Maternal pre-pregnancy BMI and reproductive health in adult sons: a study in the Danish National Birth Cohort.
Is maternal pre-pregnancy BMI associated with semen quality, testes volume, and reproductive hormone levels in sons? Maternal pre-pregnancy BMI was associated with an altered reproductive hormone profile in young adult sons, characterized by higher levels of oestradiol, LH, and free androgen index (FAI) and lower levels of sex hormone-binding globulin (SHBG) in sons born of mothers with pre-pregnancy overweight and obesity. Evidence suggests that maternal pre-pregnancy BMI may influence reproductive health later in life. Only one pilot study has investigated the association between maternal pre-pregnancy BMI and reproductive health outcomes in sons, suggesting that a high BMI was associated with impaired reproductive function in the adult sons. A population-based follow-up study of 1058 young men from the Fetal Programming of Semen Quality (FEPOS) cohort nested within the Danish National Birth Cohort (DNBC), 1998-2019, was carried out. In total, 1058 adult sons (median age 19 years, 2 months), born 1998-2000 by mothers included in the DNBC, participated in FEPOS. At a clinical examination, they provided a semen and blood sample, measured their testes volume, and had height and weight measured. Maternal pre-pregnancy BMI was obtained by self-report in early pregnancy. Semen characteristics, testes volume, and reproductive hormone levels were analysed according to maternal pre-pregnancy BMI categories and as restricted cubic splines using negative binomial and ordinary least square regression models. Mediation analyses examined potential mediation by the sons' birthweight, pubertal timing, fat mass, and BMI. Additional analyses investigated the role of paternal BMI in the potential associations between maternal BMI and reproductive health outcomes. We found no consistent associations between maternal pre-pregnancy BMI and semen characteristics or testes volume. Sons of mothers with higher pre-pregnancy BMI had higher oestradiol and lower SHBG levels, both in a dose-dependent manner. Sons of mothers with pre-pregnancy obesity (≥30 kg/m2) had higher LH levels and a higher FAI than sons born by mothers with normal pre-pregnancy BMI (18.5-24.9 kg/m2). The mediation analyses suggested that the effect of maternal pre-pregnancy BMI on higher levels of oestrogen, LH, and FAI was partly mediated by the sons' birthweight, in addition to adult fat mass and BMI measured at the clinical examination, whereas most of the effect on lower levels of SHBG was primarily mediated by the sons' own fat mass and BMI. Paternal BMI was not a strong confounder of the associations in this study. This study was based in a population-based cohort with a low prevalence of overweight and obesity in both mothers and adult sons. Some men (10%) had blood for reproductive hormone assessment drawn in the evening. While several potential confounding factors were accounted for, this study's inherent risk of residual and unmeasured confounding precludes provision of causal estimates. Therefore, caution should be given when interpreting the causal effect of maternal BMI on sons' reproductive health. Given the widespread occurrence of overweight and obesity among pregnant women, it is imperative to thoroughly examine the potential consequences for reproductive hormone levels in adult sons. The potential effects of maternal pre-pregnancy obesity on sons' reproductive hormone profile may potentially be partly avoided by the prevention of overweight and obesity in the sons. The project was funded by the Lundbeck Foundation (R170-2014-855), the Capital Region of Denmark, Medical doctor Sofus Carl Emil Friis and spouse Olga Doris Friis's Grant, Axel Muusfeldt's Foundation (2016-491), AP Møller Foundation (16-37), the Health Foundation, Dagmar Marshall's Fond, Aarhus University, Independent Research Fund Denmark (9039-00128B), and the European Union (ERC, BIOSFER, 101071773). Views and opinions expressed are, however, those of the authors only and do not necessarily reflect those of the European Union or the European Research Council. Neither the European Union nor the granting authority can be held responsible. The authors declare that they have no conflict of interest. N/A.


Ventral-inlay buccal mucosal graft urethroplasty for female urethral stricture.
The aim of the study is to present our initial experience with ventral-inlay buccal mucosal graft urethroplasty (VI-BMGU) in female urethral stricture disease (USD). Between May 2016 and June 2018, 12 women with USD underwent VI-BMGU. All women were evaluated preoperatively with the American Urological Association (AUA) symptom score, uroflowmetry, calibration with a 12 Fr catheter, and ultrasonography with postvoid residual (PVR) urine measurement. Intraoperative confirmation of stricture was done with a 6 Fr cystoscope. Postoperatively, the women were followed at 3, 6, and 12 months after surgery with AUA symptom score, uroflowmetry, and PVR estimation. Increase in AUA symptom score, maximum flow rate (Qmax) <12 ml/s, and failure to calibrate with 18 Fr catheters were considered as indicative of recurrence of the disease. The mean age of the patients was 41 years. The mean follow-up period was 18 months. All women voided successfully after catheter removal. There was an improvement in AUA symptom score and Qmax and a reduction in PVR at 3, 6, and 12 months. One woman had recurrence of stricture at 6 months and was treated by urethral dilatation followed by the institution of a self-dilatation regimen. The success rate was 92% in our case series. VI-BMGU is a simple and safe method of urethroplasty in women. Studies with a larger sample size and a longer follow-up are required to document the long-term success of this procedure.


A chronic thromboembolic pulmonary hypertension catch-22 situation: inferior vena caval filter plays a pivotal role in an unlikely situation.
Chronic thromboembolic pulmonary hypertension (CTEPH) is a rare clinical entity that occurs in a small subset of acute pulmonary embolism (PE) cases and is surgically cured by pulmonary endarterectomy. We report a case of a 44-year-old female with a complex history of CTEPH treated by thromboendarterectomy who presented with a subdural hematoma while on warfarin. The patient eventually recovered by a multidisciplinary approach, use of inferior vena caval filter, and effective anticoagulation management.


From current status to optimization of HCV treatment: Recommendations from an expert panel.
Chronic hepatitis C virus (HCV) infection is a major public health problem at a global level, causing an enormous burden of hepatic and extra-hepatic morbidity and mortality. Treatment of chronic HCV (CHC) has been revolutionized in the last few years by the introduction of highly effective and well tolerated direct acting antiviral agents (DAAs) able to achieve >90% rates of sustained virological response (SVR) in many groups of patients, including those previously excluded from interferon-based regimens. For such reason interferon-free regimens are now the treatments of choice for all patients. Successful anti-HCV treatment can stop liver disease progression and can solve the HCV-related extra hepatic manifestations, eventually reducing both liver-related and overall mortality. Together with the rapidly accumulating data about the evolution of treatment landscape, different guidelines from national and international Liver Scientific Societies have been published until today. However, these recommendations may not be applied worldwide as, due to high treatment costs, most of them identify as priority groups only patients with advanced liver disease. Moreover some types of patients pose clinical management problems for which even the guidelines do not always provide useful answers. With the aim of treatment optimization by filling some of the gaps of the current guidelines and addressing the remaining unmet needs in practice, a group of Italian experts, experienced on treatment of HCV infection, met in Stresa in February 2016. The summary of all the considerations arising from this two-day meeting and the final statements are reported in this position paper.


Establishment and characterization of a cancer cell line derived from an aggressive childhood liver tumor.
Hepatoblastoma is a rare malignancy of childhood. The scarcity of adequate cell models has limited our understanding of this tumor. Here we describe and characterize a new human liver tumor cell line, Hep293TT, derived from an aggressive childhood hepatoblastoma. Hep293TT cells were established using primary tumor tissues from a 5-year-old Caucasian female child. This cell line has been maintained for more than 34 months and over 20 subcultures, and was characterized by histopathology, ELISA, genotype, cytogenetics, CGH array, immunohistochemistry, and molecular sequence analyses. Cells were confirmed to originate from parental tumor cells, secrete alpha-fetoprotein, and express hepatic markers and beta-catenin. Hep293TT cells were able to form colonies in soft agar. Tumorigenicity was demonstrated by induction of solid tumors after subrenal capsule injection in immunodeficient mice. Hep293TT cells demonstrated a highly aneuploid karyotype, and a whole genome CGH analysis revealed chromosomal imbalances in every chromosome. Allelotype analysis demonstrated loss of alleles at distal 11p15.5 as is typical of embryonal tumors. Both Hep293TT cells and the primary tumor contain a deletion of 351 nucleotides in beta-catenin, as has been seen in other hepatoblastoma tumors. The cell line expressed beta-catenin protein in both full-length and partially deleted forms, and expressed NOTCH2 protein characteristic of hepatoblasts. No mutation was detected in the APC, MYH, MLH1, or MSH2 genes. This cell line, Hep293TT, is a valuable resource for the study of childhood liver cancer and may potentially provide a tool in the development of new agents.


Protocols for Studying How Thyroid Hormone Receptors Trigger Adaptive Thermogenesis in Mice.
The ability of thyroid hormone T3 to stimulate energy expenditure and regulate different aspects of whole body metabolism has been recognized for over a 100 years. Indeed in 1895 Adolf Magnus Levy was already describing the influence of the thyroid on setting the basal metabolic rate. Now it has been well characterized that the level of circulating T3 is correlated with energy expenditure both in humans and in rodent models (Mullur et al., Physiol Rev 94:355-382, 2014; Silva, Thyroid 5:481-492, 1995). Hypothyroidism and hyperthyroidism are respectively associated with low and high energy expenditure leading to high and low body mass index. Moreover, T3 also reduces the level of cholesterol in the serum, which is one of the key risk factors to develop atherosclerosis (Mullur et al., Physiol Rev 94:355-382, 2014; Hak et al., Ann Intern Med 132:270-278, 2000). Given the worldwide pandemic of obesity and associated metabolic disorders in the last two decades, the metabolic activities of T3 gained a renewed interest. However, one has to keep in mind that despite its desirable actions, T3 cannot be used as a pharmacological agent since it also triggers unacceptable effects including tachycardia, lean mass loss, and osteoporosis. To solve this conundrum, a recent intense effort has been dedicated to understand how T3 fulfills its different activities, looking for the target tissues and the specific T3 receptor (TR) involved. Indeed recently it was shown that injection of T3 in the brain could efficiently trigger thermogenesis (López et al., Nat Med 16:1001-1008, 2010). This questioned the classical paradigm in which most metabolic actions were the result of local T3 action in the different metabolic tissues. To tackle this problem, many new models of transgenic mice carrying selective Knock-out and Knock-in mutation of TR in specific metabolic tissues are currently generated using conditional mutant alleles of THRA (Quignodon et al., Mol Endocrinol 21:2350-2360, 2007) and THRB (Billon et al., Endocrinology 155:2735-2745, 2014). In parallel some new compounds that allow the targeting of T3 to selective metabolic tissues (Finan et al., Cell 167:843-857, 2016) have also been obtained.In this chapter we will provide and comment two different protocols that are useful to study the adaptive thermogenesis in response to two physiological stresses: cold exposure and high fat diet feeding. They can also be used to test the thermogenic activity of the new designed compounds.


Idarucizumab, a specific reversal agent for dabigatran: mode of action, pharmacokinetics and pharmacodynamics, and safety and efficacy in phase 1 subjects.
The direct oral anticoagulants (DOACs) provide a number of clinical advantages over vitamin K antagonists for the treatment of thromboembolism, including improved efficacy and safety, as well as no need for regular monitoring of anticoagulant effect. However, as with all anticoagulants, bleeding complications may occur, and anticoagulant reversal may be required in specific clinical situations, such as in patients experiencing spontaneous or traumatic bleeds, or in anticoagulated patients requiring emergency surgery or other invasive procedures. Therefore, several reversal agents for the DOACs are in development. This includes the specific reversal agent idarucizumab, which has been approved by the U.S. Food and Drug Administration and the European Medicines Agency for use in patients treated with dabigatran when urgent reversal of its anticoagulant effects is needed. Idarucizumab is a humanized monoclonal antibody fragment that binds with high affinity to free and thrombin-bound dabigatran, resulting in an almost irreversibly bound idarucizumab-dabigatran complex and thereby neutralizing dabigatran's anticoagulant activity. The reversal of the anticoagulant effects of dabigatran by idarucizumab has been demonstrated in animal bleeding models, in healthy volunteers with a range of ages and renal function, and in anticoagulated patients. In the phase 1 trials, at doses of 2 g or greater, idarucizumab resulted in immediate and complete reversal of the dabigatran anticoagulant effects and was well tolerated. In the absence of dabigatran, idarucizumab showed no effect on coagulation parameters or thrombin formation. These findings provide initial evidence that idarucizumab could provide a safe and effective means of reversing anticoagulant activity in patients treated with dabigatran in need of emergency surgery or in emergency bleeding situations.


A dataset of microscopic peripheral blood cell images for development of automatic recognition systems.
This article makes available a dataset that was used for the development of an automatic recognition system of peripheral blood cell images using convolutional neural networks [1]. The dataset contains a total of 17,092 images of individual normal cells, which were acquired using the analyzer CellaVision DM96 in the Core Laboratory at the Hospital Clinic of Barcelona. The dataset is organized in the following eight groups: neutrophils, eosinophils, basophils, lymphocytes, monocytes, immature granulocytes (promyelocytes, myelocytes, and metamyelocytes), erythroblasts and platelets or thrombocytes. The size of the images is 360 × 363 pixels, in format jpg, and they were annotated by expert clinical pathologists. The images were captured from individuals without infection, hematologic or oncologic disease and free of any pharmacologic treatment at the moment of blood collection. This high-quality labelled dataset may be used to train and test machine learning and deep learning models to recognize different types of normal peripheral blood cells. To our knowledge, this is the first publicly available set with large numbers of normal peripheral blood cells, so that it is expected to be a canonical dataset for model benchmarking.


Evaluation of dietary supplementation of a novel microbial muramidase on gastrointestinal functionality and growth performance in broiler chickens.
This study was conducted to assess the effect of dietary supplementation of Muramidase 007 to broiler chickens on gastrointestinal functionality, evaluating growth performance, apparent ileal digestibility, intestinal histomorphology, vitamin A in plasma and cecal microbiota. A total of 480 one-day male chicks (Ross 308) were distributed in 16 pens allocated in 2 experimental diets: the control diet (CTR) without feed enzymes, coccidiostat or growth promoters, and the experimental diet (MUR): CTR supplemented with 35,000 units (LSU(F))/kg of the Muramidase 007. Digesta and tissue samples were obtained on days 9 and 36 of the study. A lower feed conversion ratio was observed in the MUR treatment. Apparent ileal digestibility of DM, organic matter and energy were improved by Muramidase 007. It was also observed that MUR improved digestibility of total fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids, and content of vitamin A in plasma at day 9 (P < 0.05). Histomorphological analysis of jejunum samples revealed no differences in the villus height or crypt depth; but a higher number of goblet cells and intraepithelial lymphocytes at day 36 with MUR. No differences were observed in plate counts of enterobacteria or Lactobacillus along the gastrointestinal tract, neither on the cecal short-chain fatty acids. An statistical trend was observed for reduction of cecal clostridia at day 9 for MUR. Analysis of cecal microbiota structure by 16S rRNA gene sequencing revealed relevant changes correlated to age. At day 9, broilers receiving MUR showed decreased alpha diversity compared to CTR that was not detected at day 36. Changes in specific taxonomic groups with an increase in Lactobacillus genus were identified. In conclusion, evaluation of the variables in this study indicates that dietary Muramidase 007 contributes to improve feed conversation ratio and gastrointestinal function in broiler chickens. Effects could have been mediated by slight shifts observed in the intestinal microbiota. More studies are guaranteed to fully understand the mechanisms involved.


The time line of threat processing and vagal withdrawal in response to a self-threatening stressor in cognitive avoidant copers: evidence for vigilance-avoidance theory.
Using a spatial cueing paradigm with emotional and neutral facial expressions as cues, we examined early and late patterns of information processing in cognitive avoidant coping (CAV). Participants were required to detect a target that appeared either in the same location as the cue (valid) or in a different location (invalid). Cue-target onset asynchrony (CTOA) was manipulated to be short (250 ms) or long (750 ms). CAV was associated with early facilitation and faster disengagement from angry faces. No effects were found for happy or neutral faces. After completing the spatial cueing task, participants prepared and delivered a public speech and heart rate variability (HRV) was recorded. Disengagement from angry faces was related to a decrease in HRV in response to this task. Together, these data suggest that CAV is related to early engagement followed by disengagement from threat-related cues that might impact physiological stress responses.


SLC9A6 mutations cause X-linked mental retardation, microcephaly, epilepsy, and ataxia, a phenotype mimicking Angelman syndrome.
Linkage analysis and DNA sequencing in a family exhibiting an X-linked mental retardation (XLMR) syndrome, characterized by microcephaly, epilepsy, ataxia, and absent speech and resembling Angelman syndrome, identified a deletion in the SLC9A6 gene encoding the Na(+)/H(+) exchanger NHE6. Subsequently, other mutations were found in a male with mental retardation (MR) who had been investigated for Angelman syndrome and in two XLMR families with epilepsy and ataxia, including the family designated as having Christianson syndrome. Therefore, mutations in SLC9A6 cause X-linked mental retardation. Additionally, males with findings suggestive of unexplained Angelman syndrome should be considered as potential candidates for SLC9A6 mutations.


A practical drug discovery project at the undergraduate level.
In this article, we describe a practical drug discovery project for third-year undergraduates. No previous knowledge of medicinal chemistry is assumed. Initial lecture workshops cover the basic principles; then students, in teams, seek to improve the profile of a weakly potent, insoluble phosphatidylinositide 3-kinase delta (PI3Kδ) inhibitor (1) through compound array design, molecular modelling, screening data analysis and the synthesis of target compounds in the laboratory. The project benefits from significant industrial support, including lectures, student mentoring and consumables. The aim is to make the learning experience as close as possible to real-life industrial situations. In total, 48 target compounds were prepared, the best of which (5b, 5j, 6b and 6ap) improved the potency and aqueous solubility of the lead compound (1) by 100-1000 fold and ≥tenfold, respectively.


Impact of climate change on larch budmoth cyclic outbreaks.
Periodic outbreaks of the larch budmoth Zeiraphera diniana population (and the massive forest defoliation they engender) have been recorded in the Alps over the centuries and are known for their remarkable regularity. But these have been conspicuously absent since 1981. On the other hand, budmoth outbreaks have been historically unknown in the larches of the Carpathian Tatra mountains. To resolve this puzzle, we propose here a model which includes the influence of climate and explains both the 8-9 year periodicity in the budmoth cycle and the variations from this, as well as the absence of cycles. We successfully capture the observed trend of relative frequencies of outbreaks, reproducing the dominant periodicities seen. We contend that the apparent collapse of the cycle in 1981 is due to changing climatic conditions following a tipping point and propose the recurrence of the cycle with a changed periodicity of 40 years - the next outbreak could occur in 2021. Our model also predicts longer cycles.


Spectrophotometric determination of pH and carbonate ion concentrations in seawater: Choices, constraints and consequences.
Accurate and precise marine CO2 system measurements are important for marine carbon cycle research and investigations of ocean acidification. Seawater pH is important because it can be used to characterize a wide range of chemical and biogeochemical processes. Saturation states of calcium carbonate minerals, which are directly proportional to carbonate ion concentration ([CO32-]), influence biogenic calcification and rates of carbonate dissolution. Spectrophotometric pH and carbonate ion measurements can both benefit greatly from the high sensitivity, stability, consistency and processing speed made possible through automation. Spectrophotometric methods are well-suited for shipboard, underway and in situ deployments under harsh conditions. Spectrophotometric pH measurements typically have a reproducibility of 0.0004-0.001 for shipboard and laboratory measurements and 0.0014-0.004 for in situ measurements. Shipboard spectrophotometric measurements of [CO32-] are becoming common on research expeditions. This review highlights the development of methods and instrumentation for spectrophotometric pH and [CO32-] measurements, and discusses the pros and cons of current technology. A comprehensive summary of the analytical merits of different flow analysis instruments is given. Aspects of measurement protocols that bear on the quality of pH and [CO32-] measurements, such as indicator purification, sample pretreatment, etc., are also described. Based on three decades of experience with seawater analysis, this review includes method recommendations and perspectives directly applicable or potentially applicable to pH and [CO32-] analysis of seawater.


Performance Enhancement Assessment and Coaching in US Army Special Operations: Rapidly Enhancing Performance Through Targeted, Tailored Feedback.
Performance enhancement coaching poses significant benefits to individuals and organizations, such as improved job satisfaction and goal achievement. Given their training and experience in assessment and feedback, operational psychologists assigned to Special Operations units are uniquely positioned to provide performance enhancement coaching tailored to Operators and enablers. A preliminary program evaluation was conducted of the Performance Enhancement Assessment and Coaching (PEAC) Program. A sample of 32 Operators and enablers assigned to a US Army Special Operations Forces (ARSOF) unit voluntarily participated in the PEAC Program and completed one 90-minute coaching session. Following their coaching session, Soldiers provided qualitative and quantitative feedback on their coaching experience. Soldiers overwhelmingly agreed that the PEAC Program was worth their time and helpful towards achieving their goals. Results indicate the PEAC Program enhanced Soldiers' perceived self-awareness, self-efficacy, and job performance. Results also suggest performance enhancement coaching may improve pass rates on interpersonally demanding Special Operations courses. Performance enhancement coaching delivers considerable value for Special Operations personnel and their organizations in relatively minimal time. Operational psychologist coaches (OPCs) assigned to Special Operations units can leverage their assessment skills to provide targeted, tailored performance enhancement coaching and increase value to their organizations.


Thymidine incorporation in nucleoside transport-deficient lymphoma cells.
Nucleoside transport deficiency in mammalian cells is associated with an inability to transport most nucleosides, growth resistance to a spectrum of cytotoxic nucleosides, and a loss of binding sites for 4-nitrobenzylthioinosine (NBMPR), a potent inhibitor of nucleoside transport. The nucleoside transport-deficient S49 T lymphoma cell line, AE1, however, was almost as capable of incorporating thymidine into TTP as the wild type parent provided thymidine was administered at a sufficiently high concentration. Consequently, AE1 cells were just as sensitive as wild type cells to the toxicity of high thymidine concentrations. In contrast, AE1 cells were highly resistant to almost all other cytotoxic nucleosides including the thymidine analogs, 5-bromodeoxyuridine and 5-fluoro-2'-deoxyuridine 5'-monophosphate. Despite having demonstrable ability to accumulate TTP, AE1 cells were unable to grow on hypoxanthine-amethopterin-thymidine (HAT)-containing medium. This was due to their inability to accumulate sufficient TTP from the low concentrations of thymidine present in HAT medium. AE1 cells possessed an incomplete thymidine transport deficiency, the extent of which was concentration dependent. The residual capacity for thymidine transport present in AE1 cells was insensitive to inhibition by 4-nitrobenzylthioinosine and could account both for their inability to grow on HAT medium and their sensitivity to cytotoxic concentrations of thymidine. Another nucleoside transport-deficient cell line, FURD-80-3-6, was similar to the AE1 cell line in its growth phenotype and NBMPR-binding site deficiency but differed in its decreased growth sensitivity to thymidine. That nucleoside transport deficiencies may vary in their completeness for different nucleosides has significance for the mechanism by which a single transporter can recognize a wide variety of nucleosides.


Obstructive sleep apnoea and type 2 diabetes mellitus: a bidirectional association.
Obstructive sleep apnoea and type 2 diabetes are common medical disorders that have important clinical, epidemiological, and public health implications. Research done in the past two decades indicates that obstructive sleep apnoea, through the effects of intermittent hypoxaemia and sleep fragmentation, could contribute independently to the development of insulin resistance, glucose intolerance, and type 2 diabetes. Conversely, type 2 diabetes might increase predisposition to, or accelerate progression of, obstructive and central sleep apnoea, possibly through the development of peripheral neuropathy and abnormalities of ventilatory and upper airway neural control. Although more research is needed to clarify the mechanisms underlying the bidirectional association between the two disorders, their frequent coexistence should prompt all health-care professionals to embrace clinical practices that include screening of a patient presenting with one disorder for the other. Early identification of obstructive sleep apnoea in patients with metabolic dysfunction, including type 2 diabetes, and assessment for metabolic abnormalities in those with obstructive sleep apnoea could reduce cardiovascular disease risk and improve the quality of life of patients with these chronic diseases.


The mouse telomerase RNA 5"-end lies just upstream of the telomerase template sequence.
Telomerase is a ribonucleoprotein enzyme with an essential RNA component. Embedded within the telomerase RNA is a template sequence for telomere synthesis. We have characterized the structure of the 5' regions of the human and mouse telomerase-RNA genes, and have found a striking difference in the location of the template sequence: Whereas the 5'-end of the human telomerase RNA lies 45 nt from the telomerase-RNA template sequence, the 5'-end of the mouse telomerase RNA lies just 2 nt from the telomerase-RNA template sequence. Analysis of genomic sequences flanking the 5'-end of the human and mouse telomerase RNA-coding sequences reveals similar promoter-element arrangements typical of mRNA-type promoters: a TATA box-like element and an upstream region containing a consensus CCAAT box. This putative promoter structure contrasts with that of the ciliate telomerase-RNA genes whose structure resembles RNA polymerase III U6 small nuclear RNA (snRNA) promoters. These and other comparisons suggest that, during evolution, both the RNA-polymerase specificity of telomerase RNA-gene promoters and, more recently, the position of the template sequence in the telomerase RNA changed.


Clinical Application of Interferon-γ Release Assays for the Prevention of Tuberculosis in Countries with Low Incidence.
Despite global efforts to control tuberculosis (TB) the estimated number of people who developed TB worldwide increased to an all-time record of more than 10 million in 2015. The goal of the World Health Organization (WHO) to reduce the global incidence of TB to less than 100 cases per million by 2035, cannot be reached unless TB prevention is markedly improved. There is a need for an improved vaccine that better protects individuals who are exposed to Mycobacterium tuberculosis from infection and active disease compared to the current M. bovis Bacille Calmette Guérin (BCG) vaccine. In the absence of such a vaccine, prevention relies on infection control measures and preventive chemotherapy for people with latent infection with M. tuberculosis (LTBI), who have the highest risk of progression to active TB. During the past decade, interferon-γ release assays (IGRAs) have increasingly replaced the tuberculin skin test as screening tools for the diagnosis of LTBI in countries with a low incidence of TB. Despite recent WHO guidelines on the management of LTBI, the definition of groups at risk for TB remains controversial, and the role of IGRAs for TB prevention in low-incidence countries remains uncertain. We reviewed the scientific literature and provide recommendations for the use of IGRAs for LTBI diagnosis in low-incidence countries. These recommendations are based on the number of patients needing treatment in order to prevent one case of TB. As the positive predictive value of IGRAs for the development of TB is sub-optimal, research must focus on the identification of alternative biomarkers that offer better predictive ability in order to substantially reduce the number needing treatment while improving the prevention of TB and improving the effectiveness of targeted preventive chemotherapy.


A novel role for RecA under non-stress: promotion of swarming motility in Escherichia coli K-12.
Bacterial motility is a crucial factor in the colonization of natural environments. Escherichia coli has two flagella-driven motility types: swimming and swarming. Swimming motility consists of individual cell movement in liquid medium or soft semisolid agar, whereas swarming is a coordinated cellular behaviour leading to a collective movement on semisolid surfaces. It is known that swimming motility can be influenced by several types of environmental stress. In nature, environmentally induced DNA damage (e.g. UV irradiation) is one of the most common types of stress. One of the key proteins involved in the response to DNA damage is RecA, a multifunctional protein required for maintaining genome integrity and the generation of genetic variation. The ability of E. coli cells to develop swarming migration on semisolid surfaces was suppressed in the absence of RecA. However, swimming motility was not affected. The swarming defect of a DeltarecA strain was fully complemented by a plasmid-borne recA gene. Although the DeltarecA cells grown on semisolid surfaces exhibited flagellar production, they also presented impaired individual movement as well as a fully inactive collective swarming migration. Both the comparative analysis of gene expression profiles in wild-type and DeltarecA cells grown on a semisolid surface and the motility of lexA1 [Ind-] mutant cells demonstrated that the RecA effect on swarming does not require induction of the SOS response. By using a RecA-GFP fusion protein we were able to segregate the effect of RecA on swarming from its other functions. This protein fusion failed to regulate the induction of the SOS response, the recombinational DNA repair of UV-treated cells and the genetic recombination, however, it was efficient in rescuing the swarming motility defect of the DeltarecA mutant. The RecA-GFP protein retains a residual ssDNA-dependent ATPase activity but does not perform DNA strand exchange. The experimental evidence presented in this work supports a novel role for RecA: the promotion of swarming motility. The defective swarming migration of DeltarecA cells does not appear to be associated with defective flagellar production; rather, it seems to be associated with an abnormal flagellar propulsion function. Our results strongly suggest that the RecA effect on swarming motility does not require an extensive canonical RecA nucleofilament formation. RecA is the first reported cellular factor specifically affecting swarming but not swimming motility in E. coli. The integration of two apparently disconnected biologically important processes, such as the maintenance of genome integrity and motility in a unique protein, may have important evolutive consequences.


Credentialing features: a platform to benchmark and optimize untargeted metabolomic methods.
The aim of untargeted metabolomics is to profile as many metabolites as possible, yet a major challenge is comparing experimental method performance on the basis of metabolome coverage. To date, most published approaches have compared experimental methods by counting the total number of features detected. Due to artifactual interference, however, this number is highly variable and therefore is a poor metric for comparing metabolomic methods. Here we introduce an alternative approach to benchmarking metabolome coverage which relies on mixed Escherichia coli extracts from cells cultured in regular and (13)C-enriched media. After mass spectrometry-based metabolomic analysis of these extracts, we "credential" features arising from E. coli metabolites on the basis of isotope spacing and intensity. This credentialing platform enables us to accurately compare the number of nonartifactual features yielded by different experimental approaches. We highlight the value of our platform by reoptimizing a published untargeted metabolomic method for XCMS data processing. Compared to the published parameters, the new XCMS parameters decrease the total number of features by 15% (a reduction in noise features) while increasing the number of true metabolites detected and grouped by 20%. Our credentialing platform relies on easily generated E. coli samples and a simple software algorithm that is freely available on our laboratory Web site (http://pattilab.wustl.edu/software/credential/). We have validated the credentialing platform with reversed-phase and hydrophilic interaction liquid chromatography as well as Agilent, Thermo Scientific, AB SCIEX, and LECO mass spectrometers. Thus, the credentialing platform can readily be applied by any laboratory to optimize their untargeted metabolomic pipeline for metabolite extraction, chromatographic separation, mass spectrometric detection, and bioinformatic processing.


Increased soluble CD14 serum levels and altered CD14 expression of peripheral blood monocytes in HIV-infected patients.
Serum levels of soluble CD14 were elevated in HIV-infected asymptomatic patients or those with lymphadenopathy (CDC II/III) 2.9 +/- 0.8 mg/l compared with normal controls with 2.2 +/- 0.47 mg/l, P < 0.001. A further rise was seen in patients with ARC (CDC IVA) 3.8 +/- 1.1 mg/l, P < 0.01 and patients with AIDS (CDC IVB-D) 5.7 +/- 2.5 mg/l, P < 0.01. Although absolute numbers of CD14+ cells decrease in the AIDS group, the percentage of CD14+ monocytes did not change. In contrast, levels of soluble T cell antigens sCD4 and sCD8, which are higher in HIV-infected patients compared with normal subjects, showed no increase with disease progression. Serum levels of sCD14 were correlated positively with beta 2-microglobulin levels (rs = 0.63, P < 0.0001). Whereas the percentage of CD14+ monocytes did not change, an increase in monocytic CD14 expression in HIV-infected patients was observed (P < 0.01). The percentage of a monocyte subset expressing both CD14 and CD16 increased from 6% in normal healthy persons to 13% in HIV-infected patients (P < 0.001), and did not vary between the HIV patient groups. Incubation of cultured peripheral blood monocytes with azidothymidine had no effect on either normal or LPS-induced or IL-4-inhibited sCD14 release in vitro. Therefore, an effect of AZT on sCD14 serum values in vivo is considered to be unlikely. Our data further provide evidence that monocytes/macrophages are engaged in HIV infection.


Using a teaching OSCE to prompt learners to engage with patients who talk about religion and/or spirituality.
The objective structured clinical examination (OSCE) has only occasionally been used as a teaching tool. The authors describe the initial use of an educational innovation consisting of a teaching OSCE used as "sensitizing practice," followed by personal, guided, and group reflection. Staff and resident physicians and one medical student (N = 28) at a community hospital's family medicine residency participated in the innovation during August 2012. The initial use of the educational innovation allowed learners to engage in a potentially challenging conversation with a standardized patient about religion and/or spirituality (R/S). The aim of the innovation was not to equip learners with a particular tactic to introduce or discuss R/S but, rather, to prompt learners to engage in mindful practice with patients who identify R/S as part of their biopsychosocial contexts. Written, dyadic, and group reflection added value to the OSCE by allowing participants to reflect on a difficult learning objective over time. Participants moved along the stages-of-change continuum when engaging in guided reflection compared with personal reflection. Additionally, all participants provided evidence of at least the preparation stage at the time of guided reflection. By following the OSCE's sensitizing practice with three periods of reflection, learners were enabled first, to recognize the need for readiness to address challenging communication topics (in this case, R/S) and, second, to reflect on practiced strategies for those conversations. The educational innovation can help learners become more aware of and skillful in dealing with difficult physician-patient communication topics.


Measuring the effect of inter-study variability on estimating prediction error.
The biomarker discovery field is replete with molecular signatures that have not translated into the clinic despite ostensibly promising performance in predicting disease phenotypes. One widely cited reason is lack of classification consistency, largely due to failure to maintain performance from study to study. This failure is widely attributed to variability in data collected for the same phenotype among disparate studies, due to technical factors unrelated to phenotypes (e.g., laboratory settings resulting in "batch-effects") and non-phenotype-associated biological variation in the underlying populations. These sources of variability persist in new data collection technologies. Here we quantify the impact of these combined "study-effects" on a disease signature's predictive performance by comparing two types of validation methods: ordinary randomized cross-validation (RCV), which extracts random subsets of samples for testing, and inter-study validation (ISV), which excludes an entire study for testing. Whereas RCV hardwires an assumption of training and testing on identically distributed data, this key property is lost in ISV, yielding systematic decreases in performance estimates relative to RCV. Measuring the RCV-ISV difference as a function of number of studies quantifies influence of study-effects on performance. As a case study, we gathered publicly available gene expression data from 1,470 microarray samples of 6 lung phenotypes from 26 independent experimental studies and 769 RNA-seq samples of 2 lung phenotypes from 4 independent studies. We find that the RCV-ISV performance discrepancy is greater in phenotypes with few studies, and that the ISV performance converges toward RCV performance as data from additional studies are incorporated into classification. We show that by examining how fast ISV performance approaches RCV as the number of studies is increased, one can estimate when "sufficient" diversity has been achieved for learning a molecular signature likely to translate without significant loss of accuracy to new clinical settings.


After-effects of repetitive anodal transcranial direct current stimulation on learning and memory in a rat model of Alzheimer's disease.
Repetitive anodal transcranial direct current stimulation (tDCS) in a rat model of Alzheimer's disease (AD) has been shown to have distinct neuroprotective effects. Moreover, the effects of anodal tDCS not only occur during the stimulation but also persist after the stimulation has ended (after-effects). Here, the duration of the after-effects induced by repetitive anodal tDCS was investigated based on our previous studies. Adult male Sprague-Dawley rats were divided into three groups: a sham group, a β-amyloid (Aβ) group (AD group) and a stimulation group (ATD group). Aβ was injected into the bilateral hippocampi of the rats in the AD and ATD groups to produce the AD model. Rats in the ATD group underwent 10 sessions of anodal tDCS, and the after-effects of repetitive anodal tDCS were evaluated by behavioral and histological analyses. A Morris water maze (MWM) was utilized on a monthly basis to assess spatial learning and memory abilities. The ATD group showed shorter escape latencies and more platform region crossings than the AD group. Hippocampal choline acetyltransferase (ChAT) and glial fibrillary acidic protein (GFAP) immunohistochemical analyses were carried out after the last MWM assessment. The immunohistochemistry results showed notable differences among the groups, particularly between the AD and ATD groups. This study reveals that repetitive anodal tDCS can not only improve cognitive function and memory performance but also has long-term after-effects that persist for 2 months.


MHC-disassortative mate choice and inbreeding avoidance in a solitary primate.
Sexual selection theory suggests that choice for partners carrying dissimilar genes at the major histocompatibility complex (MHC) may play a role in maintaining genetic variation in animal populations by limiting inbreeding or improving the immunity of future offspring. However, it is often difficult to establish whether the observed MHC dissimilarity among mates drives mate choice or represents a by-product of inbreeding avoidance based on MHC-independent cues. Here, we used 454-sequencing and a 10-year study of wild grey mouse lemurs (Microcebus murinus), small, solitary primates from western Madagascar, to compare the relative importance on the mate choice of two MHC class II genes, DRB and DQB, that are equally variable but display contrasting patterns of selection at the molecular level, with DRB under stronger diversifying selection. We further assessed the effect of the genetic relatedness and of the spatial distance among candidate mates on the detection of MHC-dependent mate choice. Our results reveal inbreeding avoidance, along with disassortative mate choice at DRB, but not at DQB. DRB-disassortative mate choice remains detectable after excluding all related dyads (characterized by a relatedness coefficient r > 0), but varies slightly with the spatial distance among candidate mates. These findings suggest that the observed deviations from random mate choice at MHC are driven by functionally important MHC genes (like DRB) rather than passively resulting from inbreeding avoidance and further emphasize the need for taking into account the spatial and genetic structure of the population in correlative tests of MHC-dependent mate choice.


Speciation and quantitative mapping of metal species in microbial biofilms using scanning transmission X-ray microscopy.
A scanning transmission X-ray microscope illuminated with synchrotron light was used to investigate the speciation and spatial distributions of metals in a microbial biofilm cultivated from river water. Metal 2p absorption edge signals were used to provide metal speciation (through shapes of the absorption spectra) and quantitative spatial distributions of the metal species. This paper presents sample data and describes methods for extracting quantitative maps of metal species from image sequences recorded in the region of the metal 2p edges. Comparisons were made with biochemical characterization of the same region using images recorded at the C 1s and O 1s edges. The method is applied to detailed quantitative analysis of ferrous and ferric iron in a river biofilm, in concert with mapping Ni(II) and Mn(II) species in the same region. The distributions of the metal species are discussed in the context of the biofilm structure. These results demonstrate that soft X-ray STXM measurements at the metal 2p absorption edges can be used to speciate metals and to provide quantitative spatial distribution maps for metal species in environmental samples with 50 nm spatial resolution.


Holographic interferometry of oil films and droplets in water with a single-beam mirror-type scheme.
Application of single-beam reflective laser optical interferometry for oil films and droplets in water detection and characterization is discussed. Oil films can be detected by the appearance of characteristic interference patterns. Analytical expressions describing intensity distribution in these interference patterns allow determination of oil film thickness, size of oil droplets, and distance to the oil film from the observation plane. Results from these analyses indicate that oil spill aging and breakup can be monitored in real time by analyzing time-dependent holographic fringe patterns. Interferometric methods of oil spill detection and characterization can be automated using digital holography with three-dimensional reconstruction of the time-changing oil spill topography. In this effort, the interferometric methods were applied to samples from Chevron oil and British Petroleum MC252 oil obtained during the Deep Water Horizon oil spill in the Gulf of Mexico.


Association of Epicardial Fat Tissue with Coronary Artery Disease and Left Ventricle Diastolic Function Indicators.
BACKGROUND Epicardial fat tissue (EAT) acts as brown adipose tissue and protects the heart and coronary arteries against hypothermia. Recent studies demonstrated that EAT is a source of both anti-inflammatory and atherogenic cytokines. In this study, our aim was to investigate the association of vertical, horizontal, and area measurements of EAT thickness and their association with coronary artery disease, diastolic function, and myocardial performance index in patients who underwent coronary angiography. MATERIAL AND METHODS The study population consisted of patients who presented to our outpatient clinic with chest pain and whose non-invasive stress tests were positive between June 2015 and July 2017. Echocardiographic examinations were performed prior to the angiography. Coronary angiograms were performed using Judkins method from the femoral artery. RESULTS Mean vertical thickness of EAT was 0.6 cm in patients with CAD and 0.46 cm in those without CAD (p=0.0001). Mean horizontal length of EAT was 2.91 cm in patients with CAD and was 2.41 cm in the subjects without CAD (p=0.001). ROC analysis showed 81% sensitivity and 53% specificity for a cut-off value of 0.45, and 67% sensitivity and 71% specificity for a cut-off value of 0.55 for EAT vertical (cm). Multivariate analysis showed that EAT is an independent risk factor for coronary artery disease. CONCLUSIONS Echocardiography is an inexpensive routine assessment for most patients. EAT thickness determined by echocardiography may be a useful indicator of increased CAD risk, but not diastolic dysfunction, of the left ventricle.


Bovine viral diarrhea virus NS3 serine proteinase: polyprotein cleavage sites, cofactor requirements, and molecular model of an enzyme essential for pestivirus replication.
Members of the Flaviviridae encode a serine proteinase termed NS3 that is responsible for processing at several sites in the viral polyproteins. In this report, we show that the NS3 proteinase of the pestivirus bovine viral diarrhea virus (BVDV) (NADL strain) is required for processing at nonstructural (NS) protein sites 3/4A, 4A/4B, 4B/5A, and 5A/5B but not for cleavage at the junction between NS2 and NS3. Cleavage sites of the proteinase were determined by amino-terminal sequence analysis of the NS4A, NS4B, NS5A, and NS5B proteins. A conserved leucine residue is found at the P1 position of all four cleavage sites, followed by either serine (3/4A, 4B/5A, and 5A/5B sites) or alanine (4A/4B site) at the P1' position. Consistent with this cleavage site preference, a structural model of the pestivirus NS3 proteinase predicts a highly hydrophobic P1 specificity pocket. trans-Processing experiments implicate the 64-residue NS4A protein as an NS3 proteinase cofactor required for cleavage at the 4B/5A and 5A/5B sites. Finally, using a full-length functional BVDV cDNA clone, we demonstrate that a catalytically active NS3 serine proteinase is essential for pestivirus replication.


Explanatory models of stroke in Ghana: perspectives of stroke survivors and their caregivers.
Objective: This study examines explanatory models (EMs) of stroke and its complications among people living with stroke, and their caregivers, in two urban poor communities in Accra (Ga Mashie) and Korle Bu Teaching Hospital (KBTH), Accra.Methods: Twenty-two stroke survivors and 29 caregivers were recruited from 2 urban poor communities in Accra and KBTH. Qualitative data were obtained using semi-structured interviews that lasted between 45 minutes and 2 hours. The interviews were audiotaped, transcribed and analysed thematically, informed by the concept of EMs of illness.Results: Participants referred to stroke as a sudden event and they expressed different emotional responses after the stroke onset. Stroke survivors and their caregivers attributed stroke with poor lifestyle practices, high blood pressure, unhealthy diet and dietary practices, supernatural causes, stress, family history, other chronic diseases, and delay in treatment of symptoms. While the stroke survivors associated stroke complications with physical disability and stigmatisation, the caregivers associated these with physical disability, behavioural and psychological changes, cognitive disability and death. These associations were mostly influenced by the biomedical model of stroke.Conclusion: The biomedical model of stroke is important for developing interventions that will be accepted by the stroke survivors and the caregivers. Nevertheless, sociocultural explanations of stroke need to be taken into consideration during delivery of medical information to the participants. This study proposes an integrated biopsychosociocultural approach for stroke intervention among the study participants.


Parameterization of connectionist models.
We present a method for estimating parameters of connectionist models that allows the model's output to fit as closely as possible to empirical data. The method minimizes a cost function that measures the difference between statistics computed from the model's output and statistics computed from the subjects' performance. An optimization algorithm finds the values of the parameters that minimize the value of this cost function. The cost function also indicates whether the model's statistics are significantly different from the data's. In some cases, the method can find the optimal parameters automatically. In others, the method may facilitate the manual search for optimal parameters. The method has been implemented in Matlab, is fully documented, and is available for free download from the Psychonomic Society Web archive at www.psychonomic.org/archive/.


Biomarkers increase detection of active smoking and secondhand smoke exposure in critically ill patients.
The association between tobacco smoke exposure and critical illness is not well studied, largely because obtaining an accurate smoking history from critically ill patients is difficult. Biomarkers can provide quantitative data on active and secondhand cigarette smoke exposure. We sought to compare cigarette smoke exposure as measured by biomarkers to exposure by self-report in a cohort of critically ill patients and to determine how well biomarkers of cigarette smoke exposure correlate with each other in this population. Serum and urine cotinine and trans-3'-hydroxycotinine, urine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol, and hair and nail nicotine levels were measured in 60 subjects enrolled in an observational cohort of critically ill subjects at a tertiary academic medical center in Tennessee. Smoking history was obtained from patients, their surrogates, or the medical chart. Cigarette smoke exposure as measured by biomarkers was compared to exposure by history. By smoking history, 29 subjects were identified as smokers, 28 were identified as nonsmokers, and 3 were identified as unknown. The combination of serum cotinine and urine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol identified 27 of the 28 nonsmokers by history either as active smokers (n = 6, 21%) or as exposed to secondhand smoke (n = 21, 75%). All biomarker levels were strongly correlated with each other (r = .69-.95, p < .0001). The combination of serum cotinine and urine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol identified considerably more active smokers than did smoking history and detected a high prevalence of secondhand smoke exposure in a critically ill population. These markers will be important for future studies investigating the relationship between active smoking and secondhand smoke exposure and critical illness.


Teaching Happiness to Teachers - Development and Evaluation of a Training in Subjective Well-Being.
Teachers' health is a persistent challenge for educational systems all over the world. Moreover, research results - especially in the domain of positive psychology - indicate that high levels of well-being are associated with additional benefits improving teachers' professional performance. Therefore, a training to foster subjective well-being with one training day, two booster sessions, and exercises before, during, and after the meetings was developed. It consisted of about 10 h of face-to-face time and about 3 h for the exercises in total over a 5-week training period. Main contents were conditions and consequences of positive and negative emotions and well-being, emotion regulation, time management, savoring and gratitude and the application of positive psychological interventions (like Three Good Things). Analyses of planned contrasts by means of a waiting control group design with three measurement points (pre, post, and follow-up) showed a significantly higher increase for the training group (n = 42) than for the control group (n = 47) in the frequency of positive emotions, life satisfaction, and flourishing (interaction effects d = 0.44, d = 0.31, and d = 0.32) and a significantly stronger decrease in the frequency of negative emotions, perceived stress, and experiencing emotional exhaustion (interaction effects d = 0.69, d = 0.51, and d = 0.47) from pre to 1-month follow-up. Training effects were also visible up to 5 months, although no control group could be realized for this period due to the field approach.


An operando emission spectroscopy study of Pt/Al2O3 and Pt/CeO2/Al2O3.
In situ time-resolved spectroscopic examination of catalysts based on well dispersed nanoparticles on metal oxides under transient conditions significantly facilitates the elucidation of reaction mechanisms. In this contribution, we demonstrate the level of structural information that can be obtained using high-energy resolution off-resonant spectroscopy (HEROS) to study 1.3 wt% Pt/Al2O3 and 1.3 wt% Pt/20 wt% CeO2/Al2O3 catalysts subjected to redox pulsing. First, HEROS is compared with XANES in a temperature programmed reduction experiment to demonstrate the increased sensitivity and time resolution of HEROS. Second, modulation excitation spectroscopy is exploited by redox pulsing to enhance the sensitivity of HEROS to structural changes by the application of phase sensitive detection (PSD) to the time-resolved HEROS data set. The HEROS measurements were complemented by resonant X-ray emission (RXES) and diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy measurements performed under identical conditions and in a single reactor cell in order to probe different aspects of the catalyst materials under the selected experimental conditions.


Metabolic, hormonal, and vascular changes after synthetic oestrogen therapy in oophorectomized women.
Mestranol, the three-methyl ether form of ethinyloestradiol and one of the two oestrogens used in oral contraceptive steroids, was administered in a dose of 0.02 mg daily for 120 days to 25 oophorectomized women.Urinary oestriol and pregnanediol excretions were unaffected by the mestranol treatment but there was a shift of the maturation index of the vaginal smear to the right, indicating a correction of the pretreatment oestrogen deficiency. No significant change in the blood pressure or electrocardiograph recordings occurred during this relatively short period of administration. A significant rise in the serum protein-bound iodine, which might be regarded as an undesirable effect of mestranol on a long-term basis, occurred. Hepatic function as measured by bromsulphthalein was not impaired by the treatment. Mestranol had no effect on the total body water or on the total exchangeable potassium of the women. Its two most serious adverse effects were impairment of glucose tolerance and a high incidence (16%) of venous thrombo-embolic disease.The gravity of the adverse effects far outweighs any beneficial ones and precludes the use of mestranol alone for long-term hormone replacement therapy in postmenopausal women.


Probiotics as a complementary therapeutic approach in nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD) is currently recognized as one of the most common causes of chronic liver disease. It involves a spectrum of conditions that include pure steatosis without inflammation, steatohepatitis, fibrosis and cirrhosis. The key factor in the pathophysiology of NAFLD is insulin resistance that determines lipid accumulation in the hepatocytes and, thus, oxidative stress, which is followed by inflammatory response. However, NAFLD pathogenesis is still largely unknown and has been extensively investigated. Although life style modification with the aim of losing weight has been advocated to treat this disorder, its effectiveness is limited; additionally, there is no specific pharmacologic treatment until nowadays. Recent evidence suggests that the gut microbiota may play a role in the development of insulin resistance, hepatic steatosis, necroinflammation and fibrosis. Differences in gut microbiota between NAFLD patients and lean individuals as well as presence of small intestinal bacterial overgrowth in NAFLD subjects have been demonstrated. Furthermore, some data indicate that the immunoregulatory effects of probiotics may be beneficial in NAFLD treatment as they modulate the intestinal microbiota; improve epithelial barrier function and strengthen the intestinal wall decreasing its permeability; reduce bacterial translocation and endotoxemia; improve intestinal inflammation; and reduce oxidative and inflammatory liver damage. In this article, we review the clinical trials on the use of probiotics in the treatment of NAFLD and discuss the effects of these agents and their efficacy as an emerging therapeutic resource to treat NAFLD patients.


Tricuspid valve endocarditis.
Right-sided infective endocarditis (RSIE) is less common than left-sided infective endocarditis (IE), encompassing only 5-10% of cases of IE. Ninety percent of RSIE involves the tricuspid valve (TV). Given the relatively small numbers of TVIE cases operated on at most institutions, the purpose of this review is to highlight and discuss the current understanding of IE involving the TV. RSIE and TVIE are strongly associated with intravenous drug use (IVDU), although pacemaker leads, defibrillator leads and vascular access for dialysis are also major risk factors. Staphylococcus aureus is the predominant causative organism in TVIE. Most patients with TVIE are successfully treated with antibiotics, however, 5-16% of RSIE cases eventually require surgical intervention. Indications and timing for surgery are less clear than for left-sided IE; surgery is primarily considered for failed medical therapy, large vegetations and septic pulmonary embolism, and less often for TV regurgitation and heart failure. Most patients with an infected prosthetic TV will require surgery. Concomitant left-sided IE has its own surgical indications. Earlier surgical intervention may potentially prevent further destruction of leaflet tissue and increase the likelihood of TV repair. Fortunately, TV debridement and repair can be accomplished in most cases, even those with extensive valve destruction, using a variety of techniques. Valve repair is advocated over replacement, particularly in IVDUs patients who are young, non-compliant and have a higher risk of recurrent infection and reoperation with valve replacement. Excising the valve without replacing, it is not advocated; it has been reported previously, but these patients are likely to be symptomatic, particularly in cases with septic pulmonary embolism and increased pulmonary vascular resistance. Patients with concomitant left-sided involvement have worse prognosis than those with RSIE alone, due predominantly to greater likelihood of invasion and abscess formation in left-sided IE. Patients with isolated TVIE have an operative mortality between 0-15% and excellent survival.


Extracellular glutamate alters mature osteoclast and osteoblast functions.
Glutamatergic intercellular communication is involved in many aspects of metabolic homeostasis in normal bone. In bone metastasis, the balance between bone formation and degradation is disrupted. Although the responsible mechanisms are not clear, we have previously identified that cancer cell lines used in bone tumour models secrete glutamate, suggesting that tumour-derived glutamate may disrupt sensitive signalling systems in bone. This study examines the role of glutamate in mature osteoclastic bone resorption, osteoblast differentiation, and bone nodule formation. Glutamate was found to have no effect on the survival or activity of mature osteoclasts, although glutamate transporter inhibition and receptor blockade increased the number of bone resorption pits. Furthermore, transporter inhibition increased the area of resorbed bone while significantly decreasing the number of osteoclasts. Alkaline phosphatase activity and extracellular matrix mineralization were used as measurements of osteoblast differentiation. Glutamate significantly increased osteoblast differentiation and mineralization, but transport inhibitors had no effect. These studies support earlier findings suggesting that glutamate may be more important for osteoclastogenesis than for osteoclast proliferation or functions. Since glutamate is capable of changing the differentiation and activities of both osteoclast and osteoblast cell types in bone, it is reasonable to postulate that tumour-derived glutamate may impact bone homeostasis in bone metastasis.


Nonlinearly limited saturable-absorber mode locking of an erbium fiber laser.
We describe an erbium fiber laser that is passively mode locked by a novel, precision antireflection-coated semiconductor saturable-absorber mirror that incorporates an additional two-photon absorber. It is shown that passive mode locking evolves from a Q-switching instability. The results are achieved by use of saturable absorbers that provide a large (15%) nonlinear (saturable) loss. Exploiting two-photon absorption can substantially reduce the peak power of the Q-switched pulses, which results in improved reliability of the laser. Moreover, two-photon absorption can be used to produce an optimal stability range for saturable-absorber mode locking.


Caught in the middle: the contested politics of HIV/AIDS and health policy in Vietnam.
Drawing on the changing landscape of responses to HIV in Vietnam, this article describes the key players and analyzes the relationships between global players and local interests, including both the omnipresent state and an emerging civil society presence. We discuss the critical importance of timing for policy intervention and the role of health policy in shaping the broader social terrain. The interventions of external actors such as the US President's Emergency Plan for AIDS Relief (PEPFAR) and the Global Fund were instrumental in improving both policies and programs at a critical juncture, when the national responses to the epidemic had been ineffective. At the same time, those global interventions met resistance and led to unintended consequences, both welcome and unwelcome. Furthermore, the looming specter of donor withdrawal and the very gradually emerging national ownership raise many questions about capacity for scale-up and sustainability of the significant achievements to date. Further monitoring and in-depth analysis of the Vietnamese responses to the HIV epidemic in the next few years or so, we contend, have the potential to provide unique insights into the challenges faced by developing countries caught in the complex webs of health politics and policies at both the global and the national levels.


Effect of water and saliva contamination on shear bond strength of brackets bonded with conventional, hydrophilic, and self-etching primers.
This study assessed the effect of water and saliva contamination on the shear bond strength and bond failure site of 3 different orthodontic primers (Transbond XT, Transbond Moisture Insensitive Primer, and Transbond Plus Self Etching Primer; 3M Unitek, Monrovia, Calif) used with a light-cured composite resin (Transbond XT). Bovine permanent mandibular incisors (315) were randomly divided into 21 groups (15 in each group). Each primer-adhesive combination was tested under 7 different enamel surface conditions: (1) dry, (2) water application before priming, (3) water application after priming, (4) water application before and after priming, (5) saliva application before priming, (6) saliva application after priming, and (7) saliva application before and after priming. Stainless steel brackets were bonded in each test group with composite resin. After bonding, all samples were stored in distilled water at room temperature for 24 hours and then tested for shear bond strength. Noncontaminated enamel surfaces had the highest bond strengths for conventional, hydrophilic, and self-etching primers, which produced the same strength values. In most contaminated conditions, the self-etching primer had higher strength values than either the hydrophilic or conventional primers. The self-etching primer was the least influenced by water and saliva contamination, except when moistening occurred after the recommended 3-second air burst. No significant differences in debond locations were found among the groups bonded with the self-etching primer under the various enamel conditions.


Formation of a new type of uranium(iv) poly-oxo cluster {U38} based on a controlled release of water via esterification reaction.
A new strategy for the synthesis of large poly-oxo clusters bearing 38 tetravalent uranium atoms {U38} has been developed by controlling the water release from the esterification reaction between a carboxylic acid and an alcohol. The molecular entity [U38O56Cl40(H2O)2(ipa)20]·(ipa) x (ipa = isopropanol) was crystallized from the solvothermal reaction of a mixture of UCl4 and benzoic acid in isopropanol at temperature ranging from 70 to 130 °C. Its crystal structure reveals the molecular assembly of the UO2 fluorite-like inner core {U14} with oxo groups bridging the uranium centers. The {U14} core is further surrounded by six tetrameric sub-units of {U4} to form the {U38} cluster. Its surface is decorated by either bridging- and terminal chloride anions or terminal isopropanol molecules. Another synthesis using the same reactant mixture at room temperature resulted in the crystallization of a discrete dinuclear complex [U2Cl4(bz)4(ipa)4]·(ipa)0.5 (bz = benzoate), in which each uranium center is coordinated by two chlorine atoms, four oxygen atoms from carboxylate groups and two additional oxygen atoms from isopropanol. The slow production of water released from the esterification of isopropanol allows the formation of the giant cluster with oxo bridges linking the uranium atoms at a temperature above 70 °C, whereas no such oxo groups are present in the dinuclear complex formed at room temperature. The kinetics of {U38} crystallization as well as the ester formation are analyzed and discussed. SAXS experiments indicate that the {U38} species are not dominant in the supernatant, but hexanuclear entities which are closely related to the [U6O8] type are formed.


Maturation of Complex Synaptic Connections of Layer 5 Cortical Axons in the Posterior Thalamic Nucleus Requires SNAP25.
Synapses are able to form in the absence of neuronal activity, but how is their subsequent maturation affected in the absence of regulated vesicular release? We explored this question using 3D electron microscopy and immunoelectron microscopy analyses in the large, complex synapses formed between cortical sensory efferent axons and dendrites in the posterior thalamic nucleus. Using a Synaptosome-associated protein 25 conditional knockout (Snap25 cKO), we found that during the first 2 postnatal weeks the axonal boutons emerge and increase in the size similar to the control animals. However, by P18, when an adult-like architecture should normally be established, axons were significantly smaller with 3D reconstructions, showing that each Snap25 cKO bouton only forms a single synapse with the connecting dendritic shaft. No excrescences from the dendrites were formed, and none of the normally large glomerular axon endings were seen. These results show that activity mediated through regulated vesicular release from the presynaptic terminal is not necessary for the formation of synapses, but it is required for the maturation of the specialized synaptic structures between layer 5 corticothalamic projections in the posterior thalamic nucleus.


Enrichment and Proteomic Characterization of the Cyst Wall from In Vitro Toxoplasma gondii Cysts.
The tissue cyst of Toxoplasma gondii, found in latent infection, serves a critical role in both transmission and reactivation of this organism. Within infected cells, slowly replicating parasites (bradyzoites) are surrounded by a cyst matrix, cyst wall, and cyst membrane. The cyst wall is clearly delineated by ultrastructural analysis; however, the composition and function of this layer in host-parasite interactions are not fully understood. In order to understand the composition of the cyst wall, a proteomic analysis of purified cyst wall fragments, that were enriched with Percoll gradients and subsequently immunoprecipitated with CST1 antibody, was performed. Known cyst wall proteins, such as CST1, BPK1, MCP4, MAG1, GRA2, GRA3, and GRA5, were identified in this preparation by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, dense granule proteins (GRAs) not previously shown to associate with the cyst wall, as well as uncharacterized hypothetical proteins, were identified in this cyst wall preparation. Several of these hypothetical cyst wall (CST) proteins were epitope tagged, and immunofluorescence assays confirmed their localization as novel cyst matrix and cyst wall proteins. Expression of two of these newly identified cyst wall proteins was eliminated by gene knockout (CST2-KO and CST3-KO). CST2-KO parasites were highly attenuated in virulence and did not establish detectable cyst burdens. This targeted proteomic approach allowed the identification of new components of the cyst wall that probably have roles in the parasite/host interface.IMPORTANCEToxoplasma gondii is a highly prevalent parasite worldwide that presents life-threatening risks to immunocompromised and pregnant individuals. Whereas the life stage responsible for acute infection can be treated, the life stage responsible for chronic infection is refractory to currently available therapeutics. Little is known about the protein composition of the cyst wall, an amorphous structure formed by parasites that is suspected to facilitate persistence within muscle and nervous tissue during chronic (latent) infection. By implementing a refined approach to selectively purify cyst wall fragments, we identified several known and novel cyst wall proteins from our sample preparations. We confirmed the localizations of several proteins from this data set and identified one that is involved in parasite virulence. These data will propel further studies on cyst wall structure and function, leading to therapeutic strategies that can eliminate the chronic infection stage.


1,25(OH)2D3 alters growth plate maturation and bone architecture in young rats with normal renal function.
Whereas detrimental effects of vitamin D deficiency are known over century, the effects of vitamin D receptor activation by 1,25(OH)(2)D(3), the principal hormonal form of vitamin D, on the growing bone and its growth plate are less clear. Currently, 1,25(OH)(2)D(3) is used in pediatric patients with chronic kidney disease and mineral and bone disorder (CKD-MBD) and is strongly associated with growth retardation. Here, we investigate the effect of 1,25(OH)(2)D(3) treatment on bone development in normal young rats, unrelated to renal insufficiency. Young rats received daily i.p. injections of 1 µg/kg 1,25(OH)(2)D(3) for one week, or intermittent 3 µg/kg 1,25(OH)(2)D(3) for one month. Histological analysis revealed narrower tibial growth plates, predominantly in the hypertrophic zone of 1,25(OH)(2)D(3)-treated animals in both experimental protocols. This phenotype was supported by narrower distribution of aggrecan, collagens II and X mRNA, shown by in situ hybridization. Concomitant with altered chondrocyte maturation, 1,25(OH)(2)D(3) increased chondrocyte proliferation and apoptosis in terminal hypertrophic cells. In vitro treatment of the chondrocytic cell line ATDC5 with 1,25(OH)(2)D(3) lowered differentiation and increased proliferation dose and time-dependently. Micro-CT analysis of femurs from 1-week 1,25(OH)(2)D(3)-treated group revealed reduced cortical thickness, elevated cortical porosity, and higher trabecular number and thickness. 1-month administration resulted in a similar cortical phenotype but without effect on trabecular bone. Evaluation of fluorochrome binding with confocal microscopy revealed inhibiting effects of 1,25(OH)(2)D(3) on intracortical bone formation. This study shows negative effects of 1,25(OH)(2)D(3) on growth plate and bone which may contribute to the exacerbation of MBD in the CKD pediatric patients.


Adaptative biochemical pathways and regulatory networks in Klebsiella oxytoca BAS-10 producing a biotechnologically relevant exopolysaccharide during Fe(III)-citrate fermentation.
A bacterial strain previously isolated from pyrite mine drainage and named BAS-10 was tentatively identified as Klebsiella oxytoca. Unlikely other enterobacteria, BAS-10 is able to grow on Fe(III)-citrate as sole carbon and energy source, yielding acetic acid and CO2 coupled with Fe(III) reduction to Fe(II) and showing unusual physiological characteristics. In fact, under this growth condition, BAS-10 produces an exopolysaccharide (EPS) having a high rhamnose content and metal-binding properties, whose biotechnological applications were proven as very relevant. Further phylogenetic analysis, based on 16S rDNA sequence, definitively confirmed that BAS-10 belongs to K. oxytoca species. In order to rationalize the biochemical peculiarities of this unusual enterobacteriun, combined 2D-Differential Gel Electrophoresis (2D-DIGE) analysis and mass spectrometry procedures were used to investigate its proteomic changes: i) under aerobic or anaerobic cultivation with Fe(III)-citrate as sole carbon source; ii) under anaerobic cultivations using Na(I)-citrate or Fe(III)-citrate as sole carbon source. Combining data from these differential studies peculiar levels of outer membrane proteins, key regulatory factors of carbon and nitrogen metabolism and enzymes involved in TCA cycle and sugar biosynthesis or required for citrate fermentation and stress response during anaerobic growth on Fe(III)-citrate were revealed. The protein differential regulation seems to ensure efficient cell growth coupled with EPS production by adapting metabolic and biochemical processes in order to face iron toxicity and to optimize energy production. Differential proteomics provided insights on the molecular mechanisms necessary for anaeorobic utilization of Fe(III)-citrate in a biotechnologically promising enterobacteriun, also revealing genes that can be targeted for the rational design of high-yielding EPS producer strains.


In utero diethylstilbestrol (DES) exposure alters Hox gene expression in the developing müllerian system.
Diethylstilbestrol (DES) was widely used to treat pregnant women through 1971. The reproductive tracts of their female offspring exposed to DES in utero are characterized by anatomic abnormalities. Here we show that DES administered to mice in utero produces changes in the expression pattern of several Hox genes that are involved in patterning of the reproductive tract. DES produces posterior shifts in Hox gene expression and homeotic anterior transformations of the reproductive tract. In human uterine or cervical cell cultures, DES induces HOXA9 or HOXA10 gene expression, respectively, to levels approximately twofold that induced by estradiol. The DES-induced expression is not inhibited by cyclohexamide. Estrogens are novel morphogens that directly regulate the expression pattern of posterior Hox genes in a manner analogous to retinoic acid regulation of anterior Hox genes. Alterations in HOX gene expression are a molecular mechanism by which DES affects reproductive tract development. Changes in Hox gene expression are a potential marker for the effects of in utero drug use that may become apparent only at late stages of development.


Organizing an efficient trauma team.
A prospective analysis of 207 trauma patients, from three internationally recognized trauma centres, showed that trauma teams in which staff carry out allocated tasks simultaneously have the quickest resuscitation times. This finding was further tested by introducing these changes into a fourth centre. A comparison of resuscitation stage times was made in 26 patients before and 24 patients after the introduction of the organizational changes. Significant time reductions were found in all the stages, except the time taken to examine the patient. The time taken to complete the resuscitation was reduced by over half from 122 to 56 min. Significant time reductions applied even when variations in the type of patient, the team size or seniority of the team leader was taken into account. Recommendations for the organization of trauma teams are made.


[Vascularization of the skull base--review of the literature and illustrative cases].
The dural vascularization of the skull base and proximal nervous structures, depends on the external carotid artery, internal carotid artery and vertebral artery branches. The authors describe these branches, emphasizing possible anastomoses between them, and present images from their own experience. Such arterio-arterial anastomosis represent incomplete regressions of embryologic arterial connections and are always present, but functionally inactive. It is usually in the presence of pathologic situations (stroke, tumours, dural fistulas) that they become active and, therefore, special care is necessary if endovascular procedures with embolic material are to be performed.


Preference effects on friendship choice: Evidence from an online field experiment.
Observed friendship choices are constrained by social structures and thus problematic indicators for underlying personal preferences. In this paper, we report on a study demonstrating the causal effects of preference in friendship choice based on an online field experiment. Specifically, we tested two important forces that govern friendship choices: preference for shared group identity (operationalized as the desire to befriend others sharing the same place-of-origin identity) and preference for high status (operationalized as the desire to befriend others from high-status institutions). Using an online field experiment in one of the largest social network service websites in China, we investigated the causal preference effects of these two forces free from structural constraints. The results of our study confirm the preference effects on friendship choice in both of the two dimensions we tested.


Prevalence of human metapneumovirus in adults with acute respiratory tract infection in Beijing, China.
To evaluate the prevalence and clinical manifestations of human metapneumovirus (hMPV) in immunocompetent Chinese adults with acute respiratory tract infections (ARTIs). A reverse transcription PCR (RT-PCR) assay targeting the P gene was developed in this study and used to detect hMPV in nasal and throat swabs collected from 2936 immunocompetent adult patients with ARTIs in Beijing, China between July 2008 and June 2010. Among the 2936 patients studied, 49 (1.7%) were positive for hMPV, of whom 14 (28.6%) were positive for hMPV_A2b, 19 (38.8%) for hMPV_B1, and 16 (32.6%) for hMPV_B2. hMPV_A1 was not detected. An average detection rate of 6.6% was observed in the peak months of the two epidemic seasons studied. The hMPV prevalence was higher in the sampled elderly (>65 years, 3.2%) than in middle aged adults (25-65 years; 2.0%) and teenagers (14-25 years; 0.9%). During the study period, hMPV infections showed a biennial rhythm of seasonality, peaking from November to March in 2008/09 and from March to June in 2010. hMPV infection plays an important role in immunocompetent adults in its epidemic season. The demographic and clinical data presented in this study improves our understanding of the pathogenesis and clinical burden of hMPV infection in adults.


Community water fluoridation predicts increase in age-adjusted incidence and prevalence of diabetes in 22 states from 2005 and 2010.
Community water fluoridation is considered a significant public health achievement of the 20th century. In this paper, the hypothesis that added water fluoridation has contributed to diabetes incidence and prevalence in the United States was investigated. Panel data from publicly available sources were used with population-averaged models to test the associations of added and natural fluoride on the outcomes at the county level in 22 states for the years 2005 and 2010. The findings suggest that a 1 mg increase in the county mean added fluoride significantly positively predicts a 0.23 per 1,000 person increase in age-adjusted diabetes incidence (P < 0.001), and a 0.17% increase in age-adjusted diabetes prevalence percent (P < 0.001), while natural fluoride concentration is significantly protective. For counties using fluorosilicic acid as the chemical additive, both outcomes were lower: by 0.45 per 1,000 persons (P < 0.001) and 0.33% (P < 0.001), respectively. These findings are adjusted for county-level and time-varying changes in per capita tap water consumption, poverty, year, population density, age-adjusted obesity and physical inactivity, and mean number of years since water fluoridation started. Sensitivity analyses revealed robust effects for both types of fluoride. Community water fluoridation is associated with epidemiological outcomes for diabetes.


The aperiodic exponent of subthalamic field potentials reflects excitation/inhibition balance in Parkinsonism.
Periodic features of neural time-series data, such as local field potentials (LFPs), are often quantified using power spectra. While the aperiodic exponent of spectra is typically disregarded, it is nevertheless modulated in a physiologically relevant manner and was recently hypothesised to reflect excitation/inhibition (E/I) balance in neuronal populations. Here, we used a cross-species in vivo electrophysiological approach to test the E/I hypothesis in the context of experimental and idiopathic Parkinsonism. We demonstrate in dopamine-depleted rats that aperiodic exponents and power at 30-100 Hz in subthalamic nucleus (STN) LFPs reflect defined changes in basal ganglia network activity; higher aperiodic exponents tally with lower levels of STN neuron firing and a balance tipped towards inhibition. Using STN-LFPs recorded from awake Parkinson's patients, we show that higher exponents accompany dopaminergic medication and deep brain stimulation (DBS) of STN, consistent with untreated Parkinson's manifesting as reduced inhibition and hyperactivity of STN. These results suggest that the aperiodic exponent of STN-LFPs in Parkinsonism reflects E/I balance and might be a candidate biomarker for adaptive DBS.


Pathogenesis of cancrum oris (noma): confounding interactions of malnutrition with infection.
This study showed that impoverished Nigerian children at risk for cancrum oris (noma) had significantly reduced plasma concentrations of zinc (< 10.8 micromol/L), retinol (< 1.05 micromol/L), ascorbate (< 11 micromol/L), and the essential amino acids, with prominently increased plasma and saliva levels of free cortisol, compared with their healthy counterparts. The nutrient deficiencies, in concert with previously reported widespread viral infections (measles, herpesviruses) in the children, would impair oral mucosal immunity. We postulate, subject to additional studies, that evolution of the oral mucosal ulcers including acute necrotizing gingivitis to noma is triggered by a consortium of microorganisms of which Fusobacterium necrophorum is a key component. Fusobacterium necrophorum elaborates several dermonecrotic toxic metabolites and is acquired by the impoverished children via fecal contamination resulting from shared residential facilities with animals and very poor environmental sanitation.


Studying Smad2 intranuclear diffusion dynamics by mathematical modelling of FRAP experiments.
We combine Fluorescence Recovery After Photobleaching (FRAP) experiments with mathematical modelling to study the dynamics inside the nucleus of both the TGF-β-sensitive transcriptional regulator Smad2, and Green-Fluorescent Protein (GFP). We show how combining modelling with bleaching strips of different areas allows a rigorous test of whether or not a protein is moving via diffusion as a single species. As noted recently by others, it is important to consider diffusion during the bleaching process. Neglecting it can cause serious error. Also, it is possible to use the bleaching process itself to provide an extra consistency test to the models predicting the recovery. With our method we show that the dynamics of GFP are consistent with it diffusing as a single species in a uniform environment in which flow is negligible. In contrast, the dynamics of the intracellular signal transducer Smad2 are never consistent with it moving as a single species via simple diffusion in a homogeneous environment without flow. Adding TGF-β slows down the dynamics of Smad2 but even without TGF-β, the Smad2 dynamics are influenced by one or more of: association, flow, and inhomogeneity in space of the dynamics. We suggest that the dynamics inside cells of many proteins may be poorly described by simple diffusion of a single species, and that our methodology provides a general and powerful way to test this hypothesis.


An Estimate of Biogenic Emissions of Volatile Organic Compounds during Summertime in China (7 pp).
and Aim. An accurate estimation of biogenic emissions of VOC (volatile organic compounds) is necessary for better understanding a series of current environmental problems such as summertime smog and global climate change. However, very limited studies have been reported on such emissions in China. The aim of this paper is to present an estimate of biogenic VOC emissions during summertime in China, and discuss its uncertainties and potential areas for further investigations. This study was mainly based on field data and related research available so far in China and abroad, including distributions of land use and vegetations, biomass densities and emission potentials. VOC were grouped into isoprene, monoterpenes and other VOC (OVOC). Emission potentials of forests were determined for 22 genera or species, and then assigned to 33 forest ecosystems. The NCEP/NCAR reanalysis database was used as standard environmental conditions. A typical summertime of July 1999 was chosen for detailed calculations. The biogenic VOC emissions in China in July were estimated to be 2.3×1012gC, with 42% as isoprene, 19% as monoterpenes and 39% as OVOC. About 77.3% of the emissions are generated from forests and woodlands. The averaged emission intensity was 4.11 mgC m-2 hr-1 for forests and 1.12 mgC m-2 hr-1 for all types of vegetations in China during the summertime. The uncertainty in the results arose from both the data and the assumptions used in the extrapolations. Generally, uncertainty in the field measurements is relatively small. A large part of the uncertainty mainly comes from the taxonomic method to assign emission potentials to unmeasured species, while the ARGR method serves to estimate leaf biomass and the emission algorithms to describe light and temperature dependence. This study describes a picture of the biogenic VOC emissions during summertime in China. Due to the uneven spatial and temporal distributions, biogenic VOC emissions may play an important role in the tropospheric chemistry during summertime. . Further investigations are needed to reduce uncertainties involved in the related factors such as emission potentials, leaf biomass, species distribution as well as the mechanisms of the emission activities. Besides ground measurements, attention should also be placed on other techniques such as remote-sensing and dynamic modeling. These new approaches, combined with ground measurements as basic database for calibration and evaluation, can hopefully provide more comprehensive information in the research of this field.


Molecular analysis of eight mutations in FBN1.
Mutations in the gene encoding extracellular glycoprotein fibrillin-1 (FBN1) cause Marfan syndrome (MFS) and other related connective tissue disorders. In this study, eight mutations have been detected in MFS patients by heteroduplex analysis. These comprise two missense mutations, C1835Y and C2258Y in calcium-binding epidermal growth factor-like domains, two nonsense mutations, R1541X and R2394X in transforming growth factor beta1-binding protein-like domains, one splice site mutation, which has been detected previously, and three small insertions or deletions resulting in a frameshift. Fibroblast cells have been established from seven of the MFS patients and the biochemical effects of the mutations on fibrillin-1 synthesis and secretion assessed by pulse-chase analysis. Each cysteine mutation resulted in the delayed secretion of fibrillin-1 and both nonsense and frameshift mutations caused reduced levels of synthesis and/or deposition of fibrillin-1. Indirect immunofluorescence and rotary shadowing electron microscopy analysis of fibrillin microfibrils revealed no major differences between normal and patient samples. We discuss the relative merits of the biochemical techniques used in this study.


Dietary approaches to positively influence fetal determinants of adult health.
An imbalance between production and scavenging of oxidants is a commonality found in factors that result in fetal determinants that negatively affect adult health. We reasoned that a dietary intervention that promotes oxidant scavenging through phase 2 protein induction would have positive effects on fetal programming of adult health. Previously, we demonstrated that a diet containing broccoli sprouts high in glucoraphanin (Grn), precursor of the phase 2 protein inducer sulforaphane, decreased oxidative stress and associated problems in male spontaneously hypertensive stroke-prone rats (SHRsp). We hypothesized that placing females on a Grn-containing (Grn+) diet would have similar positive effects and that the adult offspring of such females would have lower blood pressures and less tissue inflammation than offspring from mothers on control diet. We demonstrate that female SHRsp on a Grn+ diet had decreased oxidative stress and associated problems such as hypertension than females on control diet. The offspring of females on Grn+ diet also had lower blood pressures and less tissue inflammation in adulthood regardless of diet, with offspring placed on a Grn+ diet having the best health outcomes. We conclude that reducing oxidative stress in pregnant females has profound outcomes in the health of their adult offspring.


Impact of copper exposure on Pseudo-nitzschia spp. physiology and domoic acid production.
Microalgae have differing sensitivities to copper toxicity. Some species within the genus Pseudo-nitzschia produce domoic acid (DA), a phycotoxin that has been hypothesised to chelate Cu and ameliorate Cu toxicity to the cells. To better characterise the effect of Cu on Pseudo-nitzschia, a toxic strain of P. multiseries and a non-toxic strain of P. delicatissima were exposed to Cu(II) for 96 h (50 μg l(-1) for P. delicatissima and 50, 100 and 150 μg l(-1) for P. multiseries). Physiological measurements were performed daily on Pseudo-nitzschia cells using fluorescent probes and flow cytometry to determine the cell density, lipid concentration, chlorophyll autofluorescence, esterase activity, percentage of dead algal cells, and number of living and dead bacteria. Photosynthetic efficiency and O(2) consumption and production of cells were also measured using pulse amplitude modulated fluorometry and SDR Oxygen Sensor dish. The DA content was measured using ELISA kits. After 48 h of Cu exposure, P. delicatissima mortality increased dramatically whereas P. multiseries survival was unchanged (in comparison to control cells). Cellular esterase activity, chlorophyll autofluorescence, and lipid content significantly increased upon Cu exposure in comparison to control cells (24h for P. delicatissima, up to 96 h for P. multiseries). Bacterial concentrations in P. multiseries decreased significantly when exposed to Cu, whereas bacterial concentrations were similar between control and exposed populations of P. delicatissima. DA concentrations in P. multiseries were not modified by Cu exposure. Addition of DA to non-toxic P. delicatissima did not enhance cells survival; hence, extracellular DA does not protect Pseudo-nitzschia spp. against copper toxicity. Results suggested that cells of P. delicatissima are much more sensitive to Cu than P. multiseries. This difference is probably not related to the ability of P. multiseries to produce DA but could be explained by species differences in copper sensitivity, or a difference of bacterial community between the algal species.


Quantification of vitamin C in the rat brain in vivo using short echo-time 1H MRS.
Both ultrashort echo-time STEAM and MEGA-PRESS-edited spectroscopy were used to validate noninvasive quantification of vitamin C (ascorbate) in the developing rat brain, where changes in ascorbate concentration have been reported. Despite strong overlap with resonances from glutamine, glutamate, glutathione, and macromolecules, reliable quantification of ascorbate (Cramer-Rao lower bounds<0.2 micromol/g) by LCModel analysis of STEAM (TE=2 ms) spectra was possible at 9.4 T. Ascorbate concentrations quantified from the STEAM spectra were in very good agreement with concentrations calculated from fully resolved ascorbate resonances in MEGA-PRESS-edited spectra measured from identical volumes of interest. Ascorbate concentrations measured using STEAM decreased with increasing postnatal rat age, in agreement with published brain ascorbate concentrations measured in vitro using high-performance liquid chromatography (HPLC).