Prognosis of undiagnosed chest pain: linked electronic health record cohort study.
Objective To ascertain long term cardiovascular outcomes in patients whose chest pain remained undiagnosed six months after first presentation.Design Cohort study.Setting UK electronic health record database (CALIBER) linking primary care, secondary care, coronary registry, and death registry information.Participants 172 180 adults aged ≥18 from 223 general practices presenting with a first episode of recorded chest pain, classified from medical records as diagnosed (non-coronary condition or angina) or undiagnosed (cause unattributed) at first consultation between 2002 and 2009 and with no previous record of cardiovascular disease.Main outcome measures Fatal or non-fatal cardiovascular events over 5.5 years' follow-up. Adjustments were made for age, sex, deprivation, body mass index, smoking status, year of index presentation, and previous records of diabetes or hypertension or previous prescriptions for lipid lowering drugs.Results At the index presentation, 72.4% of patients (124 688) did not have a cause attributed for their chest pain; 118 687 (95.2%) of these did not receive any type of cardiovascular diagnosis over the next six months. Only a minority of patients in all three groups (non-coronary 2.0% (769 of 39 232); unattributed 11.7% (14 582 of 124 688); angina 31.5% (2606 of 8260)) had a recorded cardiac diagnostic investigation in the first six months after presentation. The long term incidence of cardiovascular events was higher in those whose chest pain remained unattributed after six months (5126 of 109 628; 4.7%) compared with patients with an initial diagnosis of non-coronary pain (1073 of 36 097; 3.0%) (adjusted hazard ratios for 0.5-1 year after presentation: 1.95, 95% confidence interval 1.66 to 2.31; for 1-3 years: 1.35, 1.23 to 1.48); for 3-5.5 years: 1.21, 1.08 to 1.37). Owing to the larger number of patients in the unattributed group, there were more excess myocardial infarctions in the long term in this group (214 more than expected based on the rate in the non-coronary group) than in the angina group (132 more than expected). Patients who had cardiac diagnostic investigations in the first six months had a higher long term risk of cardiovascular events, regardless of the initial chest pain label. Incidence of unattributed chest pain and angina decreased between 2002 (124 per 10 000 person years and 13 per 10 000 person years, respectively) and 2009 (107 per 10 000 person years and 5 per 10 000 person years, respectively), but the incidence of chest pain attributed to a non-coronary cause remained stable (37-40 per 10 000 person years). Risk of cardiovascular events did not change over time.Conclusions Most patients with first onset chest pain do not have a diagnosis recorded at presentation or in the subsequent six months, including those who undergo cardiac investigations. These patients have an increased risk of cardiovascular events for at least five years. Efforts to better assess and reduce the cardiovascular risk of such patients are warranted.


Local linear estimation for spatial random processes with stochastic trend and stationary noise.
We consider the problem of estimating the trend for a spatial random process model expressed as Z(x) = μ(x) + ε(x) + δ(x), where the trend μ is a smooth random function, ε(x) is a mean zero, stationary random process, and {δ(x)} are assumed to be i.i.d. noise with zero mean. We propose a new model for stochastic trend in ℝ d by generalizing the notion of a structural model for trend in time series. We estimate the stochastic trend nonparametrically using a local linear regression method and derive the asymptotic mean squared error of the trend estimate under the proposed model for trend. Our results show that the asymptotic mean squared error for the stochastic trend is of the same order of magnitude as that of a deterministic trend of comparable complexity. This result suggests from the point of view of estimation under stationary noise, it is immaterial whether the trend is treated as deterministic or stochastic. Moreover, we show that the rate of convergence of the estimator is determined by the degree of decay of the correlation function of the stationary process ε(x) and this rate can be different from the usual rate of convergence found in the literature on nonparametric function estimation. We also propose a data dependent selection procedure for the bandwidth parameter which is based on a generalization of Mallow's C p criterion. We illustrate the methodology by simulation studies and by analyzing a data on surface temperature anomalies.


Modified spectrum autointerferometric correlation (MOSAIC) for single-shot pulse characterization.
A method for generation of the modified spectrum autointerferometric correlation that allows single-shot pulse characterization is demonstrated. A sensitive graphical representation of the ultrashort pulse phase quality is introduced that delineates the difference between the presence of temporal and spectral phase distortions. Using these schemes, full-field reconstruction of ultrashort laser pulses is obtained in real time using an efficient iterative technique.


Retail-based healthy food point-of-decision prompts (PDPs) increase healthy food choices in a rural, low-income, minority community.
This study examines the potential for point-of-decision prompts (PDPs) to promote healthier food choices among shoppers in a rural, low-income, minority community. We hypothesized that a narrowly defined PDP (focused on fresh produce) would be easier for shoppers to remember than a broadly defined PDP (focused on any healthy items), resulting in a higher proportion of healthy items purchased. PDPs were placed at the entrance to a supermarket in Mission, South Dakota, United States of America, on the Rosebud Sioux Reservation for alternating time periods, July 9-10, 2017. Sales records from 653 transactions were retrieved from the supermarket, comprising periods in which PDPs were in place and control periods. We examined the proportion of selected items and proportion of total expenditures that were a) any healthy foods and b) fresh fruits and vegetables. Data were analyzed in 2018. The narrowly defined prompt consistently resulted in a higher proportion of items and expenditures on healthy foods than either the broad prompt or the control condition. Shoppers in the narrow prompt condition purchased and spent significantly more on any healthy foods and fresh produce than shoppers in the control condition. While shoppers in the narrow prompt condition purchased more healthy foods and fresh produce than shoppers in the broad prompt condition, the differences were not statistically significant. Shoppers exposed to the narrow PDP consistently purchased more healthy foods than shoppers in a control group, while shoppers in the broad PDP did not, highlighting the importance of considering cognitive processes when designing health promotion messages.


Electrophysiology and dye-coupling are sexually dimorphic characteristics of individual laryngeal muscle fibers in Xenopus laevis.
Sex differences at the laryngeal neuromuscular junction of Xenopus laevis were examined by recording intracellularly from muscle fibers in response to nerve stimulation. Male laryngeal muscle contains 2 physiologically distinct fiber types. Type I fibers generate postsynaptic potentials in response to low-magnitude stimulus pulses and action potentials in response to higher-magnitude stimulus pulses. Type II muscle fibers require repetitive stimulation for action potential production, probably because of facilitation. Subthreshold events in type I and II fibers suggest that these neuromuscular synapses have low safety factor junctions. Female laryngeal muscle contains one fiber type (III), which is physiologically distinct from those found in the male. Type III fibers produce an action potential in response to a single-stimulus pulse of suprathreshold voltage delivered to the laryngeal nerve; subthreshold events were not observed. Iontophoretic injection of Lucifer yellow into a single female muscle fiber resulted in as many as 43 labeled fibers. In males, only one fiber was labeled. Dye-coupling was not observed in adult females treated with the androgenic steroid hormone, testosterone. We have previously reported that laryngeal muscle fibers are recruited throughout a stimulus train presented to the laryngeal nerve in males, but are not recruited in females (Tobias and Kelly, 1987). Sex differences in the frequency of electrophysiological fiber types described here may account for sex differences in fiber recruitment. Synchronous activity of dye-coupled fibers may increase the effectiveness of muscle contraction in females.


Narrative Interaction in Family Dinnertime Conversations.
Reminiscing has been shown to be a critical conversational context for the development of autobiographical memory, self-concept, and emotional regulation (for a review, see Fivush, Haden, & Reese, 2006). Although much past research has examined reminiscing between mothers and their preschool children, very little attention has been given to family narrative interaction with older children. In the present study, we examined family reminiscing in spontaneous narratives that emerged during family dinnertime conversations. The results revealed that mothers contributed more to the narratives than did fathers in that they provided, confirmed, and negated more information, although fathers requested more information than mothers. In exploratory analyses, mothers' contributions to shared family narratives were found to be related to fewer internalizing and externalizing behaviors in their children, while fathers' contributions to individual narratives of day-today experiences were related to fewer internalizing and externalizing behaviors in their children. These results indicate that mothers and fathers may play different roles in narrative construction with their children, and there is some suggestion that these differences may also be related to children's behavioral adjustment.


Gallamine (Flaxedil) and synaptic transmission in the spinal cord.
A paralyzing dose of gallamine (Flaxedil) (1 to 2 milligrams per kilogram of body weight) has no effect on synaptic transmission in the cat's spinal cord. In spinal cats ventilated with oxygen, we stimulated a dorsal spinal root and recorded the compound ventral root potential. The reflex potential was not affected by 6.25 milligrams of gallamine per kilogram. Giving 12.5 milligrams of gallamine per kilogram had no significant effect on the monosynaptic spike height, but the polysynaptic response rose briefly to 12 percent above control. Increased magnitude of the polysynaptic response appeared related to a concomitant rise in blood pressure.


Adverse events following HPV vaccination: 11 years of surveillance in Australia.
Australia was the first country to implement a fully funded vaccination program with quadrivalent human papillomavirus vaccine (4vHPV) in 2007, including males from 2013. We examined adverse events (AE) following vaccination with 4vHPV from 11 years of post-marketing data, focusing on a period of enhanced surveillance and adverse events of special interest (AESI). AE following 4vHPV doses administered between April 2007 and December 2017 reported to Australia's national regulator, the Therapeutic Goods Administration, were examined; reports collected during enhanced surveillance in 2013 and 2014 were analyzed separately. Age and sex-specific rates, using denominator data from the national HPV vaccination register, were determined. Pre-specified AESI were identified using Medical Dictionary for Regulatory Activities (MedDRA®) Preferred Terms and examined in detail. Following nine million doses of 4vHPV vaccine administered in Australia, 4551 AE reports were identified. The crude reporting rate was 39.8 per 100 000 doses in the funded cohorts, excluding the enhanced surveillance period. The reported rate of syncope in 12 to 13-year-old males and females was 29.6 per 100 000 doses during enhanced surveillance and 7.1 per 100 000 doses during the remaining study period; rates of syncope were higher in younger compared to older adolescents. The rate of anaphylaxis (0.32 per 100 000 doses) was consistent with published rates. Other AESI including autoimmune disease, postural orthostatic tachycardia syndrome, primary ovarian insufficiency, Guillain-Barré syndrome, complex regional pain syndrome and venous thromboembolism, were reported at low rates and analysis did not reveal unexpected patterns that would suggest causal association. AESI, apart from syncope, were reported rarely. The higher rate of syncope among younger adolescents highlights the need for management protocols to prevent syncope-related injury. Analysis of this large, longitudinal dataset in a country with high vaccine uptake, including a period of enhanced surveillance, affirms the safety profile of 4vHPV.


Targeting fibroblast growth factor pathways in prostate cancer.
Advanced prostate cancer carries a poor prognosis and novel therapies are needed. Research has focused on identifying mechanisms that promote angiogenesis and cellular proliferation during prostate cancer progression from the primary tumor to bone-the principal site of prostate cancer metastases. One candidate pathway is the fibroblast growth factor (FGF) axis. Aberrant expression of FGF ligands and FGF receptors leads to constitutive activation of multiple downstream pathways involved in prostate cancer progression including mitogen-activated protein kinase, phosphoinositide 3-kinase, and phospholipase Cγ. The involvement of FGF pathways in multiple mechanisms relevant to prostate tumorigenesis provides a rationale for the therapeutic blockade of this pathway, and two small-molecule tyrosine kinase inhibitors-dovitinib and nintedanib-are currently in phase II clinical development for advanced prostate cancer. Preliminary results from these trials suggest that FGF pathway inhibition represents a promising new strategy to treat castrate-resistant disease.


Heterotrimeric G-proteins are implicated in the regulation of apoptosis in pancreatic beta-cells.
Recent studies have provided evidence that apoptosis of pancreatic beta-cells is important in the early etiology of both type I and type II diabetes mellitus. The mechanisms responsible for induction of apoptosis are unknown, but we present evidence that the signal transduction pathway controlling the process in pancreatic beta-cells is regulated by G-proteins. We have employed the global G-protein activator fluoride and show that this agent induces apoptosis in clonal RINm5F pancreatic beta-cells and also in the cells of normal rat islets of Langerhans. The process is time and concentration dependent and may reflect the formation of AIF4- since it was inhibited by the aluminum chelator deferoxamine. Induction of apoptosis by fluoride was confirmed by acridine orange staining of cell nuclei, by electron-microscopic examination of chromatin condensation, and by oligonucleosomal degradation of DNA. The involvement of G-proteins was confirmed by culture of beta-cells in the presence of pertussis toxin (PTX) prior to exposure to fluoride. PTX did not affect the extent of cell death under control conditions but it consistently, and markedly, enhanced the response to fluoride. The results demonstrate that apoptosis can be induced in pancreatic beta-cells by sustained activation of a G-protein-dependent signaling pathway(s) and they further suggest that a pertussis toxin-sensitive G-protein is involved in attenuation of the response. Treatment of RINm5F pancreatic beta-cells with dibutyrylcAMP resulted in a dose-dependent, saturable increase in cell death, suggesting that a sustained rise in intracellular cAMP may form part of the effector system controlling apoptosis.


A reactive coordination scheme for a many-robot system.
This paper presents a novel approach for coordinating a homogeneous system of mobile robots using implicit communication in the form of broadcasts. The broadcast-based coordination scheme was developed for the Army Ant swarm-a system of small, relatively inexpensive mobile robots that can accomplish complex tasks by cooperating as a team. The primary drawback, however, of the Army Ant system is that the absence of a central supervisor poses difficulty in the coordination and control of the agents. Our coordination scheme provides a global "group dynamic" that controls the actions of each robot using only local interactions. Coordination of the swarm is achieved with signals we call "heartbeats". Each agent broadcasts a unique heartbeat and responds to the collective behavior of all other heartbeats. We generate heartbeats with van der Pol oscillators. In this application, we use the known properties of coupled van der Pol oscillators to create predictable group behavior. Some of the properties and behaviors of coupled van der Pol oscillators are discussed in detail. We emphasize the use of this scheme to allow agents to simultaneously perform an action such as lifting, steering, or changing speed. The results of experiments performed on three actual heartbeat circuits are presented and the behavior of the realized system is compared to simulated results. We also demonstrate the application of the coordination scheme to global speed control.


Peripheral blood gene expression as a novel genomic biomarker in complicated sarcoidosis.
Sarcoidosis, a systemic granulomatous syndrome invariably affecting the lung, typically spontaneously remits but in ~20% of cases progresses with severe lung dysfunction or cardiac and neurologic involvement (complicated sarcoidosis). Unfortunately, current biomarkers fail to distinguish patients with remitting (uncomplicated) sarcoidosis from other fibrotic lung disorders, and fail to identify individuals at risk for complicated sarcoidosis. We utilized genome-wide peripheral blood gene expression analysis to identify a 20-gene sarcoidosis biomarker signature distinguishing sarcoidosis (n = 39) from healthy controls (n = 35, 86% classification accuracy) and which served as a molecular signature for complicated sarcoidosis (n = 17). As aberrancies in T cell receptor (TCR) signaling, JAK-STAT (JS) signaling, and cytokine-cytokine receptor (CCR) signaling are implicated in sarcoidosis pathogenesis, a 31-gene signature comprised of T cell signaling pathway genes associated with sarcoidosis (TCR/JS/CCR) was compared to the unbiased 20-gene biomarker signature but proved inferior in prediction accuracy in distinguishing complicated from uncomplicated sarcoidosis. Additional validation strategies included significant association of single nucleotide polymorphisms (SNPs) in signature genes with sarcoidosis susceptibility and severity (unbiased signature genes - CX3CR1, FKBP1A, NOG, RBM12B, SENS3, TSHZ2; T cell/JAK-STAT pathway genes such as AKT3, CBLB, DLG1, IFNG, IL2RA, IL7R, ITK, JUN, MALT1, NFATC2, PLCG1, SPRED1). In summary, this validated peripheral blood molecular gene signature appears to be a valuable biomarker in identifying cases with sarcoidoisis and predicting risk for complicated sarcoidosis.


Influence of pH on Pb accumulation in the blue mussel, Mytilus edulis.
Changes in seawater pH can alter the chemical speciation of waterborne chemical elements, affecting their bioavailability and, consequently, their bioaccumulation in marine organisms. Here, controlled environmental conditions and a 210Pb radiotracer were used to assess the effect of five distinct pH conditions (pHT ranging from 7.16 to 7.94) on the short-term (9 days) accumulation of Pb in the blue mussel, Mytilus edulis. After 9 days of exposure, higher levels of Pb were observed in the soft tissues of mussels maintained in the lower pH conditions, while Pb levels accumulated by mussel shells showed no difference across pH conditions. These results suggest that pH decreases such as those predicted by ocean acidification scenarios could enhance Pb contamination in marine organisms, with potential subsequent contamination and effect risks for human consumers.


Translocation portals for the substrates and products of a viral transcription complex: the bluetongue virus core.
The bluetongue virus core is a molecular machine that simultaneously and repeatedly transcribes mRNA from 10 segments of viral double-stranded RNA, packaged in a liquid crystalline array. To determine how the logistical problems of transcription within a sealed shell are solved, core crystals were soaked with various ligands and analysed by X-ray crystallography. Mg(2+) ions produce a slight expansion of the capsid around the 5-fold axes. Oligonucleotide soaks demonstrate that the 5-fold pore, opened up by this expansion, is the exit site for mRNA, whilst nucleotide soaks pinpoint a separate binding site that appears to be a selective channel for the entry and exit of substrates and by-products. Finally, nucleotides also bind to the outer core layer, providing a substrate sink.


The gas vesicle gene cluster from Microcystis aeruginosa and DNA rearrangements that lead to loss of cell buoyancy.
Microcystis aeruginosa is a planktonic unicellular cyanobacterium often responsible for seasonal mass occurrences at the surface of freshwater environments. An abundant production of intracellular structures, the gas vesicles, provides cells with buoyancy. A 8.7-kb gene cluster that comprises twelve genes involved in gas vesicle synthesis was identified. Ten of these are organized in two operons, gvpA(I)A(II)A(III)CNJX and gvpKFG, and two, gvpV and gvpW, are individually expressed. In an attempt to elucidate the basis for the frequent occurrence of nonbuoyant mutants in laboratory cultures, four gas vesicle-deficient mutants from two strains of M. aeruginosa, PCC 7806 and PCC 9354, were isolated and characterized. Their molecular analysis unveiled DNA rearrangements due to four different insertion elements that interrupted gvpN, gvpV, or gvpW or led to the deletion of the gvpA(I)-A(III) region. While gvpA, encoding the major gas vesicle structural protein, was expressed in the gvpN, gvpV, and gvpW mutants, immunodetection revealed no corresponding GvpA protein. Moreover, the absence of a gas vesicle structure was confirmed by electron microscopy. This study brings out clues concerning the process driving loss of buoyancy in M. aeruginosa and reveals the requirement for gas vesicle synthesis of two newly described genes, gvpV and gvpW.


Interactions between markers can be caused by the dominance effect of quantitative trait loci.
F(2) populations are commonly used in genetic studies of animals and plants. For simplicity, most quantitative trait locus or loci (QTL) mapping methods have been developed on the basis of populations having two distinct genotypes at each polymorphic marker or gene locus. In this study, we demonstrate that dominance can cause the interactions between markers and propose an inclusive linear model that includes marker variables and marker interactions so as to completely control both additive and dominance effects of QTL. The proposed linear model is the theoretical basis for inclusive composite-interval QTL mapping (ICIM) for F(2) populations, which consists of two steps: first, the best regression model is selected by stepwise regression, which approximately identifies markers and marker interactions explaining both additive and dominance variations; second, the interval mapping approach is applied to the phenotypic values adjusted by the regression model selected in the first step. Due to the limited mapping population size, the large number of variables, and multicollinearity between variables, coefficients in the inclusive linear model cannot be accurately determined in the first step. Interval mapping is necessary in the second step to fine tune the QTL to their true positions. The efficiency of including marker interactions in mapping additive and dominance QTL was demonstrated by extensive simulations using three QTL distribution models with two population sizes and an actual rice F(2) population.


Chagas' disease diagnosis: comparative analysis of parasitologic, molecular, and serologic methods.
During the course of chronic chagasic infection, low parasitemia levels prevent parasite detection by current techniques such as hemoculture and xenodiagnosis. Since serologic tests have sensitivity but lack specificity, molecular assays based on the polymerase chain reaction (PCR) have been proposed as alternative tools for parasite detection in individuals with chronic Chagas' disease. A variable degree of PCR efficiency has been reported in the literature and illustrates the need for further evaluation of large numbers of chagasic patients. In this study, we compared an optimized PCR technique with hemoculture and complement-mediated lysis (CoML) in 113 individuals from or living in endemic areas of Brazil who had conventional serologic results that were either positive, negative, or inconclusive. The PCR amplification yielded positive results in 83.5% (66 of 79) of individuals with positive serology, 47.6% (10 of 21) with negative serology, and 46.2% (6 of 13) with inconclusive serology. Of 10 patients with negative serology and positive PCR result, eight (80%) had positive CoML, indicating that they could have been chagasic but were not mounting immune responses. The PCR results were also positive for all individuals who had positive hemoculture, for 37 individuals with negative hemoculture and positive serology, and for two of six individuals with inconclusive serology and negative hemoculture. Thirteen individuals living in nonendemic areas who had negative serology were used as a negative control group: 100% had negative PCR results. Our results show that the optimized PCR protocol used here was very sensitive in detecting the presence of Trypanosoma cruzi in chronic chagasic patients. The PCR and CoML results were well correlated in all of the groups studied, which suggests that our PCR protocol may be effective in the evaluation of cure in patients who receive anti-parasite treatment.


Prospective Outcomes of Cryopreserved Osteochondral Allograft for Patellofemoral Cartilage Defects at Minimum 2-Year Follow-up.
To analyze the clinical outcomes, knee function, and activity level of patients after treatment of full-thickness cartilage defects involving the patellofemoral compartment of the knee with cryopreserved osteochondral allograft. Nineteen patients with cartilage defects involving the patellofemoral compartment were treated. The average age was 31 years (range 15-45 years), including 12 females and 7 males. Patients were prospectively followed using validated clinical outcome measures including Veterans RAND 12-item Health Survey (VR-12), International Knee Documentation Committee (IKDC), Knee Injury and Osteoarthritis Outcome Score (KOOS), and the Tegner activity scale. Graft incorporation was evaluated by magnetic resonance imaging (MRI) or second-look arthroscopy. The cartilage defects included the patella (n = 16) and the femoral trochlea (n = 3). Mean VR-12 scores increased from 31.6 to 46.3 (P < 0.01), mean IKDC increased from 40.0 to 69.7 (P < 0.01), mean KOOS increased from 53.9 to 80.2 (P < 0.01), and mean Tegner scores increased from 3.0 to 4.9 (P < 0.01), at average follow-up of 41.9 months (range 24-62 months). Of the 3 patients who underwent second-look arthroscopy, all demonstrated a well-incorporated graft. Mean MOCART score for the 6 patients with follow-up MRI was 62.5 (range 25-85). The reoperation rate was 21.1% and 2 patients (12.5%) experienced progressive patellofemoral osteoarthritis requiring conversion to patellofemoral arthroplasty. Patients with unipolar cartilage defects involving the patellofemoral compartment of the knee can have positive outcomes at minimum 2-year follow-up after surgical treatment with a cryopreserved osteochondral allograft when concomitant pathology is also addressed, but the reoperation rate is high and bipolar cartilage lesions may increase the failure rate.


Reduced-intensity conditioning for hematopoietic cell transplant for HLH and primary immune deficiencies.
Allogeneic hematopoietic cell transplantation (HCT) with myeloablative conditioning for disorders associated with excessive inflammation such as hemophagocytic lymphohistiocytosis (HLH) is associated with early mortality. A multicenter prospective phase 2 trial of reduced-intensity conditioning with melphalan, fludarabine, and intermediate-timing alemtuzumab was conducted for HLA matched or single HLA locus mismatched related or unrelated donor HCT in a largely pediatric cohort. Graft-versus-host disease (GVHD) prophylaxis was cyclosporine with methylprednisolone. The primary end point was 1-year overall survival (OS). Thirty-four patients with HLH and 12 with other primary immune deficiencies were transplanted. With a median follow-up of 20 months, the 1-year OS for transplanted patients was 80.4% (90% confidence interval [CI], 68.6%-88.2%). Five additional deaths by 16 months yielded an 18-month OS probability of 66.7% (90% CI, 52.9%-77.3%). Two patients experienced primary graft failure, and 18 patients either experienced a secondary graft failure or required a second intervention (mostly donor lymphocyte infusion [DLI]). At 1 year, the proportion of patients alive with sustained engraftment without DLI or second HCT was 39.1% (95% CI, 25.2%-54.6%), and that of being alive and engrafted (with or without DLI) was 60.9% (95% CI, 45.4 %-74.9%). The day 100 incidence of grade II to IV acute GVHD was 17.4% (95% CI, 8.1%-29.7%), and 1-year incidence of chronic GVHD was 26.7% (95% CI, 14.6%-40.4%). Although the trial demonstrated low early mortality, the majority of surviving patients required DLI or second HCT. These results demonstrate a need for future approaches that maintain low early mortality with improved sustained engraftment. The trial was registered at Clinical Trials.gov (NCT 01998633).


Discovery and cardioprotective effects of the first non-Peptide agonists of the G protein-coupled prokineticin receptor-1.
Prokineticins are angiogenic hormones that activate two G protein-coupled receptors: PKR1 and PKR2. PKR1 has emerged as a critical mediator of cardiovascular homeostasis and cardioprotection. Identification of non-peptide PKR1 agonists that contribute to myocardial repair and collateral vessel growth hold promises for treatment of heart diseases. Through a combination of in silico studies, medicinal chemistry, and pharmacological profiling approaches, we designed, synthesized, and characterized the first PKR1 agonists, demonstrating their cardioprotective activity against myocardial infarction (MI) in mice. Based on high throughput docking protocol, 250,000 compounds were computationally screened for putative PKR1 agonistic activity, using a homology model, and 10 virtual hits were pharmacologically evaluated. One hit internalizes PKR1, increases calcium release and activates ERK and Akt kinases. Among the 30 derivatives of the hit compound, the most potent derivative, IS20, was confirmed for its selectivity and specificity through genetic gain- and loss-of-function of PKR1. Importantly, IS20 prevented cardiac lesion formation and improved cardiac function after MI in mice, promoting proliferation of cardiac progenitor cells and neovasculogenesis. The preclinical investigation of the first PKR1 agonists provides a novel approach to promote cardiac neovasculogenesis after MI.


A multicentre study evaluating the effects of the Swedish ACE programme.
This study investigated the effects of a modified Swedish version of an interactive group education programme, the Active Communication Education (ACE) programme, in five Swedish regions. This study also explored whether the pre- and post-programme outcomes differed with regard to region, age, gender, hearing loss (HL) or the attendance of significant others (SOs). An intervention study with between- and within-group measurements was applied. Seventy-seven individuals with hearing impairments and a mean age of 73.9 years (SD = 9.8) from five different regions in Sweden participated in this study. Statistically significant short- and long-term effects on communication strategy use, activity and participation were observed. The ACE programme was most effective for older individuals, women and participants with more severe HL. Individuals who attended with an SO tended to use better communication strategies. No regional differences were observed. The qualitative results indicated that the programme increased individuals' ability to cope and restored their social identities. The ACE programme is effective, is recommended for implementation in clinical settings and is considered an alternative or additional treatment to hearing aid rehabilitation. Additional studies that include younger individuals and a control group are recommended.


Racial/Ethnic Disparities in Consistent Reporting of Smoking-Related Behaviors.
This study assessed the effect of race/ethnicity on the prevalence of inconsistent reports regarding ever smoking, time since smoking cessation, and age of initiating regular smoking. We used the Tobacco Use Supplement to the Current Population Survey data, which came from a test-retest reliability study, and considered three racial/ethnic subpopulations, Hispanics, Non-Hispanic (NH) Blacks and NH Whites. Initial exploration of highly disagreeing reports of time since smoking cessation and age of onset of regular smoking initiation indicated that the majority of these reports corresponded to NH Whites. However, the proportion of the extremely discrepant reports was very small (less than 0.8%), and these reports were not included in the main analyses. Univariate analyses revealed that for each smoking measure, NH Whites tended to report most consistently when compared to Hispanics and NH Blacks. However, the only statistically significant result was that Hispanics were more likely to report their regular smoking initiation age inconsistently than were NH Whites. Analyses that adjusted for other factors confirmed this finding, i.e., Hispanics were 1.8 times more likely to provide inconsistent reports of their age of onset of regular smoking than were NH Whites. Furthermore, these analyses showed that the impact of race/ethnicity on the prevalence of inconsistent reporting may depend on other factors, e.g., age and employment status. For example, non-employed NH Blacks were 1.9 times more likely to recant ever smoking than were non-employed NH Whites. The lower consistency in reports by Hispanics and NH Blacks underscores the importance of developing new survey design and research strategies for detecting relatively small differences in reporting among the racial/ethnic minorities. Additional efforts to motivate racial/ethnic minorities to participate in national surveys may not only help increase representation of these subpopulations in study samples but also help improve overall data quality.


Vaginal Microbiota and Cytokine Levels Predict Preterm Delivery in Asian Women.
Preterm birth (PTB) is the most common cause of neonatal morbidity and mortality worldwide. Approximately half of PTBs is linked with microbial etiologies, including pathologic changes to the vaginal microbiota, which vary according to ethnicity. Globally more than 50% of PTBs occur in Asia, but studies of the vaginal microbiome and its association with pregnancy outcomes in Asian women are lacking. This study aimed to longitudinally analyzed the vaginal microbiome and cytokine environment of 18 Karen and Burman pregnant women who delivered preterm and 36 matched controls delivering at full term. Using 16S ribosomal RNA gene sequencing we identified a predictive vaginal microbiota signature for PTB that was detectable as early as the first trimester of pregnancy, characterized by higher levels of Prevotella buccalis, and lower levels of Lactobacillus crispatus and Finegoldia, accompanied by decreased levels of cytokines including IFNγ, IL-4, and TNFα. Differences in the vaginal microbial diversity and local vaginal immune environment were associated with greater risk of preterm birth. Our findings highlight new opportunities to predict PTB in Asian women in low-resource settings who are at highest risk of adverse outcomes from unexpected PTB, as well as in Burman/Karen ethnic minority groups in high-resource regions.


Paroxysmal dyskinesias.
Paroxysmal dyskinesias are a rare group of movement disorders affecting both adults and children. Based on the events that precipitate the abnormal movements, they are subdivided into paroxysmal kinesigenic dyskinesia (PKD), precipitated by sudden voluntary movements; paroxysmal nonkinesigenic dyskinesia (PNKD), which occurs at rest; paroxysmal exertion-induced dyskinesia (PED), occurring after prolonged exercise; and paroxysmal hypnogenic dyskinesia (PHD), which occurs in sleep. Paroxysmal dyskinesias can be sporadic, familial (autosomal dominant inheritance), or secondary to other disorders. Recent genetic discoveries may aid us in elucidating the pathophysiology of these disorders. PKD has been linked to the pericentromeric region of chromosome 16, PNKD is associated with mutations in the myofibrillogenesis regulator 1 (MR-1) gene on the long arm of chromosome 2 (2q32-36 locus), and PED is associated with mutations in the glucose transporter gene, GLUT1, responsible for glucose transport across the blood-brain barrier. Lifestyle modification to avoid precipitating factors is important in the management of paroxysmal dyskinesias. Medical therapies have not been examined in controlled trials. Nevertheless, anticonvulsants have been found to be extremely effective in treating PKD and are sometimes useful in other types, suggesting that these disorders may indeed represent forms of channelopathies. Drugs such as acetazolamide, anticholinergics, levodopa, and tetrabenazine have been inconsistently successful. In rare cases with medically refractory symptoms, deep brain stimulation has also been employed. Development of successful treatments for the different paroxysmal dyskinesias rests on elucidating the pathophysiology and targeting therapy to treat the underlying perturbation.


Interaction of adrenocortical activity and autonomic arousal on children's externalizing and internalizing behavior problems.
The psychobiology of stress involves two major components, the hypothalamic-pituitary-adrenal (HPA) axis and the autonomic nervous system (ANS). Research has revealed the association between behavior problems and the psychobiology of stress, yet findings are inconsistent and few studies have addressed the moderate correlations between behavior problems. This study examines the individual and interactive effects of HPA and ANS on child behavior problems while taking into account the comorbidity of externalizing and internalizing problems. Four saliva samples were collected from each participant in a community sample (N = 429; aged 11-12 years; 50.49 % male), which were assayed for cortisol (HPA) and alpha-amylase, sAA (ANS). Children's behavior problems were assessed using parent-report and self-report versions of the Child Behavior Checklist. Latent variables were constructed to represent trait-like individual differences in cortisol and sAA. Low levels of HPA axis activity were associated with higher levels of both externalizing and internalizing problems, but only among children with low ANS arousal. The association between externalizing and internalizing problems diminished to non-significant after taking into account the influence of HPA axis activity and ANS arousal, which suggests that the psychobiology of stress explains a fair proportion of comorbidity of behavior problems. The findings support that interaction between HPA axis and ANS functioning has potential to clarify prior mixed findings and advance our understanding of the child behavior problems.


Oligomers, fact or artefact? SDS-PAGE induces dimerization of β-amyloid in human brain samples.
The formation of low-order oligomers of β-amyloid (Aβ) within the brain is widely believed to be a central component of Alzheimer's disease (AD) pathogenesis. However, despite advances in high-throughput and high-resolution techniques such as xMAP and mass spectrometry (MS), investigations into these oligomeric species have remained reliant on low-resolution Western blots and enzyme-linked immunosorbent assays. The current investigation compared Aβ profiles within human cortical tissue using sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis (PAGE), xMAP and surface enhanced laser desorption/ionization time-of-flight MS and found that whilst there was significant correlation across the techniques regarding levels of monomeric Aβ, only SDS-PAGE was capable of detecting dimeric isoforms of Aβ. The addition of synthetic di-tyrosine cross-linked Aβ(1-40)Met(35)(O) to the AD tissue demonstrated that the MS methodology was capable of observing dimeric Aβ at femto-molar concentrations, with no noticeable effect on monomeric Aβ levels. Focus turned to the association between SDS-PAGE and levels of observable dimeric Aβ within the AD brain tissue. These investigations revealed that increased levels of dimeric Aβ were observed with increasing concentrations of SDS in the sample buffer. This finding was subsequently confirmed using synthetic Aβ(1-42) and suggests that SDS was inducing the formation of dimeric Aβ. The findings that SDS promotes Aβ dimerization have significant implications for the putative role of low-order oligomers in AD pathogenesis and draw into question the utility of oligomeric Aβ as a therapeutic target.


Prospective evidence for independent nitrogen and phosphorus limitation of grasshopper (Chorthippus curtipennis) growth in a tallgrass prairie.
Insect herbivores play a pivotal role in regulating plant production and community composition, and their role in terrestrial ecosystems is partly determined by their feeding behavior and performance among plants of differing nutritional quality. Historically, nitrogen (N) has been considered the primary limiting nutrient of herbivorous insects, but N is only one of many potential nutrients important to insect performance. Of these nutrients, phosphorus (P) is perhaps the most important because somatic growth depends upon P-rich ribosomal RNA. Yet relatively few studies have assessed the strength of P-limitation for terrestrial insects and even fewer have simultaneously manipulated both N and P to assess the relative strengths of N- and P-limitation. Here, we tested for potential N and P limitation, as well as N:P co-limitation, on Chorthippis curtipennis (Orthoptera, Acrididae), an abundant member of arthropod communities of central US prairies. Our results demonstrate weak evidence for both N and P limitation of C. curtipennis growth rates in laboratory feeding assays. Importantly, P-limitation was just as strong as N-limitation, but we found no evidence for NP co-limitation in our study. Furthermore, nutrient limitation was not apparent in field studies, suggesting that insect growth rates may be predominately controlled by other factors, including temperature and predation. Our results suggest that P should be jointly considered, along with N, as a primary determinant of herbivore feeding behavior under both current and future climate conditions.


Indications and Effects of Plasma Transfusions in Critically Ill Children.
Plasma transfusions are frequently prescribed for critically ill children, although their indications lack a strong evidence base. Plasma transfusions are largely driven by physician conceptions of need, and these are poorly documented in pediatric intensive care patients. To identify patient characteristics and to characterize indications leading to plasma transfusions in critically ill children, and to assess the effect of plasma transfusions on coagulation tests. Point-prevalence study in 101 pediatric intensive care units in 21 countries, on 6 predefined weeks. All critically ill children admitted to a participating unit were included if they received at least one plasma transfusion. During the 6 study weeks, 13,192 children were eligible. Among these, 443 (3.4%) received at least one plasma transfusion and were included. The primary indications for plasma transfusion were critical bleeding in 22.3%, minor bleeding in 21.2%, planned surgery or procedure in 11.7%, and high risk of postoperative bleeding in 10.6%. No bleeding or planned procedures were reported in 34.1%. Before plasma transfusion, the median international normalized ratio (INR) and activated partial thromboplastin time (aPTT) values were 1.5 and 48, respectively. After plasma transfusion, the median INR and aPTT changes were -0.2 and -5, respectively. Plasma transfusion significantly improved INR only in patients with a baseline INR greater than 2.5. One-third of transfused patients were not bleeding and had no planned procedure. In addition, in most patients, coagulation tests are not sensitive to increases in coagulation factors resulting from plasma transfusion. Studies assessing appropriate plasma transfusion strategies are urgently needed.


Use of a field portable X-Ray fluorescence analyzer to determine the concentration of lead and other metals in soil samples.
Field portable methods are often needed in risk characterization, assessment and management to rapidly determine metal concentrations in environmental samples. Examples are for determining: "hot spots" of soil contamination, whether dust wipe lead levels meet housing occupancy standards, and worker respiratory protection levels. For over 30 years portable X-Ray Fluorescence (XRF) analyzers have been available for the in situ, non-destructive, measurement of lead in paint. Recent advances made possible their use for analysis of airborne dust filter samples, soil, and dust wipes. Research at the University of Cincinnati with the NITON 700 Series XRF instrument (40 millicurie Cadmium 109 source, L X-Rays) demonstrated its proficiency on air sample filters (NIOSH Method No. 7702, "Lead by Field Portable XRF; limit of detection 6 microg per sample; working range 17-1,500 microg/m3 air). Research with lead dust wipe samples from housing has also shown promising results. This XRF instrument was used in 1997 in Poland on copper smelter area soil samples with the cooperation of the Wroclaw Medical Academy and the Foundation for the Children from the Copper Basin (Legnica). Geometric mean soil lead concentrations were 200 ppm with the portable XRF, 201 ppm with laboratory-based XRF (Kevex) and 190 ppm using atomic absorption (AA). Correlations of field portable XRF and AA results were excellent for samples sieved to less than 125 micrometers with R-squared values of 0.997, 0.957, and 0.976 for lead, copper and zinc respectively. Similarly, correlations were excellent for soil sieved to less than 250 micrometers, where R-squared values were 0. 924, 0.973, and 0.937 for lead, copper and zinc, respectively. The field portable XRF instrument appears to be useful for the determination of soil pollution by these metals in industrial regions.


Fine needle aspiration biopsy with adjunct immunohistochemistry in intraocular tumor management.
To assess the effectiveness of fine needle aspiration biopsy (FNAB), with and without immunohistochemistry (IHC), in the management of solid intraocular tumors. Thirty-three consecutive adults undergoing FNAB of suspected intraocular tumors were studied. Clinical, cytologic and histologic diagnoses were correlated. The positive predictive value, sensitivity and specificity of FNAB for detecting malignancy, the effect of lHC on the final cytologic diagnosis and the number of patients in whom clinical management was altered as a result of cytologic evaluation were determined. The positive predictive value was 96% with and 93% without adjunct IHC. The sensitivity and specificity of FNAB for detecting malignancy were 96% and 83%, respectively, with IHC. Without IHC, the sensitivity was unaltered, but the specificity was 67%. IHC confirmed the morphologic diagnosis in 75% of cases, made a diagnosis in 12.5% and changed a malignant diagnosis from carcinoma to melanoma in 6% of cases. The planned management was changed by the FNAB findings in 24% of patients. In 3 patients (9%), IHC was essential for diagnosis and management. No patients exhibited local tumor dissemination or recurrence associated with the biopsy. FNAB is a safe, sensitive and specific method of establishing a tissue diagnosis in a subset of patients with solid intraocular tumors. The routine use of immunohistochemical stain ing increases the diagnostic utility of the technique and may change clinical management.


Biases in attentional orientation and magnitude estimation explain crossover: neglect is a disorder of both.
Crossover refers to a pattern of performance on the line bisection test in which short lines are bisected on the side opposite the true center of long lines. Although most patients with spatial neglect demonstrate crossover, contemporary theories of neglect cannot explain it. In contrast, we show that blending the psychophysical construct of magnitude estimation with neglect theory not only explains crossover, but also addresses a quantitative feature of neglect that is independent of spatial deficits. We report a prospective validation study of the orientation/estimation hypothesis of crossover. Forty subjects (17 patients with and without neglect following unilateral brain injury and 23 normal controls) completed four experiments that examined crossover using line bisection, line bisection with cueing, and reproducing line lengths from both memory and a standard. Replicating earlier findings, all except one subject group exhibited crossover on the standard line bisection test, all groups showed a spontaneous preference to orient attention to one end of the lines, and all groups overestimated the length of short lines and underestimated long lines. Biases in attentional orientation and magnitude estimation are exaggerated in patients with neglect. The truly novel finding of this study occurred when, after removing the line from the bisection task, the direction of crossover was completely reversed in all subject groups depending on where attention was oriented. These findings are consistent with our hypothesis of crossover: (1) crossover is a normal component of performance on line bisection; (2) crossover results from the interplay of biases in attentional orientation and magnitude estimation; and (3) attentional orientation predicts the direction of crossover, whereas a disorder of magnitude estimation, not previously emphasized in neglect, accounts for the quantitative changes in length estimation that make crossover more obvious in neglect subjects. Paradoxically, we observed that the traditional line bisection test is suboptimal for exploring crossover because lines elicit spontaneous orientation responses from subjects that confound experimental manipulations of attention. We conclude that attentional orientation and magnitude estimation are necessary and sufficient to explain crossover and that bias in magnitude estimation is a core component of neglect.


Recent development and new ideas in the field of dispersive multilayer optics.
A dispersive-mirror-based laser permits a dramatic simplification of high-power femtosecond and attosecond systems and affords promise for their further development toward shorter pulse durations, higher peak powers, and higher average powers with user-friendly systems. The result of the continuous development of dispersive mirrors permits pulse compression down to almost single cycle pulses of 3 fs duration. These design approaches together with the existing modern deposition technology pave the way for the manufacture of dielectric multilayer coatings able to compress pulses of tens of picoseconds duration down to a few femtoseconds.


Photodynamic therapy for prostate cancer--a review of current status and future promise.
Debate is ongoing about the treatment of organ-confined prostate cancer, particularly in men who have low-risk disease detected by PSA screening. A balance is needed between the harms and benefits of treatment. New techniques are being developed that aim to offer similar treatment effects to current radical therapies, while reducing the associated harmful effects of these treatments. In this Review, we explore the potential of one such technique, photodynamic therapy (PDT), for the treatment of organ-confined prostate cancer. PDT uses a photosensitizing drug that is activated in the prostate by low-power laser light, delivered using optical fibers. The fibers are placed within needles in the prostate, guided by transrectal ultrasound and a perineal template. Following the activation of the photosensitizer by light, and the formation of reactive oxygen species, necrosis occurs at the site of interaction between the photosensitizer, light and oxygen. Clinical studies are underway to investigate the use of PDT for primary and salvage treatment of organ-confined prostate cancer. We review these studies, the potential strategies for enhanced photodynamic effects, and the current limitations of PDT for prostate cancer.


Influence of age and body mass on the response of adolescent male pigs to anaesthesia.
Age and body mass affect a human's response to drugs, including anaesthetics. In pigs, such effects, if they occur, are likely to be pronounced: commercial breeds have been selected for rapid growth, meaning rapid body composition and mass change with age. Thirty-six male pigs were anaesthetised for CT scanning on three occasions (S1-3) when aged 105, 137 and 166 days and when mean (±sd) masses were 57.2±4.4, 88.4±6.2 and 114.7±7.6 kg, respectively. Medetomidine (5 μg/kg), azaperone (1 mg/kg), ketamine (5 mg/kg) and midazolam (0.25 mg/kg) were combined and injected intramuscularly. The times when pigs became recumbent (R1) and remained so (RP) were recorded. If venous cannulation was not possible five minutes after recumbency, 2-3 per cent isoflurane in a 1:2 O2/N2O mixture was delivered by mask until cannulation was possible and then discontinued. If anaesthetic depth was inadequate for CT scanning, a full dose (midazolam 0.25 mg/kg, ketamine 2 mg/kg) or half dose of induction agents was administered intravenously. During recovery from anaesthesia, the times at first movement (M1), first standing attempt (S1) and successful sustained standing (SP) were recorded. The relationship between mass and time (minutes) from injection to each end point was assessed using regression analysis and linear mixed-effect models (LMEM); LMEM were used to assess isoflurane and intravenous anaesthetic effects. Analysis using LMEM showed no significant relationships between mass and the times from injection to the five end points. Isoflurane reduced the time to M1, S1 and SP (P<0.037); intravenous agents had no effect on S1 or SP (P>0.585) but increased the time from injection to M1 (P<0.001). In conclusion, age and mass do not influence the response of commercially bred pigs to the intravenous anaesthetic combination described.


The effect of respiratory and cardiac motion in liver diffusion tensor imaging (DTI).
To evaluate the effect of respiratory and cardiac motion on diffusion tensor imaging (DTI) metrics in healthy human liver. Fifteen healthy subjects, participating in either part of this study, were scanned using a 1.5-T magnetic resonance imaging (MRI) device. Coronal liver DTI (6 diffusion-encoding directions; b, 300 mm/s) during breath holding was compared to free breathing. Cardiac motion effects were evaluated by comparing breath-held DTI scans acquired during both diastole and systole. Free breathing resulted in a significantly increased mean diffusivity (P < 0.05), λ1 (P < 0.01), λ2 (P < 0.05), and λ3 (P < 0.01) compared to breath holding. During systole significant increases in fractional anisotropy (P < 0.05), mean diffusivity (P < 0.05), and λ1 (P < 0.05), compared to systole, were found in the left lobe. The right lobe, which is less affected by cardiac motion, showed no significant change in DTI metrics over the cardiac cycle. Respiratory and cardiac motion tends to increase liver DTI metrics.


DNA replication at the single-molecule level.
A cell can be thought of as a highly sophisticated micro factory: in a pool of billions of molecules - metabolites, structural proteins, enzymes, oligonucleotides - multi-subunit complexes assemble to perform a large number of basic cellular tasks, such as DNA replication, RNA/protein synthesis or intracellular transport. By purifying single components and using them to reconstitute molecular processes in a test tube, researchers have gathered crucial knowledge about mechanistic, dynamic and structural properties of biochemical pathways. However, to sort this information into an accurate cellular road map, we need to understand reactions in their relevant context within the cellular hierarchy, which is at the individual molecule level within a crowded, cellular environment. Reactions occur in a stochastic fashion, have short-lived and not necessarily well-defined intermediates, and dynamically form functional entities. With the use of single-molecule techniques these steps can be followed and detailed kinetic information that otherwise would be hidden in ensemble averaging can be obtained. One of the first complex cellular tasks that have been studied at the single-molecule level is the replication of DNA. The replisome, the multi-protein machinery responsible for copying DNA, is built from a large number of proteins that function together in an intricate and efficient fashion allowing the complex to tolerate DNA damage, roadblocks or fluctuations in subunit concentration. In this review, we summarize advances in single-molecule studies, both in vitro and in vivo, that have contributed to our current knowledge of the mechanistic principles underlying DNA replication.


An anti-platelet-endothelial cell adhesion molecule-1 antibody inhibits leukocyte extravasation from mesenteric microvessels in vivo by blocking the passage through the basement membrane.
Platelet-endothelial cell adhesion molecule-1 (PECAM-1, CD31) plays an active role in the process of leukocyte migration through cultured endothelial cells in vitro and anti-PECAM-1 antibodies (Abs) inhibit accumulation of leukocytes into sites of inflammation in vivo. Despite the latter, it is still not clear at which stage of leukocyte emigration in vivo PECAM-1 is involved. To address this point directly, we studied the effect of an anti-PECAM-1 Ab, recognizing rat PECAM-1, on leukocyte responses within rat mesenteric microvessels using intravital microscopy. In mesenteric preparations activated by interleukin (IL)-1 beta, the anti-PECAM-1 Ab had no significant effect on the rolling or adhesion of leukocytes, but inhibited their migration into the surrounding extravascular tissue in a dose-dependent manner. Although in some vessel segments these leukocytes had come to a halt within the vascular lumen, often the leukocytes appeared to be trapped within the vessel wall. Analysis of these sections by electron microscopy revealed that the leukocytes had passed through endothelial cell junctions but not the basement membrane. In contrast to the effect of the Ab in mesenteric preparations treated with IL-1 beta, leukocyte extravasation induced by topical or intraperitoneal administration of the chemotactic peptide formyl-methionyl-leucyl-phenylalanine was not inhibited by the anti-PECAM-1 Ab. These results directly demonstrate a role for PECAM-1 in leukocyte extravasation in vivo and indicate that this involvement is selective for leukocyte extravasation elicited by certain inflammatory mediators. Further, our findings provide the first in vivo indication that PECAM-1 may have an important role in triggering the passage of leukocytes through the perivascular basement membrane.


Nurses' use of online clinical evidence.
The 'information explosion' presents a significant challenge to the nursing profession. One way to assist access to information is to provide electronic information resources at the point of care. A large-scale survey of nurses working in the public health system in New South Wales, Australia, was undertaken as part of a 2-year evaluation of the Clinical Information Access Program (CIAP). This is a website which provides 24 hour, online access to a range of evidence sources at the point of care. This paper reports a study to investigate factors influencing nurses' use of online evidence available at the point of care and to examine differences between nurses in different roles. Nurses from 65 randomly selected hospitals were surveyed. A convenience sample representing 25% of nursing staff at these hospitals was sought. Eighty-four per cent of the survey quota was met and 3128 questionnaires were completed and returned. The results showed that 58% of nurses had heard of the CIAP and, of those, 70% had used the website's resources. Senior nurses had greater awareness and use than others. The most frequent reasons for use were to fill a knowledge gap and for personal education. Lack of training was the most frequently cited reason for not using the website. Use of online evidence, particularly amongst senior staff, was higher than reported in previous studies. Use of online evidence was associated with nursing role, and with managerial and organizational support. Diffusion of innovation theory can help to explain some of the patterns observed. The use and impact of online evidence should be interpreted in the context of nursing practice and culture.


Consequences of toxic secondary compounds in nectar for mutualist bees and antagonist butterflies.
Attraction of mutualists and defense against antagonists are critical challenges for most organisms and can be especially acute for plants with pollinating and non-pollinating flower visitors. Secondary compounds in flowers have been hypothesized to adaptively mediate attraction of mutualists and defense against antagonists, but this hypothesis has rarely been tested. The tissues of milkweeds (Asclepias spp.) contain toxic cardenolides that have long been studied as chemical defenses against herbivores. Milkweed nectar also contains cardenolides, and we have examined the impact of manipulating cardenolides in nectar on the foraging choices of two flower visitors: generalist bumble bees, Bombus impatiens, which are mutualistic pollinators, and specialist monarch butterflies, Danaus plexippus, which are herbivores as larvae and ineffective pollinators as adults. Although individual bumble bees in single foraging bouts showed no avoidance of cardenolides at the highest natural concentrations reported for milkweeds, a pattern of deterrence did arise when entire colonies were allowed to forage for several days. Monarch butterflies were not deterred by the presence of cardenolides in nectar when foraging from flowers, but laid fewer eggs on plants paired with cardenolide-laced flowers compared to controls. Thus, although deterrence of bumble bees by cardenolides may only occur after extensive foraging, a primary effect of nectar cardenolides appears to be reduction of monarch butterfly oviposition.


Delivery of transforming growth factor-beta2-perturbing antibody in a collagen vehicle inhibits cranial suture fusion in calvarial organ culture.
To determine whether antibody perturbation of Tgf-beta, delivered in a collagen gel, could inhibit cranial suture fusion. Attachment and proliferation of osteoblasts cultured on a collagen gel with or without anti-Tgf-beta2 antibody were determined by AlamarBlue dye assay and cell morphology by toluidine-blue staining. In rat calvarial organ culture, collagen gel with and without anti-Tgf-beta2 antibody was injected subperiosteally over the posterior frontal suture of postnatal day 15 rat calvariae. A quantitative analysis of suture fusion was used to measure suture bridging in histological serial sections at various time points. Attachment and proliferation for cells cultured on collagen gel with anti-Tgf-beta2 antibody were similar to collagen gel controls. Although proliferation was lower than on tissue culture plastic, cells treated with anti-Tgf-beta2 antibody maintained an osteoblastic morphology. After 7, 10, and 15 days in organ culture, anti-Tgf-beta2 antibody treatment caused a reduction in the percent bridging of posterior frontal sutures, compared with controls. Sutures exposed to anti-Tgf-beta2 antibody and fibroblast growth factor-2 concurrently did not show an inhibition of bony bridging. These results support previous reports suggesting a role for Tgf-beta2 in cranial suture fusion. In cell culture the collagen gel, both with and without anti-Tgf-beta2 antibody, promoted similar osteoblast attachment, proliferation, and osteoblastic morphology. In organ culture anti-Tgf-beta2 antibody was delivered in a bioactive state via a collagen gel to inhibit cranial suture fusion. Also, the results suggest that the inductive effect of fibroblast growth factor-2 is not dependent on Tgf-beta2 activity. Together, these results provide further support for the role of Tgf-beta2 in cranial suture fusion.


Multisensor system for isotemporal measurements to assess indoor climatic conditions in poultry farms.
The rearing of poultry for meat production (broilers) is an agricultural food industry with high relevance to the economy and development of some countries. Periodic episodes of extreme climatic conditions during the summer season can cause high mortality among birds, resulting in economic losses. In this context, ventilation systems within poultry houses play a critical role to ensure appropriate indoor climatic conditions. The objective of this study was to develop a multisensor system to evaluate the design of the ventilation system in broiler houses. A measurement system equipped with three types of sensors: air velocity, temperature and differential pressure was designed and built. The system consisted in a laptop, a data acquisition card, a multiplexor module and a set of 24 air temperature, 24 air velocity and two differential pressure sensors. The system was able to acquire up to a maximum of 128 signals simultaneously at 5 second intervals. The multisensor system was calibrated under laboratory conditions and it was then tested in field tests. Field tests were conducted in a commercial broiler farm under four different pressure and ventilation scenarios in two sections within the building. The calibration curves obtained under laboratory conditions showed similar regression coefficients among temperature, air velocity and pressure sensors and a high goodness fit (R(2) = 0.99) with the reference. Under field test conditions, the multisensor system showed a high number of input signals from different locations with minimum internal delay in acquiring signals. The variation among air velocity sensors was not significant. The developed multisensor system was able to integrate calibrated sensors of temperature, air velocity and differential pressure and operated successfully under different conditions in a mechanically-ventilated broiler farm. This system can be used to obtain quasi-instantaneous fields of the air velocity and temperature, as well as differential pressure maps to assess the design and functioning of ventilation system and as a verification and validation (V&V) system of Computational Fluid Dynamics (CFD) simulations in poultry farms.


Sentinel node status predicts survival in thick melanomas: the Oxford perspective.
To determine the prognostic value of SLNB in patients with thick melanoma in terms of overall survival (OS) and recurrence-free survival (RFS). 136 patients with primary tumours (Breslow thickness ≥ 4.0 mm) underwent SLNB. OS and RFS were calculated and a multivariate Cox regression model used to determine the important prognostic factors for predicting OS and RFS. Median Breslow thickness was 5.5 mm and 60% were ulcerated. Median follow up was 4 years (95% CI = 4-5) with 54 patients having died at the time of analysis. 5-year OS for SLNB positive patients was 32%, compared to 78% for negative patients. The significant predictors of poorer OS were increasing age (p = 0.03), increasing Breslow thickness (p = 0.03) and SLNB positivity (p < 0.0001). 5 year RFS was significantly worse in the SLNB positive population compared to the negative patients (p < 0.0001); 27% versus 66% respectively. Patients with a thick melanoma and a positive SLNB have a significantly worse RFS and OS compared to those with a negative SLNB. Over three-quarters of patients with a negative SLNB survived five years. These findings have implications for the subpopulations included in adjuvant therapy trials and we advocate SLNB be recommended in patients with thick melanomas.


MR-guided perineural injection of the ganglion impar: technical considerations and feasibility.
Perineural ganglion impar injections are used in the management of pelvic pain syndromes; however, there is no consensus regarding the optimal image guidance. Magnetic resonance imaging (MRI) provides high soft tissue contrast and the potential to directly visualize and target the ganglion. The purpose of this study was to assess the feasibility of MR-guided percutaneous perineural ganglion impar injections. Six MR-guided ganglion impar injections were performed in six human cadavers. Procedures were performed with a clinical 1.5-Tesla MRI system through a far lateral transgluteus approach. Ganglion impar visibility, distance from the sacrococcygeal joint, number of intermittent MRI control steps required to place the needle, target error between the intended and final needle tip location, inadvertent punctures of non-targeted vulnerable structures, injectant distribution, and procedure time were determined. The ganglion impar was seen on MRI in 4/6 (66 %) of cases and located 0.8 mm cephalad to 16.3 mm caudad (average 1.2 mm caudad) to the midpoint of the sacrococcygeal joint. Needle placement required an average of three MRI control steps (range, 2-6). The average target error was 2.2 ± 2.1 mm. In 6/6 cases (100 %), there was appropriate periganglionic distribution and filling of the presacrococcygeal space. No punctures of non-targeted structures occurred. The median procedure time was 20 min (range, 12-29 min). Interventional MRI can visualize and directly target the ganglion impar for accurate needle placement and successful periganglionic injection with the additional benefit of no ionizing radiation exposure to patient and staff. Our results support clinical evaluation.


Prevalence, trends and risk factors of thinness among Greek children and adolescents.
Thinness affects more children and adolescents than obesity. Thus, the aim of the study is to examine the recent estimates of thinness and associated risk factors, and to identify trends in thinness, among Greek schoolchildren. Epidemiological study. Population data from the recent estimates are derived from a school-based health survey polled in 2015 on 336,014 participants aged 4- to 17-years-old. To assess trends of thinness (1996-2015) we included a total of 300,104 children aged 8- to 9-years-old. Physical activity, dietary habits and sedentary activities were assessed through self-completed questionnaires. The gender and age-specific body mass index cut-off points proposed by International Obesity Task Force were used in order to define weight groups. Percent 8.4% of girls and 6.5% of boys were thin (all grades included). The prevalence of thinness decreased with age more in boys (from 13.8% at 4-years-old to 5.1% at 17-years-old, p < 0.001), than in girls (from 10.9% at 4-years-old to 8.7% at 17-years-old, p < 0.001). Sufficient dietary habits (OR: 0.87, 95% CI: 0.77-0.97) and adequate physical activity levels (OR: 0.92, 95% CI: 0.85-0.99) were associated with decreased risk of thinness. Thin schoolchildren performed better in aerobic fitness test than normalweight ones. Between 1996 and 2015, thinness rates decreased from 8.0% to 6.5% in boys (p = 0.046) and from 10.6% to 8.4% in girls (p = 0.036). Our results suggest that thinness is a significant overlooked phenomenon. Although the prevalence of thinness has decreased the last two decades among Greek schoolchildren, actions need to be taken from public policy makers in order to establish and maintain a healthy body weight.


Characterization of the FAD2 Gene Family in Soybean Reveals the Limitations of Gel-Based TILLING in Genes with High Copy Number.
Soybean seed oil typically contains 18-20% oleic acid. Increasing the content of oleic acid is beneficial for health and biodiesel production. Mutations in FAD2-1 genes have been reported to increase seed oleic acid content. A subset of 1,037 mutant families from a mutagenized soybean cultivar (cv.) Forrest population was screened using reverse genetics (TILLING) to identify mutations within FAD2 genes. Although no fad2 mutants were identified using gel-based TILLING, four fad2-1A and one fad2-1B mutants were identified to have high seed oleic acid content using forward genetic screening and subsequent target sequencing. TILLING has been successfully used as a non-transgenic reverse genetic approach to identify mutations in genes controlling important agronomic traits. However, this technique presents limitations in traits such as oil composition due to gene copy number and similarities within the soybean genome. In soybean, FAD2 are present as two copies, FAD2-1 and FAD2-2. Two FAD2-1 members: FAD2-1A and FAD2-1B; and three FAD2-2 members: FAD2-2A, FAD2-2B, and FAD2-2C have been reported. Syntenic, phylogenetic, and in silico analysis revealed two additional members constituting the FAD2 gene family: GmFAD2-2D and GmFAD2-2E, located on chromosomes 09 and 15, respectively. They are presumed to have diverged from other FAD2-2 members localized on chromosomes 19 (GmFAD2-2A and GmFAD2-2B) and 03 (GmFAD2-2C). This work discusses alternative solutions to the limitations of gel-based TILLING in functional genomics due to high copy number and multiple paralogs of the FAD2 gene family in soybean.


Mining basic active structures from a large-scale database.
The Pubchem Database is a large-scale resource for chemical information, containing millions of chemical compound activities derived by high-throughput screening (HTS). The ability to extract characteristic substructures from such enormous amounts of data is steadily growing in importance. Compounds with shared basic active structures (BASs) exhibiting G-protein coupled receptor (GPCR) activity and repeated dose toxicity have been mined from small datasets. However, the mining process employed was not applicable to large datasets owing to a large imbalance between the numbers of active and inactive compounds. In most datasets, one active compound will appear for every 1000 inactive compounds. Most mining techniques work well only when these numbers are similar. This difficulty was overcome by sampling an equal number of active and inactive compounds. The sampling process was repeated to maintain the structural diversity of the inactive compounds. An interactive KNIME workflow that enabled effective sampling and data cleaning processes was created. The application of the cascade model and subsequent structural refinement yielded the BAS candidates. Repeated sampling increased the ratio of active compounds containing these substructures. Three samplings were deemed adequate to identify all of the meaningful BASs. BASs expressing similar structures were grouped to give the final set of BASs. This method was applied to HIV integrase and protease inhibitor activities in the MDL Drug Data Report (MDDR) database and to procaspase-3 activators in the PubChem BioAssay database, yielding 14, 12, and 18 BASs, respectively. The proposed mining scheme successfully extracted meaningful substructures from large datasets of chemical structures. The resulting BASs were deemed reasonable by an experienced medicinal chemist. The mining itself requires about 3 days to extract BASs with a given physiological activity. Thus, the method described herein is an effective way to analyze large HTS databases.


The low-affinity nerve growth factor receptor p75NTR identifies a transient stem cell-like state in oral squamous cell carcinoma cells.
Although several markers have been used for enrichment of cells with stem cell-like properties in oral squamous cell carcinoma (OSCC), isolation of a pure subpopulation is still a challenging task. Normal oral and esophageal keratinocyte stem cells have been previously isolated using the low-affinity nerve growth factor receptor p75NTR. To investigate the potential of p75NTR as a marker for identification and isolation of oral cancer cells with stem cell-like properties. Subpopulations of cells with high or low expression of p75NTR were sorted from OSCC-derived cells and compared for sphere/colony formation, in vivo tumor formation ability, expression of stem cell-related molecules, cell cycle distribution and drug resistance. p75NTR(High) cells exhibited statistically significant higher stem cell properties than p75NTR(Low) cells in all assays performed. Nevertheless, p75NTR(Low) subpopulation did also exhibit some stem cell features, but to a lesser extent. Propagation of p75NTR(Low) cells for several passages in culture showed that the expression of p75NTR could rise spontaneously. This finding was also supported by the similar expression of p75NTR by the xenografts generated by both subpopulations in NOD\SCID IL2Rg(null) mice. p75NTR can be used for isolating a subpopulation enriched for cells with stem cell-like properties in OSCC. De novo generation of p75NTR(High) cells from p75NTR(Low) cells suggests either that there is another subpopulation with stem cell features within the p75NTR(Low) cells, or that the p75NTR(Low) cells can dedifferentiate due to a contextually regulated equilibrium between stem cell-like cells and transit-amplifying neoplastic progenitors.


Effect of the look AHEAD study intervention on medication use and related cost to treat cardiovascular disease risk factors in individuals with type 2 diabetes.
To examine the effect of a lifestyle intervention to produce weight loss and increased physical fitness on use and cost of medications to treat cardiovascular disease (CVD) risk factors in people with type 2 diabetes. Look AHEAD is a multicenter randomized controlled trial of 5,145 overweight or obese individuals with type 2 diabetes, aged 45-76 years. An intensive lifestyle intervention (ILI) involving group and individual meetings to achieve and maintain weight loss through decreased caloric intake and increased physical activity was compared with a diabetes support and education (DSE) condition. Medications prescribed to treat diabetes, hypertension, and hyperlipidemia were compared at baseline and 1 year. Medication costs were conservatively estimated using prices from a national online pharmacy. Participants randomized to an ILI had significantly greater improvements in CVD risk parameters and reduced medication use and cost compared with those assigned to DSE. At 1 year, average number of medications prescribed to treat CVD risk factors was 3.1 +/- 1.8 for the ILI group and 3.6 +/- 1.8 for the DSE group (P < 0.0001), with estimated total monthly medication costs of $143 and $173, respectively (P < 0.0001). DSE participants meeting optimal care goals at 1 year were taking an average of 3.8 +/- 1.6 medications at an estimated cost of $194/month. ILI participants at optimal care required fewer medications (3.2 +/- 1.7) at lower cost ($154/month) (P < 0.001). At 1 year, ILI significantly improved CVD risk factors, while at the same time reduced medication use and cost. Continued intervention and follow-up will determine whether these changes are maintained and reduce cardiovascular risk.


Comparison of the induction of immunoglobulin M and G antibodies in mice with purified pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates.
The nature and kinetics of the serum antibody response to pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates were studied in mice. Bovine serum albumin and diphtheria and tetanus toxoids were used as carrier proteins. The purified polysaccharides induced only immunoglobulin M (IgM) antibodies in thymus-bearing as well as congenic athymic (nude) mice. The polysaccharides covalently conjugated to proteins produced IgM and IgG antibodies in normal mice, but only IgM antibodies in nude mice. A second dose of the polysaccharide-protein conjugates resulted in a booster effect in the IgG response to the polysaccharides. Moreover, memory B-cells, generated after a primary injection with the polysaccharide-protein conjugates, could be triggered to the production of IgG antibodies after a second injection with the pure polysaccharides alone. These data indicate that the antibody response to the pure polysaccharides is thymus independent and that this response can be changed into a thymus-dependent response by covalent conjugation of the polysaccharide to a thymus-dependent protein.


Estimation and partitioning of polygenic variation captured by common SNPs for Alzheimer's disease, multiple sclerosis and endometriosis.
Common diseases such as endometriosis (ED), Alzheimer's disease (AD) and multiple sclerosis (MS) account for a significant proportion of the health care burden in many countries. Genome-wide association studies (GWASs) for these diseases have identified a number of individual genetic variants contributing to the risk of those diseases. However, the effect size for most variants is small and collectively the known variants explain only a small proportion of the estimated heritability. We used a linear mixed model to fit all single nucleotide polymorphisms (SNPs) simultaneously, and estimated genetic variances on the liability scale using SNPs from GWASs in unrelated individuals for these three diseases. For each of the three diseases, case and control samples were not all genotyped in the same laboratory. We demonstrate that a careful analysis can obtain robust estimates, but also that insufficient quality control (QC) of SNPs can lead to spurious results and that too stringent QC is likely to remove real genetic signals. Our estimates show that common SNPs on commercially available genotyping chips capture significant variation contributing to liability for all three diseases. The estimated proportion of total variation tagged by all SNPs was 0.26 (SE 0.04) for ED, 0.24 (SE 0.03) for AD and 0.30 (SE 0.03) for MS. Further, we partitioned the genetic variance explained into five categories by a minor allele frequency (MAF), by chromosomes and gene annotation. We provide strong evidence that a substantial proportion of variation in liability is explained by common SNPs, and thereby give insights into the genetic architecture of the diseases.


Choice of reference sequence and assembler for alignment of Listeria monocytogenes short-read sequence data greatly influences rates of error in SNP analyses.
The wide availability of whole-genome sequencing (WGS) and an abundance of open-source software have made detection of single-nucleotide polymorphisms (SNPs) in bacterial genomes an increasingly accessible and effective tool for comparative analyses. Thus, ensuring that real nucleotide differences between genomes (i.e., true SNPs) are detected at high rates and that the influences of errors (such as false positive SNPs, ambiguously called sites, and gaps) are mitigated is of utmost importance. The choices researchers make regarding the generation and analysis of WGS data can greatly influence the accuracy of short-read sequence alignments and, therefore, the efficacy of such experiments. We studied the effects of some of these choices, including: i) depth of sequencing coverage, ii) choice of reference-guided short-read sequence assembler, iii) choice of reference genome, and iv) whether to perform read-quality filtering and trimming, on our ability to detect true SNPs and on the frequencies of errors. We performed benchmarking experiments, during which we assembled simulated and real Listeria monocytogenes strain 08-5578 short-read sequence datasets of varying quality with four commonly used assemblers (BWA, MOSAIK, Novoalign, and SMALT), using reference genomes of varying genetic distances, and with or without read pre-processing (i.e., quality filtering and trimming). We found that assemblies of at least 50-fold coverage provided the most accurate results. In addition, MOSAIK yielded the fewest errors when reads were aligned to a nearly identical reference genome, while using SMALT to align reads against a reference sequence that is ∼0.82% distant from 08-5578 at the nucleotide level resulted in the detection of the greatest numbers of true SNPs and the fewest errors. Finally, we show that whether read pre-processing improves SNP detection depends upon the choice of reference sequence and assembler. In total, this study demonstrates that researchers should test a variety of conditions to achieve optimal results.


Using the phenoscape knowledgebase to relate genetic perturbations to phenotypic evolution.
The abundance of phenotypic diversity among species can enrich our knowledge of development and genetics beyond the limits of variation that can be observed in model organisms. The Phenoscape Knowledgebase (KB) is designed to enable exploration and discovery of phenotypic variation among species. Because phenotypes in the KB are annotated using standard ontologies, evolutionary phenotypes can be compared with phenotypes from genetic perturbations in model organisms. To illustrate the power of this approach, we review the use of the KB to find taxa showing evolutionary variation similar to that of a query gene. Matches are made between the full set of phenotypes described for a gene and an evolutionary profile, the latter of which is defined as the set of phenotypes that are variable among the daughters of any node on the taxonomic tree. Phenoscape's semantic similarity interface allows the user to assess the statistical significance of each match and flags matches that may only result from differences in annotation coverage between genetic and evolutionary studies. Tools such as this will help meet the challenge of relating the growing volume of genetic knowledge in model organisms to the diversity of phenotypes in nature. The Phenoscape KB is available at http://kb.phenoscape.org.


Gas-phase transformation of phosphatidylcholine cations to structurally informative anions via ion/ion chemistry.
Gas-phase transformation of synthetic phosphatidylcholine (PC) monocations to structurally informative anions is demonstrated via ion/ion reactions with doubly deprotonated 1,4-phenylenedipropionic acid (PDPA). Two synthetic PC isomers, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC(16:0/18:1)) and 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (PC(18:1/16:0)), were subjected to this ion/ion chemistry. The product of the ion/ion reaction is a negatively charged complex, [PC + PDPA - H](-). Collisional activation of the long-lived complex causes transfer of a proton and methyl cation to PDPA, generating [PC - CH3](-). Subsequent collisional activation of the demethylated PC anions produces abundant fatty acid carboxylate anions and low-abundance acyl neutral losses as free acids and ketenes. Product ion spectra of [PC - CH3](-) suggest favorable cleavage at the sn-2 position over the sn-1 due to distinct differences in the relative abundances. In contrast, collisional activation of PC cations is absent of abundant fatty acid chain-related product ions and typically indicates only the lipid class via formation of the phosphocholine cation. A solution phase method to produce the gas-phase adducted PC anion is also demonstrated. Product ion spectra derived from the solution phase method are similar to the results generated via ion/ion chemistry. This work demonstrates a gas-phase means to increase structural characterization of phosphatidylcholines via ion/ion chemistry.


Barrett's oesophagus.
Barrett's oesophagus represents the replacement of stratified squamous epithelium by metaplastic columnar epithelium for 3 cm of the distal oesophagus. Gastro-oesophageal reflux, which affects 40% of the adult population, is the principal aetiological factor. This results in predominantly acid but also bile reflux (due to duodenogastrooesophageal reflux) through the lower oesophageal sphincter, transient relaxation of which accounts for the main mechanism of reflux. Conventional Barrett's oesophagus is reported in 11-13% of patients with symptomatic reflux and short segment Barrett's oesophagus (< 3.0 cm) in 18%. Approximately 50% of these patients have recognised complications on presentation, eg, carcinoma (15%). The disparity between clinical symptoms and endoscopic severity is due to reduced oesophageal mucosal sensitivity as a consequence of prolonged mucosal acid exposure. These rather alarming figures combined with the knowledge that Barrett's oesophagus is a pre-malignant condition (the diagnosis is associated with a 25-130-fold increase of malignancy) may account for the substantial increase in junctional gastrooesophageal malignancies. Symptomatic Barrett's oesophagus should be managed with full-dose proton pump inhibitors, eg, lansoprazole. Anti-reflux surgery should be reserved for the medically fit patient with recurrent symptomatic relapse in the histological absence of premalignant change. There is no evidence suggesting that surgery can be used as a prophylactic measure against malignancy. Encouraging short-term results have been obtained with photodynamic therapy in the management of high-grade dysplasia. However, columnar epithelium has been found underlying the regenerated squamous epithelium, suggesting that life-long surveillance is warranted.


Who, What, Where: an analysis of private sector family planning provision in 57 low- and middle-income countries.
Family planning service delivery has been neglected; rigorous analyses of the patterns of contraceptive provision are needed to inform strategies to address this neglect. We used 57 nationally representative Demographic and Health Surveys in low- and middle-income countries (2000-2013) in four geographic regions to estimate need for contraceptive services, and examined the sector of provision, by women's socio-economic position. We also assessed method mix and whether women were informed of side effects. Modern contraceptive use among women in need was lowest in sub-Saharan Africa (39%), with other regions ranging from 64% to 72%. The private sector share of the family planning market was 37-39% of users across the regions and 37% overall (median across countries: 41%). Private sector users accessed medical providers (range across regions: 30-60%, overall mean: 54% and median across countries 23%), specialised drug sellers (range across regions: 31-52%, overall mean: 36% and median across countries: 43%) and retailers (range across regions: 3-14%, overall mean: 6% and median across countries: 6%). Private retailers played a more important role in sub-Saharan Africa (14%) than in other regions (3-5%). NGOs and FBOs served a small percentage. Privileged women (richest wealth quintile, urban residents or secondary-/tertiary-level education) used private sector services more than the less privileged. Contraceptive method types with higher requirements (medical skills) for provision were less likely to be acquired from the private sector, while short-acting methods/injectables were more likely. The percentages of women informed of side effects varied by method and provider subtype, but within subtypes were higher among public than private medical providers for four of five methods assessed. Given the importance of private sector providers, we need to understand why women choose their services, what quality services the private sector provides, and how it can be improved. However, when prioritising one of the two sectors (public vs. private), it is critical to consider the potential impact on contraceptive prevalence and equity of met need.


Prospective evaluation of the International Study Group for Liver Surgery definition of bile leak after a liver resection and the role of routine operative drainage: an international multicentre study.
The International Study Group for Liver Surgery (ISGLS) proposed a definition for bile leak after liver surgery. A multicentre international prospective study was designed to evaluate this definition. Data collected prospectively from 949 consecutive patients on specific datasheets from 11 international centres were collated centrally. Bile leak occurred in 69 (7.3%) of patients, with 31 (3.3%), 32 (3.4%) and 6 (0.6%) classified as grade A, B and C, respectively. The grading system of severity correlated with the Dindo complication classification system (P < 0.001). Hospital length of stay was increased when bile leak occurred, from a median of 7 to 15 days (P < 0.001), as was intensive care stay (P < 0.001), and both correlated with increased severity grading of bile leak (P < 0.001). 96% of bile leaks occurred in patients with intra-operative drains. Drain placement did not prevent subsequent intervention in the bile leak group with a 5-15 times greater risk of intervention required in this group (P < 0.001). The ISGLS definition of bile leak after liver surgery appears robust and intra-operative drain usage did not prevent the need for subsequent drain placement.


Cyber security education is as essential as "the three R's".
Smartphones have diffused rapidly across South African society and constitute the most dominant information and communication technologies in everyday use. That being so, it is important to ensure that all South Africans know how to secure their smart devices. Doing so requires a high level of security awareness and knowledge. As yet, there is no formal curriculum addressing cyber security in South African schools. Indeed, it seems to be left to universities to teach cyber security principles, and they currently only do this when students take computing-related courses. The outcome of this approach is that only a very small percentage of South Africans, i.e. those who take computing courses at university, are made aware of cyber security risks and know how to take precautions. In this paper we found that, because this group is overwhelmingly male, this educational strategy disproportionately leaves young South African women vulnerable to cyber-attacks. We thus contend that cyber security ought to be taught as children learn the essential "3 Rs"-delivering requisite skills at University level does not adequately prepare young South Africans for a world where cyber security is an essential skill. Starting to provide awareness and knowledge at primary school, and embedding it across the curriculum would, in addition to ensuring that people have the skills when they need them, also remove the current gender imbalance in cyber security awareness.


Fraction of exhaled nitric oxide values in childhood are associated with 17q11.2-q12 and 17q12-q21 variants.
The fraction of exhaled nitric oxide (Feno) value is a biomarker of eosinophilic airway inflammation and is associated with childhood asthma. Identification of common genetic variants associated with childhood Feno values might help to define biological mechanisms related to specific asthma phenotypes. We sought to identify the genetic variants associated with childhood Feno values and their relation with asthma. Feno values were measured in children age 5 to 15 years. In 14 genome-wide association studies (N = 8,858), we examined the associations of approximately 2.5 million single nucleotide polymorphisms (SNPs) with Feno values. Subsequently, we assessed whether significant SNPs were expression quantitative trait loci in genome-wide expression data sets of lymphoblastoid cell lines (n = 1,830) and were related to asthma in a previously published genome-wide association data set (cases, n = 10,365; control subjects: n = 16,110). We identified 3 SNPs associated with Feno values: rs3751972 in LYR motif containing 9 (LYRM9; P = 1.97 × 10(-10)) and rs944722 in inducible nitric oxide synthase 2 (NOS2; P = 1.28 × 10(-9)), both of which are located at 17q11.2-q12, and rs8069176 near gasdermin B (GSDMB; P = 1.88 × 10(-8)) at 17q12-q21. We found a cis expression quantitative trait locus for the transcript soluble galactoside-binding lectin 9 (LGALS9) that is in linkage disequilibrium with rs944722. rs8069176 was associated with GSDMB and ORM1-like 3 (ORMDL3) expression. rs8069176 at 17q12-q21, but not rs3751972 and rs944722 at 17q11.2-q12, were associated with physician-diagnosed asthma. This study identified 3 variants associated with Feno values, explaining 0.95% of the variance. Identification of functional SNPs and haplotypes in these regions might provide novel insight into the regulation of Feno values. This study highlights that both shared and distinct genetic factors affect Feno values and childhood asthma.