Segregation analysis of apolipoproteins A-1 and B-100 measured before and after an exercise training program: the HERITAGE Family Study.
Complex segregation analyses of apolipoproteins (apo) A-1 and B-100 were performed in a sample of 520 individuals from 99 white families who participated in the HERITAGE Family Study. In these sedentary families, plasma apo A-1 and B-100 concentrations were measured before and after a 20-week endurance exercise training program. Baseline apo A-1 and B-100 were adjusted for the effects of age (age-adjusted baseline apo A-1 and B-100) and for the effects of age and BMI (age-BMI-adjusted baseline apo A-1 and B-100). The change in response to training was computed as a simple Delta (posttraining minus baseline) and was adjusted for age and the baseline (age-baseline-adjusted apo A-1 and B-100 responses to training). In the present study, a major gene could not be inferred for baseline apo A-1. Rather, we found a major effect along with a multifactorial effect accounting for 8% to 9% and 51% to 56% of the variance, respectively. In addition, no clear evidence supported a major-gene effect for its response to training, whereas the transmission of a major effect from parents to offspring was ambiguous, ie, genetic in nature or familial environmental in origin. The major effect accounted for 15% of the variance, with an additional 21% and 58% of the variance being accounted for by a multifactorial effect in parents and offspring, respectively. It is interesting to have obtained evidence of a putative recessive major locus for baseline apo B-100, which accounted for 50% to 56% of the variance, with an additional 25% to 29% of the variance due to a multifactorial effect. In contrast, no major effect for its response to training was identified, although a multifactorial effect was found that accounted for 27% of the variance. The novel findings arising from the present study are summarized as follows. Baseline apo A-1 and its response to training were influenced by a major effect and a multifactorial effect. Baseline apo B-100 was influenced by a putative major recessive gene with a multifactorial component, but its response to training was influenced solely by a multifactorial component in these sedentary families.


ACR guidance document on MR safe practices: 2013.
Because there are many potential risks in the MR environment and reports of adverse incidents involving patients, equipment and personnel, the need for a guidance document on MR safe practices emerged. Initially published in 2002, the ACR MR Safe Practices Guidelines established de facto industry standards for safe and responsible practices in clinical and research MR environments. As the MR industry changes the document is reviewed, modified and updated. The most recent version will reflect these changes.


Application of concentrated deep sea water inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice.
Mineral water from deep-sea bedrock, formed over thousands of years, is rich in minerals such as Ca, Mg, Na, K, Fe and others. Our present study was to investigate the preventive effects of natural deep-sea water on developing atopic dermatitis (AD). We elicited AD by application of DNCB (2,4-dinitro-chlorobezene) in Nc/Nga mouse dorsal skin. Deep Sea water (DSW) was filtered and concentrated by a nanofiltration process and reverse osmosis. We applied concentrated DSW (CDSW) to lesions five times per week for six weeks, followed by evaluation. 1% pimecrolimus ointment was used as positive control. The severity of skin lesions was assessed macroscopically and histologically. Levels of inflammatory mediators and cytokines in the serum were detected by Enzyme-linked immunosorbent assay (ELISA) and the levels of CD4+ and CD8+ spleen lymphocytes were determined by flow cytometry analysis. DNCB-treated mice showed atopic dermatitis-like skin lesions. Treatment of mice with CDSW reduced the severity of symptoms in the skin lesions, including edema, erythema, dryness, itching, and transepidermal water loss (TEWL). Histological analyses demonstrated that epidermal thickness and infiltration of inflammatory cells were decreased after CDSW treatment. Given these interesting observations, we further evaluated the effect of CDSW on immune responses in this AD model. Treatment AD mice with CDSW inhibited up-regulation of IgE, histamine, and pro-inflammatory cytokines in the serum. Also, the CD4+/CD8+ ratio in spleen lymphocyte was down-regulated after treatment with CDSW. Finally, cytokines, especially IL-4 and IL-10 which are important for Th2 cell development, were reduced. Our data suggests that topical application of CDSW could be useful in preventing the development of atopic dermatitis.


Death feigning as an adaptive anti-predator behaviour: Further evidence for its evolution from artificial selection and natural populations.
Death feigning is considered to be an adaptive antipredator behaviour. Previous studies on Tribolium castaneum have shown that prey which death feign have a fitness advantage over those that do not when using a jumping spider as the predator. Whether these effects are repeatable across species or whether they can be seen in nature is, however, unknown. Therefore, the present study involved two experiments: (a) divergent artificial selection for the duration of death feigning using a related species T. freemani as prey and a predatory bug as predator, demonstrating that previous results are repeatable across both prey and predator species, and (b) comparison of the death-feigning duration of T. castaneum populations collected from field sites with and without predatory bugs. In the first experiment, T. freemani adults from established selection regimes with longer durations of death feigning had higher survival rates and longer latency to being preyed on when they were placed with predatory bugs than the adults from regimes selected for shorter durations of death feigning. As a result, the adaptive significance of death-feigning behaviour was demonstrated in another prey-predator system. In the second experiment, wild T. castaneum beetles from populations with predators feigned death longer than wild beetles from predator-free populations. Combining the results from these two experiments with those from previous studies provided strong evidence that predators drive the evolution of longer death feigning.


Increasing the Efficiency of Spontaneous Sedimentation for the Diagnosis of Intestinal Protozoa.
The aim of this study was to evaluate the association between a longer sedimentation time, the reading of a larger number of slides, and the collection of multiple samples on the efficiency of the spontaneous sedimentation technique. Twenty-two patients with a previous parasitological exam positive for intestinal protozoa were recruited to collect new fecal samples (3 samples per patient) before the beginning of antiparasitic treatment. All collected fecal samples were used for spontaneous sedimentation and centrifuge-flotation techniques. Of these, all 22 patients were positive based on spontaneous sedimentation, and 59.1% (13/22) based on centrifuge flotation. The number of samples and the number of slides analyzed by spontaneous sedimentation influenced the number of positive cases. The modifications applied to the spontaneous sedimentation technique increased its performance in protozoa diagnosis.


2,4,6-Trinitrotoluene detection by laser-photofragmentation-laser-induced fluorescence.
Photofragmentation (PF) and subsequent nitric oxide (NO) laser-induced fluorescence (LIF) is being developed to measure the concentration of energetic materials (EM's) in soil and other media. Laser radiation near 226 nm photodissociates gas-phase EM to NO(2), which predissociates into NO that gives an intense luminescence. The EM concentration is inferred from the intensity of the NO fluorescence. We have studied the factors that affect the PF-LIF signal intensity, including the effect of buffer gas on the LIF spectrum of pure NO, the effect of 2,4,6-trinitrotoluene (TNT) pressure on the PF-LIF spectrum, the effect of buffer-gas pressure on the PF-LIF signal intensity of pure TNT, and the effect of temperature on the PF-LIF spectra of pure TNT and of TNT in simulated soil. Heating of the TNT sample above 343 K was found to increase the magnitude of the PF-LIF signal intensity significantly, but also was found to cause physical and chemical changes in the TNT sample. The effects of heating and evacuating on the TNT sample were investigated. TNT concentration calibration curves were obtained for TNT in simulated soil mixtures. The limit of detection of TNT in soil was estimated to be 40 parts in 10(9).


Change point estimation in monitoring survival time.
Precise identification of the time when a change in a hospital outcome has occurred enables clinical experts to search for a potential special cause more effectively. In this paper, we develop change point estimation methods for survival time of a clinical procedure in the presence of patient mix in a Bayesian framework. We apply Bayesian hierarchical models to formulate the change point where there exists a step change in the mean survival time of patients who underwent cardiac surgery. The data are right censored since the monitoring is conducted over a limited follow-up period. We capture the effect of risk factors prior to the surgery using a Weibull accelerated failure time regression model. Markov Chain Monte Carlo is used to obtain posterior distributions of the change point parameters including location and magnitude of changes and also corresponding probabilistic intervals and inferences. The performance of the Bayesian estimator is investigated through simulations and the result shows that precise estimates can be obtained when they are used in conjunction with the risk-adjusted survival time CUSUM control charts for different magnitude scenarios. The proposed estimator shows a better performance where a longer follow-up period, censoring time, is applied. In comparison with the alternative built-in CUSUM estimator, more accurate and precise estimates are obtained by the Bayesian estimator. These superiorities are enhanced when probability quantification, flexibility and generalizability of the Bayesian change point detection model are also considered.


Male spiders control offspring sex ratio through greater production of female-determining sperm.
Sex allocation theory predicts that when sons and daughters have different reproductive values, parents should adjust offspring sex ratio towards the sex with the higher fitness return. Haplo-diploid species directly control offspring sex ratio, but species with chromosomal sex determination (CSD) were presumed to be constrained by Mendelian segregation. There is now increasing evidence that CSD species can adjust sex ratio strategically, but the underlying mechanism is not well understood. One hypothesis states that adaptive control is more likely to evolve in the heterogametic sex through a bias in gamete production. We investigated this hypothesis in males as the heterogametic sex in two social spider species that consistently show adaptive female-biased sex ratio and in one subsocial species that is characterized by equal sex ratio. We quantified the production of male (0) and female (X) determining sperm cells using flow cytometry, and show that males of social species produce significantly more X-carrying sperm than 0-sperm, on average 70%. This is consistent with the production of more daughters. Males of the subsocial species produced a significantly lower bias of 54% X-carrying sperm. We also investigated whether inter-genomic conflict between hosts and their endosymbionts may explain female bias. Next generation sequencing showed that five common genera of bacterial endosymbionts known to affect sex ratio are largely absent, ruling out that endosymbiont bacteria bias sex ratio in social spiders. Our study provides evidence for paternal control over sex allocation through biased gamete production as a mechanism by which the heterogametic sex in CSD species adaptively adjust offspring sex ratio.


TFE3 rearrangements in adult renal cell carcinoma: clinical and pathologic features with outcome in a large series of consecutively treated patients.
Renal cell carcinoma (RCC) with chromosomal rearrangement of transcription factor for immunoglobulin heavy-chain enhancer 3 (TFE3) at Xp11.2 is a distinct subtype that was initially described in children and has been reported to display an indolent course. Recent reports have identified RCC with TFE3 rearrangements in adults and have suggested a more aggressive course in this population. However, only a few studies have examined these tumors in a large series of consecutively treated adults. We screened 632 RCCs from patients consecutively treated by surgery at a single institution by fluorescence in situ hybridization to detect TFE3 rearrangements. We identified 6 RCCs with TFE3 rearrangement. Patient ages ranged from 25 to 78 years and included 4 women and 2 men. Tumors showed significant histologic variability. Comparison of the clinical and pathologic features between RCCs with TFE3 rearrangements and RCCs without TFE3 rearrangements showed no significant differences. Follow-up period for patients with TFE3-rearranged RCC ranged from 0.8 to 16.5 years, with 4 of 6 dying from the disease. Cancer-specific survival for patients with TFE3-rearranged RCC was significantly worse than for patients with TFE3-rearrangement-negative papillary-type RCC (P<0.001) but not different from that for TFE3-rearrangement-negative clear cell-type RCC. In conclusion, we present an assessment of TFE3 rearrangement status in a large series of adults consecutively treated by surgery for RCC. Our findings confirm that RCCs with TFE3 rearrangement account for only approximately 1% of adult RCCs. The results also suggest that adult RCC with TFE3 rearrangement may be a clinically aggressive tumor.


Your Robot Therapist Will See You Now: Ethical Implications of Embodied Artificial Intelligence in Psychiatry, Psychology, and Psychotherapy.
Research in embodied artificial intelligence (AI) has increasing clinical relevance for therapeutic applications in mental health services. With innovations ranging from 'virtual psychotherapists' to social robots in dementia care and autism disorder, to robots for sexual disorders, artificially intelligent virtual and robotic agents are increasingly taking on high-level therapeutic interventions that used to be offered exclusively by highly trained, skilled health professionals. In order to enable responsible clinical implementation, ethical and social implications of the increasing use of embodied AI in mental health need to be identified and addressed. This paper assesses the ethical and social implications of translating embodied AI applications into mental health care across the fields of Psychiatry, Psychology and Psychotherapy. Building on this analysis, it develops a set of preliminary recommendations on how to address ethical and social challenges in current and future applications of embodied AI. Based on a thematic literature search and established principles of medical ethics, an analysis of the ethical and social aspects of currently embodied AI applications was conducted across the fields of Psychiatry, Psychology, and Psychotherapy. To enable a comprehensive evaluation, the analysis was structured around the following three steps: assessment of potential benefits; analysis of overarching ethical issues and concerns; discussion of specific ethical and social issues of the interventions. From an ethical perspective, important benefits of embodied AI applications in mental health include new modes of treatment, opportunities to engage hard-to-reach populations, better patient response, and freeing up time for physicians. Overarching ethical issues and concerns include: harm prevention and various questions of data ethics; a lack of guidance on development of AI applications, their clinical integration and training of health professionals; 'gaps' in ethical and regulatory frameworks; the potential for misuse including using the technologies to replace established services, thereby potentially exacerbating existing health inequalities. Specific challenges identified and discussed in the application of embodied AI include: matters of risk-assessment, referrals, and supervision; the need to respect and protect patient autonomy; the role of non-human therapy; transparency in the use of algorithms; and specific concerns regarding long-term effects of these applications on understandings of illness and the human condition. We argue that embodied AI is a promising approach across the field of mental health; however, further research is needed to address the broader ethical and societal concerns of these technologies to negotiate best research and medical practices in innovative mental health care. We conclude by indicating areas of future research and developing recommendations for high-priority areas in need of concrete ethical guidance.


Safety and efficacy of a feed additive consisting of Lacticaseibacillus rhamnosus CNCM I-3698 and Companilactobacillus sp. CNCM I-3699 for weaned piglets (STI Biotechnologie).
Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of Sorbiflore® ADVANCE, a feed additive consisting of Lacticaseibacillus rhamnosus CNCM I-3698 and Companilactobacillus sp. CNCM I-3699 intended to be used as a zootechnical additive (functional group: other zootechnical additives) in feed for weaned piglets to improve their performance. In a previous opinion, the additive was described as containing viable but not cultivable cells of the two strains in a 1:1 ratio, with a minimum of total lactic acid bacteria counts of 5 × 108 viable-forming units (VFU)/g additive. However, in that opinion, the Panel could not fully characterise the additive or conclude on its dermal/ocular irritancy or sensitisation potential. In the current assessment, the applicant provided supplementary information to address the missing information for the characterisation of the additive. The proposed methodology to discriminate and individually quantify the two strains composing the additive still presented limitations. Therefore, the Panel concluded that the data available do not allow to fully characterise the additive. The Panel was not in the position to conclude on the taxonomical identification of the strain CNCM I-3699, and consequently, on its eligibility for the application of the qualified presumption of safety (QPS) approach. Therefore, the previous conclusions on the safety of the additive based on the QPS approach could not be confirmed. The Panel was not in the position to conclude on the safety of the additive for the target species, consumer and the environment. Sorbiflore® ADVANCE is not irritant to skin. The Panel could not conclude on the eye irritancy or skin sensitisation potential of the additive.


The optimal sampling strategy for unfamiliar prey.
Precisely how predators solve the problem of sampling unfamiliar prey types is central to our understanding of the evolution of a variety of antipredator defenses, ranging from Müllerian mimicry to polymorphism. When predators encounter a novel prey item then they must decide whether to take a risk and attack it, thereby gaining a potential meal and valuable information, or avoid such prey altogether. Moreover, if predators initially attack the unfamiliar prey, then at some point(s) they should decide to cease sampling if evidence mounts that the type is on average unprofitable to attack. Here, I cast this problem as a "two-armed bandit," the standard metaphor for exploration-exploitation trade-offs. I assume that as predators encounter and attack unfamiliar prey they use Bayesian inference to update both their beliefs as to the likelihood that individuals of this type are chemically defended, and the probability of seeing the prey type in the future. I concurrently use dynamic programming to identify the critical informational states at which predator should cease sampling. The model explains why predators sample more unprofitable prey before complete rejection when the prey type is common and explains why predators exhibit neophobia when the unfamiliar prey type is perceived to be rare.


Epidemic and intervention modelling--a scientific rationale for policy decisions? Lessons from the 2009 influenza pandemic.
Outbreak analysis and mathematical modelling are crucial for planning public health responses to infectious disease outbreaks, epidemics and pandemics. This paper describes the data analysis and mathematical modelling undertaken during and following the 2009 influenza pandemic, especially to inform public health planning and decision-making. Soon after A(H1N1)pdm09 emerged in North America in 2009, the World Health Organization convened an informal mathematical modelling network of public health and academic experts and modelling groups. This network and other modelling groups worked with policy-makers to characterize the dynamics and impact of the pandemic and assess the effectiveness of interventions in different settings. The 2009 A(H1N1) influenza pandemic. Modellers provided a quantitative framework for analysing surveillance data and for understanding the dynamics of the epidemic and the impact of interventions. However, what most often informed policy decisions on a day-to-day basis was arguably not sophisticated simulation modelling, but rather, real-time statistical analyses based on mechanistic transmission models relying on available epidemiologic and virologic data. A key lesson was that modelling cannot substitute for data; it can only make use of available data and highlight what additional data might best inform policy. Data gaps in 2009, especially from low-resource countries, made it difficult to evaluate severity, the effects of seasonal variation on transmission and the effectiveness of non-pharmaceutical interventions. Better communication between modellers and public health practitioners is needed to manage expectations, facilitate data sharing and interpretation and reduce inconsistency in results.


The Science of Firescapes: Achieving Fire-Resilient Communities.
Wildland fire management has reached a crossroads. Current perspectives are not capable of answering interdisciplinary adaptation and mitigation challenges posed by increases in wildfire risk to human populations and the need to reintegrate fire as a vital landscape process. Fire science has been, and continues to be, performed in isolated "silos," including institutions (e.g., agencies versus universities), organizational structures (e.g., federal agency mandates versus local and state procedures for responding to fire), and research foci (e.g., physical science, natural science, and social science). These silos tend to promote research, management, and policy that focus only on targeted aspects of the "wicked" wildfire problem. In this article, we provide guiding principles to bridge diverse fire science efforts to advance an integrated agenda of wildfire research that can help overcome disciplinary silos and provide insight on how to build fire-resilient communities.


Genome-wide analysis of the basic leucine zipper (bZIP) transcription factor gene family in six legume genomes.
Plant bZIP proteins characteristically harbor a highly conserved bZIP domain with two structural features: a DNA-binding basic region and a leucine (Leu) zipper dimerization region. They have been shown to be diverse transcriptional regulators, playing crucial roles in plant development, physiological processes, and biotic/abiotic stress responses. Despite the availability of six completely sequenced legume genomes, a comprehensive investigation of bZIP family members in legumes has yet to be presented. In this study, we identified 428 bZIP genes encoding 585 distinct proteins in six legumes, Glycine max, Medicago truncatula, Phaseolus vulgaris, Cicer arietinum, Cajanus cajan, and Lotus japonicus. The legume bZIP genes were categorized into 11 groups according to their phylogenetic relationships with genes from Arabidopsis. Four kinds of intron patterns (a-d) within the basic and hinge regions were defined and additional conserved motifs were identified, both presenting high group specificity and supporting the group classification. We predicted the DNA-binding patterns and the dimerization properties, based on the characteristic features in the basic and hinge regions and the Leu zipper, respectively, which indicated that some highly conserved amino acid residues existed across each major group. The chromosome distribution and analysis for WGD-derived duplicated blocks revealed that the legume bZIP genes have expanded mainly by segmental duplication rather than tandem duplication. Expression data further revealed that the legume bZIP genes were expressed constitutively or in an organ-specific, development-dependent manner playing roles in multiple seed developmental stages and tissues. We also detected several key legume bZIP genes involved in drought- and salt-responses by comparing fold changes of expression values in drought-stressed or salt-stressed roots and leaves. In summary, this genome-wide identification, characterization and expression analysis of legume bZIP genes provides valuable information for understanding the molecular functions and evolution of the legume bZIP transcription factor family, and highlights potential legume bZIP genes involved in regulating tissue development and abiotic stress responses.


Bronchoalveolar lavage in children with cystic fibrosis: how many lobes should be sampled?
The European Respiratory Society guidance on bronchoalveolar lavage (BAL) in children was published in 2000. It recommended taking one BAL specimen from the most affected lobe or from the right middle lobe in diffuse disease. In 2007, the European Respiratory Society modified the recommendations for children with cystic fibrosis (CF), suggesting two BAL specimens (right middle lobe and the lingula or the most affected lobe). To determine if BAL samples from one or two lobes give the full picture of lower airway infection in children with CF. A retrospective review of all paediatric patients with CF who underwent flexible bronchoscopy between May 2007 and May 2009 was undertaken. As BAL specimens from all six lobes were collected, the BAL results were reviewed to identify if positive cultures would have been missed if only one lobe (right middle or most affected) or two lobes (right middle plus the lingula or most affected) had been sampled. The results of 39 bronchoscopic procedures in 31 children were reviewed. The BAL samples were taken from 6 lobes in all 39 procedures. Had only one lobe been used, 26 positive cultures (14 organisms) would have been missed in 11 patients. Had two lobes been used, 12 positive cultures (8 organisms) would have been missed in 7 patients. A single-lobe BAL is insufficient in assessing patients with CF for lower airway infection. Even when BAL specimens are taken from two lobes, a number of infections may be missed.


Impaired spatial working memory across saccades contributes to abnormal search in parietal neglect.
Visual neglect of left space following right parietal damage in humans involves a lateral bias in attention, apparent in many search tasks. We hypothesized that parietal neglect may also involve a failure to remember which locations have already been examined during visual search: an impairment in retaining searched locations across saccades. Using a new paradigm, we monitored gaze during search, while simultaneously probing whether observers judged they had found a new target, or judged instead that they were re-fixating a previously examined target. A patient with left neglect following focal right parietal infarction repeatedly re-fixated right locations. Critically, he often failed to remember that these locations had already been searched, treating old targets as new discoveries at an abnormal rate. In comparison, healthy age-matched control subjects rarely re-fixated targets, and mistook old targets as new targets even more rarely. The frequency of such mistakes in the parietal patient, for different conditions, correlated with the severity of his neglect. Control experiments indicated no perceptual localization deficit in non-search tasks. These results suggest a deficit in retaining searched locations across saccades in parietal neglect, in addition to the lateral spatial bias. Moreover, the former deficit exacerbates the latter, such that patients do not realize that the rightward locations favoured by their bias have already been examined during previous fixations and, for this reason, they saccade back to them repeatedly. The combination of the two deficits (a lateral bias plus a deficit in retaining locations already searched) may thus explain the pathological pattern of search that characterizes parietal neglect: why stimuli on the right are re-examined recursively, as if being searched for the first time, and hence why stimuli on the left continue to be ignored even with unlimited viewing time. These proposals accord with recent electrophysiological and functional imaging data, demonstrating posterior parietal involvement in the retention of target locations across saccades.


Mesoporous silica nanoparticles facilitating the dissolution of poorly soluble drugs in orodispersible films.
Orodispersible films (ODF) are immediately dissolving/disintegrating intraoral dosage forms, presented as substitutes of conventional tablets or capsules to ease problems associated with swallowing. Efforts have been made to be able to exploit ODFs as dosage forms for poorly soluble drugs. In the last two decades, mesoporous silica nanoparticles (MSNs) have been extensively used in drug delivery applications to overcome solubility problems of drugs. The tunable features of MSNs make them suitable candidates as drug carriers and solubility enhancers. In this study, the feasibility of MSNs as a carrier of poorly soluble drugs, using prednisolone as a model drug, in ODFs was investigated. Our results revealed that the increased amount of MSNs in ODFs leads to shortening of the disintegration time of the films. Drug content investigations showed that low dose ODFs with prednisolone incorporation efficiencies higher than 80% could be produced. Furthermore, the prednisolone release profile from ODFs can be tuned with the incorporation of MSNs as drug carrier (MSNpred). The MSNpred incorporated ODFs yield with immediate release of drug from the ODF, whereby 90% of the prednisolone content could be released in the first minutes. By modifying the MSNpred design with copolymer surface coating, prednisolone (cop-MSNpred) release can be modulated into a two-step sustained release profile. To sum up, the MSNs platform does not only provide careful low dose incorporation into ODF with high efficiency, but it also aids in tuning the drug release profiles from ODFs.


Malaria parasite development in mosquitoes.
Mosquitoes of the genus Anopheles transmit malaria parasites to humans. Anopheles mosquito species vary in their vector potential because of environmental conditions and factors affecting their abundance, blood-feeding behavior, survival, and ability to support malaria parasite development. In the complex life cycle of the parasite in female mosquitoes, a process termed sporogony, mosquitoes acquire gametocyte-stage parasites from blood-feeding on an infected host. The parasites carry out fertilization in the midgut, transform to ookinetes, then oocysts, which produce sporozoites. Sporozoites invade the salivary glands and are transmitted when the mosquito feeds on another host. Most individual mosquitoes that ingest gametocytes do not support development to the sporozoite stage. Bottle-necks occur at every stage of the cycle in the mosquito. Powerful new techniques and approaches exist for evaluating malaria parasite development and for identifying mechanisms regulating malaria parasite-vector interactions. This review focuses on those interactions that are important for the development of new approaches for evaluating and blocking transmission in nature.


Intestinal microbiota-derived metabolomic blood plasma markers for prior radiation injury.
Assessing whole-body radiation injury and absorbed dose is essential for remediation efforts following accidental or deliberate exposure in medical, industrial, military, or terrorist incidents. We hypothesize that variations in specific metabolite concentrations extracted from blood plasma would correlate with whole-body radiation injury and dose. Groups of C57BL/6 mice (n=12 per group) were exposed to 0, 2, 4, 8, and 10.4 Gy of whole-body gamma radiation. At 24 hours after treatment, all animals were euthanized, and both plasma and liver biopsy samples were obtained, the latter being used to identify a distinct hepatic radiation injury response within plasma. A semiquantitative, untargeted metabolite/lipid profile was developed using gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry, which identified 354 biochemical compounds. A second set of C57BL/6 mice (n=6 per group) were used to assess a subset of identified plasma markers beyond 24 hours. We identified a cohort of 37 biochemical compounds in plasma that yielded the optimal separation of the irradiated sample groups, with the most correlated metabolites associated with pyrimidine (positively correlated) and tryptophan (negatively correlated) metabolism. The latter were predominantly associated with indole compounds, and there was evidence that these were also correlated between liver and plasma. No evidence of saturation as a function of dose was observed, as has been noted for studies involving metabolite analysis of urine. Plasma profiling of specific metabolites related to pyrimidine and tryptophan pathways can be used to differentiate whole-body radiation injury and dose response. As the tryptophan-associated indole compounds have their origin in the intestinal microbiome and subsequently the liver, these metabolites particularly represent an attractive marker for radiation injury within blood plasma.


A differential scanning calorimetry study of retrieved orthopedic implants made of ultrahigh molecular weight polyethylene.
Differential scanning calorimetry (DSC) was used to examine thermal and thermooxidative properties of ultrahigh molecular weight polyethylene (UHMW-PE) of five acetabular components of failed orthopedic implants retrieved at revision of total hip arthoplasty. The results were compared with controls (unimplanted acetabular cups, a 20-year-old slab of UHMW-PE, and raw material). Profiles of exothermic peaks indicated increased levels of oxidation in all retrieved cups. In three retrieved cups, DSC revealed an additional peak of endotherm that was not seen in control samples. The additional endotherm peaks were not artifacts due to oxidation during scanning, heat buildup during cutting of the samples, or the sterilization method after retrieval. The additional peak was associated with the bulk of the polymer that was extracted with hexane. It varied in relative area, depending on its original location of the sample in a cup, implicating local variability in the extent of changes in material property. The distribution of the changes suggests that, during implantation, tissue exposure and friction affected the level of oxidation and degree of crystallinity in the UHMW-PE to a greater degree than did loading alone. Overall results showed that DSC may be a useful tool in evaluating changes in the properties of UHMW-PE orthopedic components in vivo.


Recent advances in understanding apicomplexan parasites.
Intracellular single-celled parasites belonging to the large phylum Apicomplexa are amongst the most prevalent and morbidity-causing pathogens worldwide. In this review, we highlight a few of the many recent advances in the field that helped to clarify some important aspects of their fascinating biology and interaction with their hosts. Plasmodium falciparum causes malaria, and thus the recent emergence of resistance against the currently used drug combinations based on artemisinin has been of major interest for the scientific community. It resulted in great advances in understanding the resistance mechanisms that can hopefully be translated into altered future drug regimens. Apicomplexa are also experts in host cell manipulation and immune evasion. Toxoplasma gondii and Theileria sp., besides Plasmodium sp., are species that secrete effector molecules into the host cell to reach this aim. The underlying molecular mechanisms for how these proteins are trafficked to the host cytosol ( T. gondii and Plasmodium) and how a secreted protein can immortalize the host cell ( Theileria sp.) have been illuminated recently. Moreover, how such secreted proteins affect the host innate immune responses against T. gondii and the liver stages of Plasmodium has also been unraveled at the genetic and molecular level, leading to unexpected insights. Methodological advances in metabolomics and molecular biology have been instrumental to solving some fundamental puzzles of mitochondrial carbon metabolism in Apicomplexa. Also, for the first time, the generation of stably transfected Cryptosporidium parasites was achieved, which opens up a wide variety of experimental possibilities for this understudied, important apicomplexan pathogen.


Cell adherence-promoted activity of Plesiomonas shigelloides groEL.
Previously, it has been demonstrated that the invasion of Caco-2 cells by Plesiomonas shigelloides induces apoptotic cell death. Therefore, the attachment to and colonization of eukaryotic intestinal host cells by P. shigelloides are important steps in causing pathogenicity. In this study, the participation of P. shigelloides GroEL in the attachment of P. shigelloides was examined. The groESL operon of P. shigelloides was isolated by PCR. The nucleotide sequence of the groESL operon of P. shigelloides revealed two ORFs of 294 nucleotides for groES and 1647 nucleotides for groEL. Cell fractionation and immunostaining experiments suggested that the GroEL of P. shigelloides was associated with the bacterial cell surface. The expression of the groEL gene was upregulated during the attachment and apoptosis-induction stages, and the expression of the protein was also induced during the attachment stage. Furthermore, GroEL efficiently promoted the attachment of P. shigelloides to Caco-2 cells, as measured by a FACSCalibur flow cytometer. These results demonstrated that GroEL has a positive influence on the attachment of P. shigelloides to Caco-2 cells.


The role of infections in the immunopathogensis of systemic sclerosis--evidence from serological studies.
Infections are believed to often play a role in the immunopathogenesis of autoimmune disorders; such is the case in systemic sclerosis (SSc). In order to evaluate the potential role infections may have on the pathogenesis of SSc, we assessed serological reactivity against various infectious agents in patients with SSc and compared them with healthy controls. Serological samples obtained from 80 patients with SSc were compared with 296 compatible healthy controls. Both groups were of European origin. All samples were tested for the presence of antibodies directed against hepatitis B virus, hepatitis C virus, toxoplasmosis, rubella, CMV, EBV, and Treponema pallidum. We applied Bio-Rad commercial and experimental kits to assess most antigens and ELISA assays to complete the panel. Patients with SSc had elevated IgM and IgG against Toxoplasma gondii and against CMV. Higher titers were also detected against the hepatitis B virus core protein (recombinant HBc antigen) using MONOLISA anti-HBc Plus commercial kit (Bio-Rad). A significantly higher rate of IgM antibodies against the capsid antigen of the EBV was detected in SSc patients compared with healthy controls, as well. These data demonstrate that antibodies against CMV, HBV, and toxoplasmosis were detected more often in patients with SSc. This association implies that infectious agents may have a role in disease pathogenesis and expression.


Helicobacter pylori VacA enhances prostaglandin E2 production through induction of cyclooxygenase 2 expression via a p38 mitogen-activated protein kinase/activating transcription factor 2 cascade in AZ-521 cells.
Treatment of AZ-521 cells with Helicobacter pylori VacA increased cyclooxygenase 2 (COX-2) mRNA in a time- and dose-dependent manner. A p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked elevation of COX-2 mRNA levels, whereas PD98059, which blocks the Erk1/2 cascade, partially suppressed the increase. Consistent with involvement of p38 MAPK, VacA-induced accumulation of COX-2 mRNA was reduced in AZ-521 cells overexpressing a dominant-negative p38 MAPK (DN-p38). Phosphatidylinositol-specific phospholipase C, which inhibits VacA-induced p38 MAPK activation, blocked VacA-induced COX-2 expression. In parallel with COX-2 expression, VacA increased prostaglandin E(2) (PGE(2)) production, which was inhibited by SB203580 and NS-398, a COX-2 inhibitor. VacA-induced PGE(2) production was markedly attenuated in AZ-521 cells stably expressing DN-p38. VacA increased transcription of a COX-2 promoter reporter gene and activated a COX-2 promoter containing mutated NF-kappaB or NF-interleukin-6 sites but not a mutated cis-acting replication element (CRE) site, suggesting direct involvement of the activating transcription factor 2 (ATF-2)/CREB-binding region in VacA-induced COX-2 promoter activation. The reduction of ATF-2 expression in AZ-521 cells transformed with ATF-2-small interfering RNA duplexes resulted in suppression of COX-2 expression. Thus, VacA enhances PGE(2) production by AZ-521 cells through induction of COX-2 expression via the p38 MAPK/ATF-2 cascade, leading to activation of the CRE site in the COX-2 promoter.


Dysmorphology at a distance: results of a web-based diagnostic service.
In 2007, the DYSCERNE pilot project funded by the European Commission Public Health Executive Agency (EU DG Sanco) aimed at setting up a network of expertise for patients with rare dysmorphic disorders. As part of DYSCERNE, a Dysmorphology Diagnostic System (DDS) was set up to enable clinicians throughout the EU to submit cases electronically for diagnosis using a secure, web-based interface, hosted at specified access points (Submitting nodes), in 26 different European countries. We report the outcome of this service for 200 cases submitted consecutively between January 2010 and 2012. Each case was reviewed by an average of five expert reviewers. An average of three possible syndromic diagnoses was suggested per case. In 22.5% of the cases, a consensus clinical diagnosis was reached. Genetic testing was suggested in 70.5% of the cases, whereas other laboratory investigations and diagnostic imaging were recommended in 35.5 and 26% of the cases, respectively. Further specialized opinions were suggested in 23.5% of the cases. Overall, a total of 181 very rare or extremely rare genetic syndromes were considered in the differential diagnosis of the 200 cases. In two cases, the reviewers suggested that the findings represented a new syndrome, and in one of these syndromes the underlying genetic cause was subsequently identified. Other benefits of the submission process included the possibility of directing the case submitters to specific centres for diagnostic testing or participation in research and educational benefit derived for both case submitters and reviewers.


Effect of core boundary curvature on the confinement losses of hollow antiresonant fibers.
We use numerical simulations to investigate how the curvature of the fiber core boundary influences the attenuation of hollow antiresonant fibers. We show the importance of a "negative" curvature core boundary in reducing confinement losses and also how, for certain curvatures, optical power is coupled resonantly to cladding modes. We simulate bending losses and find results in agreement with previously-reported experiments.


Source apportionment of airborne nanoparticles in a Middle Eastern city using positive matrix factorization.
Airborne nanoparticles have been studied worldwide, but little is known about their sources in the Middle East region, where hot, arid and dusty climatic conditions generally prevail. For the first time in Kuwait, we carried out size-resolved measurements of particle number distributions (PNDs) and concentrations (PNCs) in the 5-1000 nm size range. Measurements were made continuously for 31 days during the summer months of May and June 2013 using a fast-response differential mobility spectrometer (Cambustion DMS500) at a sampling rate of 10 Hz. Sources and their contributions were identified using the positive matrix factorization (PMF) approach that was applied to the PND data. Simultaneous measurements of gaseous pollutants (i.e., O3, NO, NOx, SO2 and CO), PM10, wind speed and direction were also carried out to aid the interpretation of the PMF results through the conditional probability function plots and Pearson product-moment correlations. Six major sources of PNCs were identified, contributing ∼46% (fresh traffic emissions), 27% (aged traffic emissions), 9% (industrial emissions), 9% (regional background), 6% (miscellaneous sources) and 3% (Arabian dust transport) of the total PNCs. The sources of nanoparticles and their PND profiles identified could serve as reference data to design more detailed field studies in the future and treat these sources in dispersion modelling and health impact assessment studies.


Maltreatment-associated psychiatric problems: an example of environmentally triggered ESSENCE?
This paper presents a new concept-maltreatment associated psychiatric problems (MAPP)-a syndrome of overlapping complex neurodevelopmental problems in children who have experienced abuse or neglect in early life. Children with MAPP are a hidden population in the community and, in clinical settings, their problems can seem overwhelming. Individual disorders associated with maltreatment are discussed as well as the overlap between these disorders and their shared environmental and genetic predisposing factors. Because of the complex and overlapping nature of MAPP, with symptoms emerging in early life, I argue that it should be considered an example of ESSENCE. Children presenting with likely MAPP should receive a comprehensive assessment, probing for symptoms of all of the ESSENCE disorders and leading to the use of evidence-based treatments where these are available.


Systolic blood pressure, socioeconomic status, and biobehavioral risk factors in a nationally representative US young adult sample.
In the National Longitudinal Study of Adolescent Health, a US longitudinal study of >15 000 young adults, we examined the extent to which socioeconomic status is linked to systolic blood pressure (SBP) and whether biobehavioral risk factors mediate the association. More than 62% of the participants had SBP >120 mm Hg and 12% had SBP >140 mm Hg. More than 66% were classified as at least overweight (body mass index >25 kg/m(2)), with >36% meeting criteria for at least class I obesity (body mass index >30 kg/m(2)). Multivariate models showed that higher household income and being married were independently associated with lower SBP. Higher body mass index, greater waist circumference, smoking, and higher alcohol intake were each independently associated with higher SBP. Meditational analyses suggested that higher education level was associated with lower SBP by way of lower body mass, smaller waist circumference, and lower resting heart rate. When these indirect effects were accounted for, education was not significantly associated with SBP. In contrast, household income remained associated with SBP even with control for all of the covariates. Results reinforce current public health concerns about rates of obesity and high blood pressure among young adults and suggest that disparities in education level and household income may play an important role in the observed decrements in health. Identifying modifiable mechanisms that link socioeconomic status to SBP using data from a large representative sample may improve risk stratification and guide the development of effective interventions.


Presynaptic and postsynaptic effects of histamine and histamine agonists in the superior cervical ganglion of the rat.
Extracellular and intracellular recording techniques were used to study the effects of histamine and the histamine agonists [impromidine (IMP) and 2-thiazolylethylamine (2-TH)] on synaptic transmission in the superior cervical ganglion of the rat in vitro. At the concentrations employed (up to 10(-5) M) these compounds did not produce detectable effects on the electrical properties of the postsynaptic neurons. Histamine produced a dose-dependent reduction in the amplitude of the extracellularly-recorded presynaptic and postsynaptic compound action potential. The H2 receptor agonist impromidine reduced only the postganglionic compound action potential. Cimetidine, a specific H2 receptor antagonist, produced parallel shifts in the log dose-response curves for impromidine. Impromidine also reduced the average size of the evoked excitatory postsynaptic potential. The reduction of the mean amplitude of the excitatory postsynaptic potential was due to a decrease in the amount of acetylcholine (ACh) liberated by each preganglionic volley (mean quantal content, m) and a diminution in quantal size. The H1 receptor agonist, 2-TH produced a dose-dependent increase in the presynaptic and postsynaptic compound action potential and in m. The increase in m was not associated with changes in quantal size. The H1 antagonists, pyrilamine and promethazine, did not prevent facilitation of ganglionic transmission induced by 2-TH. It is concluded that histamine H1 and H2 receptors exist on preganglionic axons, or terminals in sympathetic ganglia of the rat. Activation of H1 receptors facilitates release of ACh whereas H2 receptor activation results in depressed release.


Empirical assessment of analysis workflows for differential expression analysis of human samples using RNA-Seq.
RNA-Seq has supplanted microarrays as the preferred method of transcriptome-wide identification of differentially expressed genes. However, RNA-Seq analysis is still rapidly evolving, with a large number of tools available for each of the three major processing steps: read alignment, expression modeling, and identification of differentially expressed genes. Although some studies have benchmarked these tools against gold standard gene expression sets, few have evaluated their performance in concert with one another. Additionally, there is a general lack of testing of such tools on real-world, physiologically relevant datasets, which often possess qualities not reflected in tightly controlled reference RNA samples or synthetic datasets. Here, we evaluate 219 combinatorial implementations of the most commonly used analysis tools for their impact on differential gene expression analysis by RNA-Seq. A test dataset was generated using highly purified human classical and nonclassical monocyte subsets from a clinical cohort, allowing us to evaluate the performance of 495 unique workflows, when accounting for differences in expression units and gene- versus transcript-level estimation. We find that the choice of methodologies leads to wide variation in the number of genes called significant, as well as in performance as gauged by precision and recall, calculated by comparing our RNA-Seq results to those from four previously published microarray and BeadChip analyses of the same cell populations. The method of differential gene expression identification exhibited the strongest impact on performance, with smaller impacts from the choice of read aligner and expression modeler. Many workflows were found to exhibit similar overall performance, but with differences in their calibration, with some biased toward higher precision and others toward higher recall. There is significant heterogeneity in the performance of RNA-Seq workflows to identify differentially expressed genes. Among the higher performing workflows, different workflows exhibit a precision/recall tradeoff, and the ultimate choice of workflow should take into consideration how the results will be used in subsequent applications. Our analyses highlight the performance characteristics of these workflows, and the data generated in this study could also serve as a useful resource for future development of software for RNA-Seq analysis.


Perceptions Regarding Tobacco Cessation Counselling among Dental Students and Graduates: A Cross-Sectional Study.
Background: Tobacco usage is a global concern and it is essential to curb its usage and increase awareness among patients. Dental professionals' contribution in tobacco cessation will definitely make a stark difference. Thus it is important to assess dental students' and graduates' knowledge, attitude and practice towards Tobacco cessation counselling (TCC). Materials and Methods: The descriptive, cross-sectional survey was carried out among 286 students of a dental college in South India. A well-structured, pretested, self-administered questionnaire consisting of 17 close-ended questions was employed to assess knowledge, attitude and practice towards TCC. Results: Almost all study participants agreed that tobacco cessation counselling is under the scope of dental practice and admitted that it is the responsibility of dental professionals to educate patients for the same. Majority of graduates responded that they have done tobacco cessation counselling whereas less than two-third of undergraduates have done the same and difference between two groups was statistically significant (p<0.001). A significantly higher number of graduates (79%) were aware of 5A's and 5R's protocol for TCC as compared to undergraduates (50%). But less than one-fifth of study population have actually applied this protocol for TCC in clinical practice. Conclusion: There is a need to modify the dental curriculum and motivate young graduates and students about TCC. This will develop a more professional competence and helps to encourage the dental professionals in developing a preventive mind set about tobacco use. Thus, it will help in enhancing the long-term rates of quitting tobacco among patients, which will prove beneficial in controlling tobacco related diseases in near future.


Women's attitudes to condoms and female-controlled means of protection against HIV and STDs in south-western Uganda.
The consistent and correct use of the male condom makes it highly effective in both disease prevention and as a contraceptive method. However, it is also well recognized that its use is under men's control. Because of this vital limitation, there have been frequent calls for female-controlled methods of HIV prevention, particularly from women from sub-Saharan Africa. Here we report on data collected in focus-group discussions (FGDs) with women aged 17-54 in South-Western Uganda. A total of 138 women, from rural villages, urban family planning clinics and a truck-stop town, were recruited to participate in 18 FGDs on the male condom, the female condom and existing formulations of vaginal microbicidal products. Three themes emerged: (i) problems with men's control over the male condom, (ii) the importance of control over and secrecy about protective measures and (iii) sexual pleasure associated with different methods. We found that the female condom, while being perceived as an improvement over the male condom, was recognized as having limited value because of the need to agree its use prior to sex taking place. Other products were considered to be significantly better than the female condom; the sponge, in particular, was perceived as having advantages over every other product. Women like the fact that it could be inserted some time before, and left in place some time after, sexual intercourse, that it was effective for multiple instances of intercourse, and that men would be unaware that it was being employed. Female-controlled methods to prevent sexually transmitted infections, including HIV, and to increase reproductive choice, hold the promise of ceding some control over sexual and reproductive health to women.


The α2δ-1 subunit remodels CaV1.2 voltage sensors and allows Ca2+ influx at physiological membrane potentials.
Excitation-evoked calcium influx across cellular membranes is strictly controlled by voltage-gated calcium channels (CaV), which possess four distinct voltage-sensing domains (VSDs) that direct the opening of a central pore. The energetic interactions between the VSDs and the pore are critical for tuning the channel's voltage dependence. The accessory α2δ-1 subunit is known to facilitate CaV1.2 voltage-dependent activation, but the underlying mechanism is unknown. In this study, using voltage clamp fluorometry, we track the activation of the four individual VSDs in a human L-type CaV1.2 channel consisting of α1C and β3 subunits. We find that, without α2δ-1, the channel complex displays a right-shifted voltage dependence such that currents mainly develop at nonphysiological membrane potentials because of very weak VSD-pore interactions. The presence of α2δ-1 facilitates channel activation by increasing the voltage sensitivity (i.e., the effective charge) of VSDs I-III. Moreover, the α2δ-1 subunit also makes VSDs I-III more efficient at opening the channel by increasing the coupling energy between VSDs II and III and the pore, thus allowing Ca influx within the range of physiological membrane potentials.


Activity Begins in Childhood (ABC) - inspiring healthy active behaviour in preschoolers: study protocol for a cluster randomized controlled trial.
Today's children are more overweight than previous generations and physical inactivity is a contributing factor. Modelling and promoting positive behaviour in the early years is imperative for the development of lifelong health habits. The social and physical environments where children spend their time have a powerful influence on behaviour. Since the majority of preschool children spend time in care outside of the home, this provides an ideal setting to examine the ability of an intervention to enhance movement skills and modify physical activity behaviour. This study aims to evaluate the efficacy of the Activity Begins in Childhood (ABC) intervention delivered in licensed daycare settings alone or in combination with a parent-driven home physical activity-promotion component to increase preschoolers' overall physical activity levels and, specifically, the time spent in moderate to vigorous physical activity. This study is a single site, three-arm, cluster-randomized controlled trial design with a daycare centre as the unit of measurement (clusters). All daycare centres in the National Capital region that serve children between the ages of 3 and 5, expressing an interest in receiving the ABC intervention will be invited to participate. Those who agree will be randomly assigned to one of three groups: i) ABC program delivered at a daycare centre only, ii) ABC program delivered at daycare with a home/parental education component, or iii) regular daycare curriculum. This study will recruit 18 daycare centres, 6 in each of the three groups. The intervention will last approximately 6 months, with baseline assessment prior to ABC implementation and follow-up assessments at 3 and 6 months. Physical activity is an acknowledged component of a healthy lifestyle and childhood experiences as it has an important impact on lifelong behaviour and health. Opportunities for physical activity and motor development in early childhood may, over the lifespan, influence the maintenance of a healthy body weight and reduce cardiovascular disease risk. If successful, the ABC program may be implemented in daycare centres as an effective way of increasing healthy activity behaviours of preschoolers. Current Controlled Trials: ISRCTN94022291. Registered in December 2012, first cluster randomized in April 2013.


Sealing Ability of MTA Used in Perforation Repair of Permanent Teeth; Literature Review.
There were several materials used to seal different types of perforation defects. MTA is one of these restorative materials that is considered the most effective, biocompatible, non-toxic, and non-irritant; promote bone healing and cementum regeneration. The objective of this article was to review and summarize the sealing ability of MTA compared with the other materials used for sealing different types of root perforations of permanent teeth. A literature search was conducted using Medline, accessed via the National Library of Medicine Pub Med from 2005 to 2015 searching for articles related to sealing ability of MTA. This study found that factors affecting prognosis are the size, site of the perforation and time elapsed as well as the repair material. MTA is an important filling material to be used for sealing different types of perforations when perforated sites sealed immediately with MTA.


EIDOS: a mechanistic classification of adverse drug effects.
The mechanisms of adverse drug effects have not been adequately classified. Here, we propose a comprehensive mechanistic classification of adverse drug effects that considers five elements: the Extrinsic chemical species (E) that initiates the effect; the Intrinsic chemical species (I) that it affects; the Distribution (D) of these species in the body; the (physiological or pathological) Outcome (O); and the Sequela (S), which is the adverse effect. This classification, which we have called EIDOS, describes the mechanism by which an adverse effect occurs; it complements the DoTS classification of adverse effects (based on clinical pharmacology), which takes into account Dose responsiveness, Time course, and Susceptibility factors. Together, these two classification systems, mechanistic and clinical, comprehensively delineate all the important aspects of adverse drug reactions; they should contribute to areas such as drug development and regulation, pharmacovigilance, monitoring therapy, and the prevention, diagnosis, and treatment of adverse drug effects.


The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels.
Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). Here we investigated whether the composition of the N-glycan affected Kv1.4 cell surface expression. Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. This N-glycan had the most negative effect on cell surface expression of Kv1.4. It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4.


Sliding clamp of the bacteriophage T4 polymerase has open and closed subunit interfaces in solution.
The sliding clamps of bacteriophage T4 (gp45), Escherichia coli (beta clamp), and yeast (PCNA) are required for processive DNA synthesis by their cognate DNA polymerases. The X-ray crystal structures of all three of these clamps have been shown to be closed, circular complexes. This paper reports investigations of the solution structure of bacteriophage T4 gp45 by analytical ultracentrifugation, fluorescence, and hydrodynamic modeling. Mutants of gp45 with inter- and intrasubunit disulfide bonds were created to alter the solution structure of gp45, with additional mutagenesis used to investigate the importance of the proline-rich loop region found between the two domains of each gp45 monomer. The wild-type gp45 trimer assembles from monomers cooperatively with a dissociation constant of 0.21 microM2 and values between 0.088 and 0. 32 microM2 for the mutants. Velocity ultracentrifugation experiments showed that wild-type gp45 possesses a sedimentation coefficient strongly dependent on concentration, typical of asymmetric or elongated molecules, that when extrapolated to zero concentration yields a sedimentation coefficient of 4.0 S. The loop and the disulfide mutants exhibited sedimentation coefficients with little concentration dependence, typical of symmetric or spherical molecules, that when extrapolated to zero concentration yielded sedimentation coefficients of 4.4-4.8 S. The lower sedimentation coefficient in the former case is consistent with wild-type gp45 being more asymmetric or elongated than the mutant forms. Fluorescence-resonance energy-transfer experiments were used to measure the distance between two amino acids (W91 and V162C-coumarin) on opposite sides of the gp45 subunit interface. For an intrasubunit disulfide mutant, the distance between these two amino acids was determined to be 19 A (14 A in the X-ray crystal structure), consistent with a closed complex. For the mutants without intrasubunit disulfides, the efficiency of fluorescence-resonance energy transfer was in accord with a model of gp45 being an open complex composed of two closed subunit interfaces and a third open interface separated by a distance of 35-38 A. The collective data supplemented with hydrodynamic modeling were consistent with gp45 subunit separation achieved within the plane of the gp45 ring.


Amelioration of Uremic Toxin Indoxyl Sulfate-Induced Osteoblastic Calcification by SET Domain Containing Lysine Methyltransferase 7/9 Protein.
Vascular calcification (VC) is a very common phenomenon in patients with chronic kidney disease (CKD). It has been reported that some histone methylation play a role in VC as an epigenetic regulator. Indoxyl sulfate (IS) is a protein-bound uremic toxin that has been proven as one of the major risk factors of cardiovascular disease in CKD. SET domain containing lysine methyltransferase 7/9 (SET7/9) is one of the important histone methyltransferases. This study aimed to determine the effect of IS on the expression of SET7/9 and the role of SET7/9 in IS-induced osteoblastic differentiation and calcification of vascular smooth muscle cells (VSMCs). VSMCs were incubated with various concentrations of IS for different durations to assess osteoblastic differentiation and expression of SET7/9. Western blot analysis and quantitative real-time polymerase chain reaction were performed to assess the protein and mRNA levels of SET7/9 respectively. The calcium content was measured to evaluate calcification. Osteoblastic differentiation and calcification of VSMCs and downregulation of the expression of SET7/9 were observed after IS treatment. The autophagy was activated after treatment with IS, whereas the inhibition of the autophagy partially attenuated the effect of IS on both the stimulation of the expression of runt-related transcription factor 2 and calcium deposition. Our data demonstrated that SET7/9 downregulation and autophagy activation may be the key mechanism of IS-induced VC in CKD.


Quality of Health Management Information System for Maternal & Child Health Care in Haryana State, India.
Despite increasing importance being laid on use of routine data for decision making in India, it has frequently been reported to be riddled with problems. Evidence suggests lack of quality in the health management information system (HMIS), however there is no robust analysis to assess the extent of its inaccuracy. We aim to bridge this gap in evidence by assessing the extent of completeness and quality of HMIS in Haryana state of India. Data on utilization of key maternal and child health (MCH) services were collected using a cross-sectional household survey from 4807 women in 209 Sub-Centre (SC) areas across all 21 districts of Haryana state. Information for same services was also recorded from HMIS records maintained by auxiliary nurse midwives (ANMs) at SCs to check under- or over-recording (Level 1 discordance). Data on utilisation of MCH services from SC ANM records, for a subset of the total women covered in the household survey, were also collected and compared with monthly reports submitted by ANMs to assess over-reporting while report preparation (Level 2 discordance) to paint the complete picture for quality and completeness of routine HMIS. Completeness of ANM records for various MCH services ranged from 73% for DPT1 vaccination dates to 94.6% for dates of delivery. Average completeness level for information recorded in HMIS was 88.5%. Extent of Level 1 discordance for iron-folic acid (IFA) supplementation, 3 or more ante-natal care (ANC) visits and 2 Tetanus toxoid (TT) injections was 41%, 16% and 2% respectively. In 48.2% cases, respondents from community as well as HMIS records reported at least one post-natal care (PNC) home visit by ANM. Extent of Level 2 discordance ranged from 1.6% to 6%. These figures were highest for number of women who completed IFA supplementation, contraceptive intra-uterine device insertion and provision of 2nd TT injection during ANC. HMIS records for MCH services at sub-centre level in Haryana state were satisfactory in terms of completeness. However, there were significant differences in terms of reported and evaluated coverage of MCH services. Quality of HMIS needs to be improved in order to make it relevant for public health program planning and research.


Contrast class cues and performance facilitation in a hypothesis-testing task: evidence for an iterative counterfactual model.
Hypothesis-testing performance on Wason's (Quarterly Journal of Experimental Psychology 12:129-140, 1960) 2-4-6 task is typically poor, with only around 20% of participants announcing the to-be-discovered "ascending numbers" rule on their first attempt. Enhanced solution rates can, however, readily be observed with dual-goal (DG) task variants requiring the discovery of two complementary rules, one labeled "DAX" (the standard "ascending numbers" rule) and the other labeled "MED" ("any other number triples"). Two DG experiments are reported in which we manipulated the usefulness of a presented MED exemplar, where usefulness denotes cues that can establish a helpful "contrast class" that can stand in opposition to the presented 2-4-6 DAX exemplar. The usefulness of MED exemplars had a striking facilitatory effect on DAX rule discovery, which supports the importance of contrast-class information in hypothesis testing. A third experiment ruled out the possibility that the useful MED triple seeded the correct rule from the outset and obviated any need for hypothesis testing. We propose that an extension of Oaksford and Chater's (European Journal of Cognitive Psychology 6:149-169, 1994) iterative counterfactual model can neatly capture the mechanisms by which DG facilitation arises.


A hierarchical Bayesian approach to adaptive vision testing: A case study with the contrast sensitivity function.
Measurement efficiency is of concern when a large number of observations are required to obtain reliable estimates for parametric models of vision. The standard entropy-based Bayesian adaptive testing procedures addressed the issue by selecting the most informative stimulus in sequential experimental trials. Noninformative, diffuse priors were commonly used in those tests. Hierarchical adaptive design optimization (HADO; Kim, Pitt, Lu, Steyvers, & Myung, 2014) further improves the efficiency of the standard Bayesian adaptive testing procedures by constructing an informative prior using data from observers who have already participated in the experiment. The present study represents an empirical validation of HADO in estimating the human contrast sensitivity function. The results show that HADO significantly improves the accuracy and precision of parameter estimates, and therefore requires many fewer observations to obtain reliable inference about contrast sensitivity, compared to the method of quick contrast sensitivity function (Lesmes, Lu, Baek, & Albright, 2010), which uses the standard Bayesian procedure. The improvement with HADO was maintained even when the prior was constructed from heterogeneous populations or a relatively small number of observers. These results of this case study support the conclusion that HADO can be used in Bayesian adaptive testing by replacing noninformative, diffuse priors with statistically justified informative priors without introducing unwanted bias.


Proteomics analysis of human obesity reveals the epigenetic factor HDAC4 as a potential target for obesity.
Sedentary lifestyle and excessive energy intake are prominent contributors to obesity; a major risk factors for the development of insulin resistance, type 2 diabetes and cardiovascular diseases. Elucidating the molecular mechanisms underlying these chronic conditions is of relevant importance as it might lead to the identification of novel anti-obesity targets. The purpose of the current study is to investigate differentially expressed proteins between lean and obese subjects through a shot-gun quantitative proteomics approach using peripheral blood mononuclear cells (PBMCs) extracts as well as potential modulation of those proteins by physical exercise. Using this approach, a total of 47 proteins showed at least 1.5 fold change between lean and obese subjects. In obese, the proteomic profiling before and after 3 months of physical exercise showed differential expression of 38 proteins. Thrombospondin 1 (TSP1) was among the proteins that were upregulated in obese subjects and then decreased by physical exercise. Conversely, the histone deacetylase 4 (HDAC4) was downregulated in obese subjects and then induced by physical exercise. The proteomic data was further validated by qRT-PCR, Western blot and immunohistochemistry in both PBMCs and adipose tissue. We also showed that HDAC4 levels correlated positively with maximum oxygen consumption (VO2 Max) but negatively with body mass index, percent body fat, and the inflammatory chemokine RANTES. In functional assays, our data indicated that ectopic expression of HDAC4 significantly impaired TNF-α-dependent activation of NF-κB, establishing thus a link between HDAC4 and regulation of the immune system. Together, the expression pattern of HDAC4 in obese subjects before and after physical exercise, its correlation with various physical, clinical and metabolic parameters along with its inhibitory effect on NF-κB are suggestive of a protective role of HDAC4 against obesity. HDAC4 could therefore represent a potential therapeutic target for the control and management of obesity and presumably insulin resistance.


Targeted anticytokine therapy in patients with chronic heart failure: results of the Randomized Etanercept Worldwide Evaluation (RENEWAL).
Studies in experimental models and preliminary clinical experience suggested a possible therapeutic role for the soluble tumor necrosis factor antagonist etanercept in heart failure. Patients with New York Heart Association class II to IV chronic heart failure and a left ventricular ejection fraction < or =0.30 were enrolled in 2 clinical trials that differed only in the doses of etanercept used. In RECOVER, patients received placebo (n=373) or subcutaneous etanercept in doses of 25 mg every week (n=375) or 25 mg twice per week (n=375). In RENAISSANCE, patients received placebo (n=309), etanercept 25 mg twice per week (n=308), or etanercept 25 mg 3 times per week (n=308). The primary end point of each individual trial was clinical status at 24 weeks. Analysis of the effect of the 2 higher doses of etanercept on the combined outcome of death or hospitalization due to chronic heart failure from the 2 studies was also planned (RENEWAL). On the basis of prespecified stopping rules, both trials were terminated prematurely owing to lack of benefit. Etanercept had no effect on clinical status in RENAISSANCE (P=0.17) or RECOVER (P=0.34) and had no effect on the death or chronic heart failure hospitalization end point in RENEWAL (etanercept to placebo relative risk=1.1, 95% CI 0.91 to 1.33, P=0.33). The results of RENEWAL rule out a clinically relevant benefit of etanercept on the rate of death or hospitalization due to chronic heart failure.


Cortex-wide BOLD fMRI activity reflects locally-recorded slow oscillation-associated calcium waves.
Spontaneous slow oscillation-associated slow wave activity represents an internally generated state which is characterized by alternations of network quiescence and stereotypical episodes of neuronal activity - slow wave events. However, it remains unclear which macroscopic signal is related to these active periods of the slow wave rhythm. We used optic fiber-based calcium recordings of local neural populations in cortex and thalamus to detect neurophysiologically defined slow calcium waves in isoflurane anesthetized rats. The individual slow wave events were used for an event-related analysis of simultaneously acquired whole-brain BOLD fMRI. We identified BOLD responses directly related to onsets of slow calcium waves, revealing a cortex-wide BOLD correlate: the entire cortex was engaged in this specific type of slow wave activity. These findings demonstrate a direct relation of defined neurophysiological events to a specific BOLD activity pattern and were confirmed for ongoing slow wave activity by independent component and seed-based analyses.


Intracellular trafficking in Drosophila visual system development: a basis for pattern formation through simple mechanisms.
Intracellular trafficking underlies cellular functions ranging from membrane remodeling to receptor activation. During multicellular organ development, these basic cell biological functions are required as both passive machinery and active signaling regulators. Exocytosis, endocytosis, and recycling of several key signaling receptors have long been known to actively regulate morphogenesis and pattern formation during Drosophila eye development. Hence, intracellular membrane trafficking not only sets the cell biological stage for receptor-mediated signaling but also actively controls signaling through spatiotemporally regulated receptor localization. In contrast to eye development, the role of intracellular trafficking for the establishment of the eye-to-brain connectivity map has only recently received more attention. It is still poorly understood how guidance receptors are spatiotemporally regulated to serve as meaningful synapse formation signals. Yet, the Drosophila visual system provides some of the most striking examples for the regulatory role of intracellular trafficking during multicellular organ development. In this review we will first highlight the experimental and conceptual advances that motivate the study of intracellular trafficking during Drosophila visual system development. We will then illuminate the development of the eye, the eye-to-brain connectivity map and the optic lobe from the perspective of cell biological dynamics. Finally, we provide a conceptual framework that seeks to explain how the interplay of simple genetically encoded intracellular trafficking events governs the seemingly complex cellular behaviors, which in turn determine the developmental product.


Thrombolysis of occluded infra-inguinal bypass grafts: is it worthwhile?
Thrombosis of an infra-inguinal bypass graft often results in a limb-threatening ischemia. There is no consensus on the optimal management strategy. To analyse safety and efficacy of catheter-guided intra-arterial thrombolysis to re-open thrombosed infra-inguinal bypass grafts and to identify factors that influence graft patency and limb salvage rate after thrombolytic procedures. A continuous cohort-study of 106 thrombolytic procedures between 1993 to 2008. Despite a high initial success rate (76%), the mid-term results are less convincing, with a 58% re-occlusion rate at 45 months follow-up. Thrombosed vein grafts, old (2 weeks or more) occlusions, poor run-off and failure to identify or rectify an underlying causative stenosis are determinant for a poor long-term outcome of thrombolytic procedures. The outcome results of author's experience are consistent with literature reports. Thrombolysis of occluded infra-inguinal bypass grafts should be limited to selective cases (recent occlusion, prosthetic or vein graft in place since 1 year or more, critical limb ischemia). Despite its obvious advantages, the long-term outcome of thrombolytic procedures is deceiving. The inherent risk of hemorrhagic complications should also be taken in account at the decision making of treatment strategy. The question whether, in general, catheter-guided selective intra-arterial thrombolysis offers a significant advantage over operative revascularisation (thrombectomy or new bypass) remains unanswered. A more selective approach seems to favour thrombolysis as most appropriate strategy in the management of recent (< 2 weeks) thromboses of grafts in place since at least 1 year.


Urinary Viral Shedding of COVID-19 and its Clinical Associations: A Systematic Review and Meta-analysis of Observational Studies.
To review the current literature on the presence of COVID-19 virus in the urine of infected patients and to explore the clinical features that can predict the presence of COVID-19 in urine. A systematic review of published literature between 30th December 2019 and 21st June 2020 was conducted on Pubmed, Google Scholar, Ovid, Scopus, and ISI web of science. Studies investigating urinary viral shedding of COVID-19 in infected patients were included. Two reviewers selected relative studies and performed quality assessment of individual studies. Meta-analysis was performed on the pooled case reports and cohort with a sample size of ≥ 9. Thirty-nine studies were finally included in the systematic review; 12 case reports, 26 case series, and one cohort study. Urinary samples from 533 patients were investigated. Fourteen studies reported the presence of COVID-19 in the urinary samples from 24 patients. The crude overall rate of COVID-19 detection in urinary samples was 4.5%. Considering case series and cohorts with a sample size of ≥ 9, the estimated viral shedding frequency was 1.18 % (CI 95%: 0.14 - 2.87) in the meta-analysis. Urinary viral load in most reports were lower than rectal or oropharyngeal samples. In adult patients, urinary shedding of COVID-19 was commonly detected in patients with moderate to severe disease (16 adult patients with moderate or severe disease versus two adult patients with mild disease). In children, urinary viral shedding of COVID-19 was reported in 4 children who all suffered from mild disease. Urinary viral shedding of COVID-19 was detected from day 1 to day 52 after disease onset. The pathogenicity of virus isolated from urine has been demonstrated in cell culture media in one study while another study failed to reveal replication of isolated viral RNA in cell cultures. Urinary symptoms were not attributed to urinary viral shedding. While COVID-19 is rarely detected in urine of infected individuals, infection transmission through urine still remains possible. In adult patients, infected urine is more likely in the presence of moderate or severe disease. Therefore, caution should be exerted when dealing with COVID-19 infected patients during medical interventions like endoscopy and urethral catheterization especially in symptomatic adult patients while in children caution should be exerted regardless of symptoms.


Ca2+ -dependent inactivation of the mitochondrial Ca2+ uniporter involves proton flux through the ATP synthase.
Stimulation of receptors on the surface of animal cells often evokes cellular responses by raising intracellular Ca(2+) concentration. The rise in cytoplasmic Ca(2+) drives a plethora of processes, including neurotransmitter release, muscle contraction, and cell growth and proliferation. Mitochondria help shape intracellular Ca(2+) signals through their ability to rapidly take up significant amounts of Ca(2+) from the cytosol via the uniporter, a Ca(2+)-selective ion channel in the inner mitochondrial membrane. The uniporter is subject to inactivation, whereby a sustained cytoplasmic Ca(2+) rise prevents further Ca(2+) uptake. In spite of its importance in intracellular Ca(2+) signaling, little is known about the mechanism underlying uniporter inactivation. Here, we report that maneuvers that promote matrix alkalinisation significantly reduce inactivation whereas acidification exacerbates it. We further show that the F(1)F(0)-ATP synthase complex is an important source of protons for inactivation of the uniporter. These findings identify a novel molecular mechanism that regulates the activity of this ubiquitous intracellular Ca(2+) channel, with implications for intracellular Ca(2+) signaling and aerobic ATP production.


Dosimetric aspects of inverse-planned modulated-arc total-body irradiation.
To develop optimal beam parameters and to verify the dosimetric aspects of the recently developed modulated-arc total-body irradiation (MATBI) technique, which delivers an inverse-planned dose to the entire body using gantry rotation. The patient is positioned prone and supine underneath the gantry at about 2 m source-to-surface distance (SSD). Then, up to 28 beams irradiate the patient from different gantry angles. Based on full-body computed-tomography (CT) images of the patient, the weight of each beam is optimized, using inverse planning, to create a uniform body dose. This study investigates how to best simulate patients and the ideal beam setup parameters, such as field size, number of beams, and beam geometry, for treatment time and dose homogeneity. In addition, three anthropomorphic water phantoms were constructed and utilized to verify the accuracy of dose delivery, with both diode array and ion chamber measurements. Furthermore, to improve the accuracy of the new technique, a beam model is created specifically for the extended-SSD positioning for MATBI. Low dose CT scans can be utilized for dose calculations without affecting the accuracy. The largest field size of 40 × 40 cm(2) was found to deliver the most uniform dose in the least amount of time. Moreover, a higher number of beams improves dose homogeneity. The average dose discrepancy between ion chamber measurements and extended-SSD beam model calculations was 1.2%, with the largest discrepancy being 3.2%. This average dose discrepancy was 1.4% with the standard beam model for delivery at isocenter. The optimum beam setup parameters, regarding dose uniformity and treatment duration, are laid out for modulated-arc TBI. In addition, the presented dose measurements show that these treatments can be delivered accurately. These measurements also indicated that a new beam model did not significantly improve the accuracy of dose calculations. The optimum beam setup parameters along with the measurements performed to ensure accurate dose delivery serve as a useful guide for the clinical implementation of MATBI.


A battery approach for measuring neuropsychological change.
The accurate interpretation of serial neuropsychological testing is a fundamental activity of neuropsychologists in clinical and research settings. Although prior research supports the construct validity of several individual tests for the reliable measurement of cognitive change, neuropsychologists are often interested in detecting changes in cognitive functioning across a battery of tests. In the present study, we examined the specificity of a modified Reliable Change Index (RCI) methodology applied across a focused battery of commonly used neuropsychological tests. Fifty-seven healthy controls underwent neuropsychological assessment at two time points separated by approximately 1 year. Test-retest reliability coefficients and standard RCI confidence intervals for the individual tests were broadly comparable with prior research in healthy populations. Battery change scores were generated by calculating z-scores of change for each individual test, which were summed across the entire test battery. The RCI methodology was applied to the summed z-score to provide a 90% confidence interval as an indicator of overall cognitive stability. These battery RCI normative standards demonstrated adequate specificity when applied to 29 persons with HIV-1 infection who were classified as medically and neurologically stable. Findings from this study may be useful for both clinicians and researchers seeking normative standards for determining reliable changes in performance across a commonly used battery of neuropsychological tests.


MOET Efficiency in a Spanish Herd of Japanese Black Heifers and Analysis of Environmental and Metabolic Determinants.
Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016-2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production (p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling (p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; (p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea (p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients' metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy.


The therapeutic potential of mesenchymal stem cell transplantation as a treatment for multiple sclerosis: consensus report of the International MSCT Study Group.
Current therapies for multiple sclerosis effectively reduce inflammation, but do little in terms of repair to the damaged central nervous system. Cell-based therapies may provide a new strategy for bolstering regeneration and repair through neuro-axonal protection or remyelination. Mesenchymal stem cells modulate pathological responses in experimental autoimmune encephalitis, alleviating disease, but also stimulate repair of the central nervous system through the release of soluble factors. Autologous and allogeneic mesenchymal stem cells have been safely administered to individuals with hemato-oncological diseases and in a limited number of patients with multiple sclerosis. It is therefore reasonable to move mesenchymal stem cells transplantation into properly controlled human studies to explore their potential as a treatment for multiple sclerosis. Since it is likely that the first such studies will probably involve only small numbers of patients in a few centers, we formed an international panel comprising multiple sclerosis neurology and stem cell experts, as well as immunologists. The aims were to derive a consensus on the utilization of mesenchymal stem cells for the treatment of multiple sclerosis, along with protocols for the culture of the cells and the treatment of patients. This article reviews the consensus derived from our group on the rationale for mesenchymal stem cell transplantation, the methodology for generating mesenchymal stem cells and the first treatment protocol for multiple sclerosis patients.


'A giant mess'--making sense of complexity in the accounts of people with fibromyalgia.
The diagnosis of fibromyalgia is based on self-report and indirect measures and thus is unavoidably influenced by patients' own understanding of their symptoms. In order to provide appropriate support for people with fibromyalgia, it is important to understand variation in patients' interpretations of their own symptoms. Twenty people with fibromyalgia participated in email interviews exploring their experiences, history and diagnosis. Respondents answered a series of questions in their own time. Rich accounts were elicited. A hermeneutic phenomenological approach linked two stages of analysis. In the first instance, an in-depth, inductive analysis was developed around a subset of eight transcripts, using interpretative phenomenological analysis. The outcomes of this work were then used to inform a template analysis, which was applied to the remaining 12 transcripts, in order to extend and check the credibility of the in-depth analysis. Participants described enduring the course of a 'giant mess' of unpleasant symptoms, some of which were understood to be symptoms of fibromyalgia and some the interactive or parallel effects of comorbid illness. The respondents also demonstrated their considerable efforts at imposing order and sense on complexity and multiplicity, in terms of the instability of their symptoms. They expressed ambivalence towards diagnosis, doctors and medication, and we noted that each of the above areas appeared to come together to create a context of relational uncertainty, which undermined the security of connections to family, friends, colleagues and the workplace. Three key issues were discussed. First, there was not one overall symptom (e.g., pain) driving the unpleasantness of fibromyalgia; second, participants spent excessive time and energy trying to manage forces outside their control; third, because there is no definitive 'fibromyalgia experience', each diagnosis is unique, and our participants often appeared to be struggling to understand the course of their illness. Issues of stigma and legitimacy need to be considered carefully by health professionals in the context of the complex and uncertain experience of patients.


Management of penetrating spinal cord injuries in a non spinal centre: experience at Enugu, Nigeria.
Penetrating injuries of the spinal cord are among the most dangerous of injuries. They are often associated with injuries to other vital organs of the body, which may demand priority attention. The objectives of this study were to determine the pattern of the penetrating spinal cord injuries and to evaluate outcome of treatment in our setting. This is a retrospective study of all penetrating spinal cord injuries seen at the National Orthopaedic Hospital, Enugu over a fifteen-year period (April 1990 March 2005). Information about the demographics aetiology, level of injury, associated injuries, time of presentation, duration of hospitalization and outcome of treatment were retrieved from patients case notes. Data were analyzed using SPSS for windows version 11. There were 22 patients, 13 males and 9 females, giving a M:F ratio of 1.7:1. Gunshot injury was the most common aetiological factor. The thoracic spine {9 (41%)} was most often involved. Chest and abdominal injuries were common associated injuries in 5 cases. The circumstances of the injury were mostly armed robbery attack {13 (59%)}. On admission the neurological status was Frankel grade A in 20(91%) cases. Period of hospitalization ranged from 1 week to 36 weeks with a mean of 11 weeks. Pressure sore was the most common complication that delayed rehabilitation. Five (23%) patients with injury at cervical level died from respiratory failure. Penetrating spinal cord injuries are relatively rare and demand extra care. Early recognition of associated injuries, minimal wound excision and antibiotic therapy give good result.