Activation of cGMP-dependent protein kinase stimulates cardiac ATP-sensitive potassium channels via a ROS/calmodulin/CaMKII signaling cascade.
Cyclic GMP (cGMP)-dependent protein kinase (PKG) is recognized as an important signaling component in diverse cell types. PKG may influence the function of cardiac ATP-sensitive potassium (K(ATP)) channels, an ion channel critical for stress adaptation in the heart; however, the underlying mechanism remains largely unknown. The present study was designed to address this issue. Single-channel recordings of cardiac K(ATP) channels were performed in both cell-attached and inside-out patch configurations using transfected human embryonic kidney (HEK)293 cells and rabbit ventricular cardiomyocytes. We found that Kir6.2/SUR2A (the cardiac-type K(ATP)) channels were activated by cGMP-selective phosphodiesterase inhibitor zaprinast in a concentration-dependent manner in cell-attached patches obtained from HEK293 cells, an effect mimicked by the membrane-permeable cGMP analog 8-bromo-cGMP whereas abolished by selective PKG inhibitors. Intriguingly, direct application of PKG moderately reduced rather than augmented Kir6.2/SUR2A single-channel currents in excised, inside-out patches. Moreover, PKG stimulation of Kir6.2/SUR2A channels in intact cells was abrogated by ROS/H(2)O(2) scavenging, antagonism of calmodulin, and blockade of calcium/calmodulin-dependent protein kinase II (CaMKII), respectively. Exogenous H(2)O(2) also concentration-dependently stimulated Kir6.2/SUR2A channels in intact cells, and its effect was prevented by inhibition of calmodulin or CaMKII. PKG stimulation of K(ATP) channels was confirmed in intact ventricular cardiomyocytes, which was ROS- and CaMKII-dependent. Kinetically, PKG appeared to stimulate these channels by destabilizing the longest closed state while stabilizing the long open state and facilitating opening transitions. The present study provides novel evidence that PKG exerts dual regulation of cardiac K(ATP) channels, including marked stimulation resulting from intracellular signaling mediated by ROS (H(2)O(2) in particular), calmodulin and CaMKII, alongside of moderate channel suppression likely mediated by direct PKG phosphorylation of the channel or some closely associated proteins. The novel cGMP/PKG/ROS/calmodulin/CaMKII signaling pathway may regulate cardiomyocyte excitability by opening K(ATP) channels and contribute to cardiac protection against ischemia-reperfusion injury.


Black and Minority Ethnic women's decision-making for risk reduction strategies after BRCA testing: Use of context and knowledge.
Within the field of breast cancer care, women concerned about their family history are offered genetic testing and subsequent treatment options based on several factors which include but are not limited to personal and family cancer disease histories and clinical guidelines. Discussions around decision-making in genetics in Black and Minority Ethnic (BME) groups are rarely documented in literature, and information regarding interactions with genetics services is usually discussed and linked to lack of scientific knowledge. As such, counselling sessions based only on scientific and medical information miss out the many reasons participants consider in making health decisions, information which can be used to encourage BME women to engage in cancer genetics services. 10 BME women with a mixed personal and family history of breast and ovarian cancer backgrounds, were interviewed in a study exploring issues of knowledge about familial breast cancer syndromes, to understand how they created and used familial knowledge for health decisions, with a particular focus on attitudes towards risk reducing strategies. Study results show that our participants are not unique in the ways they make decisions towards the use of cancer genetics and risk reduction strategies and as such, there are no specific ethnically defined pathways for decision-making. Our participants demonstrated mixed biomedical, social and individual cultural reasons for their decision-making towards risk reduction surgeries and treatment options which are similar to women from different ethnicities and are individual rather than group-specific. Narratives about suspicion of scientific utility of genetic knowledge, the perceived predictive value of mutations for future cancers or the origin of mutations and family disease patterns feature heavily in how participants evaluated genetic information and treatment decisions. The diversity of results shows that our participants are interested in engaging with genetic information but use multiple sources for evaluating the extent of involvement in genetic services and the place of genetic information and treatment options for themselves and their families. Genetic information is considered within various bio-social scenarios before decision-making for risk reduction is undertaken. BME women are shown to undertake evaluative processes which clinicians are encouraged to explore for better patient support. Continuing to focus on links between superficial and un-representative meanings of ethnicity, ethnic identity and attitudes and behaviours by only searching for differences between ethnic groups, are unhelpful in further understanding how women from those diverse backgrounds make decisions towards risk reduction interventions. Future research must find ways of investigating and understanding populations in ways that are not focussed solely on ethnic differences but on how meaning is created out of social circumstances and experiences.


Validated Risk Score for Predicting 6-Month Mortality in Infective Endocarditis.
Host factors and complications have been associated with higher mortality in infective endocarditis (IE). We sought to develop and validate a model of clinical characteristics to predict 6-month mortality in IE. Using a large multinational prospective registry of definite IE (International Collaboration on Endocarditis [ICE]-Prospective Cohort Study [PCS], 2000-2006, n=4049), a model to predict 6-month survival was developed by Cox proportional hazards modeling with inverse probability weighting for surgery treatment and was internally validated by the bootstrapping method. This model was externally validated in an independent prospective registry (ICE-PLUS, 2008-2012, n=1197). The 6-month mortality was 971 of 4049 (24.0%) in the ICE-PCS cohort and 342 of 1197 (28.6%) in the ICE-PLUS cohort. Surgery during the index hospitalization was performed in 48.1% and 54.0% of the cohorts, respectively. In the derivation model, variables related to host factors (age, dialysis), IE characteristics (prosthetic or nosocomial IE, causative organism, left-sided valve vegetation), and IE complications (severe heart failure, stroke, paravalvular complication, and persistent bacteremia) were independently associated with 6-month mortality, and surgery was associated with a lower risk of mortality (Harrell's C statistic 0.715). In the validation model, these variables had similar hazard ratios (Harrell's C statistic 0.682), with a similar, independent benefit of surgery (hazard ratio 0.74, 95% CI 0.62-0.89). A simplified risk model was developed by weight adjustment of these variables. Six-month mortality after IE is ≈25% and is predicted by host factors, IE characteristics, and IE complications. Surgery during the index hospitalization is associated with lower mortality but is performed less frequently in the highest risk patients. A simplified risk model may be used to identify specific risk subgroups in IE.


The reach of chronic-disease self-management education programs to rural populations.
This study assessed the sociodemographic characteristics of rural residents who participated in chronic-disease self-management education (CDSME) program workshops and the extent to which CDSME programs were utilized by those with limited access to health care services. We analyzed data from the first 100,000 adults who attended CDSME program workshops during a national dissemination spanning 45 states, the District of Columbia, and Puerto Rico. Approximately 24% of participants lived in rural areas. Overall, 42% of all participants were minorities; urban areas reached more minority participants (48%) than rural areas (25%). The average age of participants was high in rural (age, μ = 66.1) and urban (age, μ = 67.3) areas. In addition, the average number of chronic conditions was higher (p < 0.01) in rural (μ = 2.6 conditions) versus urban (μ = 2.4 conditions) areas. Successful completion of CDSME programs (i.e., attending four or more of the six workshop sessions) was higher (p < 0.01) in rural versus urban areas (78% versus 77%). Factors associated with higher likelihood of successful completion of CDSME programs included being Black (OR = 1.25) versus White and living in rural (versus urban) areas (OR = 1.09). Factors associated with lower likelihood of successful completion included being male (OR = 0.92) and residing in a primary care Health Professional Shortage Area or HPSA (versus a non-HPSA) (OR = 0.93). Findings highlight the capability of CDSME programs to reach rural residents, yet dissemination efforts can be further enhanced to ensure minorities and individuals in a HPSA utilize this program. Tailored strategies are needed to increase participant recruitment and retention in rural areas to overcome traditional barriers to health service access.


Fifteen years of heroin-assisted treatment in a Swiss prison-a retrospective cohort study.
In the context of the current US opioid crisis and the compelling fact that a quarter to a third of all those addicted to heroin pass through its prisons and jails each year, the care of incarcerated opioid-using individuals (OUI) needs to be improved. Little has been published on the effectiveness or outcomes of heroin-assisted treatment (HAT), a treatment option for severely dependent OUI delivered in a prison setting. The aim of this study was therefore to evaluate such treatment since its implementation. The primary objective was to investigate whether heroin-assisted treatment was associated with severe detrimental health outcomes. The secondary objective was to compare the heroin-assisted treatment group with the general prison population in terms of occupational functioning. Retrospective cohort study SETTING: An open prison with 120 places SUBJECTS: Data on 1885 male prisoners with a total of 2239 imprisonment periods between 2000 and 2015 was available. Ninety-seven inmates in heroin-assisted treatment were compared with 1788 inmates from the general prison population (reference group). Mortality, medical complications (including overdoses), and work performance (days worked, sick days, and monthly wages earned). Inmates receiving HAT were on average 1 year younger (33.8 vs. 34.9 years), had longer prison stays (7.3 vs. 3.0 months), were more often of Swiss nationality (68.0% vs. 28.9%), and had committed more drug- and property-related offenses (49.5% vs. 23.2% and 63.9% vs. 38.3%, respectively) compared to the reference group. No serious heroin-related medical complication occurred during the 15-year window of observation among inmates with heroin-assisted treatment. Their work performance was comparable to that of the reference group. This study shows that heroin-assisted treatment can be a valuable treatment option for severely dependent OUI during imprisonment, can be delivered safely by prison health staff over extended periods of time, and allows OUI in treatment to achieve work performance rates comparable to that of the general prison population.


Development of an intervention to reduce antibiotic use for childhood coughs in UK primary care using critical synthesis of multi-method research.
Overuse of antibiotics contributes to the global threat of antimicrobial resistance. Antibiotic stewardship interventions address this threat by reducing the use of antibiotics in occasions or doses unlikely to be effective. We aimed to develop an evidence-based, theory-informed, intervention to reduce antibiotic prescriptions in primary care for childhood respiratory tract infections (RTI). This paper describes our methods for doing so. Green and Krueter's Precede/Proceed logic model was used as a framework to integrate findings from a programme of research including 5 systematic reviews, 3 qualitative studies, and 1 cohort study. The model was populated using a strength of evidence approach, and developed with input from stakeholders including clinicians and parents. The synthesis produced a series of evidence-based statements summarizing the quantitative and qualitative evidence for intervention elements most likely to result in changes in clinician behaviour. Current evidence suggests that interventions which reduce clinical uncertainty, reduce clinician/parent miscommunication, elicit parent concerns, make clear delayed or no-antibiotic recommendations, and provide clinicians with alternate treatment actions have the best chance of success. We designed a web-based within-consultation intervention to reduce clinician uncertainty and pressure to prescribe, designed to be used when children with RTI present to a prescribing clinician in primary care. We provide a worked example of methods for the development of future complex interventions in primary care, where multiple factors act on multiple actors within a complex system. Our synthesis provided intervention guidance, recommendations for practice, and highlighted evidence gaps, but questions remain about how best to implement these recommendations. The funding structure which enabled a single team of researchers to work on a multi-method programme of related studies (NIHR Programme Grant scheme) was key in our success. The feasibility study accompanying this intervention was prospectively registered with the ISRCTN registry ( ISRCTN23547970 ), on 27 June 2014.


Community participation in New Mexico's behavioral health care reform.
In 2005, New Mexico implemented a unique reform in managed behavioral health services that seeks to ensure delivery of consumer-driven, recovery-oriented care to low-income individuals. Distinguishing features of the reform are the Local Collaboratives (LCs), regionally based community organizations designed by state government to represent behavioral health concerns of New Mexico's diverse cultural populations. We examine community response to the LCs, focusing on two broad sets of themes derived from 18 months of ethnographic fieldwork. The first set--structure and function--encompasses several issues: predominance of provider versus consumer voice; insufficient resources to support internal operations; imposition of state administrative demands; and perceived lack of state response to LC efforts. The second set--participation and collaboration--reveals how problems of information flow and other logistical factors impact involvement in LCs and how the construction of "community" introduced under this initiative exacerbates tensions across localities with varied histories and populations.


Adaptively modulated optical OFDM modems utilizing RSOAs as intensity modulators in IMDD SMF transmission systems.
Detailed investigations of the transmission performance of adaptively modulated optical orthogonal frequency division multiplexed (AMOOFDM) signals converted using reflective semiconductor optical amplifiers (RSOAs) are undertaken over intensity-modulation and direct-detection (IMDD) single-mode fiber (SMF) transmission systems for WDM-PONs. The theoretical RSOA model adopted for modulating the AMOOFDM signals is experimentally verified rigorously in the aforementioned transmission systems incorporating recently developed real-time end-to-end OOFDM transceivers. Extensive performance comparisons are also made between RSOA and SOA intensity modulators. Optimum RSOA operating conditions are identified, which are independent of RSOA rear-facet reflectivity and very similar to those corresponding to SOAs. Under the identified optimum operating conditions, the RSOA and SOA intensity modulators support the identical AMOOFDM transmission performance of 30Gb/s over 60km SMFs. Under low-cost optical component-enabled practical operating conditions, RSOA intensity modulators with rear-facet reflectivity values of >0.3 outperform considerably SOA intensity modulators in transmission performance, which decreases significantly with reducing RSOA rear-facet reflectivity and optical input power. In addition, results also show that use can be made of the RSOA/SOA intensity modulation-induced negative frequency chirp to improve the AMOOFDM transmission performance in IMDD SMF systems.


Syntheses and characterization of copper(II) carboxylate dimers formed from enantiopure ligands containing a strong π···π stacking synthon: enantioselective single-crystal to single-crystal gas/solid-mediated transformations.
Tri- and tetrafunctional enantiopure ligands have been prepared from 1,8-naphthalic anhydride and the amino acids L-alanine, D-phenylglycine, and L-asparagine to produce (S)-2-(1,8-naphthalimido)propanoic acid (HL(ala)), (R)-2-(1,8-naphthalimido)-2-phenylacetic acid (HL(phg)), and (S)-4-amino-2-(1,8 naphthalimido)-4-oxobutanoic acid (HL(asn)), respectively. Reactions of L(ala)(-) with copper(II) acetate under a variety of solvent conditions has led to the formation and characterization by X-ray crystallography of three similar copper(II) paddlewheel complexes with different axial ligands, [Cu(2)(L(ala))(4)(THF)(2)] (1), [Cu(2)(L(ala))(4)(HL(ala))] (2), and [Cu(2)(L(ala))(4)(py)(THF)] (3). A similar reaction using THF and L(phg)(-) leads to the formation of [Cu(2)(L(phg))(4)(THF)(2)] (4). With the exception of a disordered component in the structure of 4, the naphthalimide groups in all of these compounds are arranged on the same side of the square, central paddlewheel unit, forming what is known as the chiral crown configuration. A variety of π···π stacking interactions of the 1,8-naphthalimide groups organize all of these complexes into supramolecular structures. The addition of the amide group functionality in the L(asn)(-) ligand leads to the formation of tetrameric [Cu(4)(L(asn))(8)(py)(MeOH)] (5), where reciprocal axial coordination of one of the amide carbonyl oxygen atoms between two dimers leads to the tetramer. Extensive supramolecular interactions in 5, mainly the π···π stacking interactions of the 1,8-naphthalimide supramolecular synthon, support an open three-dimensional structure containing large pores filled with solvent. When crystals of [Cu(4)(L(asn))(8)(py)(MeOH)] are exposed to (S)-ethyl lactate vapor, the coordinated methanol molecule is replaced by (S)-ethyl lactate, bonding to the copper ion through the carbonyl oxygen, yielding [Cu(4)(L(asn))(8)(py)((S)-ethyl lactate)] (6) without a loss of crystallinity. With the exception of the replacement of the one axial ligand, the molecular structures of 5 and 6 are very similar. In a similar experiment of 5 with vapors of (R)-ethyl lactate, again a change occurs without a loss of crystallinity, but in this case the (R)-ethyl lactate displaces only slightly more than half of the axial methanol molecules forming [Cu(4)(L(asn))(8)(py){((R)-ethyl lactate)(0.58)(MeOH)(0.42)}] (7). Importantly, in 7, the (R)-ethyl lactate coordinates through the hydroxyl group. When crystals of [Cu(4)(L(asn))(8)(py)(MeOH)] are exposed to vapors of racemic ethyl lactate, the coordinated methanol molecule is displaced without a loss of crystallinity exclusively by (S)-ethyl lactate, yielding a new form of the tetramer [Cu(4)(L(asn))(8)(py)((S)-ethyl lactate)], in which the ethyl lactate in the pocket bonds to the copper(II) ion through the carbonyl oxygen as with 6. Exposure of [Cu(4)(L(asn))(8)(py){((R)-ethyl lactate)(0.58)(MeOH)(0.42)}] to racemic ethyl lactate yields a third form of [Cu(4)(L(asn))(8)(py)((S)-ethyl lactate)], where the three forms of [Cu(4)(L(asn))(8)(py)((S)-ethyl lactate)] have differences in the number of ordered (S)-ethyl lactate molecules located in the interstitial sites. These results demonstrate enantioselective bonding to a metal center in the chiral pocket of both 5 and 7 during single-crystal to single-crystal gas/solid-mediated exchange reactions.


Aberration correction in an adaptive free-space optical interconnect with an error diffusion algorithm.
Aberration correction within a free-space optical interconnect based on a spatial light modulator for beam steering and holographic wavefront correction is presented. The wavefront sensing technique is based on an extension of a modal wavefront sensor described by Neil et al. [J. Opt. Soc. Am. A 17, 1098 (2000)], which uses a diffractive element. In this analysis such a wavefront sensor is adapted with an error diffusion algorithm that yields a low reconstruction error and fast reconfigurability. Improvement of the beam propagation quality (Strehl ratio) for different channels across the input plane is achieved. However, due to the space invariancy of the system, a trade-off among the beam propagation quality for channels is obtained. Experimental results are presented and discussed.


Bmi1 confers resistance to oxidative stress on hematopoietic stem cells.
The polycomb-group (PcG) proteins function as general regulators of stem cells. We previously reported that retrovirus-mediated overexpression of Bmi1, a gene encoding a core component of polycomb repressive complex (PRC) 1, maintained self-renewing hematopoietic stem cells (HSCs) during long-term culture. However, the effects of overexpression of Bmi1 on HSCs in vivo remained to be precisely addressed. In this study, we generated a mouse line where Bmi1 can be conditionally overexpressed under the control of the endogenous Rosa26 promoter in a hematopoietic cell-specific fashion (Tie2-Cre;R26Stop(FL)Bmi1). Although overexpression of Bmi1 did not significantly affect steady state hematopoiesis, it promoted expansion of functional HSCs during ex vivo culture and efficiently protected HSCs against loss of self-renewal capacity during serial transplantation. Overexpression of Bmi1 had no effect on DNA damage response triggered by ionizing radiation. In contrast, Tie2-Cre;R26Stop(FL)Bmi1 HSCs under oxidative stress maintained a multipotent state and generally tolerated oxidative stress better than the control. Unexpectedly, overexpression of Bmi1 had no impact on the level of intracellular reactive oxygen species (ROS). Our findings demonstrate that overexpression of Bmi1 confers resistance to stresses, particularly oxidative stress, onto HSCs. This thereby enhances their regenerative capacity and suggests that Bmi1 is located downstream of ROS signaling and negatively regulated by it.


Harnessing the multifunctionality in nature: a bioactive agent release system with self-antimicrobial and immunomodulatory properties.
Major problems with biomedical devices in particular implants located in nonsterile environments concern: (i) excessive immune response to the implant, (ii) development of bacterial biofilms, and (iii) yeast and fungi infections. An original multifunctional coating that addresses all these issues concomitantly is developed. A new exponentially growing polyelectrolyte multilayer film based on polyarginine (PAR) and hyaluronic acid (HA) is designed. The films have a strong inhibitory effect on the production of inflammatory cytokines released by human primary macrophage subpopulations. This could reduce potential chronic inflammatory reaction following implantation. Next, it is shown that PAR, due to its positive charges, has an antimicrobial activity in film format against Staphylococcus aureus for 24 h. In order to have a long-term antimicrobial activity, a precursor nanoscale silver coating is deposited on the surface before adding the PAR/HA films. Moreover, the PAR/HA films can be easily further functionalized by embedding antimicrobial peptides, like catestatin (CAT), a natural host defense peptide. This PAR/HA+CAT film proves to be effective as an antimicrobial coating against yeast and fungi and its cytocompatibility is also assessed. Finally, this all-in-one system constitutes an original strategy to limit inflammation and prevents bacteria, yeast, and fungi infections.


Under-reporting of maritime accidents.
The majority of current maritime regulations has been developed following a reactive approach, often as ad-hoc response to serious accidents, and are characterised as being prescriptive leaving limited space for adapting equivalent solutions rather those described in the regulations. On the premise of providing a more proactive approach for the proposal or the evaluation of regulations, the Formal Safety Assessment (FSA) has been introduced. In the context of FSA, the analysis of accident data is considered to be very important for providing potential input on developing more balanced, proactive and cost-effective regulations. However, it has been argued that the validity of historical data may be undermined by uncertainties. This paper is aimed at showing evidence on serious under-reporting in accident databases, which can be considered as the main contributor to questioning the direct and uncritical use of historical data. By analysing the 10-year tanker accident data from the Lloyd's Register FairPlay (LRFP) and the Norwegian Maritime Directorate (NMD) for vessels registered in Norway, it is found that the reporting performance has an upper bound of 41% for NMD and 30% for LRFP. Furthermore, based on comparison between LRFP data and self-assessment by Flag States, it is seen that accidents reported by the Flag States are also incomplete.


Modulation of aberrant splicing in human RNA diseases by chemical compounds.
Pre-mRNA splicing is an essential step for gene expression in higher eukaryotes. Alternative splicing contributes to diversity of the expressed proteins from the limited number of genes. Disruption of splicing regulation often results in hereditary and sporadic diseases called as 'RNA diseases'. Modulation of splicing by small chemical compounds and nucleic acids has been tried to target aberrant splicing in those diseases. Several RNA diseases and splicing-target therapeutic approaches will be briefly introduced in this review. Accumulating knowledge about molecular mechanism of aberrant splicing and their correction by chemical compounds is important not only for RNA biologists, but also for clinicians who desire therapies for those diseases.


Nurses' emotional experience of caring for children with burns.
The aim of this phenomenological study was to explore the emotions experienced by children's nurses when caring for children with burns, in addition to ascertaining how the nurses dealt with these emotions. The nature of nursing practice is such that it inevitably generates some form of emotional response in nurses. The literature reveals that the manner nurses deal with their emotional experiences can impact on their nursing care. The study used Husserlian phenomenology to explore the emotional experiences of eight purposively selected children's nurses who have worked on the burns unit of an Irish paediatric hospital. Data were collected using in-depth, unstructured interviews and analysed using Colaizzi's seven stage framework. The phenomenon of participants' emotional experiences is captured in four themes: (1) caring for children with burns, (2) supporting parents, (3) sustaining nurses' emotional well-being, and (4) learning to be a burns nurse. Nursing children with burns generated a myriad of emotions for participants. Burns dressing-changes, managing burn-related pain, supporting parents and the impact of busy workloads on the emotional care of children and their parents emerged as the most emotionally challenging aspects of participants' role. Participants recognised the need to manage their emotional responses and spoke of the benefits of a supportive nursing team. The findings offer insights into both the rewarding and challenging aspects of nursing children with burns. Nurses in this environment must be supported to recognise and manage their emotional responses to their work. Helping nurses to manage the emotional consequences of their work will help to sustain their emotional well-being, enhance the care received by children and also enable nurses to support parents in their role as partners in care.


Integrating acoustic telemetry into mark-recapture models to improve the precision of apparent survival and abundance estimates.
Capture-mark-recapture models are useful tools for estimating demographic parameters but often result in low precision when recapture rates are low. Low recapture rates are typical in many study systems including fishing-based studies. Incorporating auxiliary data into the models can improve precision and in some cases enable parameter estimation. Here, we present a novel application of acoustic telemetry for the estimation of apparent survival and abundance within capture-mark-recapture analysis using open population models. Our case study is based on simultaneously collecting longline fishing and acoustic telemetry data for a large mobile apex predator, the broadnose sevengill shark (Notorhynchus cepedianus), at a coastal site in Tasmania, Australia. Cormack-Jolly-Seber models showed that longline data alone had very low recapture rates while acoustic telemetry data for the same time period resulted in at least tenfold higher recapture rates. The apparent survival estimates were similar for the two datasets but the acoustic telemetry data showed much greater precision and enabled apparent survival parameter estimation for one dataset, which was inestimable using fishing data alone. Combined acoustic telemetry and longline data were incorporated into Jolly-Seber models using a Monte Carlo simulation approach. Abundance estimates were comparable to those with longline data only; however, the inclusion of acoustic telemetry data increased precision in the estimates. We conclude that acoustic telemetry is a useful tool for incorporating in capture-mark-recapture studies in the marine environment. Future studies should consider the application of acoustic telemetry within this framework when setting up the study design and sampling program.


Non-inferiority trials: No better? No worse? No change? No pain?
With improvements in care over time it becomes harder to improve clinical outcomes in those conditions where cure rates are high. The focus of research can thus turn to the so-called non-inferiority trial: where the main aim is not to improve clinical outcome, but instead to provide evidence of a lack of difference, whilst other issues, such as cost or toxicity, are improved. The interpretation of such trials is not always straightforward. The burden of proof is reversed compared to a traditional superiority trial, and this means that many of the statistical safeguards, such as significance and intention-to-treat, which act as restraints from an overhasty adoption of a new therapy, may actually work in the opposite fashion. The issues regarding non-inferiority and equivalence trials are considered, and their interpretation discussed.


Factors Influencing Burden in Spouse-Caregivers of Patients with Chronic-Acquired Brain Injury.
In this cross-sectional study, we assess associated factors of burden in spouse-caregivers of patients with acquired brain injury (ABI) in the chronic phase. 35 spouse-caregivers (71% female, mean age ± SD: 55.7 ± 11.1y) of patients with mild/moderate ABI (29% female, mean age ± SD: 57.5 ± 10.7y), admitted to the intensive rehabilitation unit of the Institute S. Anna (Crotone, Italy) between January 2013 and December 2017, were contacted 2 years postinjury and asked to complete a series of questionnaires. The outcome measure was the Caregiver Burden Inventory (CBI) test, while several demographical and clinical data were considered as predictive factors. Two years after injury, a high level of burden was reported in 34.2% of spouse-caregivers. Stepwise multiple linear regression analyses revealed that caring for a patient with more severe disability (as measured by the Barthel Index scale) and the family life cycle (from the initial phase of engagement to marriage with adult children) explain the vast majority of variance for higher caregiver burden. The functional clinical status and the stages through which a family may pass over time were identified as areas in which the spouse-caregiver of ABI patients experienced high levels of burden in the chronic phase.


Fatty acids and inflammation: the cutting edge between food and pharma.
Inflammation underlies many common conditions and diseases. Fatty acids can influence inflammation through a variety of mechanisms, including acting via cell surface and intracellular receptors/sensors that control inflammatory cell signalling and gene expression patterns. Some effects of fatty acids on inflammatory cells appear to be mediated by, or at least are associated with, changes in fatty acid composition of cell membranes. Changes in these compositions can modify membrane fluidity, lipid raft formation, cell signalling leading to altered gene expression, and the pattern of lipid and peptide mediator production. Cells involved in the inflammatory response are typically rich in the n-6 fatty acid arachidonic acid, but the contents of arachidonic acid and of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) can be altered through oral administration of EPA and DHA. Eicosanoids produced from arachidonic acid have roles in inflammation. EPA also gives rise to eicosanoids and these may have differing properties from those of arachidonic acid-derived eicosanoids. EPA and DHA give rise to resolvins which are anti-inflammatory and inflammation resolving. Thus, fatty acid exposure and the fatty acid composition of human inflammatory cells influences their function. As a result of their anti-inflammatory actions marine n-3 fatty acids have therapeutic efficacy in rheumatoid arthritis, although benefits in other inflammatory diseases and conditions have not been unequivocally demonstrated. The anti-inflammatory effects of marine n-3 fatty acids may contribute to their protective actions towards atherosclerosis, plaque rupture and cardiovascular mortality. The therapeutic dose of n-3 fatty acids is not clear.


Histocompatible embryonic stem cells by parthenogenesis.
Genetically matched pluripotent embryonic stem (ES) cells generated via nuclear transfer or parthenogenesis (pES cells) are a potential source of histocompatible cells and tissues for transplantation. After parthenogenetic activation of murine oocytes and interruption of meiosis I or II, we isolated and genotyped pES cells and characterized those that carried the full complement of major histocompatibility complex (MHC) antigens of the oocyte donor. Differentiated tissues from these pES cells engrafted in immunocompetent MHC-matched mouse recipients, demonstrating that selected pES cells can serve as a source of histocompatible tissues for transplantation.


Ionic states of substrates and transition state analogues at the catalytic sites of N-ribosyltransferases.
Purine nucleoside phosphorylase (PNP) and hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) catalyze N-ribosidic bond cleavage in purine nucleosides and nucleotides, with addition of phosphate or pyrophosphate to form phosphorylated alpha-D-ribose products. The transition states have oxacarbenium ion character with a positive charge near 1'-C and ionic stabilization from nearby phosphoryl anions. Immucillin-H (ImmH) and Immucillin-H 5'-PO(4) (ImmHP) resemble the transition state charge when protonated at 4'-N and bind tightly to these enzymes with K(d) values of 20 pM to 1 nM. It has been proposed that Immucillins bind as the 4'-N neutral form and are protonated in the slow-onset step. Solution and solid-state NMR spectra of ImmH, ImmHP, guanosine, and GMP in complexes with two PNPs and a HGPRTase have been used to characterize their ionization states. Results with PNP*ImmH*PO(4) and HGPRTase*ImmHP*MgPP(i) indicate protonation at N-4' for the tightly bound inhibitors. The 1'-(13)C and 1'-(1)H resonances of bound Immucillins showed large downfield shifts as compared to Michaelis complexes, suggesting distortion of 1'-C toward sp(2) geometry. The Immucillins act as transition state mimics by binding with neutral iminoribitol groups followed by 4'-N protonation during slow-onset inhibition to form carbocationic mimics of the transition states. The ability of the Immucillins to mimic both substrate and transition state features contributes to their capture of transition state binding energy. Enzyme-activated phosphoryl nucleophiles bound to PNP and HGPRTase suggest enhanced electrostatic stabilization of the cationic transition states. Distortion of the oxacarbenium ion mimic toward transition state geometry is a common feature of the three distinct enzymatic complexes analyzed here. Substrate complexes, even in catalytically cycling equilibrium mixtures, do not reveal similar distortions.


The VGF-derived Peptide TLQP21 Impairs Purinergic Control of Chemotaxis and Phagocytosis in Mouse Microglia.
Microglial cells are considered as sensors of brain pathology by detecting any sign of brain lesions, infections, or dysfunction and can influence the onset and progression of neurological diseases. They are capable of sensing their neuronal environment via many different signaling molecules, such as neurotransmitters, neurohormones and neuropeptides. The neuropeptide VGF has been associated with many metabolic and neurological disorders. TLQP21 is a VGF-derived peptide and has been shown to signal via C3aR1 and C1qBP receptors. The effect of TLQP21 on microglial functions in health or disease is not known. Studying microglial cells in acute brain slices, we found that TLQP21 impaired metabotropic purinergic signaling. Specifically, it attenuated the ATP-induced activation of a K+ conductance, the UDP-stimulated phagocytic activity, and the ATP-dependent laser lesion-induced process outgrowth. These impairments were reversed by blocking C1qBP, but not C3aR1 receptors. While microglia in brain slices from male mice lack C3aR1 receptors, both receptors are expressed in primary cultured microglia. In addition to the negative impact on purinergic signaling, we found stimulating effects of TLQP21 in cultured microglia, which were mediated by C3aR1 receptors: it directly evoked membrane currents, stimulated basal phagocytic activity, evoked intracellular Ca2+ transient elevations, and served as a chemotactic signal. We conclude that TLQP21 has differential effects on microglia depending on C3aR1 activation or C1qBP-dependent attenuation of purinergic signaling. Thus, TLQP21 can modulate the functional phenotype of microglia, which may have an impact on their function in health and disease.SIGNIFICANCE STATEMENT The neuropeptide VGF and its peptides have been associated with many metabolic and neurological disorders. TLQP21 is a VGF-derived peptide that activates C1qBP receptors, which are expressed by microglia. We show here, for the first time, that TLQP21 impairs P2Y-mediated purinergic signaling and related functions. These include modulation of phagocytic activity and responses to injury. As purinergic signaling is central for microglial actions in the brain, this TLQP21-mediated mechanism might regulate microglial activity in health and disease. We furthermore show that, in addition to C1qBP, functional C3aR1 responses contribute to TLQP21 action on microglia. However, C3aR1 responses were only present in primary cultures but not in situ, suggesting that the expression of these receptors might vary between different microglial activation states.


Advances in T Helper 17 Cell Biology: Pathogenic Role and Potential Therapy in Multiple Sclerosis.
The discovery of the T helper (Th) 17 lineage, involved in the protection against fungal and extracellular bacterial infections, has profoundly revolutionized our current understanding of T cell-mediated responses in autoimmune diseases, including multiple sclerosis (MS). Indeed, recent data demonstrate the pathogenic role of Th17 cells in autoimmune disorders. In particular, studies in MS and in its animal model (EAE, experimental autoimmune encephalomyelitis) have revealed a crucial role of Th17 cells in the pathogenesis of autoimmune demyelinating diseases in both mice and humans. Over the past years, several important aspects concerning Th17 cells have been elucidated, such as the factors which promote or inhibit their differentiation and the effector cytokines which mediate their responses. The identification of the features endowing Th17 cells with high pathogenicity in MS is of particular interest, and discoveries in Th17 cell biology and function could lead to the design of new strategies aimed at modulating the immune response in MS. Here, we will discuss recent advances in this field, with particular focus on the mechanisms conferring pathogenicity in MS and their potential modulation.


Environmental transmission of Toxoplasma gondii: Oocysts in water, soil and food.
Toxoplasma gondii is a zoonotic protozoan parasite that can cause morbidity and mortality in humans, domestic animals, and terrestrial and aquatic wildlife. The environmentally robust oocyst stage of T. gondii is fundamentally critical to the parasite's success, both in terms of its worldwide distribution as well as the extensive range of infected intermediate hosts. Despite the limited definitive host species (domestic and wild felids), infections have been reported on every continent, and in terrestrial as well as aquatic environments. The remarkable resistance of the oocyst wall enables dissemination of T. gondii through watersheds and ecosystems, and long-term persistence in diverse foods such as shellfish and fresh produce. Here, we review the key attributes of oocyst biophysical properties that confer their ability to disseminate and survive in the environment, as well as the epidemiological dynamics of oocyst sources including domestic and wild felids. This manuscript further provides a comprehensive review of the pathways by which T. gondii oocysts can infect animals and people through the environment, including in contaminated foods, water or soil. We conclude by identifying critical control points for reducing risk of exposure to oocysts as well as opportunities for future synergies and new directions for research aimed at reducing the burden of oocyst-borne toxoplasmosis in humans, domestic animals, and wildlife.


Primary care of adult survivors of childhood cancer.
There are approximately 300,000 survivors of childhood cancer in the United States, and most of them receive their medical care from primary care physicians. Adult survivors of childhood cancer are at considerable risk of long-term morbidity and mortality beyond the recurrence of their primary malignancy. Late adverse effects can impair organ function, stunt growth and development, and cause neurocognitive dysfunction and secondary malignancies. To address the need for systematic, comprehensive care of this expanding high-risk patient population, the Children's Oncology Group has developed long-term follow-up guidelines. Proper use of these guidelines will allow primary care physicians to understand a patient's individual risk, provide additional screening as needed, and identify late adverse effects of childhood cancer early. The foundation of the care of an adult survivor of a childhood cancer is a complete, accurate account of the patient's cancer and subsequent therapy in the form of a Summary of Cancer Treatment. A complete Summary of Cancer Treatment allows a primary care physician to use the longterm follow-up guidelines to create an individualized care plan. This article will review the late adverse effects of childhood cancer therapy and the transition of patients from pediatric oncologists to physicians in adulthood, and explain how primary care physicians can use these tools to provide appropriate care to adult survivors of childhood cancer.


T-bet upregulation and subsequent interleukin 12 stimulation are essential for induction of Th1 mediated immunopathology in Crohn's disease.
Many lines of evidence suggest that T helper cell type 1 (Th1) immune responses predominate in Crohn's disease (CD). Recently, a novel transcription factor T-box expressed in T cells (T-bet) has been reported as the master regulator of Th1 development. This study was designed to investigate the role of T-bet and proinflammatory cytokines in Th1 mediated immunopathology in CD. CD4+ lamina propria mononuclear cells (LPMCs) were isolated from surgically resected specimens (CD, n = 10; ulcerative colitis (UC), n = 10; normal controls (NL), n = 5). (1) T-bet expression of CD4+ LPMCs was examined by quantitative real time polymerase chain reaction and western blotting. (2) T-bet expression of LPMCs stimulated by interleukin (IL)-12/IL-18 was analysed by western blotting. (3) Interferon gamma (IFN-gamma) production and T-bet expression of CD4+ peripheral blood mononuclear cells (PBMCs) were examined with or without stimulation by anti-CD3/CD28 monoclonal antibodies and/or IL-12. (1) T-bet expression of CD4+ LPMCs was increased in CD compared with UC and NL. (2) Synergistically, augmentation of IFN-gamma production by IL-12/IL-18 was independent of T-bet expression in LPMCs. (3) T-bet was induced by T cell receptor stimulation in CD4+ PBMCs. T-bet induction correlated with IFN-gamma production and with augmentation of surface expressed IL-12 receptor beta2. T-bet induction by antigenic stimulation and subsequent stimulation by macrophage derived IL-12/IL-18 are important for establishing Th1 mediated immunopathology in CD.


Step-by-step engineered multiparticle entanglement
After quantum particles have interacted, they generally remain in an entangled state and are correlated at a distance by quantum-mechanical links that can be used to transmit and process information in nonclassical ways. This implies programmable sequences of operations to generate and analyze the entanglement of complex systems. We have demonstrated such a procedure for two atoms and a single-photon cavity mode, engineering and analyzing a three-particle entangled state by a succession of controlled steps that address the particles individually. This entangling procedure can, in principle, operate on larger numbers of particles, opening new perspectives for fundamental tests of quantum theory.


Cross-View Retrieval via Probability-Based Semantics-Preserving Hashing.
For efficiently retrieving nearest neighbors from large-scale multiview data, recently hashing methods are widely investigated, which can substantially improve query speeds. In this paper, we propose an effective probability-based semantics-preserving hashing (SePH) method to tackle the problem of cross-view retrieval. Considering the semantic consistency between views, SePH generates one unified hash code for all observed views of any instance. For training, SePH first transforms the given semantic affinities of training data into a probability distribution, and aims to approximate it with another one in Hamming space, via minimizing their Kullback-Leibler divergence. Specifically, the latter probability distribution is derived from all pair-wise Hamming distances between to-be-learnt hash codes of the training data. Then with learnt hash codes, any kind of predictive models like linear ridge regression, logistic regression, or kernel logistic regression, can be learnt as hash functions in each view for projecting the corresponding view-specific features into hash codes. As for out-of-sample extension, given any unseen instance, the learnt hash functions in its observed views can predict view-specific hash codes. Then by deriving or estimating the corresponding output probabilities with respect to the predicted view-specific hash codes, a novel probabilistic approach is further proposed to utilize them for determining a unified hash code. To evaluate the proposed SePH, we conduct extensive experiments on diverse benchmark datasets, and the experimental results demonstrate that SePH is reasonable and effective.


Angiosarcomas of primary gynecologic origin: a clinicopathologic review and quantitative analysis of survival.
Angiosarcomas are aggressive, malignant soft tissue neoplasms of endothelial origin and occur rarely in the female genital tract. There is lack of consensus on risk factors for poor outcome and optimal treatment. To this end, we performed a clinicopathologic review and survival analysis. We report a case of a woman with an angiosarcoma of the vagina. Published English literature was reviewed for angiosarcomas of the vulva, vagina, uterus, and ovary. Survival was evaluated by using Kaplan-Meier analysis and the effect of clinical and demographic variables on survival by using Cox regression analysis. A total of 51 patients were identified with a median age of 47 years (range, 17-87 years). Two of the patients had an angiosarcoma of the vulva; 2 had an angiosarcoma of the vagina; 18 had an angiosarcoma of the uterus, and 29 had an angiosarcoma of the ovary. Five-year overall survival was 27% (SE, 8%). Most patients presented with locoregional disease, having surgery as their primary intervention. Overall, adjuvant therapy significantly improved survival (hazards ratio, 0.17; 95% confidence interval, 0.05-0.59; adjusted for age and tumor size). Adjuvant treatment consisted of radiotherapy for angiosarcomas of the vulva, vagina, and uterus and chemotherapy for ovarian angiosarcomas. Subgroup analysis of the female genital tract site was hampered by the small number of cases. This review supports the use of surgical and adjuvant radiotherapy for angiosarcomas of the vulva, vagina, and uterus. Cytoreductive surgery and adjuvant chemotherapy remain the primary treatment of angiosarcomas of the ovary.


Collaborative care management of major depression among low-income, predominantly Hispanic subjects with diabetes: a randomized controlled trial.
To determine whether evidence-based socioculturally adapted collaborative depression care improves receipt of depression care and depression and diabetes outcomes in low-income Hispanic subjects. This was a randomized controlled trial of 387 diabetic patients (96.5% Hispanic) with clinically significant depression recruited from two public safety-net clinics from August 2005 to July 2007 and followed over 18 months. Intervention (INT group) included problem-solving therapy and/or antidepressant medication based on a stepped-care algorithm; first-line treatment choice; telephone treatment response, adherence, and relapse prevention follow-up over 12 months; plus systems navigation assistance. Enhanced usual care (EUC group) included standard clinic care plus patient receipt of depression educational pamphlets and a community resource list. INT patients had significantly greater depression improvement (> or =50% reduction in Symptom Checklist-20 depression score from baseline; 57, 62, and 62% vs. the EUC group's 36, 42, and 44% at 6, 12, and 18 months, respectively; odds ratio 2.46-2.57; P < 0.001). Mixed-effects linear regression models showed a significant study group-by-time interaction over 18 months in diabetes symptoms; anxiety; Medical Outcomes Study Short-Form Health Survey (SF-12) emotional, physical, and pain-related functioning; Sheehan disability; financial situation; and number of social stressors (P = 0.04 for disability and SF-12 physical functioning, P < 0.001 for all others) but no study group-by-time interaction in A1C, diabetes complications, self-care management, or BMI. Socioculturally adapted collaborative depression care improved depression, functional outcomes, and receipt of depression treatment in predominantly Hispanic patients in safety-net clinics.


Culture-dependent and culture-independent methods reveal diverse methylotrophic communities in terrestrial environments.
One-carbon compounds such as methanol, dimethylsulfide (DMS) and dimethylsulfoxide (DMSO) are significant intermediates in biogeochemical cycles. They are suggested to affect atmospheric chemistry and global climate. Methylotrophic microorganisms are considered as a significant sink for these compounds; therefore, we analyzed the diversity of terrestrial bacteria that utilize methanol, DMS and DMSO as carbon and energy source using culture-dependent and culture-independent methods. The effect of habitat type on the methylotrophic community structure was also investigated in rhizosphere and bulk soil. While thirteen strains affiliated to the genera Hyphomicrobium, Methylobacterium, Pseudomonas, Hydrogenophaga, Rhodococcus, Flavobacterium and Variovorax were isolated, denaturing gradient gel electrophoresis revealed the dominance of Thiobacillus, Rhodococcus, Flavobacterium and Bacteroidetes species. Furthermore, methylotrophic communities that degrade methanol or DMS are not shaped by terrestrial habitat type. Rhizosphere and soil samples showed dominance of Methylophilus spp. and Methylovorus spp. for methanol enrichments; Cytophaga spp., Pseudomonas tremae and Thiobacillus thioparus for DMS enrichments.


Distinct plasma profile of polar neutral amino acids, leucine, and glutamate in children with Autism Spectrum Disorders.
The goal of this investigation was to examine plasma amino acid (AA) levels in children with Autism Spectrum Disorders (ASD, N = 27) and neuro-typically developing controls (N = 20). We observed reduced plasma levels of most polar neutral AA and leucine in children with ASD. This AA profile conferred significant post hoc power for discriminating children with ASD from healthy children. Furthermore, statistical correlations suggested the lack of a typical decrease of glutamate and aspartate with age, and a non-typical increase of isoleucine and lysine with age in the ASD group. Findings from this limited prospective study warrant further examination of plasma AA levels in larger cross-sectional and longitudinal cohorts to adequately assess for relationships with developmental and clinical features of ASD.


Homologies between different procaryotic DNA-binding regulatory proteins and between their sites of action.
Comparison of the amino acid sequences of 13 procaryotic regulatory proteins, including the products of genes crp (catabolite activator protein; CAP), lacI, galR , lexA, lysR, araC, trpR, and tnpR of Escherichia coli, of genes cI, cII and cro of phage lambda, cro of phage 434, and c2 of phage P22, has revealed two regions of homology. The sites of action of these proteins also share common features in their DNA sequence. Taking into account the models proposed for the lambda repressors, cro and cI, and for CAP, a general type of DNA-protein interaction is suggested.


Genetic and Morphological Variation of the Forkbeard, Phycis phycis (Pisces, Phycidae): Evidence of Panmixia and Recent Population Expansion along Its Distribution Area.
The knowledge of population structure of a species is essential to effectively assess and manage fisheries. In the present study, genetics, by mitochondrial DNA cytochrome b sequence analysis, and body geometric morphometrics were used to evaluate the existence of distinct populations of the forkbeard (Phycis phycis), an important commercial species in several European countries, especially Portugal and Spain. For geometric morphometric analysis, specimens were collected in the Northeast Atlantic Ocean-Azores, Madeira and mainland Portugal, and for genetic analysis, these samples were complemented with samples collected in the Mediterranean Sea-Spain, Italy and Croatia, in order to cover the entire distribution area of the species. Body shape of the forkbeard from the Northeast Atlantic was found to be highly variable. This variation was probably associated with different environmental factors between the study areas. Despite morphological variation, a low genetic differentiation between samples from different areas was found, most likely due to gene flow that occurred in the past or with the demographic history of the species. Moreover, the presence of unique haplotypes in the Northeast Atlantic and in the Mediterranean suggests that recent gene flow between populations from these areas should be limited. Altogether, a high haplotype diversity, a low nucleotide diversity, a "star-like" network and the results of the mismatch distribution, indicate a possible signature of recent population expansions, which probably started during the end of the Last Glacial Maximum and led to the colonization of the Northeast Atlantic and the Mediterranean.


Centers for medicare and medicaid services hospital-acquired conditions policy for central line-associated bloodstream infection (CLABSI) and catheter-associated urinary tract infection (CAUTI) shows minimal impact on hospital reimbursement.
In 2008, the Centers for Medicare and Medicaid Services (CMS) stopped reimbursing for hospital-acquired conditions (HACs) not present on admission (POA). We sought to understand why this policy did not impact central line-associated bloodstream infection (CLABSI) and catheter-associated urinary tract infection (CAUTI) trends. Retrospective cohort study. Acute-care hospitals in the United States.ParticipantsFee-for-service Medicare patients discharged January 1, 2007, through December 31, 2011. Using inpatient Medicare claims data, we analyzed billing practices before and after the HAC policy was implemented, including the use and POA designation of codes for CLABSI or CAUTI. For the 3-year period following policy implementation, we determined the impact on diagnosis-related groups (DRG) determining reimbursement as well as hospital characteristics associated with the reimbursement impact. During the study period, 65,205,607 Medicare fee-for-service hospitalizations occurred at 3,291 acute-care, nonfederal US hospitals. Based on coding, CLABSI and CAUTI affected 0.23% and 0.06% of these hospitalizations, respectively. In addition, following the HAC policy, 82% of the CLABSI codes and 91% of the CAUTI codes were marked POA, which represented a large increase in the use of this designation. Finally, for the small numbers of CLABSI and CAUTI coded as not POA, financial impacts were detected on only 0.4% of the hospitalizations with a CLABSI code and 5.7% with a CAUTI code. Part of the reason the HAC policy did not have its intended impact is that billing codes for CLABSI and CAUTI were rarely used, were commonly listed as POA in the postpolicy period, and infrequently impacted hospital reimbursement.


Short malunions of the clavicle: an anatomic and functional study.
The anatomic and functional effects of clavicle malunion, defined as 15 mm of shortening, were evaluated in 10 subjects. Bilateral shoulder computed tomography with 3-dimensional reconstructions was performed. A self-administered questionnaire was completed, and biomechanical testing of subjects was undertaken comparing strength and velocity of movement between shoulders. Mean shoulder scores and visual analog global assessments of shoulder function were significantly worse in the injured shoulder. Biomechanical assessment recorded a reduction in muscular strength for adduction, extension, and internal rotation of the humerus as well as a reduced peak abduction velocity in the injured shoulder (P < .05). Computed tomography assessment showed that clavicular shortening produced increased upward angulation of the clavicle at the sternoclavicular joint (P < .005) and increased anterior scapular version (P < .05). These changes in static anatomic relationships are possible mechanisms limiting shoulder function after short clavicle malunions. Consideration should be given to the prevention of shortening by open reduction and internal fixation, especially in the young, active age group.


Toca-1 mediates Cdc42-dependent actin nucleation by activating the N-WASP-WIP complex.
An important signaling pathway to the actin cytoskeleton links the Rho family GTPase Cdc42 to the actin-nucleating Arp2/3 complex through N-WASP. Nevertheless, these previously identified components are not sufficient to mediate Cdc42-induced actin polymerization in a physiological context. In this paper, we describe the biochemical purification of Toca-1 (transducer of Cdc42-dependent actin assembly) as an essential component of the Cdc42 pathway. Toca-1 binds both N-WASP and Cdc42 and is a member of the evolutionarily conserved PCH protein family. Toca-1 promotes actin nucleation by activating the N-WASP-WIP/CR16 complex, the predominant form of N-WASP in cells. Thus, the cooperative actions of two distinct Cdc42 effectors, the N-WASP-WIP complex and Toca-1, are required for Cdc42-induced actin assembly. These findings represent a significantly revised view of Cdc42-signaling and shed light on the pathogenesis of Wiskott-Aldrich syndrome.


The nature of non-appendicular anterior paired projections in Palaeozoic total-group Euarthropoda.
Recent studies have clarified the segmental organization of appendicular and exoskeletal structures in the anterior region of Cambrian stem-group Euarthropoda, and thus led to better understanding of the deep evolutionary origins of the head region in this successful animal group. However, there are aspects of the anterior organization of Palaeozoic euarthropods that remain problematic, such as the morphological identity and significance of minute limb-like projections on the anterior region in stem and crown-group representatives. Here, we draw attention to topological and morphological similarities between the frontal filaments of extant Crustacea and the embryonic frontal processes of Onychophora, and distinctive anterior paired projections observed in several extinct total-group Euarthropoda. Anterior paired projections are redescribed in temporally and phylogenetically distant fossil taxa, including the gilled lobopodians Kerygmachela kierkegaardi and Pambdelurion whittingtoni, the bivalved stem-euarthropod Canadaspis perfecta, the larval pycnogonid Cambropycnogon klausmuelleri, and the mandibulate Tanazios dokeron. Developmental data supporting the homology of the 'primary antennae' of Onychophora, the 'frontal appendages' of lower-stem Euarthropoda, and the hypostome/labrum complex of Deuteropoda, argue against the morphological identity of the anterior paired projections of extant and extinct panarthropods as a pair of pre-ocular appendages. Instead, we regard the paired projections of fossil total-group euarthropods as non-appendicular evaginations with a likely protocerebral segmental association, and a possible sensorial function. The widespread occurrence of pre-ocular paired projections among extant and extinct taxa suggests their potential homology as fundamentally ancestral features of the anterior body organization in Panarthropoda. Non-appendicular paired projections with a sensorial function may reflect a critical--yet previously overlooked--component of the panarthropod ground pattern.


The MDR1 (ABCB1) gene polymorphism and its clinical implications.
There has been an increasing appreciation of the role of drug transporters in the pharmacokinetic and pharmacodynamic profiles of certain drugs. Among various drug transporters, P-glycoprotein, the MDR1 gene product, is one of the best studied and characterised. P-glycoprotein is expressed in normal human tissues such as liver, kidney, intestine and the endothelial cells of the blood-brain barrier. Apical (or luminal) expression of P-glycoprotein in these tissues results in reduced drug absorption from the gastrointestinal tract, enhanced drug elimination into bile and urine, and impeded entry of certain drugs into the central nervous system. The clinical relevance of P-glycoprotein depends on the localisation in human tissues (i.e. vectorial or directional movement), the therapeutic index of the substrate drug and the inherent inter- and intra-individual variability. With regard to the variability, polymorphisms of the MDR1 gene have recently been reported to be associated with alterations in disposition kinetics and interaction profiles of clinically useful drugs, including digoxin, fexofenadine, ciclosporin and talinolol. In addition, polymorphism may play a role in patients who do not respond to drug treatment. Moreover, P-glycoprotein is an important prognostic factor in malignant diseases, such as tumours of the gastrointestinal tract.A growing number of preclinical and clinical studies have demonstrated that polymorphism of the MDR1 gene may be a factor in the overall outcome of pharmacotherapy for numerous diseases. We believe that further understanding the physiology and biochemistry of P-glycoprotein with respect to its genetic variations will be important to establish individualised pharmacotherapy with various clinically used drugs.


Differential effects of streptozotocin-induced diabetes on cardiac lipoprotein lipase activity.
Lipoprotein lipase (LPL) is an endothelial-bound enzyme that is rate determining for the clearance of triacylglycerol-rich lipoproteins. Previous studies using rats with streptozotocin (STZ)-induced diabetes have reported inconsistent effects on cardiac LPL activity or immunoreactive protein. To examine the contribution of the severity and duration of diabetes on cellular and heparin-releasable cardiac LPL activity, Wistar rats were administered a high (100 mg/kg; D100) or moderate (55 mg/kg; D55) dose of STZ, and LPL activity was examined at various times after diabetes induction. Heparin perfusion of the isolated Langendorff control heart induced the release of LPL activity as an initial fast phase followed by a slow phase of release. With increasing age, the second phase of LPL release became more pronounced. Severe STZ-induced diabetes reduced heparin-releasable LPL activity by 1 week in the D100 rats. However, in D55 rat hearts, peak heparin-releasable LPL activity was higher than that in control animals at 2 and 12 weeks after STZ injection, with a complete absence of the delayed phase at 12 weeks. The elevated heparin-releasable LPL peak could not be explained by an enhanced LPL synthesis because both cellular and surface-bound LPL activities in myocytes from D55 rats were low, relative to control. Chronic (12-day) insulin treatment of D55 rats prevented the rise in heparin-releasable LDL and the reduction in cell-associated LPL activity. Moreover, acute (90-min) treatment of D55 rats with rapid-acting insulin also reduced the heparin-releasable LPL activity to normal levels, although it had no effect on the low cellular LPL activity. When the heparin-releasable LPL pool was allowed to recover for 1 h after removal of the enzyme, D55 rat hearts continued to demonstrate a higher peak LPL activity after a second heparin perfusion. These studies demonstrate that in moderate but not severe diabetes, there is an augmented peak heparin-releasable LPL activity. Whether or not this enhanced heparin-releasable LPL activity has a pathological role in the diabetic heart has yet to be determined.


A method for describing and evaluating naturopathic whole practice.
Even though complementary and alternative medicine (CAM) is generally practiced as distinct systems of medicine, almost all CAM research has focused on single therapies. In order to more adequately evaluate the effectiveness of these medical systems, studies that evaluate the outcome of intact whole systems are needed. One challenge lies in defining the whole medical system (and any medical system it is compared to) in a way that ensures treatment fidelity. This paper presents a proposed method to measure treatment fidelity (treatment criteria) in studies of the naturopathic medical system. Illustrative example of the theory-based development and post-hoc "testing" of treatment criteria against an existing database of actual treatments prescribed by a random sample of naturopathic physicians. Treatment criteria for 3 conditions--menopausal symptoms, bowel dysfunction, and fatigue/fibromyalgia--and their comparison to actual treatments prescribed. A set of meaningful, measurable treatment criteria based on the naturopathic practice principles were defined that could have generated the majority (82%-93%) of treatment prescriptions given at visits for these conditions. Several of the treatment criteria components are common across the 3 conditions studied, and might be appropriate for all visits to doctors of naturopathy (NDs). Others are specific to each condition. In addition to ensuring model validity, these criteria help identify critical components of care, enable study replication, provide a measure of quality of care, and are one step toward allowing CAM to be studied as it is generally practiced-as distinct systems of medicine. Work was performed at Bastyr University and the University of Arizona.


Risk of postpartum episodes in women with bipolar disorder after lamotrigine or lithium use during pregnancy: A population-based cohort study.
Women with bipolar disorder are at high risk for relapse/recurrence postpartum. Among all mood stabilizers, lithium has the largest evidence base for efficacy in the peripartum period, but lamotrigine is increasingly prescribed for bipolar spectrum disorders during pregnancy. The aim of this study was to investigate whether lamotrigine use during pregnancy is as effective as lithium in the prevention of severe episodes postpartum. Danish national registries were used to identify pregnancies of women with a diagnosis of bipolar spectrum disorders at the time of conception who used lamotrigine or lithium during pregnancy. We compared the risk of inpatient psychiatric admission within three months postpartum between women who used lamotrigine (N=55) versus lithium (N=59) during pregnancy. A logistic regression model was used to calculate crude and adjusted odds ratios. We did not find a significant difference in the risk of postpartum psychiatric admission between women who used lamotrigine versus lithium during pregnancy (7.3% versus 15.3% respectively, adjusted OR 0.83; 95% CI 0.22-3.14). We adjusted for year of delivery, parity, previous admissions and antidepressant/benzodiazepine use during pregnancy. Other variables did not differ substantially between groups. We used an observational design and therefore patients were not randomized to lamotrigine or lithium. The study has a small sample size. Lamotrigine was not inferior to lithium in the prevention of severe postpartum episodes. Our findings suggest lamotrigine could be a reasonable alternative treatment option for bipolar disorder during pregnancy in patients with vulnerability for depression and may prevent severe episodes postpartum.


Rapid, precise, and reliable measurement of delay discounting using a Bayesian learning algorithm.
Machine learning has the potential to facilitate the development of computational methods that improve the measurement of cognitive and mental functioning. In three populations (college students, patients with a substance use disorder, and Amazon Mechanical Turk workers), we evaluated one such method, Bayesian adaptive design optimization (ADO), in the area of delay discounting by comparing its test-retest reliability, precision, and efficiency with that of a conventional staircase method. In all three populations tested, the results showed that ADO led to 0.95 or higher test-retest reliability of the discounting rate within 10-20 trials (under 1-2 min of testing), captured approximately 10% more variance in test-retest reliability, was 3-5 times more precise, and was 3-8 times more efficient than the staircase method. The ADO methodology provides efficient and precise protocols for measuring individual differences in delay discounting.


Single nucleotide polymorphism (SNP) discovery in mammals: a targeted-gene approach.
Single nucleotide polymorphisms (SNPs) have rarely been exploited in nonhuman and nonmodel organism genetic studies. This is due partly to difficulties in finding SNPs in species where little DNA sequence data exist, as well as to a lack of robust and inexpensive genotyping methods. We have explored one SNP discovery method for molecular ecology, evolution, and conservation studies to evaluate the method and its limitations for population genetics in mammals. We made use of 'CATS' (or 'EPIC') primers to screen for novel SNPs in mammals. Most of these primer sets were designed from primates and/or rodents, for amplifying intron regions from conserved genes. We have screened 202 loci in 16 representatives of the major mammalian clades. Polymerase chain reaction (PCR) success correlated with phylogenetic distance from the human and mouse sequences used to design most primers; for example, specific PCR products from primates and the mouse amplified the most consistently and the marsupial and armadillo amplifications were least successful. Approximately 24% (opossum) to 65% (chimpanzee) of primers produced usable PCR product(s) in the mammals tested. Products produced generally high but variable levels of readable sequence and similarity to the expected genes. In a preliminary screen of chimpanzee DNA, 12 SNPs were identified from six (of 11) sequenced regions, yielding a SNP on average every 400 base pairs (bp). Given the progress in genome sequencing, and the large numbers of CATS-like primers published to date, this approach may yield sufficient SNPs per species for population and conservation genetic studies in nonmodel mammals and other organisms.


Psychiatrist's adherence: a new factor in relapse prevention of schizophrenia. A randomized controlled study on relapse control through telemedicine system.
Exposure to psychotic states has detrimental effects on the long-term outcome of schizophrenia and brain integrity. Therefore, improving relapse prevention is a key component of long-term management of schizophrenia. Previous studies using continuous monitoring of an individual's early signs of relapse and adopting preventative pharmacological interventions, when early signs are detected, showed promising clinical results in terms of relapse risk reduction. This 18-month multi-centre parallel randomized controlled, open label, trial with telemedicine relapse prevention programme ITAREPS failed to show superiority of maintenance plus prodrome-based targeted medication strategy over treatment as usual. The study, marked by low investigator's adherence, confirmed that absence of pharmacological intervention at early stage of prodrome, critically influenced the risk of relapse. This and previous randomized controlled trials with telemedicine programme ITAREPS suggested that substantial improvement in relapse prevention in schizophrenia is likely to be unattainable under current clinical settings. Future preventive strategies in schizophrenia would require rapid pharmacological intervention upon occurrence of subclinical prodromal symptoms that are undetectable under conventional outpatient practice. Studies with ITAREPS suggested that integration of telemedicine relapse prevention systems and visiting nurse service might together represent practical solution capable to address those requirements. The Information Technology Aided Relapse Prevention Programme in Schizophrenia (ITAREPS) presents a telemedicine solution for weekly monitoring and management of schizophrenia. This study aims to evaluate the effectiveness of the programme in reducing the number of hospitalizations during the 18-month multi-centre parallel randomized controlled, open label, trial. Outpatients with schizophrenia or schizoaffective disorder were randomized to the active (n = 74) or control group (n = 72). In the active arm, investigators increased the antipsychotic dose upon occurrence of prodrome announced by the system. Intention-to-treat analysis showed no between-group difference in the hospitalization-free survival rate [Kaplan-Meier method; hazard ratio (HR) = 1.21, 95% confidence interval (CI): 0.56-2.61, P = 0.6). In a post hoc multivariate Cox proportional hazards model, out of 13 potential predictors, only ITAREPS-related variables (number of alerts without pharmacological intervention/HR = 1.38, P = 0.042/ and patient non-adherence with ITAREPS /HR = 1.08, P = 0.009/) increased the risk of hospitalization. In this trial ITAREPS was not effective. The results in context with previous ITAREPS studies suggest non-adherence of both psychiatrists and patients as the main reasons for the failure of this preventive strategy. Tertiary prevention in schizophrenia have to be regarded a major challenge, warranting the need for implementation of strategies with more active participation of both patient and treating psychiatrist.


Beyond an Updated Graded Prognostic Assessment (Breast GPA): A Prognostic Index and Trends in Treatment and Survival in Breast Cancer Brain Metastases From 1985 to Today.
Brain metastases are a common sequelae of breast cancer. Survival varies widely based on diagnosis-specific prognostic factors (PF). We previously published a prognostic index (Graded Prognostic Assessment [GPA]) for patients with breast cancer with brain metastases (BCBM), based on cohort A (1985-2007, n = 642), then updated it, reporting the effect of tumor subtype in cohort B (1993-2010, n = 400). The purpose of this study is to update the Breast GPA with a larger contemporary cohort (C) and compare treatment and survival across the 3 cohorts. A multi-institutional (19), multinational (3), retrospective database of 2473 patients with breast cancer with newly diagnosed brain metastases (BCBM) diagnosed from January 1, 2006, to December 31, 2017, was created and compared with prior cohorts. Associations of PF and treatment with survival were analyzed. Kaplan-Meier survival estimates were compared with log-rank tests. PF were weighted and the Breast GPA was updated such that a GPA of 0 and 4.0 correlate with the worst and best prognoses, respectively. Median survival (MS) for cohorts A, B, and C improved over time (from 11, to 14 to 16 months, respectively; P < .01), despite the subtype distribution becoming less favorable. PF significant for survival were tumor subtype, Karnofsky Performance Status, age, number of BCBMs, and extracranial metastases (all P < .01). MS for GPA 0 to 1.0, 1.5-2.0, 2.5-3.0, and 3.5-4.0 was 6, 13, 24, and 36 months, respectively. Between cohorts B and C, the proportion of human epidermal receptor 2 + subtype decreased from 31% to 18% (P < .01) and MS in this subtype increased from 18 to 25 months (P < .01). MS has improved modestly but varies widely by diagnosis-specific PF. New PF are identified and incorporated into an updated Breast GPA (free online calculator available at brainmetgpa.com). The Breast GPA facilitates clinical decision-making and will be useful for stratification of future clinical trials. Furthermore, these data suggest human epidermal receptor 2-targeted therapies improve clinical outcomes in some patients with BCBM.


Developmental exposure to a complex PAH mixture causes persistent behavioral effects in naive Fundulus heteroclitus (killifish) but not in a population of PAH-adapted killifish.
Acute exposures to some individual polycyclic aromatic hydrocarbons (PAHs) and complex PAH mixtures are known to cause cardiac malformations and edema in the developing fish embryo. However, the heart is not the only organ impacted by developmental PAH exposure. The developing brain is also affected, resulting in lasting behavioral dysfunction. While acute exposures to some PAHs are teratogenically lethal in fish, little is known about the later life consequences of early life, lower dose subteratogenic PAH exposures. We sought to determine and characterize the long-term behavioral consequences of subteratogenic developmental PAH mixture exposure in both naive killifish and PAH-adapted killifish using sediment pore water derived from the Atlantic Wood Industries Superfund Site. Killifish offspring were embryonically treated with two low-level PAH mixture dilutions of Elizabeth River sediment extract (ERSE) (TPAH 5.04 μg/L and 50.4 μg/L) at 24h post fertilization. Following exposure, killifish were raised to larval, juvenile, and adult life stages and subjected to a series of behavioral tests including: a locomotor activity test (4 days post-hatch), a sensorimotor response tap/habituation test (3 months post hatch), and a novel tank diving and exploration test (3months post hatch). Killifish were also monitored for survival at 1, 2, and 5 months over 5-month rearing period. Developmental PAH exposure caused short-term as well as persistent behavioral impairments in naive killifish. In contrast, the PAH-adapted killifish did not show behavioral alterations following PAH exposure. PAH mixture exposure caused increased mortality in reference killifish over time; yet, the PAH-adapted killifish, while demonstrating long-term rearing mortality, had no significant changes in mortality associated with ERSE exposure. This study demonstrated that early embryonic exposure to PAH-contaminated sediment pore water caused long-term locomotor and behavioral alterations in killifish, and that locomotor alterations could be observed in early larval stages. Additionally, our study highlights the resistance to behavioral alterations caused by low-level PAH mixture exposure in the adapted killifish population. Furthermore, this is the first longitudinal behavioral study to use killifish, an environmentally important estuarine teleost fish, and this testing framework can be used for future contaminant assessment.


Multi-step self-guided pathways for shape-changing metamaterials.
Multi-step pathways-which consist of a sequence of reconfigurations of a structure-are central to the functionality of various natural and artificial systems. Such pathways execute autonomously in self-guided processes such as protein folding1 and self-assembly2-5, but have previously required external control to execute in macroscale mechanical systems, provided by, for example, actuators in robotics6-9 or manual folding in origami8,10-12. Here we demonstrate shape-changing, macroscale mechanical metamaterials that undergo self-guided, multi-step reconfiguration in response to global uniform compression. We avoid the need for external control by using metamaterials that are made purely of passive components. The design of the metamaterials combines nonlinear mechanical elements with a multimodal architecture that enables a sequence of topological reconfigurations caused by the formation of internal self-contacts between the elements of the metamaterial. We realize the metamaterials by using computer-controlled water-jet cutting of flexible materials, and show that the multi-step pathway and final configuration can be controlled by rational design of the nonlinear mechanical elements. We also demonstrate that the self-contacts suppress errors in the pathway. Finally, we create hierarchical architectures to extend the number of distinct reconfiguration steps. Our work establishes general principles for designing mechanical pathways, opening up new avenues for self-folding media11,12, pluripotent materials9,13 and pliable devices14 in areas such as stretchable electronics and soft robotics15.


Aberrant epithelial-mesenchymal Hedgehog signaling characterizes Barrett's metaplasia.
The molecular mechanism underlying epithelial metaplasia in Barrett's esophagus remains unknown. Recognizing that Hedgehog signaling is required for early esophageal development, we sought to determine if the Hedgehog pathway is reactivated in Barrett's esophagus, and if genes downstream of the pathway could promote columnar differentiation of esophageal epithelium. Immunohistochemistry, immunofluorescence, and quantitative real-time polymerase chain reaction were used to analyze clinical specimens, human esophageal cell lines, and mouse esophagi. Human esophageal squamous epithelial (HET-1A) and adenocarcinoma (OE33) cells were subjected to acid treatment and used in transfection experiments. Swiss Webster mice were used in a surgical model of bile reflux injury. An in vivo transplant culture system was created using esophageal epithelium from Sonic hedgehog transgenic mice. Marked up-regulation of Hedgehog ligand expression, which can be induced by acid or bile exposure, occurs frequently in Barrett's epithelium and is associated with stromal expression of the Hedgehog target genes PTCH1 and BMP4. BMP4 signaling induces expression of SOX9, an intestinal crypt transcription factor, which is highly expressed in Barrett's epithelium. We further show that expression of Deleted in Malignant Brain Tumors 1, the human homologue of the columnar cell factor Hensin, occurs in Barrett's epithelium and is induced by SOX9. Finally, transgenic expression of Sonic hedgehog in mouse esophageal epithelium induces expression of stromal Bmp4, epithelial Sox9, and columnar cytokeratins. Epithelial Hedgehog ligand expression may contribute to the initiation of Barrett's esophagus through induction of stromal BMP4, which triggers reprogramming of esophageal epithelium in favor of a columnar phenotype.


MAFL experiment: development of photonic devices for a space-based multiaperture fiber-linked interferometer.
We present a three-telescope space-based interferometer prototype dedicated to high-resolution imaging. This project, named multiaperture fiber-linked interferometer (MAFL), was founded by the European Space Agency. The aim of the MAFL project is to propose, design, and implement for the first time to the best of our knowledge all the optical functions required for the global instrument on the same integrated optics (IO) component for controlling a three-arm interferometer and to obtain reliable science data. The coherent transport from telescopes to the IO component is achieved by means of highly birefringent optical fiber. The laboratory bench is presented, and the results are reported allowing us to validate the optical potentiality of the IO component in this frame. The validation measurements consist of the throughput of this optical device, the performances of metrological servoloop, and the instrumental contrasts and phase closure of the science fringes.


Proliferating cell nuclear antigen in oesophageal diseases; correlation with transforming growth factor alpha expression.
This study was designed to correlate mucosal proliferation in Barrett's oesophagus with expression of a growth promoting peptide, transforming growth factor alpha (TGF alpha). Oesophageal mucosa was studied from 50 patients with oesophageal disease who had been treated by oesophagectomy. Histological analysis showed a range of oesophageal pathology - 18 patients had gastric type Barrett's mucosa, 18 had intestinal type Barrett's mucosa, and 14 had oesophageal adenocarcinomas. Sections were stained immunohistochemically for proliferating cell nuclear antigen (PCNA) (an index of cellular proliferation) and TGF alpha. PCNA immunostaining was seen mainly in the basal cells of the neck/foveolar epithelial compartment of the glands in Barrett's oesophagus. However, in mucosa with high grade dysplasia, the proliferative compartment extended upwards into the superficial layers of the glands. At least 2000 cells were counted in each patient to determine the proportion with PCNA immunoreactivity (PCNA labelling index). The labelling index was highest in adenocarcinoma (25%) and in Barrett's intestinal type mucosa with high grade dysplasia (26%) compared with intestinal type mucosa with no significant dysplasia (20%) and Barrett's gastric type mucosa (12%). There was a significant positive correlation between PCNA labelling indices and TGF alpha expression in Barrett's mucosa (p less than 0.01). In glands showing high grade dysplasia, TGF alpha immunoreactivity was seen in the same regions of the glands as PCNA immunoreactivity, indicating the possibility of involvement of TGF alpha in (pre) neoplastic proliferation in Barrett's oesophagus.


Calcifediol versus vitamin D3 effects on gait speed and trunk sway in young postmenopausal women: a double-blind randomized controlled trial.
In this double-blind RCT, 4-month treatment with calcifediol compared with vitamin D3 improved gait speed by 18% among young postmenopausal women. Consistently, change in 25(OH)D blood levels over time were significantly correlated with improvement in gait speed in these women. No effect could be demonstrated for trunk sway. The aim of this study is to test the effect of calcifediol compared with vitamin D3 on gait speed and trunk sway. Twenty healthy postmenopausal women with an average 25(OH)D level of 13.2 ng/ml (SD = ±3.9) and a mean age of 61.5 years (SD = ±7.2) were randomized to either 20 μg of calcifediol or 20 μg (800 IU) of vitamin D3 per day in a double-blind manner. At baseline and at 4 months of follow-up, the same physiotherapist blinded to treatment allocation tested 8-m gait speed and a body sway test battery (Sway star pitch and roll angle plus velocity while walking 8 m, and standing on both legs on a hard and soft surface). All analyses adjusted for baseline measurement, age, and body mass index. Mean 25(OH)D levels increased to 69.3 ng/ml (SD = ±9.5) in the calcifediol group and to 30.5 ng/ml (SD = ±5.0) in the vitamin D3 group (p < 0.0001). Women receiving calcifediol compared with vitamin D3 had an 18% greater improvement in gait speed at 4-month follow-up (p = 0.046) adjusting for baseline gait speed, age, and body mass index. Also, change in gait speed was significantly correlated with change in serum 25(OH)D concentrations (r = 0.5; p = 0.04). Across three tests of trunk sway, there were no consistent differences between groups and no significant correlation between change in 25(OH)D serum concentrations and change in trunk sway. Calcifediol improved gait speed in early postmenopausal women compared with vitamin D3 and change in 25(OH)D level was moderately correlated with improvement in gait speed. A benefit on trunk sway could not be demonstrated.


[Social and familial determinants of anxiety and depressive symptoms in middle to low income women].
Among women, risk factors for mental disorders are violence, economical inequities and disadvantages and discrimination. To determine the prevalence of anxiety and depressive symptoms in women and to determine factors associated with them. A cross sectional study of a probabilistic sample of 422 women aged 34 years as a mean, living in a medium to low income neighborhood in Temuco, Chile. Mental health was assessed with the self reporting questionnaire (SRQ20), partner violence was evaluated using a 12 item scale and family conflict and violence was evaluated using the Conflicts Tactics scale. A logistic repression analysis was used to identify factors associated with mental health alterations. Forty one percent of women had anxiety or depressive symptoms. The main associated factors with these symptoms were severe physical violence against children (odds ratio (OR)=14.3), sexual violence against women (OR=9.7), self perception of health (OR=4.5), alcohol abuse (OR=4.4), psychological violence during childhood (OR=3.2) and lack of family support network (OR=2.7). Unemployment and lack of a support network of relatives had an OR 3.3 for mental health alterations. The OR for psychological violence plus sexual violence was 18.5. The figure for psychological violence plus sexual violence plus a history of parental violence during childhood was 26.5. Forty one percent of this group of women had mental health alterations. There is a strong association between these alterations, family violence and socioeconomic factors.


Monitoring the production of inclusion bodies during fermentation and enzyme-linked immunosorbent assay analysis of intact inclusion bodies using cryogel minicolumn plates.
A novel minicolumn chromatographic method to monitor the production of inclusion bodies during fermentation and an enzyme-linked immunosorbent assay (ELISA) system allowing direct analysis of the particles with surface-displayed antigens are described. A 33-kDa protein containing 306 amino acids with three sulfur bridges produced as inclusion bodies was labeled with polyclonal antibodies against 15 amino acid (anti-A15) and 17 amino acid (anti-B17) residues at the N- and C-terminal ends of the protein, respectively. Labeled particles were bound to macroporous monolithic protein A-cryogel adsorbents inserted into the open-ended wells of a 96-well plate (referred to as protein A-cryogel minicolumn plate). The concept behind this application is that the binding degree of inclusion bodies from lysed fermentation broth to the cryogel minicolumns increases with an increase in their concentration during fermentation. The technique allowed us to monitor the increase in the production levels of the inclusion bodies as the fermentation process progressed. The system also has a built-in quality parameter to ensure that the target protein has been fully expressed. Alternatively, inclusion bodies immobilized on phenyl-cryogel minicolumn plate were used in indirect ELISA based on anti-A15 and anti-B17 antibodies against terminal amino acid residues displayed on the surface of inclusion bodies. Drainage-protected properties of the cryogel minicolumns allow performance of successive reactions with tested immunoglobulin G (IgG) samples and enzyme-conjugated secondary IgG and of enzymatic reaction within the adsorbent.


Transmission line model of substrate effects on capacitive mesh couplers.
To use a transmission line model to calculate the optical properties of a thin metal mesh on a dielectric substrate, account must be taken not only of the different propagation conditions within the substrate and of Fabry-Perot resonances due to reflections at the second surface, but also of the effect of the dielectric on the capacitive component of the equivalent reactance of the mesh. Only when this effect is accounted for, which can be done using a simple formula based on Babinet's principle, is good agreement obtained with experimental measurements.


Development of a real-time PCR method for quantification of the three genera Dehalobacter, Dehalococcoides, and Desulfitobacterium in microbial communities.
We developed standard curves based on plasmids containing a 16S rRNA gene of a member of one of the three genera Dehalobacter, Desulfitobacterium, and Dehalococcoides. A large difference in amplification efficiency between the standard curves was observed ranging from 1.5 to 2.0. The total eubacterial 16S rRNA gene copy number determined in a sample DNA by using eubacterial primers and the three standard curves led to differences in the estimated copy numbers of a factor up to 73. However, the amplification efficiencies for one specific standard curve were the same independent of the PCR primer pair used. This allowed the determination of the abundance of a population expressed as fractional number, hence, the percentage of genus-specific copy numbers within the total eubacterial 16S rRNA gene copy numbers. Determination of the fractional numbers in DNA mixtures of known composition showed the accuracy of this approach. The average difference in threshold value between two 10-fold dilutions of DNA of pure cultures, mixtures thereof and of environmental samples was -3.45+/-0.34, corresponding to an average almost optimal amplification efficiency of 1.95. This indicated that the low amplification efficiency of certain standard curves seemed to be mainly a problem of the plasmid DNA used and not of the 16S rRNA gene of the target genera.


The association between type 2 diabetes mellitus, hip fracture, and post-hip fracture mortality: a multi-state cohort analysis.
Type 2 diabetes mellitus (T2DM) is associated with an excess risk of fractures and overall mortality. This study compared hip fracture and post-hip fracture mortality in T2DM and non-diabetic subjects. The salient findings are that subjects in T2DM are at higher risk of dying after suffering a hip fracture. Previous research suggests that individuals with T2DM are at an excess risk of both fractures and overall mortality, but their combined effect is unknown. Using multi-state cohort analyses, we estimate the association between T2DM and the transition to hip fracture, post-hip fracture mortality, and hip fracture-free all-cause death. Population-based cohort from Catalonia, Spain, including all individuals aged 65 to 80 years with a recorded diagnosis of T2DM on 1 January 2006; and non-T2DM matched (up to 2:1) by year of birth, gender, and primary care practice. A total of 44,802 T2DM and 81,233 matched controls (53% women, mean age 72 years old) were followed for a median of 8 years: 23,818 died without fracturing and 3317 broke a hip, of whom 838 subsequently died. Adjusted HRs for hip fracture-free mortality were 1.32 (95% CI 1.28 to 1.37) for men and 1.72 (95% CI 1.65 to 1.79) for women. HRs for hip fracture were 1.24 (95% CI 1.08 to 1.43) and 1.48 (95% CI 1.36 to 1.60), whilst HRs for post-hip fracture mortality were 1.28 (95% CI 1.02 to 1.60) and 1.57 (95% CI 1.31 to 1.88) in men and women, respectively. T2DM individuals are at increased risk of hip fracture, post-hip fracture mortality, and hip fracture-free death. After adjustment, T2DM men were at a 28% higher risk of dying after suffering a hip fracture and women had 57% excess risk of post-hip fracture mortality.


CPF-associated phosphatase activity opposes condensin-mediated chromosome condensation.
Functional links connecting gene transcription and condensin-mediated chromosome condensation have been established in species ranging from prokaryotes to vertebrates. However, the exact nature of these links remains misunderstood. Here we show in fission yeast that the 3' end RNA processing factor Swd2.2, a component of the Cleavage and Polyadenylation Factor (CPF), is a negative regulator of condensin-mediated chromosome condensation. Lack of Swd2.2 does not affect the assembly of the CPF but reduces its association with chromatin. This causes only limited, context-dependent effects on gene expression and transcription termination. However, CPF-associated Swd2.2 is required for the association of Protein Phosphatase 1 PP1(Dis2) with chromatin, through an interaction with Ppn1, a protein that we identify as the fission yeast homologue of vertebrate PNUTS. We demonstrate that Swd2.2, Ppn1 and PP1Dis2 form an independent module within the CPF, which provides an essential function in the absence of the CPF-associated Ssu72 phosphatase. We show that Ppn1 and Ssu72, like Swd2.2, are also negative regulators of condensin-mediated chromosome condensation. We conclude that Swd2.2 opposes condensin-mediated chromosome condensation by facilitating the function of the two CPF-associated phosphatases PP1 and Ssu72.