Report of two patients and further characterization of interstitial 9p13 deletion--a rare but recurrent microdeletion syndrome?
To date, an interstitial deletion of 9p13 has been described only two times in the medical literature. These reports were based on routine chromosomal analysis. We report on two additional patients with an interstitial deletion of 9p13 further defined on array CGH who share clinical features with the other two patients previously described. Our first patient is a 16-year-old girl with a 5.9 Mb deletion at 9p13.3-9p13.1, initially detected on routine karyotype analysis and further characterized on array CGH. Our second patient is a 7½-year-old boy with a 4.8 Mb deletion also at 9p13.3-9p13.1. Patients with 9p13 deletion appear to have mild to moderate developmental delay, social and interactive personality, behavior issues such as attention deficit-hyperactivity disorder, short stature, prominent antihelices, hypoplastic nails, and precocious/early puberty. Our 16-year-old patient is the oldest patient described thus far. This report further characterizes this condition and helps to delineate the long-term prognosis in these patients.


Coproduction of acetaldehyde and hydrogen during glucose fermentation by Escherichia coli.
Escherichia coli K-12 strain MG1655 was engineered to coproduce acetaldehyde and hydrogen during glucose fermentation by the use of exogenous acetyl-coenzyme A (acetyl-CoA) reductase (for the conversion of acetyl-CoA to acetaldehyde) and the native formate hydrogen lyase. A putative acetaldehyde dehydrogenase/acetyl-CoA reductase from Salmonella enterica (SeEutE) was cloned, produced at high levels, and purified by nickel affinity chromatography. In vitro assays showed that this enzyme had both acetaldehyde dehydrogenase activity (68.07 ± 1.63 μmol min(-1) mg(-1)) and the desired acetyl-CoA reductase activity (49.23 ± 2.88 μmol min(-1) mg(-1)). The eutE gene was engineered into an E. coli mutant lacking native glucose fermentation pathways (ΔadhE, ΔackA-pta, ΔldhA, and ΔfrdC). The engineered strain (ZH88) produced 4.91 ± 0.29 mM acetaldehyde while consuming 11.05 mM glucose but also produced 6.44 ± 0.26 mM ethanol. Studies showed that ethanol was produced by an unknown alcohol dehydrogenase(s) that converted the acetaldehyde produced by SeEutE to ethanol. Allyl alcohol was used to select for mutants with reduced alcohol dehydrogenase activity. Three allyl alcohol-resistant mutants were isolated; all produced more acetaldehyde and less ethanol than ZH88. It was also found that modifying the growth medium by adding 1 g of yeast extract/liter and lowering the pH to 6.0 further increased the coproduction of acetaldehyde and hydrogen. Under optimal conditions, strain ZH136 converted glucose to acetaldehyde and hydrogen in a 1:1 ratio with a specific acetaldehyde production rate of 0.68 ± 0.20 g h(-1) g(-1) dry cell weight and at 86% of the maximum theoretical yield. This specific production rate is the highest reported thus far and is promising for industrial application. The possibility of a more efficient "no-distill" ethanol fermentation procedure based on the coproduction of acetaldehyde and hydrogen is discussed.


Multiple sources of slow activity fluctuations in a bacterial chemosensory network.
Cellular networks are intrinsically subject to stochastic fluctuations, but analysis of the resulting noise remained largely limited to gene expression. The pathway controlling chemotaxis of Escherichia coli provides one example where posttranslational signaling noise has been deduced from cellular behavior. This noise was proposed to result from stochasticity in chemoreceptor methylation, and it is believed to enhance environment exploration by bacteria. Here we combined single-cell FRET measurements with analysis based on the fluctuation-dissipation theorem (FDT) to characterize origins of activity fluctuations within the chemotaxis pathway. We observed surprisingly large methylation-independent thermal fluctuations of receptor activity, which contribute to noise comparably to the energy-consuming methylation dynamics. Interactions between clustered receptors involved in amplification of chemotactic signals are also necessary to produce the observed large activity fluctuations. Our work thus shows that the high response sensitivity of this cellular pathway also increases its susceptibility to noise, from thermal and out-of-equilibrium processes.


Lyophilization Based Isolation of Exosomes.
Exosomes are nanoscale extracellular vesicles which regulate intercellular communication. They have great potential for application in nanomedicine. However, techniques for their isolation are limited by requirements for advanced instruments and costly reagents. In this study, we developed a lyophilization-based method for isolating exosomes from cultured cells. The isolated exosomes were characterized for protein content using Bradford assay, and for size distribution and shape using scanning electron microscopy (SEM) and nanoparticles tracking analysis (NTA). In addition, CD63, CD9, CD81, HSP70 and TSG101 were evaluated as essential exosomal surface markers using Western blot. Drug loading and release studies were performed to confirm their drug delivery properties using an in vitro model. Exosomes were also loaded with commercial dyes (Cy5, Eosin) for the evaluation of their drug delivery properties. All these characterizations confirmed successful exosome isolation with measurements of less than 150 nm, having a typical shape, and by expressing the known exosome surface protein markers. Finally, tyrosine kinase inhibitors (dasatinib and ponatinib) were loaded on the exosomes to evaluate their anticancer effects on leukemia cells (K562 and engineered Ba/F3-BCR-ABL) using MTT and Annexin-PI assays. The expression of MUC1 protein on the exosomes isolated from MCF-7 cells also indicated that their potential diagnostic properties were intact. In conclusion, we developed a new method for exosome isolation from cultured cells. These exosomes met all the essential requirements in terms of characterization, drug loading and release ability, and inhibition of proliferation and apoptosis induction in Ph+ leukemia cells. Based on these results, we are confident in presenting the lyophilization-based exosome isolation method as an alternative to traditional techniques for exosome isolation from cultured cells.


HIV-1 is budded from CD4+ T lymphocytes independently of exosomes.
The convergence of HIV-1 budding and exosome biogenesis at late endosomal compartments called multivesicular bodies has fueled the debate on whether HIV-1 is budded from its target cells and transmitted in the form of exosomes. The point of contention appears to primarily derive from the types of target cells in question and lack of a well-defined protocol to separate exosomes from HIV-1. In this study, we adapted and established a simplified protocol to define the relationship between HIV-1 production and exosome biogenesis. Importantly, we took advantage of the newly established protocol to unequivocally show that HIV-1 was produced from CD4+ T lymphocytes Jurkat cells independently of exosomes. Thus, this study not only presents a simplified way to obtain highly purified HIV-1 virions for identification of host proteins packaged into virions, but also provides a technical platform that can be employed to define the relationship between exosome biogenesis and budding of HIV-1 or other viruses and its contributions to viral pathogenesis.


[Language use in medicine].
Language is a main constituent of communication. To understand its signs it is necessary for doctors to use different codes depending on whether they communicate with each other, with their patients, or outsiders. Such codes are every day language, technical language, scientific language and language to the knowledge of different groups of non-specialists.


Effects of tail docking on behavior of confined feedlot cattle.
Tail tip injuries occur in some feedlot cattle housed in slatted-floor facilities typically found in the midwestern United States. The practice of tail docking cattle on entry into these feedlot facilities was initiated to prevent tail injuries. Tail docking is a welfare concern from the standpoint that an important method of fly avoidance is removed and the tail docking procedure is painful and often excludes local anesthesia or extended analgesia. The primary objective of this study was to describe the behavioral responses of feedlot cattle following tail docking. Thirty-six heifers were randomly assigned to 1 of 2 treatment groups: docked (DK) or control (CN). All calves received an epidural following surgical preparation of the sacrococcygeal area and postoperative intravenous flunixin meglumine. A portion of the tail of DK calves was removed using pruning shears. An elastrator band was placed near the tail tip for hemostasis and tail tips were sprayed with fly spray. IceQube accelerometers collected step counts, motion index, lying time, lying bouts, and lying bout duration during d -4 through 13. Direct observations of cattle behavior were performed on d 0, 1, and 2. Step counts of DK calves were increased (P < 0.05) on d 0, 2, 3, 4, 6, 9, 10, and 13, and motion index of DK calves was also increased (P < 0.05) on d 0, 3, 4, 9, 10, 11, and 13. Docked cattle performed rear foot stomp behavior more (P < 0.001) than CN on d 0, 1, and 2. Forty-eight hours after tail docking, DK calves had increased lying bouts per hour (1.7 vs. 0.9 on d 0; P < 0.001; 1.1 vs. 0.8 on d 1; P < 0.01) but reduced lying bout durations (12.6 vs. 47.1 min on d 0; P < 0.001; 22.6 vs. 44.7 min on d 1; P < 0.001). On d 0, DK calves twitched tails more (P < 0.05) and ruminated less (P < 0.001). Despite provision of perioperative and postoperative analgesia, we identified altered behavior in DK cattle that may reflect a compromised welfare state for tail-docked feedlot cattle. We recommend that alternative strategies to reduce tail tip injury be explored.


A new Multi-PCR-SSCP assay for simultaneous detection of isoniazid and rifampin resistance in Mycobacterium tuberculosis.
Prompt detection of drug resistance in Mycobacterium tuberculosis is essential for effective control of tuberculosis (TB). We developed a Multi-PCR-SSCP method that detects more than 80% commonly observed isoniazid (INH) and rifampin (RIF) resistance M. tuberculosis in a single assay. The usefulness of the newly developed method was evaluated with 116 clinical isolates of M. tuberculosis. Distinct SSCP patterns were observed for different mutations and the correlation between Multi-PCR-SSCP results and DNA sequencing data was strong. Using the culture-based phenotypic drug susceptibility testing as a reference, the sensitivity of the newly developed Multi-PCR-SSCP assay was determined to be 80% and 81.8% for INH and RIF, respectively. The specificity of the assay was 100% and 92%, for INH and RIF, respectively. Multi-PCR-SSCP provides a rapid and potentially more cost-effective method of detecting multidrug-resistant TB.


Nonrandom catabolism of proteins in the formation of antigenic peptide fragments.
To examine the role of protein catabolism in the formation of antigenic peptide fragments, human fibrinopeptide-immune guinea pig T cells were stimulated with the large native molecule, human fibrinogen. Two different systems were tested. In the first, we determined responses by human fibrinopeptide B (hFPB)-immune T cells, to which strain (St.) 2 guinea pigs are responders and St. 13 are nonresponders, and by human fibrinopeptide A (hFPA)-immune T cells to which St. 13 are responders and St. 2 are nonresponders. Of interest in this comparison is that both hFPA and hFPB are amino terminal peptides on the A and B chain of fibrinogen, respectively, and are readily cleaved by thrombin during fibrin formation and by other trypsin-like enzymes, leaving a carboxyl terminal Arg. Thus, if fibrinogen catabolism occurred, both antigenic peptides should be equally represented for availability in T cell responses. It was found that hFPB-immune St. 2 T cells responded to fibrinogen, but no response was observed with hPFA-immune St. 13 T cells cultured with fibrinogen. To rule out that there was a general catabolic defect in St. 13 antigen-presenting cells, fibrinogen was presented by (2 X 13)F1 macrophages to fibrinopeptide-immune parental T cells. Again it was found that F1 macrophages could present fibrinogen to hFPB-immune T cells but failed to present hFPA. In another comparison, responses with fibrinogen were also determined with des-ARg-hFPB, which lacks the carboxyl terminal Arg of hFPB, to which St. 13 are responders and St. 2 are nonresponders. The advantage of this comparison is that both antigenic determinants are contained within the same small peptide. St. 13 des-Arg-hFPB-immune T cells failed to respond in vitro by culture with human fibrinogen, suggesting that these antigenic determinants are not produced from larger peptides or proteins containing those determinants. To rule out the possibility that this was only an in vitro phenomenon, guinea pigs were immunized with the larger protein, the B chain of fibrinogen, and the immune T cells were examined for responses to fibrinopeptides derived from the B chain. Immune St. 2 T cells responded to hFPB but not to des-Arg-hFPB, whereas St. 13 T cells remained unresponsive with both peptides. These results indicate that proteolysis of larger proteins to form small antigenic peptides is not a random event and that not all potential antigenic determinants contained in a protein are produced during antigen processing.


A binding hotspot in Trypanosoma cruzi histidyl-tRNA synthetase revealed by fragment-based crystallographic cocktail screens.
American trypanosomiasis, commonly known as Chagas disease, is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. The chronic form of the infection often causes debilitating morbidity and mortality. However, the current treatment for the disease is typically inadequate owing to drug toxicity and poor efficacy, necessitating a continual effort to discover and develop new antiparasitic therapeutic agents. The structure of T. cruzi histidyl-tRNA synthetase (HisRS), a validated drug target, has previously been reported. Based on this structure and those of human cytosolic HisRS, opportunities for the development of specific inhibitors were identified. Here, efforts are reported to identify small molecules that bind to T. cruzi HisRS through fragment-based crystallographic screening in order to arrive at chemical starting points for the development of specific inhibitors. T. cruzi HisRS was soaked into 68 different cocktails from the Medical Structural Genomics of Pathogenic Protozoa (MSGPP) fragment library and diffraction data were collected to identify bound fragments after soaking. A total of 15 fragments were identified, all bound to the same site on the protein, revealing a fragment-binding hotspot adjacent to the ATP-binding pocket. On the basis of the initial hits, the design of reactive fragments targeting the hotspot which would be simultaneously covalently linked to a cysteine residue present only in trypanosomatid HisRS was initiated. Inhibition of T. cruzi HisRS was observed with the resultant reactive fragments and the anticipated binding mode was confirmed crystallographically. These results form a platform for the development of future generations of selective inhibitors for trypanosomatid HisRS.


Prognostic value of serum albumin-to-creatinine ratio in patients with acute myocardial infarction: Results from the retrospective evaluation of acute chest pain study.
The long-term association between serum albumin-to-creatinine ratio (sACR) and poor patient outcomes in acute myocardial infarction (AMI) remains unclear. This study aimed to determine whether sACR was a predictor of poor long-term survival in patients with AMI.This was a study of patients with AMI in the emergency department (ED) from the retrospective multicenter study for early evaluation of acute chest pain (REACP) study. The patients were categorized into tertiles (T1, T2, and T3) based on the admission sACR (0.445 and 0.584 g/μmol). Baseline sACR at admission to the ED was predictive of adverse outcomes. The primary outcome was all-cause mortality within the follow-up period. Cox proportional hazards models were performed to investigate the association between sACR and all-cause mortality in patients with AMI.A total of 2250 patients with AMI were enrolled, of whom 229 (10.2%) died within the median follow-up period of 10.7 (7.2-14.6) months. Patients with a lower sACR had higher all-cause mortality and adverse outcomes rates than patients with a higher sACR. Kaplan-Meier survival analysis showed that patients with a higher sACR had a higher cumulative survival rate (P < .001). Cox regression analysis showed that a decreased sACR was an independent predictor of all-cause mortality [T2 vs T1: hazard ratio (HR); 0.550, 95% confidence interval (95% CI), 0.348-0.867; P = .010 and T3 vs T1: HR, 0.305; 95% CI, 0.165-0.561; P < .001] and cardiac mortality (T2 vs T1: HR, 0.536; 95% CI, 0.332-0.866; P = .011 and T3 vs T1: HR, 0.309; 95% CI, 0.164-0.582, P < .001).The sACR at admission to ED was independently associated with adverse outcomes, indicating that baseline sACR was a useful biomarker to identify high-risk patients with AMI at an early phase in ED.


Ability of bumblebees to discriminate differences in the shape of artificial flowers of Primula sieboldii (Primulaceae).
Flower shapes are important visual cues for pollinators. However, the ability of pollinators to discriminate between flower shapes under natural conditions is poorly understood. This study focused on the diversity of flower shape in Primula sieboldii and investigated the ability of bumblebees to discriminate between flowers by combining computer graphics with a traditional behavioural experiment. Elliptic Fourier descriptors described shapes by transforming coordinate information for the contours into coefficients, and principal components analysis summarized these coefficients. Using these methods, artificial flowers were created based on the natural diversity of petal shape in P. sieboldii. Dual-choice tests were then performed to investigate the ability of the bumblebees to detect differences in the aspect ratio of petals and the depth of their head notch. The insects showed no significant ability to detect differences in the aspect ratio of the petals under natural conditions unless the morphological distance increased to an unrealistic level. These results suggest the existence of a perception threshold for distances in this parameter. The bumblebees showed a significant preference for narrow petals even after training using flowers with wide petals. The bumblebees showed a significant ability to discriminate based on the depth of the petal head notch after training using artificial flowers with a deep head notch. However, they showed no discrimination in tests with training using extreme distances between flowers in this parameter. A new type of behavioural experiment was demonstrated using real variation in flower corolla shape in P. sieboldii. If the range in aspect ratios of petals expands much further, bumblebees may learn to exhibit selective behaviour. However, because discrimination by bumblebees under natural conditions was low, there may be no strong selective behaviour based on innate or learned preferences under natural conditions.


Induction of apoptosis in nutrient-deprived cultures of hybridoma and myeloma cells.
In the present study, cell death was investigated in cultures of NS/0 myelomas and SP2/0-derived D5 hybridomas through morphological examination of cells stained with acridine orange and ethidium bromide. The relative contribution of elevated levels of lactic acid and ammonia, as well as deprivation of glutamine, cystine, and glucose on the induction of necrosis or apoptosis, was investigated. In batch culture of D5 hybridoma cells, induction of apoptotic cell death correlated with the exhaustion of glutamine, while in the case of NS/0 myelomas, it coincided with exhaustion of cystine. To determine whether limiting nutrients were the actual triggering factors for apoptosis in batch culture, exponentially growing cells were resuspended in glutamine or cystine-free media. Within 30 to 40 h, viability decreased to 50% and the nonviable cell population displayed typical apoptotic morphology, with crescents of condensed chromatin around the periphery of the nucleus, or with the entire nucleus present as one or a group of featureless, brightly staining spherical beads. Similarly, D5 hybridomas and NS/0 myelomas cultivated in glucose-free medium died mainly from apoptosis. Cells were also cultivated in fresh medium supplemented with elevated concentrations of ammonia (3.0 mM) and/or lactate (35 mM, 50 mM). This resulted in decreased viabilities and necrotic death in both cell lines. From these results, we conclude that D5 hybridomas and NS/0 myelomas deprived of essential nutrients die by apoptosis, whereas incubation in the presence of elevated levels of metabolic byproducts such as ammonia and lactate will induce necrotic cell death in these cells.


Model for eukaryotic tail-anchored protein binding based on the structure of Get3.
The Get3 ATPase directs the delivery of tail-anchored (TA) proteins to the endoplasmic reticulum (ER). TA-proteins are characterized by having a single transmembrane helix (TM) at their extreme C terminus and include many essential proteins, such as SNAREs, apoptosis factors, and protein translocation components. These proteins cannot follow the SRP-dependent co-translational pathway that typifies most integral membrane proteins; instead, post-translationally, these proteins are recognized and bound by Get3 then delivered to the ER in the ATP dependent Get pathway. To elucidate a molecular mechanism for TA protein binding by Get3 we have determined three crystal structures in apo and ADP forms from Saccharomyces cerevisae (ScGet3-apo) and Aspergillus fumigatus (AfGet3-apo and AfGet3-ADP). Using structural information, we generated mutants to confirm important interfaces and essential residues. These results point to a model of how Get3 couples ATP hydrolysis to the binding and release of TA-proteins.


Impact of Covid-19 emergency on rehabilitation services for Multiple Sclerosis: An international RIMS survey.
Covid-19 pandemic greatly impacted on the healthcare systems worldwide with negative consequences on several aspects of clinical populations. For neurological chronic conditions such as Multiple Sclerosis (MS), rehabilitation activities have been suspended or postponed during the pandemic. Rehabilitation is crucial for people with MS (PwMS) because it promotes recovery from relapses and maximizes opportunities for social participation. To better understand the impact of Covid-19 emergency on rehabilitation services for MS, the European network for rehabilitation in MS (RIMS) disseminated a survey to healthcare professionals (HPs) and representatives of the MS rehabilitation services (RSs), to explore the two different perspectives on the delivery of rehabilitation in usual circumstances and during the Covid-19 emergency. The online survey was distributed from July 9th to September 20th, 2020. Besides general information on the responders (e.g. location of center, and memebership to RIMS), information was collected on usual service delivery (e.g. settings, specialities, and types of treatment), the impact of Covid-19 circumstances (e.g. restrictions, use of personal protective equipment, and impact on work), and the use of technologiesin rehabilitation. Twenty-two representatives of MS rehabilitation services (RSs)and 143 health care professionals (HPs) responded. Most of RSs and HPs worked in services specialized for MS including a mixture of all usual rehabilitation settings (i.e. inpatient, outpatient and community setting). The majority of services adopted a multidisciplinary framework, including physical therapy, occupational therapy, social service, speech and language therapy, psychological support, dietary interventions, medical management, vocational rehabilitation and cognitive rehabilitaton. Overall, most of responders indicated they did not use technologies in their practice (e.g. for treatment or assessment). However, depending on the type of technology a low-to-medium percentage of responders declared to use some technologies before Covid-19 crisis (5-55% for RSs and 12-53% for HPs) and a low percentage planned the use after pandemic (0-14% for RSs and 1-10% for HPs). Moreover, for the responders the most feasible interventions deliverable through tele-rehabilitation were psychological support and dietary interventions, with psychological support considered the most necessary intervention to be remotely implemented. Moderate feasibility (30-60%) was reported for hands-off interventions (e.g. aerobic exercise and cognitive rehabilitation) whereas low feasibility (<30%) was reported for hands-on interventions. Feasibility was especially low when tools were used that are not adaptable at-home (e.g. hyperbaric oxygen therapy). The Covid-19 pandemic has stimulated the MS healthcare professionals to find new solutions to deliver alternative interventions to PwMS. In this context, the role of telemedicine is crucial to continue rehabilitation services at home, and limit exposure to infection. However, most of healthcare professionals have not incorporated the use of technologies. Therefore, the implementation of digital health solutions in the clinical practice needs more attention towards education on the potentials of technologies for rehabilitation and simplification of the national healthcare system reimbursement procedures for the rehabilitation technologies use.


Effect of capacitive deionization on disinfection by-product precursors.
Formation of brominated disinfection by-products (DBPs) from bromide and natural organic matter upon chlorination imposes health risks to drinking water users. In this study, capacitive deionization (CDI) was evaluated as a potential process for DBP precursor removal. Synthetic humic acid and bromide containing saline water was used as model water prior to CDI treatment. Batch experiments were conducted at cell voltages of 0.6-, 0.9-, and 1.2V to study the influence of CDI on the ratio of bromide and dissolved organic carbon, bromine substitution factor, and DBP formation potential (FP). Results showed beneficial aspects of CDI on reducing the levels of these parameters. A maximum DBPFP removal from 1510 to 1160μg/L was observed at the cell voltage of 0.6V. For the removed DBPFP, electro-adsorption played a greater role than physical adsorption. However, it is also noted that there could be electrochemical oxidations that led to reduction of humic content and formation of new dichloroacetic acid precursors at high cell voltages. Because of the potential of CDI on reducing health risks from the formation of less brominated DBPs upon subsequent chlorination, it can be considered as a potential technology for DBP control in drinking water treatment.


[Phosphine poisoning in healthcare workers].
Phosphine gas constitutes a potential and serious little-known cause of poisoning of professional nature of the medical staff and nursing care of patients who voluntarily swallow phosphides rodenticides purposes suicide. The objective of this paper is to inform to healthcare workers from urgencies, forensic and occupational health services on this occupational hazard. We present the case of a nurse who suffered from poisoning by gas phosphine confirmed through an environmental monitoring of gases in an emergency department carried out by the government service of civil protection of the State of Jalisco.


Risk and Protective Factors in Autism Spectrum Disorders: A Case Control Study in the Lebanese Population.
Autism spectrum disorders (ASD) are among the most common childhood neurodevelopmental disorders. Identification of risk and protective factors are necessary to improve the guidance of prevention and intervention strategies. Our study aims to determine the potential risk and protective factors in ASD in the Lebanese population. Our case-control study included 100 ASD patients and 100 healthy matched controls recruited from all the Lebanese districts. The data collected from the questionnaires was analyzed using SPSS 23.0. Independent Student T-test and Chi-Square test were carried out for the bivariate analysis of the data. In addition, the variables revealing a p-value < 0.05 were used for the multivariate logistic regression analysis. Multivitamins intake, especially omega 3 and vitamin B (Odds Ratio (OR) = 0.257; 95% Confidence Interval (CI) [0.115-0.579]), rich cereal diet (OR = 0.212; 95% CI [0.089-0.510]), and supplementation in iron during pregnancy (OR = 0.229; 95% CI [0.083-0.627]) were identified as protective factors against ASD. On the other hand, stress during pregnancy (OR = 6.339; 95% CI [2.845-14.125]), the presence of ASD patients in the family (OR = 7.878; 95% CI [1.877-33.065]) and the presence of attention deficit hyperactivity disorder (ADHD) patients in the family (OR = 6.981; 95% CI [1.362-35.789]) were associated with ASD. This study shed light on risk and protective factors associated with ASD in the Lebanese population. Further rigorous research, taking into consideration these factors, is needed to assist in early detection, prevention and subsequent intervention targeting ASD and its associated comorbidities, given that our study is not experimental and does not prove causality.


Investigating teacher and student effects of the Incredible Years Classroom Management Program in early elementary school.
The present paper reports on the results of a cluster randomized trial of the Incredible Years® Teacher Classroom Management Program (IY-TCM) and its effects on early elementary teachers' management strategies, classroom climate, and students' emotion regulation, attention, and academic competence. IY-TCM was implemented in 11 rural and semi-rural schools with K-2 teachers and a diverse student sample. Outcomes were compared for 45 teachers who participated in five full day training workshops and brief classroom consultation and 46 control teachers; these 91 teachers had a total of 1192 students. A high level of teacher satisfaction was found and specific aspects of the training considered most valuable for early elementary teachers were identified. Hierarchical linear modeling indicated a statistically significant intervention effect on Positive Climate in the classroom (d=0.45) that did not sustain into the next school year. No main effects on student outcomes were observed, although a priori moderator analyses indicated that students with elevated social-behavioral difficulties benefitted with regard to prosocial behavior (d=0.54) and inattention (d=-0.34). Results highlight potential benefits and limitations of a universal teacher training program for elementary students, and suggest strategies for future delivery of the IY-TCM program and areas for future research.


All-atom stability and oligomerization simulations of polyglutamine nanotubes with and without the 17-amino-acid N-terminal fragment of the Huntingtin protein.
Several neurodegenerative diseases are associated with the polyglutamine (polyQ) repeat disorder in which a segment of consecutive glutamines in the native protein is produced with too many glutamines. Huntington's disease, for example, is related to the misfolding of the Huntingtin protein which occurs when the polyQ segment has more than approximately 36 glutamines. Experimentally, it is known that the polyQ segment alone aggregates into β-rich conformations such as amyloid fibrils. Its aggregation is modulated by the number of glutamine residues as well as by the surrounding amino acid sequences such as the 17-amino-acid N-terminal fragment of Huntingtin which increases the aggregation rate. Little structural information is available, however, regarding the first steps of aggregation and the atomistic mechanisms of oligomerization are yet to be described. Following previous coarse-grained replica-exchange molecular dynamics simulations that show the spontaneous formation of a nanotube consisting of two intertwined antiparallel strands (Laghaei, R.; Mousseau, N. J. Chem. Phys. 2010, 132, 165102), we study this configuration and some extensions of it using all-atom explicit solvent MD simulations. We compare two different lengths for the polyQ segment, 40 and 30 glutamines, and we investigate the impact of the Huntingtin N-terminal residues (htt(NT)). Our results show that the dimeric nanotubes can provide a building block for the formation of longer nanotubes (hexamers and octamers). These longer nanotubes are characterized by large β-sheet propensities and a small solvent exposure of the main-chain atoms. Moreover, the oligomerization between two nanotubes occurs through the formation of protein/protein H-bonds and can result in an elongation of the water-filled core. Our results also show that the htt(NT) enhances the structural stability of the β-rich seeds, suggesting a new mechanism by which it can increase the aggregation rate of the amyloidogenic polyQ sequence.


Phospholipid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga.
The distribution of acetone-soluble (neutral glycolipid) and acetone-insoluble (phospholipid isoprenoids) lipids in oral isolates of gram-negative gliding bacteria of the genus Capnocytophaga was compared with those in a non-host-related gliding bacterium, Sporocytophaga myxococcoides. The acetone-soluble material accounted for 34 to 55% of the extracted lipids; the remainder was acetone-insoluble material. The major phospholipid was phosphatidylethanolamine (67%), with lesser amounts of lysophosphatidylethanolamine and several unidentified phosphate-containing compounds. Capnocytophaga also contained significant amounts of an ornithine-amino lipid.


The effect of a cell-to-cell communication molecule, Pseudomonas quinolone signal (PQS), produced by P. aeruginosa on other bacterial species.
One of the most important factors in the development of a bacterial community is whether the bacteria are able to grow in that habitat. The regulation of bacterial growth is generally studied in relation to physicochemical conditions, however, how bacterial communities regulate themselves remains unclear. In our previous study, it was demonstrated that a cell-to-cell communication molecule, 2-heptyl-3-hydroxy-4-quinolone, referred to as the Pseudomonas quinolone signal (PQS), affects respiring-activity in Pseudomonas aeruginosa without requiring its cognate receptor PqsR. The results suggested that PQS may affect other bacterial species, which was further examined in this study. PQS repressed the growth of several species including both Gram-negative and Gram-positive bacteria. In most cases, this effect differed from the bacteriostatic or bacteriolytic actions of antibiotics. The growth repression by PQS was inhibited when iron was added to the medium, indicating iron-chelating activity to be involved. In addition, PQS affected oxygen consumption in some species tested, and may have other underlying effects. Thus, this cell-to-cell communication molecule may influence the development of bacterial communities by regulating bacterial growth, and physicochemical factors such as iron would be important in determining its effect.


[Treatment of suppurative wounds in patients with diabetes mellitus by magnetic field and laser irradiation].
The efficacy of including a magnetic field, low-intensity laser beam, and their combination in the complex of therapeutic measures was studied in 119 patients with suppurative wounds and diabetes mellitus. With the use of magnetic field or laser beam intoxication diminished, the organism's immunological status was stabilized within a shorter time, the wound process followed a quicker course, and treatment took less time. The magnetic-laser effect has advantages over separate use of these factors.


Prevalence of foramen arcuale and its clinical significance: a meta-analysis of 55,985 subjects.
OBJECTIVE The foramen arcuale (FA) is a bony bridge located over the vertebral artery on the posterior arch of the atlas. The presence of an FA can pose a risk during neurosurgery by providing a false impression of a broader posterior arch. The aim of this study was to provide the most comprehensive investigation on the prevalence of the FA and its clinically important anatomical features. METHODS Major electronic databases were searched to identify all studies that reported relevant data on the FA and the data were pooled into a meta-analysis. RESULTS A total of 127 studies (involving 55,985 subjects) were included. The overall pooled prevalence of a complete FA was 9.1% (95% CI 8.2%-10.1%) versus an incomplete FA, which was 13.6% (95% CI 11.2%-16.2%). The complete FA was found to be most prevalent in North Americans (11.3%) and Europeans (11.2%), and least prevalent among Asians (7.5%). In males (10.4%) the complete FA was more common than in females (7.3%) but an incomplete FA was more commonly seen in females (18.5%) than in males (16.7%). In the presence of a complete FA, a contralateral FA (complete or incomplete) was found in 53.1% of cases. CONCLUSIONS Surgeons should consider the risk for the presence of an FA prior to procedures on the atlas in each patient according to sex and ethnic group. We suggest preoperative screening with computerized tomography as the gold standard for detecting the presence of an FA.


Western blot detection of scrapie-associated fibril protein in tissues outside the central nervous system from preclinical scrapie-infected mice.
We describe a method of sample preparation to detect scrapie-associated fibril (SAF) proteins in small amounts of scrapie-infected mouse tissues by Western blot analysis using an antiserum to a synthetic peptide that corresponds to the N-terminal region of hamster prion protein. SAF proteins were efficiently detected in brain tissue by this procedure. The proteins were also detected in preparations from spleen and lymph node. SAF proteins were detected in brain samples at 24 weeks after intraperitoneal infection. Using spleen samples, the proteins were detected from mice in the preclinical stage (from 4 weeks after infection), clinical symptoms of scrapie were observed in some mice from 22 weeks after infection.


Methodologic issues in terminating enrollment of a subgroup of patients in a multicenter randomized trial.
The National Emphysema Treatment Trial (NETT) was a multicenter randomized controlled trial comparing medical treatment plus lung-volume-reduction surgery (LVRS) to medical treatment alone for the treatment of severe emphysema. The primary outcomes specified for the trial were mortality from all causes and change in functional status as indicated by the change in maximum exercise capacity measured two years after randomization. A secondary objective of the trial was to define criteria to identify subgroups of patients at risk of harm or benefit from LVRS. Stopping guidelines for safety and efficacy based on 30-day mortality and a combination of overall mortality and functional status at two years were specified at the inception of the trial. Although specific subgroups of patients likely to benefit were not identified in advance, several clinical factors were specified as likely to be important in defining subgroups with differential outcome. In May 2001, with 40% of expected deaths accrued, the Data and Safety Monitoring Board determined that a subgroup of patients was at significantly higher risk of 30-day mortality from LVRS without counterbalancing evidence of functional benefit, and recommended that the protocol be modified to exclude further randomization of such patients. The trial's sponsor, the National Heart, Lung and Blood Institute, accepted the recommendation, which was rapidly communicated to participating clinics. This paper describes the operational aspects of identification of the subgroup and implementation of the recommendation to continue the trial, but to terminate enrollment of new patients in the subgroup. These aspects include notification of the investigators, the institutional review boards, the Research Group, the patients and the medical community. We also describe the repercussions of the publication and the misinterpretations of the results based on media coverage.


HMBA releases P-TEFb from HEXIM1 and 7SK snRNA via PI3K/Akt and activates HIV transcription.
Hexamethylene bisacetamide (HMBA) is a potent inducer of cell differentiation and HIV production in chronically infected cells. However, its mechanism of action remains poorly defined. In this study, we demonstrate that HMBA activates transiently the PI3K/Akt pathway, which leads to the phosphorylation of HEXIM1 and the subsequent release of active positive transcription elongation factor b (P-TEFb) from its transcriptionally inactive complex with HEXIM1 and 7SK small nuclear RNA (snRNA). As a result, P-TEFb is recruited to the HIV promoter to stimulate transcription elongation and viral production. Despite the continuous presence of HMBA, the released P-TEFb reassembles rapidly with 7SK snRNA and HEXIM1. In contrast, a mutant HEXIM1 protein that cannot be phosphorylated and released from P-TEFb and 7SK snRNA via the PI3K/Akt pathway antagonizes this HMBA-mediated induction of viral production. Thus, our studies reveal how HIV transcription is induced by HMBA and suggest how modifications in the equilibrium between active and inactive P-TEFb could contribute to cell differentiation.


Practical hyperdynamics method for systems with large changes in potential energy.
A practical hyperdynamics method is proposed to accelerate systems with highly endothermic and exothermic reactions such as hydrocarbon pyrolysis and oxidation reactions. In this method, referred to as the "adaptive hyperdynamics (AHD) method," the bias potential parameters are adaptively updated according to the change in potential energy. The approach is intensively examined for JP-10 (exo-tetrahydrodicyclopentadiene) pyrolysis simulations using the ReaxFF reactive force field. Valid boost parameter ranges are clarified as a result. It is shown that AHD can be used to model pyrolysis at temperatures as low as 1000 K while achieving a boost factor of around 10(5).


The Hekinan Children's Study: Design and Profile of Participants at Baseline.
The early life environment is now recognized as a key factor contributing to susceptibility to certain diseases in later life. We initiated a cohort study among school children in 2011 to primarily investigate the associations between lifestyle and environmental factors and some surrogate markers of chronic diseases, such as cardiometabolic risk factors (ie, obesity, high blood pressure, high blood glucose, insulin, or lipids) and cancer risk factors (ie, height and age at menarche). A baseline questionnaire asked for information, including demographic variables, medical history and use of medication, dietary habits, physical activity, sleep habits, and behavioral and emotional problems of children. Follow-up surveys are planned for the fourth grade of elementary school and the first grade of junior high school. At these follow-up surveys, fasting blood samples will be obtained to measure cardiometabolic markers. We also checked the validity of a food frequency questionnaire, which was originally created for 6-year-olds but was modified for use in older children. A total of 3,141 first-year students at elementary schools in Hekinan City, Aichi Prefecture, participated in the study. The response rate was 87.4%. The means of age and body mass index were 6.99 (standard deviation, 0.28) years and 15.3 (standard deviation, 1.7) kg/m2, respectively, in the 3,067 Japanese children (1,639 boys and 1,428 girls). This cohort will reveal determinants of cardiometabolic risk factors and cancer risk factors during childhood.


Prevalence and Determinants of Undernutrition in A Sample of Dutch Community-Dwelling Older Adults: Results from Two Online Screening Tools.
To stimulate undernutrition screening among Dutch community-dwelling adults, a website was developed with general information on healthy eating for healthy aging and self-tests. Based on cross-sectional data obtained from the self-tests, we studied nutritional risk factors (early determinants) as well as risk of undernutrition (late symptoms). SCREEN II (n = 2470) was used to asses nutritional risk factors. This tool consists of 16 items regarding nutritional intake, perception of body weight, appetite, oral health and meal preparation. An adjusted SNAQ65+ (n = 687) was used to assess risk of undernutrition. This four-item tool contains questions on weight loss, appetite, walking stairs and body mass index. Differences between age-groups (65-74, 75-84, ≥85) were tested by logistic regression. Overall prevalence of nutritional risk factors was 84.1%, and increased risk of undernutrition was 56.8%. Participants aged ≥85 scored worst on almost all items of the SCREEN II and the SNAQ65+. In conclusion: A large proportion of older adults reported early determinants for increased nutrition risk, while a smaller, yet remarkable proportion scored positive on undernutrition risk. Internet screening may be a useful, contemporary, and easy, accessible way to reach older adults who are at nutritional risk and may thus contribute to early identification and prevention of undernutrition.


Therapeutic effect of bifemelane on unilateral cerebral ischemia in gerbils.
To examine the therapeutic effect of 4-(o-benzylphenoxy)-N-methylbutylamine hydrochloride (bifemelane) on neuronal ischemia-reperfusion injury, we evaluated the neurological score in gerbils subjected to unilateral common carotid occlusion. Using the neurological score method, we selected animals which showed stroke symptoms without seizures, because seizure activity can modify the neurological outcome. Just after the clip removal following 30 min of ischemia, 0.2 ml of either vehicle (0.9% saline, n = 10) or bifemelane (25 mg/kg, n = 10) was administered intraperitoneally. Then, the neurological score and the maximum inclination at which the gerbil could maintain its equilibrium were recorded at 5, 10, and 30 min, and 1, 2, 3, 6, and 24 hr after the insult. Animals were then decapitated and their brains were processed for immunohistochemical investigation. Treatment with bifemelane after reperfusion significantly improved the neurological score and motor function. Immunohistochemistry using an antibody for microtubule-associated protein 2 clearly demonstrated preservation of staining especially in the caudate-putamen in the animals treated with bifemelane. The present findings suggest a beneficial effect of bifemelane on neuronal ischemia-reperfusion injury, which may be common after cerebral ischemia.


Role of lipids in age-related changes in the properties of muscarinic receptors in cultured rat heart myocytes.
The effects of the culture's age and of liposome treatments on the properties of muscarinic receptors in cultured rat heart myocytes prepared from the hearts of newborn (1-3 days old) rats were investigated. In these studies we investigated the binding characteristics of antagonists and agonists to the myocyte muscarinic receptors in young (5 days after plating) vs. older (14 days after plating) cultures. Our findings demonstrate that the aging of the cells in culture is accompanied by a reduction in the muscarinic binding capacity and by alterations in the proportion of high- and low-affinity states toward muscarinic agonists, as well as by striking changes in the mode of coupling of the receptors with guanine nucleotide binding protein(s) [G protein(s)]. The above effects of the culture's age occur concomitantly with alterations in the lipid composition of the cultured myocytes (in 14-day old cultures, the phosphatidylcholine/sphingomyelin ratio is reduced, and the cholesterol level is elevated). In order to explore whether the lipid composition is involved in the mechanism that alters the properties and coupling of the muscarinic receptors, we treated aging cultures with liposomes containing egg phosphatidylcholine. This treatment resulted in 14 day old cultures with a lipid composition similar to that of young cultures, and the treated myocytes demonstrated muscarinic receptor properties similar to those of young myocyte cultures. The implications for the role of membrane lipid composition and organization in determining the properties of the muscarinic receptors and their coupling with G proteins are discussed.


Two new species of shovel-jaw carp Onychostoma (Teleostei: Cyprinidae) from southern Vietnam.
Two new species of large shovel-jaw carps in the genus Onychostoma are described from the upper Krong No and middle Dong Nai drainages of the Langbiang Plateau in southern Vietnam. These new species are known from streams in montane mixed pine and evergreen forests between 140 and 1112 m. Their populations are isolated in the headwaters of the upper Sre Pok River of the Mekong basin and in the middle of the Dong Nai basin. Both species are differentiated from their congeners by a combination of the following characters: transverse mouth opening width greater than head width, 14-17 predorsal scales, caudal-peduncle length 3.9-4.2 times in SL, no barbels in adults and juveniles, a strong serrated last simple ray of the dorsal fin, and small eye diameter (20.3-21.5% HL). Onychostoma krongnoensis sp. nov. is differentiated from Onychostoma dongnaiensis sp. nov. by body depth (4.0 vs. 3.2 times in SL), predorsal scale number (14-17 vs. 14-15), dorsal-fin length (4.5 vs. 4.2 times in SL), caudal-peduncle length (3.9 vs. 4.2 times in SL), colour in life (dark vs. bright), and by mitochondrial DNA (0.2% sequence divergence). Molecular evidence indicates that both species are members of Onychostoma and are distinct from all congeners sampled (uncorrected sequence divergences at the 16S rRNA gene of >2.0% for all Onychostoma for which homologous 16S rRNA sequences are available).


Effect of a bacteriocin produced by Mycobacterium smegmatis on growth of cultured tumor and normal cells.
Growth-inhibitory effects of a partially purified bacteriocin derived from Mycobacterium smegmatis ATCC 14468 on various animal cells transformed by tumor viruses, human malignant cells, and normal cells in the same species were studied. A growth-inhibitory effect of the bacteriocin on these cultured cells was determined by counting the residual cells. The bacteriocin inhibited virally transformed animal cells (mKS-A TU-7, 155-4 T2, and XC cells) and human malignant cells (AS-II and HGC-27 cells). The inhibitory effect increased with an increase in the bacteriocin activity. The bacteriocin sensitivities of transformed animal cells were relatively higher than were those of human malignant cells, while normal cells in the same species were practically insensitive to the bacteriocin. Differences in the degree of bacteriocin sensitivity were observed among tumor cell lines. Simian virus (SV) 40-transformed hamster cells (TSV-5 cells), which grow rapidly, were less sensitive to the bacteriocin. The cell membrane of SV40-transformed BALB/c mouse cells (mKS-A TU-7 cells) adsorbed the bacteriocin much more than did the cell membrane of nontransformed BALB/3T3 cells. The results seem to indicate that the inhibitory effect of bacteriocin 14468 on cultured mammalian cells probably depends on the binding sites for the bacteriocin which appear or increase by malignant transformation on cytoplasmic membrane.


Fluorescence polarization immunoassay for zidovudine.
We report a fluorescence polarization immunoassay (FPIA) for zidovudine (azidothymidine; Retrovir). This assay is accurate and specific over the clinically relevant range of zidovudine concentrations in serum (from 1 to 1,250 ng/ml; from 0.004 to 4.8 microM) and is unaffected by potentially interfering compounds in the sera of patients with renal or hepatic failure. Cross-reactivity with structural analogs of zidovudine (including zidovudine glucuronide) is less than 0.05%, except for cross-reactivities of 0.2, 0.3, and 0.4% with 3-methylthymidine, 3',5'-dideoxythymidine, and A22U (the optical isomer of zidovudine), respectively. The FPIA for zidovudine is more sensitive and more specific than high-performance liquid chromatography (HPLC); it requires 50 to 60 or 200 versus 500 microliters of serum and is faster to perform (45 specimens per h with the FPIA versus 3 specimens per h with HPLC). The zidovudine FPIA compares well with the radioimmunoassay. A correlation coefficient of 0.992 was observed with 31 serum specimens examined by both methods. All three assays (FPIA, radioimmunoassay, and HPLC) are unaffected by the heat treatment used to inactivate human immunodeficiency virus. The zidovudine FPIA should be particularly useful for analyzing specimens from large numbers of human immunodeficiency virus-infected patients receiving zidovudine in current clinical trials.


Human mesenchymal stem cells suppress donor CD4(+) T cell proliferation and reduce pathology in a humanized mouse model of acute graft-versus-host disease.
Acute graft-versus-host disease (aGVHD) is a life-threatening complication following allogeneic haematopoietic stem cell transplantation (HSCT), occurring in up to 30-50% of patients who receive human leucocyte antigen (HLA)-matched sibling transplants. Current therapies for steroid refractory aGVHD are limited, with the prognosis of patients suboptimal. Mesenchymal stem or stromal cells (MSC), a heterogeneous cell population present in many tissues, display potent immunomodulatory abilities. Autologous and allogeneic ex-vivo expanded human MSC have been utilized to treat aGVHD with promising results, but the mechanisms of therapeutic action remain unclear. Here a robust humanized mouse model of aGVHD based on delivery of human peripheral blood mononuclear cells (PBMC) to non-obese diabetic (NOD)-severe combined immunodeficient (SCID) interleukin (IL)-2rγ(null) (NSG) mice was developed that allowed the exploration of the role of MSC in cell therapy. MSC therapy resulted in the reduction of liver and gut pathology and significantly increased survival. Protection was dependent upon the timing of MSC therapy, with conventional MSC proving effective only after delayed administration. In contrast, interferon (IFN)-γ-stimulated MSC were effective when delivered with PBMC. The beneficial effect of MSC therapy in this model was not due to the inhibition of donor PBMC chimerism, as CD45(+) and T cells engrafted successfully in this model. MSC therapy did not induce donor T cell anergy, FoxP3(+) T regulatory cells or cause PBMC apoptosis in this model; however, it was associated with the direct inhibition of donor CD4(+) T cell proliferation and reduction of human tumour necrosis factor-α in serum.


Capacitation of human naïve pluripotent stem cells for multi-lineage differentiation.
Human naïve pluripotent stem cells (PSCs) share features with the pre-implantation epiblast. They therefore provide an unmatched opportunity for characterising the developmental programme of pluripotency in Homo sapiens Here, we confirm that naïve PSCs do not respond directly to germ layer induction, but must first acquire competence. Capacitation for multi-lineage differentiation occurs without exogenous growth factor stimulation and is facilitated by inhibition of Wnt signalling. Whole-transcriptome profiling during this formative transition highlights dynamic changes in gene expression, which affect many cellular properties including metabolism and epithelial features. Notably, naïve pluripotency factors are exchanged for postimplantation factors, but competent cells remain devoid of lineage-specific transcription. The gradual pace of transition for human naïve PSCs is consistent with the timespan of primate development from blastocyst to gastrulation. Transcriptome trajectory during in vitro capacitation of human naïve cells tracks the progression of the epiblast during embryogenesis in Macaca fascicularis, but shows greater divergence from mouse development. Thus, the formative transition of naïve PSCs in a simple culture system may recapitulate essential and specific features of pluripotency dynamics during an inaccessible period of human embryogenesis.


Family dinner and diet quality among older children and adolescents.
The proportion of children eating dinner with their families declines with age and has decreased over time. Few data exist concerning the nutritional effect of eating family dinner. To examine the associations between frequency of eating dinner with family and measures of diet quality. Cross-sectional. A national convenience sample. There were 8677 girls and 7525 boys in the study, aged 9 to 14 years, who were children of the participants in the ongoing Nurses' Health Study II. We collected data from a self-administered mailed survey, including food and nutrient intakes from a validated semiquantitative food frequency questionnaire. Main outcome measures included servings per day of selected foods and food groups, daily intakes of selected macronutrients and micronutrients, and frequency of multivitamin use. Approximately 17% of participants ate dinner with members of their family never or some days, 40% on most days, and 43% every day. More than half of the 9-year-olds ate family dinner every day, whereas only about one third of 14-year-olds did so. In age- and sex-adjusted logistic regression models, the odds ratios associated with a frequency of family dinner of most days compared with never or some days, or every day compared with most days, were as follows: for eating at least 5 servings per day of fruits and vegetables, 1.45 (95% confidence interval [CI], 1.37-1.53); for eating any fried foods away from home, 0.67 (95% CI, 0.64-0.70); and for drinking any soda, 0.73 (95% CI, 0.66-0.80). Multiple linear regression showed that an increased frequency of family dinner was also associated with substantially higher intake of several nutrients, including fiber, calcium, folate, iron, vitamins B6, B12, C, and E; lower glycemic load; and lower intake of saturated and trans fat as a percentage of energy. We observed little or no effect on intakes of whole dairy products, red meat, or snack foods. Patterns were similar for boys and girls. Eating family dinner was associated with healthful dietary intake patterns, including more fruits and vegetables, less fried food and soda, less saturated and trans fat, lower glycemic load, more fiber and micronutrients from food, and no material differences in red meat or snack foods.


Network dynamics of eukaryotic LTR retroelements beyond phylogenetic trees.
Sequencing projects have allowed diverse retroviruses and LTR retrotransposons from different eukaryotic organisms to be characterized. It is known that retroviruses and other retro-transcribing viruses evolve from LTR retrotransposons and that this whole system clusters into five families: Ty3/Gypsy, Retroviridae, Ty1/Copia, Bel/Pao and Caulimoviridae. Phylogenetic analyses usually show that these split into multiple distinct lineages but what is yet to be understood is how deep evolution occurred in this system. We combined phylogenetic and graph analyses to investigate the history of LTR retroelements both as a tree and as a network. We used 268 non-redundant LTR retroelements, many of them introduced for the first time in this work, to elucidate all possible LTR retroelement phylogenetic patterns. These were superimposed over the tree of eukaryotes to investigate the dynamics of the system, at distinct evolutionary times. Next, we investigated phenotypic features such as duplication and variability of amino acid motifs, and several differences in genomic ORF organization. Using this information we characterized eight reticulate evolution markers to construct phenotypic network models. The evolutionary history of LTR retroelements can be traced as a time-evolving network that depends on phylogenetic patterns, epigenetic host-factors and phenotypic plasticity. The Ty1/Copia and the Ty3/Gypsy families represent the oldest patterns in this network that we found mimics eukaryotic macroevolution. The emergence of the Bel/Pao, Retroviridae and Caulimoviridae families in this network can be related with distinct inflations of the Ty3/Gypsy family, at distinct evolutionary times. This suggests that Ty3/Gypsy ancestors diversified much more than their Ty1/Copia counterparts, at distinct geological eras. Consistent with the principle of preferential attachment, the connectivities among phenotypic markers, taken as network-represented combinations, are power-law distributed. This evidences an inflationary mode of evolution where the system diversity; 1) expands continuously alternating vertical and gradual processes of phylogenetic divergence with episodes of modular, saltatory and reticulate evolution; 2) is governed by the intrinsic capability of distinct LTR retroelement host-communities to self-organize their phenotypes according to emergent laws characteristic of complex systems. This article was reviewed by Eugene V. Koonin, Eric Bapteste, and Enmanuelle Lerat (nominated by King Jordan).


Concurrent reinforcement and alcohol: interactive effects on human aggressive behavior.
The relationship between alcohol and human aggressive behavior was studied under conditions in which nonaggressive response requirements maintained by point presentation were manipulated. Six normal adult male recreational alcohol users pressed buttons that produced points (redeemable for money) on their own counters or ostensibly subtracted points (money) from another fictitious subject described as participating in the same study at another location. During random components of each session, subjects were required to press a button 50, 200 or 500 times in order to produce point increments on their own counters. Ten responses on a second button were required to subtract a point from the other subject during all components. Aggressive responding was engendered by occasional point subtractions which were attributed to the fictitious subject. Alcohol (0, 0.125, 0.25 and 0.5 g/kg of 95% ethanol) produced dose-related increases in aggressive responding. Aggressive responding was also significantly related to the concurrent schedule of point presentation. Finally, a significant interaction between alcohol and concurrent point presentation contingencies was observed. These results indicate that aggressive responding is affected by ongoing reinforcement contingencies not directly related to aggressive behavior and that reinforcement schedule interaction may be one factor mediating the situation-dependent nature of alcohol's effects on aggressive behavior.


Intensive Care Unit Utilization Following Major Surgery and the Nurse Work Environment.
Across hospitals, there is wide variation in ICU utilization after surgery. However, it is unknown whether and to what extent the nurse work environment is associated with a patient's odds of admission to an intensive care unit. To estimate the relationship between hospitals' nurse work environment and a patient's likelihood of ICU admission and mortality following surgery. A cross-sectional study of 269 764 adult surgical patients in 453 hospitals was conducted. Logistic regression models were used to estimate the effects of the work environment on the odds of patients' admission to the intensive care unit and mortality. Patients in hospitals with good work environments had 16% lower odds of intensive care unit admission and 15% lower odds of mortality or intensive care unit admission than patients in hospitals with mixed or poor environments. Patients in hospitals with better nurse work environments were less likely to be admitted to an intensive care unit and less likely to die. Hospitals with better nurse work environments may be better equipped to provide postoperative patient care on lower acuity units.


Expression of Tiam-1 in the developing brain suggests a role for the Tiam-1-Rac signaling pathway in cell migration and neurite outgrowth.
During development proper neuronal migration and neurite extension are essential for the formation of functional neuronal networks. These processes require the reorganization of the cytoskeleton by modifying the dynamics of actin filaments and microtubules. The Rho subfamily of GTPases regulates actin cytoskeletal changes during development. Tiam-1, a GDP-GTP exchange factor for the small GTPase Rac and implicated in tumor invasion and metastasis, is expressed in the developing CNS. To study the function of Tiam-1 in neuronal migration and neurite extension, we examined the pattern of Tiam-1 expression in weaver mice, in which cerebellar granule cells fail to migrate to their final position and subsequently die. Tiam-1 is expressed in wild-type granule cells as they migrate to the internal granular layer and send axone. In contrast, weaver homozygous animals do not express. Tiam-1 in premigratory granule cells. Heterozygous animals, in which granule cells exhibit a slow rate of migration, express low levels of Tiam-1. In the cerebral cortex, Tiam-1 is also expressed in migrating neurons. Our findings suggest that Tiam-1 contributes to cytoskeletal reorganization required during cell migration and neurite extension in defined neuronal populations, presumably by activation of Rac.


Learning from the "tail end" of de-implementation: the case of chemical castration for localized prostate cancer.
Men with prostate cancer are often treated with the suppression of testosterone through long-acting injectable drugs termed chemical castration or androgen deprivation therapy (ADT). In most cases, ADT is not an appropriate treatment for localized prostate cancer, indicating low-value care. Guided by the Theoretical Domains Framework (TDF) and the Behavior Change Wheel's Capability, Opportunity, Motivation Model (COM-B), we conducted a qualitative study to identify behavioral determinants of low-value ADT use to manage localized prostate cancer, and theory-based opportunities for de-implementation strategy development. We used national cancer registry and administrative data from 2016 to 2017 to examine the variation in low-value ADT use across Veterans Health Administration facilities. Using purposive sampling, we selected high- and low-performing sites to conduct 20 urology provider interviews regarding low-value ADT. We coded transcripts into TDF domains and mapped content to the COM-B model to generate a conceptual framework for addressing low-value ADT practices. Our interview findings reflected provider perspectives on prescribing ADT as low-value localized prostate cancer treatment, including barriers and facilitators to de-implementing low-value ADT. We characterized providers as belonging in 1 of 3 categories with respect to low-value ADT use: 1) never prescribe 2); willing, under some circumstances, to prescribe: and 3) prescribe as an acceptable treatment option. Provider capability to prescribe low-value ADT depended on their knowledge of localized prostate cancer treatment options (knowledge) coupled with interpersonal skills to engage patients in educational discussion (skills). Provider opportunity to prescribe low-value ADT centered on the environmental resources to inform ADT decisions (e.g., multi-disciplinary review), perceived guideline availability, and social roles and influences regarding ADT practices, such as prior training. Provider motivation involved goals of ADT use, including patient preferences, beliefs in capabilities/professional confidence, and beliefs about the consequences of prescribing or not prescribing ADT. Use of the TDF domains and the COM-B model enabled us to conceptualize provider behavior with respect to low-value ADT use and clarify possible areas for intervention to effect de-implementation of low-value ADT prescribing in localized prostate cancer. ClinicalTrials.gov , NCT03579680.


Unusual spectroscopic and ligand binding properties of the cytochrome P450-flavodoxin fusion enzyme XplA.
The Rhodococcus rhodochrous strain 11Y XplA enzyme is an unusual cytochrome P450-flavodoxin fusion enzyme that catalyzes reductive denitration of the explosive hexahydro-1,3,5-trinitro-1,3,5-triazene (RDX). We show by light scattering that XplA is a monomeric enzyme. XplA has high affinity for imidazole (K(d) = 1.6 μM), explaining previous reports of a red-shifted XplA Soret band in pure enzyme. The true Soret maximum of XplA is at 417 nm. Similarly, unusually weak XplA flavodoxin FMN binding (K(d) = 1.09 μM) necessitates its purification in the presence of the cofactor to produce hallmark flavin contributions absent in previously reported spectra. Structural and ligand-binding data reveal a constricted active site able to accommodate RDX and small inhibitory ligands (e.g. 4-phenylimidazole and morpholine) while discriminating against larger azole drugs. The crystal structure also identifies a high affinity imidazole binding site, consistent with its low K(d), and shows active site penetration by PEG, perhaps indicative of an evolutionary lipid-metabolizing function for XplA. EPR studies indicate heterogeneity in binding mode for RDX and other ligands. The substrate analog trinitrobenzene does not induce a substrate-like type I optical shift but creates a unique low spin EPR spectrum due to influence on structure around the distal water heme ligand. The substrate-free heme iron potential (-268 mV versus NHE) is positive for a low spin P450, and the elevated potential of the FMN semiquinone/hydroquinone couple (-172 mV) is also an adaptation that may reflect (along with the absence of a key Thr/Ser residue conserved in oxygen-activating P450s) the evolution of XplA as a specialized RDX reductase catalyst.


[Changes to Dutch College of General Practitioners guideline 'Micturition problems in men'].
Major disadvantages are associated with the early diagnostics of prostate cancer; for this reason routine determination of PSA level is not recommended in the Netherlands. If, after guidance, the patient still wants further investigations and rectal examination is not abnormal, the Dutch College of General Practitioners guideline 'Micturition problems in men' recommends that a PSA test should be done. The cut-off PSA level for referring patients for secondary care has been lowered from 4 ng/ml to 3 ng/ml, meaning that this guideline is now in accordance with the Dutch Society for Urology's multidisciplinary guideline "Prostate Carcinoma". Using the Prostate Cancer Risk Calculator (Prostaatwijzer-3), the PSA level and ultrasound-guided measurement of prostate volume can be used to calculate the risk of prostate cancer. The risk calculated is then used a starting point in determining if invasive follow-up diagnostic tests should be implemented.


Landscape and Clinical Significance of Immune Checkpoint in Cutaneous Melanoma.
The incidence of cutaneous melanoma (CM) is increasing, and its prognosis is not optimistic. Although immune checkpoint (ICP) inhibitors are effective in the treatment of CM patients, they are not effective for all CM patients. There is an urgent need for a marker to predict both the prognosis and the immunotherapy effect in patients with CM. Two groups of patients with greatly different prognosis and response to immunotherapy were identified by unwatched cluster exploration of TCGA on the basis of 34 ICPs. The prognosis and immunotherapy effect of CM were predicted by developing a precise and given signature on the basis of ICPs, and a multivariate Cox risk regression model was established from the TCGA cohort consisting of 454 CM samples. The model was validated in 210 and 231 samples in the test and verification cohorts, respectively. The prognosis in clinical subgroups was predicted by the classification system. High-risk patients had poorer responses to chemotherapy and immunotherapy. Finally, the signature was recognized as an independent prognostic factor. Based on checkpoint-based signature (ICPBS) and clinical characteristics, we constructed a nomogram for the prognosis in patients with CM, which was superior to ICPBS in efficacy than ICPBS alone. As a useful prognostic tool to further improve cancer immunotherapy, the signature can accurately predict recurrence and overall survival among patients with CM.


Minocycline Alleviates Cluster Formation of Activated Microglia and Age-dependent Dopaminergic Cell Death in the Substantia Nigra of Zitter Mutant Rat.
Microglial activation is a component of neurodegenerative pathology. Here, we examine whether activated microglia participate in age-related dopaminergic (DA) cell death in the substantia nigra pars compacta (SNc) of the zitter (zi/zi) rat, a mutant characterized by deletion of the attractin gene. Confocal microscopy with double-immunohistochemical staining revealed activated microglia-formed cell-clusters surrounding DA neurons in the SNc from 2 weeks after birth. An immunoelectron microscopic study showed that the cytoplasm of activated microglia usually contains phagosome-like vacuoles and lamellar inclusions. Expression levels of the pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) were increased in the midbrain of 2-month-old zi/zi rats. Chronic treatment with the anti-inflammatory agent minocycline altered the morphology of the microglia, reduced cluster formation by the microglia, and attenuated DA cell death in the SNc, and reduced the expression of IL-1β in the midbrain. These results indicate that activated microglia, at least in part and especially at the initial phase, contribute to DA cell death in the SNc of the zi/zi rat.


Primed adaptation tolerates extensive structural and size variations of the CRISPR RNA guide in Haloarcula hispanica.
Recent studies on CRISPR adaptation revealed that priming is a major pathway of spacer acquisition, at least for the most prevalent type I systems. Priming is guided by a CRISPR RNA which fully/partially matches the invader DNA, but the plasticity of this RNA guide has not yet been characterized. In this study, we extensively modified the two conserved handles of a priming crRNA in Haloarcula hispanica, and altered the size of its central spacer part. Interestingly, priming is insusceptible to the full deletion of 3' handle, which seriously impaired crRNA stability and interference effects. With 3' handle deletion, further truncation of 5' handle revealed that its spacer-proximal 6 nucleotides could provide the least conserved sequence required for priming. Subsequent scanning mutation further identified critical nucleotides within 5' handle. Besides, priming was also shown to tolerate a wider size variation of the spacer part, compared to interference. These data collectively illustrate the high tolerance of priming to extensive structural/size variations of the crRNA guide, which highlights the structural flexibility of the crRNA-effector ribonucleoprotein complex. The observed high priming effectiveness suggests that primed adaptation promotes clearance of the fast-replicating and ever-evolving viral DNA, by rapidly and persistently multiplexing the interference pathway.


Artificial muscle for end-stage heart failure.
We describe a device made of artificial muscle for the treatment of end-stage heart failure as an alternative to current heart assist devices. The key component is a matrix of nitinol wires and aramidic fibers called Biometal muscle (BM). When heated electrically, it produces a motorless, smooth, and lifelike motion. The BM is connected to a carbon fiber scaffold, tightening the heart and providing simultaneous assistance to the left and right ventricles. A pacemaker-like microprocessor drives the contraction of the BM. We tested the device in a dedicated bench model of diseased heart. It generated a systolic pressure of 75 mm Hg and ejected a maximum of 330 ml/min, with an ejection fraction of 12%. The device required a power supply of 6 V, 250 mA. This could be the beginning of an era in which BMs integrate or replace the mechanical function of natural muscles.


UXT chaperone prevents proteotoxicity by acting as an autophagy adaptor for p62-dependent aggrephagy.
p62/SQSTM1 is known to act as a key mediator in the selective autophagy of protein aggregates, or aggrephagy, by steering ubiquitinated protein aggregates towards the autophagy pathway. Here, we use a yeast two-hybrid screen to identify the prefoldin-like chaperone UXT as an interacting protein of p62. We show that UXT can bind to protein aggregates as well as the LB domain of p62, and, possibly by forming an oligomer, increase p62 clustering for its efficient targeting to protein aggregates, thereby promoting the formation of the p62 body and clearance of its cargo via autophagy. We also find that ectopic expression of human UXT delays SOD1(A4V)-induced degeneration of motor neurons in a Xenopus model system, and that specific disruption of the interaction between UXT and p62 suppresses UXT-mediated protection. Together, these results indicate that UXT functions as an autophagy adaptor of p62-dependent aggrephagy. Furthermore, our study illustrates a cooperative relationship between molecular chaperones and the aggrephagy machinery that efficiently removes misfolded protein aggregates.


FOXA1 promotes proliferation, migration and invasion by transcriptional activating KRT7 in human gastric cancer cells.
Gastric cancer (GC) is the fourth leading cause of gynecological malignancies worldwide. 5-fluorouracil (5-FU)-mediated chemotherapy is the adjuvant treatment for patients with GC following surgical resection. Many studies have indicated the cancer-type specific roles of forkhead box protein A1 (FOXA1) and keratin 7 (KRT7) in human malignancies. However, the potential mechanism underlying the involvement of FOXA1 and KRT7 in the pathogenesis and chemoresistance of GC are still not entirely clear. In our study, gain- and loss-of-function experiments proved that FOXA1 promoted cell proliferation, migration and invasion in AGS and SGC-7901 cells. Consequently, KRT7 was identified to be transcriptional activated by FOXA1 using Dual luciferase reporter assay. Our results also indicated that FOXA1 exerted its functions in enhancing viability and invasion of AGS and SGC-7901 cells through activating KRT7. Finally, interference of FOXA1 or KRT7 increased the chemosensitivity of AGS and SGC-7901 cells to 5-fluorouracil (5-Fu) treatment by suppressing cell proliferation. In conclusion, these data indicate that FOXA1 promoted proliferation, migration, invasion, and decreased chemosensitivity of GC cells to 5-Fu treatment through transcriptional activator KRT7. The present study provides a novel therapeutic strategy for the enhancement of efficacy in GC treatment and provides important insights into the molecular mechanism underlying 5-FU-mediated chemoresistance.


Regular Khat (Catha edulis) chewing is associated with elevated diastolic blood pressure among adults in Butajira, Ethiopia: a comparative study.
Fresh leaves and buds of the Khat plant (Catha edulis) contain Cathinone, an amphetamine like alkaloid responsible for its pharmacological action. Chewing of Khat has been associated with a transient rise in blood pressure and heart rate in experimental studies. Few studies examined the effect of regular or frequent Khat chewing on blood pressure at the population level. This study was conducted to examine the association of regular Khat chewing with blood pressure among adults. We compared systolic and diastolic blood pressure of adults 35-65 years of age who reported regular chewing of Khat during the preceding five years to those who never chewed Khat during the same period. Study participants were recruited from purposively selected urban and rural villages of Butajira District in Ethiopia. The comparative groups, chewers (334) and non-chewers (330), were identified from among the general population through a house-to-house visit using a screening questionnaire. They were frequency-matched for sex and age within a five-year range. Data were collected through structured interviews and physical measurements including blood pressure, weight and height. The prevalence of hypertension was significantly higher among Khat chewers (13.4%) than non-chewers (10.7%), odds ratio (OR) = 1.66 (95% confidence interval (CI) 1.05, 3.13). A considerably high proportion of chewers (29.9%) than non-chewers (20.6%) had sub-optimal diastolic blood pressure (> 80 mmHg). The mean (sd) diastolic blood pressure was significantly higher among Khat chewers [75.0 (11.6)] than non-chewers [72.9 (11.7)], P < 0.05. Similarly, Khat chewers had significantly higher mean (sd) heart rate [76.3 (11.5)] than non-chewers [73.9 (12.6)], P < 0.05. There was no significant difference in mean systolic blood pressure between the two groups. Regular chewing of Khat is associated with elevated mean diastolic blood pressure, which is consistent with the peripheral vasoconstrictor effect of Cathinone. Regular Khat chewing may have sustained effects on the cardiovascular system that can contribute to elevated blood pressure at the population level.


Recovery of corneal innervation following photorefractive keratoablation.
To study the morphological recovery of rabbit corneal nerves for up to 12 months after photorefractive keratoablation. The histochemical acetylcholinesterase reaction was used to demonstrate morphological features in rabbit corneal nerves at 1 hour, at 6 weeks, and at 3, 6, or 12 months after photorefractive keratoablation. Photorefractive keratoablation ablated the subepithelial nerve plexus, leaving sharply cut stromal nerves. Epithelial innervation was almost completely restored in 3 months, with innervation proceeding from the epithelial nerve plexuses at the wound margins, but the stromal nerves showed abnormal morphological features throughout the study. Abnormally coiled, regenerating axons emerged from the cut stromal nerves. The first of them were observed to penetrate into the epithelium 3 months postoperatively. The acetylcholinesterase reaction of the epithelial cell membranes was weaker in the wound area, except in the specimens that were observed at 12 months. Epithelial innervation was restored relatively quickly; otherwise, neural recovery was slow. The structure and architecture of the stromal nerves was abnormal even in the the specimens that were observed at 12 months. We hypothesize that corneal nerve damage induced by deep photoablations may correlate with the predictability problems and haze.


An Eight-year, Single-center Experience on Ultrasound Assisted Thrombolysis with Moderate-dose, Slow-infusion Regimen in Pulmonary Embolism.
There is limited data on moderate-dose with slow-infusion thrombolytic regimen by ultrasound-asssisted-thrombolysis (USAT) in patients with acute pulmonary embolism (PE). In this study, our eight-year experience on USAT with moderate-dose, slow-infusion tissue-type plasminogen activator (t-PA) regimen in patients with PE at intermediate-high- and high-risk was presented, and short-, and long-term effectiveness and safety outcomes were evaluated. Our study is based on the retrospective evaluation of 225 patients with PE having multiple comorbidities who underwent USAT. High- and intermediate-high-risk were noted in 14.7% and in 85.3% of patients, respectively. Mean t-PA dosage was 35.4±13.3 mg, and the infusion duration was 26.6±7.7 h. Measures of pulmonary artery (PA) obstruction and right ventricle (RV) dysfunction were improved within days (p<0.0001 for all). During the hospital stay, major and minor bleeding and mortality rates were 6.2%, 12.4%, and 6.2%, respectively. Bleeding and unresolved PE accounted for 50% and 42.8% of in-hospital mortality, respectively. Age, rate, and duration of t-PA were not associated with in-hospital major bleeding and mortality. Oxygen saturation exceeded 90% in 91.2% of patients at discharge. During follow-up of median 962 (610-1894) days, high-risk status related to 30-day mortality, whereas age >65 years was associated with long-term mortality. Our real-life experience with USAT with moderate-dose, slow-infusion t-PA regimen in patients with PE at high-and intermediate-high risk demonstrated clinically relevant improvements in PA obstructive burden and RV dysfunction. Age, rate or infusion duration of t-PA was not related to major bleeding or mortality risk, whereas unresolved obstruction remained as a lethal issue.


International Pediatric COVID-19 Severity Over the Course of the Pandemic.
Multiple SARS-CoV-2 variants have emerged over the COVID-19 pandemic. The implications for COVID-19 severity in children worldwide are unclear. To determine whether the dominant circulating SARS-CoV-2 variants of concern (VOCs) were associated with differences in COVID-19 severity among hospitalized children. Clinical data from hospitalized children and adolescents (younger than 18 years) who were SARS-CoV-2 positive were obtained from 9 countries (Australia, Brazil, Italy, Portugal, South Africa, Switzerland, Thailand, UK, and the US) during 3 different time frames. Time frames 1 (T1), 2 (T2), and 3 (T3) were defined to represent periods of dominance by the ancestral virus, pre-Omicron VOCs, and Omicron, respectively. Age groups for analysis were younger than 6 months, 6 months to younger than 5 years, and 5 to younger than 18 years. Children with an incidental positive test result for SARS-CoV-2 were excluded. SARS-CoV-2 hospitalization during the stipulated time frame. The severity of disease was assessed by admission to intensive care unit (ICU), the need for ventilatory support, or oxygen therapy. Among 31 785 hospitalized children and adolescents, the median age was 4 (IQR 1-12) years and 16 639 were male (52.3%). In children younger than 5 years, across successive SARS-CoV-2 waves, there was a reduction in ICU admission (T3 vs T1: risk ratio [RR], 0.56; 95% CI, 0.42-0.75 [younger than 6 months]; RR, 0.61, 95% CI; 0.47-0.79 [6 months to younger than 5 years]), but not ventilatory support or oxygen therapy. In contrast, ICU admission (T3 vs T1: RR, 0.39, 95% CI, 0.32-0.48), ventilatory support (T3 vs T1: RR, 0.37; 95% CI, 0.27-0.51), and oxygen therapy (T3 vs T1: RR, 0.47; 95% CI, 0.32-0.70) decreased across SARS-CoV-2 waves in children 5 years to younger than 18 years old. The results were consistent when data were restricted to unvaccinated children. This study provides valuable insights into the impact of SARS-CoV-2 VOCs on the severity of COVID-19 in hospitalized children across different age groups and countries, suggesting that while ICU admissions decreased across the pandemic in all age groups, ventilatory and oxygen support generally did not decrease over time in children aged younger than 5 years. These findings highlight the importance of considering different pediatric age groups when assessing disease severity in COVID-19.


Design, development, and validation of a strand-specific RT-qPCR assay for GI and GII human Noroviruses.
Human noroviruses (HuNoV) are the major cause of viral gastroenteritis worldwide. Similar to other positive-sense single-stranded RNA viruses, norovirus RNA replication requires the formation of a negative strand RNA intermediate. Methods for detecting and quantifying the viral positive or negative sense RNA in infected cells and tissues can be used as important tools in dissecting virus replication. In this study, we have established a sensitive and strand-specific Taqman-based quantitative polymerase chain reaction (qPCR) assay for both genogroups GI and GII HuNoV. This assay shows good reproducibility, has a broad dynamic range and is able to detect a diverse range of isolates. We used tagged primers containing a non-viral sequence for the reverse transcription (RT) reaction and targeted this tag in the succeeding qPCR reaction to achieve strand specificity. The specificity of the assay was confirmed by the detection of specific viral RNA strands in the presence of high levels of the opposing strands, in both RT and qPCR reactions. Finally, we further validated the assay in norovirus replicon-bearing cell lines and norovirus-infected human small intestinal organoids, in the presence or absence of small-molecule inhibitors. Overall, we have established a strand-specific qPCR assay that can be used as a reliable method to understand the molecular details of the human norovirus life cycle.


Diagnostic and therapeutic endoscopy.
Within the gastrointestinal tract, there is no shorter segment with a higher cancer incidence than the gastro-esophageal junction. Flexible endoscopy is the mainstay for diagnosis of early and advanced esophageal adenocarcinoma, for the treatment of early lesions, and also for the palliation of advanced cancer. New developments in endoscopy aim to improve the diagnosis and treatment of esophageal cancer. These include high resolution and magnification endoscopy in combination with chromendoscopy, and techniques based on modulation of the features of light bundles, such as narrow band imaging, fluorescence endoscopy, and elastic scattering spectroscopy. The value of these techniques for the surveillance of distal esophageal neoplasia needs further study. Furthermore, new methods of tissue sampling and evaluation are being studied to augment identification and staging of patients at risk for cancer. Finally, newer instruments may decrease patient burden during endoscopy, making screening and surveillance more acceptable from a patient's perspective. This review discusses the new developments in flexible endoscopy for diagnosis and therapy of early and advanced and advanced esophageal adenocarcinoma.


Long-term trends in the incidence and management of shoulder dystocia in a tertiary obstetric unit in Hong Kong.
Because there have been changes in the management of macrosomic pregnancies and shoulder dystocia in the past decade, this study was conducted to compare the incidences of shoulder dystocia and perinatal outcomes between the periods of 2000-2009 and 2010-2019. This retrospective study was conducted in a tertiary obstetric unit. All cases of shoulder dystocia were identified using the hospital's electronic database. The incidences, maternal and fetal characteristics, obstetric management methods, and perinatal outcomes were compared between the two study periods. The overall incidence of shoulder dystocia decreased from 0.23% (134/58 326) in 2000-2009 to 0.16% (108/65 683) in 2010-2019 (P=0.009), mainly because of the overall decline in the proportion of babies with macrosomia (from 3.3% to 2.3%; P<0.001). The improved success rates of the McRoberts' manoeuvre (from 31.3% to 47.2%; P=0.012) and posterior arm extraction (from 52.9% to 92.3%; P=0.042) allowed a greater proportion of affected babies to be delivered within 2 minutes (from 59.0% to 79.6%; P=0.003). These changes led to a significant reduction in the proportion of fetuses with low Apgar scores: <5 at 1 minute of life (from 13.4% to 5.6%; P=0.042) and <7 at 5 minutes of life (from 11.9% to 4.6%; P=0.045). More proactive management of macrosomic pregnancies and enhanced training in the acute management of shoulder dystocia led to significant improvements in shoulder dystocia incidence and perinatal outcomes from 2000-2009 to 2010-2019.


Noneffect of rifampin on theophylline disposition in rabbits.
The pharmacokinetic disposition of theophylline after a single 10mg/kg intravenous dose of aminophylline was evaluated in six rabbits. Three then received rifampin (50mg/kg) a day for 14 days and the disposition of theophylline was re-evaluated in all six rabbits after another 10mg/kg aminophylline dose. The mean theophylline half life before rifampin treatment of 5.11 +/- 0.71 hrs was not different (p greater than 0.10) from the half life of 4.54 +/- 0.71 hrs after treatment with rifampin. In the non-rifampin treated rabbits, the initial mean theophylline half life of 4.78 +/- 1.05 was not different p greater than 0.80) from the half life of 4.94 +/- 1.92 hrs after fourteen days. No significant alterations in clearance or area under the curve were noted for either group. Power analysis indicated that three rabbits were sufficient to detect at least a 25% change in the parameters as being significant (p less than 0.05).


Mutational biosynthesis to generate novel analogs of nosiheptide featuring a fluorinated indolic acid moiety.
Nosiheptide (NOS) is a member of bicyclic thiopeptides possessing a biologically important indolic acid (IA) moiety appended onto the family-characteristic core system. The IA formation relies primarily on NosL, a radical S-adenosylmethionine (SAM) protein that catalyzes a complex rearrangement of the carbon side chain of l-tryptophan, leading to the generation of 3-methyl-2-indolic acid (MIA). Here, we establish an efficient mutational biosynthesis strategy for the structural expansion of the side-ring system of NOS. The nosL-deficient mutant Streptomyces actuosus SL4005 complemented by chemically feeding 6-fluoro-MIA is capable of accumulating two new products. The target product 6'-fluoro-NOS contains an additional fluorine atom at C6 of the IA moiety, in contrast with an unexpected product 6'-fluoro-NOSint that features an open side ring and a bis-dehydroalanine (Dha) tail. The newly obtained 6'-fluoro-NOS displayed equivalent or slightly reduced activities against the tested drug-resistant pathogens compared with NOS, but dramatically decreased water solubility compared with NOS. Our results indicate that the modification of the IA moiety of NOS not only affects its biological activity but also affects its activity which will be key considerations for further modification.


A New Measure for Neural Compensation Is Positively Correlated With Working Memory and Gait Speed.
Neuroimaging studies suggest that older adults may compensate for declines in brain function and cognition through reorganization of neural resources. A limitation of prior research is reliance on between-group comparisons of neural activation (e.g., younger vs. older), which cannot be used to assess compensatory ability quantitatively. It is also unclear about the relationship between compensatory ability with cognitive function or how other factors such as physical exercise modulates compensatory ability. Here, we proposed a data-driven method to semi-quantitatively measure neural compensation under a challenging cognitive task, and we then explored connections between neural compensation to cognitive engagement and cognitive reserve (CR). Functional and structural magnetic resonance imaging scans were acquired for 26 healthy older adults during a face-name memory task. Spatial independent component analysis (ICA) identified visual, attentional and left executive as core networks. Results show that the smaller the volumes of the gray matter (GM) structures within core networks, the more networks were needed to conduct the task (r = -0.408, p = 0.035). Therefore, the number of task-activated networks controlling for the GM volume within core networks was defined as a measure of neural compensatory ability. We found that compensatory ability correlated with working memory performance (r = 0.528, p = 0.035). Among subjects with good memory task performance, those with higher CR used fewer networks than subjects with lower CR. Among poor-performance subjects, those using more networks had higher CR. Our results indicated that using a high cognitive-demanding task to measure the number of activated neural networks could be a useful and sensitive measure of neural compensation in older adults.


Use of drains in foot surgery.
The purpose of this article is to describe how drains can be effective for foot surgeons. Emphasis is placed upon the closed-wound drainage system which was designed for outpatient surgery. Removing transudative or exudative fluids will promote the healing time of a wound that would otherwise delay tissue granulation. These drains eliminate dead space by exerting a negative pressure by means of a perforated catheter, tube, and reservoir. Edema is decreased, and layers of tissue are allowed to adhere faster. The portable closed wound drainage system removes drainage against gravity to allow faster rehabilitation.


Influence of sexual risk behaviour and STI co-infection dynamics on the evolution of HIV set point viral load in MSM.
HIV viral load (VL) is an important predictor of HIV progression and transmission. Anti-retroviral therapy (ART) has been reported to reduce HIV transmission by lowering VL. However, apart from this beneficial effect, increased levels of population mean set-point viral load (SPVL), an estimator for HIV virulence, have been observed in men who have sex with men (MSM) in the decade following the introduction of ART in The Netherlands. Several studies have been devoted to explain these counter-intuitive trends in SPVL. However, to our knowledge, none of these studies has investigated an explanation in which it arises as the result of a sexually transmitted infection (STI) co-factor in detail. In this study, we adapted an event-based, individual-based model to investigate how STI co-infection and sexual risk behaviour affect the evolution of HIV SPVL in MSM before and after the introduction of ART. The results suggest that sexual risk behaviour has an effect on SPVL and indicate that more data are needed to test the effect of STI co-factors on SPVL. Furthermore, the observed trends in SPVL cannot be explained by sexual risk behaviour and STI co-factors only. We recommend to develop mathematical models including also factors related to viral evolution as reported earlier in the literature. However, this requires more complex models, and the collection of more data for parameter estimation than what is currently available.


Medicare recoupment: a legal and legislative update.
In 1987, the Texas Medical Association and 20 other plaintiffs filed suit against the US Department of Health and Human Services (HHS) over the federal government's effort to recoup $14 million from physicians and patients for alleged overpayments during July 1985 through March 31, 1986. A federal district court judge granted a preliminary injunction that halted the recoupment pending a full hearing on the merits of the case and a decision. In June 1989 the Court of Appeals overturned the district court's decision, and TMA turned to the US Congress for relief. This article summarizes recent judicial and legislative progress in halting the recoupment.