Assessment of Coastal Ecosystem Services for Conservation Strategies in South Korea.
Despite the fact that scientific and political consideration for ecosystem services has dramatically increased over the past decade, few studies have focused on marine and coastal ecosystem services for conservation strategies. We used an ecosystem services approach to assess spatial distributions of habitat risks and four ecosystem services (coastal protection, carbon storage, recreation, and aesthetic quality), and explored the tradeoffs among them in coastal areas of South Korea. Additionally, we analyzed how the social and ecological characteristics in coastal areas interact with conservation and development policies by using this approach. We found strong negative associations between the habitat risks and ecosystem services (aquaculture, carbon storage, recreation, and aesthetic quality) across the coastal counties. Our results showed that the intensity of the habitat risks and the provision of ecosystem services were significantly different between reclamation-dominated and conservation-dominated counties, except for coastal vulnerability. A generalized linear model suggested that reclamation projects were dependent on economic efficiency, whereas demographic pressures and habitat conditions influenced the designation of protected areas at a county level. The ecosystem services approach provided guidelines to achieve both sustainable development and environment conservation. By using the approach, we can select the priority areas for developments while we can minimize the degradation of biodiversity and ecosystem services. As cultural ecosystem services are evenly distributed throughout coastal areas of South Korea, decision makers may employ them to improve the conditions of coastal wetlands outside of protected areas.


Specific detection of Bradyrhizobium and Rhizobium strains colonizing rice (Oryza sativa) roots by 16S-23S ribosomal DNA intergenic spacer-targeted PCR.
In addition to forming symbiotic nodules on legumes, rhizobial strains are members of soil or rhizosphere communities or occur as endophytes, e.g., in rice. Two rhizobial strains which have been isolated from root nodules of the aquatic legumes Aeschynomene fluminensis (IRBG271) and Sesbania aculeata (IRBG74) were previously found to promote rice growth. In addition to analyzing their phylogenetic positions, we assessed the suitability of the 16S-23S ribosomal DNA (rDNA) intergenic spacer (IGS) sequences for the differentiation of closely related rhizobial taxa and for the development of PCR protocols allowing the specific detection of strains in the environment. 16S rDNA sequence analysis (sequence identity, 99%) and phylogenetic analysis of IGS sequences showed that strain IRBG271 was related to but distinct from Bradyrhizobium elkanii. Rhizobium sp. (Sesbania) strain IRBG74 was located in the Rhizobium-Agrobacterium cluster as a novel lineage according to phylogenetic 16S rDNA analysis (96.8 to 98.9% sequence identity with Agrobacterium tumefaciens; emended name, Rhizobium radiobacter). Strain IRBG74 harbored four copies of rRNA operons whose IGS sequences varied only slightly (2 to 9 nucleotides). The IGS sequence analyses allowed intraspecies differentiation, especially in the genus Bradyrhizobium, as illustrated here for strains of Bradyrhizobium japonicum, B. elkanii, Bradyrhizobium liaoningense, and Bradyrhizobium sp. (Chamaecytisus) strain BTA-1. It also clearly differentiated fast-growing rhizobial species and strains, albeit with lower statistical significance. Moreover, the high sequence variability allowed the development of highly specific IGS-targeted nested-PCR assays. Strains IRBG74 and IRBG271 were specifically detected in complex DNA mixtures of numerous related bacteria and in the DNA of roots of gnotobiotically cultured or even of soil-grown rice plants after inoculation. Thus, IGS sequence analysis is an attractive technique for both microbial ecology and systematics.


The recovery paradigm - a model of hope and change for alcohol and drug addiction.
Alcohol and drug disorders remain major health and social problems in Australia, contributing enormously to the global burden of disease and the everyday practice of primary care. A recent growth in recovery research and recovery focused policies are starting to have an impact in Australia, with implications for how we attempt to resolve these problems. In this article we discuss recent international findings in recovery research, and explore their implications for primary care. Research indicates that over half of dependent substance users will eventually achieve stable recovery. Key predictors of recovery are active engagement in the community and immersion in peer support groups and activities. Recovery requires a twin track approach: enabling and supporting individual recovery journeys, while creating environmental conditions that enable and support a 'social contagion' of recovery, in which recovery is transmitted through supportive social networks and dedicated recovery groups, such as mutual aid.


Cytoskeleton interruption in human hepatoma HepG2 cells induced by ketamine occurs possibly through suppression of calcium mobilization and mitochondrial function.
Ketamine is an intravenous anesthetic agent often used for inducing and maintaining anesthesia. Cytoskeletons contribute to the regulation of hepatocyte activity of drug biotransformation. In this study, we attempted to evaluate the effects of ketamine on F-actin and microtubular cytoskeletons in human hepatoma HepG2 cells and its possible molecular mechanisms. Exposure of HepG2 cells to ketamine at <or=100 microM, which corresponds to clinically relevant concentrations for 1, 6, and 24 h, did not affect cell viability. Meanwhile, administration of therapeutic concentrations of ketamine obviously interrupted F-actin and microtubular cytoskeletons. In parallel, levels of intracellular calcium concentration- and time-dependently decreased after ketamine administration. Analysis by confocal microscopy further revealed that ketamine suppressed calcium mobilization from an extracellular buffer into HepG2 cells. Exposure to ketamine decreased cellular ATP levels. The mitochondrial membrane potential and complex I NADH dehydrogenase activity were both reduced after ketamine administration. Ketamine did not change the production of actin or microtubulin mRNA in HepG2 cells. Consequently, ketamine-caused cytoskeletal interruption led to suppression of CYP3A4 expression and its metabolizing activity. Therefore, this study shows that therapeutic concentrations of ketamine can disrupt F-actin and microtubular cytoskeletons possibly through suppression of intracellular calcium mobilization and cellular ATP synthesis due to down-regulation of the mitochondrial membrane potential and complex I enzyme activity. Such disruption of the cytoskeleton may lead to reductions in CYP3A4 activity in HepG2 cells.


Effects of single-leg drop-landing exercise from different heights on skeletal adaptations in prepubertal girls: a randomized controlled study.
Few studies have explored osteogenic potential of prepubertal populations. We conducted a 28-week school-based exercise trial of single-leg drop-landing exercise with 42 prepubertal girls (6 to 10 years) randomly assigned to control (C), low-drop (LD) or high-drop (HD) exercise groups. The latter two groups performed single-leg drop-landings (3 sessions/week-1 and 50 landings/session-1) from 14 cm(LD) and 28 cm(HD) using the nondominant leg. Osteogenic responses were assessed using Dual Energy X-ray Absorptiometry (DXA). Single-leg peak ground-reaction impact forces (PGRIF) in a subsample ranged from 2.5 to 4.4 x body-weight (BW). No differences (p > .05) were observed among groups at baseline for age, stature, lean tissue mass (LTM), leisure time physical activity, or average daily calcium intake. After adjusting for covariates of body mass, fat mass and LTM, no differences were found in bone mineral measures or site-specific bone mineral density (BMD) at the hip and lower leg among exercise or control groups. Combining data from both exercise groups failed to produce differences in bone properties when compared with the control group. No changes were apparent for between-leg differences from baseline to posttraining. In contrast to some reports, our findings suggest that strictly controlled unimodal, unidirectional single-leg drop-landing exercises involving low-moderate peak ground-reaction impact forces are not osteogenic in the developing prepubertal female skeleton.


Gonadal and nongonadal mechanisms contribute to the prepubertal hiatus in gonadotropin secretion in the female rhesus monkey (Macaca mulatta).
The present study reexamined the role of the ovary in determining the prepubertal hiatus of gonadotropin secretion in the rhesus monkey. Day- and nighttime blood samples were obtained weekly from neonatally (7-10 days of age) ovariectomized and intact monkeys from birth until 3 yr of age. In the intact monkeys, plasma FSH levels increased during the first month of life, remained elevated until approximately 3 months of age, and then decreased to become undetectable by 7 months of age. Thereafter, plasma FSH remained undetectable until approximately 19 months of age, at which time it again increased to detectable concentrations. In animals ovariectomized as neonates, the developmental pattern in FSH secretion was similar to that in intact animals, but, quantitatively, mean plasma FSH concentrations in the agonadal females were greater than those in the intact control group at all times. Circulating daytime LH concentrations in intact animals were generally below the sensitivity of the assay during the neonatal and prepubertal phases of development, but after 27 months of age, this plasma hormone was measurable on occasion. In neonatally ovariectomized monkeys, daytime LH was elevated during the first month of life, undetectable between 2-20 months of age, and then rose into the adult range by the end of the study. Nocturnal plasma FSH and LH concentrations in agonadal monkeys were generally greater than those during the day at all stages of development. Of particular note was the finding that during the prepubertal hiatus in gonadotropin secretion, when daytime LH levels were mostly immeasurable, nighttime levels of this gonadotropin were consistently elevated. The hypersecretion of gonadotropin during prepubertal development in agonadal animals also occurred when ovariectomy was performed at 61-62 weeks of age. These findings demonstrate that in the female monkey, the open loop activity of the GnRH pulse generator during juvenile development is only partially suppressed, and the ovary contributes significantly to the prepubertal restraint on gonadotropin secretion. We also report the serendipitous finding that a precipitous, albeit transient, decline in circulating gonadotropin concentrations occurred in juvenile monkeys after separation from their mothers and relocation to individual cages. This suppression, which was accompanied by elevated plasma cortisol levels, was apparently not related to any impairment in growth.


Co-Adaptation Processes of Syntactic Complexity in Real-Time Kindergarten Teacher-Student Interactions.
Under the premise that language learning is bidirectional in nature, this study aimed to investigate syntactic coordination within teacher-student interactions by using cross-recurrence quantification analysis (CRQA). Seven teachers' and a group of their students' interactions were repeatedly measured in the course of an intervention in early science education. Results showed changes in the proportion of recurrent points; in case of simple sentences teachers and students became less coordinated over time, whereas in case of complex sentences teachers and students showed increasing coordination. Results also revealed less rigid (more flexible) syntactic coordination, although there were no changes in the relative contribution of teacher and students to this. In the light of the intervention under investigation this is an important result. This means that teachers and students learn to use more complex language and coordinate their language complexity better in order to co-construct science discourse. The application of CRQA provides new insights and contributes to better understanding of the dynamics of syntactic coordination.


Fetal allogeneic umbilical cord cell transplantation improves motor function in spinal cord-injured rats.
The objective of this study was to investigate whether the transplantation of fetal umbilical cord tissue cells as a source of stem cells into the acutely injured spinal cord would produce some regenerations and/or functional recovery in a rat model of spinal cord injury. Five pregnant albino Wistar rats of 12 days gestation were used for obtaining an umbilical cord cell graft. At the second stage of the experiment only Th8-Th9 laminectomy was performed in Group A animals while Group B animals underwent spinal cord hemitransection. The cultured fetal umbilical cord cells coated with Alginate Gel were placed into the lesion cavity immediately after surgery in Group C animals. Group D animals received only Alginate gel sponges into the injured area. All experiment groups were analyzed histologically and immunohistochemically (GFAP, Ki-67, and Pan cadherin) and for motor function after surgery. The umbilical cord cell transplanted animals showed a significant motor recovery compared to non-transplanted animals at 8 and 21 days after spinal cord injury (p=0.008). Significant GFAP and Ki-67 expressions were noted in transplanted animals (p=0.048) suggesting astroglial proliferation. Our findings support the possibility of some functional recovery after umbilical cord cell transplantation following spinal cord injury.


More Unnecessary Imaginary Worlds - Part 3: Cystic Fibrosis and the Institute for Clinical and Economic Review's Draft Evidence Report.
On 20 February 2020, the Institute for Clinical and Economic Review (ICER) released its draft evidence report to establish the value of innovative therapies in the treatment of cystic fibrosis. Following its usual practice, ICER contracted with an outside group to construct a value assessment framework, in this case a microsimulation model, to generate value claims. The primary outcomes for value claims were incremental cost-per-QALY simulations for four target cystic fibrosis populations. The value assessment, in common with the same model applied earlier by ICER in cystic fibrosis, recommended substantial price discounts based on arbitrary threshold cost-per-QALY values. Unfortunately, the entire exercise, as detailed in previous commentaries in INNOVATIONS in Pharmacy is essentially a waste of time. Not only is the reference case model presented by ICER only one of a multiverse of other models, all driven by a selective application of model structure and assumptions, but the fact that the utilities that are applied to hypothetical time spent in different disease stages to modeled QALYs and lifetime cost-per-QALY claims fail to meet fundamental measurement axioms: they are ordinal manifest scores. Applied to target cystic fibrosis target patient groups, the modeled claims are meaningless. From the manufacturer's perspective, in this case Vertex Pharmaceuticals who have developed all the cystic fibrosis therapies 'modeled' by the ICER contractor, their response to ICER claims should be to reject them out of hand; the constructs are imaginary and the outcome claims nonsense.


Different anti-HCV profiles of statins and their potential for combination therapy with interferon.
We recently developed a genome-length hepatitis C virus (HCV) RNA replication system (OR6) with luciferase as a reporter. The OR6 assay system has enabled prompt and precise quantification of HCV RNA replication. Pegylated interferon (IFN) and ribavirin combination therapy is the world standard for chronic hepatitis C, but its effectiveness is limited to about 55% of patients. Newer therapeutic approaches are needed. In the present study, we used the OR6 assay system to evaluate the anti-HCV activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, called statins, and their effects in combination with IFN-alpha. Five types of statins (atorvastatin, fluvastatin, lovastatin, pravastatin, and simvastatin) were examined for their anti-HCV activities. Fluvastatin exhibited the strongest anti-HCV activity (IC50: 0.9 micromol/L), whereas atorvastatin and simvastatin showed moderate inhibitory effects. However, lovastatin, reported recently as an inhibitor of HCV replication, was shown to exhibit the weakest anti-HCV activity. The anti-HCV activities of statins were reversed by the addition of mevalonate or geranylgeraniol. Surprisingly, however, pravastatin exhibited no anti-HCV activity, although it worked as an inhibitor for HMG-CoA reductase. The combination of IFN and the statins (except for pravastatin) exhibited strong inhibitory effects on HCV RNA replication. In combination with IFN, fluvastatin also exhibited a synergistic inhibitory effect. In conclusion, statins, especially fluvastatin, could be potentially useful as new anti-HCV reagents in combination with IFN.


Epidemiological overview of metastatic ovarian carcinoma: long-term experience of TOTSG database.
Malignant ovarian neoplasm is one of the most lethal malignancies among cancers of the female reproductive system. Occasionally, these tumors originate from non-ovarian organs as metastatic lesions since the ovary is a frequent metastatic target of many cancers. However, there limited clinical information on metastatic ovarian carcinoma (MOC) and its hallmarks are unknown. During the period of 1986-2015, 4,284 patients with malignant ovarian neoplasm were identified using the Tokai Ovarian Tumor Study Group (TOTSG) database. Of these, excluding borderline malignant tumor, 3,478 patients with malignant ovarian cancer were extracted. The pathological slides were evaluated under central pathological review. Among them, a total of 143 (4.1%) patients with MOC were identified. The median age of patients with MOC was 54 (29-82) years. The most and second most frequent original tumors were colorectal (43%, N=62) and gastric (29%, N=42) carcinoma, respectively. The rates of carcinoma of the appendix, breast, and pancreas were 8, 6, and 4%, respectively. This is the one of the largest studies clarifying the rates of MOC among malignant ovarian neoplasms. Although the rate is low, we should keep in mind that MOC, particularly from colorectal and gastric cancer should be considered when encountering clinical practice of ovarian cancer.


From parent to child? Transmission of educational attainment within immigrant families: methodological considerations.
One in five U.S. residents under the age of 18 has at least one foreign-born parent. Given the large proportion of immigrants with very low levels of schooling, the strength of the intergenerational transmission of education between immigrant parent and child has important repercussions for the future of social stratification in the United States. We find that the educational transmission process between parent and child is much weaker in immigrant families than in native families and, among immigrants, differs significantly across national origins. We demonstrate how this variation causes a substantial overestimation of the importance of parental education in immigrant families in studies that use aggregate data. We also show that the common practice of "controlling" for family human capital using parental years of schooling is problematic when comparing families from different origin countries and especially when comparing native and immigrant families. We link these findings to analytical and empirical distinctions between group- and individual-level processes in intergenerational transmission.


Tamm-Horsfall protein is a critical renal defense factor protecting against calcium oxalate crystal formation.
The tubular fluid of the mammalian kidney is often supersaturated with mineral salts, but crystallization rarely occurs under normal conditions. The unique ability of the kidney to avoid harmful crystal formation has long been attributed to the inhibitory activity of the urinary macromolecules, although few in vivo studies have been carried out to examine this hypothesis. Here we examined the role of Tamm-Horsfall protein (THP), the principal urinary protein, in urinary defense against renal calcium crystal formation, using a THP knockout model that we recently developed. Wild-type and THP knockout mice were examined for the spontaneous formation of renal calcium crystals using von Kossa staining. The susceptibility of these mice to experimentally induced renal crystal formation was evaluated by administering mice with ethylene glycol, a precursor of oxalate, and vitamin D(3), which increases calcium absorption. Renal calcium crystals were visualized by von Kossa stain, dark field microscopy with polarized light and scanning electron microscopy. Inactivating the THP gene in mouse embryonic stem cells results in spontaneous formation of calcium crystals in adult kidneys. Excessive intake of calcium and oxalate, precursors of the most common type of human renal stones, dramatically increases both the frequency and the severity of renal calcium crystal formation in THP-deficient, but not in wild-type mice. Under high calcium/oxalate conditions, the absence of THP triggers a marked, adaptive induction in renal epithelial cells of osteopontin (OPN), a potent inhibitor of bone mineralization and vascular calcification. Thus, OPN may serve as an inducible inhibitor of calcium crystallization, whereas THP can serve as a constitutive and apparently more effective inhibitor. These results provide the first in vivo evidence that THP is a critical urinary defense factor and suggest that its deficiency could be an important contributing factor in human nephrolithiasis, a condition afflicting tens of millions of people in the world annually.


Synthetic virus-like particles target dendritic cell lipid rafts for rapid endocytosis primarily but not exclusively by macropinocytosis.
DC employ several endocytic routes for processing antigens, driving forward adaptive immunity. Recent advances in synthetic biology have created small (20-30 nm) virus-like particles based on lipopeptides containing a virus-derived coiled coil sequence coupled to synthetic B- and T-cell epitope mimetics. These self-assembling SVLP efficiently induce adaptive immunity without requirement for adjuvant. We hypothesized that the characteristics of DC interaction with SVLP would elaborate on the roles of cell membrane and intracellular compartments in the handling of a virus-like entity known for its efficacy as a vaccine. DC rapidly bind SVLP within min, co-localised with CTB and CD9, but not caveolin-1. In contrast, internalisation is a relatively slow process, delivering SVLP into the cell periphery where they are maintained for a number of hrs in association with microtubules. Although there is early association with clathrin, this is no longer seen after 10 min. Association with EEA-1(+) early endosomes is also early, but proteolytic processing appears slow, the SVLP-vesicles remaining peripheral. Association with transferrin occurs rarely, and only in the periphery, possibly signifying translocation of some SVLP for delivery to B-lymphocytes. Most SVLP co-localise with high molecular weight dextran. Uptake of both is impaired with mature DC, but there remains a residual uptake of SVLP. These results imply that DC use multiple endocytic routes for SVLP uptake, dominated by caveolin-independent, lipid raft-mediated macropinocytosis. With most SVLP-containing vesicles being retained in the periphery, not always interacting with early endosomes, this relates to slow proteolytic degradation and antigen retention by DC. The present characterization allows for a definition of how DC handle virus-like particles showing efficacious immunogenicity, elements valuable for novel vaccine design in the future.


Equating accelerometer estimates among youth: The Rosetta Stone 2.
Different accelerometer cutpoints used by different researchers often yields vastly different estimates of moderate-to-vigorous intensity physical activity (MVPA). This is recognized as cutpoint non-equivalence (CNE), which reduces the ability to accurately compare youth MVPA across studies. The objective of this research is to develop a cutpoint conversion system that standardizes minutes of MVPA for six different sets of published cutpoints. Secondary data analysis. Data from the International Children's Accelerometer Database (ICAD; Spring 2014) consisting of 43,112 Actigraph accelerometer data files from 21 worldwide studies (children 3-18 years, 61.5% female) were used to develop prediction equations for six sets of published cutpoints. Linear and non-linear modeling, using a leave one out cross-validation technique, was employed to develop equations to convert MVPA from one set of cutpoints into another. Bland Altman plots illustrate the agreement between actual MVPA and predicted MVPA values. Across the total sample, mean MVPA ranged from 29.7MVPAmind(-1) (Puyau) to 126.1MVPAmind(-1) (Freedson 3 METs). Across conversion equations, median absolute percent error was 12.6% (range: 1.3 to 30.1) and the proportion of variance explained ranged from 66.7% to 99.8%. Mean difference for the best performing prediction equation (VC from EV) was -0.110mind(-1) (limits of agreement (LOA), -2.623 to 2.402). The mean difference for the worst performing prediction equation (FR3 from PY) was 34.76mind(-1) (LOA, -60.392 to 129.910). For six different sets of published cutpoints, the use of this equating system can assist individuals attempting to synthesize the growing body of literature on Actigraph, accelerometry-derived MVPA.


Role of endogenous incretins in the regulation of postprandial lipoprotein metabolism.
Incretins are known to influence lipid metabolism in the intestine when administered as pharmacologic agents. The aggregate influence of endogenous incretins on chylomicron production and clearance is less clear, particularly in light of opposing effects of co-secreted hormones. Here, we tested the hypothesis that physiological levels of incretins may impact on production or clearances rates of chylomicrons and VLDL. A group of 22 overweight/obese men was studied to determine associations between plasma levels of glucagon-like peptides 1 and 2 (GLP-1 and GLP-2) and glucose-dependent insulinotropic polypeptide (GIP) after a fat-rich meal and the production and clearance rates of apoB48- and apoB100-containing triglyceride-rich lipoproteins. Subjects were stratified by above- and below-median incretin response (area under the curve). Stratification yielded subgroups that differed about two-fold in incretin response. There were neither differences in apoB48 production rates in chylomicrons or VLDL fractions nor in apoB100 or triglyceride kinetics in VLDL between men with above- vs below-median incretin responses. The men with above-median GLP-1 and GLP-2 responses exhibited higher postprandial plasma and chylomicron triglyceride levels, but this could not be related to altered kinetic parameters. No differences were found between incretin response subgroups and particle clearance rates. We found no evidence for a regulatory effect of endogenous incretins on contemporaneous chylomicron or VLDL metabolism following a standardised fat-rich meal. The actions of incretins at pharmacological doses may not be reflected at physiological levels of these hormones.


Minor physical anomalies and their relationship to the aetiology of schizophrenia.
Previous reports have suggested an increased prevalence of minor physical anomalies (MPAs) in schizophrenia. A review of the literature was performed to examine the relationship between MPAs, schizophrenia and other disorders. Previous studies demonstrate a number of methodological shortcomings. Moreover, MPAs are found in several other psychiatric disorders. Proponents of the neurodevelopmental model of schizophrenia have focused on an environmental explanation for the increased prevalence of MPAs. We argue that this may be overly simplistic and propose various ways in which genetic factors may also be involved. Better studies should be performed to examine more precisely the nature of MPAs in schizophrenia and other disorders and also the relationship between MPAs and other developmental abnormalities. At present, it is unclear if MPAs are directly related to the pathogenesis of the disorder or even if they are related to the timing of critical events.


The Impact of Solvent Vapor on the Film Morphology and Crystallization Kinetics of Lead Halide Perovskites during Annealing.
One of the key factors for the remarkable improvements of halide perovskite solar cells over the last few years is the increased control over perovskite crystallinity and its thin film morphology. Among various processing methods, solvent vapor-assisted annealing (SVAA) has proven to be promising in achieving high-quality perovskite films. However, a comprehensive understanding of the perovskite crystallization process during SVAA is still lacking. In this work, we use a home-built setup to precisely control the SVAA conditions to investigate in detail the perovskite crystallization kinetics. By changing the solvent vapor concentration during annealing, the perovskite grain size can be tuned from 200 nm to several micrometers. We monitor the crystallization kinetics during solvent-free annealing and SVAA using in situ grazing incidence wide-angle X-ray scattering, where we find a diminished perovskite growth rate and the formation of low dimensional perovskite at the top of the perovskite layer during SVAA. Scanning electron microscopy images of the final films further suggest that the perovskite growth follows an Ostwald ripening process at higher solvent concentrations. Thus, our results will contribute to achieve a more targeted processing of perovskite films.


Multi-center evaluation of oxidation-reduction potential by the MiOXSYS in males with abnormal semen.
According to the World Health Organization (WHO), oxidative stress (OS) is a significant contributor to male infertility. Seminal OS can be measured by a number of assays, all of which are either costly or time sensitive and/or require large semen volume and complex instrumentation. One less expensive alternative is to quantify the oxidation-reduction potential (ORP) with the MiOXSYS. In this international multi-center study, we assessed whether ORP levels measured by the MiOXSYS could distinguish semen samples that fall within the 2010 WHO normal reference values from those that do not. Semen samples were collected from 2092 patients in 9 countries; ORP was normalized to sperm concentration (mV/106 sperm/ml). Only those samples with a concentration >1 × 106 sperm ml-1 were included. The results showed that 199 samples fell within the WHO normal reference range while the remaining 1893 samples did not meet one or more of the criteria. ORP was negatively correlated with all semen parameters (P < 0.01) except volume. The area under the curve for ORP was 0.765. The ORP cut-off value (1.34 mV/106 sperm/ml) was able to differentiate specimens with abnormal semen parameters with 98.1% sensitivity, 40.6% specificity, 94.7% positive predictive value (PPV) and 66.6% negative predictive value (NPV). When used as an adjunct to traditional semen analysis, ORP levels may help identify altered functional status of spermatozoa caused by OS in cases of idiopathic male infertility and in male partners of couples suffering recurrent pregnancy loss, and thereby directing these men to relevant medical therapies and lifestyle modifications.


Fluorinated dendritic amphiphiles, their stomatosome aggregates and application in enzyme encapsulation.
Enzymes are more selective and efficient than synthetic catalysts but are limited by difficult recycling. This is overcome by immobilisation, namely through encapsulation, with the main drawback of this method being slow diffusion of products and reactants, resulting in effectively lowered enzyme activity. Fluorinated dendritic amphiphiles were reported to self-assemble into regularly perforated bilayer vesicles, so-called "stomatosomes". It was proposed that they could be promising novel reaction vessels due to their increased porosity while retaining larger biomolecules at the same time. Amphiphiles were synthesised and their aggregation was analysed by cryogenic transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS) in buffered conditions necessary for enzyme encapsulation. Urease and albumin were encapsulated using the thin-film hydration method and investigated by confocal and time-gated stimulated emission depletion microscopy (gSTED). Their release was then used to probe the selective retention of cargo by stomatosomes. Free and encapsulated enzyme activity were compared and their capacity to be reused was evaluated using the Berthelot method. Urease was successfully encapsulated, did not leak out at room temperature, and showed better activity in perforated vesicles than in closed vesicles without perforations. Encapsulated enzyme could be reused with retained activity over 8 cycles using centrifugation, while free enzyme had to be filtrated. These results show that stomatosomes may be used in enzyme immobilisation applications and present advantages over closed vesicles or free enzyme.


Systems assessment of transcriptional regulation on central carbon metabolism by Cra and CRP.
Two major transcriptional regulators of carbon metabolism in bacteria are Cra and CRP. CRP is considered to be the main mediator of catabolite repression. Unlike for CRP, in vivo DNA binding information of Cra is scarce. Here we generate and integrate ChIP-exo and RNA-seq data to identify 39 binding sites for Cra and 97 regulon genes that are regulated by Cra in Escherichia coli. An integrated metabolic-regulatory network was formed by including experimentally-derived regulatory information and a genome-scale metabolic network reconstruction. Applying analysis methods of systems biology to this integrated network showed that Cra enables optimal bacterial growth on poor carbon sources by redirecting and repressing glycolysis flux, by activating the glyoxylate shunt pathway, and by activating the respiratory pathway. In these regulatory mechanisms, the overriding regulatory activity of Cra over CRP is fundamental. Thus, elucidation of interacting transcriptional regulation of core carbon metabolism in bacteria by two key transcription factors was possible by combining genome-wide experimental measurement and simulation with a genome-scale metabolic model.


Aiding troubled employees: the prevalence, cost, and characteristics of employee assistance programs in the United States.
Employee assistance programs (EAPs) are job-based programs designed to identify and assist troubled employees. This study determines the prevalence, cost, and characteristics of these programs in the United States by worksite size, industry, and census region. A stratified national probability sample of more than 6400 private, nonagricultural US worksites with 50 or more full-time employees was contacted with a computer-assisted telephone interviewing protocol. More than 3200 worksites responded and were eligible, with a response rate of 90%. Approximately 33% of all private, nonagricultural worksites with 50 or more full-time employees currently offer EAP services to their employees, an 8.9% increase over 1985. These programs are more likely to be found in larger worksites and in the communications/utilities/transportation industries. The most popular model is an external provider, and the median annual cost per eligible employee for internal and external programs was $21.83 and $18.09, respectively. EAPs are becoming a more prevalent point of access to health care for workers with personal problems such as substance abuse, family problems, or emotional distress.


Recruitment order of the abdominal muscles varies with postural task.
Abdominal muscle recruitment strategies in response to a postural perturbation contradict the theory that the deeper abdominal muscles are always recruited in advance of the more superficial muscles. The purpose of this study was to determine whether such contrasting muscle recruitment patterns are due to the postural task or the predictability of a postural task. Participants performed an arm raise task as well as an unpredictable and a predictable balance perturbation task (i.e. support-surface translation) while intramuscular electromyographic (EMG) recordings were obtained from the deep [transversus abdominis (TrA)] and superficial [obliquus externus (OE)] abdominal muscles. The abdominal muscle recruitment order was dependent on the postural task but not on the predictability of a postural perturbation. Whereas arm raises elicited similar EMG onset latencies in TrA and OE, the OE onset latency was 48 ms earlier than the TrA following an unpredictable translation (P = 0.003). The early OE activation persisted when the translation was made predictable to the participant (P = 0.024). These results provide evidence that the abdominal muscle recruitment order varies with the trunk stability requirements specific to each task. Rehabilitation strategies focusing on an early TrA activation to improve postural stability may not be appropriate for all everyday tasks.


Intimate partner violence towards women.
Between 10% and 70% of women may have been physically or sexually assaulted by a partner at some stage, with assault rates against men reported at about one quarter of the rate against women. In at least half of people studied, the problem lasts for 5 years or more. Women reporting intimate partner violence (IPV) are more likely than other women to complain of poor physical or mental health, and of disability. We conducted a systematic review and aimed to answer the following clinical question: What are the effects of interventions initiated by healthcare professionals aimed at female victims of intimate partner violence? We searched: Medline, Embase, The Cochrane Library, and other relevant databases up to September 2009 (Clinical Evidence reviews are updated periodically, please check our website for the most up-to-date version of this review). We found 26 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. In this systematic review we present information relating to the effectiveness and safety of the following interventions: advocacy; career counselling plus critical consciousness awareness; cognitive behavioural counselling; cognitive trauma therapy; counselling; nurse support and guidance; peer support groups; safety planning; and shelters.


Antimonene with two-orders-of-magnitude improved stability for high-performance cancer theranostics.
Although the antimonene (AM) nanomaterial is recently emerging as a new photothermal therapy (PTT) agent, its rapid degradation in physiological medium immensely limits its direct utilization. To this end, we herein engineered AM by the cooperation of dimension optimization, size control, and cell membrane (CM) camouflage. Compared with traditional AM nanosheets, the resulting AM nanoparticles (∼55 nm) cloaked with the CM (denoted as CmNPs) exhibited significantly improved stability and increased photothermal efficacy as well as superior tumor targeting capacity. After intravenous injection, the CmNPs enabled satisfactory photoacoustic/photothermal multimodal imaging at tumor sites. Meanwhile, the PTT together with the newly explored function of photodynamic therapy (PDT) achieved a potent combination therapy with few side effects. The maximized theranostic performance thus strongly recommends CmNPs as a safe and highly reliable modality for anticancer therapy.


The in vivo effect of chelidonine on the stem cell system of planarians.
The presence of adult pluripotent stem cells and the amazing regenerative capabilities make planarian flatworms an extraordinary experimental model to assess in vivo the effects of substances of both natural and synthetic origin on stem cell dynamics. This study focuses on the effects of chelidonine, an alkaloid obtained from Chelidonium majus. The expression levels of molecular markers specific for stem or differentiated cells were compared in chelidonine-treated and control planarians. The use of these markers demonstrates that chelidonine produces in vivo a significant anti-proliferative effect on planarian stem cells in a dose-dependent fashion. In response to chelidonine treatment mitotic abnormalities were also observed and the number of cells able to proceed to anaphase/telophase appeared significantly reduced with respect to the controls. Our results support the possibility that chelidonine acts on cell cycle progression by inhibition of tubulin polymerization. These studies provide a basis for preclinical evaluation in vivo of the effects of chelidonine on physiologically proliferating stem cells.


Light-induced conformational changes of LOV1 (light oxygen voltage-sensing domain 1) and LOV2 relative to the kinase domain and regulation of kinase activity in Chlamydomonas phototropin.
Phototropin (phot), a blue light (BL) receptor in plants, has two photoreceptive domains named LOV1 and LOV2 as well as a Ser/Thr kinase domain (KD) and acts as a BL-regulated protein kinase. A LOV domain harbors a flavin mononucleotide that undergoes a cyclic photoreaction upon BL excitation via a signaling state in which the inhibition of the kinase activity by LOV2 is negated. To understand the molecular mechanism underlying the BL-dependent activation of the kinase, the photochemistry, kinase activity, and molecular structure were studied with the phot of Chlamydomonas reinhardtii. Full-length and LOV2-KD samples of C. reinhardtii phot showed cyclic photoreaction characteristics with the activation of LOV- and BL-dependent kinase. Truncation of LOV1 decreased the photosensitivity of the kinase activation, which was well explained by the fact that the signaling state lasted for a shorter period of time compared with that of the phot. Small angle x-ray scattering revealed monomeric forms of the proteins in solution and detected BL-dependent conformational changes, suggesting an extension of the global molecular shapes of both samples. Constructed molecular model of full-length phot based on the small angle x-ray scattering data proved the arrangement of LOV1, LOV2, and KD for the first time that showed a tandem arrangement both in the dark and under BL irradiation. The models suggest that LOV1 alters its position relative to LOV2-KD under BL irradiation. This finding demonstrates that LOV1 may interact with LOV2 and modify the photosensitivity of the kinase activation through alteration of the duration of the signaling state in LOV2.


Neurology of developmental dyslexia.
Developmental dyslexia was until recently considered to belong solely in the domain of educational psychology. With the advent of better theories on language and reading, and better methods for assessing the structure and function of living human brains and for determining genetic transmission, dyslexia is now poised to become a focal concern of cognitive neuroscience, neurology, and genetic research. Still unresolved are questions relating to how much a reading disability represents a normal variation or a separate pathological entity, and whether the cognitive disorder is primarily cognitive, or secondary to a disorder in early perception. Recent findings from neuroanatomy, neurophysiology, neuropsychology, and genetics research are reviewed. (This review is an updated version of a review first published in Current Opinion In Neurology and Neurosurgery 1992, 5:71-76.)


The structure of the N-terminal domain of riboflavin synthase in complex with riboflavin at 2.6A resolution.
Riboflavin synthase of Escherichia coli is a homotrimer with a molecular mass of 70 kDa. The enzyme catalyzes the dismutation of 6,7-dimethyl-8-(1'-D-ribityl)-lumazine, affording riboflavin and 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione. The N-terminal segment (residues 1-87) and the C-terminal segment (residues 98-187) form beta-barrels with similar fold and a high degree of sequence similarity. A recombinant peptide comprising amino acid residues 1-97 forms a dimer, which binds riboflavin with high affinity. Here, we report the structure of this construct in complex with riboflavin at 2.6A resolution. It is demonstrated that the complex can serve as a model for ligand-binding in the native enzyme. The structure and riboflavin-binding mode is in excellent agreement with structural information obtained from the native enzyme from Escherichia coli and riboflavin synthase from Schizosaccharomyces pombe. The implications for the binding specificity and the regiospecificity of the catalyzed reaction are discussed.


Promoting Health and Well-Being Through Mobile Health Technology (Roadmap 2.0) in Family Caregivers and Patients Undergoing Hematopoietic Stem Cell Transplantation: Protocol for the Development of a Mobile Randomized Controlled Trial.
Cancer patients who undergo allogeneic hematopoietic stem cell transplantation are among the most medically fragile patient populations with extreme demands for caregivers. Indeed, with earlier hospital discharges, the demands placed on caregivers continue to intensify. Moreover, an increased number of allogeneic hematopoietic stem cell transplantations are being performed worldwide, and this expensive procedure has significant economic consequences. Thus, the health and well-being of family caregivers have attracted widespread attention. Mobile health technology has been shown to deliver flexible, and time- and cost-sparing interventions to support family caregivers across the care trajectory. This protocol aims to leverage technology to deliver a novel caregiver-facing mobile health intervention named Roadmap 2.0. We will evaluate the effectiveness of Roadmap 2.0 in family caregivers of patients undergoing hematopoietic stem cell transplantation. The Roadmap 2.0 intervention will consist of a mobile randomized trial comparing a positive psychology intervention arm with a control arm in family caregiver-patient dyads. The primary outcome will be caregiver health-related quality of life, as assessed by the PROMIS Global Health scale at day 120 post-transplant. Secondary outcomes will include other PROMIS caregiver- and patient-reported outcomes, including companionship, self-efficacy for managing symptoms, self-efficacy for managing daily activities, positive affect and well-being, sleep disturbance, depression, and anxiety. Semistructured qualitative interviews will be conducted among participants at the completion of the study. We will also measure objective physiological markers (eg, sleep, activity, heart rate) through wearable wrist sensors and health care utilization data through electronic health records. We plan to enroll 166 family caregiver-patient dyads for the full data analysis. The study has received Institutional Review Board approval as well as Code Review and Information Assurance approval from our health information technology services. Owing to the COVID-19 pandemic, the study has been briefly put on hold. However, recruitment began in August 2020. We have converted all recruitment, enrollment, and onboarding processes to be conducted remotely through video telehealth. Consent will be obtained electronically through the Roadmap 2.0 app. This mobile randomized trial will determine if positive psychology-based activities delivered through mobile health technology can improve caregiver health-related quality of life over a 16-week study period. This study will provide additional data on the effects of wearable wrist sensors on caregiver and patient self-report outcomes. ClinicalTrials.gov NCT04094844; https://www.clinicaltrials.gov/ct2/show/NCT04094844. PRR1-10.2196/19288.


Plasma concentrations of 25-hydroxyvitamin D among Jordanians: Effect of biological and habitual factors on vitamin D status.
Vitamin D is cutaneously synthesized following sun exposure (vitamin D3) as well as it is derived from dietary intake (vitamin D3 and D2). Vitamin D2 and D3 are metabolized in the liver to 25-hydroxyvitamin D (25(OH)D). This metabolite is considered the functional indicator of vitamin D stores in humans. Since Jordan latitude is 31°N, cutaneous synthesis of vitamin D3 should be sufficient all year round. However, many indications reveal that it is not the case. Thus, this study was conducted to determine the 25(OH)D status among Jordanians. Three hundred healthy volunteers were enrolled in a cross sectional study; 201 females and 99 males. 25(OH)D and calcium concentrations were measured by enzyme linked immunosorbent assay and spectroscopy techniques, respectively. All participants filled a study questionnaire that covered age, sex, height, weight, diet, and dress style for females. Females were divided according to their dress style: Western style, Hijab (all body parts are covered except the face and hands), and Niqab (all body parts are covered including face and hands). The average plasma 25(OH)D levels in males and females were 44.5 ± 10.0 nmol/l and 31.1 ± 12.0 nmol/l, respectively. However, when female 25(OH)D levels were categorized according to dress styles, the averages became 40.3, 31.3 and 28.5 nmol/l for the Western style, Hijab and Niqab groups, respectively. These 25(OH)D levels were significantly less than those of males (p < 0.05, 0.001, 0.001, respectively). In addition, the plasma 25(OH)D levels of the Western style group was significantly higher than those of Hijab and Niqab groups (p < 0.001). Furthermore, dairy consumption in males was a positive significant factor in vitamin D status. Even though calcium concentrations were within the reference range, the Hijab and Niqab-dressed females have significantly less plasma calcium levels than males (p < 0.01). Very low plasma 25(OH)D levels in females wearing Hijab or Niqab are highly attributed to low sunlight or UVB exposure. In addition, most of males (76%) and Western style dressed females (90%) have 25(OH)D concentrations below the international recommended values (50 nmol/l), suggesting that although sun exposure should be enough, other factors do play a role in these low concentrations. These findings emphasize the importance of vitamin D supplementation especially among conservatively dressed females, and determining if single nucleotide polymorphisms of the genes involved in vitamin D metabolism do exist among Jordanians.


Safety and effectiveness of apixaban compared with warfarin among clinically-relevant subgroups of venous thromboembolism patients in the United States Medicare population.
The AMPLIFY trial found significantly lower major bleeding (MB) and similar recurrent venous thromboembolism (VTE) risks associated with apixaban vs warfarin among patients with VTE. To compare MB, clinically-relevant non-major (CRNM) bleeding, and recurrent VTE risks among clinically-relevant subgroups of newly diagnosed elderly patients with VTE prescribed apixaban vs warfarin. US Medicare patients prescribed apixaban or warfarin within 30 days post-VTE encounter were identified. Propensity score matching (PSM) was used to control for patient characteristics. Cox models were used to assess MB, CRNM bleeding, and recurrent VTE. Subgroup analyses were conducted for index VTE encounter type, index VTE diagnosis type, index VTE etiology, sex, and frailty. Post-PSM, 11,363 matched pairs of patients prescribed apixaban or warfarin were identified. Apixaban had lower MB (Hazard Ratio [HR]:0.76; 95% CI:0.64-0.91) and similar recurrent VTE risks (HR:1.04; 95% CI:0.75-1.43) vs warfarin. No significant interactions were observed between treatment and index VTE encounter type, index VTE diagnosis type, or sex for risk of MB, CRNM bleeding, or recurrent VTE. Significant interactions: frail patients prescribed apixaban had a 15% lower, while non-frail patients prescribed apixaban had 32% lower CRNM bleeding risk vs those prescribed warfarin. Patients with provoked VTE prescribed apixaban trended toward a higher, while those with unprovoked VTE trended toward a lower risk of recurrent VTE vs patients prescribed warfarin. Apixaban was associated with significantly lower risks of MB and CRNM bleeding, and similar risk of recurrent VTE as compared with warfarin across the overall population and most subgroups.


Pediatric surgery and parental smoking behavior.
Secondhand smoke exposure poses health risks to children, including increased risks for anesthesia. In adult smokers, surgery serves as a teachable moment to motivate quitting. For parents who smoke, having a child undergo surgery may also serve as a teachable moment for smoking behavioral change. This study determined if there is an association between children undergoing a surgical procedure and changes in their parents' smoking behavior. Secondary analyses were performed using logistic regression analysis of 2005 survey data from the National Health Interview Survey. Analyses included 9,289 parent respondents who provided information on both themselves and their children. Of the sampled children, 1,112 (12.6%, 95% CI:11.7, 13.4) lived in a home with at least one person who smoked inside in a usual week. In multivariate analysis of the relationship between parent and child surgical history in the past 12 months and smoking behavior, surgery in either the parent (odds ratio 2.19, 95% CI: 1.55, 3.08) or child (odds ratio 2.61, 95% CI: 1.56, 4.35) was associated with an increased likelihood of a quit attempt by the parent. However, these attempts were more likely to be successful if the parents (odds ratio 2.35, 95% CI: 1.35, 4.07), not their child (odds ratio 0.51, 95% CI: 0.20, 1.28), had surgery within the past 12 months. Parents who smoke were more likely to make a quit attempt within the past 12 months if their children had surgery within this time, but they were not more likely to succeed in maintaining abstinence and thus could benefit from assistance.


Exchange of peripherally inserted central catheters is associated with an increased risk for bloodstream infection.
It is not uncommon that the peripherally inserted central catheter (PICC) needs to be replaced either due to blockage or migration to a peripheral position. In such circumstances, there are two methods of PICC placement: new-site insertion and exchange by using the old PICC as a guide wire. Our objective was to investigate risk of infection associated with the exchange method. In this retrospective study, data on all PICC insertions in the neonatal intensive care unit in 2004 to 2008 were obtained. In the population who needed removal of existing PICC and insertion of a new one, we compared central line-associated bloodstream infection (CLABSI) within 1 week of insertion between the two insertion methods. Of 1148 PICC insertions reviewed, 164 (103 new-site and 61 exchange insertions) were performed after removal of a blocked/malpositioned PICC and therefore comprised the study population. The rate of CLABSI was higher in the exchange method (9.8% versus 1%, P < 0.007). After adjusting for the confounders, the odds for CLABSI within 7 days of PICC insertion was higher with the exchange method (odds ratio 25.2, 95% confidence interval: 2.17 to 292.98; P = 0.01). In infants, insertion of PICCs using the exchange method carries an increased risk of bloodstream infection.


DNA repair synthesis in human fibroblasts requires DNA polymerase delta.
When UV-irradiated cultured diploid human fibroblasts were permeabilized with Brij-58 then separated from soluble material by centrifugation, conservative DNA repair synthesis could be restored by a soluble factor obtained from the supernatant of similarly treated HeLa cells. Extensive purification of this factor yielded a 10.2 S, 220,000-dalton polypeptide with the DNA polymerase and 3'- to 5'-exonuclease activities reported for DNA polymerase delta II (Crute, J. J., Wahl, A. F., and Bambara, R. A. (1986) Biochemistry 25, 26-36). Monoclonal antibody to KB cell DNA polymerase alpha, while binding to HeLa DNA polymerase alpha, did not bind to the HeLa DNA polymerase delta. Moreover, at micromolar concentrations N2-(p-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate (BuPdGTP) and 2-(p-n-butylanilino)-2'-deoxyadenosine 5'-triphosphate (BuAdATP) were potent inhibitors of DNA polymerase alpha, but did not inhibit the DNA polymerase delta. Neither purified DNA polymerase alpha nor beta could promote repair DNA synthesis in the permeabilized cells. Furthermore, under conditions which inhibited purified DNA polymerase alpha by greater than 90%, neither monoclonal antibodies to DNA polymerase alpha, BuPdGTP, nor BuAdATP was able to inhibit significantly the DNA repair synthesis mediated by the DNA polymerase delta. Thus, it appears that a major portion of DNA repair synthesis induced by UV irradiation might be catalyzed by DNA polymerase delta. When xeroderma pigmentosum human diploid fibroblasts were utilized, DNA repair synthesis dependent upon ultraviolet light could be restored by addition of both T4 endonuclease V and DNA polymerase delta, but not by addition of either one alone. This result suggests that cytosol-depleted permeabilized DNA repair-defective human fibroblasts and HeLa DNA polymerase delta might be exploited to provide a functional assay for purifying active DNA repair factors from DNA repair-proficient cells without a preknowledge of their function.


Implant-supported overdenture therapy: a retrospective study.
All patients (n = 46) treated with implant-supported overdentures at the Department of Prosthetic Dentistry, Dental and Medical Health Centre, Halmstad, Sweden, from 1986 to 1993 were studied. The clinical examination was completed in 1994. The material was divided into two subgroups: Group A had been initially treatment planned for an implant-supported overdenture, and Group B had been planned for fixed prostheses but because of loss of implants before loading, treatment with a fixed prosthesis was not possible. The authors present their experience and patient reactions to overdenture therapy in two defined groups of patients. The implant failure rate before loading for Group A (n = 12) was 15% (six implants out of 39), and the rate before loading for Group B (n = 29) was 43.6% (68 implants out of 156). After prosthodontic treatment in Group A, the implant success rate after loading was 87.9%, and the overdenture stability was 84.6%. In group B, 17 implants placed in the maxillae were lost after overdenture therapy, which resulted in an implant success rate of 79.3%. A total of eight overdentures, all of which had been placed in the maxillae, were lost, resulting in an overdenture stability of 73.3%. In this study "change of retentive clips" was the predominant prosthodontic complication related to the overdentures, especially in Group B. Most of these complications (62%) occurred in patients with clinical signs of bruxism. Patient reactions to treatment with an overdenture were positive regarding esthetics for both groups. More negative views were recorded in Group B than in Group A in response to function and retention of the overdenture.


MetaVelvet: an extension of Velvet assembler to de novo metagenome assembly from short sequence reads.
An important step in 'metagenomics' analysis is the assembly of multiple genomes from mixed sequence reads of multiple species in a microbial community. Most conventional pipelines use a single-genome assembler with carefully optimized parameters. A limitation of a single-genome assembler for de novo metagenome assembly is that sequences of highly abundant species are likely misidentified as repeats in a single genome, resulting in a number of small fragmented scaffolds. We extended a single-genome assembler for short reads, known as 'Velvet', to metagenome assembly, which we called 'MetaVelvet', for mixed short reads of multiple species. Our fundamental concept was to first decompose a de Bruijn graph constructed from mixed short reads into individual sub-graphs, and second, to build scaffolds based on each decomposed de Bruijn sub-graph as an isolate species genome. We made use of two features, the coverage (abundance) difference and graph connectivity, for the decomposition of the de Bruijn graph. For simulated datasets, MetaVelvet succeeded in generating significantly higher N50 scores than any single-genome assemblers. MetaVelvet also reconstructed relatively low-coverage genome sequences as scaffolds. On real datasets of human gut microbial read data, MetaVelvet produced longer scaffolds and increased the number of predicted genes.


nm1054: a spontaneous, recessive, hypochromic, microcytic anemia mutation in the mouse.
Hypochromic, microcytic anemias are typically the result of inadequate hemoglobin production because of globin defects or iron deficiency. Here, we describe the phenotypic characteristics and pathogenesis of a new recessive, hypochromic, microcytic anemia mouse mutant, nm1054. Although the mutation nm1054 is pleiotropic, also resulting in sparse hair, male infertility, failure to thrive, and hydrocephaly, the anemia is the focus of this study. Hematologic analysis reveals a moderately severe, congenital, hypochromic, microcytic anemia, with an elevated red cell zinc protoporphyrin, consistent with functional erythroid iron deficiency. However, serum and tissue iron analyses show that nm1054 animals are not systemically iron deficient. From hematopoietic stem cell transplantation and iron uptake studies in nm1054 reticulocytes, we provide evidence that the nm1054 anemia is due to an intrinsic hematopoietic defect resulting in inefficient transferrin-dependent iron uptake by erythroid precursors. Linkage studies demonstrate that nm1054 maps to a genetic locus not previously implicated in microcytic anemia or iron phenotypes.


The dopaminergic innervation of the pigeon telencephalon: distribution of DARPP-32 and co-occurrence with glutamate decarboxylase and tyrosine hydroxylase.
Dopaminergic axons arising from midbrain nuclei innervate the mammalian and avian telencephalon with heterogeneous regional and laminar distributions. In primate, rodent, and avian species, the neuromodulator dopamine is low or almost absent in most primary sensory areas and is most abundant in the striatal parts of the basal ganglia. Furthermore, dopaminergic fibres are present in most limbic and associative structures. Herein, the distribution of DARPP-32, a phosphoprotein related to the dopamine D1-receptor, was investigated in the pigeon telencephalon by immunocytochemical techniques. Furthermore, co-occurrence of DARPP-32-positive perikarya with tyrosine hydroxylase-positive pericellular axonal "baskets" or glutamate decarboxylase-positive neurons, as well as co-occurrence of tyrosine hydroxylase and glutamate decarboxylase were examined. Specificity of the anti-DARPP-32 monoclonal antibody in pigeon brain was determined by immunoblotting. The distribution of DARPP-32 shared important features with the distribution of D1-receptors and dopaminergic fibres in the pigeon telencephalon as described previously. In particular, DARPP-32 was highly abundant in the avian basal ganglia, where a high percentage of neurons were labelled in the "striatal" parts (paleostriatum augmentatum, lobus parolfactorius), while only neuropil staining was observed in the "pallidal" portions (paleostriatum primitivum). In contrast, DARPP-32 was almost absent or present in comparatively lower concentrations in most primary sensory areas. Secondary sensory and tertiary areas of the neostriatum contained numbers of labelled neurons comparable to that of the basal ganglia and intermediate levels of neuropil staining. Approximately up to one-third of DARPP-32-positive neurons received a basket-type innervation from tyrosine hydroxylase-positive fibres in the lateral and caudal neostriatum, but only about half as many did in the medial and frontal neostriatum, and even less so in the hyperstriatum. No case of colocalization of glutamate decarboxylase and DARPP-32 and no co-occurrence of glutamate decarboxylase-positive neurons and tyrosine hydroxylase-basket-like structures could be detected out of more than 2000 glutamate decarboxylase-positive neurons examined, although the high DARPP-32 and high tyrosine hydroxylase staining density hampered this analysis in the basal ganglia. In conclusion, the pigeon dopaminergic system seems to be organized similar to that of mammals. Apparently, in the telencephalon, dopamine has its primary function in higher level sensory, associative and motor processes, since primary areas showed only weak or no anatomical cues of dopaminergic modulation. Dopamine might exert its effects primarily by modulating the physiological properties of non-GABAergic and therefore presumably excitatory units.


Making research data repositories visible: the re3data.org Registry.
Researchers require infrastructures that ensure a maximum of accessibility, stability and reliability to facilitate working with and sharing of research data. Such infrastructures are being increasingly summarized under the term Research Data Repositories (RDR). The project re3data.org-Registry of Research Data Repositories-has begun to index research data repositories in 2012 and offers researchers, funding organizations, libraries and publishers an overview of the heterogeneous research data repository landscape. In July 2013 re3data.org lists 400 research data repositories and counting. 288 of these are described in detail using the re3data.org vocabulary. Information icons help researchers to easily identify an adequate repository for the storage and reuse of their data. This article describes the heterogeneous RDR landscape and presents a typology of institutional, disciplinary, multidisciplinary and project-specific RDR. Further the article outlines the features of re3data.org, and shows how this registry helps to identify appropriate repositories for storage and search of research data.


Challenges and Opportunities for Tribal Waters: Addressing Disparities in Safe Public Drinking Water on the Crow Reservation in Montana, USA.
Disparities in access to safe public drinking water are increasingly being recognized as contributing to health disparities and environmental injustice for vulnerable communities in the United States. As the Co-Directors of the Apsaálooke Water and Wastewater Authority (AWWWA) for the Crow Tribe, with our academic partners, we present here the multiple and complex challenges we have addressed in improving and maintaining tribal water and wastewater infrastructure, including the identification of diverse funding sources for infrastructure construction, the need for many kinds of specialized expertise and long-term stability of project personnel, ratepayer difficulty in paying for services, an ongoing legacy of inadequate infrastructure planning, and lack of water quality research capacity. As a tribal entity, the AWWWA faces additional challenges, including the complex jurisdictional issues affecting all phases of our work, lack of authority to create water districts, and additional legal and regulatory gaps-especially with regards to environmental protection. Despite these obstacles, the AWWWA and Crow Tribe have successfully upgraded much of the local water and wastewater infrastructure. We find that ensuring safe public drinking water for tribal and other disadvantaged U.S. communities will require comprehensive, community-engaged approaches across a broad range of stakeholders to successfully address these complex legal, regulatory, policy, community capacity, and financial challenges.


Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity.
IL-12, a heterodimeric cytokine, consists of two disulfide-linked subunits, p40 and p35. We investigated the role of p40 in ligand binding and signal transduction by expressing this subunit alone in COS cells. Culture media of the transfected COS cells exhibited specific dose-dependent binding to KIT225/K6 cells, a human T cell line that expresses IL-12R. Analysis of the culture media by SDS-PAGE and Western blotting demonstrated the presence of 40-kDa monomers and 80-kDa disulfide-linked homodimers. The two p40 species were purified and identified by N-terminal sequencing and proteolytic peptide mapping. Characterization of the p40 proteins for binding and bioactivity showed that both the p40 monomer and dimer inhibited 125I-labeled IL-12 binding to IL-12R, but the 80-kDa species, having a 50% inhibitory concentration (IC50) of 20 to 70 ng/ml, was at least 20-fold more effective than the monomer. Although neither the monomer nor the dimer stimulated human PHA-blast proliferation, the 80-kDa dimer inhibited IL-12-induced proliferation in a dose-dependent manner with an IC50 of 65 ng/ml. The results suggest that the IL-12 p40 subunit contains the essential epitopes for receptor binding. However, a proper conformation required for high affinity binding is achieved only when p40 is associated with a p35 subunit or another p40 subunit. When p40 is associated with a p35 subunit, the heterodimer acts as an agonist mediating biologic activity. However, when p40 associates with another p40, the homodimer behaves as an antagonist in vitro.


SARS transmission, risk factors, and prevention in Hong Kong.
We analyzed information obtained from 1,192 patients with probable severe acute respiratory syndrome (SARS) reported in Hong Kong. Among them, 26.6% were hospital workers, 16.1% were members of the same household as SARS patients and had probable secondary infections, 14.3% were Amoy Gardens residents, 4.9% were inpatients, and 9.9% were contacts of SARS patients who were not family members. The remaining 347 case-patients (29.1%) had undefined sources of infection. Excluding those <16 years of age, 330 patients with cases from "undefined" sources were used in a 1:2 matched case-control study. Multivariate analysis of this case-control study showed that having visited mainland China, hospitals, or the Amoy Gardens were risk factors (odds ratio [OR] 1.95 to 7.63). In addition, frequent mask use in public venues, frequent hand washing, and disinfecting the living quarters were significant protective factors (OR 0.36 to 0.58). In Hong Kong, therefore, community-acquired infection did not make up most transmissions, and public health measures have contributed substantially to the control of the SARS epidemic.


Improved Attitudes to Psychiatry: A Global Mental Health Peer-to-Peer E-Learning Partnership.
Health links aim to strengthen healthcare systems in low and middle-income countries through mutual exchange of skills, knowledge, and experience. However, student participation remains limited despite growing educational emphasis upon global health. Medical students continue to report negative attitudes to psychiatry in high-income countries, and in Somaliland, the lack of public sector psychiatrists limits medical students' awareness of mental healthcare. The authors describe the design, implementation, and mixed-methods analysis of a peer-to-peer psychiatry e-learning partnership between UK and Somaliland students arising from a global mental health link between the two countries. Medical students at King's College London and Hargeisa and Amoud universities, Somaliland, were grouped into 24 pairs. Participants aimed to complete ten fortnightly meetings to discuss psychiatry topics via the website MedicineAfrica. Students completed initial and final evaluations including Attitudes toward Psychiatry (ATP-30) questions, a stigma questionnaire, and brief evaluations after each meeting. Quantitative findings demonstrated that enjoyment, interest, and academic helpfulness were rated highly by students in Somaliland and moderately by students in the UK. Somaliland students' attitudes to psychiatry were significantly more positive post-participation, whereas UK students' attitudes remained stable. Qualitative findings identified more gains in factual knowledge for Somaliland students, whereas UK students reported more cross-cultural learning. Reasons for non-completion and student-suggested improvements emphasized the need to ensure commitment to the program by participants. This partnership encouraged students to consider global mental health outside the standard medical education environment, through an e-learning format solely utilizing existing resources. This new approach demonstrates potential benefits to students in contrasting locations of brief, focused online peer-to-peer education partnerships, expanding the scope of health links to the medical professionals of the future.


Fully automated image-based estimation of postural point-features in children with cerebral palsy using deep learning.
The aim of this study was to provide automated identification of postural point-features required to estimate the location and orientation of the head, multi-segmented trunk and arms from videos of the clinical test 'Segmental Assessment of Trunk Control' (SATCo). Three expert operators manually annotated 13 point-features in every fourth image of 177 short (5-10 s) videos (25 Hz) of 12 children with cerebral palsy (aged: 4.52 ± 2.4 years), participating in SATCo testing. Linear interpolation for the remaining images resulted in 30 825 annotated images. Convolutional neural networks were trained with cross-validation, giving held-out test results for all children. The point-features were estimated with error 4.4 ± 3.8 pixels at approximately 100 images per second. Truncal segment angles (head, neck and six thoraco-lumbar-pelvic segments) were estimated with error 6.4 ± 2.8°, allowing accurate classification (F 1 > 80%) of deviation from a reference posture at thresholds up to 3°, 3° and 2°, respectively. Contact between arm point-features (elbow and wrist) and supporting surface was classified at F 1 = 80.5%. This study demonstrates, for the first time, technical feasibility to automate the identification of (i) a sitting segmental posture including individual trunk segments, (ii) changes away from that posture, and (iii) support from the upper limb, required for the clinical SATCo.


Effects of OTU clustering and PCR artifacts on microbial diversity estimates.
Next-generation sequencing has increased the coverage of microbial diversity surveys by orders of magnitude, but differentiating artifacts from rare environmental sequences remains a challenge. Clustering 16S rRNA sequences into operational taxonomic units (OTUs) organizes sequence data into groups of 97 % identity, helping to reduce data volumes and avoid analyzing sequencing artifacts by grouping them with real sequences. Here, we analyze sequence abundance distributions across environmental samples and show that 16S rRNA sequences of >99 % identity can represent functionally distinct microorganisms, rendering OTU clustering problematic when the goal is an accurate analysis of organism distribution. Strict postsequencing quality control (QC) filters eliminated the most prevalent artifacts without clustering. Further experiments proved that DNA polymerase errors in polymerase chain reaction (PCR) generate a significant number of substitution errors, most of which pass QC filters. Based on our findings, we recommend minimizing the number of PCR cycles in DNA library preparation and applying strict postsequencing QC filters to reduce the most prevalent artifacts while maintaining a high level of accuracy in diversity estimates. We further recommend correlating rare and abundant sequences across environmental samples, rather than clustering into OTUs, to identify remaining sequence artifacts without losing the resolution afforded by high-throughput sequencing.


Modeling disease in human ESCs using an efficient BAC-based homologous recombination system.
Although mouse models have been valuable for studying human disease, the cellular and physiological differences between mouse and human have made it increasingly important to develop more relevant human disease models for mechanistic studies and drug discovery. Human embryonic stem cells (hESCs), which can undergo unlimited self-renewal and retain the potential to differentiate into all cell types, present a possible solution. To improve the efficiency of genetic manipulation of hESCs, we have developed bacterial artificial chromosome (BAC) based approach that enables high efficiency homologous recombination. By sequentially disrupting both alleles of ATM or p53 with BAC targeting vectors, we have established ATM(-/-) and p53(-/-) hESCs as models for two major human genetic instability syndromes and used the generated cells to reveal the importance of p53 in maintaining genome stability of hESCs. Our findings suggest that it will be feasible to develop genetically modified hESCs as relevant human disease models.


Role of LRP1 in the pathogenesis of Alzheimer's disease: evidence from clinical and preclinical studies.
Among the LDL receptor (LDLR) family members, the roles of LDLR-related protein (LRP)1 in the pathogenesis of Alzheimer's disease (AD), especially late-onset AD, have been the most studied by genetic, neuropathological, and biomarker analyses (clinical studies) or cellular and animal model systems (preclinical studies) over the last 25 years. Although there are some conflicting reports, accumulating evidence from preclinical studies indicates that LRP1 not only regulates the metabolism of amyloid-β peptides (Aβs) in the brain and periphery, but also maintains brain homeostasis, impairment of which likely contributes to AD development in Aβ-independent manners. Several preclinical studies have also demonstrated an involvement of LRP1 in regulating the pathogenic role of apoE, whose gene is the strongest genetic risk factor for AD. Nonetheless, evidence from clinical studies is not sufficient to conclude how LRP1 contributes to AD development. Thus, despite very promising results from preclinical studies, the role of LRP1 in AD pathogenesis remains to be further clarified. In this review, we discuss the potential mechanisms underlying how LRP1 affects AD pathogenesis through Aβ-dependent and -independent pathways by reviewing both clinical and preclinical studies. We also discuss potential therapeutic strategies for AD by targeting LRP1.


Exclusive selectivity of multidentate ligands independent on the oxidation state of cobalt: influence of steric hindrance on dioxygen binding and phenoxazinone synthase activity.
The present report describes the syntheses, characterizations, crystal structures and study of the phenoxazinone synthase activity of two peroxo-bridged dicobalt(III) complexes, [Co2(L(1))2(μ-O2)](ClO4)4·2CH3CN (1) and [Co2(L(2))2(μ-O2)](ClO4)4 (2), and three mononuclear cobalt(II) complexes, [Co(L(3))(CH3CN)](ClO4)2 (3), [Co(L(4))(H2O)](ClO4)2 (4) and [Co(L(5))(H2O)](ClO4)2 (5), derived from the pentadentate ligands L(1)-L(5), which are the 1 : 2 condensation products of triamines and 2-acetylpyridine or 2-pyridinecarboxaldehyde (6-methyl-2-pyridinecarboxaldehyde). X-ray crystallography reveals exclusive selectivity of the acyclic Schiff dibasic form of the ligands over the heterocyclic analogues, and this selectivity is found to be insensitive to the oxidation state of cobalt. Other first row transition metals have been characterized in either form of the ligands in their complexes but it is specific for cobalt established in the present study. The pronounced effect of the methyl substitutions is observed from their crystal structures; substitution at imine-C does not have any significant influence on the peroxo-bridging but substitution at sixth position of pyridyl ring prevents the formation of peroxo-bridging, and both the steric and electronic factors play vital roles on such chemical diversity. All the complexes show the phenoxazinone synthase mimicking activity and the comparative catalytic activity has been explored. Although electrochemical behaviors of all the complexes are very similar, their relative catalytic activity mimicking the function of phenoxazinone synthase arises from the electronic and steric factors of the methyl substitution.


Improving patient outcomes in fibrous dysplasia/McCune-Albright syndrome: an international multidisciplinary workshop to inform an international partnership.
To develop consensus on improving the management of patients, we convened an international workshop involving patients, clinicians, and researchers. Key findings included the diagnostic delay and variability in subsequent management with agreement to develop an international natural history study. We now invite other stakeholders to join the partnership. The aim of this study was develop a consensus on how to improve the management of patients with fibrous dysplasia and prioritize areas for research METHODS: An international workshop was held over 3 days involving patients, clinicians, and researchers. Each day had a combination of formal presentations and facilitated discussions that focused on clinical pathways and research. The patient workshop day highlighted the variability of patients' experience in getting a diagnosis, the knowledge of general clinical staff, and understanding long-term outcomes. The research workshop prioritized collaborations that improved understanding of the contemporary natural history of fibrous dysplasia/McCune-Albright syndrome (FD/MAS). The clinical workshop outlined the key issues around diagnostics, assessment of severity, treatment and monitoring of patients. In spite of advances in understanding the genetic and molecular underpinnings of fibrous dysplasia/McCune-Albright syndrome, clinical management remains a challenge. From the workshop, a consensus was reached to create an international, multi-stakeholder partnership to advance research and clinical care in FD/MAS. We invite other stakeholders to join the partnership.


Hydrogen peroxide in exhaled air is increased in stable asthmatic children.
Exhaled air condensate provides a noninvasive means of obtaining samples from the lower respiratory tract. Hydrogen peroxide (H2O2) in exhaled air has been proposed as a marker of airway inflammation. We hypothesized that in stable asthmatic children the H2O2 concentration in exhaled air condensate may be elevated as a result of airway inflammation. In a cross-sectional study, 66 allergic asthmatic children (of whom, 41 were treated with inhaled steroids) and 21 healthy controls exhaled through a cold trap. The resulting condensate was examined fluorimetrically for the presence of H2O2. All subjects were clinically stable, nonsmokers, without infection. The median H2O2 level in the exhaled air condensate of the asthmatic patients was significantly higher than in healthy controls (0.60 and 0.15 micromol, respectively; p<0.05), largely because of high values in the stable asthmatic children who did not use anti-inflammatory treatment (0.8 micromol; p<0.01 compared to controls). We conclude that hydrogen peroxide is elevated in exhaled air condensate of children with stable asthma, and may reflect airway inflammation.


A decision framework for estimating the cost of marine plastic pollution interventions.
Marine plastic pollution has emerged as one of the most pressing environmental challenges of our time. Although there has been a surge in global investment for implementing interventions to mitigate plastic pollution, there has been little attention given to the cost of these interventions. We developed a decision support framework to identify the economic, social, and ecological costs and benefits of plastic pollution interventions for different sectors and stakeholders. We calculated net cost as a function of six cost and benefit categories with the following equation: cost of implementing an intervention (direct, indirect, and nonmonetary costs) minus recovered costs and benefits (monetary and nonmonetary) produced by the interventions. We applied our framework to two quantitative case studies (a solid waste management plan and a trash interceptor) and four comparative case studies, evaluating the costs of beach cleanups and waste-to-energy plants in various contexts, to identify factors that influence the costs of plastic pollution interventions. The socioeconomic context of implementation, the spatial scale of implementation, and the time scale of evaluation all influence costs and the distribution of costs across stakeholders. Our framework provides an approach to estimate and compare the costs of a range of interventions across sociopolitical and economic contexts.


Approach to the Patient with Primary Aldosteronism: Utility and Limitations of Adrenal Vein Sampling.
Several studies over the past 3 decades document a higher prevalence of primary aldosteronism (PA) among hypertensive patients than generally presumed. PA exists as a spectrum from mild to severe aldosterone excess. Although a variety of PA subtypes exist, the 2 most common are aldosterone-producing adenomas (APAs) and bilateral hyperaldosteronism (BHA). The distinction is important, because APA-and other subtypes, with aldosterone production mostly from 1 adrenal-can be cured surgically, and BHA should be treated medically with mineralocorticoid-receptor antagonists (MRAs). The major shortcomings in the tailored management of patients with possible PA are the low rates of screening for case identification and the expensive and technically challenging imaging and interventional procedures required to distinguish APA from BHA, especially adrenal vein sampling (AVS). When AVS identifies an APA and allows the patient to be cured surgically, the procedure is of great value. In contrast, the patient with BHA is treated with MRA whether AVS is performed or not. Consequently, it is prudent to gauge how likely it is to benefit from imaging and AVS in each case prior to embarking on these studies. The explosion of information about PA in the past decade, including predictors of APA and of surgical benefit, are useful in limiting the evaluation for some patients with a positive PA screening test. This article will review our suggestions for approaching these patients in a pragmatic style, recognizing the limitations to even the best resources and facilities.


Cellular reorganization in the human retina during normal aging.
To characterize the nature and extent of neuronal reorganization in the human retina during normal aging. Retinas of young (18-34 years old) and aged (68-77 years old) human donors were examined. Immunocytochemical methods and antibodies directed against Go-alpha, protein kinase C, parvalbumin, calbindin, calretinin, and choline acetyltransferase were used to stain different retinal cell types. Confocal images of retinal sections from the optic disc to the peripheral edge were taken at three eccentricities, and the density and length of cellular processes were quantified with neuroanatomical analysis software. Dendritic fibers of rod and On-cone bipolar cells were found to extend well beyond the normal boundary of the outer plexiform layer (OPL) into the outer nuclear layer (ONL) in aged retinas. Length and density of these elongated fibers were significantly greater in aged than in young retinas. This phenomenon demonstrated a clear spatial gradient that was most prevalent in the periphery and was infrequent in the central region of the retina. Horizontal cells, which normally make triad synaptic connections with photoreceptors and bipolar cells, also had dendrites that extended into the ONL in aged retinas, and these were spatially juxtaposed with the elongated dendrites of bipolar cells. Rod and On-cone bipolar cells, as well as horizontal cells of the human retina, undergo extensive dendritic reorganization during normal aging. Although literature on aging has tended to emphasize degenerative and regressive changes, the present findings provide evidence for a remarkable degree of cellular plasticity in the aged human retina.


Oxytocin may mediate the benefits of positive social interaction and emotions.
During breastfeeding or suckling, maternal oxytocin levels are raised by somatosensory stimulation. Oxytocin may, however, also be released by nonnoxious stimuli such as touch, warm temperature etc. in plasma and in cerebrospinal fluid. Consequently, oxytocin may be involved in physiological and behavioral effects induced by social interaction in a more general context. In both male and female rats oxytocin exerts potent physiological antistress effects. If daily oxytocin injections are repeated over a 5-day period, blood pressure is decreased by 10-20 mmHg, the withdrawal latency to heat stimuli is prolonged, cortisol levels are decreased and insulin and cholecystokinin levels are increased. These effects last from 1 to several weeks after the last injection. After repeated oxytocin treatment weight gain may be promoted and the healing rate of wounds increased. Most behavioral and physiological effects induced by oxytocin can be blocked by oxytocin antagonists. In contrast, the antistress effects can not, suggesting that unidentified oxytocin receptors may exist. The prolonged latency in the tail-flick test can be temporarily reversed by administration of naloxone, suggesting that endogenous opioid activity has been increased by the oxytocin injections. In contrast, the long-term lowering of blood pressure and of cortisol levels as well as the sedative effects of oxytocin have been found to be related to an increased activity of central alpha 2-adrenoceptors. Positive social interactions have been related to health-promoting effects. Oxytocin released in response to social stimuli may be part of a neuroendocrine substrate which underlies the benefits of positive social experiences. Such processes may in addition explain the health-promoting effects of certain alternative therapies. Because of the special properties of oxytocin, including the fact that it can become conditioned to psychological state or imagery, oxytocin may also mediate the benefits attributed to therapies such as hypnosis or meditation.


A functional role for endogenous atrial natriuretic peptide in a canine model of early left ventricular dysfunction.
Asymptomatic or early left ventricular dysfunction in humans is characterized by increases in circulating atrial natriuretic peptide (ANP) without activation of the renin-angiotensin-aldosterone system (RAAS). We previously reported a canine model of early left ventricular dysfunction (ELVD) produced by rapid ventricular pacing and characterized by an identical neurohumoral profile and maintenance of the natriuretic response to volume expansion (VE). To test the hypothesis that elevated endogenous ANP suppresses the RAAS and maintains sodium excretion in ELVD, we assessed the effects of antagonism of ANP on cardiorenal and neurohumoral function in ELVD. Chronic ANP suppression was produced by bilateral atrial appendectomies before the production of ELVD by rapid ventricular pacing (ELVD-APPX, n = 5). This group was compared with a separate group with ELVD and intact atrial appendages (ELVD-INTACT, n = 8). ELVD-APPX was characterized by lower circulating ANP (50 +/- 11 vs. 158 +/- 37 pg/ml, P < 0.05), activation of plasma renin activity (PRA) (9.4 +/- 2.4 vs. 0.6 +/- 0.4 ng/ml per h, P < 0.05) and aldosterone (36.4 +/- 12.5 vs. 2.5 +/- 0.0 ng/dl, P < 0.05) when compared to ELVD-INTACT. In comparison to the ELVD-INTACT group, sodium excretion was decreased before and during VE in the ELVD-APPX group. Acute ANP antagonism was produced by administration of the particulate guanylate cyclase coupled natriuretic peptide receptor antagonist, HS-142-1, to seven conscious dogs with ELVD and intact atrial appendages (ELVD-INTACT). HS-142-1 decreased plasma concentrations and renal generation of the ANP second messenger, cGMP, and was associated with activation of PRA and sodium retention with enhanced tubular sodium reabsorption. These data support a significant role for elevated endogenous ANP in the maintenance of sodium excretion and regulation of the RAAS in experimental ELVD.


Mechanical Thrombectomy with the Embolus Retriever with Interlinked Cages in Acute Ischemic Stroke: ERIC, the New Boy in the Class.
The Embolus Retriever with Interlinked Cages (ERIC) device is a novel stent retriever for mechanical thrombectomy. It consists of interlinked cages and could improve procedural benchmarks and clinical outcome compared with classic stent retrievers. This study compares the rates of recanalization, favorable clinical outcome, procedural adverse events, and benchmarks between the ERIC device and classic stent retrievers. From 545 patients treated with thrombectomy between 2012 and 2015, 316 patients were included. The mean age was 69 ±13 years, the mean baseline NIHSS score was 17 ± 5, and 174 (55%) were men. The ERIC was used as the primary thrombectomy device in 59 (19%) patients. In a propensity score matched analysis including the NIHSS score, clot location, delay to groin puncture, neurointerventionalist, and anesthetic management, 57 matched pairs were identified. Patients treated with the ERIC device compared with classic stent retrievers showed equal rates of recanalization (86% versus 81%, P = .61), equal favorable 3-month clinical outcome (mRS 0-2: 46% versus 40%, P = .71), and procedural adverse events (28% versus 30%, P = 1.00). However, in patients treated with the ERIC device, thrombectomy procedures were less time-consuming (67 versus 98 minutes, P = .009) and a rescue device was needed less often (18% versus 39%, P = .02) compared with classic stent retrievers. Mechanical thrombectomy with the ERIC device is effective and safe. Rates of favorable procedural and clinical outcomes are at least as good as those with classic stent retrievers. Of note, the ERIC device might be time-saving and decrease the need for rescue devices. These promising results call for replication in larger prospective clinical trials.


Regulators of autophagosome formation in Drosophila muscles.
Given the diversity of autophagy targets and regulation, it is important to characterize autophagy in various cell types and conditions. We used a primary myocyte cell culture system to assay the role of putative autophagy regulators in the specific context of skeletal muscle. By treating the cultures with rapamycin (Rap) and chloroquine (CQ) we induced an autophagic response, fully suppressible by knockdown of core ATG genes. We screened D. melanogaster orthologs of a previously reported mammalian autophagy protein-protein interaction network, identifying several proteins required for autophagosome formation in muscle cells, including orthologs of the Rab regulators RabGap1 and Rab3Gap1. The screen also highlighted the critical roles of the proteasome and glycogen metabolism in regulating autophagy. Specifically, sustained proteasome inhibition inhibited autophagosome formation both in primary culture and larval skeletal muscle, even though autophagy normally acts to suppress ubiquitin aggregate formation in these tissues. In addition, analyses of glycogen metabolic genes in both primary cultured and larval muscles indicated that glycogen storage enhances the autophagic response to starvation, an important insight given the link between glycogen storage disorders, autophagy, and muscle function.


Myocardial technetium-99m-teboroxime activity in acute coronary artery occlusion and reperfusion: relation to myocardial blood flow and viability.
The purpose of this study was to test the hypothesis that 99mTc-teboroxime retention within the heart depends at least in part on the presence of viable myocytes. We used a porcine model of acute myocardial infarction with reperfusion and compared the myocardial uptake of labeled microspheres at 1 hr of reperfusion with that of 99mTc-teboroxime. Eleven domestic swine had measurements of hemodynamics and regional myocardial blood flow (microspheres) at baseline, at 10 and 50 min of left anterior descending (LAD) coronary artery occlusion and at 10 and 60 min of LAD reperfusion. Technetium-99m-teboroxime was injected intravenously at 60 min of reperfusion and the animal was killed 5-7 min later. The heart was then perfused with triphenyl tetrazolium chloride to identify infarcted and jeopardized myocardium in the occlusion zone and with Evans blue dye to mark normally perfused myocardium. After imaging, tissue sections were digested and colored microspheres were extracted and counted to determine myocardial blood flow. After coronary occlusion, infarct zone (MIZ) to normal zone (NZ) blood flow ratios declined from 0.95 +/- 0.27 (pre-occlusion) to 0.18 +/- 0.15 at 10 min and 0.25 +/- 0.35 at 50 min of occlusion (both p < 0.05). The MIZ:NZ count ratio at 60 min of reperfusion was less than the MIZ:NZ blood flow ratio in every animal and over the entire range of flow ratios (0.55-3.64). Technetium-99m-teboroxime requires viable myocytes for retention within the heart and is not exclusively a tracer of myocardial blood flow when imaged 5-7 min after injection. Additional in vivo imaging studies are required to determine the extent to which reduced retention of the tracer by reperfused but nonviable myocardium influences the appearance of clinical scans.