Determining readiness for a reablement approach to care in Australia: Development of a pre-employment questionnaire.
Reablement is described as a goal directed intervention with a view to maintain or improve the independence of clients through "doing with, rather than doing for". As a relatively new concept in care, lack of specific skills in reablement is not uncommon, however, intrinsic values aligned with reablement are beneficial to organisations providing care and support to clients. The aim of this study is to develop a pre-employment questionnaire to assess readiness for a reablement approach to care as a step towards developing a reablement culture across organisations. A questionnaire based on activities of daily living (ADL), developed by the research team in consultation with a local community-based care organisation, was completed by 166 staff members at a community-based care organisation in Tasmania across several timepoints during and following reablement education sessions in 2018. The scores from these questionnaires were utilised to develop a questionnaire appropriate for determining readiness for reablement prior to employment within the organisation. Over a period of twelve months, a total of 407 Reablement Readiness Questionnaires were submitted by staff, indicating high levels of willingness to engage in reablement prior to and following the education sessions. Exploratory factor analyses were performed, and the construct validity and internal consistency of the emerging factors were assessed. An exploratory factor analysis indicated that ADL, in relation to reablement, consist of three dimensions-Lifestyle, My body and Connecting. The corresponding questions were subsequently built into an online questionnaire for pre-employment use. This questionnaire also has potential benefit in recruitment of staff to other community service organisations, to ascertain their readiness for reablement prior to beginning a variety of different roles within client care and support.


[Human spongiform encephalopathies in the Netherlands].
Cases of Creutzfeldt Jakob disease (CJD) have been centrally registered in the Netherlands since 1997. Each year about 35 cases are reported. All patients with a possible, probable or definite diagnosis are registered in a European database. Recently, patients with a transfusion-transmitted form of CJD have been diagnosed in the United Kingdom. Such cases have not yet occurred in the Netherlands. It is important to continue to detect and centrally register patients with CJD.


Social density processes regulate the functioning and performance of foraging human teams.
Social density processes impact the activity and order of collective behaviours in a variety of biological systems. Much effort has been devoted to understanding how density of people affects collective human motion in the context of pedestrian flows. However, there is a distinct lack of empirical data investigating the effects of social density on human behaviour in cooperative contexts. Here, we examine the functioning and performance of human teams in a central-place foraging arena using high-resolution GPS data. We show that team functioning (level of coordination) is greatest at intermediate social densities, but contrary to our expectations, increased coordination at intermediate densities did not translate into improved collective foraging performance, and foraging accuracy was equivalent across our density treatments. We suggest that this is likely a consequence of foragers relying upon visual channels (local information) to achieve coordination but relying upon auditory channels (global information) to maximise foraging returns. These findings provide new insights for the development of more sophisticated models of human collective behaviour that consider different networks for communication (e.g. visual and vocal) that have the potential to operate simultaneously in cooperative contexts.


Compositional Analysis of Genetically Engineered GR2E "Golden Rice" in Comparison to That of Conventional Rice.
Compositional analyses were performed on samples of rice grain, straw, and derived bran obtained from golden rice event GR2E and near-isogenic control PSBRc82 rice grown at four locations in the Philippines during 2015 and 2016. Grain samples were analyzed for key nutritional components, including proximates, fiber, polysaccharides, fatty acids, amino acids, minerals, vitamins, and antinutrients. Samples of straw and bran were analyzed for proximates and minerals. The only biologically meaningful difference between GR2E and control rice was in levels of β-carotene and other provitamin A carotenoids in the grain. Except for β-carotene and related carotenoids, the compositional parameters of GR2E rice were within the range of natural variability of those components in conventional rice varieties with a history of safe consumption. Mean provitamin A concentrations in milled rice of GR2E can contribute up to 89-113% and 57-99% of the estimated average requirement for vitamin A for preschool children in Bangladesh and the Philippines, respectively.


In vivo confocal microscopy of conjunctiva-associated lymphoid tissue in healthy humans.
To investigate modifications with aging of the presence, distribution and morphologic features of conjunctiva-associated lymphoid tissue (CALT) in healthy human subjects using laser scanning in vivo confocal microscopy (IVCM). A total of 108 (age range, 17-75 years) subjects were enrolled. In vivo confocal microscopy of the tarsal and bulbar conjunctiva, and impression cytology (IC) with CD3 (intra-epithelial T-lymphocytes) and CD20 (intra-epithelial B-lymphocytes) antibody immunofluorescence staining were performed. The main outcomes were subepithelial lymphocyte density (LyD), follicular density (FD), and follicular area (FA). The secondary outcomes were follicular reflectivity (FR), and lymphocyte density (FLyD), and CD3 and CD20 positivity. Conjunctiva-associated lymphoid tissue was observed in all subjects (97% only superior and 3% in both superior and inferior tarsum). Lymphocyte density ranged from 7.8 to 165.8 cells/mm(2) (46.42 [18.37]; mean [SD]), FD from 0.5 to 19.4 follicles/mm(2) (5.3 [3.6]), and FA from 1110 to 96,280 mm(2) (26,440 [26,280]). All three parameters showed a highly significant inverse cubic relationship with age (P < 0.001); that is, in the first and last parameters a steep decline up to 35 years and above 65 years of age, with a plateau phase between these ages, whereas FA had a gradually decreasing rate of loss over the studied age range. CD3 and CD20 IC were consistent with these results. In vivo confocal microscopy was effective in revealing CALT and modifications these structures undergo with aging. Aging correlated with an involution of all parameters defining lymphoid structures. These modifications may account for the decrease of mucosal immune response and increase of ocular surface diseases in the elderly.


The effect of community based-academic partnerships on student knowledge about plastic surgery and interest in medicine.
Programs specific to plastic surgery are necessary to dispel common myths and increase interest in the field. In a previous publication by the authors, a community outreach program was developed for these reasons for middle school students. In the current study, we expanded on the previous research and collected objective data to assess students' initial interest in medicine and knowledge about plastic surgery, compared to their interest and knowledge afterward. The program previously developed by the authors was modified and performed for the students at various community outreach events and included a PowerPoint presentation, case didactics, and hands-on activities. A test about plastic surgery and questionnaire about interest in the medical field and becoming a doctor was given to each student before and after the program. One hundred seventy-nine students participated in the program from 2009 to 2013. The pretest mean score was 6.50 of 12 questions whereas the posttest mean score was 9.72 (P = <0.001). After participation in the program, 27% of students that answered "no" or "unsure" about interest in the medical field on the pretest changed their answer to "yes," on the posttest, and 17% of students that answered "no" or "unsure" about interest in becoming a doctor on the pretest changed their answer to "yes," on the posttest (P = <0.001). A plastic surgery community outreach program is beneficial in increasing students' interest in the field of medicine as a whole, and more specifically in the field of plastic surgery.


Biparental care is more than the sum of its parts: experimental evidence for synergistic effects on offspring fitness.
Despite an extensive body of theoretical and empirical literature on biparental cooperation, it is still unclear whether offspring fare equally, better or worse when receiving care by two parents versus a single parent. Some models predict that parents should withhold the amount of care they provide due to sexual conflict, thereby shifting as much of the workload as possible to their partner. This conflict should lead to offspring faring worse with two parents. Yet, other models predict that when parents care for their offspring together, their individual contributions can have synergistic (more than additive) effects on offspring fitness. Under this scenario, biparental cooperation should lead to offspring faring better with two parents. We address this fundamental question using a unique experimental design where we compared offspring fitness when the two parents worked together (biparental treatment) and when they worked separately (uniparental treatment), while keeping constant the amount of resources and number of offspring per parent across treatments. This made it possible to directly compare the biparental treatment to the sum of the male and female contributions in the uniparental treatment. Our main finding was that offspring grew larger and were more likely to survive to adulthood when reared by both parents than a single parent. This is the first empirical evidence for a synergistic effect of biparental cooperation on offspring fitness and could provide novel insights into the conditions favouring the evolution of biparental cooperation.


Human osteoblasts' proliferative responses to strain and 17beta-estradiol are mediated by the estrogen receptor and the receptor for insulin-like growth factor I.
The mechanism by which mechanical strain and estrogen stimulate bone cell proliferation was investigated using monolayer cultures of human osteoblastic TE85 cells and female human primary (first-passage) osteoblasts (fHOBs). Both cell types showed small but statistically significant dose-dependent increases in [3H]thymidine incorporation in response to 17beta-estradiol and to a single 10-minute period of uniaxial cyclic strain (1 Hz). In both cell types, the peak response to 17beta-estradiol occurred at 10(-8) - 10(-7) M and the peak response to strain occurred at 3500 microstrain ((mu)epsilon). Both strain-related and 17beta-estradiol-related increases in [3H]thymidine incorporation were abolished by the estrogen receptor (ER) modulator ICI 182,780 (10-8 M). Tamoxifen (10(-9) - 10(-8) M) increased [3H]thymidine incorporation in both cell types but had no effect on their response to strain. In TE85 cells, tamoxifen reduced the increase in [3H]thymidine incorporation associated with 17beta-estradiol to that of tamoxifen alone but had no such effect in fHOBs. In TE85 cells, strain increased medium concentrations of insulin-like growth factor (IGF) II but not IGF-I, whereas 17beta-estradiol increased medium concentrations of IGF-I but not IGF-II. Neutralizing monoclonal antibody (MNAb) to IGF-I (3 microg/ml) blocked the effects of 17beta-estradiol and exogenous truncated IGF-I (tIGF-I; 50 ng/ml) but not those of strain or tIGF-II (50 ng/ml). Neutralizing antibody to IGF-II (3 microg/ml) blocked the effects of strain and tIGF-II but not those of 17beta-estradiol or tIGF-I. MAb aIR-3 (100 ng/ml) to the IGF-I receptor blocked the effects on [3H]thymidine incorporation of strain, tIGF-II, 17beta-estradiol, and tIGF-I. HOBs and TE85 cells, act similarly to rat primary osteoblasts and ROS 17/2.8 cells in their dose-related proliferative responses to strain and 17beta-estradiol, both of which can be blocked by the ER modulator ICI 182,780. In TE85 cells (as in rat primaries and ROS 17/2.8 cells), the response to 17beta-estradiol is mediated by IGF-I, and the response to strain is mediated by IGF-II. Human cells differ from rat cells in that tamoxifen does not block their response to strain and reduces the response to 17beta-estradiol in TE85s but not primaries. In both human cell types (unlike rat cells) the effects of strain and IGF-II as well as estradiol and IGF-I can be blocked at the IGF-I receptor.


Comparing the impact of time displaced and biased precipitation estimates for online updated urban runoff models.
When an online runoff model is updated from system measurements, the requirements of the precipitation input change. Using rain gauge data as precipitation input there will be a displacement between the time when the rain hits the gauge and the time where the rain hits the actual catchment, due to the time it takes for the rain cell to travel from the rain gauge to the catchment. Since this time displacement is not present for system measurements the data assimilation scheme might already have updated the model to include the impact from the particular rain cell when the rain data is forced upon the model, which therefore will end up including the same rain twice in the model run. This paper compares forecast accuracy of updated models when using time displaced rain input to that of rain input with constant biases. This is done using a simple time-area model and historic rain series that are either displaced in time or affected with a bias. The results show that for a 10 minute forecast, time displacements of 5 and 10 minutes compare to biases of 60 and 100%, respectively, independent of the catchments time of concentration.


A double-blind randomized phase I clinical trial targeting ALVAC-HIV vaccine to human dendritic cells.
We conducted a novel pilot study comparing different delivery routes of ALVAC-HIV (vCP205), a canarypox vaccine containing HIV gene inserts: env, gag and pol. We explored the concept that direct ex vivo targeting of human dendritic cells (DC) would enhance the immune response compared to either conventional intramuscular or intradermal injections of the vaccine alone. Healthy HIV-1 uninfected volunteers were administered ALVAC-HIV or placebo by intramuscular injection (i.m.), intradermal injection (i.d.) or subcutaneous injection (s.q.) of autologous ex vivo transfected DC at months 0, 1, 3 and 6. All vaccine delivery routes were well tolerated. Binding antibodies were observed to both the ALVAC vector and HIV-1 gp160 proteins. Modest cellular responses were observed in 2/7 individuals in the DC arm and 1/8 in the i.m. arm as determined by IFN-γ ELISPOT. Proliferative responses were most frequent in the DC arm where 4/7 individuals had measurable responses to multiple HIV-1 antigens. Loading DC after maturation resulted in lower gene expression, but overall better responses to both HIV-1 and control antigens, and were associated with better IL-2, TNF-α and IFN-γ production. ALVAC-HIV delivered i.m., i.d. or s.q. with autologous ex vivo transfected DC proved to be safe. The DC arm was most immunogenic. Proliferative immune responses were readily detected with only modest cytotoxic CD8 T cell responses. Loading mature DC with the live viral vaccine induced stronger immune responses than loading immature DC, despite increased transgene expression with the latter approach. Volunteers who received the autologous vaccine loaded mature DC developed a broader and durable immune response compared to those vaccinated by conventional routes. ClinicalTrials.gov NCT00013572.


Prevalence of the CHEK2 R95* germline mutation.
While germline CHEK2 mutations have been linked to a moderately elevated cancer risk, to date, a limited number of such mutations have been identified. Recently, we reported a germline nonsense mutation (C283T; R95*), introducing an early stop-codon, in two Norwegian patients diagnosed with locally advanced breast cancer. Both patients were resistant to anthracycline therapy, resembling what has been observed for TP53 mutations. In the present study, we screened a large population based sample, including 3748 non-cancer individuals and 7081 incident cancer cases (breast cancer, n = 1717; prostate cancer n = 2501, lung cancer n = 1331 and colorectal cancer n = 1532), for the distribution of CHEK2 R95*. We found that 12 individuals (0.11 %) carried the R95* variant: 4 non-cancer individuals (0.11 %), 4 breast cancer cases (0.23 %), and 4 prostate cancer cases (0.16 %). Although the low number of observations precluded formal statistical assessment, our data may indicate an elevated risk for breast (OR: 2.19, 95 % CI: 0.55-8.75) and prostate cancer (OR: 1.5, 95 % CI: 0.36-6.00) associated with CHEK2 R95*. By mining international databanks, we found no individuals carrying the R95* mutation, indicating it to be restricted to the Norwegian population. We provide proof-of-concept that previously unknown CHEK2 germline mutations may be present in certain populations. Notably, germline mutations in tumours are in general missed by contemporary massive parallel sequencing strategies, since tumour mutations are usually filtered against the germline. The fact that the CHEK2 R95* mutation may be associated with resistance to anthracyclines in cancer patients emphasizes its possible clinical importance.


The origins and molecular evolution of SARS-CoV-2 lineage B.1.1.7 in the UK.
The first SARS-CoV-2 variant of concern (VOC) to be designated was lineage B.1.1.7, later labelled by the World Health Organization as Alpha. Originating in early autumn but discovered in December 2020, it spread rapidly and caused large waves of infections worldwide. The Alpha variant is notable for being defined by a long ancestral phylogenetic branch with an increased evolutionary rate, along which only two sequences have been sampled. Alpha genomes comprise a well-supported monophyletic clade within which the evolutionary rate is typical of SARS-CoV-2. The Alpha epidemic continued to grow despite the continued restrictions on social mixing across the UK and the imposition of new restrictions, in particular, the English national lockdown in November 2020. While these interventions succeeded in reducing the absolute number of cases, the impact of these non-pharmaceutical interventions was predominantly to drive the decline of the SARS-CoV-2 lineages that preceded Alpha. We investigate the only two sampled sequences that fall on the branch ancestral to Alpha. We find that one is likely to be a true intermediate sequence, providing information about the order of mutational events that led to Alpha. We explore alternate hypotheses that can explain how Alpha acquired a large number of mutations yet remained largely unobserved in a region of high genomic surveillance: an under-sampled geographical location, a non-human animal population, or a chronically infected individual. We conclude that the latter provides the best explanation of the observed behaviour and dynamics of the variant, although the individual need not be immunocompromised, as persistently infected immunocompetent hosts also display a higher within-host rate of evolution. Finally, we compare the ancestral branches and mutation profiles of other VOCs and find that Delta appears to be an outlier both in terms of the genomic locations of its defining mutations and a lack of the rapid evolutionary rate on its ancestral branch. As new variants, such as Omicron, continue to evolve (potentially through similar mechanisms), it remains important to investigate the origins of other variants to identify ways to potentially disrupt their evolution and emergence.


Interaction of the cholesteryl ester transfer protein I405V polymorphism with alcohol consumption in smoking and non-smoking healthy men, and the effect on plasma HDL cholesterol and apoAI concentration.
Three hundred and sixteen healthy Icelandic men and women were examined for the effect of the cholesteryl ester transfer protein (CETP) I405V polymorphism on plasma triglycerides, HDL cholesterol (HDLC) and apoAI concentration. Genotyping was performed using an allele specific oligomelting assay and the frequency of the V allele was 0.31 (95 CI for men 0.23-0.33 and for women 0.29-0.39). In women no significant difference was associated with the V405 genotype for any plasma lipid trait. However, men who were homozygous for the V405 allele had 9% higher apoAI and 14% higher HDLC levels (p < 0.05) than those homozygous for the common I405 allele. The genotype effect was seen only in the non-smokers (p = 0.07 and < 0.05, respectively), and in those consuming alcohol (p < 0.05 for both). Analysis of interaction between the environmental, life-style factors and genotype in men for the traits of HDLC and apoAI showed statistically significant interaction of the genotype only with alcohol consumption. The non-smoking men who reported alcohol consumption and who were homozygous for the CETP V405 allele had 16% higher plasma apoAI concentration than those who carried the I405 allele, and up to 20% higher apoAI level than smokers. On the basis of prospective studies carried out on the Icelandic population, non-smoking, alcohol-consuming men who are homozygous for the V405 allele could have from 32% to 40% lower risk of having a heart attack.


Efficacy and effectiveness of screen and treat policies in prevention of type 2 diabetes: systematic review and meta-analysis of screening tests and interventions.
To assess diagnostic accuracy of screening tests for pre-diabetes and efficacy of interventions (lifestyle or metformin) in preventing onset of type 2 diabetes in people with pre-diabetes. Systematic review and meta-analysis. Medline, PreMedline, and Embase. Study protocols and seminal papers were citation-tracked in Google Scholar to identify definitive trials and additional publications. Data on study design, methods, and findings were extracted onto Excel spreadsheets; a 20% sample was checked by a second researcher. Data extracted for screening tests included diagnostic accuracy and population prevalence. Two meta-analyses were performed, one summarising accuracy of screening tests (with the oral glucose tolerance test as the standard) for identification of pre-diabetes, and the other assessing relative risk of progression to type 2 diabetes after either lifestyle intervention or treatment with metformin. Empirical studies evaluating accuracy of tests for identification of pre-diabetes. Interventions (randomised trials and interventional studies) with a control group in people identified through screening. No language restrictions. 2874 titles were scanned and 148 papers (covering 138 studies) reviewed in full. The final analysis included 49 studies of screening tests (five of which were prevalence studies) and 50 intervention trials. HbA1c had a mean sensitivity of 0.49 (95% confidence interval 0.40 to 0.58) and specificity of 0.79 (0.73 to 0.84), for identification of pre-diabetes, though different studies used different cut-off values. Fasting plasma glucose had a mean sensitivity of 0.25 (0.19 to 0.32) and specificity of 0.94 (0.92 to 0.96). Different measures of glycaemic abnormality identified different subpopulations (for example, 47% : of people with abnormal HbA1c had no other glycaemic abnormality). Lifestyle interventions were associated with a 36% (28% to 43%) reduction in relative risk of type 2 diabetes over six months to six years, attenuating to 20% (8% to 31%) at follow-up in the period after the trails. HbA1c is neither sensitive nor specific for detecting pre-diabetes; fasting glucose is specific but not sensitive. Interventions in people classified through screening as having pre-diabetes have some efficacy in preventing or delaying onset of type 2 diabetes in trial populations. As screening is inaccurate, many people will receives an incorrect diagnosis and be referred on for interventions while others will be falsely reassured and not offered the intervention. These findings suggest that "screen and treat" policies alone are unlikely to have substantial impact on the worsening epidemic of type 2 diabetes. PROSPERO (No CRD42016042920).


Magnetic reconnection in Saturn's magnetotail: A comprehensive magnetic field survey.
Reconnection within planetary magnetotails is responsible for locally energizing particles and changing the magnetic topology. Its role in terms of global magnetospheric dynamics can involve changing the mass and flux content of the magnetosphere. We have identified reconnection related events in spacecraft magnetometer data recorded during Cassini's exploration of Saturn's magnetotail. The events are identified from deflections in the north-south component of the magnetic field, significant above a background level. Data were selected to provide full tail coverage, encompassing the dawn and dusk flanks as well as the deepest midnight orbits. Overall 2094 reconnection related events were identified, with an average rate of 5.0 events per day. The majority of events occur in clusters (within 3 h of other events). We examine changes in this rate in terms of local time and latitude coverage, taking seasonal effects into account. The observed reconnection rate peaks postmidnight with more infrequent but steady loss seen on the dusk flank. We estimate the mass loss from the event catalog and find it to be insufficient to balance the input from the moon Enceladus. Several reasons for this discrepancy are discussed. The reconnection X line location appears to be highly variable, though a statistical separation between events tailward and planetward of the X line is observed at a radial distance of between 20 and 30RS downtail. The small sample size at dawn prevents comprehensive statistical comparison with the dusk flank observations in terms of flux closure.


Graph analysis of fragmented long-read bacterial genome assemblies.
Long-read genome assembly tools are expected to reconstruct bacterial genomes nearly perfectly; however, they still produce fragmented assemblies in some cases. It would be beneficial to understand whether these cases are intrinsically impossible to resolve, or if assemblers are at fault, implying that genomes could be refined or even finished with little to no additional experimental cost. We propose a set of computational techniques to assist inspection of fragmented bacterial genome assemblies, through careful analysis of assembly graphs. By finding paths of overlapping raw reads between pairs of contigs, we recover potential short-range connections between contigs that were lost during the assembly process. We show that our procedure recovers 45% of missing contig adjacencies in fragmented Canu assemblies, on samples from the NCTC bacterial sequencing project. We also observe that a simple procedure based on enumerating weighted Hamiltonian cycles can suggest likely contig orderings. In our tests, the correct contig order is ranked first in half of the cases and within the top-three predictions in nearly all evaluated cases, providing a direction for finishing fragmented long-read assemblies. https://gitlab.inria.fr/pmarijon/knot . Supplementary data are available at Bioinformatics online.


AF10 (MLLT10) prevents somatic cell reprogramming through regulation of DOT1L-mediated H3K79 methylation.
The histone H3 lysine 79 (H3K79) methyltransferase DOT1L is a key chromatin-based barrier to somatic cell reprogramming. However, the mechanisms by which DOT1L safeguards cell identity and somatic-specific transcriptional programs remain unknown. We employed a proteomic approach using proximity-based labeling to identify DOT1L-interacting proteins and investigated their effects on reprogramming. Among DOT1L interactors, suppression of AF10 (MLLT10) via RNA interference or CRISPR/Cas9, significantly increases reprogramming efficiency. In somatic cells and induced pluripotent stem cells (iPSCs) higher order H3K79 methylation is dependent on AF10 expression. In AF10 knock-out cells, re-expression wild-type AF10, but not a DOT1L binding-impaired mutant, rescues overall H3K79 methylation and reduces reprogramming efficiency. Transcriptomic analyses during reprogramming show that AF10 suppression results in downregulation of fibroblast-specific genes and accelerates the activation of pluripotency-associated genes. Our findings establish AF10 as a novel barrier to reprogramming by regulating H3K79 methylation and thereby sheds light on the mechanism by which cell identity is maintained in somatic cells.


An implementation framework for wastewater treatment models requiring a minimum programming expertise.
Mathematical modelling in environmental biotechnology has been a traditionally difficult resource to access for researchers and students without programming expertise. The great degree of flexibility required from model implementation platforms to be suitable for research applications restricts their use to programming expert users. More user friendly software packages however do not normally incorporate the necessary flexibility for most research applications. This work presents a methodology based on Excel and Matlab-Simulink for both flexible and accessible implementation of mathematical models by researchers with and without programming expertise. The models are almost fully defined in an Excel file in which the names and values of the state variables and parameters are easily created. This information is automatically processed in Matlab to create the model structure and almost immediate model simulation, after only a minimum Matlab code definition, is possible. The framework proposed also provides programming expert researchers with a highly flexible and modifiable platform on which to base more complex model implementations. The method takes advantage of structural generalities in most mathematical models of environmental bioprocesses while enabling the integration of advanced elements (e.g. heuristic functions, correlations). The methodology has already been successfully used in a number of research studies.


Malignancies, prothrombotic mutations, and the risk of venous thrombosis.
Venous thrombosis is a common complication in patients with cancer, leading to additional morbidity and compromising quality of life. To identify individuals with cancer with an increased thrombotic risk, evaluating different tumor sites, the presence of distant metastases, and carrier status of prothrombotic mutations. A large population-based, case-control (Multiple Environmental and Genetic Assessment [MEGA] of risk factors for venous thrombosis) study of 3220 consecutive patients aged 18 to 70 years, with a first deep venous thrombosis of the leg or pulmonary embolism, between March 1, 1999, and May 31, 2002, at 6 anticoagulation clinics in the Netherlands, and separate 2131 control participants (partners of the patients) reported via a questionnaire on acquired risk factors for venous thrombosis. Three months after discontinuation of the anticoagulant therapy, all patients and controls were interviewed, a blood sample was taken, and DNA was isolated to ascertain the factor V Leiden and prothrombin 20210A mutations. Risk of venous thrombosis. The overall risk of venous thrombosis was increased 7-fold in patients with a malignancy (odds ratio [OR], 6.7; 95% confidence interval [CI], 5.2-8.6) vs persons without malignancy. Patients with hematological malignancies had the highest risk of venous thrombosis, adjusted for age and sex (adjusted OR, 28.0; 95% CI, 4.0-199.7), followed by lung cancer and gastrointestinal cancer. The risk of venous thrombosis was highest in the first few months after the diagnosis of malignancy (adjusted OR, 53.5; 95% CI, 8.6-334.3). Patients with cancer with distant metastases had a higher risk vs patients without distant metastases (adjusted OR, 19.8; 95% CI, 2.6-149.1). Carriers of the factor V Leiden mutation who also had cancer had a 12-fold increased risk vs individuals without cancer and factor V Leiden (adjusted OR, 12.1; 95% CI, 1.6-88.1). Similar results were indirectly calculated for the prothrombin 20210A mutation in patients with cancer. Patients with cancer have a highly increased risk of venous thrombosis especially in the first few months after diagnosis and in the presence of distant metastases. Carriers of the factor V Leiden and prothrombin 20210A mutations appear to have an even higher risk.


Tests of the accuracy and speed of categorizing foods into child vs professional categories using two methods of browsing with children.
This research tested whether children could categorize foods more accurately and speedily when presented with child-generated rather than professionally generated food categories, and whether a graphically appealing browse procedure similar to the Apple iTunes (Cupertino, CA) "cover flow" graphical user interface accomplished this better than the more common tree-view structure. In Fall 2008, 104 multiethnic children ages 8 to 13 were recruited at the Baylor College of Medicine (Houston, TX) and randomly assigned to two browse procedures: cover flow (collages of foods in a category) or tree view (food categories in a list). Within each browse condition children categorized the same randomly ordered 26 diverse foods to both child and professionally organized categories (with method randomly sequenced per child). Acceptance of categorization was determined by registered dietitians. Speed of categorization was recorded by the computer. Differences between methods were determined by repeated measures analysis of variance. Younger children (8 to 9 years old) tended to have lower acceptance and longer speeds of categorization. The quickest categorization was obtained with child categories in a tree structure. Computerized dietary reporting by children can use child-generated food categories and tree structures to organize foods for browsing in a hierarchically organized structure to enhance speed of categorization, but not accuracy. A computerized recall may not be appropriate for children 9 years of age or younger.


Development of a Stable Lung Microbiome in Healthy Neonatal Mice.
The lower respiratory tract has been previously considered sterile in a healthy state, but advances in culture-independent techniques for microbial identification and characterization have revealed that the lung harbors a diverse microbiome. Although research on the lung microbiome is increasing and important questions were already addressed, longitudinal studies aiming to describe developmental stages of the microbial communities from the early neonatal period to adulthood are lacking. Thus, little is known about the early-life development of the lung microbiome and the impact of external factors during these stages. In this study, we applied a barcoding approach based on high-throughput sequencing of 16S ribosomal RNA gene amplicon libraries to determine age-dependent differences in the bacterial fraction of the murine lung microbiome and to assess potential influences of differing "environmental microbiomes" (simulated by the application of used litter material to the cages). We could clearly show that the diversity of the bacterial community harbored in the murine lung increases with age. Interestingly, bacteria belonging to the genera Delftia and Rhodococcus formed an age-independent core microbiome. The addition of the used litter material influenced the lung microbiota of young mice but did not significantly alter the community composition of adult animals. Our findings elucidate the dynamic nature of the early-life lung microbiota and its stabilization with age. Further, this study indicates that even slight environmental changes modulate the bacterial community composition of the lung microbiome in early life, whereas the lung microbes of adults demonstrate higher resilience towards environmental variations.


Hot compaction of poly(methyl methacrylate) composites based on fiber shrinkage results.
Uniaxial self-reinforced composite poly(methyl methacrylate) (SRC-PMMA) is being investigated as a pre-coat material for the femoral component of total hip replacements. Hot compaction of self-reinforced composites is largely an empirical process which varies the processing parameters of time, temperature and pressure until the desired properties are obtained. Previous work has shown that PMMA fibers have unique thermal relaxation properties dependent upon the retained molecular orientation in them. This work processed composites at times and temperatures that span the relaxation process for a single fiber. It was found that molecular orientation, as measured by birefringence, was lost in composites processed at times greater than relaxation times for single fibers. Flexural properties were also found to vary with processing conditions, with the highest values of 165 +/- 15 MPa and 168 +/- 3 MPa found at high and low processing times, respectively. These are significantly stronger than unreinforced PMMA which has a flexural strength of 127 +/- 14 MPa. It is hypothesized that diffusion between fibers occurs much more quickly than the loss of molecular orientation and it was seen that SRC-PMMA processing conditions can be predicted from the relaxation times and temperatures from single fibers.


A Robust and General Approach to Quantitatively Conjugate Enzymes to Plasmonic Nanoparticles.
Bioconjugates of plasmonic nanoparticles have received considerable attention due to their potential biomedical applications. Successful bioconjugation requires control over the number and activity of the conjugated proteins and the colloidal stability of the particles. In practice, this requires reoptimization of the conjugation protocol for each combination of protein and nanoparticle. Here, we report a robust and general protocol that allows for the conjugation of a range of proteins to different types of nanoparticles using very short polyethylene-glycol(PEG) linkers, while simultaneously preserving protein activity and colloidal stability. The use of short linkers ensures that the protein is located close to the particle surface, where the refractive index sensitivity and near-field enhancement are maximal. We demonstrate that the use of a Tween20 containing stabilizing buffer is critical in maintaining colloidal stability and protein function throughout the protocol. We obtain quantitative control over the average number of enzymes per particle by either varying the number of functional groups on the particle or the enzyme concentration during incubation. This new route of preparing quantitative protein-nanoparticle bioconjugates paves the way to develop rational and quantitative strategies to functionalize nanoparticles for applications in sensing, medical diagnostics, and drug delivery.


Estimated historical and current nitrogen balances for Illinois.
The Midwest has large riverine exports of nitrogen (N), with the largest flux per unit area to the Mississippi River system coming from Iowa and Illinois. We used historic and current data to estimate N inputs, outputs, and transformations for Illinois where human activity (principally agriculture and associated landscape drainage) have had a dominant impact. Presently, approximately 800,000 Mg of N is added each year as fertilizer and another 420,000 Mg is biologically fixed, primarily by soybean (Glycine max L. Merr.). These annual inputs are greater than exports in grain, which results in surplus N throughout the landscape. Rivers within the state export approximately 50% of this surplus N, mostly as nitrate, and the remainder appears to be denitrified or temporarily incorporated into the soil organic matter pool. The magnitude of N losses for 1880, 1910, 1950, and 1990 are compared. Initial cultivation of the prairies released large quantities of N (approximately 500,000 Mg N year(-1)), and resulted in riverine N transport during the late 19th century that appears to have been on the same order of magnitude as contemporary N losses. Riverine flux was estimated to have been at a minimum in about 1950, due to diminished net mineralization and low fertilizer inputs. Residual fertilizer N from corn (Zea mays L.), biological N fixed by soybean, short-circuiting of soil water through artificial drainage, and decreased cropping-system diversity appear to be the primary sources for current N export.


Notch3 establishes brain vascular integrity by regulating pericyte number.
Brain pericytes are important regulators of brain vascular integrity, permeability and blood flow. Deficiencies of brain pericytes are associated with neonatal intracranial hemorrhage in human fetuses, as well as stroke and neurodegeneration in adults. Despite the important functions of brain pericytes, the mechanisms underlying their development are not well understood and little is known about how pericyte density is regulated across the brain. The Notch signaling pathway has been implicated in pericyte development, but its exact roles remain ill defined. Here, we report an investigation of the Notch3 receptor using zebrafish as a model system. We show that zebrafish brain pericytes express notch3 and that notch3 mutant zebrafish have a deficit of brain pericytes and impaired blood-brain barrier function. Conditional loss- and gain-of-function experiments provide evidence that Notch3 signaling positively regulates brain pericyte proliferation. These findings establish a new role for Notch signaling in brain vascular development whereby Notch3 signaling promotes expansion of the brain pericyte population.


Separate effects of triploidy, parentage and genomic diversity upon feeding behaviour, metabolic efficiency and net energy balance in the Pacific oyster Crassostrea gigas.
Triploid oysters were induced using cytochalasin B upon retention of either the first (meiosis I triploids) or the second (meiosis II triploids) polar body in embryos from a single cohort derived from mixed parentage. Allozyme and microsatellite assays enabled the confirmation of both parentage and triploidy status in each oyster. Comparison of meiosis I triploids, meiosis II triploids and diploid siblings established that improved physiological performance in triploids was associated with increased allelic variation, rather than with the quantitative dosage effects of ploidy status. An unidentified maternal influence also interacted with genotype. Among full sibs, allelic variation measured as multi-locus enzyme heterozygosity accounted for up to 42% of the variance in physiological performance; significant positive influences were identified upon feeding rate, absorption efficiency, net energy balance and growth efficiency (= net energy balance divided by energy absorbed). Whilst allelic variation was greater in both meiosis I and meiosis II triploids than in diploid siblings, both allelic variation and net energy balance were highest in triploids induced at meiosis I. This suggests that it may be preferable to induce triploidy by blocking meiosis I, rather than meiosis II as has traditionally been undertaken during commercial breeding programmes.


Identification of semaphorin 4B as a negative regulator of basophil-mediated immune responses.
Basophils are strong mediators of Th2 responses during helminthic infections. Recently, basophils were shown to function as APCs and promote both Th2 skewing and humoral memory responses. However, the mechanisms that regulate basophils are still unclear. In this article, we show that a class IV semaphorin, Sema4B, negatively regulates basophil functions through T cell-basophil contacts. In a screen to identify semaphorins that function in the immune system, we determined that Sema4B is expressed in T and B cells. Interestingly, Sema4B(-/-) mice had considerably increased serum IgE levels despite normal lymphocyte and dendritic cell functions. Recombinant Sema4B significantly inhibited IL-4 and IL-6 production from basophils in response to various stimuli, including IL-3, papain, and FcεRI cross-linking. In addition, T cell-derived Sema4B, which accumulated at contact sites between basophils and CD4(+) T cells, suppressed basophil-mediated Th2 skewing, suggesting that Sema4B regulates basophil responses through cognate cell-cell contacts. Furthermore, Sema4B(-/-) mice had enhanced basophil-mediated memory IgE production, which was abolished by treating with an anti-FcεRIα Ab. Collectively, these results indicate that Sema4B negatively regulates basophil-mediated Th2 and humoral memory responses.


A meta-analysis of clinical studies conducted during the West Africa Ebola virus disease outbreak confirms the need for randomized control groups.
Recent Ebola virus disease outbreaks affirm the dire need for treatments with proven efficacy. Randomized controlled clinical trials remain the gold standard but, during disease outbreaks, may be difficult to conduct due to ethical concerns and challenging field conditions. In the absence of a randomized control group, statistical modeling to create a control group could be a possibility. Such a model-based reference control would only be credible if it had the same mortality risk as that of the experimental group in the absence of treatment. One way to test this counterfactual assumption is to evaluate whether reasonable similarity exists across nonrandomized control groups from different clinical studies, which might suggest that a future control group would be similarly homogeneous. We evaluated similarity across six clinical studies conducted during the 2013-2016 West Africa outbreak of Ebola virus disease. These studies evaluated favipiravir, the biologic ZMapp, the antimalarial drug amodiaquine, or administration of convalescent plasma or convalescent whole blood. We compared the nonrandomized control groups of these six studies comprising 1147 individuals infected with Ebola virus. We found considerable heterogeneity, which did not disappear after statistical modeling to adjust for prognostic variables. Mortality risk varied widely (31 to 66%) across the nonrandomized control arms of these six studies. Models adjusting for baseline covariates (age, sex, and cycle threshold, a proxy for viral load) failed to sufficiently recalibrate these studies and showed that heterogeneity remained. Our findings highlight concerns about making invalid conclusions when comparing nonrandomized control groups to cohorts receiving experimental treatments.


Quantitative localization of NMDAR1 receptor subunit immunoreactivity in inferotemporal and prefrontal association cortices of monkey and human.
The cellular and synaptic localization of immunoreactivity for the N-methyl-D-aspartate (NMDA) receptor subunit, NMDAR1, was investigated in inferotemporal and prefrontal association neocortices of monkeys and humans. In all monkey association areas examined, the laminar distribution patterns of NMDAR1 immunoreactivity were similar, and characterized by predominant pyramidal-like neuronal labeling in layers II, III, V and VI and a dense neuropil labeling consisting of intensely stained puncta and fine-caliber processes present throughout layers I-III, and V-VI. Layer IV, in contrast, contained only very lightly immunostained neurons which mostly lacked extensive dendritic staining. The laminar distribution of NMDAR1 immunolabeling in human association cortex was similar to that observed in monkeys. Electron microscopy of monkey areas 46 and TE1 confirmed that intensely immunoreactive asymmetrical postsynaptic densities were present throughout all cell-dense layers of prefrontal and inferotemporal association cortex. Quantitative analyses of the laminar proportions of immunoreactive synapses demonstrated that in both areas examined, the percentages of immunolabeled synapses were mostly similar across superficial layers, layer IV and infragranular layers. Finally, quantitative double-labeling immunofluorescence for non-NMDA receptor subunits or calcium-binding proteins demonstrated that virtually all GluR2/3 or GluR5/6/7-immunoreactive neurons were also labeled for NMDAR1, while regionally-specific subsets of parvalbumin-, calbindin- and calretinin-immunoreactive neurons were co-labeled. These data indicate that in primate association cortex, NMDA receptors are heterogeneously distributed to subsets of functionally distinct types of neurons and subsets of excitatory synapses, suggesting a critical and highly specific role in mediating the activity of excitatory connectivity which converges on cortical association areas.


ABHD10 is an S-depalmitoylase affecting redox homeostasis through peroxiredoxin-5.
S-Palmitoylation is a reversible lipid post-translational modification that has been observed on mitochondrial proteins, but both the regulation and functional consequences of mitochondrial S-palmitoylation are poorly understood. Here, we show that perturbing the 'erasers' of S-palmitoylation, acyl protein thioesterases (APTs), with either pan-active inhibitors or a mitochondrial-targeted APT inhibitor, diminishes the antioxidant buffering capacity of mitochondria. Surprisingly, this effect was not mediated by the only known mitochondrial APT, but rather by a resident mitochondrial protein with no known endogenous function, ABHD10. We show that ABHD10 is a member of the APT family of regulatory proteins and identify peroxiredoxin-5 (PRDX5), a key antioxidant protein, as a target of ABHD10 S-depalmitoylase activity. We then find that ABHD10 regulates the S-palmitoylation status of the nucleophilic active site residue of PRDX5, providing a direct mechanistic connection between ABHD10-mediated S-depalmitoylation of PRDX5 and its antioxidant capacity.


Clotrimazole ameliorates intestinal inflammation and abnormal angiogenesis by inhibiting interleukin-8 expression through a nuclear factor-kappaB-dependent manner.
Increased interleukin (IL)-8 plays an important role not only in activation and recruitment of neutrophils but also in inducing exaggerated angiogenesis at the inflamed site. In the present study, we investigated the fact that clotrimazole (CLT) inhibits intestinal inflammation, and the inhibitory action is mediated through suppression of IL-8 expression. In the trinitrobenzene sulfonic acid (TNBS)-induced rat colitis model, CLT dose-dependently protected from the TNBS-induced weight loss, colon ulceration, and myeloperoxidase activity increase. In the lesion site, CLT also suppressed the TNBS-induced angiogenesis, IL-8 expression, and nuclear factor (NF)-kappaB activation. In a cellular model of colitis using tumor necrosis factor (TNF)-alpha-stimulated HT29 colon epithelial cells, treatment with CLT significantly suppressed TNF-alpha-mediated IL-8 induction and NF-kappaB transcriptional activity revealed by a luciferase reporter gene assay. Furthermore, cotreatment with CLT and pyrrolidine dithiocarbamate, a NF-kappaB inhibitor, synergistically reduced the NF-kappaB transcriptional activity as well as IL-8 expression. In an in vitro angiogenesis assay, CLT suppressed IL-8-induced proliferation, tube formation, and invasion of human umbilical vein endothelial cells. The in vivo angiogenesis assay using chick chorioallantoic membrane also showed that CLT significantly inhibited the IL-8-induced formation of new blood vessels. Taken together, these results suggest that CLT may prevent the progression of intestinal inflammation by not only down-regulating IL-8 expression but also inhibiting the action of IL-8 in both colon epithelial and vascular endothelial cells during pathogenesis of intestinal inflammation.


Stem cell therapy for inflammatory bowel disease.
Inflammatory bowel disease (IBD) could be curable by "immune rest" and correction of the genetic predisposition inherent in allogeneic hematopoietic stem cell transplantation. However, balancing risks against benefits remains challenging. The application of mesenchymal stem cells (MSCs) serving as a site-regulated "drugstore" is a recent concept, which suggests the possibility of an alternative treatment for many intractable diseases such as IBD. Depending on the required function of MSC, such as a cell provider, immune moderator, and/or trophic resource, MSC therapy should be optimized to maximize its therapeutic benefit. Therapeutic effects do not always require full engraftment of MSCs. Therefore, optimization of pleiotropic gut trophic factors produced by MSCs, which favoring not only regulating immune responses but also promoting tissue repair, must directly enhance new drug discoveries for treatment of IBD. Stem cell biology holds great promise for a new era of cell-based therapy, sparking considerable interest among scientists, clinicians, and patients. However, the translational arm of stem cell science remains in a relatively primitive state. Although several clinical studies using MSCs have been initiated, early results suggest several inherent problems. In each study, optimization of MSC therapy appears to be the most urgent problem, and can be resolved only by scientifically unveiling the mechanisms of therapeutic action. In the present review, the authors outline how such information would facilitate the critical steps in the paradigm shift from basic research on stem cell biology to clinical practice of regenerative medicine for conquering IBD in the near future.


Novel tarantula toxins for subtypes of voltage-dependent potassium channels in the Kv2 and Kv4 subfamilies.
Three novel peptides with the ability to inhibit voltage-dependent potassium channels in the shab (Kv2) and shal (Kv4) subfamilies were identified from the venom of the African tarantulas Stromatopelma calceata (ScTx1) and Heteroscodra maculata (HmTx1, HmTx2). The three toxins are 34- to 38-amino acid peptides that belong to the structural family of inhibitor cystine knot spider peptides reticulated by three disulfide bridges. Electrophysiological recordings in COS cells show that these toxins act as gating modifier of voltage-dependent K+ channels. ScTx1 is the first high-affinity inhibitor of the Kv2.2 channel subtype (IC50, 21.4 nM) to be described. ScTx1 also inhibits the Kv2.1 channels, with an IC50 of 12.7 nM, and Kv2.1/Kv9.3 heteromultimers that have been proposed to be involved in O2 sensing in pulmonary artery myocytes. In addition, it is the most effective inhibitor of Kv4.2 channels described thus far, with an IC50 of 1.2 nM. HmTx toxins share sequence similarities with both the potassium channel blocker toxins (HmTx1) and the calcium channel blocker toxin omega-GsTx SIA (HmTx2). They inhibit potassium current associated with Kv2 subtypes in the 100 to 300 nM concentration range. HmTx2 seems to be a specific inhibitor of Kv2 channels, whereas HmTx1 also inhibits Kv4 channels, including Kv4.1, with the same potency. HmTx1 is the first described peptide effector of the Kv4.1 subtype. Those novel toxins are new tools for the investigation of the physiological role of the different potassium channel subunits in cellular physiology.


Infant-mother attachment security, contextual risk, and early development: a moderational analysis.
In light of evidence that the effects of attachment security on subsequent development may be contingent on the social context in which the child continues to develop, we examined the effect of attachment security at age 15 months, cumulative contextual risk from 1 to 36 months, and the interaction of attachment and cumulative risk to predict socioemotional and cognitive linguistic functioning at age 3 years, using data from the National Institute of Child Health and Human Development Study of Early Child Care. Results indicated that early attachment predicts both socioemotional development and language skills, but not cognitive functioning as indexed by a measure of school readiness, and that the effect of attachment on socioemotional development and expressive language varied as a function of social-contextual risk. Insecure-avoidant infants proved most vulnerable to contextual risk, not children classified as secure or insecure more generally, although in one instance security did prove protective with respect to the adverse effects of cumulative contextual risk. Findings are discussed in terms of risk and resilience and in light of the probabilistic nature of the relation between early attachment and later development.


Structure and electronic spectra of DNA mini-hairpins with G(n):C(n) stems.
The solution structure of a synthetic DNA mini-hairpin possessing a stilbenediether linker and three G:C base pairs has been obtained using (1)H NMR spectral data and constrained torsion angle molecular dynamics. Notable features of this structure include a compact hairpin loop having a short stilbene-guanine plane-to-plane distance and approximate B-DNA geometry for the three base pairs. Comparison of the electronic spectra of mini-hairpins having one-to-four G:C base pairs and stilbenediether or hexamethyleneglycol linkers reveals the presence of features in the UV and CD spectra of the stilbene-linked hairpins that are not observed for the ethyleneglycol-linked hairpins. Investigation of the electronic structure of a stilbene-linked hairpin having a single G:C base pair by means of time-dependent density functional theory shows that the highest occupied molecular orbital, but not the lowest unoccupied molecular orbital, is delocalized over the stilbene and adjacent guanine. The calculated UV and CD spectra are highly dependent upon hairpin conformation, but reproduce the major features of the experimental spectra. These results illustrate the utility of an integrated experimental and theoretical approach to understanding the complex electronic spectra of pi-stacked chromophores.


Breast feeding, but not use of oral contraceptives, is associated with a reduced risk of rheumatoid arthritis.
To determine whether breast feeding or the use of oral contraceptives (OCs) affects the future risk of rheumatoid arthritis (RA) in a community-based prospective cohort. A community-based health survey (18 326 women) was linked to regional and national registers, and incident cases of RA were identified. All women with a diagnosis of RA after inclusion in the health survey (n = 136) and four female controls for every case, who were alive and free from RA when the index person was given a diagnosis of RA, were included in a case-control study. Data on lifestyle factors at baseline were derived from a self-administered questionnaire. Potential predictors were examined in logistic regression models. 136 women with incident RA were compared with 544 age-matched controls. A longer history of breast feeding was associated with a reduced risk of RA (OR 0.46 (95% CI 0.24 to 0.91) for women who had breast fed for >/=13 months and OR 0.74 (95% CI 0.45 to 1.20) for those who had breast fed for 1-12 months, compared with those who had never breast fed). The protective effect of longer breast feeding remained significant after adjustment for smoking and level of education in multivariate models, and point estimates were protective also when the analyses were restricted to parous women. Neither parity nor OC use had any significant effect on the risk of RA. In this study, long-term breast feeding, but not OC use, was associated with a significant reduction in the risk of RA.


Testing a model of facilitated reflection on network feedback: a mixed method study on integration of rural mental healthcare services for older people.
To test a management model of facilitated reflection on network feedback as a means to engage services in problem solving the delivery of integrated primary mental healthcare to older people. Participatory mixed methods case study evaluating the impact of a network management model using organisational network feedback (through social network analysis, key informant interviews and policy review). A model of facilitated network reflection using network theory and methods. A rural community in South Australia. 32 staff from 24 services and 12 senior service managers from mental health, primary care and social care services. Health and social care organisations identified that they operated in clustered self-managed networks within sectors, with no overarching purposive older people's mental healthcare network. The model of facilitated reflection revealed service goal and role conflicts. These discussions helped local services to identify as a network, and begin the problem-solving communication and referral links. A Governance Group assisted this process. Barriers to integrated servicing through a network included service funding tied to performance of direct care tasks and the lack of a clear lead network administration organisation. A model of facilitated reflection helped organisations to identify as a network, but revealed sensitivity about organisational roles and goals, which demonstrated that conflict should be expected. Networked servicing needed a neutral network administration organisation with cross-sectoral credibility, a mandate and the resources to monitor the network, to deal with conflict, negotiate commitment among the service managers, and provide opportunities for different sectors to meet and problem solve. This requires consistency and sustained intersectoral policies that include strategies and funding to facilitate and maintain health and social care networks in rural communities.


Transoral resection of axial lesions augmented by intraoperative magnetic resonance imaging. Report of three cases.
Transoral decompression of the cervicomedullary junction may be compromised by a narrow corridor in which surgery is performed, and thus the adequacy of surgical decompression/resection may be difficult to determine. This is problematic as the presence of spinal instrumentation may obscure the accuracy of postoperative radiological assessment, or the patient may require reoperation. The authors describe three patients in whom high-field intraoperative magnetic resonance (MR) images were acquired at various stages during the transoral resection of C-2 disease that had caused craniocervical junction compression. All three patients harbored different lesions involving the cervicomedullary junction: one each of plasmacytoma and metastatic breast carcinoma involving the odontoid process and C-2 vertebral body, and basilar invagination with a Chiari I malformation. All patients presented with progressive myelopathy. Surgical planning MR imaging studies performed after the induction of anesthesia demonstrated the lesion and its relationship to the planned surgical corridor. Transoral exposure was achieved through placement of a Crockard retractor system. In one case the soft palate was divided. Interdissection MR imaging revealed that adequate decompression had been achieved in all cases. The two patients with carcinoma required placement of posterior instrumentation for stabilization. Planned suboccipital decompression and placement of instrumentation were averted in the third case as the intraoperative MR images demonstrated that excellent decompression had been achieved. Intraoperatively acquired MR images were instrumental in determining the adequacy of the decompressive surgery. In one of the three cases, examination of the images led the authors to change the planned surgical procedure. Importantly, the acquisition of intraoperative MR images did not adversely affect operating time or neurosurgical techniques, including instrumentation requirements.


Central and peripheral fatigue during passive and exercise-induced hyperthermia.
Hyperthermia was induced during prolonged exercise (ExH) and passive heating (PaH) to isolate the influence of exercise on neuromuscular function during a maximal voluntary isometric contraction (MVC) of the quadriceps under heat stress. The influence of cardiovascular strain in limiting endurance performance in the heat was also examined. On separate days, eight males cycled to exhaustion at 60% maximal oxygen uptake or were immersed in a water bath (∼41°C) until rectal temperature (Tre) increased to 39.5°C. The ExH and PaH interventions were performed in ambient conditions of 38°C and 60% relative humidity with Tre reaching 39.8°C during exercise. Before (control) and after each intervention, voluntary activation and force production capacity were evaluated by superimposing an electrically stimulated tetanus during a 45-s MVC. Force production decreased immediately after PaH and ExH compared with control, with the magnitude of decline being more pronounced after ExH (P < 0.01). Mean voluntary activation was also significantly depressed after both interventions (P < 0.01 vs control). However, the extent of decline in voluntary activation was maintained at ∼90% during both PaH and ExH MVC. This decline accounted for 41.5% (PaH) and 33.1% (ExH) of the decrease in force production. In addition, exhaustion coincided with a marked increase in HR (∼96% of maximum) and a decline in stroke volume (25%) and mean arterial pressure (10%) (P < 0.05). The loss of force production capacity during hyperthermia originated from central and peripheral fatigue factors, with the combination of heat stress and previous contractile activity exacerbating the rate of decline. Thus, the observed significant rise in thermal strain in ExH and PaH impaired neuromuscular function and was associated with an exercise performance limiting increase in cardiovascular strain.


Intensive conventional chemotherapy (ACVBP regimen) compared with standard CHOP for poor-prognosis aggressive non-Hodgkin lymphoma.
We conducted a randomized trial to compare the intensive conventional chemotherapy regimen ACVBP (doxorubicin, cyclophosphamide, vindesine, bleomycin, prednisone) with standard CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) in previously untreated patients with poor-risk aggressive lymphoma. Patients aged 61 to 69 years who had aggressive non-Hodgkin lymphoma with at least one prognostic factor of the age-adjusted international prognostic index (IPI) were included. ACVBP consisted of an induction phase of intensified chemotherapy and central nervous system (CNS) prophylaxis followed by a sequential consolidation phase. Of the 708 patients registered for the study, 635 were eligible. The rate of complete response was 58% in the ACVBP group and 56% in the CHOP group (P =.5). Treatment-related death occurred in 13% of the ACVBP group and 7% of the CHOP group (P =.014). At 5 years, the event-free survival was 39% in the ACVBP group and 29% in the CHOP group (P =.005). The overall survival was significantly longer for patients treated with ACVBP, at 5 years it was 46% compared with 38% for patients treated with CHOP (P =.036). CNS progressions or relapses were more frequent in the CHOP group (P =.004). Despite higher toxicity, the ACVBP regimen, used as first-line treatment for patients with poor-risk aggressive lymphoma, is superior to standard CHOP with regard to both event-free survival and overall survival.


Diagnosis and treatment for ocular tuberculosis among uveitis specialists: the international perspective.
To assess the approach of international specialists, who primarily practice in tuberculosis-endemic areas, to ocular tuberculosis (TB). International experts from India, Brazil, Taiwan, and more than 10 other countries were surveyed using two clinical cases and general questions. A total of 244 experts were sent a survey about the treatment and diagnosis of ocular tuberculosis; 65 responded (27%), of whom 34 were affiliated with practices in India, while 31 primarily practice at international sites outside of India and North America. The data from this survey were compared with the results of a similar survey sent to members of the American Uveitis Society. The survey provided normative data on how physicians evaluate patients with uveitis as well as opinions about ocular TB. Responses varied widely on topics such as tests to include in the workup of undifferentiated uveitis, initial therapy, and duration of treatment. Physicians from developing countries relied more on chest CT scans and tuberculin skin testing (TST) than their counterparts in developed countries. The approach to diagnosis and management of TB is heterogeneous worldwide. However, there are substantial differences in the clinical approach to uveitis depending on the clinician's location of practice.


Cannabis Use in Children With Pantothenate Kinase-Associated Neurodegeneration.
Pantothenate kinase-associated neurodegeneration is characterized by severe, progressive dystonia. This study aims to describe the reported usage of cannabis products among children with pantothenate kinase-associated neurodegeneration. A cross-sectional, 37-item survey was distributed in April 2019 to the families of 44 children who participate in a clinical registry of individuals with pantothenate kinase-associated neurodegeneration. We received 18 responses (40.9% response rate). Children were a mean of 11.0 (SD 4.3) years old. The 15 respondents with dystonia or spasticity were on a median of 2 tone medications (range 0-9). Seven children had ever used cannabis (38.9%). The most common source of information about cannabis was other parents. Children who had ever used cannabis were on more tone medications, were more likely to have used opiates, were less likely to be able to roll, and less likely to sit comfortably, than children who had never used cannabis. Four children reported moderate or significant improvement in dystonia with cannabis. Other areas reported to be moderate or significantly improved were pain (n = 3), sleep (n = 4), anxiety (n = 3), and behavior (n = 2). Adverse effects included sadness (n = 1), agitation/behavior change (n = 1), and tiredness (n = 1). Cannabis use was commonly reported among children with pantothenate kinase-associated neurodegeneration whose parents responded to a survey, particularly when many other dystonia treatments had been tried. Physicians should be aware that parents may treat their child with severe, painful dystonia with cannabis. Placebo-controlled studies of products containing cannabidiol and 9-tetrahydrocannabinol are needed for pediatric tone disorders.


Tasimelteon for the treatment of non-24-hour sleep-wake disorder.
Tasimelteon (Hetlioz®), a melatonin receptor agonist, is the first, and, at the time of the publication, the only drug to be approved by the U.S. Food and Drug Administration (FDA) for the treatment of non-24-hour sleep-wake disorder (non-24). This circadian rhythm disorder occurs most commonly in blind individuals without light perception, and it results from their inability to entrain to the 24-hour photoperiod, although the indication does not specify a particular patient population. Non-24 is characterized by a persistent cycle of nighttime insomnia and daytime sleepiness, alternating with asymptomatic periods depending on an individual's degree of circadian rhythm synchronization with the photoperiod at any particular time. Phase II clinical trials in healthy individuals confirmed the circadian phase-shifting potential of tasimelteon. Phase III trials in totally blind subjects diagnosed with non-24 demonstrated the efficacy of tasimelteon in reducing both nighttime wakefulness and daytime napping. Physiologic monitoring revealed that tasimelteon resulted in a higher proportion of individuals becoming entrained to the 24-hour cycle compared with placebo. Safety assessments indicated that tasimelteon is well tolerated, with the most common adverse events being headache, alanine aminotransferase elevation, nightmares or unusual dreams, and upper respiratory or urinary tract infections. Tasimelteon is available as a capsule in a single 20-mg dose and it must be obtained through Vanda Pharmaceutical's HetliozSolutions program with dispensing through a specialty pharmacy. Safety studies in blind individuals diagnosed with non-24 are ongoing and a future clinical trial with Smith-Magenis syndrome patients is planned.


Routine determination of ice thickness for cryo-EM grids.
Recent advances in instrumentation and automation have made cryo-EM a popular method for producing near-atomic resolution structures of a variety of proteins and complexes. Sample preparation is still a limiting factor in collecting high quality data. Thickness of the vitreous ice in which the particles are embedded is one of the many variables that need to be optimized for collection of the highest quality data. Here we present two methods, using either an energy filter or scattering outside the objective aperture, to measure ice thickness for potentially every image collected. Unlike geometrical or tomographic methods, these can be implemented directly in the single particle collection workflow without interrupting or significantly slowing down data collection. We describe the methods as implemented into the Leginon/Appion data collection workflow, along with some examples from test cases. Routine monitoring of ice thickness should prove helpful for optimizing sample preparation, data collection, and data processing.


Disialo-trisialo bridging of transferrin is due to increased branching and fucosylation of the carbohydrate moiety.
Carbohydrate deficient transferrin (CDT) is used for detection of alcohol abuse and follow-up. High performance liquid chromatography (HPLC) of transferrin glycoforms is highly specific for identification of alcohol abuse, but unresolved disialo- and trisialotransferrin glycoforms sometimes makes interpretation difficult. The cause of this phenomenon is unknown, cannot be explained by genetic variants of transferrin, but seems to be associated with liver disease. Nineteen serum samples showing di-tri bridging when analyzed by HPLC were collected. Transferrin was purified by affinity chromatography, and N-linked oligosaccharides were released enzymatically. The N-glycans were further analyzed by high performance anion-exchange chromatography with pulsed amperometric detection and MALDI-TOF mass spectrometry. The HPLC-analysis showed three different types of glycoform patterns. The N-glycans from fifteen samples showed patterns with increased number of triantennary structures containing one or two fucose residues. One sample contained an increased amount of triantennary glycans without fucose. Three samples showed a glycosylation pattern similar to normal transferrin. The di-tri bridging phenomenon was associated with alterations in transferrin glycosylation in the majority of cases. Transferrin contained a higher extent of triantennary and often fucosylated N-linked oligosaccharides. These results may be important in future diagnostic approaches to liver diseases.


Placement of the preparation line and periodontal health--a prospective 2-year clinical study.
The aim of this prospective clinical 2-year study was to determine whether the placement of the proximal margins of crowns within the zone of the biologic width results in periodontal alterations. In 41 patients, 116 prepared teeth as well as 82 unrestored, healthy contralateral teeth were examined. The following indices were determined before as well as 3, 6, 12, and 24 months after preparation: hygiene index, papillary bleeding index (PBI), and probing depth. After preparation, the distance between the restoration margins and the alveolar crest was measured with a modified periodontal probe on both proximal aspects of each tooth. These values were classified into 3 groups: I = < or = 1 mm between crown margin and alveolar crest, II = 1 to 2 mm, and III = > 2 mm. In addition, radiographs were taken directly after cementation of each restoration and after 12 and 24 months. The hygiene index did not significantly vary during the investigation, but PBI increased in all groups. The highest PBI increase was found in group I; in this group, the most significant increase was found between 3 and 6 months after preparation. The mean probing depth only increased in group I during the first 3 months after preparation on the mesial aspects. Only minor changes were found on the distal aspects of the teeth in group I and on all aspects in groups II and III. No alterations of the bone levels were diagnosed on the radiographs. These results indicate that the location of the restoration margins within the zone of the biologic width may impair the periodontal health of restored teeth.


Existence of an endogenous circadian blood pressure rhythm in humans that peaks in the evening.
Blood pressure (BP) usually decreases during nocturnal sleep and increases during daytime activities. Whether the endogenous circadian control system contributes to this daily BP variation has not been determined under appropriately controlled conditions. To determine whether there exists an endogenous circadian rhythm of BP in humans. In 28 normotensive adults (16 men), we assessed BP across 3 complementary, multiday, in-laboratory protocols performed in dim light, throughout which behavioral and environmental influences were controlled and/or uniformly distributed across the circadian cycle via: (1) a 38-hour "constant routine," including continuous wakefulness; (2) a 196-hour "forced desynchrony" with 7 recurring 28-hour sleep/wake cycles; and (3) a 240-hour forced desynchrony with 12 recurring 20-hour sleep/wake cycles. Circadian phases were derived from core body temperature. Each protocol revealed significant circadian rhythms in systolic and diastolic BP, with almost identical rhythm profiles among protocols. The peak-to-trough amplitudes were 3 to 6 mm Hg for systolic BP and 2 to 3 mm Hg for diastolic BP (always P<0.05). All 6 peaks (systolic and diastolic BP in 3 protocols) occurred at a circadian phase corresponding to ≈9:00 pm (ie, the biological evening). Based on substantial phase differences among circadian rhythms of BP and other variables, the rhythm in BP appeared to be unrelated to circadian rhythms in cortisol, catecholamines, cardiac vagal modulation, heart rate, or urine flow. There exists a robust endogenous circadian rhythm in BP. The highest BP occurred at the circadian time corresponding to ≈9:00 pm, suggesting that the endogenous BP rhythm is unlikely to underlie the well-documented morning peak in adverse cardiovascular events.


Physical activity in light of affordances in outdoor environments: qualitative observation studies of 3-5 years olds in kindergarten.
This article examines the characteristic of affordances of different outdoor environments, related to the influences of children's physical activity levels. Qualitative observation studies in a Norwegian kindergarten were conducted of 3- to 5-year-olds into the natural environment and in the kindergarten's outdoor area. An ecological approach was important from both an analytical and theoretical point of view, using concepts from Gibson's (The ecological approach to visual perception. Houghton Mifflin Company, Bosten, 1979) theory of affordances. The concepts of affordances in an environment can explain children's movement behaviour. The findings reveal that situations with high physical activity levels among the children are more often created in natural environments than in the kindergarten's outdoor environment. Natural environments offer potential qualities that are a catalyst for physical activity. The study shows that certain characteristic of the physical outdoor environment are important for children's opportunities and inspiration for physical active play. The findings also show that social possibilities and opportunities, human interactions, in the environment have the greatest influence on the duration and intensity of physically active play. The need for knowledge on physical and social opportunities in outdoor environments, educational practice and the content of outdoor time in kindergartens should be given greater attention.


"A Somali girl is Muslim and does not have premarital sex. Is vaccination really necessary?" A qualitative study into the perceptions of Somali women in the Netherlands about the prevention of cervical cancer.
Participation in Human Papillomavirus (HPV) vaccination and Papanicolaou Screening (Pap smears) is low among ethnic minorities in the Netherlands and hardly any information is available about the cervical cancer prevention methods of Somali women living in the diaspora. This qualitative study, based on the Health Belief Model (HBM) and an intersectionality-based framework, explores the perceptions of Somali women living in the Netherlands regarding measures to prevent cervical cancer. Semi-structured interviews have been conducted with young Somali women aged 17-21 years (n = 14) and Somali mothers aged 30-46 years (n = 6). Two natural group discussions have been conducted with 12 and 14 Somali mothers aged 23-66 years. The collected data has been analyzed thematically for content. In this study, we have identified perceived barriers to the use of preventive measures across three major themes: (1) Somali women and preventive healthcare; (2) Language, knowledge, and negotiating decisions; and (3) Sexual standards, culture, and religion. Many issues have been identified across these themes, e.g., distrust of the Dutch health care system or being embarrassed to get Pap smears due to Female Genital Mutilation (FGM) and having a Dutch, male practitioner; or a perceived low susceptibility to HPV and cancer because of the religious norms that prohibit sex before marriage. Current measures in the Netherlands to prevent women from developing cervical cancer hardly reach Somali women because these women perceive these kinds of preventative measures as not personally relevant. Dutch education strategies about cervical cancer deviate from ways of exchanging information within the Somali community. Teachers can provide culturally sensitive information to young Somali women in schools. For Somali mothers, oral education (e.g., poetry or theater) about the Dutch health care system and men's roles in HPV transmission may be useful. An intersectional approach, grounded in the HBM, is recommended to promote equal access to preventive health care for Somali women.


The changing character of stepfamilies: implications of cohabitation and nonmarital childbearing.
Divorce, nonmarital childbearing, and cohabitation are reshaping family experience in the United States. Because of these changes, our traditional definitions of families decreasingly capture of the social units of interest. We have noted how a significant proportion of officially defined single-parent families actually are two-parent unmarried families. The present paper expands on this perspective with respect to stepfamilies. We must broaden our definition of stepfamilies to include cohabitations involving a child of only one partner, and must recognize the large role of nonmarital childbearing in the creation of stepfamilies. We find that cohabitation and nonmarital childbearing have been important aspects of stepfamily experience for at least two decades, and that this is increasingly so. To define stepfamilies only in terms of marriage clearly underestimates both the level and the trend in stepfamily experience: when cohabitation is taken into account, about two-fifths of all women and 30% of all children are likely to spend some time in a stepfamily.


Cardiac conduction is required to preserve cardiac chamber morphology.
Electrical cardiac forces have been previously hypothesized to play a significant role in cardiac morphogenesis and remodeling. In response to electrical forces, cultured cardiomyocytes rearrange their cytoskeletal structure and modify their gene expression profile. To translate such in vitro data to the intact heart, we used a collection of zebrafish cardiac mutants and transgenics to investigate whether cardiac conduction could influence in vivo cardiac morphogenesis independent of contractile forces. We show that the cardiac mutant dco(s226) develops heart failure and interrupted cardiac morphogenesis following uncoordinated ventricular contraction. Using in vivo optical mapping/calcium imaging, we determined that the dco cardiac phenotype was primarily due to aberrant ventricular conduction. Because cardiac contraction and intracardiac hemodynamic forces can also influence cardiac development, we further analyzed the dco phenotype in noncontractile hearts and observed that disorganized ventricular conduction could affect cardiomyocyte morphology and subsequent heart morphogenesis in the absence of contraction or flow. By positional cloning, we found that dco encodes Gja3/Cx46, a gap junction protein not previously implicated in heart formation or function. Detailed analysis of the mouse Cx46 mutant revealed the presence of cardiac conduction defects frequently associated with human heart failure. Overall, these in vivo studies indicate that cardiac electrical forces are required to preserve cardiac chamber morphology and may act as a key epigenetic factor in cardiac remodeling.


Vitamin B12 deficiency in patients with chronic-tinnitus and noise-induced hearing loss.
This study examines the incidence of vitamin B12 deficiency in three groups of noise-exposed subjects: patients with chronic tinnitus and noise-induced hearing loss (NIHL), patients with NIHL only, and subjects demonstrating normal hearing. A group of 113 army personnel exposed to military noise was studied. The mean age was 39 years. Chronic tinnitus and NIHL existed in 57 subjects. NIHL alone was observed in 29 subjects, and 27 subjects had normal audiograms. All subjects were queried about noise exposure and dietary habits. Vitamin B12 serum levels were measured. Patients with tinnitus and NIHL exhibited vitamin B12 deficiency in 47% of cases (blood levels < or = 250 pg/mL). This was significantly more (P < .023) compared with NIHL and normal subjects who exhibited vitamin B12 deficiency in 27% and 19%, respectively. These observations suggest a relationship between vitamin B12 deficiency and dysfunction of the auditory pathway. Some improvement in tinnitus and associated complaints were observed in 12 patients following vitamin B12 replacement therapy. The authors recommend that routine vitamin B12 serum levels be determined when evaluating patients for chronic tinnitus.


Preferential desulfurization of dibenzyl sulfide by an isolated Gordonia sp. IITR100.
Several organosulfur compounds are present in the crude oil, and are required to be removed before its processing into transport fuel. For this reason, biodesulfurization of thiophenic compounds has been studied extensively. However, studies on the sulfide compounds are scarce. In this paper, we describe desulfurization of a model sulfidic compound, dibenzyl sulfide (DBS) by an isolated Gordonia sp. IITR100. The reaction was accompanied with the formation of metabolites dibenzyl sulfoxide, dibenzyl sulfone and benzoic acid. Studies with recombinant E. coli revealed that enzyme DszC of this isolate metabolizes DBS into dibenzyl sulfoxide and dibenzyl sulfone, but the reaction downstream to it is mediated by some enzyme other than its DszA. In reactions where DBS and dibenzothiophene (DBT) were present together, both IITR100 and recombinant E. coli exhibited preference for the desulfurization of DBS over DBT. The newly identified capability of IITR100 for desulfurization of both thiophenic and sulfidic compounds suggests its potential use in improved desulfurization of petroleum fractions.


The Impact of Weight Gain During HIV Treatment on Risk of Pre-diabetes, Diabetes Mellitus, Cardiovascular Disease, and Mortality.
Since the introduction of combined antiretroviral therapy (cART) and more effective treatments for AIDS, there has been a dramatic shift from the weight loss and wasting that characterised HIV/AIDS (and still does in countries where cART is not readily available or is initiated late) to healthy weight, or even overweight and obesity at rates mirroring those seen in the general population. These trends are attributable to several factors, including the "return to health" weight gain with reversal of the catabolic effects of HIV-infection following cART-initiation, strategies for earlier cART-initiation in the course of HIV-infection which have prevented many people living with HIV-infection from developing wasting, in addition to exposure to the modern obesogenic environment. Older cART regimens were associated with increased risk of body fat partitioning disorders (lipodystrophy) and cardiometabolic complications including atherothrombotic cardiovascular disease (CVD) and diabetes mellitus. Whilst cART now avoids those medications implicated in causing lipodystrophy, long-term cardiometabolic data on more modern cART regimens are lacking. Longitudinal studies show increased rates of incident CVD and diabetes mellitus with weight gain in treated HIV-infection. Abdominal fat gain, weight gain, and rising body mass index (BMI) in the short-term during HIV treatment was found to increase incident diabetes risk. Rising BMI was associated with increased risk of incident CVD, however the relationship varied depending on pre-cART BMI category. In contrast, a protective association with mortality is evident, predominantly in the underweight and in resource-poor settings, where weight gain reflects access to cART and virological suppression. The question of how to best evaluate, manage (and perhaps constrain) weight gain during HIV treatment is of clinical relevance, especially in the current climate of increasingly widespread cART use, rising overweight, and obesity prevalence and growing metabolic and cardiovascular disease burden in people living with HIV-infection. Large prospective studies to further characterise the relationship between weight gain during HIV treatment and risk of diabetes, CVD and mortality are required.


Combined analysis of three studies of patients with Burkitt's lymphoma.
Data from the three preceding studies on the possible relationship of HL-A type with Burkitt's lymphoma were combined to see whether any trends in the data, not significant in the individal studies, would show significance when the three groups were combined. A statistical method for combining data from different studies is developed. Although in this analysis even the combined data failed to show any significant association of HL-A type with disease, it is suggested that the statistical methods used will be helpful in analysing other sets of data where it seems desirable to combine information from several studies.


Optical diode using nonlinear polystyrene ring resonators in two-dimensional photonic crystal structure.
An optical diode that uses nonlinear ring resonators in 2D photonic crystal is numerically simulated by using the finite-difference time-domain (FDTD) method. Nonlinear polystyrene is used as the Kerr medium forming ring resonators. The operating wavelength of the optical diode is considered to be the coupling wavelength at which light couples efficiently from waveguide to ring resonator, which is also equal to the average of the resonant wavelengths of the two resonators considered in the proposed structure. For both forward and backward propagation, the characteristics of the proposed optical diode are similar to those of an electronic diode. FDTD simulation is done using the MEEP package, which exhibits the desired results.


Changing bodies changes minds: owning another body affects social cognition.
Research on stereotypes demonstrates how existing prejudice affects the way we process outgroups. Recent studies have considered whether it is possible to change our implicit social bias by experimentally changing the relationship between the self and outgroups. In a number of experimental studies, participants have been exposed to bodily illusions that induced ownership over a body different to their own with respect to gender, age, or race. Ownership of an outgroup body has been found to be associated with a significant reduction in implicit biases against that outgroup. We propose that these changes occur via a process of self association that first takes place in the physical, bodily domain as an increase in perceived physical similarity between self and outgroup member. This self association then extends to the conceptual domain, leading to a generalization of positive self-like associations to the outgroup.


p16/Ki-67 Dual Stain Cytology for Detection of Cervical Precancer in HPV-Positive Women.
Human papillomavirus (HPV)-based cervical cancer screening requires triage markers to decide who should be referred to colposcopy. p16/Ki-67 dual stain cytology has been proposed as a biomarker for cervical precancers. We evaluated the dual stain in a large population of HPV-positive women. One thousand five hundred and nine HPV-positive women screened with HPV/cytology cotesting at Kaiser Permanente California were enrolled into a prospective observational study in 2012. Dual stain cytology was performed on residual Surepath material, and slides were evaluated for dual stain-positive cells. Disease endpoints were ascertained from the clinical database at KPNC. We evaluated the clinical performance of the assay among all HPV-positive women and among HPV-positive, cytology-negative women. We used internal benchmarks for clinical management to evaluate the clinical relevance of the dual stain assay. We evaluated sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the dual stain compared with Pap cytology. All statistical tests were two-sided. The dual stain had lower positivity (45.9%) compared with cytology at an ASC-US threshold (53.4%). For detection of CIN2+, the dual stain had similar sensitivity (83.4% vs 76.6%, P = .1), and statistically higher specificity (58.9% vs 49.6%, P < .001), PPV (21.0% vs 16.6%, P < .001), and NPV (96.4% vs 94.2%, P = .01) compared with cytology. Similar patterns were observed for CIN3+. Women with a positive test had high enough risk for referral to colposcopy, while the risk for women with negative tests was below a one-year return threshold based on current US management guidelines. Dual stain cytology showed good risk stratification for all HPV-positive women and for HPV-positive women with normal cytology. Additional follow-up is needed to determine how long dual stain negative women remain at low risk of precancer.


Allometry in desert ant locomotion (Cataglyphis albicans and Cataglyphis bicolor) - does body size matter?
Desert ants show a large range of adaptations to their habitats. They can reach extremely high running speeds, for example, to shorten heat stress during foraging trips. It has recently been examined how fast walking speeds are achieved in different desert ant species. It is intriguing in this context that some species exhibit distinct intraspecific size differences. We therefore performed a complete locomotion analysis over the entire size spectrum of the species Cataglyphis bicolor, and we compared this intraspecific dataset with that of the allometrically similar species Cataglyphis albicans. Emphasis was on the allometry of locomotion: we considered the body size of each animal and analysed the data in terms of relative walking speed. Body size was observed to affect walking parameters, gait patterns and phase relationships in terms of absolute walking speed. Unexpectedly, on a relative scale, all ants tended to show the same overall locomotion strategy at low walking speeds, and significant differences occurred only between C. albicans and C. bicolor at high walking speeds. Our analysis revealed that C. bicolor ants use the same overall strategy across all body sizes, with small ants reaching the highest walking speeds (up to 80 body lengths s-1) by increasing their stride length and incorporating aerial phases. By comparison, C. albicans reached high walking speeds mainly by a high synchrony of leg movement, lower swing phase duration and higher stride frequency ranging up to 40 Hz.


Genetic variation within a dominant shrub structures green and brown community assemblages.
Two rising challenges in ecology are understanding the linkages between above- and belowground components of terrestrial ecosystems and connecting genes to their ecological consequences. Here, we blend these emerging perspectives using a long-term common-garden experiment in a coastal dune ecosystem, whose dominant shrub species, Baccharis pilularis, exists as erect or prostrate architectural morphotypes. We explored variation in green (foliage-based) and brown (detritus-based) community assemblages, local ecosystem processes, and understory microclimate between the two morphs. Prostrate morphs supported more individuals, species, and different compositions of foliage arthropods, litter microarthropods, and soil bacteria than erect morphs. The magnitude of community compositional differences was maintained from crown to litter to soil. Despite showing strikingly similar responses, green and brown assemblages were associated with different underlying mechanisms. Differences in estimated shrub biomass best explained variation in the green assemblage, while understory abiotic conditions accounted for variation in the brown assemblage. Prostrate morphs produced more biomass and litter, which corresponded with their strong lateral growth in a windy environment. Compared to erect morphs, the denser canopy and thicker litter layer of prostrate morphs helped create more humid understory conditions. As a result, decomposition rates were higher under prostrate shrubs, despite prostrate litter being of poorer quality. Together, our results support the hypothesis that intraspecific genetic variation in primary producers is a key mediator of above- and belowground linkages, and that integrating the two perspectives can lead to new insights into how terrestrial communities are linked with ecosystem pools and processes.