Identification of Isthmin 1 as a Novel Clefting and Craniofacial Patterning Gene in Humans.
Orofacial clefts are one of the most common birth defects, affecting 1-2 per 1000 births, and have a complex etiology. High-resolution array-based comparative genomic hybridization has increased the ability to detect copy number variants (CNVs) that can be causative for complex diseases such as cleft lip and/or palate. Utilizing this technique on 97 nonsyndromic cleft lip and palate cases and 43 cases with cleft palate only, we identified a heterozygous deletion of Isthmin 1 in one affected case, as well as a deletion in a second case that removes putative 3' regulatory information. Isthmin 1 is a strong candidate for clefting, as it is expressed in orofacial structures derived from the first branchial arch and is also in the same "synexpression group" as fibroblast growth factor 8 and sprouty RTK signaling antagonist 1a and 2, all of which have been associated with clefting. CNVs affecting Isthmin 1 are exceedingly rare in control populations, and Isthmin 1 scores as a likely haploinsufficiency locus. Confirming its role in craniofacial development, knockdown or clustered randomly interspaced short palindromic repeats/Cas9-generated mutation of isthmin 1 in Xenopus laevis resulted in mild to severe craniofacial dysmorphologies, with several individuals presenting with median clefts. Moreover, knockdown of isthmin 1 produced decreased expression of LIM homeobox 8, itself a gene associated with clefting, in regions of the face that pattern the maxilla. Our study demonstrates a successful pipeline from CNV identification of a candidate gene to functional validation in a vertebrate model system, and reveals Isthmin 1 as both a new human clefting locus as well as a key craniofacial patterning gene.


Child drowning prevention in the Philippines: the beginning of a conversation.
This study describes a process to explore factors which contribute to child-drowning deaths and allows the development of appropriate strategies to prevent similar deaths in a selected site in the Northern Philippines. Data collection techniques used in obtaining baseline data include: review of drowning mortality records; key informant interviews; focus group discussions; and community walk-throughs. Risk factors identified which could or did contribute to drowning events were: proximity to bodies of water; inadequate child supervision; lack of information/awareness of prevention strategies; and lack of drowning prevention programme(s). Measures on how to prevent drowning deaths were explored and initial interventions were implemented through a committee convened by the community. These interventions include: community education sessions; capability building measures; redesigning of community wells; development of playpens; and use of barriers. Community engagement is a crucial element in the development and implementation of any health programme. This study demonstrates that by engaging and working with the community action occurs, however, there is a need to conduct further evaluation activities to determine if the actions by the community continued beyond the project and have resulted in a decrease in drowning. One of the strengths of the process described is that it is culturally appropriate and site-specific and allows the community to find the solutions itself. Exploration and delivery of further projects in larger areas is required to reduce drowning in the Philippines. An imperative is the evaluation which will provide valuable information on whether barriers are a sustainable and acceptable means of prevention to the community in the long term.


Prognostic Factors for Poor Cognitive Development in Children Born Very Preterm or With Very Low Birth Weight: A Systematic Review.
Cognitive delay is the most common form of impairment among children born very preterm (VPT) at 32 weeks or less or with very low birth weight (VLBW) of 1250 g or less. It is important to identify factors that are robust predictors of long-term outcome because the ability to predict future prognosis will assist in health care and educational service planning and provision. To identify prognostic factors for poor cognitive development in children born VPT or with VLBW. A systematic review was conducted using MEDLINE, EMBASE, and PyscINFO databases to identify studies published between January 1, 1990, and June 1, 2014, reporting multivariable prediction models for neurodevelopment in VPT or VLBW children. Thirty-one studies comprising 98 risk factor models for cognitive outcome were identified. Two independent reviewers extracted key information on study design, outcome definition, risk factor selection, model development, and reporting and conducted a risk-of-bias assessment. There was evidence that male sex, nonwhite race/ethnicity, lower level of parental education, and lower birth weight were predictive of global cognitive impairment in children younger than 5 years. In older children, only the influence of parental education was sustained. Male sex was also predictive of language impairment in early infancy, but not in middle childhood. Gestational age was a poor predictor of cognitive outcome, probably because of a reduced discriminatory power in cohorts restricted to a narrow gestational age range. The prognostic value of neonatal brain injury was unclear; however, studies adopted mixed strategies for managing children with physical or neurosensory disability. The influence of perinatal risk factors on cognitive development of VPT or VLBW children appears to diminish over time as environmental factors become more important. It is difficult to isolate cognitive outcomes from motor and neurosensory impairment, and the strategy for dealing with untestable children has implications for risk prediction.


Iodine-125 as a protein label in immunology.
Iodine-125 has the relatively long half-life of 60 days, and emits 27.3- to 35.5-kev photon radiation, permitting counting in an ordinary well crystal scintillation counter with 30 to 40 percent efficiency. The emission of low-energy electrons having the same general energy range as tritium beta radiation electrons allows high-resolution autoradiography. These properties, together with the other advantages of radioactive iodine, make this nuclide a particularly satisfactory label for immunologically active proteins.


The floristic heterogeneity of the Pantanal and the occurrence of species with different adaptive strategies to water stress.
The Pantanal is characterised by a diversity of environments with areas ranging from periodic or permanent heavy flooding to areas with low flood levels, and even environments that never flood. Plant species which inhabit the floodplain are distributed in specific niches, with influence of various phytogeographic domains, including the Seasonal Semi-deciduous Forest, Amazon Rainforest, Cerrado and Chaco, as well rocky remnants, with a wide ecological span in their components. In intensely flooded areas, aquatic macrophytes are widely distributed, with their dynamics closely linked to time, depth and extent of flooding. Although the term "Pantanal" suggests a huge swamp-type wetland, water level variation during a seasonal cycle does not directly reach the root system of many plants. The landscape diversity of the Pantanal wetland is molded by the flood pulse, which interferes with the dynamics of plant communities. Therefore, the retraction and expansion of populations or communities is reflected in important ecological characteristics, considering the variety of morphological, anatomical and ecophysiological features of the species, whose phenotype is the result of a particular genotype. The present study discusses peculiar issues in the adaptation of species distributed in the Pantanal biome and underscores the importance of multidisciplinary approaches to obtain conclusive data on adaptive studies.


Guidelines for pre-clinical animal and cellular models of MuSK-myasthenia gravis.
Muscle-specific tyrosine kinase (MuSK) autoantibodies are the hallmark of a form of myasthenia gravis (MG) that can challenge the neurologist and the experimentalist. The clinical disease can be difficult to treat effectively. MuSK autoantibodies affect the neuromuscular junction in several ways. When added to muscle cells in culture, MuSK antibodies disperse acetylcholine receptor clusters. Experimental animals actively immunized with MuSK develop MuSK autoantibodies and muscle weakness. Weakness is associated with reduced postsynaptic acetylcholine receptor numbers, reduced amplitudes of miniature endplate potentials and endplate potentials, and failure of neuromuscular transmission. Similar impairments have been found in mice injected with IgG from MG patients positive for MuSK autoantibody (MuSK-MG). The active and passive models have begun to reveal the mechanisms by which MuSK antibodies disrupt synaptic function at the neuromuscular junction, and should be valuable in developing therapies for MuSK-MG. However, translation into new and improved treatments for patients requires procedures that are not too cumbersome but suitable for examining different aspects of MuSK function and the effects of potential therapies. Study design, conduct and analysis should be carefully considered and transparently reported. Here we review what has been learnt from animal and culture models of MuSK-MG, and offer guidelines for experimental design and conduct of studies, including sample size determination, randomization, outcome parameters and precautions for objective data analysis. These principles may also be relevant to the increasing number of other antibody-mediated diseases that are now recognized.


Closed-loop control of gamma oscillations in the amygdala demonstrates their role in spatial memory consolidation.
Gamma is a ubiquitous brain rhythm hypothesized to support cognitive, perceptual, and mnemonic functions by coordinating neuronal interactions. While much correlational evidence supports this hypothesis, direct experimental tests have been lacking. Since gamma occurs as brief bursts of varying frequencies and durations, most existing approaches to manipulate gamma are either too slow, delivered irrespective of the rhythm's presence, not spectrally specific, or unsuitable for bidirectional modulation. Here, we overcome these limitations with an approach that accurately detects and modulates endogenous gamma oscillations, using closed-loop signal processing and optogenetic stimulation. We first show that the rat basolateral amygdala (BLA) exhibits prominent gamma oscillations during the consolidation of contextual memories. We then boost or diminish gamma during consolidation, in turn enhancing or impairing subsequent memory strength. Overall, our study establishes the role of gamma oscillations in memory consolidation and introduces a versatile method for studying fast network rhythms in vivo.


Metabolic and metagenomic outcomes from early-life pulsed antibiotic treatment.
Mammalian species have co-evolved with intestinal microbial communities that can shape development and adapt to environmental changes, including antibiotic perturbation or nutrient flux. In humans, especially children, microbiota disruption is common, yet the dynamic microbiome recovery from early-life antibiotics is still uncharacterized. Here we use a mouse model mimicking paediatric antibiotic use and find that therapeutic-dose pulsed antibiotic treatment (PAT) with a beta-lactam or macrolide alters both host and microbiota development. Early-life PAT accelerates total mass and bone growth, and causes progressive changes in gut microbiome diversity, population structure and metagenomic content, with microbiome effects dependent on the number of courses and class of antibiotic. Whereas control microbiota rapidly adapts to a change in diet, PAT slows the ecological progression, with delays lasting several months with previous macrolide exposure. This study identifies key markers of disturbance and recovery, which may help provide therapeutic targets for microbiota restoration following antibiotic treatment.


Impact of combined estradiol and norethindrone therapy on visuospatial working memory assessed by functional magnetic resonance imaging.
Hormones regulate neuronal function in brain regions critical to cognition; however, the cognitive effects of postmenopausal hormone therapy are controversial. The goal was to evaluate the effect of postmenopausal hormone therapy on neural circuitry involved in spatial working memory. A randomized, double-blind, placebo-controlled crossover study was performed. The study was performed in a tertiary care university medical center. Ten healthy postmenopausal women of average age 56.9 yr were recruited. Volunteers were randomized to the order they received hormone therapy (5 microg ethinyl estradiol and 1 mg norethindrone acetate). Subjects received hormone therapy or placebo for 4 wk, followed by a 1-month washout period with no medications, and then received the other treatment for 4 wk. At the end of each 4-wk treatment period, a functional magnetic resonance imaging study was performed using a nonverbal (spatial) working memory task, the Visual Delayed Matching to Sample task. The effects of hormone therapy on brain activation patterns were compared with placebo. Compared with the placebo condition, hormone therapy was associated with a more pronounced activation in the prefrontal cortex (BA 44 and 45), bilaterally (P < 0.001). Hormone therapy was associated with more effective activation of a brain region critical in primary visual working memory tasks. The data suggest a functional plasticity of memory systems in older women that can be altered by hormones.


Multidetector CT of pancreatic ductal adenocarcinoma: Effect of tube voltage and iodine load on tumour conspicuity and image quality.
To compare a low-tube-voltage with or without high-iodine-load multidetector CT (MDCT) protocol with a normal-tube-voltage, normal-iodine-load (standard) protocol in patients with pancreatic ductal adenocarcinoma (PDAC) with respect to tumour conspicuity and image quality. Thirty consecutive patients (mean age: 66 years, men/women: 14/16) preoperatively underwent triple-phase 64-channel MDCT examinations twice according to: (i) 120-kV standard protocol (PS; 0.75 g iodine (I)/kg body weight, n = 30) and (ii) 80-kV protocol A (PA; 0.75 g I/kg, n = 14) or protocol B (PB; 1 g I/kg, n = 16). Two independent readers evaluated tumour delineation and image quality blindly for all protocols. A third reader estimated the pancreas-to-tumour contrast-to-noise ratio (CNR). Statistical analysis was performed with the Chi-square test. Tumour delineation was significantly better in PB and PA compared with PS (P = 0.02). The evaluation of image quality was similar for the three protocols (all, P > 0.05). The highest CNR was observed with PB and was significantly better compared to PA (P = 0.02) and PS (P = 0.0002). In patients with PDAC, a low-tube-voltage, high-iodine-load protocol improves tumour delineation and CNR leading to higher tumour conspicuity compared to standard protocol MDCT. • Low-tube-voltage high-iodine-load MDCT improves pancreatic cancer conspicuity compared to a standard protocol. • The pancreas-to-tumour attenuation difference increases significantly by reducing the tube voltage. • The radiation exposure dose decreases by reducing the tube voltage.


Psychosocial influences on prisoner suicide: a case-control study of near-lethal self-harm in women prisoners.
We examined the psychosocial influences on female prisoner suicide by carrying out a study of near-lethal self-harm. We interviewed 60 women prisoners who had recently engaged in near-lethal self-harm (cases) and 60 others who had never carried out near-lethal acts in prison (controls) from all closed female prison establishments in England and Wales, using mixed quantitative and qualitative methods. We gathered information on socio-demographic and criminological variables, life events and childhood trauma, exposure to suicidal behaviour, contributory and precipitating factors for near-lethal self-harm, social support and psychological characteristics. While socio-demographic factors were only modestly associated with near-lethal self-harm, being on remand, in single cell accommodation, and reporting negative experiences of imprisonment were strong correlates. Recent life events and past trauma, including different forms of childhood abuse, were also significantly associated with near-lethal self-harm, as were a family history of suicide and high scores on measures of depression, aggression, impulsivity and hostility, and low levels of self-esteem and social support. Our findings underline the importance of both individual and prison-related factors for suicide in custody, and hence the need for a comprehensive approach to suicide prevention in women's prisons. Given the multiple needs of female prisoners at-risk of self-harm and suicide, complex psychosocial interventions are likely to be required, including interventions for abused and bereaved women, and initiatives to improve staff-prisoner relationships and reduce bullying. The findings of this research may provide insights into factors leading to suicidal behaviour in other forensic and institutional settings, such as detention centres and psychiatric hospitals, and may assist in developing suicide prevention policies for prisoners and other at-risk populations.


Efficient dynamic importance sampling of rare events in one dimension.
Exploiting stochastic path-integral theory, we obtain by simulation substantial gains in efficiency for the computation of reaction rates in one-dimensional, bistable, overdamped stochastic systems. Using a well-defined measure of efficiency, we compare implementations of "dynamic importance sampling" (DIMS) methods to unbiased simulation. The best DIMS algorithms are shown to increase efficiency by factors of approximately 20 for a 5k(B)T barrier height and 300 for 9k(B)T, compared to unbiased simulation. The gains result from close emulation of natural (unbiased), instantonlike crossing events with artificially decreased waiting times between events that are corrected for in rate calculations. The artificial crossing events are generated using the closed-form solution to the most probable crossing event described by the Onsager-Machlup action. While the best biasing methods require the second derivative of the potential (resulting from the "Jacobian" term in the action, which is discussed at length), algorithms employing solely the first derivative do nearly as well. We discuss the importance of one-dimensional models to larger systems, and suggest extensions to higher-dimensional systems.


Change in risk factors for coronary heart disease during 10 years of a community intervention programme (North Karelia project).
A comprehensive community based programme to control cardiovascular diseases was started in North Karelia, Finland, in 1972. Reductions in smoking, serum cholesterol concentrations, and blood pressure were among the central intermediate objectives. The effect of the programme during the 10 year period 1972-82 was evaluated by examining independent random population samples at the outset (1972) and five (1977) and 10 (1982) years later both in the programme and in a matched reference area. Over 10 000 subjects were studied in 1972 and 1977 (participation rate about 90%) and roughly 8000 subjects in 1982 (participation rate about 80%). Analyses were conducted of the estimated effect of the programme on the risk factor population means by comparing the baseline and five year and 10 year follow up results in the age range 30-59 years. The effect of the programme (net reduction in North Karelia) at 10 years among the middle aged male population was estimated to be a 28% reduction in smoking (p less than 0.001), a 3% reduction in mean serum cholesterol concentration (p less than 0.001), a 3% fall in mean systolic blood pressure (p less than 0.001), and a 1% fall in mean diastolic blood pressure (p less than 0.05). Among the female population the reductions were respectively, 14% (NS), 1% (NS), 5% (p less than 0.001), and 2% (p less than 0.05). During the first five years of the project (1972-7) the programme effectively reduced the population mean values of the major coronary risk factors. At 10 years the effects had persisted for serum cholesterol concentrations and blood pressure and were increased for smoking.


Kronecker product approximation for preconditioning in three-dimensional imaging applications.
We derive Kronecker product approximations, with the help of tensor decompositions, to construct approximations of severely ill-conditioned matrices that arise in three-dimensional (3-D) image processing applications. We use the Kronecker product approximations to derive preconditioners for iterative regularization techniques; the resulting preconditioned algorithms allow us to restore 3-D images in a computationally efficient manner. Through examples in microscopy and medical imaging, we show that the Kronecker approximation preconditioners provide a powerful tool that can be used to improve efficiency of iterative image restoration algorithms.


Progressive lung cancer determined by expression profiling and transcriptional regulation.
Clinically, our ability to predict disease outcome for patients with early stage lung cancer is currently poor. To address this issue, tumour specimens were collected at surgery from non-small cell lung cancer (NSCLC) patients as part of the European Early Lung Cancer (EUELC) consortium. The patients were followed-up for three years post-surgery and patients who suffered progressive disease (PD, tumour recurrence, metastasis or a second primary) or remained disease-free (DF) during follow-up were identified. RNA from both tumour and adjacent-normal lung tissue was extracted from patients and subjected to microarray expression profiling. These samples included 36 adenocarcinomas and 23 squamous cell carcinomas from both PD and DF patients. The microarray data was subject to a series of systematic bioinformatics analyses at gene, network and transcription factor levels. The focus of these analyses was 2-fold: firstly to determine whether there were specific biomarkers capable of differentiating between PD and DF patients, and secondly, to identify molecular networks which may contribute to the progressive tumour phenotype. The experimental design and analyses performed permitted the clear differentiation between PD and DF patients using a set of biomarkers implicated in neuroendocrine signalling and allowed the inference of a set of transcription factors whose activity may differ according to disease outcome. Potential links between the biomarkers, the transcription factors and the genes p21/CDKN1A and Myc, which have previously been implicated in NSCLC development, were revealed by a combination of pathway analysis and microarray meta-analysis. These findings suggest that neuroendocrine-related genes, potentially driven through p21/CDKN1A and Myc, are closely linked to whether or not a NSCLC patient will have poor clinical outcome.


Relationship between Vascular Resistance and Sympathetic Nerve Fiber Density in Arterial Vessels in Children With Sleep Disordered Breathing.
Sleep disordered breathing in children is associated with increased blood flow velocity and sympathetic overactivity. Sympathetic overactivity results in peripheral vasoconstriction and reduced systemic vascular compliance, which increases blood flow velocity during systole. Augmented blood flow velocity is recognized to promote vascular remodeling. Importantly, increased vascular sympathetic nerve fiber density and innervation in early life plays a key role in the development of early-onset hypertension in animal models. Examination of sympathetic nerve fiber density of the tonsillar arteries in children undergoing adenotonsillectomy for Sleep disordered breathing will address this question in humans. Thirteen children scheduled for adenotonsillectomy to treat sleep disordered breathing underwent pupillometry, polysomnography, flow-mediated dilation, resting brachial artery blood flow velocity (velocity time integral), and platelet aggregation. The dorsal lingual artery (tonsil) was stained and immunofluorescence techniques used to determine sympathetic nerve fiber density. Sympathetic nerve fiber density was correlated with increased resting velocity time integral (r=0.63; P<0.05) and a lower Neuronal Pupillary Index (r=-0.71, P<0.01), as well as a slower mean pupillary constriction velocity (mean, r=-0.64; P<0.05). A faster resting velocity time integral was associated with a slower peak pupillary constriction velocity (r=-0.77; P<0.01) and higher platelet aggregation to collagen antigen (r=0.64; P<0.05). Slower mean and peak pupillary constriction velocity were associated with higher platelet aggregation scores (P<0.05; P<0.01, respectively). These results indicate that sympathetic activity is associated with change in both the function and structure of systemic vasculature in children with sleep disordered breathing.


Pre- and post-training session evaluation for interobserver agreement and diagnostic accuracy of probe-based confocal laser endomicroscopy for biliary strictures.
Current diagnostic modalities for indeterminate biliary strictures offer low accuracy. Probe-based confocal laser endomicroscopy (pCLE) permits microscopic assessment of mucosal structures by obtaining real-time high-resolution images of the mucosal layers of the gastrointestinal tract. Previously, an interobserver study demonstrated poor to fair agreement even among experienced confocal endomicroscopy operators. Our objective was to assess interobserver agreement and diagnostic accuracy upon completion of a pCLE training session. Forty de-identified pCLE video clips of indeterminate biliary strictures were sent to five endoscopists at four tertiary care centers for scoring. Observers subsequently attended a teaching session by an expert pCLE user that included 20 training clips and rescored the same pCLE video clips, which were randomized and renumbered. Pre-training interobserver agreement for all observers was 'fair' (Κ: 0.31, P-value: <0.0001) and diagnostic accuracy was 72% (55-80%). Post-training interobserver agreement for all observers was 'substantial' (Κ: 0.74, P-value: <0.0001) and diagnostic accuracy was 89% (80-95%). Using a paired t-test, we observed an increase of 17% (95% CI 7.6-26.4) in post-training diagnostic accuracy (t = 5.01, df = 4, P-value 0.007). Interobserver agreement and diagnostic accuracy improved after observers underwent training by an expert pCLE user with a specific sequence set. Users should participate in such training programs to maximize diagnostic accuracy of pCLE evaluation.


Continual updating and monitoring of clinical prediction models: time for dynamic prediction systems?
Clinical prediction models (CPMs) have become fundamental for risk stratification across healthcare. The CPM pipeline (development, validation, deployment, and impact assessment) is commonly viewed as a one-time activity, with model updating rarely considered and done in a somewhat ad hoc manner. This fails to address the fact that the performance of a CPM worsens over time as natural changes in populations and care pathways occur. CPMs need constant surveillance to maintain adequate predictive performance. Rather than reactively updating a developed CPM once evidence of deteriorated performance accumulates, it is possible to proactively adapt CPMs whenever new data becomes available. Approaches for validation then need to be changed accordingly, making validation a continuous rather than a discrete effort. As such, "living" (dynamic) CPMs represent a paradigm shift, where the analytical methods dynamically generate updated versions of a model through time; one then needs to validate the system rather than each subsequent model revision.


Life course socioeconomic position and incidence of mid-late life depression in China and England: a comparative analysis of CHARLS and ELSA.
Despite the growing prevalence of depression in the Chinese elderly, there is conflicting evidence of life course socioeconomic position (SEP) and depression onset in China, and whether this association is akin to that observed in Western societies. We compared incident risk of mid-late life depression by childhood and adulthood SEP in China and England, a country where mental health inequality is firmly established. Depression-free participants from the China Health and Retirement Longitudinal Study (N=8508) and the English Longitudinal Study of Ageing (N=6184) were studied over 4 years. Depressive symptoms were classified as incident cases using the Center for Epidemiologic Studies Depression Scale criteria. Associations between SEP (education, wealth, residence ownership and childhood/adolescent deprivation) and depression symptom onset were assessed using Cox proportional hazards models. In China, we also investigated children's government employment status as a SEP marker. Higher education and wealth predicted lower incidence of depression in both countries. The association with non-ownership of residence appeared stronger in England (HR 1.61, 95% CI 1.41 to 1.86) than in China (HR 1.11, 95% CI 0.95 to 1.29), while that with childhood/adolescent deprivation was stronger in China (HR 1.43, 95% CI 1.29 - 1.60) than in England (HR 1.33, 95% CI 0.92 to 1.92). Chinese adults whose children were employed in high-status government jobs, had lower rates of depression onset. Consistent findings from China and England demonstrate that SEP is a pervasive determinant of mid-late life depression in very diverse social contexts. Together with conventional measures of SEP, the SEP of children also affects the mental health of older Chinese.


Does theory influence the effectiveness of health behavior interventions? Meta-analysis.
To systematically investigate the extent and type of theory use in physical activity and dietary interventions, as well as associations between extent and type of theory use with intervention effectiveness. An in-depth analysis of studies included in two systematic reviews of physical activity and healthy eating interventions (k = 190). Extent and type of theory use was assessed using the Theory Coding Scheme (TCS) and intervention effectiveness was calculated using Hedges's g. Metaregressions assessed the relationships between these measures. Fifty-six percent of interventions reported a theory base. Of these, 90% did not report links between all of their behavior change techniques (BCTs) with specific theoretical constructs and 91% did not report links between all the specified constructs with BCTs. The associations between a composite score or specific items on the TCS and intervention effectiveness were inconsistent. Interventions based on Social Cognitive Theory or the Transtheoretical Model were similarly effective and no more effective than interventions not reporting a theory base. The coding of theory in these studies suggested that theory was not often used extensively in the development of interventions. Moreover, the relationships between type of theory used and the extent of theory use with effectiveness were generally weak. The findings suggest that attempts to apply the two theories commonly used in this review more extensively are unlikely to increase intervention effectiveness.


Bidirectional Associations between Acceptability of Violence and Intimate Partner Violence from Adolescence to Young Adulthood.
Beliefs about the acceptability of intimate partner violence (IPV) are associated with the perpetration of IPV among adolescents. However, minimal research has examined whether this association persists across time or whether there is a bidirectional association between acceptability of IPV and the perpetration of IPV. The purpose of the present study was to examine bidirectional associations between acceptability of IPV and the perpetration of IPV from adolescence into young adulthood. A sample of diverse high school students (N = 1,042; 56% female) from the Southwestern United States were assessed each year for six consecutive years. At each assessment, participants completed measures of the acceptability of IPV and psychological and physical IPV perpetration. The mean age of the sample at the first assessment was 15.09 (SD = .79). Structural equation modeling demonstrated that acceptability of male-to-female and female-to-male IPV were not consistent predictors of one's own IPV perpetration over time. In addition, minimal evidence was found for a bidirectional association between acceptability of IPV and one's own IPV perpetration over time. Moreover, minimal gender differences were evident and there were no differences based on race/ethnicity. Despite the stability of beliefs about the acceptability of IPV over time from adolescence to young adulthood, findings suggest that acceptability of IPV is not a robust predictor of one's own IPV perpetration during this developmental time period. The implications of targeting beliefs about IPV in prevention and intervention programs are discussed.


Simplified approach to surface-wave scattering by blazed dielectric gratings.
The scattering properties of dielectric gratings having asymmetric triangular profiles are determined by a simple approach that views the grating structure as a double set of parallel planes. The energy of an incident surface wave is then scattered in preferential directions that are consistent with the grating periodicity and with a Bragg condition. As a result, the triangular shape can be chosen so that scattering occurs mostly into the region above the grating or into that below it, thus making it possible to maximize the efficiency of beam couplers or other devices. This desirable blazing property can be achieved by satisfying simple design criteria that are obtained from the scattering approach presented here. The range of grating parameters for strong blazing is derived, and gratings with trapezoidal profiles are also discussed.


Yeast Ypt11 is targeted to recycling endosomes in mammalian cells.
In yeast, Ypt11 or Ypt32 along with the highly homologous Ypt8 or Ypt31 has been reported to be an essential component of intra-Golgi trafficking and has been implicated in the budding of vesicles from the most distal Golgi compartment. In the present study, we show that, in human cells, after heterologous expression of GFP-Ypt11 (where GFP stands for green fluorescent protein), the protein is targeted to transferrin-positive recycling endosomes. This compartment has been shown to form extensive tubular networks on applying the drug Brefeldin A. We also show, by confocal fluorescent microscopy, that these networks also contain Rab11 in cells expressing CFP-Rab11a (where CFP stands for cyan fluorescent protein) fusion protein and that these structures are identical with those targeted by GFP-Ypt11.


Diagnosis of non-viral sexually transmitted infections in Lithuania and international recommendations.
The aim of this study is to evaluate the range, quality and availability of diagnostic services for non-viral sexually transmitted infections (STIs), i.e. C. trachomatis, N. gonorrhoeae, T. vaginalis and T. pallidum, in Lithuania from September 2002 to December 2003. Surveillance data describing the organisation and performance characteristics of non-viral STI diagnostic services in Lithuania were collected using a questionnaire and subsequent site-visits. International evidence-based recommendations for non-viral STI diagnosis were used to evaluate the quality of the STI diagnostics. There were 171 facilities providing non-viral STI diagnostic services for the 3.5 million inhabitants of Lithuania. However, only 6% (n=9) of the respondents (n=153) could provide a confirmatory diagnosis, in accordance with international recommendations, for the full minimum range of relevant non-viral STIs in Lithuania, i.e. C. trachomatis, N. gonorrhoeae, T. pallidum, and T. vaginalis. In addition, accessibility to STI diagnostic services differed significantly among the different counties in Lithuania. Several of the respondents analysed low numbers of samples each year, and overall the sampling size was extremely low, especially for C. trachomatis diagnostics. In Lithuania, optimisation of non-viral STI diagnostics as well as of epidemiological surveillance and management of STIs is crucial. It may be worth considering a decrease in the number of laboratories, with those remaining having the possibility of performing STI diagnostic services that are optimised, in concordance with international recommendations, standardised, and quality assured using systematic internal and external quality controls and systems. In addition, establishment of national inter-laboratory networks and reference centres for non-viral STIs is recommended.


Selective pressures for accurate altruism targeting: evidence from digital evolution for difficult-to-test aspects of inclusive fitness theory.
Inclusive fitness theory predicts that natural selection will favour altruist genes that are more accurate in targeting altruism only to copies of themselves. In this paper, we provide evidence from digital evolution in support of this prediction by competing multiple altruist-targeting mechanisms that vary in their accuracy in determining whether a potential target for altruism carries a copy of the altruist gene. We compete altruism-targeting mechanisms based on (i) kinship (kin targeting), (ii) genetic similarity at a level greater than that expected of kin (similarity targeting), and (iii) perfect knowledge of the presence of an altruist gene (green beard targeting). Natural selection always favoured the most accurate targeting mechanism available. Our investigations also revealed that evolution did not increase the altruism level when all green beard altruists used the same phenotypic marker. The green beard altruism levels stably increased only when mutations that changed the altruism level also changed the marker (e.g. beard colour), such that beard colour reliably indicated the altruism level. For kin- and similarity-targeting mechanisms, we found that evolution was able to stably adjust altruism levels. Our results confirm that natural selection favours altruist genes that are increasingly accurate in targeting altruism to only their copies. Our work also emphasizes that the concept of targeting accuracy must include both the presence of an altruist gene and the level of altruism it produces.


Prefrontal hemodynamic after-effects caused by rebreathing may predict affective states - A multimodal functional near-infrared spectroscopy study.
Brain activity has been shown to be influenced by respiratory behavior. Here, we evaluated whether respiration-induced hypo- or hypercapnia may support differentiation between physiological versus pathological respiratory behavior. In particular, we investigated whether systemic physiological measures could predict the brain's time-frequency hemodynamics after three respiratory challenges (i.e., breath-holding, rebreathing, and hyperventilation) compared to resting-state. Prefrontal hemodynamics were assessed in healthy subjects (N = 27) using functional near-infrared spectroscopy (fNIRS). Systemic physiological measures were assessed in form of heart rate, partial end-tidal carbon dioxide, respiration rate, and saturation of peripheral oxygen. Time-frequency dynamics were quantified using the wavelet transform coherence (i.e., defined here as cortical-systemic coherence). We found that the three respiratory challenges modulated cortical-systemic coherence differently: (1) After rebreathing, cortical-systemic coherence could be predicted from the amplitude of the heart rate (strong negative correlation). (2) After breath-holding, the same observation was made (moderate negative correlation). (3) After hyperventilation, no significant effect was observed. (4) These effects were found only in the frequency range of very low-frequency oscillations. The presented findings highlight a distinct role of rebreathing in predicting cortical-systemic coupling based on heart rate changes, which may represents a measure of affective states in the brain. The applied multimodal assessment of hemodynamic and systemic physiological measures during respiratory challenges may therefore have potential applications in the differentiation between physiological and pathological respiratory behavior.


Preclinical pharmacokinetic and toxicological evaluation of MIF-1 peptidomimetic, PAOPA: examining the pharmacology of a selective dopamine D2 receptor allosteric modulator for the treatment of schizophrenia.
Schizophrenia is a mental illness characterized by a breakdown in cognition and emotion. Over the years, drug treatment for this disorder has mainly been compromised of orthosteric ligands that antagonize the active site of the dopamine D2 receptor. However, these drugs are limited in their use and often lead to the development of adverse movement and metabolic side effects. Allosteric modulators are an emerging class of therapeutics with significant advantages over orthosteric ligands, including an improved therapeutic and safety profile. This study investigates our newly developed allosteric modulator, PAOPA, which is a specific modulator of the dopamine D2 receptor. Previous studies have shown PAOPA to attenuate schizophrenia-like behavioral abnormalities in preclinical models. To advance this newly developed allosteric drug from the preclinical to clinical stage, this study examines the pharmacokinetic behavior and toxicological profile of PAOPA. Results from this study prove the effectiveness of PAOPA in reaching the implicated regions of the brain for therapeutic action, particularly the striatum. Pharmacokinetic parameters of PAOPA were found to be comparable to current market antipsychotic drugs. Necropsy and histopathological analyses showed no abnormalities in all examined organs. Acute and chronic treatment of PAOPA indicated no movement abnormalities commonly found with the use of current typical antipsychotic drugs. Moreover, acute and chronic PAOPA treatment revealed no hematological or metabolic abnormalities classically found with the use of atypical antipsychotic drugs. Findings from this study demonstrate a better safety profile of PAOPA, and necessitates the progression of this newly developed therapeutic for the treatment of schizophrenia.


Climate and Developmental Plasticity: Interannual Variability in Grapevine Leaf Morphology.
The shapes of leaves are dynamic, changing over evolutionary time between species, within a single plant producing different shaped leaves at successive nodes, during the development of a single leaf as it allometrically expands, and in response to the environment. Notably, strong correlations between the dissection and size of leaves with temperature and precipitation exist in both the paleorecord and extant populations. Yet, a morphometric model integrating evolutionary, developmental, and environmental effects on leaf shape is lacking. Here, we continue a morphometric analysis of >5,500 leaves representing 270 grapevines of multiple Vitis species between two growing seasons. Leaves are paired one-to-one and vine-to-vine accounting for developmental context, between growing seasons. Linear discriminant analysis reveals shape features that specifically define growing season, regardless of species or developmental context. The shape feature, a more pronounced distal sinus, is associated with the colder, drier growing season, consistent with patterns observed in the paleorecord. We discuss the implications of such plasticity in a long-lived woody perennial, such as grapevine (Vitis spp.), with respect to the evolution and functionality of plant morphology and changes in climate.


Do breast quadrants explain racial disparities in breast cancer outcomes?
Tumors of the inner quadrants of the breast are associated with poorer survival than those of the upper-outer quadrant. It is unknown whether racial differences in breast cancer outcomes are modified by breast quadrant, in addition to comparisons among Asian subgroups. Using the Surveillance, Epidemiology, and End Results database, we analyzed data among women diagnosed with non-metastatic invasive breast cancer between 1990 and 2014. We performed Cox proportional hazards regression models to assess the associations of race with breast cancer-specific survival and overall survival, stratified by breast quadrants. The models were adjusted for age, year of the diagnosis, tumor size, grade, histological type, tumor laterality, lymph node, estrogen receptor, progesterone receptor, and treatments. Among 454,154 patients (73.0% White, 10.0% Black, 7.8% Asian/PI, and 9.2% Hispanic), 54.3% had tumors diagnosed in the upper-outer quadrant of the breast. Asian/PI women were more likely than White to have tumors diagnosed in the nipple/central portion of the breast and were less likely to have diagnosed in the upper-outer quadrant (P < 0.001), despite a similar distribution of breast quadrant between Black, Hispanic, and White women. Compared with White women, the multivariable-adjusted hazard ratios of breast cancer-specific mortality were 1.41 (95% CI 1.37-1.44) in Black women, 0.82 (95% CI 0.79-0.85) in Asian women, and 1.05 (95% CI 1.02-1.09) in Hispanic women. Among Asian subgroups, Japanese American women had a lower risk of breast cancer-specific mortality (HR = 0.68, 95% CI 0.62-0.74) compared with White women. Overall survival was similar to breast cancer-specific survival in each race group. The race-associated risks did not vary significantly by breast quadrants for breast cancer-specific mortality and all-cause mortality. Differences in breast cancer survival by race could not be attributed to tumor locations. Understanding the cultural, biological, and lifestyle factors that vary between White, African American, and ethnic subgroups of Asian American women may help explain these survival differences.


The venomous cocktail of the vampire snail Colubraria reticulata (Mollusca, Gastropoda).
Hematophagy arose independently multiple times during metazoan evolution, with several lineages of vampire animals particularly diversified in invertebrates. However, the biochemistry of hematophagy has been studied in a few species of direct medical interest and is still underdeveloped in most invertebrates, as in general is the study of venom toxins. In cone snails, leeches, arthropods and snakes, the strong target specificity of venom toxins uniquely aligns them to industrial and academic pursuits (pharmacological applications, pest control etc.) and provides a biochemical tool for studying biological activities including cell signalling and immunological response. Neogastropod snails (cones, oyster drills etc.) are carnivorous and include active predators, scavengers, grazers on sessile invertebrates and hematophagous parasites; most of them use venoms to efficiently feed. It has been hypothesized that trophic innovations were the main drivers of rapid radiation of Neogastropoda in the late Cretaceous. We present here the first molecular characterization of the alimentary secretion of a non-conoidean neogastropod, Colubraria reticulata. Colubrariids successfully feed on the blood of fishes, throughout the secretion into the host of a complex mixture of anaesthetics and anticoagulants. We used a NGS RNA-Seq approach, integrated with differential expression analyses and custom searches for putative secreted feeding-related proteins, to describe in detail the salivary and mid-oesophageal transcriptomes of this Mediterranean vampire snail, with functional and evolutionary insights on major families of bioactive molecules. A remarkably low level of overlap was observed between the gene expression in the two target tissues, which also contained a high percentage of putatively secreted proteins when compared to the whole body. At least 12 families of feeding-related proteins were identified, including: 1) anaesthetics, such as ShK Toxin-containing proteins and turripeptides (ion-channel blockers), Cysteine-rich secretory proteins (CRISPs), Adenosine Deaminase (ADA); 2) inhibitors of primary haemostasis, such as novel vWFA domain-containing proteins, the Ectonucleotide pyrophosphatase/phosphodiesterase family member 5 (ENPP5) and the wasp Antigen-5; 3) anticoagulants, such as TFPI-like multiple Kunitz-type protease inhibitors, Peptidases S1 (PS1), CAP/ShKT domain-containing proteins, Astacin metalloproteases and Astacin/ShKT domain-containing proteins; 4) additional proteins, such the Angiotensin-Converting Enzyme (ACE: vasopressive) and the cytolytic Porins. Colubraria feeding physiology seems to involve inhibitors of both primary and secondary haemostasis, anaesthetics, a vasoconstrictive enzyme to reduce feeding time and tissue-degrading proteins such as Porins and Astacins. The complexity of Colubraria venomous cocktail and the divergence from the arsenal of the few neogastropods studied to date (mostly conoideans) suggest that biochemical diversification of neogastropods might be largely underestimated and worth of extensive investigation.


A Fidelity-Embedded Regularization Method for Robust Electrical Impedance Tomography.
Electrical impedance tomography (EIT) provides functional images of an electrical conductivity distribution inside the human body. Since the 1980s, many potential clinical applications have arisen using inexpensive portable EIT devices. EIT acquires multiple trans-impedance measurements across the body from an array of surface electrodes around a chosen imaging slice. The conductivity image reconstruction from the measured data is a fundamentally ill-posed inverse problem notoriously vulnerable to measurement noise and artifacts. Most available methods invert the ill-conditioned sensitivity or the Jacobian matrix using a regularized least-squares data-fitting technique. Their performances rely on the regularization parameter, which controls the trade-off between fidelity and robustness. For clinical applications of EIT, it would be desirable to develop a method achieving consistent performance over various uncertain data, regardless of the choice of the regularization parameter. Based on the analysis of the structure of the Jacobian matrix, we propose a fidelity-embedded regularization (FER) method and a motion artifact reduction filter. Incorporating the Jacobian matrix in the regularization process, the new FER method with the motion artifact reduction filter offers stable reconstructions of high-fidelity images from noisy data by taking a very large regularization parameter value. The proposed method showed practical merits in experimental studies of chest EIT imaging.


Isolation of a strain of a Hantaan virus from a fatal case of hemorrhagic fever with renal syndrome in Slovenia.
Hemorrhagic fever with renal syndrome (HFRS) has been serologically confirmed in Slovenia during the last seven years. There is evidence that three hantaviruses (Hantaan, Puumala, and a newly described form termed Dobrava) circulate simultaneously in this area. Recently, a hantavirus was isolated from the urine and brain tissue of a fatal case of HFRS. Positive immunofluorescent reactions with reference human sera and monoclonal antibodies were first recognized after the second cell culture passage. Extensive cross-reactivity between our isolate and prototype Hantaan virus, strain 76-118, and Hantaan-like isolates from the former Yugoslavia, Fojnica and Plitvice, was revealed by enzyme-linked immunosorbent assay with specific rat antisera. The reaction pattern of the isolate was similar to the prototype Hantaan virus by indirect immunofluorescent assay with a panel of monoclonal antibodies. Furthermore, the specificity of the isolates was confirmed by analysis of polymerase chain reaction products of this virus with five restriction endonucleases. This appears to be the first isolation of a strain of prototype Hantaan virus from a fatal case of HFRS in Europe.


New evidence on racial and ethnic disparities in homeownership in the United States from 2001 to 2010.
Using 2001-2010 homeownership data for the United States we analyze changes in racial and ethnic disparities between whites and blacks, Asians, Mexicans, Cubans, Puerto Ricans and other Hispanics. We employ Integrated Public Use Microdata (IPUMS) combined with local credit scores and house price to income ratios. Controlling for demographic, income, wealth, employment, and housing characteristics, we find no significant differences between whites and Asians, Mexicans, or Cubans. Conversely, blacks and Puerto Ricans remain substantially disadvantaged. We conduct further analysis for the 2001-2003, 2004-2007, and 2008-2010 periods of the housing boom and collapse. Blacks and Puerto Ricans experienced decreased disparities during the peak years of the boom. Puerto Rican parity with whites continued to improve during the crash while gains among blacks eroded. The results suggest the homeownership differences between whites, Asians, Mexicans, and Cubans are apparently explained by socioeconomic status while racial disparities among blacks and Puerto Ricans evolved but continue to persist.


Inactivity of nitric oxide synthase gene in the atherosclerotic human carotid artery.
Nitric oxide (NO) inhibits thrombus formation, vascular contraction, and smooth muscle cell proliferation. We investigated whether NO release is enhanced after endothelial NO synthase (eNOS) gene transfer in atherosclerotic human carotid artery ex vivo. Western blotting and immunohistochemistry revealed that transduction enhanced eNOS expression; however, neither nitrite production nor NO release measured by porphyrinic microsensor was altered. In contrast, transduction enhanced NO production in non-atherosclerotic rat aorta and human internal mammary artery. In transduced carotid artery, calcium-dependent eNOS activity was minimal and did not differ from control conditions. Vascular tetrahydrobiopterin concentrations did not differ between the experimental groups. Treatment of transduced carotid artery with FAD, FMN, NADPH, L-arginine, and either sepiapterin or tetrahydrobiopterin did not alter NO release. Superoxide formation was similar in transduced carotid artery and control. Treatment of transduced carotid artery with superoxide dismutase (SOD), PEG-SOD, PEG-catalase did not affect NO release. eNOS transduction in atherosclerotic human carotid artery results in high expression without any measurable activity of the recombinant protein. The defect in the atherosclerotic vessels is neither caused by cofactor deficiency nor enhanced NO breakdown. Since angioplasty is performed in atherosclerotic arteries,eNOS gene therapy is unlikely to provide clinical benefit.


Exploring the feasibility and acceptability of the contents, design, and functionalities of an online intervention promoting mental health, wellbeing, and study skills in Higher Education students.
Substantial numbers of students in Higher Education (HE) are reporting mental health difficulties, such as mild to moderate symptoms of depression and anxiety. Coupled with academic skills challenges, these difficulties can lead to decreased academic performance, low levels of study satisfaction, and eventually drop out. Student support services are facing budget cuts and can only attend to limited numbers of students, usually the ones who present with more severe mental health problems. Moreover, face-to-face contact may not appeal to those students who feel embarrassed by their problems or are afraid of being stigmatised. To address this important problem, an online psychological wellbeing and study skills support system called MePlusMe, has been developed to provide personalised support to its users. In the present study we investigated the feasibility and acceptability of the contents, design, and functionalities of the system. An offline version of the system was introduced to 13 postgraduate and undergraduate students (mean age = 31.3 years, SD = 10.25 years; 4 males) in a UK HE Institution, who presented with mild or moderate mental health difficulties. The participants evaluated the design of the system, its functionalities, and contents at Baseline and at Weeks 2, 4, and 8. Participants found the system easy to use, professional, and efficient and its contents non-judgemental and informative. Participants stated that engaging with and practicing the techniques targeted at mental health difficulties led to improvements in positive thinking and self-confidence, while the study skills techniques were practical. Suggestions for further improvement included the development of an app and an option for direct engagement with professionals. The findings confirmed the acceptability of the contents, design and functionalities of the system, while providing useful information to inform its further development. Next steps include a feasibility study, which will test and quantify the effects on everyday functioning, mood, mental wellbeing, and academic self-efficacy after using the system, and subsequently a randomized controlled trial, which will evaluate its effectiveness.


The Effects of Subcellular Nanograting Geometry on the Formation and Growth of Bacterial Biofilms.
Biofilm formation by bacteria protects them against environmental stresses such as desiccation, shear forces and antimicrobial agents, making them much harder to remove and increasing their virulence and persistence in industrial water systems and biomedical equipment. One promising method of disrupting biofilm formation and growth is to employ passive surface structures to inhibit bacterial adhesion and aggregation. However, most studies thus far have mainly focused on the early stages of biofilm formation and it is unclear if the influence of surface topography in the early phase will propagate to later stages. Here, we attempt to address this with an investigation into the biofilm formation of Pseudomonas aeruginosa on 25 different nanograting geometries, with dimensions that were systematically varied from subcellular to cellular sizes. The biofilms were characterized from the exponential growth phase to the decline phase, in intervals of 24 H over 4 days, using confocal scanning laser microscopy. Comparing the maximum volume of biofilm formed on each surface over 96 H, it was found that approximately 1/3 of the nanograting geometries exhibited 72 ± 16 % lower biovolume density than a flat surface. Bacteria on these nanogratings were also observed to form 40 ± 11 % smaller microcolonies that were 17 ± 6 % less compact than that found on the control surface. The majority of these nanogratings had deep trenches (i.e. depth ≥ 70% of the cell diameter). Furthermore, P. aeruginosa cells were observed to multiply at approximately twice the rate on almost all the nanogratings compared to flat surfaces, but these cell populations also began to decline 24 H earlier than those on a flat surface. Using available literature on P. aeruginosa, a qualitative model was put forth, attributing the results to increased cell motility, decreased exopolysaccharide formation and disrupted psl adhesin/signal trails on nanogratings. These factors, together, led to the net effects of reduced attachment, increased scattering of cells and rapid decline of the biofilms on nanogratings. The insights derived from this study suggest that passive surface geometries can be designed and optimized to successfully control/inhibit biofilm formation and growth.


Lung cancer, cardiopulmonary mortality, and long-term exposure to fine particulate air pollution.
Associations have been found between day-to-day particulate air pollution and increased risk of various adverse health outcomes, including cardiopulmonary mortality. However, studies of health effects of long-term particulate air pollution have been less conclusive. To assess the relationship between long-term exposure to fine particulate air pollution and all-cause, lung cancer, and cardiopulmonary mortality. Vital status and cause of death data were collected by the American Cancer Society as part of the Cancer Prevention II study, an ongoing prospective mortality study, which enrolled approximately 1.2 million adults in 1982. Participants completed a questionnaire detailing individual risk factor data (age, sex, race, weight, height, smoking history, education, marital status, diet, alcohol consumption, and occupational exposures). The risk factor data for approximately 500 000 adults were linked with air pollution data for metropolitan areas throughout the United States and combined with vital status and cause of death data through December 31, 1998. All-cause, lung cancer, and cardiopulmonary mortality. Fine particulate and sulfur oxide--related pollution were associated with all-cause, lung cancer, and cardiopulmonary mortality. Each 10-microg/m(3) elevation in fine particulate air pollution was associated with approximately a 4%, 6%, and 8% increased risk of all-cause, cardiopulmonary, and lung cancer mortality, respectively. Measures of coarse particle fraction and total suspended particles were not consistently associated with mortality. Long-term exposure to combustion-related fine particulate air pollution is an important environmental risk factor for cardiopulmonary and lung cancer mortality.


Drivers of poor medical care.
The global ubiquity of overuse and underuse of health-care resources and the gravity of resulting harms necessitate an investigation of drivers to inform potential solutions. We describe the network of influences that contribute to poor care and suggest that it is driven by factors that fall into three domains: money and finance; knowledge, bias, and uncertainty; and power and human relationships. In each domain the drivers operate at the global, national, regional, and individual level, and are modulated by the specific contexts within which they act. We discuss in detail drivers of poor care in each domain.


Automated Interpretation of Blood Culture Gram Stains by Use of a Deep Convolutional Neural Network.
Microscopic interpretation of stained smears is one of the most operator-dependent and time-intensive activities in the clinical microbiology laboratory. Here, we investigated application of an automated image acquisition and convolutional neural network (CNN)-based approach for automated Gram stain classification. Using an automated microscopy platform, uncoverslipped slides were scanned with a 40× dry objective, generating images of sufficient resolution for interpretation. We collected 25,488 images from positive blood culture Gram stains prepared during routine clinical workup. These images were used to generate 100,213 crops containing Gram-positive cocci in clusters, Gram-positive cocci in chains/pairs, Gram-negative rods, or background (no cells). These categories were targeted for proof-of-concept development as they are associated with the majority of bloodstream infections. Our CNN model achieved a classification accuracy of 94.9% on a test set of image crops. Receiver operating characteristic (ROC) curve analysis indicated a robust ability to differentiate between categories with an area under the curve of >0.98 for each. After training and validation, we applied the classification algorithm to new images collected from 189 whole slides without human intervention. Sensitivity and specificity were 98.4% and 75.0% for Gram-positive cocci in chains and pairs, 93.2% and 97.2% for Gram-positive cocci in clusters, and 96.3% and 98.1% for Gram-negative rods. Taken together, our data support a proof of concept for a fully automated classification methodology for blood-culture Gram stains. Importantly, the algorithm was highly adept at identifying image crops with organisms and could be used to present prescreened, classified crops to technologists to accelerate smear review. This concept could potentially be extended to all Gram stain interpretive activities in the clinical laboratory.


Optical control of GPR40 signalling in pancreatic β-cells.
Fatty acids activate GPR40 and K+ channels to modulate β-cell function. Herein, we describe the design and synthesis of FAAzo-10, a light-controllable GPR40 agonist based on Gw-9508. FAAzo-10 is a potent GPR40 agonist in the trans-configuration and can be inactivated on isomerization to cis with UV-A light. Irradiation with blue light reverses this effect, allowing FAAzo-10 activity to be cycled ON and OFF with a high degree of spatiotemporal precision. In dissociated primary mouse β-cells, FAAzo-10 also inactivates voltage-activated and ATP-sensitive K+ channels, and allows us to control glucose-stimulated Ca2+ oscillations in whole islets with light. As such, FAAzo-10 is a useful tool to study the complex effects, with high specificity, which FA-derivatives such as Gw-9508 exert at multiple targets in mouse β-cells.


Combinatorial approaches to probe the sequence determinants of protein aggregation and amyloidogenicity.
Elucidation of the molecular determinants that drive proteins to aggregate is important both to advance our fundamental understanding of protein folding and misfolding, and as a step towards successful intervention in human disease. Combinatorial strategies enable unbiased and model-free approaches to probe sequence/structure relationships. Through the use of combinatorial methods, it is possible (i) to probe the sequence determinants of natural amyloid proteins by screening libraries of amino acid substitutions (mutations) to identify those that prevent amyloid formation; and (ii) to test new hypotheses about the mechanism of formation of amyloid fibrils by using these hypotheses to guide the design of combinatorial libraries of de novo amyloid-like proteins. Here, we review how these two approaches have been used to study the molecular determinants of protein aggregation and amyloidogenicity.


An optimized whole blood assay measuring expression and activity of NLRP3, NLRC4 and AIM2 inflammasomes.
The proinflammatory protease caspase-1 plays pivotal roles in central pathways of innate immunity, thereby contributing to pathogen clearance. Beside its physiological role, dysregulated activity of caspase-1 is known to contribute to an increasing number of diseases. In this study, we optimized and validated a low-volume human whole blood assay facilitating the measurement of caspase-1 activation and inflammasome-related gene expression upon stimulation of the NLRP3, NLRC4 or AIM2 inflammasome. Using the NLRP3 inflammasome specific inhibitor MCC950, we were able to measure the activity of canonical or alternative NLRP3 pathways, AIM2 and NLRC4 inflammasomes in whole blood. Based on our data we assume a superposition of NLRP3 and NLRC4 inflammasome activities in human whole blood following stimulation with S. typhimurium. The optimized whole blood assay may be suitable for diagnostic and research purposes for pediatric patients who can only donate small amounts of blood.


Adaptive evolution of the lager brewing yeast Saccharomyces pastorianus for improved growth under hyperosmotic conditions and its influence on fermentation performance.
An adaptive evolution method to obtain stable Saccharomyces pastorianus brewing yeast variants with improved fermentation capacity is described. The procedure involved selection for rapid growth resumption at high osmotic strength. It was applied to a lager strain and to a previously isolated ethanol-tolerant strain. Fermentation performance of strains was compared at 15 °P wort strength. A selected osmotolerant variant of the ethanol-tolerant strain showed significantly shorter fermentation time than the parent strain, producing 6.45% alcohol by volume beer in 4-5 days with mostly similar organoleptic properties to the original strain. Diacetyl and pentanedione contents were 50-75% and 3-methylbutyl acetate and 2-phenylethyl acetate 50% higher than with the original strain, leading to a small flavour change. The variant contained significantly less intracellular trehalose and glycogen than the parent. Transcriptional analysis of selected genes at 24 h revealed reduced transcription of hexose transport genes and increased transcription of the MALx1 and MALx2 genes, responsible for α-glucoside uptake and metabolism. It is suggested that an attenuated stress response contributes to the improved fermentation performance. Results show that sequential selection for both ethanol tolerance and rapid growth at high osmotic strength can provide strains with enhanced fermentation speed with acceptable product quality.


Inhibition of UVB-induced nonmelanoma skin cancer: a path from tea to caffeine to exercise to decreased tissue fat.
Oral administration of green tea, black tea, or caffeine (but not the decaffeinated teas) inhibited ultraviolet B radiation (UVB)-induced skin carcinogenesis in SKH-1 mice. Studies with caffeine indicated that its inhibitory effect on the ATR/Chk1 pathway is an important mechanism for caffeine's inhibition of UVB-induced carcinogenesis. The regular teas or caffeine increased locomotor activity and decreased tissue fat. In these studies, decreased dermal fat thickness was associated with a decrease in the number of tumors per mouse. Administration of caffeine, voluntary exercise, and removal of the parametrial fat pads all stimulated UVB-induced apoptosis, inhibited UVB-induced carcinogenesis, and stimulated apoptosis in UVB-induced tumors. These results suggest that caffeine administration, voluntary exercise, and removal of the parametrial fat pads inhibit UVB-induced carcinogenesis by stimulating UVB-induced apoptosis and by enhancing apoptosis in DNA-damaged precancer cells and in cancer cells. We hypothesize that tissue fat secretes antiapoptotic adipokines that have a tumor promoting effect.


A recurrent idiotype on monoclonal anti-human Ia antibodies.
We report a recurrent idiotype on a remarkably high fraction (4/19) of murine monoclonal antibodies specific for human Ia monomorphic determinants and elicited by separate immunizations. For three of them, the shared idiotype is associated with the antigen-combining site. These results indicate that the spectrum of mouse antibody responses to human Ia antigens may be based on recurrent idiotypes, suggesting a limited potential repertoire of murine monoclonal antibodies to human Ia antigens. Anti-idiotypic reagents might be helpful in dissecting this repertoire and to generate a mirror image of a human Ia antigenic map. Furthermore, antisera to the idiotype of antibodies specific for human Ia monomorphic determinants might help in elucidating the interactions between Ia molecules and receptors on immune cells.


The Early Awareness and Alert System in Sweden: History and Current Status.
Introduction: Over the past decades, early awareness and alert (EAA) activities and systems have gained importance and become a key early health technology assessment (HTA) tool. While a pioneer in HTA, Sweden had no national level EAA activities until 2010. We describe the evolution and current status of the Swedish EAA System. Methods: This was a historical analysis based on the knowledge and experience of the authors supplemented by a targeted review of published and gray literature as well as documents relating to EAA activities in Sweden. Key milestones and a description of the current state of the Swedish EAA System is presented. Results: Initiatives to establish a system for the identification and assessment of emerging health technologies in Sweden date back to the 1980s. In the 1990s, the Swedish Agency for HTA and Assessment of Social Services (SBU) supported the development of EuroScan as one of its founder members. In the mid-2000s, an independent regional initiative, driven by the Stockholm County Drug and Therapeutics Committee, resulted in the establishment of a regional horizon scanning function. By 2009, this work had expanded to a collaboration between the four biggest counties in Sweden. The following year it was further expanded to the national level and since then the Swedish EAA System has been carrying out identification, filtration and prioritization of new medicines, early assessment of the prioritized medicines, and dissemination of information. In 2015, the EAA System was incorporated into the Swedish national process for managed introduction and follow-up of new medicines. Outputs from the EAA System are now used to select new medicines for inclusion in this process. Conclusions: The Swedish EAA System started as a regional initiative and rapidly grew to become a national level activity. An important feature of the system today is its complete integration into the national process for managed introduction and follow-up of new medicines. The system will continue to evolve as a response both to the changing landscape of health innovations and to new policy initiatives at the regional, national and international level.


Modelling the arrival of invasive organisms via the international marine shipping network: a Khapra beetle study.
Species can sometimes spread significant distances beyond their natural dispersal ability by anthropogenic means. International shipping routes and the transport of shipping containers, in particular are a commonly recognised pathway for the introduction of invasive species. Species can gain access to a shipping container and remain inside, hidden and undetected for long periods. Currently, government biosecurity agencies charged with intercepting and removing these invasive species when they arrive to a county's border only assess the most immediate point of loading in evaluating a shipping container's risk profile. However, an invasive species could have infested a container previous to this point and travelled undetected before arriving at the border. To assess arrival risk for an invasive species requires analysing the international shipping network in order to identify the most likely source countries and the domestic ports of entry where the species is likely to arrive. We analysed an international shipping network and generated pathway simulations using a first-order Markov chain model to identify possible source ports and countries for the arrival of Khapra beetle (Trogoderma granarium) to Australia. We found Kaohsiung (Taiwan) and Busan (Republic of Korea) to be the most likely sources for Khapra beetle arrival, while the port of Melbourne was the most likely point of entry to Australia. Sensitivity analysis revealed significant stability in the rankings of foreign and Australian ports. This methodology provides a reliable modelling tool to identify and rank possible sources for an invasive species that could arrive at some time in the future. Such model outputs can be used by biosecurity agencies concerned with inspecting incoming shipping containers and wishing to optimise their inspection protocols.


A neglected disease of humans: a new focus of visceral leishmaniasis in Bakool, Somalia.
Visceral leishmaniasis (VL) was observed in children in Bakool region, Somalia, an area where VL has not been reported before. We describe the extent of the problem in this war- and famine-stricken area. A retrospective analysis was done of all cases admitted to a VL treatment centre between July 2000 and August 2001. Patients with longstanding fever, splenomegaly and a positive direct agglutination test (DAT; titre > 1:3200) were treated as suspected VL cases. A rapid epidemiological and entomological assessment was performed in the area. Species identification was attempted from blood samples by polymerase chain reaction-restriction fragment length polymorphism analysis of cysteine proteinase B genes. In 1 year, 230 serologically-positive cases were diagnosed as VL, and response to therapy was good in 91.6% of the 225 treated with sodium stibogluconate. Parasitological confirmation was attempted and obtained in 2 cases. Parasites were found to be most similar to Sudanese and Ethiopian reference strains of the Leishmania donovani complex. In a serological survey of 161 healthy displaced persons, 15% were positive by the leishmanin skin test and 3 (2%) were positive by the DAT. The sandfly captures showed Phlebotomus martini and P. vansomerenae. VL seems to be a longstanding and serious health problem in Bakool region. Food insecurity might have contributed to the emergence and detection of VL in this area.


Minimally Invasive Treatment of Biventricular Hydrocephalus Caused by a Giant Basilar Apex Aneurysm via a Staged Combination of Endoscopy and Endovascular Embolization: A Case Report.
Biventricular hydrocephalus caused by a Giant Basilar Apex Aneurysm (GBAA) is a rare finding that presents unique and challenging treatment decisions. We report a case of GBAA causing a life-threatening biventricular hydrocephalus in which both the aneurysm and hydrocephalus were given definitive treatment through a staged, minimally invasive approach. An obtunded 82-year-old male was found to have biventricular hydrocephalus caused by an unruptured GBAA obstructing the foramina of Monro. The patient was treated via staged, minimally invasive technique that first involved endoscopic fenestration of the septum pellucidum to create communication between the lateral ventricles. A programmable ventriculo-peritoneal shunt was then placed with a high-pressure setting. The patient was then loaded with dual anti-platelet therapy prior to undergoing endovascular coiling of the GBAA with adjacent stenting of the Posterior Cerebral Artery. He remained on dual anti-platelet therapy and the shunt setting was lowered at the bedside to treat the hydrocephalus. At 6-month follow up, the patient had returned to his cognitive baseline, speaking fluently and appropriately. Biventricular hydrocephalus caused by a GBAA can successfully be treated in a minimally invasive fashion utilizing a combination of endoscopy and endovascular therapy, even when a stent-assisted coiling is needed.


Chronic restraint stress down-regulates amygdaloid expression of polysialylated neural cell adhesion molecule.
The amygdala is a brain area which plays a decisive role in fear and anxiety. Since exposure to chronic stress can induce profound effects in emotion and cognition, plasticity in specific amygdaloid nuclei in response to prior stress has been hypothesized to account for stress-induced emotional alterations. In order to identify amygdala nuclei which may be affected under chronic stress conditions we evaluated the effects of 21-days chronic restraint stress on the expression of a molecule implicated crucially in alterations in structural plasticity: the polysialylated neural cell adhesion molecule. We found that polysialylated neural cell adhesion molecule-immunoreactivity within the amygdala, present in somata and neuronal processes, has a regional gradient with the central medial and medial amygdaloid nuclei showing the highest levels. Our results demonstrate that chronic restraint stress induced an overall reduction in polysialylated neural cell adhesion molecule-immunoreactivity in the amygdaloid complex, mainly due to a significant decrease in the central medial amygdaloid and medial amygdaloid nuclei. Our data suggest that polysialylated neural cell adhesion molecule in these nuclei may play a prominent role in functional and structural remodeling induced by stress, being a potential mechanism for cognitive and emotional modulation. Furthermore, these finding provide the first clear evidence that life experiences can regulate the expression of polysialylated neural cell adhesion molecule in the amygdaloid complex.


Dual Inhibition of Interleukin-23 and Interleukin-17 Offers Superior Efficacy in Mouse Models of Autoimmunity.
Therapies targeting either interleukin (IL)-23 or IL-17 have shown promise in treating T helper 17 (Th17)-driven autoimmune diseases. Although IL-23 is a critical driver of IL-17, recognition of nonredundant and independent functions of IL-23 and IL-17 has prompted the notion that dual inhibition of both IL-23 and IL-17 could offer even greater efficacy for treating autoimmune diseases relative to targeting either cytokine alone. To test this hypothesis, we generated selective inhibitors of IL-23 and IL-17 and tested the effect of either treatment alone compared with their combination in vitro and in vivo. In vitro, using a novel culture system of murine Th17 cells and NIH/3T3 fibroblasts, we showed that inhibition of both IL-23 and IL-17 completely suppressed IL-23-dependent IL-22 production from Th17 cells and cooperatively blocked IL-17-dependent IL-6 secretion from the NIH/3T3 cells to levels below either inhibitor alone. In vivo, in the imiquimod induced skin inflammation model, and in the myelin oligodendrocyte glycoprotein peptide-induced experimental autoimmune encephalomyelitis model, we demonstrated that dual inhibition of IL-17 and IL-23 was more efficacious in reducing disease than targeting either cytokine alone. Together, these data support the hypothesis that neutralization of both IL-23 and IL-17 may provide enhanced benefit against Th17 mediated autoimmunity and provide a basis for a therapeutic strategy aimed at dual targeting IL-23 and IL-17.


Does endotoxin-activated complement alter myocellular sodium homeostasis during sepsis?
Inappropriate complement activation is closely related to tissue injury and organ dysfunction during systemic infection. It is not clear, however, if endotoxin-induced complement activation is responsible for changes in myocellular sodium homeostasis during sepsis. Rats underwent cecal ligation and puncture (CLP) or sham operation. Twenty-four hours after operation, fast-twitch extensor digitorum longus (EDL) muscles were isolated, incubated at 30 degrees C for 1 hour in Krebs-Henseleit buffer (KHB) (pH 7.4), and used to measure intracellular Na+ and K+ contents. Blood samples were collected to measure serum hemolytic complement activity and endotoxin levels. In addition, EDL muscles isolated from normal animals were incubated at 30 degrees C for 1 hour with zymosan-activated (10 mg/mL at 37 degrees C for 1 hour) rat sera, with lipopolysaccharide (LPS)-activated (LPS from Escherichia coli 055:B5, 10 or 200 microg/mL at 37 degrees C for 30 minutes) rat sera, with heat-inactivated (56 degrees C for 30 minutes) rat sera, with LPS (1 or 20 microg/mL), or in KHB. EDL muscles isolated from normal animals were also incubated with septic sera collected 6 or 24 hours after CLP with or without administration of soluble complement receptor type 1 (20 mg/kg, intraperitoneally). Myocellular Na+ and K+ contents ([Na+]i and [K+]i) were assayed using "washout" technique. Soluble C5b-9 complex levels in zymosan-activated or LPS-activated human sera were determined by enzyme-linked immunosorbent assay to evaluate the degree of complement activation induced by zymosan or LPS. Myocellular [Na+]i and [Na+]i/[K+]i ratios increased significantly 24 hours after CLP as compared with sham operation and were associated with decreased serum hemolytic complement activity and increased serum endotoxin levels. Zymosan-activated rat sera at sublytic concentrations markedly increased [Na+]i and [Na+]i/[K+]i ratios in isolated EDL muscles relative to heat-inactivated rat sera. LPS-activated rat sera, however, did not alter these two indices. In addition, myocellular [Na+]i and [Na+]i/[K+]i ratios were equivalent among normal EDL muscles incubated with septic sera, soluble complement receptor type 1-treated septic sera, or KHB. These results collectively suggest that polymicrobial sepsis, as produced by CLP, alters sodium homeostasis in fast-twitch skeletal muscles in association with changes in systemic complement activation and circulating endotoxin levels. Although endotoxin can activate the complement cascade, endotoxin-induced complement activation does not appear to be responsible for changes in myocellular sodium homeostasis observed during sepsis in rats.


A proposed classification system for presigmoid approaches: a scoping review.
The "presigmoid corridor" covers a spectrum of approaches using the petrous temporal bone either as a target in treating intracanalicular lesions or as a route to access the internal auditory canal (IAC), jugular foramen, or brainstem. Complex presigmoid approaches have been continuously developed and refined over the years, leading to great heterogeneity in their definitions and descriptions. Owing to the common use of the presigmoid corridor in lateral skull base surgery, a simple anatomy-based and self-explanatory classification is needed to delineate the operative perspective of the different variants of the presigmoid route. Herein, the authors conducted a scoping review of the literature with the aim of proposing a classification system for presigmoid approaches. The PubMed, EMBASE, Scopus, and Web of Science databases were searched from inception to December 9, 2022, following the PRISMA Extension for Scoping Reviews guidelines to include clinical studies reporting the use of "stand-alone" presigmoid approaches. Findings were summarized based on the anatomical corridor, trajectory, and target lesions to classify the different variants of the presigmoid approach. Ninety-nine clinical studies were included for analysis, and the most common target lesions were vestibular schwannomas (60/99, 60.6%) and petroclival meningiomas (12/99, 12.1%). All approaches had a common entry pathway (i.e., mastoidectomy) but were differentiated into two main categories based on their relationship to the labyrinth: translabyrinthine or anterior corridor (80/99, 80.8%) and retrolabyrinthine or posterior corridor (20/99, 20.2%). The anterior corridor comprised 5 variations based on the extent of bone resection: 1) partial translabyrinthine (5/99, 5.1%), 2) transcrusal (2/99, 2.0%), 3) translabyrinthine proper (61/99, 61.6%), 4) transotic (5/99, 5.1%), and 5) transcochlear (17/99, 17.2%). The posterior corridor consisted of 4 variations based on the target area and trajectory in relation to the IAC: 6) retrolabyrinthine inframeatal (6/99, 6.1%), 7) retrolabyrinthine transmeatal (19/99, 19.2%), 8) retrolabyrinthine suprameatal (1/99, 1.0%), and 9) retrolabyrinthine trans-Trautman's triangle (2/99, 2.0%). Presigmoid approaches are becoming increasingly complex with the expansion of minimally invasive techniques. Descriptions of these approaches using the existing nomenclature can be imprecise or confusing. Therefore, the authors propose a comprehensive classification based on the operative anatomy that unequivocally describes presigmoid approaches simply, precisely, and efficiently.


Calumenin interacts with serum amyloid P component.
We recently reported the identification of human calumenin, a novel Ca(2+) binding, transformation-sensitive and secreted protein [Vorum et al. (1998) Biochim. Biophys. Acta 1386, 121-131; Vorum et al. (1999) Exp. Cell Res. 248, 473-481] belonging to the family of multiple EF-hand proteins of the secretory pathway that include reticulocalbin, ERC-55, Cab45 and crocalbin. In order to further investigate the extracellular functions of calumenin we immobilized the recombinant protein to a column. After application of a placental tissue extract we were able to elute one protein that interacts with calumenin in the presence of Ca(2+). Amino acid sequencing identified this protein as serum amyloid P component (SAP). Furthermore, we verified and characterized the calumenin-SAP interaction by the surface plasmon resonance technique. The findings indicate that calumenin may participate in the immunological defense system and could be involved in the pathological process of amyloidosis that leads to formation of amyloid deposits seen in different types of tissues.


Effect of Temperature-Sensitive Poloxamer Solution/Gel Material on Pericardial Adhesion Prevention: Supine Rabbit Model Study Mimicking Cardiac Surgery.
We investigated the mobility of a temperature-sensitive poloxamer/Alginate/CaCl2 mixture (PACM) in relation to gravity and cardiac motion and the efficacy of PACM on the prevention of pericardial adhesion in a supine rabbit model. A total of 50 rabbits were randomly divided into two groups according to materials applied after epicardial abrasion: PACM and dye mixture (group PD; n = 25) and saline as the control group (group CO; n = 25). In group PD, rabbits were maintained in a supine position with appropriate sedation, and location of mixture of PACM and dye was assessed by CT scan at the immediate postoperative period and 12 hours after surgery. The grade of adhesions was evaluated macroscopically and microscopically two weeks after surgery. In group PD, enhancement was localized in the anterior pericardial space, where PACM and dye mixture was applied, on immediate post-surgical CT scans. However, the volume of the enhancement was significantly decreased at the anterior pericardial space 12 hours later (P < .001). Two weeks after surgery, group PD had significantly lower macroscopic adhesion score (P = .002) and fibrosis score (P = .018) than did group CO. Inflammation score and expression of anti-macrophage antibody in group PD were lower than those in group CO, although the differences were not significant. In a supine rabbit model study, the anti-adhesion effect was maintained at the area of PACM application, although PACM shifted with gravity and heart motion. For more potent pericardial adhesion prevention, further research and development on the maintenance of anti-adhesion material position are required.


Detection of antigen-specific cytotoxic T lymphocytes: significance for investigative dermatology.
Novel recombinant tetrameric complexes of HLA class I molecules allow the direct visualization of antigen-specific CD8+ T cells using flow cytometry. By facilitating the quantification, isolation and phenotypic analysis of CD8+ T cells, the use of HLA tetramers has extended our understanding of the role of cellular immunity in various disease settings. Recently the technique has also been applied to the study of cutaneous disease and provides insights into mechanisms of dermatopathology.


The Pseudomonas aeruginosa PSL Polysaccharide Is a Social but Noncheatable Trait in Biofilms.
Extracellular polysaccharides are compounds secreted by microorganisms into the surrounding environment, and they are important for surface attachment and maintaining structural integrity within biofilms. The social nature of many extracellular polysaccharides remains unclear, and it has been suggested that they could function as either cooperative public goods or as traits that provide a competitive advantage. Here, we empirically tested the cooperative nature of the PSL polysaccharide, which is crucial for the formation of biofilms in Pseudomonas aeruginosa We show that (i) PSL is not metabolically costly to produce; (ii) PSL provides population-level benefits in biofilms, for both growth and antibiotic tolerance; (iii) the benefits of PSL production are social and are shared with other cells; (iv) the benefits of PSL production appear to be preferentially directed toward cells which produce PSL; (v) cells which do not produce PSL are unable to successfully exploit cells which produce PSL. Taken together, this suggests that PSL is a social but relatively nonexploitable trait and that growth within biofilms selects for PSL-producing strains, even when multiple strains are on a patch (low relatedness at the patch level).IMPORTANCE Many studies have shown that bacterial traits, such as siderophores and quorum sensing, are social in nature. This has led to an impression that secreted traits act as public goods, which are costly to produce but benefit both the producing cell and its surrounding neighbors. Theories and subsequent experiments have shown that such traits are exploitable by asocial cheats, but we show here that this does not always hold true. We demonstrate that the Pseudomonas aeruginosa exopolysaccharide PSL provides social benefits to populations but that it is nonexploitable, because most of the fitness benefits accrue to PSL-producing cells. Our work builds on an increasing body of work showing that secreted traits can have both private and public benefits to cells.


Defining Faecal Calprotectin Thresholds as a Surrogate for Endoscopic and Histological Disease Activity in Ulcerative Colitis-a Prospective Analysis.
Faecal calprotectin [FCal] levels are used as a surrogate marker for mucosal inflammation, but thresholds for defining endoscopic or histological disease activity in ulcerative colitis [UC] remain unclear. Using validated indices, prospective measurements of FCal, symptoms [Simple Colitis Clinical Activity Index, SCCAI], endoscopic [Ulcerative Colitis Endoscopic Index of Severity, UCEIS] and histological activity [Nancy index] were made over 6 months in patients enrolled into the TrueColours UC web-based monitoring programme. Repeated measurements correlation was performed between FCal and SCCAI, UCEIS, and Nancy indices using definitions for remission and active disease [UCEIS: remission ≤1, active ≥4; Nancy: remission ≤1, active ≥2; combined criteria: remission UCEIS ≤1 and Nancy ≤1, active UCEIS ≥4 and Nancy ≥2]. Receiver operating characteristic curves investigated FCal thresholds after maximising sensitivity for active disease. In 39 patients followed prospectively for 6 months, correlation coefficients between FCal and SCCAI, UCEIS, and Nancy indices were 0.271 (95% confidence interval [CI] 0.114-0.415), 0.741 [95% CI 0.289-0.922], and 0.876 [95% CI 0.605-0.965], respectively. Median FCal thresholds for remission using endoscopic, histological, or combined criteria were 71 μg/g [range 8-624], 91 μg/g [range 8-858], and 67 μg/g [range 8-479], respectively. The FCal threshold above which active disease was confirmed was 187 μg/g for UCEIS (area under the curve [AUC] 0.915), 72 μg/g for Nancy [AUC 0.824], and 187 μg/g for combined endoscopic and histological criteria [AUC 0.936]. Correlation between FCal and symptoms in UC is weak. In contrast, the correlation between FCal and endoscopic or histological activity is strong. An FCal ≥72 μg/g indicates histological inflammation [Nancy ≥2] and ≥187 μg/g indicates endoscopically active disease [UCEIS ≥4], whether combined with histopathology or not.


Eleven Cases of Hb J-Paris-I [HBA2: c.38C>A (or HBA1)]: A Stable α Chain Variant Elutes in the P3 Window on High-Performance Liquid Chromatography.
Hb J-Paris-I [HBA2: c.38C>A (or HBA1)] is a stable fast-moving hemoglobin (Hb) that elutes in the P3 window on high performance liquid chromatography (HPLC). The mutation can happen on either the α1- or α2-globin gene. Codon 12 changes from GCC to GAC to replace the alanine amino acid with aspartic acid. This change is external with no clinical significance. The elution in the P3 wave on HPLC can interfere with the glycated Hb assay by HPLC. In this study, data of 11 cases of Hb J-Paris-I were thoroughly presented. The majority of the cases were of Indian ethnicity. The mean value of Hb J-Paris-I on HPLC was 26.7 ± 2.0%. The retention time (RT) was 1.75 ± 0.03 min. The isoelectric focusing (IEF) mean value was -5.6 (range -6.1 to -4.9). Hb A2 was consistently reduced to 1.8 ± 0.3%. A fraction of 0.8% corresponding to the Hb A2-J-Paris-I (α2J-Paris-Iδ2) is likely to be concealed within the A0 peak of Hb A on HPLC. Interestingly, two cases were associated with two different polymorphisms [HBA2: c.-24C>G or Cap +14 (C>G) and HBA2: c.*136A>G polymorphism] without apparent effect on the variant expression.


Phenotypes and phorbol ester-induced differentiation of human histiocytic lymphoma cell lines (U-937 and SU-DHL-1) and Reed-Sternberg cells.
Hodgkin's mononuclear cells, Reed-Sternberg (H-RS) cells, and U-937 and SU-DHL-1 histocytic cell lines were induced to differentiate by phorbol ester in cultures. The phenotypes of cells were determined by a panel of antibodies specific for monocytes, histiocytes, and interdigitating reticulum cells. Before induction, SU-DHL-1 cells and H-RS cells expressed similar markers, such as HeFi-1, 2H9, 1A2, and 1E9. In addition, SU-DHL-1 cells were also stained by Tac and Leu M5. Other monocyte markers, including OK M1, Co Mo2, BRL Mo1, BRL Mo2, and Leu M3 were consistently negative in both types of cells. After induction, SU-DHL-1 cells conserved the same phenotype, but H-RS cells became negative for HeFi-1, 1A2, and 2H9. The U-937 cells expressed Leu M1 and Co Mo2 and became positive for Leu M5, OK M1, Co Mo2, BRL Mo2, 2H9, and 1E9 after phorbol ester induction. The U-937 cells did not express HeFi-1 or 1A2. The marker expression of H-RS cells, SU-DHL-1 cells, and U-937 cells were compared with those of histiocytes or interdigitating reticulum cells in lymphoid tissues and with neoplastic cells in true histiocytic lymphoma and malignant histiocytosis. It is concluded that SU-DHL-1, U-937, and H-RS cells are derived from or most closely related to fixed histiocytes, free histiocytes, and interdigitating reticulum cells, respectively. Our study further confirms the diagnosis of SU-DHL-1 as true histiocytic lymphoma but reveals that U-937 is a case of malignant histiocytosis rather than the previously diagnosed histiocytic lymphoma. The phenotypes and induction properties of SU-DHL-1 cells are quite different from those of U-937 cells, which suggests that true histiocytic lymphoma and malignant histiocytosis are two distinct disease entities.