Enhanced Burst-Suppression and Disruption of Local Field Potential Synchrony in a Mouse Model of Focal Cortical Dysplasia Exhibiting Spike-Wave Seizures.
Focal cortical dysplasias (FCDs) are a common cause of brain seizures and are often associated with intractable epilepsy. Here we evaluated aberrant brain neurophysiology in an in vivo mouse model of FCD induced by neonatal freeze lesions (FLs) to the right cortical hemisphere (near S1). Linear multi-electrode arrays were used to record extracellular potentials from cortical and subcortical brain regions near the FL in anesthetized mice (5-13 months old) followed by 24 h cortical electroencephalogram (EEG) recordings. Results indicated that FL animals exhibit a high prevalence of spontaneous spike-wave discharges (SWDs), predominately during sleep (EEG), and an increase in the incidence of hyper-excitable burst/suppression activity under general anesthesia (extracellular recordings, 0.5%-3.0% isoflurane). Brief periods of burst activity in the local field potential (LFP) typically presented as an arrhythmic pattern of increased theta-alpha spectral peaks (4-12 Hz) on a background of low-amplitude delta activity (1-4 Hz), were associated with an increase in spontaneous spiking of cortical neurons, and were highly synchronized in control animals across recording sites in both cortical and subcortical layers (average cross-correlation values ranging from +0.73 to +1.0) with minimal phase shift between electrodes. However, in FL animals, cortical vs. subcortical burst activity was strongly out of phase with significantly lower cross-correlation values compared to controls (average values of -0.1 to +0.5, P < 0.05 between groups). In particular, a marked reduction in the level of synchronous burst activity was observed, the closer the recording electrodes were to the malformation (Pearson's Correlation = 0.525, P < 0.05). In a subset of FL animals (3/9), burst activity also included a spike or spike-wave pattern similar to the SWDs observed in unanesthetized animals. In summary, neonatal FLs increased the hyperexcitable pattern of burst activity induced by anesthesia and disrupted field potential synchrony between cortical and subcortical brain regions near the site of the cortical malformation. Monitoring the altered electrophysiology of burst activity under general anesthesia with multi-dimensional micro-electrode arrays may serve to define distinct neurophysiological biomarkers of epileptogenesis in human brain and improve techniques for surgical resection of epileptogenic malformed brain tissue.


Implementation of a Screening Program for Patients at Risk for Posttraumatic Stress Disorder.
Implantable cardioverter defibrillator (ICD) recipients who suffer from posttraumatic stress disorder (PTSD) are known to be associated with significant cardiac-specific mortality. Clinical observations suggest that PTSD is frequently undetected in ICD recipients followed up at electrophysiology (EP) outpatient clinics. Early recognition of PTSD is important to reduce the risk of serious manifestations on patient outcomes. All ICD recipients aged 19 years or older at the Washington University School of Medicine (WASHU) EP clinic, a large urban EP clinic, were invited to participate in the project. An informed consent letter with an attached primary care: posttraumatic stress disorder (PC: PTSD) survey was offered to the participants who met the inclusion criteria. Those who completed the survey were included in the project. Individuals with positive survey result were offered a referral to mental health services. Comparisons between PTSD and non-PTSD patients were done using a two-sample t-test for continuous variables. Using Fisher's exact test, PTSD prevalence was compared to the study by Ladwig et al in which prevalence was determined as the proportion of patients with positive findings of PTSD (n = 38/147). All analyses were conducted using SAS v9.4. The proportion of patients having PTSD was determined and an exact 95% confidence interval was evaluated based on the binomial distribution. Using a convenience sample, 50 ICD recipients (33 males and 17 females) were enrolled. The project had a 30-day outcome period. Nine (18%) of the 50 participants had positive PC: PTSD findings and all these nine participants were referred to a mental health specialist. The current project demonstrated an 18% (9/50) PTSD prevalence rate when compared to a 26% (38/147) prevalence rate in the study by Ladwig et al (P = 0.34). Although this project did not demonstrate 20% PTSD prevalence rate, as hypothesized, the 18% PTSD prevalence rate is consistent with previous research. The prevalence of PTSD noted in the current project is consistent with previous research and validates underrecognition of PTSD in ICD patients. Offering a referral to all ICD recipients at EP clinic visits with a positive PC: PTSD screening to a mental health specialist is an important step in reducing the risk of serious manifestations on patient outcomes.


Hypertension in hypoglycemic diabetics treated with beta-adrenergic antagonists.
To evaluate the response during insulin-induced hypoglycemia, diabetics treated with relatively selective or nonselective beta-adrenergic blocking agents were studied. Placebo, atenolol (100 mg/day), and propranolol (80 mg b.i.d.) were administered to 12 insulin-treated diabetics for 1 week in a double-blind, randomized, crossover fashion with a 2-week washout between treatments. Sufficient intravenous insulin (0.2-0.6 units/kg) was administered to decrease plasma glucose 68% from the basal level or to less than 60 mg/dl within 90 minutes. Blood pressure changes at the nadir of plasma glucose were +15.2/-9.9 mm Hg for placebo, +27.9/0 mm Hg for atenolol, and +38.8/+14.3 mm Hg for propranolol. Diastolic blood pressure changes induced by propranolol were significantly different from those induced by atenolol (p less than 0.01) and placebo (p less than 0.01), and systolic pressure changes were significantly different (p less than 0.02) between propranolol and atenolol. Mild seizures developed in two patients treated with propranolol. Their blood pressure changes at the plasma glucose nadir were +56/+22 and +86/+31 mm Hg. Other symptoms of hypoglycemia were more frequent during beta-blocker than during placebo treatment. Differences in response may be related to the relatively selective adrenergic blocking effect of these drugs.


Optimal charging profiles for mechanically constrained lithium-ion batteries.
The cost and safety related issues of lithium-ion batteries require intelligent charging profiles that can efficiently utilize the battery. This paper illustrates the application of dynamic optimization in obtaining the optimal current profile for charging a lithium-ion battery using a single-particle model while incorporating intercalation-induced stress generation. In this paper, we focus on the problem of maximizing the charge stored in a given time while restricting the development of stresses inside the particle. Conventional charging profiles for lithium-ion batteries (e.g., constant current followed by constant voltage) were not derived by considering capacity fade mechanisms. These charging profiles are not only inefficient in terms of lifetime usage of the batteries but are also slower since they do not exploit the changing dynamics of the system. Dynamic optimization based approaches have been used to derive optimal charging and discharging profiles with different objective functions. The progress made in understanding the capacity fade mechanisms has paved the way for inclusion of that knowledge in deriving optimal controls. While past efforts included thermal constraints, this paper for the first time presents strategies for optimally charging batteries by guaranteeing minimal mechanical damage to the electrode particles during intercalation. In addition, an executable form of the code has been developed and provided. This code can be used to identify optimal charging profiles for any material and design parameters.


The Ccr4-Not deadenylase complex constitutes the main poly(A) removal activity in C. elegans.
Post-transcriptional regulatory mechanisms are widely used to control gene expression programs of tissue development and physiology. Controlled 3' poly(A) tail-length changes of mRNAs provide a mechanistic basis of such regulation, affecting mRNA stability and translational competence. Deadenylases are a conserved class of enzymes that facilitate poly(A) tail removal, and their biochemical activities have been mainly studied in the context of single-cell systems. Little is known about the different deadenylases and their biological role in multicellular organisms. In this study, we identify and characterize all known deadenylases of Caenorhabditis elegans, and identify the germ line as tissue that depends strongly on deadenylase activity. Most deadenylases are required for hermaphrodite fertility, albeit to different degrees. Whereas ccr-4 and ccf-1 deadenylases promote germline function under physiological conditions, panl-2 and parn-1 deadenylases are only required under heat-stress conditions. We also show that the Ccr4-Not core complex in nematodes is composed of the two catalytic subunits CCR-4 and CCF-1 and the structural subunit NTL-1, which we find to regulate the stability of CCF-1. Using bulk poly(A) tail measurements with nucleotide resolution, we detect strong deadenylation defects of mRNAs at the global level only in the absence of ccr-4, ccf-1 and ntl-1, but not of panl-2, parn-1 and parn-2. Taken together, this study suggests that the Ccr4-Not complex is the main deadenylase complex in C. elegans germ cells. On the basis of this and as a result of evidence in flies, we propose that the conserved Ccr4-Not complex is an essential component in post-transcriptional regulatory networks promoting animal reproduction.


Konjac-Mannan and American ginsing: emerging alternative therapies for type 2 diabetes mellitus.
Despite significant achievements in treatment modalities and preventive measures, the prevalence of diabetes has risen exponentially in the last decade. Because of these limitations there is a continued need for new and more effective therapies. An increasing number of people are using dietary and herbal supplements, even though there is a general lack of evidence for their safety and efficacy. Consequently, science based medical and government regulators are calling for more randomized clinical studies to provide evidence of efficacy and safety. Our research group has selected two such promising and functionally complementary therapies for further investigation as potentially emerging alternative therapies for type 2 diabetes: Konjac-mannan (KJM) and American ginseng (AG). We have generated a mounting body of evidence to support the claim that rheologically-selected, highly-viscous KJM, and AG with a specific composition may be useful in improving diabetes control, reducing associated risk factors such as hyperlipidemia and hypertension, and ameliorating insulin resistance. KJM has a demonstrated ability to modulate the rate of absorption of nutrients from the small bowel, whereas AG has post-absorptive effects. Consequently, it appears that KJM and AG are acting through different, yet complementary, mechanisms: KJM by increasing insulin sensitivity and AG likely by enhancing insulin secretion. Before the therapeutic potential of KJM and AG as novel prandial agents for treatment of diabetes can be fully realized, further controlled trials with larger sample sizes and of longer duration are required. A determination of the active ingredients in AG, and the rheology-biology relationship of KJM are also warranted.


A short-term study of the effects of SBHAN, a novel compound, on gingival inflammation in the beagle dog.
Unique hydroxyl ion-modulating compounds based on the amino acid glycine have been developed that possess both antimicrobial and pro-healing properties. The purpose of the present study was to determine the effects of one of these compounds, 8.5% (w/v) sodium N, N-bis-2 (hydroxylethyl) aminoacetate (SBHA) with 0.3% (w/v) NaOH (SBHAN) on ligature-induced gingival inflammation in the beagle dog. Fifteen purebred beagle dogs were subjected to a 14-day oral hygiene regimen, consisting of manual scaling and daily toothbrushing with plain pumice. Gingival inflammation was then initiated by tying ligatures around 12 study teeth per dog and by placing the dogs on water-softened dog chow. After 30 days, ligatures were removed, dogs were placed on a hard diet and randomly assigned to five treatment groups by the flip of two coins. The five treatments included: 1) distilled, pyrogen-free water; 2) 8.5% (w/v) SBHAN; 3) 4.3% (w/v) SBHAN; 4) 0.12% chlorhexidine; and 5) 8.5% SBHA (w/v) (SBHAN without added NaOH). Solutions were placed in opaque spray bottles to shield their identity from the examiner. Treatment consisted of a daily aerosol application of 2 ml of each solution in a calibrated spray bottle to the affected teeth. The following measures were taken from the dogs at baseline (after hygienic phase), 30 days after initiation of gingival inflammation (before ligature removal), and 2 weeks and 4 weeks after ligature removal: 1) plaque index (PI); 2) gingival index (GI); 3) probing depths (PD); 4) relative attachment levels (RAL); and 5) gingival crevicular fluid volume (GCF). Analysis of subgingival plaque for anaerobic and aerobic colony forming units/ml was also performed at each time point. Gingival biopsies were performed, sectioned and stained with hematoxylin and eosin to quantify the inflammatory cell infiltrate (ICI). After ligature placement, increases were observed in PI, GI, PD, RAL, GCF, aerobic and anaerobic subgingival microbial counts, and ICI. After ligature removal, spontaneous resolution of gingival inflammation and plaque accumulation around the teeth of all dogs was observed with any treatment. Statistical analysis (Tukey's pairwise comparisons) of the mean PI, GI, PD, RAL, ICI, and GCF after 4 weeks of treatment with each agent, however, revealed that 8.5% SBHAN was significantly (P < 0.05) more effective than water, 4.3% SBHAN, or 8.5% SBHA in reducing PI, GI, PD, and GCF, but not RAL or ICI. Moreover, 0.12% chlorhexidine was more effective than water, 4.3% SBHAN, or 8.5% SBHA at reducing GI, PD, and GCF, but not PI, RAL, or ICI. No adverse reactions to the SBHAN were observed visually or histologically in any of the dogs during the course of the investigation. These data suggest that further investigation in a larger study population of the potential of SBHAN as an anti-gingivitis compound is warranted.


Temporalis function in anthropoids and strepsirrhines: an EMG study.
The major purpose of this study is to analyze anterior and posterior temporalis muscle force recruitment and firing patterns in various anthropoid and strepsirrhine primates. There are two specific goals for this project. First, we test the hypothesis that in addition to transversely directed muscle force, the evolution of symphyseal fusion in primates may also be linked to vertically directed balancing-side muscle force during chewing (Hylander et al. [2000] Am. J. Phys. Anthropol. 112:469-492). Second, we test the hypothesis of whether strepsirrhines retain the hypothesized primitive mammalian condition for the firing of the anterior temporalis, whereas anthropoids have the derived condition (Weijs [1994] Biomechanics of Feeding in Vertebrates; Berlin: Springer-Verlag, p. 282-320). Electromyographic (EMG) activities of the left and right anterior and posterior temporalis muscles were recorded and analyzed in baboons, macaques, owl monkeys, thick-tailed galagos, and ring-tailed lemurs. In addition, as we used the working-side superficial masseter as a reference muscle, we also recorded and analyzed EMG activity of the left and right superficial masseter in these primates. The data for the anterior temporalis provided no support for the hypothesis that symphyseal fusion in primates is linked to vertically directed jaw muscle forces during mastication. Thus, symphyseal fusion in primates is most likely mainly linked to the timing and recruitment of transversely directed forces from the balancing-side deep masseter (Hylander et al. [2000] Am. J. Phys. Anthropol. 112:469-492). In addition, our data demonstrate that the firing patterns for the working- and balancing-side anterior temporalis muscles are near identical in both strepsirrhines and anthropoids. Their working- and balancing-side anterior temporalis muscles fire asynchronously and reach peak activity during the power stroke. Similarly, their working- and balancing-side posterior temporalis muscles also fire asynchronously and reach peak activity during the power stroke. Compared to these strepsirrhines, however, the balancing-side posterior temporalis of anthropoids appears to have a relatively delayed firing pattern. Moreover, based on their smaller W/B ratios, anthropoids demonstrate a relative increase in muscle-force recruitment of the balancing-side posterior temporalis. This in turn suggests that anthropoids may emphasize the duration and magnitude of the power stroke during mastication. This hypothesis, however, requires additional testing. Furthermore, during the latter portion of the power stroke, the late activity of the balancing-side posterior temporalis of anthropoids apparently assists the balancing-side deep masseter in driving the working-side molars through the terminal portion of occlusion.


Evidence of a key role for photosynthetic oxygen release in oil storage in developing soybean seeds.
Based on the topographical analysis of photosynthesis and oil storage, we propose in a companion paper that photosynthetic oxygen release plays a key role in the local energy state, storage metabolism and flux toward lipid biosynthesis in developing soybean seeds. To test this hypothesis, we combined topographical analysis of ATP gradients across tissues, microsensor quantifications of internal O2 levels, assays of energy balance, metabolite profiles and isotope-labelling studies. Seeds show a marked degree of oxygen starvation in vivo (minimum O2 levels 0.1 kPa, approximately 1.3 microm), affecting ATP gradients, overall energy state, metabolite pools and storage activity. Despite the low light availability, photosynthesis supplies significant amounts of oxygen to the hypoxic seed tissue. This is followed by an increase in local ATP levels, most prominently within the lipid-synthesizing (inner) regions of the embryo. Concomitantly, partitioning of 14C-sucrose to lipids is increased, suggesting higher rates of lipid biosynthesis. It is concluded that both respiratory and biosynthetic fluxes are dynamically adjusted to photosynthetic oxygen supply.


Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
A method has been devised for the electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. The method results in quantitative transfer of ribosomal proteins from gels containing urea. For sodium dodecyl sulfate gels, the original band pattern was obtained with no loss of resolution, but the transfer was not quantitative. The method allows detection of proteins by autoradiography and is simpler than conventional procedures. The immobilized proteins were detectable by immunological procedures. All additional binding capacity on the nitrocellulose was blocked with excess protein; then a specific antibody was bound and, finally, a second antibody directed against the first antibody. The second antibody was either radioactively labeled or conjugated to fluorescein or to peroxidase. The specific protein was then detected by either autoradiography, under UV light, or by the peroxidase reaction product, respectively. In the latter case, as little as 100 pg of protein was clearly detectable. It is anticipated that the procedure will be applicable to analysis of a wide variety of proteins with specific reactions or ligands.


Rapid Assessment of Price Instability and Paucity of Medicines and Protection for COVID-19 Across Asia: Findings and Public Health Implications for the Future.
Background: Countries have introduced a variety of measures to prevent and treat COVID-19 with medicines and personal protective equipment (PPE), with some countries adopting preventative strategies earlier than others. However, there has been considerable controversy surrounding some treatments. This includes hydroxychloroquine where the initial hype and misinformation lead to shortages, price rises and suicides. Price rises and shortages have also been seen for PPE. Such activities can have catastrophic effects on patients where there are high co-payment levels and issues of affordability. Consequently, there is a need to investigate this further. Objective: Assess changes in the availability, utilization and prices of relevant medicines and PPE during the pandemic among a range of Asian countries. Our approach: Narrative literature review combined with interviews among community pharmacists to assess changes in consumption, prices and shortages of medicines and PPE from the beginning of March 2020 until end of May 2020. In addition, suggestions on ways to reduce misinformation. Results: 308 pharmacists took part from five Asian countries. There was an appreciable increase in the utilization of antimicrobials in Pakistan (in over 88% of pharmacies), with lower increases or no change in Bangladesh, India, Malaysia and Vietnam. Encouragingly, there was increased use of vitamins/immune boosters and PPE across the countries, as well as limited price rises for antimicrobials in India, Malaysia and Vietnam, although greater price rises seen for analgesics and vitamin C/immune boosters. Appreciable price increases were also seen for PPE across some countries. Conclusion: Encouraging to see increases in utilization of vitamins/immune boosters and PPE. However, increases in the utilization and prices of antimicrobials is a concern that needs addressing alongside misinformation and any unintended consequences from the pandemic. Community pharmacists can play a key role in providing evidence-based advice, helping to moderate prices, as well as helping address some of the unintended consequences of the pandemic.


Coercion in psychiatric care: what have we learned from research?
The use of coercion to assure that people with a mental illness receive treatment has been the focus one of the longest running controversies among mental health professionals. Until quite recently, however, this debate has been almost entirely based on abstract principles. Empirical research concerning coercion was quite limited. Recently, however, research in this field has blossomed. The development of a validated measure of perceived coercion has spawned a variety of new studies. A five-nation study in Scandinavia has begun the difficult task of assessing the impact of different legal systems and systems of care on perceived coercion. Two new studies have used random assignment designs to study the impact of outpatient commitment. This article reviews these and other studies and describes what they do, and do not, tell us about coercion in mental health treatment.


Two-tier charging strategies in public hospitals: implications for intra-hospital resource allocation and equity of access to hospital services.
Two-tier charging, the practice of offering separate qualities of service at different prices, is a growing practice in public hospitals internationally. This paper models two-tier charging as a Stackelberg game in which the Ministry of Health leads by setting prices and a representative hospital follows by setting quality levels to maximise surplus in response. Whether or not two-tier charging will secure cross-subsidy from superior to basic service users depends on the own and cross-quality effects of the demand functions for the two services. Under a range of assumptions, the policy will evoke cross-subsidy from basic to superior services.


Enabling quantitative robot-assisted compressional elastography via the extended Kalman filter.
Compressional or quasi-static elastography has demonstrated the capability to detect occult cancers in a variety of tissue types, however it has a serious limitation in that the resulting elastograms are generally qualitative whereas other forms of elastography, such as shear-wave, can produce absolute measures of elasticity for histopathological classification. We address this limitation by introducing a stochastic method using an extended Kalman filter and robot-assistance to obtain quantitative elastograms which are resilient to measurement noise and system uncertainty. In this paper, the probabilistic framework is described, which utilizes many ultrasound acquisitions obtained from multiple palpations, to fuse data and uncertainty from a robotic manipulator's joint encoders and force/torque sensor directly into the inverse reconstruction of the elastogram. Quantitative results are demonstrated over homogeneous and inclusion gelatin phantoms using a seven degree of freedom manipulator for a range of initial elasticity assumptions. Results imply resilience to poorly assumed initial conditions as all trials were within 5 kPa of the elasticity measured by a mechanical testing system. Moreover, the presence or absence of an inclusion is clear in all reconstructed elastograms even when artifacts are present in displacement fields, indicating further robustness to measurement noise. The proposed stochastic method allows fusion of data from a robot's sensors directly into compressional elastography image reconstruction which may stabilize optimization and improve accuracy. This approach provides a mathematical framework to readily incorporate measurements from additional sensors in future applications which may extend the capabilities of compressional elastography beyond that of producing quantitative elasticity measurements.


Crystallization and preliminary X-ray analysis of the core particle of bluetongue virus.
Core particles of bluetongue virus serotype 1 (South Africa) have been crystallized. The crystals, which grow up to 0.8 mm in diameter, belong to a primitive orthorhombic space group and have point group symmetry 222. The unit cell dimensions are 754 x 796 x 823 A3 and the crystallographic asymmetric unit contains one-half of a core particle. The best crystals diffract strongly to 4.8 A Bragg spacings, which is the maximum resolution to which we can measure data with the detectors available, suggesting that useful diffraction extends well beyond this. Core particles of serotype 10 have also been crystallized but the crystals have yet to be analyzed by X-ray diffraction.


Cortical gene transcription response patterns to water maze training in aged mice.
The hippocampus mediates the acquisition of spatial memory, but the memory trace is eventually transferred to the cortex. We have investigated transcriptional activation of pathways related to cognitive function in the cortex of the aged mouse by analyzing gene expression following water maze training. We identified genes that were differentially responsive in aged mice with accurate spatial performance during probe trials or repeated swimming sessions, relative to home cage conditions. Effective learners exhibited significantly greater activation of several pathways, such as the mitogen-activated protein kinase and insulin receptor signaling pathways, relative to swimmers. The genes encoding activity-related cytoskeletal protein (Arc) and brain-derived neurotrophic factor (BDNF) were upregulated in proficient learners, relative to swimmers and home cage controls, while the gene encoding Rho GTPase activating protein 32 (GRIT) was downregulated. We explored the regulation of Arc, BDNF, and GRIT expression in greater morphological detail using in situ hybridization. Recall during probe trials enhanced Arc expression across multiple cortical regions involved in the cognitive component of water maze learning, while BDNF expression was more homogeneously upregulated across cortical regions involved in the associational and sensorimotor aspects of water maze training. In contrast, levels of GRIT expression were uniformly reduced across all cortical regions examined. These results suggest that cortical gene transcription is responsive to learning in aged mice that exhibit behavioral proficiency, and support a distributed hypothesis of memory storage across multiple cortical compartments.


A cost-effectiveness analysis of three different methods of psychiatric case finding in the general population.
A study was carried out to compare the sensitivity as well as cost of three different methods of psychiatric case detection. It was found that the simplest method, which involved interviewing about 3% of the adult population, with a questionnaire taking only five minutes to complete, picked up as many adult epileptics and nearly as many psychotics as the inquiry with all the adults in the population using a sophisticated structured interview schedule. A method of medium complexity, in which the short five-minute questionnaire was given to one adult member of each family, detected in addition to all adult epileptics and psychotics, and many juvenile epileptics and mentally retarded. The cost of the simplest method was one-ninth and that of the method of medium complexity, one-fifth of the cost of the most sophisticated method. The method of medium complexity is recommended for use in the rural psychiatry programme of the developing countries.


Determinants of Default from Tuberculosis Treatment among Patients with Drug-Susceptible Tuberculosis in Karachi, Pakistan: A Mixed Methods Study.
Non-adherence to tuberculosis therapy can lead to drug resistance, prolonged infectiousness, and death; therefore, understanding what causes treatment default is important. Pakistan has one of the highest burdens of tuberculosis in the world, yet there have been no qualitative studies in Pakistan that have specifically examined why default occurs. We conducted a mixed methods study at a tuberculosis clinic in Karachi to understand why patients with drug-susceptible tuberculosis default from treatment, and to identify factors associated with default. Patients attending this clinic pick up medications weekly and undergo family-supported directly observed therapy. In-depth interviews were administered to 21 patients who had defaulted. We also compared patients who defaulted with those who were cured, had completed, or had failed treatment in 2013. Qualitative analyses showed the most common reasons for default were the financial burden of treatment, and medication side effects and beliefs. The influence of finances on other causes of default was also prominent, as was concern about the effect of treatment on family members. In quantitative analysis, of 2120 patients, 301 (14.2%) defaulted. Univariate analysis found that male gender (OR: 1.34, 95% CI: 1.04-1.71), being 35-59 years of age (OR: 1.54, 95% CI: 1.14-2.08), or being 60 years of age or older (OR: 1.84, 95% CI: 1.17-2.88) were associated with default. After adjusting for gender, disease site, and patient category, being 35-59 years of age (aOR: 1.49, 95% CI: 1.10-2.03) or 60 years of age or older (aOR: 1.76, 95% CI: 1.12-2.77) were associated with default. In multivariate analysis age was the only variable associated with default. This lack of identifiable risk factors and our qualitative findings imply that default is complex and often due to extrinsic and medication-related factors. More tolerable medications, improved side effect management, and innovative cost-reduction measures are needed to reduce default from tuberculosis treatment.


Genetic structure in contemporary south Tyrolean isolated populations revealed by analysis of Y-chromosome, mtDNA, and Alu polymorphisms.
Most of the inhabitants of South Tyrol in the eastern Italian Alps can be considered isolated populations because of their physical separation by mountain barriers and their sociocultural heritage. We analyzed the genetic structure of South Tyrolean populations using three types of genetic markers: Y-chromosome, mitochondrial DNA (mtDNA), and autosomal Alu markers. Using random samples taken from the populations of Val Venosta, Val Pusteria, Val Isarco, Val Badia, and Val Gardena, we calculated genetic diversity within and among the populations. Microsatellite diversity and unique event polymorphism diversity (on the Y chromosome) were substantially lower in the Ladin-speaking population of Val Badia compared to the neighboring German-speaking populations. In contrast, the genetic diversity of mtDNA haplotypes was lowest for the upper Val Venosta and Val Pusteria. These data suggest a low effective population size, or little admixture, for the gene pool of the Ladin-speaking population from Val Badia. Interestingly, this is more pronounced for Ladin males than for Ladin females. For the pattern of genetic Alu variation, both Ladin samples (Val Gardena and Val Badia) are among the samples with the lowest diversity. An admixture analysis of one German-speaking valley (Val Venosta) indicates a relatively high genetic contribution of Ladin origin. The reduced genetic diversity and a high genetic differentiation in the Rhaetoroman- and German-speaking South Tyrolean populations may constitute an important basis for future medical genetic research and gene mapping studies in South Tyrol.


Quantitative ultrastructure of Ia boutons in the ventral horn: scaling and positional relationships.
Three physiologically characterized spindle (group Ia) afferents were labeled by the intracellular injection of HRP and were processed for light-level reconstruction. Thirty-five boutons in the ventral horn were then selected for analysis. They were serially thin sectioned and characterized in terms of volume, total surface area and the surface area of apposition to postsynaptic neurons (apposed surface area), mitochondrial volume, vesicle and active zone features, relation to presynaptic contacts, postsynaptic profile size, and position within the terminal arbor. Virtually all of these characteristics were widely variable, both within the entire population and in the endings of a single fiber. Apposed surface area, mitochondrial volume, vesicle number, active zone vesicle number, active zone number, and total active zone area were highly correlated in a positive linear manner with bouton volume. This suggests a type of ultrastructural "size principle," in which the morphological features associated with synaptic release scale directly in proportion to bouton size. This pattern also extends to local circuit interactions: the extent of an Ia bouton's input from axoaxonal contacts (86% receive at least one axoaxonal contact) was directly proportional to its size. In addition, the characteristics of an Ia bouton were related to its position on the postsynaptic element and within the terminal arbor. Vesicle density, percentage mitochondrial volume, and active zone size increased as the postsynaptic process decreased in size, while volume, apposed surface area, active zone number and area, and vesicle number all decreased as one moved downstream within a terminal branch, with the exception of the terminal bouton. Vesicle density also decreased as one moved away from the dorsal root entry zone.


Inadequate stakeholder management and its effect on a coherent sinkhole risk management strategy: The case of the Merafong Local Municipality, South Africa.
The Merafong Local Municipality (MLM) has historically suffered financial and human losses because of the presence of dolomite and the consequent formation of sinkholes. There is a great need for the MLM to address the risk posed by sinkholes to ensure the continued safety of communities. However, as the risk is so pervasive, the MLM needs to coordinate their risk reduction strategies with a wide array of stakeholders in the municipality. Efficient stakeholder management is thus crucial if the sinkhole risk is to be addressed appropriately. This article reviews the current status of stakeholder management in the MLM as it pertains to the formulation of a holistic sinkhole risk reduction strategy. Findings indicate that there are serious deficiencies in the MLM's stakeholder management relating to key risk management processes such as community involvement in risk management structures, disaster risk assessment, training and awareness, and early warning and response. Improved stakeholder management could be characterised by the following factors: improved two-way communication between the municipality and community stakeholders, fostering a relationship based upon trust and equality amongst stakeholders, participation by a wide array of stakeholder groups affected by the sinkhole risk and a mutual commitment by all stakeholders to address the risk. These factors could contribute to enhancing current and future sinkhole risk reduction strategies.


The effectiveness of an intervention to reduce alcohol-related violence in premises licensed for the sale and on-site consumption of alcohol: a randomized controlled trial.
Premises licensed for the sale and consumption of alcohol can contribute to levels of assault-related injury through poor operational practices that, if addressed, could reduce violence. We tested the real-world effectiveness of an intervention designed to change premises operation, whether any intervention effect changed over time, and the effect of intervention dose. A parallel randomized controlled trial with the unit of allocation and outcomes measured at the level of individual premises. All premises (public houses, nightclubs or hotels with a public bar) in Wales, UK. A randomly selected subsample (n = 600) of eligible premises (that had one or more violent incidents recorded in police-recorded crime data; n = 837) were randomized into control and intervention groups. Intervention premises were audited by Environmental Health Practitioners who identified risks for violence and provided feedback by varying dose (informal, through written advice, follow-up visits) on how risks could be addressed. Control premises received usual practice. Police data were used to derive a binary variable describing whether, on each day premises were open, one or more violent incidents were evident over a 455-day period following randomization. Due to premises being unavailable at the time of intervention delivery 208 received the intervention and 245 were subject to usual practice in an intention-to-treat analysis. The intervention was associated with an increase in police recorded violence compared to normal practice (hazard ratio = 1.34, 95% confidence interval = 1.20-1.51). Exploratory analyses suggested that reduced violence was associated with greater intervention dose (follow-up visits). An Environmental Health Practitioner-led intervention in premises licensed for the sale and on-site consumption of alcohol resulted in an increase in police recorded violence.


Potassium-selective optical microsensors based on surface modified polystyrene microspheres.
Ion-selective microspheres based on surface modification of polystyrene particles (0.8 and 2.4 μm, diameter) are presented here for the first time. The traditional lipophilic sensing components of K(+) selective optodes (chromoionophore, ion-exchanger and ionophore) are adsorbed on the surface of the polystyrene particles using a simple mixed solvent method. The resulting microparticles respond to K(+) in an exhaustive sensing mode with excellent selectivity and rapid response time (t95% = 5 s).


Enantioseparation of phenothiazines through capillary electrophoresis with solid phase extraction and polymer based stacking.
This study developed a sensitive method involving capillary electrophoresis (CE) coupled with ultraviolet absorption for the simultaneous separation of chiral phenothiazine drugs at nanomolar concentration levels. The method consists of hydroxypropyl-γ-cyclodextrin (Hp-γ-CD) as a chiral selector and poly (diallyldimethylammonium chloride) (PDDAC)-based CE. Five pairs of d,l-phenothiazines were baseline separated using a background electrolyte containing 0.9% PDDAC, 5 mM Hp-γ-CD, and 100 mM tris(hydroxymethyl)aminomethane (Tris)-formate (pH 3.0). The five pairs were successfully stacked on the basis of the difference in viscosity between the PDDAC-containing background electrolyte and the sample solution, with almost no loss of resolution. The combination of a solid-phase extraction and PDDAC-mediated CE can efficiently improve the sensitivity of the phenothiazine enantiomers. Under optimal conditions, calibration graphs displayed the linear range between 6 and 1500 nM, with relative standard deviation values lower than 3.5% (n = 5). Detection limit ranged from 2.1 to 6.3 nM for target analytes, and 607- to 1555-fold enhancement was achieved. The practicality of using the proposed method to determine five pairs of d,l-phenothiazines in urine is also validated, in which recoveries between recoveries of all phenothiazines from urine ranged from 89% to 101%.


Effects of feed gas composition and catalyst thickness on carbon nanotube and nanofiber synthesis by plasma enhanced chemical vapor deposition.
Many engineering applications require carbon nanotubes with specific characteristics such as wall structure, chirality and alignment. However, precise control of nanotube properties grown to application specifications remains a significant challenge. Plasma-enhanced chemical vapor deposition (PECVD) offers a variety of advantages in the synthesis of carbon nanotubes in that several important synthesis parameters can be controlled independently. This paper reports an experimental study of the effects of reacting gas composition (percentage methane in hydrogen) and catalyst film thickness on carbon nanotube (CNT) growth and a computational study of gas-phase composition for the inlet conditions of experimentally observed carbon nanotube growth using different chemical reaction mechanisms. The simulations seek to explain the observed effects of reacting gas composition and to identify the precursors for CNT formation. The experimental results indicate that gas-phase composition significantly affects the synthesized material, which is shown to be randomly aligned nanotube and nanofiber mats for relatively methane-rich inlet gas mixtures and non-tubular carbon for methane-lean incoming mixtures. The simulation results suggest that inlet methane-hydrogen mixture coverts to an acetylene-methane-hydrogen mixture with minor amounts of ethylene, hydrogen atom, and methyl radical. Acetylene appears to be the indicator species for solid carbon formation. The simulations also show that inlet methane-hydrogen mixture does not produce enough gas-phase precursors needed to form quality CNTs below 5% CH4 concentrations in the inlet stream.


The WW domain of dystrophin requires EF-hands region to interact with beta-dystroglycan.
Skeletal muscle dystrophin is a 427 kDa protein thought to act as a link between the actin cytoskeleton and the extracellular matrix. Perturbations of the dystrophin-associated complex, for example, between dystrophin and the transmembrane glycoprotein beta-dystroglycan, may lead to muscular dystrophy. Previously, the cysteine-rich region and first half of the carboxy-terminal domain of dystrophin were shown to interact with beta-dystroglycan through a stretch of fifteen amino acids at the carboxy-terminus of beta-dystroglycan. This region of dystrophin implicated in binding beta-dystroglycan contains four modular protein domains: a WW domain, two putative Ca2+-binding EF-hand motifs, and a putative zinc finger ZZ domain. The WW domain is a globular domain of 38-40 amino acids with two highly conserved tryptophan residues spaced 20-22 amino acids apart. A subset of WW domains was shown to bind ligands that contain a Pro-Pro-x-Tyr core motif (where x is any amino acid). Here we elucidate the role of the WW domain of dystrophin and surrounding sequence in binding beta-dystroglycan. We show that the WW domain of dystrophin along with the EF-hand motifs binds to the carboxy-terminus of beta-dystroglycan. Through site-specific mutagenesis and in vitro binding assays, we demonstrate that binding of dystrophin to the carboxy-terminus of beta-dystroglycan occurs via a beta-dystroglycan Pro-Pro-x-Tyr core motif. Targeted mutagenesis of conserved WW domain residues reveals that the dystrophin/beta-dystroglycan interaction occurs primarily through the WW domain of dystrophin. Precise mapping of this interaction could aid in therapeutic design.


Inhibition of androgen receptor-mediated transcription by amino-terminal enhancer of split.
A yeast two-hybrid assay has identified an androgen-dependent interaction of androgen receptor (AR) with amino-terminal enhancer of split (AES), a member of the highly conserved Groucho/TLE family of corepressors. Full-length AR, as well as the N-terminal fragment of AR, showed direct interactions with AES in in vitro protein-protein interaction assays. AES specifically inhibited AR-mediated transcription in a well-defined cell-free transcription system and interacted specifically with the basal transcription factor (TFIIE) in HeLa nuclear extract. These observations implicate AES as a selective repressor of ligand-dependent AR-mediated transcription that acts by directly interacting with AR and by targeting the basal transcription machinery.


Perinatal Exposure to Low-Dose Bisphenol-A Disrupts the Structural and Functional Development of the Hypothalamic Feeding Circuitry.
Bisphenol-A (BPA) is a component of polycarbonate and other plastics to which humans are regularly exposed at low levels. BPA is characterized as an endocrine disruptor because of observations of its estrogenic activity in various experimental models. We have previously shown evidence of disrupted hypothalamic feeding circuitry and leptin sensitivity in adult BPA-exposed animals subjected to a high-fat diet, but because these animals were already exhibiting a diet-induced obese phenotype, we could not rule out the possibility that these observations were simply consequences of the obesity, not a preexisting phenotype produced by BPA exposure. Here, we studied leptin sensitivity and hypothalamic structure in young BPA-exposed animals before the onset of a body weight or metabolic phenotype. Pregnant and lactating CD-1 mice were exposed to either BPA or diethylstilbestrol (DES) at low, environmentally relevant doses via their diet. Studies of leptin function and neurobiology were conducted on offspring at several time points. Young adult offspring from this experiment were resistant to leptin-induced suppression of food intake, body weight loss, and hypothalamic pro-opiomelanocortin (POMC) upregulation. Both male and female BPA-exposed mice showed a reduced density of POMC projections into the paraventricular nucleus of the hypothalamus (PVN). BPA- and DES-exposed pups had respectively delayed and blunted postnatal leptin surges, and POMC projections into the PVN were rescued in female BPA-exposed animals given daily injections of supplemental leptin. Our findings suggest that BPA, a putative obesogen, may exert its effects through developmental programming of the hypothalamic melanocortin circuitry, permanently altering the neurobiology of metabolic homeostasis.


Snake venomics of Bothrops punctatus, a semiarboreal pitviper species from Antioquia, Colombia.
Bothrops punctatus is an endangered, semi-arboreal pitviper species distributed in Panamá, Colombia, and Ecuador, whose venom is poorly characterized. In the present work, the protein composition of this venom was profiled using the 'snake venomics' analytical strategy. Decomplexation of the crude venom by RP-HPLC and SDS-PAGE, followed by tandem mass spectrometry of tryptic digests, showed that it consists of proteins assigned to at least nine snake toxin families. Metalloproteinases are predominant in this secretion (41.5% of the total proteins), followed by C-type lectin/lectin-like proteins (16.7%), bradykinin-potentiating peptides (10.7%), phospholipases A2 (93%), serine proteinases (5.4%), disintegrins (38%), L-amino acid oxidases (3.1%), vascular endothelial growth factors (17%), and cysteine-rich secretory proteins (1.2%). Altogether, 6.6% of the proteins were not identified. In vitro, the venom exhibited proteolytic, phospholipase A2, and L-amino acid oxidase activities, as well as angiotensin-converting enzyme (ACE)-inhibitory activity, in agreement with the obtained proteomic profile. Cytotoxic activity on murine C2C12 myoblasts was negative, suggesting that the majority of venom phospholipases A2 likely belong to the acidic type, which often lack major toxic effects. The protein composition of B. punctatus venom shows a good correlation with toxic activities here and previously reported, and adds further data in support of the wide diversity of strategies that have evolved in snake venoms to subdue prey, as increasingly being revealed by proteomic analyses.


Hypoxia-preconditioned adipose tissue-derived mesenchymal stem cell increase the survival and gene expression of engineered neural stem cells in a spinal cord injury model.
Hypoxic preconditioning (HP) is a novel strategy to make stem cells resistant to the ischemic environment they encounter after transplantation into injured tissue; this strategy improves survival of both the transplanted cells and the host cells at the injury site. Using both in vitro and in vivo injury models, we confirmed that HP-treated adipose tissue-derived mesenchymal stem cells (HP-AT-MSCs) increased cell survival and enhanced the expression of marker genes in DsRed-engineered neural stem cells (NSCs-DsRed). Similar to untreated AT-MSCs, HP-AT-MSCs had normal morphology and were positive for the cell surface markers CD90, CD105, and CD29, but not CD31. In three in vitro ischemic-mimicking injury models, HP-AT-MSCs significantly increased both the viability of NSCs-DsRed and the expression of DsRed and clearly reduced the number of annexin-V-positive apoptotic NSCs-DsRed and the expression of the apoptotic factor Bax. Consistent with the in vitro assay, co-transplantation of NSCs-DsRed with HP-AT-MSCs significantly improved the survival of the NSCs-DsRed, resulting in an increased expression of the DsRed reporter gene at the transplantation site in a rat spinal cord injury (SCI) model. These findings suggest that the co-transplantation of HP-AT-MSCs with engineered NSCs can improve both the cell survival and the gene expression of the engineered NSCs, indicating that this novel strategy can be used to augment the therapeutic efficacy of combined stem cell and gene therapies for SCI.


Evaluation of immunohistochemistry in identifying Bartonella henselae in cat-scratch disease.
Cat-scratch disease (CSD) is largely due to infection with Bartonella henselae. Microbiologic detection is difficult, and molecular testing is not readily available. A monoclonal antibody (mAB) to B henselae has become commercially available. We evaluated the usefulness of immunohistochemical analysis (IHC) for diagnosing CSD on surgical specimens and compared these results with polymerase chain reaction (PCR) detection and serologic testing for B henselae. We studied 24 formalin-fixed, paraffin-embedded (FFPE) cases of lymphadenitis with histologic and/or clinical suspicion of CSD. Control cases included 14 cases of lymphadenopathy other than CSD. FFPE tissue sections were evaluated with an mAB to B henselae, Steiner silver stain (SSS), and PCR that targeted B henselae and Bartonella quintana. Positive cases were as follows: SSS, 11 (46%); PCR, 9 (38%); and IHC, 6 (25%). Only 2 cases (8%) were positive for all 3 studies. All control cases were negative for IHC and PCR. The diagnostic sensitivity of these 3 tests is low for CSD. SSS seems to be the most sensitive test but is the least specific. PCR is more sensitive than IHC and may, therefore, serve as a helpful second-line test on all IHC- cases.


Effect of piped water supply on human water contact patterns in a Schistosoma haematobium-endemic area in Coast Province, Kenya.
The effect of a piped water supply on human water contact in a Schistosoma haematobium-endemic area in Coast Province, Kenya was studied. After the construction of five community standpipes and one shower unit, there was a 35.1% reduction in the number of people observed using river water, a 44.1% reduction in the frequency of contact with river water, and a 25.4% reduction in the amount of contact. The frequency of river water contact per person also decreased significantly, but the amount of contact per person did not decrease. The total frequency of contact decreased significantly except for washing clothes by the river, washing utensils, and fishing. The frequency per person did not change for most of the activities and significantly increased for washing clothes. The frequency of river water contact in households with high piped water consumption showed a significant decrease compared with those with low piped water consumption. The volume of consumption of piped water was inversely proportional to the distance from the home to the community standpipe. These results indicate that in the study area, the effect of a piped water supply on river water contact behavior was heterologous while the total river water contact decreased significantly, and that the piped water had a beneficial effect on some villagers but very little effect on others.


Treatment of chronic tinnitus with repeated sessions of prefrontal transcranial direct current stimulation: outcomes from an open-label pilot study.
Tinnitus is the perception of sound in the absence of an environmental sound source. Abnormal activity in central auditory pathways is considered as the neuronal correlate of tinnitus. However, there is increasing evidence from neuroimaging studies for an additional involvement of the frontal cortex in the pathophysiology of tinnitus, especially concerning its attentional and emotional aspects. Recently, in a subgroup of tinnitus patients, temporary reduction of tinnitus intensity and tinnitus-related distress has been reported after bifrontal tDCS with the anode over the right and the cathode over the left dorsolateral prefrontal cortex (DLPFC). The aim of this study was to investigate whether repeated application of bifrontal tDCS results in longer-lasting reduction of tinnitus and may represent a potential treatment approach. Thirty-two patients with chronic and treatment-resistant tinnitus received six sessions of bifrontal tDCS (1.5 mA, 30 min, two sessions per week) with the anode over the right and the cathode over the left DLPFC. Treatment outcome was assessed with several standardized tinnitus questionnaires, numeric rating scales, and a depression scale. In the entire group, beneficial effects of bifrontal tDCS on tinnitus were found for numeric rating scores of loudness, unpleasantness, and discomfort, but not in tinnitus or depression scales. Exploratory analysis revealed an effect of gender on treatment effects with female patients demonstrating a better response in several scores. Our open-label pilot study suggests some beneficial effect of bifrontal tDCS (anode right and cathode left) in the treatment of severe tinnitus, warranting further controlled studies.


Quality of acute episodic care in investor-owned ambulatory health centers.
This article examines the quality of acute episodic care for five diagnostic categories amenable to one-visit diagnosis and treatment at the nation's largest chain of investor-owned ambulatory care centers. A total of 803 medical records were audited for five common conditions and measured against specific protocols. In four of the five diagnostic categories studied--pharyngitis, otitis media, vaginitis, and use of tetanus immunization--42-97% of patients received care that met or exceeded the standards set by a panel of practicing academic physicians. In follow-up of an incidental high blood pressure reading, however, study physicians met the standard only 24% of the time. Some overprescribing and overtreatment with immunizations were detected. As far as comparison is possible to other studies, results suggest that care in this setting falls within the range of experience that has been reported for other types of practices. In spite of direct economic incentives to increase volume, little evidence was found of overuse of ancillary tests or unnecessary scheduling of repeat visits.


Selenomethionine substitution of orotidine-5'-monophosphate decarboxylase causes a change in crystal contacts and space group.
Orotidine 5'-monophosphate decarboxylase (ODCase) catalyses the decarboxylation of orotidine 5'-monophosphate to uridine 5'-monophosphate, the last step in the de novo biosynthesis of uridine 5'-monophosphate. In order to determine the structure of ODCase from Escherichia coli by the multi-wavelength anomalous dispersion technique, both native and SeMet-substituted proteins have been produced and purified. During the production of SeMet ODCase, it was observed that SeMet was the only amino acid that it was necessary to add to the defined medium during expression. SeMet-substituted ODCase in complex with the inhibitor 1-(5'-phospho-beta-D-ribofuranosyl)barbituric acid crystallizes under similar conditions as the native enzyme. In contrast to the native enzyme, where the crystals belong to the orthorhombic space group P2(1)2(1)2(1), the SeMet-substituted enzyme crystallizes in the monoclinic space group P2(1), with a quadrupling of the volume of the asymmetric unit. Despite the drastic difference in symmetry, the overall crystal packing is effectively identical in the two crystal forms. The change in space group appears to originate in differences in the crystal contacts near the SeMet and Met residues. These differences can be rationalized in terms of SeMet's larger size and hydrophobicity.


Structural and functional insights into peptidoglycan access for the lytic amidase LytA of Streptococcus pneumoniae.
The cytosolic N-acetylmuramoyl-l-alanine amidase LytA protein of Streptococcus pneumoniae, which is released by bacterial lysis, associates with the cell wall via its choline-binding motif. During exponential growth, LytA accesses its peptidoglycan substrate to cause lysis only when nascent peptidoglycan synthesis is stalled by nutrient starvation or β-lactam antibiotics. Here we present three-dimensional structures of LytA and establish the requirements for substrate binding and catalytic activity. The solution structure of the full-length LytA dimer reveals a peculiar fold, with the choline-binding domains forming a rigid V-shaped scaffold and the relatively more flexible amidase domains attached in a trans position. The 1.05-Å crystal structure of the amidase domain reveals a prominent Y-shaped binding crevice composed of three contiguous subregions, with a zinc-containing active site localized at the bottom of the branch point. Site-directed mutagenesis was employed to identify catalytic residues and to investigate the relative impact of potential substrate-interacting residues lining the binding crevice for the lytic activity of LytA. In vitro activity assays using defined muropeptide substrates reveal that LytA utilizes a large substrate recognition interface and requires large muropeptide substrates with several connected saccharides that interact with all subregions of the binding crevice for catalysis. We hypothesize that the substrate requirements restrict LytA to the sites on the cell wall where nascent peptidoglycan synthesis occurs. Streptococcus pneumoniae is a human respiratory tract pathogen responsible for millions of deaths annually. Its major pneumococcal autolysin, LytA, is required for autolysis and fratricidal lysis and functions as a virulence factor that facilitates the spread of toxins and factors involved in immune evasion. LytA is also activated by penicillin and vancomycin and is responsible for the lysis induced by these antibiotics. The factors that regulate the lytic activity of LytA are unclear, but it was recently demonstrated that control is at the level of substrate recognition and that LytA required access to the nascent peptidoglycan. The present study was undertaken to structurally and functionally investigate LytA and its substrate-interacting interface and to determine the requirements for substrate recognition and catalysis. Our results reveal that the amidase domain comprises a complex substrate-binding crevice and needs to interact with a large-motif epitope of peptidoglycan for catalysis.


Leptin Restores Endothelial Function via Endothelial PPARγ-Nox1-Mediated Mechanisms in a Mouse Model of Congenital Generalized Lipodystrophy.
Leptin is the current treatment for metabolic disorders associated with acquired and congenital generalized lipodystrophy (CGL). Although excess leptin levels have been associated with vascular inflammation and cardiovascular disease in the context of obesity, the effects of chronic leptin treatment on vascular function remain unknown in CGL. Here, we hypothesized that leptin treatment will improve endothelial function via direct vascular mechanisms. We investigated the cardiovascular consequences of leptin deficiency and supplementation in male gBscl2-/- (Berardinelli-Seip 2 gene-deficient) mice-a mouse model of CGL. CGL mice exhibited reduced adipose mass and leptin levels, as well as impaired endothelium-dependent relaxation. Blood vessels from CGL mice had increased NADPH Oxidase 1 (Nox1) expression and reactive oxygen species production, and selective Nox1 inhibition restored endothelial function. Remarkably, chronic and acute leptin supplementation restored endothelial function via a PPARγ-dependent mechanism that decreased Nox1 expression and reactive oxygen species production. Selective ablation of leptin receptors in endothelial cells promoted endothelial dysfunction, which was restored by Nox1 inhibition. Lastly, we confirmed in aortic tissue from older patients undergoing cardiac bypass surgery that acute leptin can promote signaling in human blood vessels. In conclusion, in gBscl2-/- mice, leptin restores endothelial function via peroxisome proliferator activated receptor gamma-dependent decreases in Nox1. Furthermore, we provide the first evidence that vessels from aged patients remain leptin sensitive. These data reveal a new direct role of leptin receptors in the control of vascular homeostasis and present leptin as a potential therapy for the treatment of vascular disease associated with low leptin levels.


Prioritising pathogens for the management of severe febrile patients to improve clinical care in low- and middle-income countries.
Severe febrile illness without a known source (SFWS) is a challenge for clinicians when deciding how to manage a patient, particularly given the wide spectrum of potential aetiologies that contribute to fever. These infections are difficult to distinguish clinically, and accurate diagnosis requires a plethora of diagnostics including blood cultures, imaging techniques, molecular or serological tests, and more. When laboratory services are available, a limited test menu hinders clinical decision-making and antimicrobial stewardship, leading to empiric treatment and suboptimal patient outcomes. To specifically address SFWS, this work aimed to identify priority pathogens for a globally applicable panel for fever causing pathogens. A pragmatic two-pronged approach combining currently available scientific data in an analytical hierarchy process and systematically gathered expert input, was designed to address the lack of comprehensive global aetiology data. The expert re-ranked list was then further adapted for a specific use case to focus on community acquired infections in whole blood specimens. The resulting list was further analysed to address different geographical regions (Asia, Africa, and Latin America), and Cohen kappa scores of agreement were calculated. The expert ranked prioritized pathogen list generated as part of this two-pronged approach included typhoidal Salmonella, Plasmodium species and Mycobacterium tuberculosis as the top 3 pathogens. This pathogen list was then further adapted for the SFWS use case to develop a final pathogen list to inform product development. Subsequent analysis comparing the relevance of the SFWS pathogen list to multiple populations and geographical regions showed that the SFWS prioritized list had considerable utility across Africa and Asia, but less so for Latin America. In addition, the list showed high levels of agreement across different patient sub-populations, but lower relevance for neonates and symptomatic HIV patients. This work highlighted once again the challenges of prioritising in global health, but it also shows that taking a two-pronged approach, combining available prevalence data with expert input, can result in a broadly applicable priority list. This comprehensive utility is particularly important in the context of product development, where a sufficient market size is essential to achieve a sustainable commercialized diagnostic product to address SFWS.


Primary cilium formation requires von hippel-lindau gene function in renal-derived cells.
Biallelic inactivation of the von Hippel-Lindau tumor suppressor gene, VHL, occurs in the majority of renal clear cell carcinomas (RCC). VHL's function, regulating the degradation of hypoxia-inducible factor alpha (HIFalpha) subunits, explains the angiogenic nature of these tumors, but not tumor initiation. Because the development of renal cysts precedes tumor formation, and because the dysfunction of primary cilium is a common pathogenic mechanism in polycystic kidney diseases, we determined whether kidney-derived VHL- cells required VHL for the generation of cilium. Ectopic expression of VHL in RCC(VHL-) cells induced increased polarization and primary cilium formation. Cilium formation correlated directly with the expression of both wild-type VHL isoforms and a VHL mutant not associated with RCC development, whereas expression of RCC-associated VHL mutants did not support ciliogenesis. Requirement of VHL for ciliogenesis was independent of HIFalpha abundance. These data indicate separable independent functions for VHL (HIFalpha degradation and differentiation) and suggest a mechanism whereby disruption of both functions is required for renal carcinogenesis.


Identification and assessment of ecotoxicological hazards attributable to pollutants in urban wet weather discharges.
Urbanization has led to considerable pressure on urban/suburban aquatic ecosystems. Urban Wet Weather Discharges (UWWD) during rainfall events are a major source of pollutants leached onto and into urban surfaces and sewers, which in turn affect aquatic ecosystems. We assessed the ecotoxicity of the different compounds identified in UWWD and identified the hazard represented by each of them. To this end, hazard quotient (HQ) values were calculated for each compound detected in UWWD based on their predicted no effect concentration (PNEC) values and their maximum measured effluent concentrations (MECmax) found in the dissolved part of UWWD. For the 207 compounds identified in UWWD, sufficient data existed for 165 of them to calculate their PNEC. The ecotoxicity of these compounds varied greatly. Pesticides represented a high proportion of the wide variety of hazardous compounds whose HQ values were calculated (94 HQ values), and they were among the most hazardous pollutants (HQ > 1000) transported by stormwater. The hazard of combined sewer overflows (CSO) was linked mainly to heavy metals and pharmaceutical compounds. Consequently, the monitoring of these pollutants should be a priority in the future. The hazard level of certain pollutants could have been underestimated due to their adsorption onto particles, leading to their low concentration in the dissolved phase of UWWD. Hence, an in-depth study of these pollutants will be required to clarify their effects on aquatic organisms.


Estrogen receptor beta expression in prostate adenocarcinoma.
Prostate cancer is the most commonly diagnosed cancer in men and the second leading cause of cancer death in men. Estrogen induction of cell proliferation is a crucial step in carcinogenesis of gynecologic target tissues, and there are many studies recently done, showing that prostate cancer growth is also influenced by estrogen. The characterization of estrogen receptor beta (ER-b) brought new insight into the mechanisms underlying estrogen signalling. In the present study, we investigated the expression of estrogen receptor-b (ER-b) in human prostate cancer tissues. We selected 52 paraffin-embedded blocks of prostate needle biopsies in a cross-sectional study to determine frequency and rate of ER-b expression in different grades of prostate adenocarcinoma according to Gleason grading system. Immunohistochemical staining of tissue sections by monoclonal anti ER-b antibody was performed using an Envision method visualising system. ER-b expression was seen in tumoral cells of prostatic carcinoma in all 29 cases with low and intermediate tumors (100%) and 19 of 23 cases with high grade tumor (83%). Mean rate of ER-b expression in low & intermediate grade cancers was 68.41% (SD = 25.63) whereas high grade cancers showed 49.48% rate of expression (SD = 28.79). ER-b expression is reduced in high grade prostate cancers compared to low & intermediate grade ones (P value 0.027).


Subulura halli (Ascaridida: Subuluridae) from the endangered great bustard Otis tarda Linnaeus (Aves: Gruiformes) in China.
Subulurid nematodes identified as Subulura halli Barreto, 1918 were collected from the endangered bird Otis tarda Linnaeus (Gruiformes: Otididae) in China. A detailed redescription of the hitherto poorly known species is presented using both light and, for the first time, scanning electron microscopy. Previously unreported and erroneous morphological features of taxonomic significance are revealed. This species can be readily distinguished from its congeners by the relatively long oesophagus (1.47-1.92 mm long, representing 10.6-16.9% of body length), the number and arrangement of male caudal papillae (11 pairs in total, arranged as five pairs of precloacal and six pairs of postcloacal papillae), the equal length of spicules (1.35-1.52 mm long, representing 10.7-13.7% of body length) and the presence of a small medioventral, precloacal papilla in the male.


Disease-associated KBTBD4 mutations in medulloblastoma elicit neomorphic ubiquitylation activity to promote CoREST degradation.
Medulloblastoma is the most common malignant brain tumour in children. Genomic studies have identified distinct disease subgroups: wnt/wingless (WNT), sonic hedgehog (SHH), and non-WNT/non-SHH, comprising group 3 and group 4. Alterations in WNT and SHH signalling form the pathogenetic basis for their subgroups, whereas those for non-WNT/non-SHH tumours remain largely elusive. Recent analyses have revealed recurrent in-frame insertions in the E3 ubiquitin ligase adaptor Kelch Repeat and BTB Domain Containing 4 (KBTBD4) in cases of group 3/4 medulloblastoma. Critically, group 3/4 tumours with KBTBD4 mutations typically lack other gene-specific alterations, such as MYC amplification, indicating KBTBD4 insertion mutations as the primary genetic driver. Delineating the role of KBTBD4 mutations thus offers significant opportunities to understand tumour pathogenesis and to exploit the underpinning mechanisms therapeutically. Here, we show a novel mechanism in cancer pathogenesis whereby indel mutations in KBTBD4 drive its recognition of neo-substrates for degradation. We observe that KBTBD4 mutants promote the recruitment and ubiquitylation of the REST Corepressor (CoREST), which forms a complex to modulate chromatin accessibility and transcriptional programmes. The degradation of CoREST promoted by KBTBD4 mutation diverts epigenetic programmes inducing significant alterations in transcription to promote increased stemness of cancer cells. Transcriptional analysis of >200 human group 3 and 4 medulloblastomas by RNA-seq, highlights the presence of CoREST and stem-like signatures in tumours with KBTBD4 mutations, which extend to a further sub-set of non-mutant tumours, suggesting CoREST alterations as a novel pathogenetic mechanism of wide relevance in groups 3 and 4. Our findings uncover KBTBD4 mutation as a novel driver of epigenetic reprogramming in non-WNT/non-SHH medulloblastoma, establish a novel mode of tumorigenesis through gain-of-function mutations in ubiquitin ligases (neo-substrate recruitment) and identify both mutant KBTBD4 and CoREST complexes as new druggable targets for improved tumour-specific therapies.


Use of Intravenous Recombinant Tissue Plasminogen Activator in Patients With Acute Ischemic Stroke Who Take Non-Vitamin K Antagonist Oral Anticoagulants Before Stroke.
Intravenous rt-PA (recombinant tissue-type plasminogen activator) is effective in improving outcomes in ischemic stroke; however, there are few data on the use of rt-PA in patients who are receiving a non-vitamin K antagonist oral anticoagulant (NOAC). Using data from the American Heart Association Get With The Guidelines-Stroke Registry, we examined the outcomes of use of thrombolytic therapy in patients with ischemic stroke who received anticoagulation with NOACs versus those on warfarin (international normalized ratio <1.7) or not on anticoagulation from 1289 registry hospitals between October 2012 and March 2015. Of 42 887 patients with ischemic stroke treated with intravenous rt-PA within 4.5 hours, 251 were taking NOACs (dabigatran 87, rivaroxaban 129, and apixaban 35) before their stroke, 1500 were taking warfarin, and 41 136 were on neither. Patients on NOACs or warfarin were older, had more comorbid conditions, and experienced more severe strokes than did those who were not on anticoagulation (median National Institutes of Health Stroke Scale 12, 13, and 9, respectively). Unadjusted rates of symptomatic intracranial hemorrhage in the NOAC, warfarin, and none groups were 4.8%, 4.9%, and 3.9%, respectively (P=0.11). In comparison with those not on anticoagulation, the adjusted odds ratio for symptomatic intracranial hemorrhage for those on NOACs was 0.92 (95% confidence interval, 0.51-1.65) and for those on warfarin the adjusted odds ratio was 0.85 (95% confidence interval, 0.66-1.10). There were also no significant differences in the risk for life-threatening/serious systemic hemorrhage, any rt-PA complication, in-hospital mortality, and modified Rankin Scale at discharge across 3 groups. Similar results were also found after propensity score matching. Although experience of using rt-PA in patients with ischemic stroke on a NOAC is limited, these preliminary observations suggest that rt-PA appears to be reasonably well tolerated without prohibitive risks for adverse events among selected NOAC-treated patients. Future studies should evaluate the safety and efficacy of intravenous rt-PA in patients with ischemic stroke who are taking NOACs.


High Operation Stability of Ultraflexible Organic Solar Cells with Ultraviolet-Filtering Substrates.
Ultralightweight and flexible power sources are essential for driving textile or wearable electronic devices and soft robots because they do not induce discomfort or limit movement when they are attached to human skin, textiles, or soft actuators. Organic solar cells (OSCs) are good candidates for developing such power sources because they have the advantages of being lightweight and flexible. However, achieving operational stability and ultrathin shape simultaneously remains difficult because the ultrathin substrate cannot prevent the penetration of ultraviolet (UV) light, which is major a cause for the degradation of OSCs. Here, ultrathin OSCs that show great operational stability and high performance are reported. The 1.3 µm thick transparent polyimide utilized as a substrate can block light of 350 nm wavelength in the UV range by 90%. The ultrathin OSCs with the transparent polyimide substrate produce a power conversion efficiency (PCE) of 9.0% and realize both photostability and operational stability. The PCE was maintained at 90% after 3 h in a maximum power point tracking test, indicating much better operational stability than the reference rigid OSCs.


Inter-field variability in the microbial communities of hydrothermal vent deposits from a back-arc basin.
Diverse microbial communities thrive on and in deep-sea hydrothermal vent mineral deposits. However, our understanding of the inter-field variability in these communities is poor, as limited sampling and sequencing efforts have hampered most previous studies. To explore the inter-field variability in these communities, we used barcoded pyrosequencing of the variable region 4 (V4) of the 16S rRNA gene to characterize the archaeal and bacterial communities of over 30 hydrothermal deposit samples from six vent fields located along the Eastern Lau Spreading Center. Overall, the bacterial and archaeal communities of the Eastern Lau Spreading Center are similar to other active vent deposits, with a high diversity of Epsilonproteobacteria and thermophilic Archaea. However, the archaeal and bacterial communities from the southernmost vent field, Mariner, were significantly different from the other vent fields. At Mariner, the epsilonproteobacterial genus Nautilia and the archaeal family Thermococcaceae were prevalent in most samples, while Lebetimonas and Thermofilaceae were more abundant at the other vent fields. These differences appear to be influenced in part by the unique geochemistry of the Mariner fluids resulting from active degassing of a subsurface magma chamber. These results show that microbial communities associated with hydrothermal vent deposits in back-arc basins are taxonomically similar to those from mid-ocean ridge systems, but differences in geologic processes between vent fields in a back-arc basin can influence microbial community structure.


Chromatin regulatory mechanisms in pluripotency.
Stem cells of all types are characterized by a stable, heritable state permissive of multiple developmental pathways. The past five years have seen remarkable advances in understanding these heritable states and the ways that they are initiated or terminated. Transcription factors that bind directly to DNA and have sufficiency roles have been most easy to investigate and, perhaps for this reason, are most solidly implicated in pluripotency. In addition, large complexes of ATP-dependent chromatin-remodeling and histone-modification enzymes that have specialized functions have also been implicated by genetic studies in initiating and/or maintaining pluripotency or multipotency. Several of these ATP-dependent remodeling complexes play non-redundant roles, and the esBAF complex facilitates reprogramming of induced pluripotent stem cells. The recent finding that virtually all histone modifications can be rapidly reversed and are often highly dynamic has raised new questions about how histone modifications come to play a role in the steady state of pluripotency. Another surprise from genetic studies has been the frequency with which the global effects of mutations in chromatin regulators can be largely reversed by a single target gene. These genetic studies help define the arena for future mechanistic studies that might be helpful to harness pluripotency for therapeutic goals.


Antioxidant status and lipid peroxidation in type II diabetes mellitus with and without complications.
1. This study was conducted on 467 cases of non-insulin-dependent diabetes mellitus and 180 healthy controls. Lipid peroxidation products in plasma and erythrocytes were assayed as thiobarbituric acid reactive substances, along with the erythrocyte antioxidant enzymes, namely superoxide dismutase, catalase and glutathione peroxidase. In addition, scavenger vitamins A, C and E and reduced glutathione levels in blood were also measured. 2. Lipid peroxidation was significantly raised within the first 2 years of diagnosis, and superoxide dismutase, catalase, reduced glutathione and vitamins C and E were significantly lowered. 3. These changes were correlated with the duration of the disease and were of a higher magnitude with the development of complications. 4. The results suggest that the antioxidant deficiency and excessive peroxide-mediated damage may appear early on in non-insulin-dependent diabetes mellitus, before the development of secondary complications.


Bisphenol A and other phenols in urine from Danish children and adolescents analyzed by isotope diluted TurboFlow-LC-MS/MS.
Bisphenol A (BPA), triclosan (TCS), benzophenone-3 (BP-3), dichoro- and phenyl phenols are industrial chemicals present in numerous consumer products such as polycarbonate plastics, preservatives in personal care products, sun screens, pesticides and fungicides, respectively, and they are all suspected endocrine disrupters. In this study the urinary excretion of eight phenols in Danish children recruited from the general population were investigated. One 24h urine and two consecutive first morning samples were collected from each of 129 healthy Danish children and adolescents (6-21 years). The concentrations of urinary phenols were analyzed by a new on-line TurboFlow-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Most of the analyzed phenols were detectable in more than 80% of the 24h urine samples and the median concentration of BPA, TCS, BP-3, 2,4-dichorophenol and 2,5-dichorophenol (analyzed as ∑DCP), 2-phenylphenol and 4-phenylphenol were 1.37, 1.45, 1.41, 0.65, 0.36 and 0.53ng/mL, respectively. The ranges of the excreted TCS and BP-3 were wide; from below limit of detection to maximum levels of 955ng/mL and 162ng/mL, respectively, while the other phenols were excreted in a more narrow range with maximum levels below 25ng/mL. Concentrations in first morning urine were in general higher than in 24h urine and comprised 30-47% of the absolute amount excreted during 24h. The youngest children aged 6-10 years had a significantly higher urinary BPA concentration (ng/mL) and also a relatively higher daily BPA excretion (ng/kg bw/24h) than the older children and adolescents. The opposite pattern was observed for TCS, BP-3 and ∑DCP for which urinary levels increased significantly with age. No gender difference or associations to pubertal development were observed. In conclusion, our study showed that Danish children were exposed to multiple phenols simultaneously. Small children were relatively more exposed to BPA than older children, while higher exposures to TCS, BP-3 and ∑DCP were seen among adolescents.


Reflections on self in qualitative research: stories of family.
Reflexivity is a qualitative research strategy that addresses our subjectivity as researchers related to people and events that we encounter in the field. Reflexivity also addresses the subjective nature of the research account as a narrative constructed by us as researchers. Reflexivity enhances the quality of research through its ability to extend our understanding of how our positions and interests as researchers affect all stages of the research process. The reflexive account presented here frames the analysis and interpretation of previously published findings on work and play in families by highlighting aspects of the researcher's reflexivity across the entire research process, including situating the study, gaining access, managing self, living in the field, and telling the story. Its purpose is to demonstrate use of reflexivity in qualitative research as a strategy to consider our subjectivity as researchers and serve as signposts for readers about what is happening throughout the research process.


Care Burden and Coping Strategies among Caregivers of Paediatric HIV/AIDS in Northern Uganda: A Cross-Sectional Mixed-Method Study.
Family caregivers provide the bulk of care to children living with HIV. This places an enormous demand and care burden on the caregivers who often struggle to cope in various ways, some of which may be maladaptive. This may adversely affect their quality of care. Very little literature exists in resource-limited contexts on the burden of care experienced by caregivers on whom children living with HIV/AIDS depend for their long-term care. We assessed care burden and coping strategies among the caregivers of paediatric HIV/AIDS patients in Lira district, northern Uganda. A mixed-method cross-sectional study was conducted among 113 caregivers of paediatric HIV patients attending the ART clinic at a tertiary healthcare facility in Lira district, northern Uganda. A consecutive sampling method was used to select participants for the quantitative study, while 15 respondents were purposively sampled for the qualitative data. Quantitative data were collected using standard interviewer-administered questionnaires, while in-depth interview guides were used to collect qualitative data. Data were entered, cleaned, and analysed using SPSS version 23. Qualitative data were analysed thematically. The majority of the caregivers, 65.5% (74), experienced mild-to-moderate burden. The mean burden scores significantly differed by caregivers' age (P=0.017), marital status (P=0.017), average monthly income (P=0.035), and child's school attendance (P=0.039). Accepting social support, seeking spiritual support, and reframing were the three most commonly used strategies for coping. Marital status and occupation were, respectively, positively and negatively correlated with information-seeking as a coping strategy, while monthly income was positively correlated with psychosocial support as a strategy. Seeking community support was negatively correlated with the duration of the child's care. Our findings show that care burden is a common problem among the caregivers of children living with HIV in the study context.


Identification of Rumen Microbial Genes Involved in Pathways Linked to Appetite, Growth, and Feed Conversion Efficiency in Cattle.
The rumen microbiome is essential for the biological processes involved in the conversion of feed into nutrients that can be utilized by the host animal. In the present research, the influence of the rumen microbiome on feed conversion efficiency, growth rate, and appetite of beef cattle was investigated using metagenomic data. Our aim was to explore the associations between microbial genes and functional pathways, to shed light on the influence of bacterial enzyme expression on host phenotypes. Two groups of cattle were selected on the basis of their high and low feed conversion ratio. Microbial DNA was extracted from rumen samples, and the relative abundances of microbial genes were determined via shotgun metagenomic sequencing. Using partial least squares analyses, we identified sets of 20, 14, 17, and 18 microbial genes whose relative abundances explained 63, 65, 66, and 73% of the variation of feed conversion efficiency, average daily weight gain, residual feed intake, and daily feed intake, respectively. The microbial genes associated with each of these traits were mostly different, but highly correlated traits such as feed conversion ratio and growth rate showed some overlapping genes. Consistent with this result, distinct clusters of a coabundance network were enriched with microbial genes identified to be related with feed conversion ratio and growth rate or daily feed intake and residual feed intake. Microbial genes encoding for proteins related to cell wall biosynthesis, hemicellulose, and cellulose degradation and host-microbiome crosstalk (e.g., aguA, ptb, K01188, and murD) were associated with feed conversion ratio and/or average daily gain. Genes related to vitamin B12 biosynthesis, environmental information processing, and bacterial mobility (e.g., cobD, tolC, and fliN) were associated with residual feed intake and/or daily feed intake. This research highlights the association of the microbiome with feed conversion processes, influencing growth rate and appetite, and it emphasizes the opportunity to use relative abundances of microbial genes in the prediction of these performance traits, with potential implementation in animal breeding programs and dietary interventions.


Association of apelin and AF in patients with implanted loop recorders undergoing catheter ablation.
Previous studies showed an association between apelin and atrial fibrillation (AF). The aim of this study was to analyse the effect of pulmonary vein isolation (PVI) in patients with paroxysmal AF on plasmatic apelin concentrations. Nine consecutive patients (aged from 43 to 69 years, 3 females and 6 males) with documented paroxysmal atrial fibrillation and implanted loop recorders (ILR) for continuous ECG monitoring were included in this study. All the patients underwent a radiofrequency catheter ablation with PVI. The plasmatic concentration of apelin increased after PVI. The average plasmatic concentration of apelin before PVI was 0.299 ng/ml (±0.16), 3 months after PVI 0.462 ng/ml (±0.10) and 9 months after PVI 0.565 ng/ml (±0.146). There was an increase in the concentration of apelin 3 months and 9 months after the PVI by 0.163 ng/ml (p=0.07) and by 0.266 ng/ml (p=0.01), respectively. The concentration of apelin inversely correlated with the AF burden (r=-0.44, p=0.03). Our study showed a significant increase in apelin levels after the reduction of AF burden via PVI and an inverse correlation with AF burden. Apelin might be a promising marker of AF (Tab. 2, Fig. 2, Ref. 28).


The relationship of training load to physical-capacity changes during international tours in high-performance junior tennis players.
Given the travel that punctuates junior tennis development, an understanding of the changes in fitness owing to touring and the association between training loads (TLs) and fitness on return is vital. The authors investigated physical-capacity changes from pretour to posttour, determining if those changes were related to the TL of athletes on tour. Thirty junior athletes completed fitness testing before and after 4-wk tours. Testing included double-leg countermovement jump (CMJ), dominant single-leg and nondominant single-leg CMJ, speed (5, 10, 20 m), modified 5-0-5 agility (left and right), 10 × 20-m repeated-sprint ability (RSA), and multistage fitness tests. Repeated-measures ANOVAs determined physical-capacity change, with effect-size analysis establishing the magnitude of change. To avoid regression toward the mean, a 1/3-split technique was implemented for comparative analysis (high to low TLs). Moderate effects (d=0.50-0.70) for reductions of up to 3.6% in 5-, 10-, and 20-m speeds were observed. However, all remaining changes were only of trivial to small magnitude (d<0.40). Closer analysis of the interaction between TL and physical capacities (1/3-split) revealed that subjects who completed the greatest amount of total and tennis TL returned with a greater decline in speed and aerobic capacities (d>0.80). Furthermore, it was observed that match load dictates on- and off-court TL, with an increase in matches won understandably stunting exposure to off-court TL. Specific training should be prescribed on tour to maintain speed characteristics over a 4-wk international tour. On-tour training schedules should be carefully monitored to maximize specific TL exposure after losses on tour.


Preparation and characterization of superparamagnetic molecularly imprinted polymers for selective adsorption and separation of vanillin in food samples.
Novel water-compatible superparamagnetic molecularly imprinted polymers (M-MIPs) were prepared by coating superparamagnetic Fe3O4 nanoparticles with MIPs in a methanol-water reaction system. The M-MIPs were used for the selective adsorption and separation of vanillin from aqueous solution. The M-MIPs were characterized by X-ray powder diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), vibrating sample magnetometry (VSM), and scanning electron microscopy (SEM). Results indicated that a core-shell structure of M-MIPs was obtained by coating a layer of silica and MIPs on the surface of the Fe3O4 nanoparticles. The obtained M-MIPs possess a loose and porous structure and can be rapidly separated from the solution using a magnet. The adsorption experiments showed that the binding capacity of the M-MIPs was significantly higher than that of the superparamagnetic non-molecularly imprinted polymers (M-NIPs). Meanwhile, the adsorption of M-MIPs reached equilibrium within 100 min, and the apparent maximum adsorption quantity (Qmax) and dissociation constant (Kd) were 64.12 μmol g(-1) and 58.82 μmol L(-1), respectively. The Scatchard analysis showed that homogeneous binding sites were formed on the M-MIP surface. The recoveries of 83.39-95.58% were achieved when M-MIPs were used for the pre-concentration and selective separation of vanillin in spiked food samples. These results provided the possibility for the separation and enrichment of vanillin from complicated food matrices by M-MIPs.


The fate of papers rejected by Australian Family Physician.
Research papers submitted to Australian Family Physician (AFP) are accepted or rejected on the judgment of the research editor with advice from expert reviewers. Rejection can be outright (e.g., when research is "fatally flawed") or, more often, conditional (when authors are invited to respond to criticisms). Sometimes authors fail to resubmit. The fate of both groups of papers is unknown, as are the reasons for failing to resubmit. We sent an explanatory email to all authors who submitted a paper to the AFP research section between 2002 and 2004, with a simple eight question survey (plus 1-2 additional questions for authors of rejected/withdrawn articles. Of 123 requests sent, 50 were returned (41% response rate). These were supplemented by an extra 19 papers identified by literature searching. Authors of accepted papers were more likely to participate than those whose papers were rejected or withdrawn. Most papers (28/47, 60%) submitted to AFP were written specifically for the journal. Those that were published underwent major change from the original submitted. Three out of 11 papers rejected by AFP were published in another journal. Authors who failed to resubmit (or withdrew) their paper usually cited being too busy. The editorial and peer review process was considered valuable by 74% of responders. Most accepted papers (20/37, 54%) underwent one revision; rejected articles were usually rejected outright (9/11, 82%). That authors often lose interest in getting their paper published after preparing it for submission is curious. Most authors consider peer and editorial review to be valuable.


Gender-related differences in cue-elicited cravings in Internet gaming disorder: The effects of deprivation.
Online gaming has become a popular leisure activity, in which males more frequently develop Internet gaming disorder (IGD) compared to females. However, gender-related neurocognitive differences have largely not been systematically investigated in IGD. Cue-elicited-craving tasks were performed before game playing and immediately after deprivation operationalized as a forced break from gaming when the Internet was disconnected. Ninety-nine subjects with IGD (27 males and 22 females) or recreational game use (RGU; 27 males and 23 females) provided functional MRI and subjective data. Analyses investigating effects of group (IGD and RGU) × gender (male and female) at different times (pre-gaming, post-gaming, and post-pre) on cue-elicited craving and brain responses were performed. Correlations between brain responses and subjective measures were calculated. In pre-, post-, and post-pre tests, significant gender-by-group interactions (p < .001, cluster size > 15 voxels) were observed in the left dorsolateral prefrontal cortex (DLPFC). Further analyses of the DLPFC cluster showed that in post-pre comparisons, results were related to less engagement of the DLPFC in IGD, especially in females. In addition, at post-test, significant interactions were observed in the caudate, as females with IGD showed greater activation as compared to those with RGU. The results raise the possibility that women with RGU may show better executive control than men when facing gaming cues, which may provide resiliency against developing IGD; however, once they develop IGD, their gaming may impair their executive control and enhance their cravings for gaming, which may make it more difficult to quit gaming.