportion of products with and without claims that had excess free sugar levels (≥10% of calories from free sugar) and that contained sweeteners was also determined. Almost half (48%) of products with sugar claims contained excess free sugar, and a greater proportion contained sweeteners than products without such claims (30% vs 5%, χ2 = 338.6, p < 0.0001). Overall, products with sugar claims were "healthier" and had lower median calorie, free sugar, total sugar, and sodium contents than products without claims. At the subcategory level, reductions in free sugar contents were not always met with similar reductions in calorie contents. This study highlights concerns with regards to the nutritional composition of products bearing sugar claims. Findings can support educational messaging to assist consumer interpretation of sugar claims and can inform changes in nutrition policies, for example, permitting sugar claims only on products with calorie reductions and without excess free sugar.


A new combined strategy to implement a community occupational therapy intervention: designing a cluster randomized controlled trial.
Even effective interventions for people with dementia and their caregivers require specific implementation efforts. A pilot study showed that the highly effective community occupational therapy in dementia (COTiD) program was not implemented optimally due to various barriers. To decrease these barriers and make implementation of the program more effective a combined implementation (CI) strategy was developed. In our study we will compare the effectiveness of this CI strategy with the usual educational (ED) strategy. In this cluster randomized, single-blinded, controlled trial, each cluster consists of at least two occupational therapists, a manager, and a physician working at Dutch healthcare organizations that deliver community occupational therapy. Forty-five clusters, stratified by healthcare setting (nursing home, hospital, mental health service), have been allocated randomly to either the intervention group (CI strategy) or the control group (ED strategy). The study population consists of the professionals included in each cluster and community-dwelling people with dementia and their caregivers. The primary outcome measures are the use of community OT, the adherence of OTs to the COTiD program, and the cost effectiveness of implementing the COTiD program in outpatient care. Secondary outcome measures are patient and caregiver outcomes and knowledge of managers, physicians and OTs about the COTiD program. Implementation research is fairly new in the field of occupational therapy, making this a unique study. This study does not only evaluate the effects of the CI-strategy on professionals, but also the effects of professionals' degree of implementation on client and caregiver outcomes. NCT01117285.


Oxygen is a factor determining in vitro tissue assembly: Effects on attachment and spreading of hepatocytes.
Many recent studies related to the development of bioartificial liver devices have utilized hepatocytes cultured within devices of various geometries. Because hepatocytes are anchorage-dependent cells, they need to attach and spread onto the extracellular matrix to be able to function, a process that requires energy. Thus, it is important to deliver enough oxygen to hepatocytes contained within bioartificial liver devices during the early phase of cellular organization while the cells interact with the extracellular matrix. In this study, we investigated the effect of oxygen on the attachment and spreading of hepatocytes. Increasing the gas phase oxygen from 0 to 160 mmHg resulted in an increase in the percentage of cells attaching from 43.0 +/- 5.8% to 103.6 +/- 29%, 1 h after seeding. In a similar manner, increasing the gas phase oxygen from 0 to 160 mmHg resulted in an increase of the projected surface area from 310 +/- 35 to 827 +/- 127 mum(2), 24 h after seeding. Furthermore, the partial pressure of oxygen at the cell level was estimated using a diffusion-reaction model. The model indicated that a cell surface oxygen partial pressure of 0.064 mmHg was required for the half-maximal (K(m) (a)) attachment of hepatocytes to collagen-based substrate. On the other hand, the K(m) (s) value of the spreading process was predicted to be 0.13 mmHg. The results of this study demonstrate the importance of oxygen during the initial stages of attachment and spreading of hepatocytes, and it has important implications in the design of hepatocyte-based bioartificial liver devices. (c) 1994 John Wiley & Sons, Inc.


HIV testing history and preferences for future tests among gay men, bisexual men and other MSM in England: results from a cross-sectional study.
The British HIV Association's (BHIVA) testing guidelines recommend men who have sex with men (MSM) test annually or more frequently if ongoing risk is present. We identify which groups of MSM in England are less likely to have tested for HIV and their preferences for future tests by testing model, in order to inform health promotion programmes. Data come from the Gay Men's Sex Survey 2014, a cross-sectional survey of MSM, aged 16 years or older and living in the UK. Only men who did not have diagnosed HIV and were living in England were included in this analysis. We used logistic regression models to understand how social determinants of health were associated with not testing for HIV in the past 12 months, and never having tested. We then cross-tabulated preferred testing location by demographic characteristics. Younger men, older men and men who were not gay identified were least likely to have tested for HIV. Higher educational attainment, migrancy, Black ethnicity and being at higher of risk were associated with greater levels of HIV testing. Men who were less likely to have tested for HIV preferred a wider range of options for future HIV testing. If the BHIVA's HIV testing policy of 2008 was used to guide testing priorities among MSM focus would be on increasing the rate of annual testing among MSM at less risk of HIV (ie, younger men, older men and non-gay identified MSM). Instead the promotion of more frequent testing among the groups most at risk of infection should be prioritised in order to reduce the time between infection and diagnosis.


Do we remember templates better so that we can reject distractors better?
Feature Integration Theory proposed that attention shifted between target-like representations in our visual field. However, the nature of the representations that determined what was target like received less specification than the nature of the attention shifts. In recent years, visual search research has focused on the nature of the memory representations that we use to guide our shifts of attention. Sensitive measures of memory quality indicate that the template representations are remembered better than other, merely maintained, memories. Here we tested the hypothesis that we prepare for difficult search tasks by storing a higher fidelity target representation in working memory than we do when preparing for an easy search task. To test this hypothesis, we explicitly tested participants' memory of the target color they searched for (i.e., the attentional template) versus another memory that was not used to guide attention (i.e., an accessory representation) following blocks of searches with easy-to-find targets (i.e., distractors were homogeneously colored) to blocks of searches with hard-to-find targets (i.e., distractors were heterogeneously colored). Although homogeneous-distractor searches required minimal precision for distractor rejection, we found that templates were still remembered better than accessories, just like we found in a heterogeneous-distractor search. As a consequence, we suggest that stronger memories for templates likely reflects the need to decide whether new perceptual inputs match the template, and not an attempt to create a better template representation in anticipation of difficult searches.


An activating Gs alpha mutation is present in fibrous dysplasia of bone in the McCune-Albright syndrome.
McCune-Albright syndrome (MAS) is a sporadic disease characterized by polyostotic fibrous dysplasia, café-au-lait spots, and multiple endocrinopathies. The etiology of fibrous dysplasia is unknown. Activating mutations of codon 201 in the gene encoding the alpha-subunit of Gs, the G-protein that stimulates adenylyl cyclase, have been found in all affected MAS tissues that have been studied. Initial attempts to amplify DNA from decalcified paraffin-embedded bone specimens from MAS patients were unsuccessful. Therefore, we analyzed DNA from frozen surgical bone specimens from five MAS patients using polymerase chain reaction and allele-specific oligonucleotide hybridization. Most of the cells in four specimens of dysplastic bone contained a heterozygous mutation encoding substitution of Arg201 of Gs alpha with His, but the mutation was barely detectable in peripheral blood specimens from the patients. Only a small amount of mutant allele was detected in a specimen of normal cortical bone from the fifth patient, although this patient had a high proportion of mutation in other, affected tissues. The mosaic distribution of mutant alleles is consistent with an embryological somatic cell mutation of the Gs alpha gene in MAS. The presence of an activating mutation of Gs alpha in osteoblastic progenitor cells may cause them to exhibit increased proliferation and abnormal differentiation, thereby producing the lesions of fibrous dysplasia.


Anti-SARS-CoV-2 IgG responses are powerful predicting signatures for the outcome of COVID-19 patients.
The COVID-19 global pandemic is far from ending. There is an urgent need to identify applicable biomarkers for early predicting the outcome of COVID-19. Growing evidences have revealed that SARS-CoV-2 specific antibodies evolved with disease progression and severity in COIVD-19 patients. We assumed that antibodies may serve as biomarkers for predicting the clinical outcome of hospitalized COVID-19 patients on admission. By taking advantage of a newly developed SARS-CoV-2 proteome microarray, we surveyed IgG responses against 20 proteins of SARS-CoV-2 in 1034 hospitalized COVID-19 patients on admission and followed till 66 days. The microarray results were further correlated with clinical information, laboratory test results and patient outcomes. Cox proportional hazards model was used to explore the association between SARS-CoV-2 specific antibodies and COVID-19 mortality. Nonsurvivors (n = 955) induced higher levels of IgG responses against most of non-structural proteins than survivors (n = 79) on admission. In particular, the magnitude of IgG antibodies against 8 non-structural proteins (NSP1, NSP4, NSP7, NSP8, NSP9, NSP10, RdRp, and NSP14) and 2 accessory proteins (ORF3b and ORF9b) possessed significant predictive power for patient death, even after further adjustments for demographics, comorbidities, and common laboratory biomarkers for disease severity (all with p trend < 0.05). Additionally, IgG responses to all of these 10 non-structural/accessory proteins were also associated with the severity of disease, and differential kinetics and serum positive rate of these IgG responses were confirmed in COVID-19 patients of varying severities within 20 days after symptoms onset. The area under curves (AUCs) for these IgG responses, determined by computational cross-validations, were between 0.62 and 0.71. Our findings might have important implications for improving clinical management of COVID-19 patients.


4,4'-Di-tert-butyl-6-(1H-tetrazol-5-yl)-2,2'-bipyridine: modification of a highly selective N-donor ligand for the separation of trivalent actinides from lanthanides.
In the present work, the complexation and extraction behaviour of 4,4'di-tert-butyl-6-(1H-tetrazol-5-yl)-2,2'-bipyridine (HN4tbubipy) towards trivalent actinides (An(iii)) and lanthanides (Ln(iii)) is studied by spectroscopic methods, liquid-liquid extraction, and quantum chemical calculations. The ligand synthesis of HN4tbubipy as well as its application in coordination chemistry of the 4f elements is described. Reaction of HN4tbubipy with [Ln(NO3)3·6H2O] (Ln = Sm, Eu) results in [H2N4tbubipy]+[Ln(N4tbubipy)(NO3)3(H2O)]-. Both compounds have been characterized by single crystal X-ray diffraction. The solubility of the ligand in different organic solvents is determined, showing a high solubility in MeOH which decreases with the lipophilicity of the solvent. The pKa = 2.4 ± 0.2 of HN4tbubipy in EtOH (4.4 vol% H2O) is determined by absorption spectrophotometry. The complexation of Cm(iii) and Eu(iii) with HN4tbubipy is studied by time resolved laser fluorescence spectroscopy (TRLFS). For both metal ions the formation of the complexes [M(N4tbubipy)n]3-n with n = 2, 3 (M = Cm(iii), Eu(iii)) is observed. Slightly higher conditional stability constants for Eu(iii) (log β'2(Eu(N4tbubipy)2+) = 8.9 ± 0.3, log β'3(Eu(N4tbubipy)3) = 12.7 ± 0.5), compared to Cm(iii) (log β'2(Cm(N4tbubipy)2+) = 8.5 ± 0.4 and log β'3(Cm(N4tbubipy)3) = 12.4 ± 0.6) are determined. Thus, the ligand has no preference for the complexation of An(iii) over Ln(iii). Additionally, no significant extraction of Am(iii) and Eu(iii) is observed in liquid-liquid extraction experiments due to protonation of the ligand at the experimental conditions. The experimental studies are supported by quantum chemical calculations of the free ligand and the [M(N4tbubipy)3] complexes (M = Cm(iii), Gd(iii)). The results are in excellent agreement with the experimental data and provide a deeper understanding of the complexation properties of HN4tbubipy.


Prenatal exposures to bisphenol A and di (2-ethylhexyl) phthalate disrupted seminiferous tubular development in growing male rats.
Endocrine-disrupting compounds (EDCs) are found in the environment due to their use in industrial and manufacturing activities. Exposure of the population to bisphenol A (BPA) and di (2-ethylhexyl) phthalate (DEHP) is significant because they are present in many consumer products. EDCs target the reproductive tract because they express high levels of steroid hormone receptors, which act as transcriptional factors to regulate reproductive development. In the present study, timed-pregnant Long-Evans female rats (n = 8-10) were administered BPA and DEHP by oral gavage at 2.5 or 25 μg/kg body weight and 5 or 50 μg/kg body weight, respectively. Exposures to chemicals were limited to the period between gestational days 12 and 21 followed by assessment of testicular development in male offspring in the postnatal period. Leydig cells and Sertoli cells are the two major somatic cells present in the testis. The 17β-hydroxysteroid dehydrogenase (17β-HSD) steroidogenic enzyme is a marker for Leydig cell maturation, whereas transferrin is a marker for Sertoli cell differentiation. At day 10 post-partum, testes were obtained from cohorts of control and chemical-exposed male rats and processed to measure 17β-HSD and transferrin expression levels in western blots. Compared to control, 17βHSD enzyme protein was increased in BPA-treated rats but levels were decreased in animals exposed to DEHP (P < 0.05). Transferrin protein was decreased in male rats exposed to both BPA and DEHP compared to control animals (P < 0.05). To assess qualitative cellular changes within the spermatogenic epithelium, testes were obtained from separate cohorts of male rats at 35 days of age and processed for histopathological analysis. Results showed that prenatal exposures of male rats to BPA and DEHP caused disruption of the spermatogenic epithelium evident as disorganization and atrophy of seminiferous tubules as well as desquamation of germ cells into the tubular lumen. Together, results from the present study support the view that developmental exposures to environmentally relevant levels of BPA and DEHP are associated with disruptions of testicular cell development, which have implications for endocrine and exocrine functions of testis.


Hair loss perception and symptoms of depression in female outpatients attending a general dermatology clinic.
Hair loss is a common complaint among women and may be associated with psychological disturbances. This study investigated dermatology patients with respect to the prevalence of complaints of hair loss and the association between these complaints and symptoms of depression. A cross-sectional study was conducted with female outpatients over 20 years of age with no hair-related disorders. Patients were asked about hair loss and were evaluated with respect to symptoms of depression. The following variables were also analyzed: age, dermatological diagnosis, comorbidities, medications, family history of alopecia, hair length, the number of hairs extracted in the hair pull test and the presence of hair thinning or seborrheic dermatitis. Data were compared by bivariate analysis and multiple logistic regressions between groups of women with and without hair loss. Of the 157 women interviewed, 54% reported hair loss and 29% reported at least two key symptoms of depression. The median (IQD) age of the women was 51 (20) years. Complaints of hair loss were associated with the presence of symptoms of depression even following adjustment for the other covariates (p=0.02; OR=2.79; 95%CI: 1.18-6.61). Statistically significant differences were also found between the patients with and without hair loss with respect to: age (p=0.03), family history of alopecia (p<0.01), hair length (p=0.01), number of hairs extracted in the hair pull test (p=0.02), hair thinning (p<0.01), seborrheic dermatitis (p<0.01) and problems with personal relationships (p=0.04). Hair-related disorders are a common topic of interest in this population. The significant association found between this complaint and symptoms of depression indicates a relationship between the perception of hair loss and the psychological well being of the women evaluated. Complaints of hair loss were common and were associated with a greater prevalence of symptoms of depression among adult female outpatients at a public dermatology clinic.


Relation between dynamics, activity and thermal stability within the cholinesterase family.
Incoherent neutron scattering is one of the most powerful tools for studying dynamics in biological matter. Using the cold neutron backscattering spectrometer IN16 at the Institut Laue Langevin (ILL, Grenoble, France), temperature dependence of cholinesterases' dynamics (human butyrylcholinesterase from plasma: hBChE; recombinant human acetylcholinesterase: hAChE and recombinant mouse acetylcholinesterase: mAChE) was examined using elastic incoherent neutron scattering (EINS). The dynamics was characterized by the averaged atomic mean square displacement (MSD), associated with the sample flexibility at a given temperature. We found MSD values of hAChE above the dynamical transition temperature (around 200K) larger than for mAChE and hBChE, implying that hAChE is more flexible than the other ChEs. Activation energies for thermodynamical transition were extracted through the frequency window model (FWM) (Becker et al. 2004) [1] and turned out to increase from hBChE to mAChE and finally to hAChE, inversely to the MSDs relations. Between 280 and 316K, catalytic studies of these enzymes were carried out using thiocholine esters: at the same temperature, the hAChE activity was systematically higher than the mAChE or hBChE ones. Our results thus suggest a strong correlation between dynamics and activity within the ChE family. We also studied and compared the ChEs thermal inactivation kinetics. Here, no direct correlation with the dynamics was observed, thus suggesting that relations between enzyme dynamics and catalytic stability are more complex. Finally, the possible relation between flexibility and protein ability to grow in crystals is discussed.


Making health insurance cost-sharing clear to consumers: challenges in implementing health reform's insurance disclosure requirements.
The Affordable Care Act calls for a new health insurance disclosure form, called the Summary of Benefits and Coverage, which uses a fixed layout and standard terms and definitions to allow consumers to compare health insurance plans and understand terms of coverage. This brief reports on findings from a Consumers Union study that examined consumers' initial reactions to the form. Testing revealed that consumers were able to use the forms to make hypothetical choices among health plans. However, the study also found deep-seated confusion and lack of confidence with respect to health plan cost-sharing. These findings have significant implications for any venue providing comparative displays of health insurance information, like the future state exchanges, and for policies that rely on the ability of consumers to make informed health insurance purchasing decisions, such as "consumer-driven health care" policies.


Phylogeny of the insulin-like growth factors (IGFs) and receptors: a molecular approach.
The IGFs (IGF-I and IGF-II) are essential for normal mammalian growth and development. Their actions are mediated primarily by their interactions with the type I IGF receptor (IGF-I receptor), a transmembrane tyrosine kinase. The ligands and the IGF-I receptor are structurally related to insulin and to the insulin receptor, respectively. Analysis of evolutionary conservation has often provided insights into essential regions of molecules such as hormones and their receptors. The genes for insulin and IGFs have been partially characterized in a number of vertebrate species extending evolutionarily from humans as far back as fish. The sequences of the exons encoding the mature insulin and IGF peptides are highly conserved among vertebrate species, and IGF-I-like molecules are found in species whose origins extend back as much as 550 million years. The insulin receptor is also highly conserved in vertebrate species, and an insulin-receptor-like molecule has been characterized in Drosophila. In contrast, IGF-I receptors have only been characterized in mammalian species and partially studied in Xenopus, in which the tyrosine kinase domain is highly conserved. Studies are presently being undertaken to analyze in more detail the regulation of the genes encoding this important family of growth factors and the structure/function relationships in the gene products themselves.


Rapid evolution of parasite resistance in a warmer environment: insights from a large scale field experiment.
Global climate change is expected to have major effects on host-parasite dynamics, with potentially enormous consequences for entire ecosystems. To develop an accurate prognostic framework, theoretical models must be supported by empirical research. We investigated potential changes in host-parasite dynamics between a fish parasite, the eyefluke Diplostomum baeri, and an intermediate host, the European perch Perca fluviatilis, in a large-scale semi-enclosed area in the Baltic Sea, the Biotest Lake, which since 1980 receives heated water from a nuclear power plant. Two sample screenings, in two consecutive years, showed that fish from the warmer Biotest Lake were now less parasitized than fish from the Baltic Sea. These results are contrasting previous screenings performed six years after the temperature change, which showed the inverse situation. An experimental infection, by which perch from both populations were exposed to D. baeri from the Baltic Sea, revealed that perch from the Baltic Sea were successfully infected, while Biotest fish were not. These findings suggest that the elevated temperature may have resulted, among other outcomes, in an extremely rapid evolutionary change through which fish from the experimental Biotest Lake have gained resistance to the parasite. Our results confirm the need to account for both rapid evolutionary adaptation and biotic interactions in predictive models, and highlight the importance of empirical research in order to validate future projections.


Effects of combined administration of low dose atorvastatin and vitamin E on inflammatory markers and endothelial function in patients with heart failure.
Heart failure has been associated with impaired endothelial function, increased inflammatory process and elevated oxidative stress status. Both statins and vitamin E separately improve endothelial function in patients with hypercholesterolemia and/or advanced atherosclerosis. To evaluate the effect of atorvastatin alone or in combination with vitamin E on endothelial function and serum levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and vascular cells adhesion molecule (sVCAM-1) in patients with ischemic heart failure. Thirty-eight male patients with ischemic cardiomyopathy were randomly divided into three groups and received either atorvastatin 10 mg/day (n = 14), a combination of atorvastatin 10 mg/day plus vitamin E 400 IU/day (n = 12), or no statin or antioxidant treatment (n=12, controls) for 4 weeks. Forearm blood flow (FBF) was measured using venous occlusion strain-gauge plethysmography. Forearm vasodilatory response to reactive hyperemia (RH%) or to nitrate (NTG%) was defined as the percent change of FBF from rest to the maximum flow during reactive hyperemia or after nitrate administration, respectively. RH% was significantly improved in both the atorvastatin-treated (p < 0.01) and atorvastatin plus vitamin E groups (p < 0.05), but the increase was significantly higher in the atorvastatin-treated group (p < 0.05). Serum levels of IL-6, TNF-alpha and sVCAM-1 were decreased in the atorvastatin-treated group (p < 0.05 for all), but remained unaffected in the other two groups (p = NS for all). Low dose atorvastatin treatment improves endothelial function and reduces the expression of proinflammatory cytokines and adhesion molecules in patients with ischemic heart failure, an effect partly depressed by vitamin E.


Emergence of azole resistance in Aspergillus fumigatus and spread of a single resistance mechanism.
Resistance to triazoles was recently reported in Aspergillus fumigatus isolates cultured from patients with invasive aspergillosis. The prevalence of azole resistance in A. fumigatus is unknown. We investigated the prevalence and spread of azole resistance using our culture collection that contained A. fumigatus isolates collected between 1994 and 2007. We investigated the prevalence of itraconazole (ITZ) resistance in 1,912 clinical A. fumigatus isolates collected from 1,219 patients in our University Medical Centre over a 14-y period. The spread of resistance was investigated by analyzing 147 A. fumigatus isolates from 101 patients, from 28 other medical centres in The Netherlands and 317 isolates from six other countries. The isolates were characterized using phenotypic and molecular methods. The electronic patient files were used to determine the underlying conditions of the patients and the presence of invasive aspergillosis. ITZ-resistant isolates were found in 32 of 1,219 patients. All cases were observed after 1999 with an annual prevalence of 1.7% to 6%. The ITZ-resistant isolates also showed elevated minimum inhibitory concentrations of voriconazole, ravuconazole, and posaconazole. A substitution of leucine 98 for histidine in the cyp51A gene, together with two copies of a 34-bp sequence in tandem in the gene promoter (TR/L98H), was found to be the dominant resistance mechanism. Microsatellite analysis indicated that the ITZ-resistant isolates were genetically distinct but clustered. The ITZ-sensitive isolates were not more likely to be responsible for invasive aspergillosis than the ITZ-resistant isolates. ITZ resistance was found in isolates from 13 patients (12.8%) from nine other medical centres in The Netherlands, of which 69% harboured the TR/L98H substitution, and in six isolates originating from four other countries. Azole resistance has emerged in A. fumigatus and might be more prevalent than currently acknowledged. The presence of a dominant resistance mechanism in clinical isolates suggests that isolates with this mechanism are spreading in our environment.


Melanocortin receptors as targets in the treatment of obesity.
The central melanocortin system plays a critical role in energy homeostasis. It is well established that melanocortin-containing neurons are nutritionally regulated and that genetic alterations in the melanocortin system produce profound effects on food intake, energy expenditure, and body weight. Within the brain, melanocortin-producing neurons originate in the arcuate nucleus of the hypothalamus (ARC) and the nucleus of the solitary tract (NTS) in the brainstem and project to various nuclei modulating energy balance. A large body of pharmacological and genetic evidence implicates the central melanocortin 4 receptors (MC4Rs) in the effects of melanocortin peptides on ingestive behaviour, energy expenditure, and body weight. Preclinical studies with endogenous and synthetic melanocortin ligands demonstrate that they produce potent effects on food intake and energy expenditure. Clinical studies thus far have been somewhat less successful and have been hampered by the induction of side effects, which present obstacles to the development of successful therapeutic agents. However, various promising strategies are being pursued to overcome these limitations, including the synthesis of more selective and potent melanocortin analogs.