The role of early childhood education programmes in the promotion of child and adolescent mental health in low- and middle-income countries.
There is growing evidence that early childhood education (ECE) interventions can reduce the loss of developmental potential of disadvantaged children in low- and middle-income countries (LAMIC). Less attention has been paid to the potential of these programmes to prevent child mental health problems and promote child well-being. Peer-reviewed journal articles describing controlled evaluations of ECE interventions in LAMIC were reviewed to identify studies with child mental health outcomes. Studies with proximal outcomes for child mental health including caregiver practices and caregiver mental health were also reviewed. Of 63 studies identified, 21 (33.33%) included child mental health outcomes; 12 of 16 studies with short-term measures showed benefits; 6 studies included a longer-term follow-up and all found benefits; 25 studies included caregiver outcomes: consistent benefits were found for caregiver practices (21 studies) and 6 of 9 studies that measured caregiver mental health reported benefits. Gains to child mental health may be most likely when ECE interventions include three main elements: (i) activities to increase child skills including cognition, language, self-regulation and social-emotional competence; (ii) training caregivers in the skills required to provide a cognitively stimulating and emotionally supportive environment; and (iii) attention to the caregivers' mental health, motivation and self-efficacy. Recommendations for the design and implementation of programmes are provided. ECE interventions are an important component of mental health prevention and promotion in LAMIC, and promoting child and caregiver well-being is a fundamental aspect of interventions to improve child development.


The treatment of intertrochanteric fractures: results using an intramedullary nail with integrated cephalocervical screws and linear compression.
A new device for the treatment of intertrochanteric fractures that uses 2 cephalocervical screws in an integrated mechanism allowing linear intraoperative compression and rotational stability of the head/neck fragment has been developed. The aim of this study was to describe the results using this device for the treatment of stable and unstable intertrochanteric fractures. Prospective, consecutive. Academic Trauma Center. Between March 1, 2005, and July 31, 2006, 100 consecutive patients with an intertrochanteric fracture were treated with a new trochanteric antegrade nail (InterTan; Smith-Nephew, Memphis, TN). All living patients were followed up for a minimum of 1 year postoperatively (range 12-27 months). Clinical and radiographic examinations were performed until healing and at the 1-year anniversary of the index procedure. Healing, pain with ambulation, return to activities of daily living, the modified Harris hip score, and Barthel Index were used to evaluate outcomes. The mean age of the patients was 81.2 (+/-11.3) years. Thirty-seven patients died, 12 were too infirmed for follow-up, and 3 could not be located, leaving 48 patients available for final evaluation. The average surgical time was 41 minutes (13-95 minutes). This rose significantly with the complexity of the fracture (OTA/AO classification: A1 versus A3, P = 0.016). All fractures healed within 16 weeks (range 10-16 weeks). Radiographic analysis at healing revealed no loss of reduction, no uncontrolled collapse of the neck, no nonunions, no femoral shaft fractures, and no implant failures. Two cases in the series were poorly reduced and settled into varus malalignment. There was no varus malposition seen in the remaining 46 fractures. The mean prefracture Harris hip score (75.1 +/- 13.4) was significantly reduced at the time of follow-up (70.3 +/- 14.5, P = 0.003); 58% of the patients recovered their prefracture status. No significant difference was seen for the Barthel Index. The InterTan device appears to be a reliable implant for the treatment of intertrochanteric femoral fractures. Its design provides for stability against rotation and minimizes neck malunions (shortening) through linear intraoperative compression of the head/neck segment to the shaft. As a result of the negligible complication rate and improved clinical outcomes, this implant is now the standard treatment for all intertrochanteric fractures at our institution.


GPs at the Edge: a quantitative description of Irish Rural General Practice.
Currently, more than 1.6 million Irish people live rurally. Rural populations in Ireland are older and have more health needs compared with younger urban areas. Meanwhile, since 1982, the proportion of general practices in rural areas has decreased by 10%. In this study, we look at new survey data to investigate the needs and challenges of rural general practice in Ireland. This study will make use of survey responses from the 2021 membership survey by the Irish College of General Practitioners (ICGP). The anonymous, online, survey was sent by email to the ICGP membership in late 2021, with a series of questions pertaining to practice location, and prior experience of living and working in a rural area designed specifically for this project. A series of statistical tests will be undertaken as appropriate for the data. This study is ongoing; we aim to present data on the demographics of those working in rural general practice and related factors. Previous research has shown that people who grew up or trained in rural areas are more likely to work there after qualifying. As the analysis of this survey continues, it will be important to see if this pattern is evident here as well.


Quantifying indirect evidence in network meta-analysis.
Network meta-analysis enables comprehensive synthesis of evidence concerning multiple treatments and their simultaneous comparisons based on both direct and indirect evidence. A fundamental pre-requisite of network meta-analysis is the consistency of evidence that is obtained from different sources, particularly whether direct and indirect evidence are in accordance with each other or not, and how they may influence the overall estimates. We have developed an efficient method to quantify indirect evidence, as well as a testing procedure to evaluate their inconsistency using Lindsay's composite likelihood method. We also show that this estimator has complete information for the indirect evidence. Using this method, we can assess the degree of consistency between direct and indirect evidence and their contribution rates to the overall estimate. Sensitivity analyses can be also conducted with this method to assess the influences of potentially inconsistent treatment contrasts on the overall results. These methods can provide useful information for overall comparative results that might be biased from specific inconsistent treatment contrasts. We also provide some fundamental requirements for valid inference on these methods concerning consistency restrictions on multi-arm trials. In addition, the efficiency of the developed method is demonstrated based on simulation studies. Applications to a network meta-analysis of 12 new-generation antidepressants are presented. Copyright © 2016 John Wiley & Sons, Ltd.


Area of cartilage accessible to curettage for subsequent arthrodesis of the equine proximal interphalangeal joint. Comparison of conventional and collateral ligament sparing approaches.
This study compared the areas of cartilage accessible for curettage in arthrodesis of the equine proximal interphalangeal joint using the conventional and collateral ligament sparing approaches. For this purpose, forelimbs and hindlimbs of six equine cadavers without radiographic evidence of proximal interphalangeal joint disease were used. One limb of each pair of limbs was randomly assigned to a dissection using a standard approach to the proximal interphalangeal joint including transection of the collateral ligaments, while each contralateral limb was exposed using the same approach but leaving the collateral ligaments intact. Hohmann retractors and Spratt curettes were then used to remove as much articular cartilage as possible. Finally, proximal interphalangeal joints were photographed and image analysis was performed. Using the collateral ligament sparing procedure, the mean percentage of articular cartilage surface removed (41.2%) was significantly less than using the conventional procedure (79.6%) (p <0.01). The difference between forelimbs and hindlimbs was not significant.


Differential transcriptional expression profiles of juvenile and adult calvarial bone.
It has widely been observed that young children are capable of reossifying large calvarial defects, while adults lack this endogenous tissue-engineering capacity. The ability of juvenile animals to regenerate calvarial defects has been investigated in multiple animal models, including mice. In this study, the authors used cDNA microarrays to investigate the expression of osteogenesis-associated genes upstream and downstream of Runx2 in juvenile and adult mouse calvaria. Nonsuture-associated parietal bone discs were harvested from 6-day-old (n = 50) and 60-day-old (n = 35) male CD-1 mice. After separation of the underlying dura mater and overlying pericranium, the calvarial discs were snap-frozen and RNA was extracted from pooled samples of calvaria for microarray analysis. Genes analyzed included cytokines, receptors, and cell-surface and matrix proteins both upstream and downstream of Runx2. Genes associated with the Runx2 pathway had notably higher levels in the juvenile versus adult calvaria. All genes except for osteocalcin were expressed at least twofold higher in the juvenile calvaria. This pattern was validated with quantitative real-time polymerase chain reaction. In addition, mRNA for potent osteoinductive growth factors was present at higher levels in the juvenile compared with the adult calvaria. These findings reflect a genomic environment of active osteoblast differentiation and ossification in the juvenile calvaria compared with the adult "quiescent" calvarial tissue. These data suggest that a decreased osteogenic potential of adult calvarial osteoblasts may, in part, explain the inability of adult animals to heal calvarial defects.


Genotypic and phenotypic analyses reveal distinct population structures and ecotypes for sugar beet-associated Pseudomonas in Oxford and Auckland.
Fluorescent pseudomonads represent one of the largest groups of bacteria inhabiting the surfaces of plants, but their genetic composition in planta is poorly understood. Here, we examined the population structure and diversity of fluorescent pseudomonads isolated from sugar beet grown at two geographic locations (Oxford, United Kingdom and Auckland, New Zealand). To seek evidence for niche adaptation, bacteria were sampled from three types of leaves (immature, mature, and senescent) and then characterized using a combination of genotypic and phenotypic analysis. We first performed multilocus sequence analysis (MLSA) of three housekeeping genes (gapA, gltA, and acnB) in a total of 152 isolates (96 from Oxford, 56 from Auckland). The concatenated sequences were grouped into 81 sequence types and 22 distinct operational taxonomic units (OTUs). Significant levels of recombination were detected, particularly for the Oxford isolates (rate of recombination to mutation (r/m) = 5.23 for the whole population). Subsequent ancestral analysis performed in STRUCTURE found evidence of six ancestral populations, and their distributions significantly differed between Oxford and Auckland. Next, their ability to grow on 95 carbon sources was assessed using the Biolog™ GN2 microtiter plates. A distance matrix was generated from the raw growth data (A 660) and subjected to multidimensional scaling (MDS) analysis. There was a significant correlation between substrate utilization profiles and MLSA genotypes. Both phenotypic and genotypic analyses indicated presence of a geographic structure for strains from Oxford and Auckland. Significant differences were also detected for MLSA genotypes between strains isolated from immature versus mature/senescent leaves. The fluorescent pseudomonads thus showed an ecotypic population structure, suggestive of adaptation to both geographic conditions and local plant niches.


Diamonds and the african lithosphere.
Data and inferences drawn from studies of diamond inclusions, xenocrysts, and xenoliths in the kimberlites of southern Africa are combined to characterize the structure of that portion of the Kaapvaal craton that lies within the mantle. The craton has a root composed in large part of peridotites that are strongly depleted in basaltic components. The asthenosphere boundary shelves from depths of 170 to 190 kilometers beneath the craton to approximately 140 kilometers beneath the mobile belts bordering the craton on the south and west. The root formed earlier than 3 billion years ago, and at that time ambient temperatures in it were 900 degrees to 1200 degrees C; these temperatures are near those estimated from data for xenoliths erupted in the Late Cretaceous or from present-day heat-flow measurements. Many of the diamonds in southern Africa are believed to have crystallized in this root in Archean time and were xenocrysts in the kimberlites that brought them to the surface.


Ephedrine versus phenylephrine as a vasopressor for spinal anaesthesia-induced hypotension in parturients undergoing high-risk caesarean section: meta-analysis, meta-regression and trial sequential analysis.
Phenylephrine is the preferred vasopressor for the prevention and treatment of spinal anaesthesia-induced hypotension during caesarean section, because studies on low-risk elective patients found it to have a less detrimental effect on umbilical artery pH compared with ephedrine. However, limited data exist from high-risk parturients and parturients with uteroplacental insufficiency. We systematically searched for randomised, controlled, double-blinded trials of these two vasopressors in high-risk caesarean sections. We applied conventional meta-analysis, trial sequential analysis, computing the required information size that would exclude type I and II errors, contour-enhanced funnel plot testing for publication bias, meta-regression to assess the dose-response relationship, and the Grading of Recommendations Assessment, Development, and Evaluation system (GRADE). The incidence of fetal acidosis (umbilical arterial pH <7.2) was the primary outcome. Eight trials (712 patients) with low risk of bias were identified. Pooling six studies of patients with preeclampsia and other reasons for fetal compromise, as well as subgroup analysis of the preeclampsia studies, revealed no significant differences in the incidence of fetal acidosis. Trial sequential analysis showed that the required information size was not reached. The funnel plot was not suggestive of publication bias. Meta-regression showed no dose-response relationship. The GRADE score was moderate quality. Despite several studies and a large number of patients there was insufficient evidence to make a recommendation for choice of vasopressor in high-risk caesarean section. Trials with adequate power to detect differences in the incidence of fetal acidosis between ephedrine and phenylephrine are required to provide evidence-based guidance.


A Pragmatic Application of the RE-AIM Framework for Evaluating the Implementation of Physical Activity as a Standard of Care in Health Systems.
Exercise is Medicine (EIM) is an initiative that seeks to integrate physical activity assessment, prescription, and patient referral as a standard in patient care. Methods to assess this integration have lagged behind its implementation. The purpose of this work is to provide a pragmatic framework to guide health care systems in assessing the implementation and impact of EIM. A working group of experts from health care, public health, and implementation science convened to develop an evaluation model based on the RE-AIM (Reach, Effectiveness, Adoption, Implementation, and Maintenance) framework. The working group aimed to provide pragmatic guidance on operationalizing EIM across the different RE-AIM dimensions based on data typically available in health care settings. The Reach of EIM can be determined by the number and proportion of patients that were screened for physical inactivity, received brief counseling and/or a physical activity prescription, and were referred to physical activity resources. Effectiveness can be assessed through self-reported changes in physical activity, cardiometabolic biometric factors, incidence/burden of chronic disease, as well as health care utilization and costs. Adoption includes assessing the number and representativeness of health care settings that adopt any component of EIM, and Implementation involves assessing the extent to which health care teams implement EIM in their clinic. Finally, Maintenance involves assessing the long-term effectiveness (patient level) and sustained implementation (clinic level) of EIM in a given health care setting. The availability of a standardized, pragmatic, evaluation framework is critical in determining the impact of implementing EIM as a standard of care across health care systems.


Street-level noise in an urban setting: assessment and contribution to personal exposure.
The urban soundscape, which represents the totality of noise in the urban setting, is formed from a wide range of sources. One of the most ubiquitous and least studied of these is street-level (i.e., sidewalk) noise. Mainly associated with vehicular traffic, street level noise is hard to ignore and hard to escape. It is also potentially dangerous, as excessive noise from any source is an important risk factor for adverse health effects. This study was conducted to better characterize the urban soundscape and the role of street level noise on overall personal noise exposure in an urban setting. Street-level noise measures were obtained at 99 street sites located throughout New York City (NYC), along with data on time, location, and sources of environmental noise. The relationship between street-level noise measures and potential predictors of noise was analyzed using linear and logistic regression models, and geospatial modeling was used to evaluate spatial trends in noise. Daily durations of street-level activities (time spent standing, sitting, walking and running on streets) were estimated via survey from a sample of NYC community members recruited at NYC street fairs. Street-level noise measurements were then combined with daily exposure durations for each member of the sample to estimate exposure to street noise, as well as exposure to other sources of noise. The mean street noise level was 73.4 dBA, with substantial spatial variation (range 55.8-95.0 dBA). Density of vehicular (road) traffic was significantly associated with excessive street level noise levels. Exposure duration data for street-level noise and other common sources of noise were collected from 1894 NYC community members. Based on individual street-level exposure estimates, and in consideration of all other sources of noise exposure in an urban population, we estimated that street noise exposure contributes approximately 4% to an average individual's annual noise dose. Street-level noise exposure is a potentially important source of overall noise exposure, and the reduction of environmental sources of excessive street- level noise should be a priority for public health and urban planning.


Dimensionality and Reliability of the Central Sensitization Inventory in a Pooled Multicountry Sample.
Central sensitization (CS) involves the amplification of neural signaling within the central nervous system, which evokes pain hypersensitivity. The Central Sensitization Inventory (CSI) assesses 25 overlapping health-related symptom dimensions that have been reported to be associated with CS-related disorders. Previous studies have reported satisfactory test-retest reliability and internal consistency, but factor analyses have exhibited conflicting results in different language versions. The purpose of this cross-sectional study was to thoroughly examine the dimensionality and reliability of the CSI, with pooled data from 1,987 individuals, collected in several countries. The principal component analysis suggested that 1 general factor of CS best described the structure. A subsequent confirmatory factor analysis revealed that a bifactor model, which accounted for the covariance among CSI items, with regard to 1 general factor and 4 orthogonal factors, fit the CSI structure better than the unidimensional and the 4-factor models. Additional analyses indicated substantial reliability for the general factor (ie, Cronbach α = .92; ω = .95; and ω hierarchical = .89). Reliability results for the 4 specific factors were considered too low to be used for subscales. The results of this study clearly suggest that only total CSI scores should be used and reported. As far as we know, this is the first study that has examined the factor structure and reliability of the CSI in a large multicountry sample. The CSI is currently considered the leading self-report measure of CS-related symptoms worldwide.


Bioimpedance spectroscopy for the estimation of body fluid volumes in mice.
Conventional indicator dilution techniques for measuring body fluid volume are laborious, expensive, and highly invasive. Bioimpedance spectroscopy (BIS) may be a useful alternative due to being rapid, minimally invasive, and allowing repeated measurements. BIS has not been reported in mice; hence we examined how well BIS estimates body fluid volume in mice. Using C57/Bl6 mice, the BIS system demonstrated <5% intermouse variation in total body water (TBW) and extracellular (ECFV) and intracellular fluid volume (ICFV) between animals of similar body weight. TBW, ECFV, and ICFV differed between heavier male and lighter female mice; however, the ratio of TBW, ECFV, and ICFV to body weight did not differ between mice and corresponded closely to values in the literature. Furthermore, repeat measurements over 1 wk demonstrated <5% intramouse variation. Default resistance coefficients used by the BIS system, defined for rats, produced body composition values for TBW that exceeded body weight in mice. Therefore, body composition was measured in mice using a range of resistance coefficients. Resistance values at 10% of those defined for rats provided TBW, ECFV, and ICFV ratios to body weight that were similar to those obtained by conventional isotope dilution. Further evaluation of the sensitivity of the BIS system was determined by its ability to detect volume changes after saline infusion; saline provided the predicted changes in compartmental fluid volumes. In summary, BIS is a noninvasive and accurate method for the estimation of body composition in mice. The ability to perform serial measurements will be a useful tool for future studies.


Exploring and building autism service capacity in rural and remote regions: Participatory action research in rural Alberta and British Columbia, Canada.
Based in participatory action research, this project had the aim of building capacity in treatment and support for individuals and families impacted by autism spectrum disorder in remote and rural communities of Canada. Communities were selected based on their rurality and willingness to engage in change efforts for enhanced service delivery within their region. Fifteen discussion groups with key stakeholders were convened in seven communities with ~200 community stakeholders. Based on analyses of these data from the stakeholders, themes were distilled through interpretive description, which in turn were presented to community stakeholders for reflection and collective action. Findings indicate broad thematic domains consisting of: insufficient services, protective factors in community, change efforts via collectivity within community, limitations and benefits of residing in rural communities relative to care associated with autism spectrum disorder, a sense of "community" in rural contexts, and engaging in focused dialogue as a pathway to advancement. Opportunities for building capacity for support in autism spectrum disorder emerged within intersecting layers of leadership, contextual factors, and community collaboration. Consistent with participatory action research principles, emerging local knowledge was supported with strategies for improved autism spectrum disorder service development.


Implementation of an antibiotic nomogram improves postoperative antibiotic utilization and safety in patients undergoing coronary artery bypass grafting.
Routine, initial, empiric vancomycin dosing by clinicians in postoperative coronary artery bypass grafting (CABG) patients was identified as a potential patient safety issue in the Cardiovascular Intensive Care Unit (CVICU) because the rate of postoperative acute renal insufficiency (ARI) and average patient Body Mass Index (BMI) > 35 kg/m2 were significantly higher in our institution than those of the Society of Thoracic Surgeons (STS) database. A vancomycin dosing nomogram was derived from the local patient population in the attempt to improve patient safety by convincing clinicians to use an evidence-based approach to vancomycin prescription. We analyzed two different treatment strategies that were applied consecutively to an intensive care unit population. CABG patients dosed empirically with vancomycin (group 1, pre-nomogram) were compared with CABG patients dosed using a vancomycin dosing nomogram (group 2, post-nomogram) derived from the hospital population using an Internet program that facilitated creation of a local nomogram. The two groups were analyzed as to age, sex, body mass index, creatinine clearance, and vancomycin dosage using logistic regression and testing for continuous and categorical variables. Nomogram use decreased the number of patients receiving the customary dose of one gram every 12 hours in those group 2 patients with diminished CrCl as compared with those in group 1 with diminished CrCl (group 2, 2/21 vs. group 1, 14/21, p < .0001), as well as in those with a normal creatinine clearance, (group 2, 2/15 vs. group 1, 26/34, p < .0001). Therefore, nomogram use affected the customary dose of one g vancomycin every 12 hours between the two groups overall (group 1, 40/55 vs. group 2, 4/36, p < .001), whereby 32/36 (88.9%) of group 2 patients had their dosing altered when compared to what would have been formerly prescribed, p < .0001. Furthermore, nomogram use resulted in fewer doses of antibiotics per year resulting in a cost savings to the hospital with no increase in the rates of infection. Implementation of the nomogram resulted in a more appropriate antibiotic utilization, regardless of creatinine clearance, that decreased costs without increasing infection rates.


Disinfection of Needleless Connector Hubs: Clinical Evidence Systematic Review.
Background. Needleless connectors (NC) are used on virtually all intravascular devices, providing an easy access point for infusion connection. Colonization of NC is considered the cause of 50% of postinsertion catheter-related infections. Breaks in aseptic technique, from failure to disinfect, result in contamination and subsequent biofilm formation within NC and catheters increasing the potential for infection of central and peripheral catheters. Methods. This systematic review evaluated 140 studies and 34 abstracts on NC disinfection practices, the impact of hub contamination on infection, and measures of education and compliance. Results. The greatest risk for contamination of the catheter after insertion is the NC with 33-45% contaminated, and compliance with disinfection as low as 10%. The optimal technique or disinfection time has not been identified, although scrubbing with 70% alcohol for 5-60 seconds is recommended. Studies have reported statistically significant results in infection reduction when passive alcohol disinfection caps are used (48-86% reduction). Clinical Implications. It is critical for healthcare facilities and clinicians to take responsibility for compliance with basic principles of asepsis compliance, to involve frontline staff in strategies, to facilitate education that promotes understanding of the consequences of failure, and to comply with the standard of care for hub disinfection.


A population based study of seasonality of skin and soft tissue infections: implications for the spread of CA-MRSA.
Methicillin resistant Staphylococcus aureus (MRSA) is currently a major cause of skin and soft tissue infections (SSTI) in the United States. Seasonal variation of MRSA infections in hospital settings has been widely observed. However, systematic time-series analysis of incidence data is desirable to understand the seasonality of community acquired (CA)-MRSA infections at the population level. In this paper, using data on monthly SSTI incidence in children aged 0-19 years and enrolled in Medicaid in Maricopa County, Arizona, from January 2005 to December 2008, we carried out time-series and nonlinear regression analysis to determine the periodicity, trend, and peak timing in SSTI incidence in children at different age: 0-4 years, 5-9 years, 10-14 years, and 15-19 years. We also assessed the temporal correlation between SSTI incidence and meteorological variables including average temperature and humidity. Our analysis revealed a strong annual seasonal pattern of SSTI incidence with peak occurring in early September. This pattern was consistent across age groups. Moreover, SSTIs followed a significantly increasing trend over the 4-year study period with annual incidence increasing from 3.36% to 5.55% in our pediatric population of approximately 290,000. We also found a significant correlation between the temporal variation in SSTI incidence and mean temperature and specific humidity. Our findings could have potential implications on prevention and control efforts against CA-MRSA.


The Moderating Influence of International Courts on Social Movements: Evidence from the IVF Case Against Costa Rica.
Feminists and religious conservatives across the globe have increasingly turned to courts in their battles over abortion. Yet while a significant literature analyzes legal mobilization on abortion issues, it tends to focus predominantly on domestic scenarios. In this article, we consider the effects of this contentious engagement of pro-choice and anti-abortion movements in international human rights fora, asking what happens to social movement claims when they reach international human rights courts. We answer the question through a detailed description of a single case, Gretel Artavia Murillo et al. v. Costa Rica, decided by the Inter-American Court of Human Rights in 2012 but with ongoing repercussions for abortion rights, given its authoritative interpretation of embryonic right to life. Through our analysis of Artavia Murillo, we show how legal mobilization before international human rights courts moderates social movement claims within the legal arena, as rivals respond to one another and argue within the frame of courts' norms and language.


Assessment of metabolic diversity within the intestinal microbiota from healthy humans using combined molecular and cultural approaches.
The human gut harbours a wide range of bacterial communities that play key roles in supplying nutrients and energy to the host through anaerobic fermentation of dietary components and host secretions. This fermentative process involves different functional groups of microorganisms linked in a trophic chain. Although the diversity of the intestinal microbiota has been studied extensively using molecular techniques, the functional aspects of this biodiversity remain mostly unexplored. The aim of the present work was to enumerate the principal metabolic groups of microorganisms involved in the fermentative process in the gut of healthy humans. These functional groups of microorganisms were quantified by a cultural approach, while the taxonomic composition of the microbiota was assessed by in situ hybridization on the same faecal samples. The functional groups of microorganisms that predominated in the gut were the polysaccharide-degrading populations involved in the breakdown of the most readily available exogenous and endogenous substrates and the predominant butyrate-producing species. Most of the functional groups of microorganisms studied appeared to be present at rather similar levels in all healthy volunteers, suggesting that optimal numbers of these various bacterial groups are crucial for efficient gut fermentation, as well as for host nutrition and health. Significant interindividual differences were, however, confirmed with respect to the numbers of methanogenic archaea, filter paper-degrading and acetogenic bacteria and the products formed by lactate-utilizing bacteria.


The implementation of a kidney exchange program does not induce a need for additional psychosocial support.
The Dutch kidney exchange donation program started in January 2004. A literature review has shown that several factors of the exchange program could influence the psychological well being of participants, such as the loss of the possibility of a 'medical excuse' for unwilling donors and the issue of anonymity. However, these factors have not been the subject of empirical study yet. We therefore studied these factors to determine whether additional psychosocial support is necessary for donors and recipients in the Dutch kidney exchange program. We used structured interviews for all 48 donors and recipients that had undergone exchange donation/transplantation in 2004. A psychologist interviewed the participants before and 3 months after transplantation. We included a comparison group of 48 donors and recipients participating in the regular living kidney donation program. Donors did not experience additional pressure to donate due to the exchange donation. Most participants (69%) preferred anonymity between the couples. Ten percentage needed additional emotional support. In this respect the exchange group did not differ from the comparison group. We conclude that the psychosocial support offered to exchange couples can be comparable with the support normally offered to participants in the regular living kidney donation program.


Systemic lipopolysaccharide protects the brain from ischemic injury by reprogramming the response of the brain to stroke: a critical role for IRF3.
Lipopolysaccharide (LPS) preconditioning provides neuroprotection against subsequent cerebral ischemic injury through activation of its receptor, Toll-like receptor 4 (TLR4). Paradoxically, TLR activation by endogenous ligands after ischemia worsens stroke damage. Here, we define a novel, protective role for TLRs after ischemia in the context of LPS preconditioning. Microarray analysis of brains collected 24 h after stroke revealed a unique set of upregulated genes in LPS-pretreated animals. Promoter analysis of the unique gene set identified an overrepresentation of type I interferon (IFN)-associated transcriptional regulatory elements. This finding suggested the presence of type I IFNs or interferon regulatory factors (IRFs), which upregulate interferon-stimulated genes. Upregulation of IFNbeta was confirmed by real-time reverse transcription-PCR. Direct administration of IFNbeta intracerebroventricularly at the time of stroke was sufficient for neuroprotection. TLR4 can induce both IFNbeta and interferon-stimulated genes through its adapter molecule Toll/interleukin receptor domain-containing adaptor-inducing IFNbeta (TRIF) and the IRF3 transcription factor. We show in oxygen glucose deprivation of cortical neurons, an in vitro model of stroke, that activation of TRIF after stroke reduces neuronal death. Furthermore, mice lacking IRF3 were not protected by LPS preconditioning in our in vivo model. Our studies constitute the first demonstration of the neuroprotective capacity of TRIF/IRF3 signaling and suggest that interferon-stimulated genes, whether induced by IFNbeta or by enhanced TLR signaling to IRF3, are a potent means of protecting the brain against ischemic damage.


MR imaging findings in a dog with lumbar ganglion cysts.
Intraspinal cysts of the L6-L7 and L7-S1 articular process joints in a six-year-old neutered female German Shepherd Dog were diagnosed using magnetic resonance (MR) imaging. Histopathology provided a diagnosis of ganglion cysts. Clinical, laboratory, radiographic and MR imaging findings are described. Briefly, radiographic findings revealed lumbarization of the first sacral vertebra, and fusion of the first caudal vertebra to the sacrum. In addition, spondylosis and articular process osteoarthrosis occurred at L6-L7 and L7-S1. MR imaging revealed multiple, well encapsulated structures ranging in size from 3-10 mm in diameter which were found to arise from the L6-L7 and L7-S1 articular process joints. These cysts had signal intensities that varied from hyperintense to the cerebrospinal fluid (CSF) on T1 weighted images to isointense to CSF on T2 weighted images. Decompressive surgery in conjunction with arthrodesis of these joints resulted in resolution of clinical signs. The dog remained pain-free 1 1/2 years following surgical therapy.


Comparison of media for recovery of total coliform bacteria from chemically treated water.
Five broth media and two solid media were compared for their ability to quantitatively recover total coliform bacteria from chemically treated water. M-Endo LES and mT7 media were used in the membrane filter technique. Lauryl tryptose broth, lactose broth, presence-absence broth, lactose broth with twice the amount of lactose, and lauryl tryptose broth with twice the amount of sodium lauryl sulfate were used in the fermentation tube procedure. The differences in recovery were not significant for the five broth media and M-Endo LES agar. The M-Endo LES and mT7 media were not significantly different; however, the five broth media did yield significantly higher counts than mT7.


A novel artificial synapse with dual modes using bilayer graphene as the bottom electrode.
Resistive Random Access Memory (RRAM) shows great potential to be used as an artificial synapse for neuromorphic applications. The resistance can be gradually reduced during reset, which can enable enough states to mimic the "forgetting" process. However, the abrupt set (Mode I) cannot generate enough states to mimic the "learning" process, which results in depression-only behavior. In this work, we introduce another mode (Mode II) in an Al/AlOx/graphene 'RRAM' stack by using oxygen vacancies as trapping centers and bottom electrode bilayer graphene as the channel material. In this way, since the pulse can gradually create the oxygen vacancies, post-synaptic current (PSC) can be gradually potentiated or depressed. By introducing Mode II, we can realize 166 potentiation states, which is higher than the previously reported conventional RRAM with insufficient potentiation states due to the abrupt set. Moreover, Mode II can help realize an inhibitory synapse. By combining modes I and II, we can realize both excitatory and inhibitory synapses in a single device. This work shows great potential to enable neuromorphic computations with greater learning and reconfigurability.


Prescribing sea water quality criteria for arsenic, cadmium and lead through species sensitivity distribution.
Water quality standards are essential for regulation of contaminants in marine environment. Seawater quality criteria (SWQC) for arsenic (As), cadmium (Cd) and lead (Pb) have not been developed for India. The aim of this study is to derive the SWQC for the metals based on Species Sensitivity Distribution (SSD). Eight species of sensitive marine organisms belonging to five phyla were assessed for their sensitivity to toxicity of As, Cd and Pb. Median effective concentrations (EC50) and Median Lethal Concentrations (LC50) were derived from the acute toxicity bio-assays. No Observed Effect Concentrations (NOEC), Lowest Observed Effect Concentrations (LOEC) and chronic values were derived from chronic toxicity bio-assays. Diatoms were more sensitive to As with 96 h EC50 of 0.1 mg/l and copepods were more sensitive to Cd and Pb with 96 h EC50 of 0.019 mg/l and 0.05 mg/l respectively. Estimated NOECs ranged from 4.87 to 21.55 µg/l of As, 1.0 to 120 µg/l of Cd and 5.67 to 91.67 µg/l of Pb. Similarly, chronic values (µg/l) were in the range of 6.71-26.1, 1.38-170, and 7.67-91.67 of As, Cd and Pb respectively. The Criterion Maximum Concentration (CMC), Criterion Continuous Concentration (CCC) and Predicted No Effect Concentration (PNEC) values were prescribed as SWQC. The CMC (µg/l) of 19, 1.7 and 17 for As, Cd, and Pb were derived respectively for acute exposure during accidental marine outfalls. The CCC (µg/l) for As was 4.6, 1.1 for Cd and 5.9 for Pb are recommended as SWQC for protection of 95% of marine organisms. PNEC (µg/l) of 3.8 for As, 0.92 for Cd and 4.3 for Pb are suggested for highly disturbed ecosystems, shell fishing and mariculture uses of water bodies. These values are recommended as a baseline for site specific water quality criteria for the coastal waters of the country.


Gene expression profiling of human whole blood samples with the Illumina WG-DASL assay.
Microarray-based gene expression analysis of peripheral whole blood is a common strategy in the development of clinically relevant biomarker panels for a variety of human diseases. However, the results of such an analysis are often plagued by decreased sensitivity and reliability due to the effects of relatively high levels of globin mRNA in whole blood. Globin reduction assays have been shown to overcome such effects, but they require large amounts of total RNA and may induce distinct gene expression profiles. The Illumina whole genome DASL assay can detect gene expression levels using partially degraded RNA samples and has the potential to detect rare transcripts present in highly heterogeneous whole blood samples without the need for globin reduction. We assessed the utility of the whole genome DASL assay in an analysis of peripheral whole blood gene expression profiles. We find that gene expression detection is significantly increased with the use of whole genome DASL compared to the standard IVT-based direct hybridization. Additionally, globin-probe negative whole genome DASL did not exhibit significant improvements over globin-probe positive whole genome DASL. Globin reduction further increases the detection sensitivity and reliability of both whole genome DASL and IVT-based direct hybridization with little effect on raw intensity correlations. Raw intensity correlations between total RNA and globin reduced RNA were 0.955 for IVT-based direct hybridization and 0.979 for whole genome DASL. Overall, the detection sensitivity of the whole genome DASL assay is higher than the IVT-based direct hybridization assay, with or without globin reduction, and should be considered in conjunction with globin reduction methods for future blood-based gene expression studies.


Syndrome complex of bone marrow failure and pulmonary fibrosis predicts germline defects in telomerase.
Mutations in the essential telomerase components hTERT and hTR cause dyskeratosis congenita, a bone marrow failure syndrome characterized by mucocutaneous features. Some (~ 3%) sporadic aplastic anemia (AA) and idiopathic pulmonary fibrosis cases also carry mutations in hTERT and hTR. Even though it can affect clinical outcome, because the mutation frequency is rare, genetic testing is not standard. We examined whether the cooccurrence of bone marrow failure and pulmonary fibrosis in the same individual or family enriches for the presence of a telomerase mutation. Ten consecutive individuals with a total of 36 family members who fulfilled these criteria carried a germline mutant telomerase gene (100%). The mean age of onset for individuals with AA was significantly younger than that for those with pulmonary fibrosis (14 vs 51; P < .0001). Families displayed autosomal dominant inheritance and there was an evolving pattern of genetic anticipation, with the older generation primarily affected by pulmonary fibrosis and successive generations by bone marrow failure. The cooccurrence of AA and pulmonary fibrosis in a single patient or family is highly predictive for the presence of a germline telomerase defect. This diagnosis affects the choice of bone marrow transplantation preparative regimen and can prevent morbidity.


New records and detailed distribution and abundance of selected arthropod species collected between 1999 and 2011 in Azorean native forests.
In this contribution we present detailed distribution and abundance data for arthropod species identified during the BALA - Biodiversity of Arthropods from the Laurisilva of the Azores (1999-2004) and BALA2 projects (2010-2011) from 18 native forest fragments in seven of the nine Azorean islands (all excluding Graciosa and Corvo islands, which have no native forest left). Of the total 286 species identified, 81% were captured between 1999 and 2000, a period during which only 39% of all the samples were collected. On average, arthropod richness for each island increased by 10% during the time frame of these projects. The classes Arachnida, Chilopoda and Diplopoda represent the most remarkable cases of new island records, with more than 30% of the records being novelties. This study stresses the need to expand the approaches applied in these projects to other habitats in the Azores, and more importantly to other less surveyed taxonomic groups (e.g. Diptera and Hymenoptera). These steps are fundamental for getting a more accurate assessment of biodiversity in the archipelago.


Chlorpromazine alters cochlear mechanics and amplification: in vivo evidence for a role of stiffness modulation in the organ of corti.
Although prestin-mediated outer hair cell (OHC) electromotility provides mechanical force for sound amplification in the mammalian cochlea, proper OHC stiffness is required to maintain normal electromotility and to transmit mechanical force to the basilar membrane (BM). To investigate the in vivo role of OHC stiffness in cochlear amplification, chlorpromazine (CPZ), an antipsychotic drug that alters OHC lateral wall biophysics, was infused into the cochleae in living guinea pigs. The effects of CPZ on cochlear amplification and OHC electromotility were observed by measuring the acoustically and electrically evoked BM motions. CPZ significantly reduced cochlear amplification as measured by a decline of the acoustically evoked BM motion near the best frequency (BF) accompanied by a loss of nonlinearity and broadened tuning. It also substantially reduced electrically evoked BM vibration near the BF and at frequencies above BF (< or =80 kHz). The high-frequency notch (near 50 kHz) in the electrically evoked BM response shifted toward higher frequency in a CPZ concentration-dependent manner with a corresponding phase change. In contrast, salicylate resulted in a shift in this notch toward lower frequency. These results indicate that CPZ reduces OHC-mediated cochlear amplification probably via its effects on the mechanics of the OHC plasma membrane rather than via a direct effect on the OHC motor, prestin. Through modeling, we propose that with a combined OHC somatic and hair bundle forcing, the upward-shift of the approximately 50-kHz notch in the electrically-evoked BM motion may indicate stiffness increase of the OHCs that is responsible for the reduced cochlear amplification.


Statin use, intensity, and 3-year clinical outcomes among older patients with coronary artery disease.
Clinical trial evidence suggests that statin therapy reduces adverse clinical events and provides even greater benefit at high-intensity doses in coronary artery disease (CAD) patients, yet few studies have examined this in clinical practice. We linked detailed in-hospital data (2005-2009) on 15,729 Get With The Guidelines-CAD patients ≥65 years prescribed statins to Centers for Medicare and Medicaid Services claims. High-intensity statin therapy was defined as discharge prescription of atorvastatin ≥40 mg, rosuvastatin ≥20 mg, or simvastatin 80 mg. We used Kaplan-Meier curves to calculate all-cause mortality, major adverse cardiovascular events (MACEs), and all-cause readmission at 3 years postdischarge; log-rank tests to compare survival via overall statin use and intensity; and Cox proportional hazards regression with inverse propensity weighting to evaluate adjusted rates of adverse events over 3 years postdischarge. Of 35,903 patients meeting inclusion criteria, 24,367 (67.9%) were discharged on statin. Of 15,729 patients with statin intensity information, 4488 (28.5%) received high-intensity therapy; these recipients were more often younger, male, and had acute myocardial infarction. After inverse propensity weighting adjustment, statin use was associated with significantly lower hazards of mortality (hazard ratio 0.89, 95% CI 0.84-0.93) and MACE (0.92, 0.88-0.96), but not readmission (1.01, 0.97-1.04). High-intensity (vs low/moderate) use was not associated with lower risk of all-cause mortality (1.07, 1.00-1.14), MACE (1.05, 0.99-1.11), or readmission (1.05, 1.00-1.10). Clinically relevant subgroups had similar results. In older hospitalized CAD patients, use of statin therapy at discharge was associated with improved long-term outcomes. Consistent with current American College of Cardiology/American Heart Association cholesterol guideline recommendations supporting moderate- rather than high-intensity statin therapy in CAD patients >75 years, high-intensity statin therapy was not associated with incremental benefit in this older population.


From organoids to organs: Bioengineering liver grafts from hepatic stem cells and matrix.
Due to the complex function and structure of the liver, resourceful solutions for treating end-stage liver disease are required. Currently, liver transplantation is the only curative therapeutic option. However, due to a worldwide donor shortage, researchers have been looking in other fields for alternative sources of transplantable liver tissue. Recent advances in our understanding of liver physiology, stem cell and matrix biology, have accelerated tissue engineering research. Most notable is the discovery of a culture system to grow liver-like organoids from human hepatic stem cells. The extensive expansion capacity of these stem cells has contributed greatly to the availability of hepatocyte-like cells for tissue engineering. In addition, new techniques are explored to obtain biological liver scaffolds from full size donor organs. This review summarizes these state-of-art techniques which may lay the groundwork towards re-creating transplantable tissue from autologous or allogenic stem cells in the coming decade.


Sex differences in risk factors for myocardial infarction: cohort study of UK Biobank participants.
To investigate sex differences in risk factors for incident myocardial infarction (MI) and whether they vary with age. Prospective population based study. UK Biobank. 471 998 participants (56% women; mean age 56.2) with no history of cardiovascular disease. Incident (fatal and non-fatal) MI. 5081 participants (1463 (28.8%) of whom were women) had MI over seven years' mean follow-up, resulting in an incidence per 10 000 person years of 7.76 (95% confidence interval 7.37 to 8.16) for women and 24.35 (23.57 to 25.16) for men. Higher blood pressure indices, smoking intensity, body mass index, and the presence of diabetes were associated with an increased risk of MI in men and women, but associations were attenuated with age. In women, systolic blood pressure and hypertension, smoking status and intensity, and diabetes were associated with higher hazard ratios for MI compared with men: ratio of hazard ratios 1.09 (95% confidence interval 1.02 to 1.16) for systolic blood pressure, 1.55 (1.32 to 1.83) for current smoking, 2.91 (1.56 to 5.45) for type 1 diabetes, and 1.47 (1.16 to 1.87) for type 2 diabetes. There was no evidence that any of these ratios of hazard ratios decreased with age (P>0.2). With the exception of type 1 diabetes, the incidence of MI was higher in men than in women for all risk factors. Although the incidence of MI was higher in men than in women, several risk factors were more strongly associated with MI in women compared with men. Sex specific associations between risk factors and MI declined with age, but, where it occurred, the higher relative risk in women remained. As the population ages and the prevalence of lifestyle associated risk factors increase, the incidence of MI in women will likely become more similar to that in men.


Pachyonychia congenita in pediatric patients: natural history, features, and impact.
IMPORTANCE Nail dystrophy in early childhood often suggests a diagnosis of pachyonychia congenita (PC). No previous investigation has focused on the early signs of PC and the natural course of the disease. OBJECTIVES To determine the course of pediatric PC, correlate the disease course with the clinical appearance and specific gene mutations, and assess the effect of pediatric PC on quality of life. DESIGN, SETTING, AND PARTICIPANTS One hundred one patients or families with genetically confirmed PC from the International Pachyonychia Congenita Research Registry who completed a survey on the general clinical features of PC and an auxiliary questionnaire on the clinical presentation and quality-of-life issues related to pediatric PC. EXPOSURE Individuals with pachyonychia congenita. MAIN OUTCOMES AND MEASURES Completion of both surveys. RESULTS At birth, toenail changes were present in 47.5% of patients; fingernail changes in 40.6%; and plantar keratoderma in 6.9%. By 5 years of age, these 3 key manifestations were found in 81.2%, 74.2%, and 75.3%, respectively, of individuals with genotype-confirmed PC. The correct diagnosis was made during the first year of life in 26.7% of patients despite the presence of toenail dystrophy in more than 65.3%. Clinical differences that distinguished PC subtypes included (1) later onset and less frequent occurrence of nail dystrophy and keratoderma in PC-K6b, PC-K6c, and PC-K16; (2) concurrent fingernail and toenail thickening in PC-K6a and PC-K17; (3) more palmar keratoderma in PC-K16; (4) cysts primarily in PC-K17 and follicular hyperkeratoses primarily in PC-K6a; (5) hoarseness and/or oral leukokeratoses in the first year of life most often in PC-K6a; and (6) natal teeth exclusively in PC-K17. Among pediatric patients, PC affected the social interactions and function of adolescents most profoundly. CONCLUSIONS AND RELEVANCE Among patients with a detectable mutation, PC manifests with nail thickening and plantar keratoderma before school age in more than three-quarters of affected children, allowing early diagnosis. The highly visible nail changes and painful plantar thickening exert a psychosocial effect on most affected adolescents. Phenotype-genotype correlations in children with PC validate the new classification based on the affected gene.


Intensifying therapy for hypertension despite suboptimal adherence.
More intensive management can improve control blood pressure (BP) in hypertensive patients. However, many would posit that treatment intensification (TI) is not beneficial in the face of suboptimal adherence. We investigated whether the effect of TI on BP varies by adherence. We enrolled 819 patients with hypertension, managed in primary care at an academically-affiliated inner-city hospital. We used the following formula to characterize TI: (visits with a medication change-visits with elevated BP)/total visits. Adherence was characterized using electronic monitoring devices ("MEMS caps"). Patients who returned their MEMS caps (671) were divided into quartiles of adherence, whereas patients who did not return their MEMS caps (148) had "missing" adherence. We examined the relationship between TI and the final systolic blood pressure (SBP), controlling for patient-level covariates. In the entire sample, each additional therapy increase per 10 visits predicted a 2.0 mm Hg decrease in final SBP (P<0.001). After stratifying by adherence, in the "best" adherence quartile each therapy increase predicted a 2.1-mm Hg decrease in final SBP, followed by 1.8 for the "next-best" adherence quartile, 2.3 in the third quartile, and 2.4 in the "worst" adherence quartile. The effect size for patients with "missing" adherence was 1.6 mm Hg. The differences between the group with "best" adherence and the other 4 groups were not statistically significant. In this observational study, treatment intensification was associated with similar BP improvement regardless of the patient's level of adherence. A randomized trial could further examine optimal management of patients with suboptimal adherence.


Establishing the European diagnostic reference levels for interventional cardiology.
Interventional cardiac procedures may be associated with high patient doses and therefore require special attention to protect the patients from radiation injuries such as skin erythema, cardiovascular tissue reactions or radiation-induced cancer. In this study, patient exposure data is collected from 13 countries (37 clinics and nearly 50 interventional rooms) and for 10 different procedures. Dose data was collected from a total of 14,922 interventional cardiology procedures. Based on these data European diagnostic reference levels (DRL) for air kerma-area product are suggested for coronary angiography (CA, DRL = 35 Gy cm2), percutaneous coronary intervention (PCI, 85 Gy cm2), transcatheter aortic valve implantation (TAVI, 130 Gy cm2), electrophysiological procedures (12 Gy cm2) and pacemaker implantations. Pacemaker implantations were further divided into single-chamber (2.5 Gy cm2) and dual chamber (3.5 Gy cm2) procedures and implantations of cardiac resynchronization therapy pacemaker (18 Gy cm2). Results show that relatively new techniques such as TAVI and treatment of chronic total occlusion (CTO) often produce relatively high doses, and thus emphasises the need for use of an optimization tool such as DRL to assist in reducing patient exposure. The generic DRL presented here facilitate comparison of patient exposure in interventional cardiology.


Identification of Borrelia burgdorferi ospC genotypes in canine tissue following tick infestation: implications for Lyme disease vaccine and diagnostic assay design.
In endemic regions, Lyme disease is a potential health threat to dogs. Canine Lyme disease manifests with arthritis-induced lameness, anorexia, fever, lethargy, lymphadenopathy and, in some cases, fatal glomerulonephritis. A recent study revealed that the regional mean for the percentage of seropositive dogs in the north-east of the USA is 11.6%. The outer surface protein C (OspC) of Lyme disease spirochetes is an important virulence factor required for the establishment of infection in mammals. It is a leading candidate in human and canine Lyme disease vaccine development efforts. Over 30 distinct ospC phyletic types have been defined. It has been hypothesized that ospC genotype may influence mammalian host range. In this study, Ixodes scapularis ticks collected from the field in Rhode Island were assessed for infection with B. burgdorferi. Ticks were fed on purpose bred beagles to repletion and infection of the dogs was assessed through serology and PCR. Tissue biopsies (n=2) were collected from each dog 49 days post-tick infestation (dpi) and the ospC genotype of the infecting strains determined by direct PCR of DNA extracted from tissue or by PCR after cultivation of spirochetes from biopsy samples. The dominant ospC types associated with B. burgdorferi canine infections differed from those associated with human infection, indicating a relationship between ospC sequence and preferred host range. Knowledge of the most common ospC genotypes associated specifically with infection of dogs will facilitate the rational design of OspC-based canine Lyme disease vaccines and diagnostic assays.


Anthropometric estimation of body height after surgical correction of scoliosis.
To estimate the height loss in 28 patients who had undergone surgery for scoliosis before skeletal maturity. 28 women patients aged 11 to 18 (mean, 15) years underwent Harrington instrumentation, segmental sublaminar wiring, and posterior spinal fusion for idiopathic scoliosis. The patients' mean age at menarche was 13 (range, 10-16) years. Pre- and post-operative radiographs of the spine were taken to determine curve types, the Cobb angle, and the degree of correction. The standing and sitting heights were measured using a flexible steel tape, and the expected standing and sitting heights were derived using formulae. At the final follow up, the mean patient age was 20 (16-32) years. The mean standing height was 155.2 cm and the mean sitting height was 77.4 cm. Using the formulae, the mean expected standing height was 158.4 cm, indicating height loss of 3.2 cm (t=4.6, p=0.0001), and the mean expected sitting height was 79.9 cm, indicating height loss of 2.6 cm (t=3.84, p=0.001). The curve types of the 28 patients were sub-divided as thoracic (n=7), thoracolumbar (n=18), and double major (n=3). Patients with thoracolumbar curves achieved better correction and less loss of standing and sitting heights. Percentage loss of standing and sitting heights were not correlated with the apex of the curve or the degree of correction. Height loss after surgery was acceptable in comparison with height loss associated with progression of scoliosis without surgery.


Family therapy for depression.
People with depression often experience interpersonal problems. Family therapy for depression is a widely used intervention, but it is unclear whether this is an effective therapy for the treatment of depression. To assess the efficacy of family therapy for depression. The following electronic databases were searched using a specific search strategy: CCDANCTR-Studies and CCDANCTR-References searched on 21/10/2005, The Cochrane Central Register of Controlled Trials, Medline (1966 to January 2005), EMBASE (1980 to January 2005), Psycinfo (1974 to January 2005). Reference lists of articles were also searched. Handsearches of relevant journals and bibliographies were conducted and first authors of included studies and experts in the field were contacted for further information. Included studies were randomised controlled and controlled clinical trials comparing family therapy with no intervention or an alternative intervention in which depression symptomatology was a main outcome measure. Methodological quality was independently assessed by two review authors using the Maastricht-Amsterdam Criteria List. The qualitative and quantitative characteristics of the selected trials were independently extracted by three review authors using a standardised data extraction form. Levels of evidence were used to determine the strength of the evidence available. It was not possible to perform meta-analyses because of the heterogeneity of the selected studies. Three high-quality and three low-quality studies, involving 519 people with depression, were identified. The studies were very heterogeneous in terms of interventions, participants, and measuring instruments. Despite fairly good methodological quality and positive findings of some studies, evidence for the effectiveness of family therapy for depression did not exceed level 3 (limited or conflicting evidence), except for moderate evidence (level 2), based on the non-combined findings from three studies, indicating that family therapy is more effective than no treatment or waiting list condition on decreasing depression, and on increasing family functioning. The current evidence base is too heterogeneous and sparse to draw conclusions on the overall effectiveness of family therapy in the treatment of depression. At this point, use of psychological interventions for the treatment of depression for which there is already an evidence-base would seem to be preferable to family therapy. Further high quality trials examining the effectiveness and comparative effectiveness of explicitly defined forms of family therapy are required.


Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury.
Insulin-like growth factor binding protein-2 (IGFBP-2) regulates the bioavailability, transportation, and localization of insulin-like growth factor-I (IGF-I), an effective neuroprotectant in animal stroke models especially when administered intranasally. Therefore, determining IGFBP-2's endogenous distribution in the normal and ischemic brain is essential in maximizing the neuroprotective potential of the intranasal IGF-I treatment approach. However, current data on IGFBP-2 is limited to mRNA and in situ hybridization studies. The purpose of this study was to determine if there are any changes in IGFBP-2 protein levels and distribution in ischemic brain and also to determine if IGFBPs play a role in the transportation of intranasally administered IGF-I into the brain. Using an in vitro approach, we show that ischemia causes changes in the distribution of IGFBP-2 in primary cortical neurons and astrocytes. In addition, we show using the transient middle cerebral artery occlusion (MCAO) model in mice that there is a significant increase in IGFBP-2 levels in the stroke penumbra and core after 72 h. This correlated with an overall increase in IGF-I after stroke, with the highest levels of IGF-I in the stroke core after 72 h. Brain sections from stroke mice indicate that neurons and astrocytes located in the penumbra both have increased expression of IGFBP-2, however, IGFBP-2 was not detected in microglia. We used binding competition studies to show that intranasally administered exogenous IGF-I uptake into the brain is not receptor mediated and is likely facilitated by IGFBPs. The change in protein levels indicates that IGFBP-2 plays an IGF-I-dependent and -independent role in the brain's acute (neuroprotection) and chronic (tissue remodeling) response to hypoxic-ischemic injury. Competition studies indicate that IGFBPs may have a role in rapid transportation of exogenous IGF-I from the nasal tissue to the site of injury.


From SARS to MERS, Thrusting Coronaviruses into the Spotlight.
Coronaviruses (CoVs) have formerly been regarded as relatively harmless respiratory pathogens to humans. However, two outbreaks of severe respiratory tract infection, caused by the severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome coronavirus (MERS-CoV), as a result of zoonotic CoVs crossing the species barrier, caused high pathogenicity and mortality rates in human populations. This brought CoVs global attention and highlighted the importance of controlling infectious pathogens at international borders. In this review, we focus on our current understanding of the epidemiology, pathogenesis, prevention, and treatment of SARS-CoV and MERS-CoV, as well as provides details on the pivotal structure and function of the spike proteins (S proteins) on the surface of each of these viruses. For building up more suitable animal models, we compare the current animal models recapitulating pathogenesis and summarize the potential role of host receptors contributing to diverse host affinity in various species. We outline the research still needed to fully elucidate the pathogenic mechanism of these viruses, to construct reproducible animal models, and ultimately develop countermeasures to conquer not only SARS-CoV and MERS-CoV, but also these emerging coronaviral diseases.


Avidity observed between a bivalent inhibitor and an enzyme monomer with a single active site.
Although myriad protein-protein interactions in nature use polyvalent binding, in which multiple ligands on one entity bind to multiple receptors on another, to date an affinity advantage of polyvalent binding has been demonstrated experimentally only in cases where the target receptor molecules are clustered prior to complex formation. Here, we demonstrate cooperativity in binding affinity (i.e., avidity) for a protein complex in which an engineered dimer of the amyloid precursor protein inhibitor (APPI), possessing two fully functional inhibitory loops, interacts with mesotrypsin, a soluble monomeric protein that does not self-associate or cluster spontaneously. We found that each inhibitory loop of the purified APPI homodimer was over three-fold more potent than the corresponding loop in the monovalent APPI inhibitor. This observation is consistent with a suggested mechanism whereby the two APPI loops in the homodimer simultaneously and reversibly bind two corresponding mesotrypsin monomers to mediate mesotrypsin dimerization. We propose a simple model for such dimerization that quantitatively explains the observed cooperativity in binding affinity. Binding cooperativity in this system reveals that the valency of ligands may affect avidity in protein-protein interactions including those of targets that are not surface-anchored and do not self-associate spontaneously. In this scenario, avidity may be explained by the enhanced concentration of ligand binding sites in proximity to the monomeric target, which may favor rebinding of the multiple ligand binding sites with the receptor molecules upon dissociation of the protein complex.


Mediating Effects of Positive Thinking and Social Support on Suicide Resilience.
Suicide has been the second leading cause of death for 18- to 24-year-olds in the United States since 2011. The stress experienced by undergraduate college students has the potential to increase one's risk for suicide. Resilience theory was used as a theoretical framework to examine the interplay between risk and protective factors. A cross-sectional and correlational design was used to assess the mediating effects of positive thinking and/or social support on suicide resilience in 131 college students 18 to 24 years old who completed an online survey. The study found an indirect effect of self-esteem on suicide resilience through positive thinking and social support indicating that as self-esteem increases, positive thinking and social support also increase, which leads to an increase in resilience. The study also found a direct effect of self-esteem, positive thinking, and social support on suicide resilience. The findings inform the development of tailored interventions to build suicide resilience in college students.


Multiple roles of metal ions in large ribozymes.
Since the discovery of catalytic RNA molecules (ribozymes), intense research has been devoted to understand their structure and activity. Among RNA molecules, the large ribozymes, namely group I and group II introns and RNase P, are of special importance. The first two ribozymes are known for their ability to perform self-splicing while RNase P is responsible for the 5'-end maturation of tRNA in bacteria, archea, and eukaryotes. All three groups of ribozymes show a significant requirement for metal ions in order to establish the active tertiary structure that enables catalysis. The primary role of both monovalent and divalent metal ions is to screen the negative charge associated with the phosphate sugar backbone, but the metal ions also play an active role in catalysis. Biochemical and biophysical investigations, supported by recent findings from X-ray crystal structures, allow clarifying and rationalizing both the structural and catalytic roles of metal ions in large ribozymes. In particular, the "two-metal-ion mechanism", describing how metal ions in the active center take part in catalysis, has been largely corroborated.


Characterization of a small (25-kilodalton) derivative of the Rous sarcoma virus Gag protein competent for particle release.
Retroviral Gag proteins have the ability to induce budding and particle release from the plasma membrane when expressed in the absence of all of the other virus-encoded components; however, the locations of the functional domains within the Gag protein that are important for this process are poorly understood. It was shown previously that the protease sequence of the Rous sarcoma virus (RSV) Gag protein can be replaced with a foreign polypeptide, iso-1-cytochrome c from a yeast, without disrupting particle assembly (R. A. Weldon, Jr., C. R. Erdie, M. G. Oliver, and J. W. Wills, J. Virol. 64:4169-4179, 1990). An unexpected product of the chimeric gag gene is a small, Gag-related protein named p25C. This product was of interest because of its high efficiency of packaging into particles. The goal of the experiments described here was to determine the mechanism by which p25C is synthesized and packaged into particles. The results demonstrate that it is not the product of proteolytic processing of the Gag-cytochrome precursor but is derived from an unusual spliced mRNA. cDNA clones of the spliced mRNA were obtained, and each expressed a product of approximately 25 kDa, designated p25M1, which was released into the growth medium in membrane-enclosed particles that were much lighter than authentic retrovirions as measured in sucrose density gradients. DNA sequencing revealed that the clones encode the first 180 of the 701 amino acids of the RSV Gag protein and no residues from iso-1-cytochrome c. This suggested that a domain in the carboxy-terminal half of Gag is important for the packaging of Gag proteins into dense arrays within the particles. In support of this hypothesis, particles of the correct density were obtained when a small segment from the carboxy terminus of the RSV Gag protein (residues 417 to 584) was included on the end of p25.


Characterization of the archaeal thermophile Sulfolobus turreted icosahedral virus validates an evolutionary link among double-stranded DNA viruses from all domains of life.
Icosahedral nontailed double-stranded DNA (dsDNA) viruses are present in all three domains of life, leading to speculation about a common viral ancestor that predates the divergence of Eukarya, Bacteria, and Archaea. This suggestion is supported by the shared general architecture of this group of viruses and the common fold of their major capsid protein. However, limited information on the diversity and replication of archaeal viruses, in general, has hampered further analysis. Sulfolobus turreted icosahedral virus (STIV), isolated from a hot spring in Yellowstone National Park, was the first icosahedral virus with an archaeal host to be described. Here we present a detailed characterization of the components forming this unusual virus. Using a proteomics-based approach, we identified nine viral and two host proteins from purified STIV particles. Interestingly, one of the viral proteins originates from a reading frame lacking a consensus start site. The major capsid protein (B345) was found to be glycosylated, implying a strong similarity to proteins from other dsDNA viruses. Sequence analysis and structural predication of virion-associated viral proteins suggest that they may have roles in DNA packaging, penton formation, and protein-protein interaction. The presence of an internal lipid layer containing acidic tetraether lipids has also been confirmed. The previously presented structural models in conjunction with the protein, lipid, and carbohydrate information reported here reveal that STIV is strikingly similar to viruses associated with the Bacteria and Eukarya domains of life, further strengthening the hypothesis for a common ancestor of this group of dsDNA viruses from all domains of life.


Effect of a Soft Robotic Sock Device on Lower Extremity Rehabilitation Following Stroke: A Preliminary Clinical Study With Focus on Deep Vein Thrombosis Prevention.
Immobility of the lower extremity due to medical conditions such as stroke can lead to medical complications such as deep vein thrombosis or ankle contracture, and thereafter prolonged recovery process of the patients. In this preliminary clinical study, we aimed to examine the effect of a novel soft robotic sock device, capable of providing assisted ankle exercise, in improving blood flow in the lower limb to prevent the complication of strokes such as deep vein thrombosis and joint contracture. Stroke patients were recruited (n = 17) to compare patients using the conventional pneumatic compression device with our robotic sock device on separate days. The primary outcome was to compare the venous flow profile of the superficial femoral vein in terms of the time average mean velocity and volumetric flow. The secondary outcome was to identify the ankle joint range of motion with the assistance of the device. We noted improvements in the venous profile at the early phase of the device use, though its efficacy seemed to drop with time, as compared to the IPC device, where there was a significant improvement in the venous profile. The ankle joint dorsiflexion-plantarflexion range of motion assisted by the device was 11.5±6.3°. Conclusion and clinical impact: The current version of our sock device appears to be capable of improving venous blood flow in the early phase of device use and assisting with ankle joint exercise. The insights from this preliminary clinical study will serve as the basis for further improvement of the device and subsequent conduct of a longitudinal clinical trial. National Health Innovation Centre Singapore (NHIC) grant, R-172-000-391-511, MOE AcRF Tier 1 R-397-000-301-114.


[Hepatitis B virus morphogenesis].
The human hepatitis B virus (HBV) is a small hepatotropic enveloped virus associated with chronic infection that can lead to cirrhosis and hepatocellular carcinoma. The HBV genome is a DNA molecule contained in an icosahedral capsid. Although HBV is not a retrovirus, the replication of its genome involves reverse transcription. Another distinctive feature of HBV is the production, in great excess over virions, of non-infectious subviral particles (SVP) consisting of membrane phospholipids and the three envelope proteins (small [S], medium [M] and large [L]). These empty non-infectious particles are highly immunogenic, and their in vitro production is at the basis of the current vaccine against hepatitis B. Despite numerous studies that lead to a better understanding of the HBV replication, little is known about the morphogenesis of the virion and its associated SVP. Recent approaches suggest that the mechanisms responsable for assembly of the virions and the SVP could be distinct.


Indirect inhibitory effect of a neurotensin receptor antagonist on human colon cancer (LoVo) growth.
The effect of the nonpeptide neurotensin (NT) receptor antagonist SR48692 on NT-mediated growth of xenografted human colon cancers (LoVo) was determined. Sixty-four athymic nude mice, inoculated with LoVo cells at a single site, were randomized into four groups of 16 mice each to receive either vehicle NT (600 microg/kg), SR48692 (2 mg/kg) or NT+SR48692 administered s.c., t.i.d., for 25 days. Treatment with NT significantly stimulated LoVo tumour growth, weight, DNA and protein content; SR48692 blocked this NT-mediated effect but had no effect when administered as a single agent. In addition, normal jejunum and ileum were removed and assessed. Similar to the effects on LoVo tumours, NT stimulated jejunal and ileal growth; SR48692 blocked this NT-mediated effect. In contrast to our in vivo findings, NT had no effect on LoVo cell growth in vitro. Also, Northern blot analysis demonstrated no expression for the NT receptor in either LoVo tumour cells or xenografted tumours. Our findings suggest that the trophic effect of NT on LoVo may be through an indirect effect; one possibility is that administration of NT may stimulate the release of other trophic factors which then stimulate tumour growth. The nonpeptide NT receptor antagonist SR48692 will be a useful agent to delineate the specific effects of NT on neoplastic as well as normal gastrointestinal tissues.


Epidemiologic evidence for multiple sclerosis as an infection.
The worldwide distribution of multiple sclerosis (MS) can be described within three zones of frequency: high, medium, and low. The disease has a predilection for white races and for women. Migration studies show that changing residence changes MS risk. Studies of persons moving from high- to low-risk areas indicate that in the high-risk areas, MS is acquired by about age 15. Moves from low- to high-risk areas suggest that susceptibility is limited to persons between about ages 11 and 45. MS on the Faroe Islands has occurred as four successive epidemics beginning in 1943. The disease appears to have been introduced by British troops who occupied the islands for 5 years from 1940, and it has remained geographically localized within the Faroes for half a century. What was introduced must have been an infection, called the primary MS affection (PMSA), that was spread to and from successive cohorts of Faroese. In this concept, PMSA is a single widespread systemic infectious disease (perhaps asymptomatic) that only seldom leads to clinical neurologic MS. PMSA is also characterized by a need for prolonged exposure, limited age of susceptibility, and prolonged incubation. I believe that clinical MS is the rare late outcome of a specific, but unknown, infectious disease of adolescence and young adulthood and that this infection could well be caused by a thus-far-unidentified (retro)virus.


An epidemiological study of osteogenic sarcoma in Kenya: the variations in incidence between ethnic groups and geographic regions, 1968-1978.
Kenya is a country of marked environmental and ethnic diversity. A study of osteogenic sarcoma occurring in Kenya from 1968 to 1978 revealed 251 cases, representing between 89% and 100% of the predicted number. Variations in age, sex and anatomical location were within classical limits. However, the incidence of osteogenic sarcoma amongst the Central Bantu was significantly higher than predicted (P less than 0.0001), whilst the incidence among the Western Bantu was significantly lower (P less than 0.002), despite their similar ethnic origins. Two geographically dissimilar areas likewise exhibited significant differences in incidence. The Eastern province showed a higher incidence (P less than 0.02), whereas the Nyanza Province (P less than 0.001) and the adjacent Western Province (P less than 0.005) showed a lower than predicted incidence. These observations suggest that in Kenya a geomedical variable affects the incidence of osteogenic sarcoma and that genetic variation has no effect on incidence.


A SWI/SNF Chromatin Remodelling Protein Controls Cytokinin Production through the Regulation of Chromatin Architecture.
Chromatin architecture determines transcriptional accessibility to DNA and consequently gene expression levels in response to developmental and environmental stimuli. Recently, chromatin remodelers such as SWI/SNF complexes have been recognized as key regulators of chromatin architecture. To gain insight into the function of these complexes during root development, we have analyzed Arabidopsis knock-down lines for one sub-unit of SWI/SNF complexes: BAF60. Here, we show that BAF60 is a positive regulator of root development and cell cycle progression in the root meristem via its ability to down-regulate cytokinin production. By opposing both the deposition of active histone marks and the formation of a chromatin regulatory loop, BAF60 negatively regulates two crucial target genes for cytokinin biosynthesis (IPT3 and IPT7) and one cell cycle inhibitor (KRP7). Our results demonstrate that SWI/SNF complexes containing BAF60 are key factors governing the equilibrium between formation and dissociation of a chromatin loop controlling phytohormone production and cell cycle progression.


In vitro propagation of primary and extended life span murine corneal endothelial cells.
To establish primary and extended life span cell cultures of murine corneal endothelial cells for a model system investigating corneal endothelial cell replacement and the immunologic features of corneal graft rejection. The authors have been able to grow corneal endothelium in culture by isolating adult murine Descemet's membrane and allowing the endothelial cells to proliferate from the explants. To isolate extended life span murine corneal endothelial cells, cells were infected with an adenovirus-SV40 hybrid virus (Ad12-SV40). The primary cells from adult corneas proliferated to passage 3 before growth arrest-senescence was observed. However, the extended life span cells remained proliferative through passage 36 and maintained a structure similar to the primary cells. Immunohistochemical analysis demonstrate that the extended life span cells are SV40 large T antigen positive, and both the primary and extended life span murine corneal endothelial cells exhibit the common expression of several growth factor receptors and TGF-beta. This is the first report of the isolation and culture of mouse corneal endothelial cells. Additionally, these cells have been infected with a virus carrying the SV40 large T antigen, which yields extended life span mouse corneal endothelial cells. These cells will be of interest in establishing a murine model for corneal cell transplantation, and the authors are currently establishing protocols for the specific introduction and manipulation of these cells in both in vitro and in vivo systems. These types of analyses may provide an important animal model specific to in vivo corneal endothelial cell replacement for the treatment of endothelial-related keratopathies and can be a useful model in delineating the immunologic parameters of corneal graft rejection.


Relationships of objectively scored Bender variables with MMPI scores in an outpatient psychiatric population.
The purpose of the present study was to examine whether the Advanced Psychodiagnostic Interpretation system for the Bender Gestalt Test could reasonably predict the results of the most widely used objective measure of personality, the MMPI. Despite the widespread use of both tests, no previous studies could be found which correlated actual Bender scores with MMPI results, arising partly from the lack of a well-accepted, reliable, and objective scoring system for the Bender. The study compared the performance of 279 adult psychological outpatients on both the MMPI and Bender. The 55 Bender scorable points, which are seen most frequently in the outpatient population, were factor analyzed to yield 17 factors which were correlated with the MMPI. Significant multiple correlations were found between the Bender factors and 10 of 12 MMPI scales, with significant correlations ranging from .36 to .47. The Bender overall was able to discriminate moderately high scorers on the MMPI from low scorers. The overall results suggested that the Advanced Psychodiagnostic Interpretation scoring system includes measures that reflect general psychopathology and correlate with the MMPI as well as more specific content that is independent of the MMPI scales. The potential of this scoring system and joint use of the MMPI and Bender in personality assessment are discussed. Replication with a larger sample than 279 is encouraged for these 55 Bender and 12 MMPI items.


Rainbow Trout Maintain Intestinal Transport and Barrier Functions Following Exposure to Polystyrene Microplastics.
Ingestion has been proposed as a prominent exposure route for plastic debris in aquatic organisms, including fish. While the consequences of ingestion of large plastic litter are mostly understood, the impacts resulting from ingestion of microplastics (MPs) are largely unknown. We designed a study that aimed to assess impacts of MPs on fish intestinal physiology and examined integrity of extrinsic, physical and immunological barriers. Rainbow trout were exposed to polystyrene (PS) MPs (100-400 μm) via feed for a period of 4 weeks. Fish were fed four types of diets: control, diets containing virgin PS particles, or particles exposed to two different environmental matrices (sewage or harbor effluent). Extrinsic barrier disturbance in intestinal tissue was evaluated via histology. The paracellular permeability toward ions and molecules was examined using Ussing chambers and mRNA expression analysis of tight junction proteins. Active transport was monitored as transepithelial potential difference, short-circuits current and uptake rate of amino acid 3H-lysine. Immune status parameters were measured through mRNA expression level of cytokines, lysozyme activity, and hematological analysis of immune cells. We could not show that PS MPs induced inflammatory responses or acted as physical or chemical hazards upon ingestion. No measurable effects were exerted on fish intestinal permeability, active transport or electrophysiology.


Protein family expansions and biological complexity.
During the course of evolution, new proteins are produced very largely as the result of gene duplication, divergence and, in many cases, combination. This means that proteins or protein domains belong to families or, in cases where their relationships can only be recognised on the basis of structure, superfamilies whose members descended from a common ancestor. The size of superfamilies can vary greatly. Also, during the course of evolution organisms of increasing complexity have arisen. In this paper we determine the identity of those superfamilies whose relative sizes in different organisms are highly correlated to the complexity of the organisms. As a measure of the complexity of 38 uni- and multicellular eukaryotes we took the number of different cell types of which they are composed. Of 1,219 superfamilies, there are 194 whose sizes in the 38 organisms are strongly correlated with the number of cell types in the organisms. We give outline descriptions of these superfamilies. Half are involved in extracellular processes or regulation and smaller proportions in other types of activity. Half of all superfamilies have no significant correlation with complexity. We also determined whether the expansions of large superfamilies correlate with each other. We found three large clusters of correlated expansions: one involves expansions in both vertebrates and plants, one just in vertebrates, and one just in plants. Our work identifies important protein families and provides one explanation of the discrepancy between the total number of genes and the apparent physiological complexity of eukaryotic organisms.


Central nervous system abnormalities in fibromyalgia and chronic fatigue syndrome: new concepts in treatment.
Fibromyalgia (FM) and chronic fatigue syndrome (CFS) are poorly understood disorders that share similar demographic and clinical characteristics. The etiology and pathophysiology of these diseases remain unclear. Because of the similarities between both disorders it was suggested that they share a common pathophysiological mechanisms, namely, central nervous system (CNS) dysfunction. Current hypotheses center on atypical sensory processing in the CNS and dysfunction of skeletal muscle nociception and the hypothalamic-pituitary-adrenal (HPA) axis. Researches suggest that the (CNS) is primarily involved in both disorders in regard to the pain, fatigue and sleep disturbances. Many patients experience difficulty with concentration and memory and many others have mood disturbance, including depression and anxiety. Although fibromyalgia is common and associated with substantial morbidity and disability, there are no US Food and Drug Administration (FDA)-approved treatments except pregabalin. Recent pharmacological treatment studies about fibromyalgia have focused on selective serotonin and norepinephrine (NE) reuptake inhibitors, which enhance serotonin and NE neurotransmission in the descending pain pathways and lack many of the adverse side effects associated with tricyclic medications. CFS is a descriptive term used to define a recognisable pattern of symptoms that cannot be attributed to any alternative condition. The symptoms are currently believed to be the result of disturbed brain function. To date, no pharmacological agent has been reliably shown to be effective treatment for CFS. Management strategies are therefore primarily directed at relief of symptoms and minimising impediments to recovery. This chapter presents data demonstrating CFS, abnormal pain processing and autonomic nervous system (ANS) dysfunction in FM and CFS and concludes by reviewing the new concepts in treatments in CFS and FM.


siAbasic: a comprehensive database for potent siRNA-6Ø sequences without off-target effects.
Small interfering RNA (siRNA) is widely used to specifically silence target gene expression, but its microRNA (miRNA)-like function inevitably suppresses hundreds of off-targets. Recently, complete elimination of the off-target repression has been achieved by introducing an abasic nucleotide to the pivot (position 6; siRNA-6Ø), of which impaired base pairing destabilizes transitional nucleation (positions 2-6). However, siRNA-6Ø varied in its conservation of on-target activity (∼80-100%), demanding bioinformatics to discover the principles underlying its on-target efficiency. Analyses of miRNA-target interactions (Ago HITS-CLIP) showed that the stability of transitional nucleation correlated with the target affinity of RNA interference. Furthermore, interrogated analyses of siRNA screening efficiency, experimental data and broadly conserved miRNA sequences showed that the free energy of transitional nucleation (positions 2-5) in siRNA-6Ø required the range of stability for effective on-target activity (-6 ≤ ΔG[2:5] ≤ -3.5 kcal mol-1). Taking into consideration of these features together with locations, guanine-cytosine content (GC content), nucleotide stretches, single nucleotide polymorphisms and repetitive elements, we implemented a database named 'siAbasic' that provided the list of potent siRNA-6Ø sequences for most of human and mouse genes (≥ ∼95%), wherein we experimentally validated some of their therapeutic potency. siAbasic will aid to ensure potency of siRNA-6Ø sequences without concerning off-target effects for experimental and clinical purposes.


Chronic Fluoxetine Treatment Induces Maturation-Compatible Changes in the Dendritic Arbor and in Synaptic Responses in the Auditory Cortex.
Fluoxetine is a selective serotonin reuptake inhibitor (SSRI) used to treat mood and anxiety disorders. Chronic treatment with this antidepressant drug is thought to favor functional recovery by promoting structural and molecular changes in several forebrain areas. At the synaptic level, chronic fluoxetine induces an increased size and density of dendritic spines and an increased ratio of GluN2A over GluN2B N-methyl-D-aspartate (NMDA) receptor subunits. The "maturation"-promoting molecular changes observed after chronic fluoxetine should also induce structural remodeling of the neuronal dendritic arbor and changes in the synaptic responses. We treated adult rats with fluoxetine (0.7 mg/kg i.p. for 28 days) and performed a morphometric analysis using Golgi stain in limbic and nonlimbic cortical areas. Then, we focused especially on the auditory cortex, where we evaluated the dendritic morphology of pyramidal neurons using a 3-dimensional reconstruction of neurons expressing mRFP after in utero electroporation. With both methodologies, a shortening and decreased complexity of the dendritic arbors was observed, which is compatible with an increased GluN2A over GluN2B ratio. Recordings of extracellular excitatory postsynaptic potentials in the auditory cortex revealed an increased synaptic response after fluoxetine and were consistent with an enrichment of GluN2A-containing NMDA receptors. Our results confirm that fluoxetine favors maturation and refinement of extensive cortical networks, including the auditory cortex. The fluoxetine-induced receptor switch may decrease GluN2B-dependent toxicity and thus could be applied in the future to treat neurodegenerative brain disorders characterized by glutamate toxicity and/or by an aberrant network connectivity.


Decline of photopic multifocal electroretinogram responses with age is due primarily to preretinal optical factors.
Age-related changes in photopic retinal function were evaluated topographically with the multifocal electroretinogram (mfERG). Thirty-two subjects between the ages of 16 and 69 participated. There was a strong dependence on age for all mfERG response measures that was strongest for the group of central retinal responses (i.e., within 5 deg eccentricity) and approximately equal for responses between 5 and 20 deg. After adjustment for crystalline lens optical density and pupil diameter, significant effects of age were limited to central first-order (i.e., within 5 deg) and second-order response kernels. Simulation studies support an optical basis for the observed age-related changes. It is concluded that mfERG changes between the ages of 20 and 70 are due predominantly to preretinal optical factors.


Biodegradable Nanocomposite with Dual Cell-Tissue Penetration for Deep Tumor Chemo-Phototherapy.
Chemo-phototherapy, as a promising cancer combination therapy strategy, has attracted widespread attention. However, the complex tumor microenvironment restricts the penetration depth of chemo-phototherapy agents in the tumor region. Here, biodegradable amphiphilic gelatin (AG) wrapped nanocomposite (PRDCuS@AG) composed of doxorubicin and copper sulfide (CuS)-loaded dendrimer is designed for deep tumor chemo-phototherapy. PR in PRDCuS@AG represents arginine-conjugated polyamidoamine dendrimer. PRDCuS@AG can rapidly biodegrade into PRDCuS by matrix metalloproteinases under near-infrared light irradiation. The resulted PRDCuS harbors dual cell-tissue penetration ability, which can effectively penetrate deep into the tumor tissue. In particular, PRDCuS@AG achieves photoacoustic imaging-guided synergistic chemo-phototherapy with 97% of tumor inhibition rate. Moreover, PRDCuS@AG can further degrade into 3 nm ultrasmall CuS, which can be eliminated from the body after treatment to avoid side effects. This strategy provides an insight that the development of chemo-phototherapy agents with high penetration ability to overcome the limitation of current deep tumor therapy.