Identification of the critical region of 12p over-representation in testicular germ cell tumors of adolescents and adults.
Cytogenetically, testicular germ cell tumors of adolescents and adults (TGCTs) are characterized by gain of 12p-sequences, most often through isochromosome formation (i(12p)). Fluorescence in situ hybridization (FISH) has shown that i(12p))-negative TGCTs also cryptically contain extra 12p-sequences. The consistency of 12p-over-representation in all histological subtypes of TGCTs, including their preinvasive stage, suggests that gain of one or more genes on 12p is crucial in the development of this cancer. So far, studies aimed at the identification of the relevant gene(s) were based on the 'candidate-gene approach'. No convincing evidence in favor of or against a particular gene has been reported. We combined conventional karyotyping, comparative genomic hybridization, and FISH to identify TGCTs with amplifications of restricted regions of 12p. Out of 49 primary TGCTs (23 without i(12p), 13 with and 13 unknown), eight tumors (six without i(12p) and two unknown) showed amplifications corresponding to 12p11.1-p12.1. Using bicolour-FISH, physical mapping, and semi-quantitative polymerase chain reactions, the size of the shortest region of overlap of amplification (SROA) was estimated to be between 1750-3000 kb. In addition, we mapped a number of genes in and around this region. While fourteen known genes could be excluded as candidates based on their location outside this region, we demonstrate that KRAS2, JAW1 and SOX5 genes are localized within the SROA. While KRAS2 and JAW1 map to the proximal border of the SROA, SOX5 maps centrally in the SROA. KRAS2 and JAW1 are expressed in all TGCTs, whereas one 12p amplicon-positive TGCT lacks expression of SOX5. The critical region of 12p over-represented in TGCTs is less than 8% of the total length of the short arm of chromosome 12. It will be helpful in the identification of the gene(s) involved in TGCT-development.


Mechanisms of noncovalent β subunit regulation of NaV channel gating.
Voltage-gated Na+ (NaV) channels comprise a macromolecular complex whose components tailor channel function. Key components are the non-covalently bound β1 and β3 subunits that regulate channel gating, expression, and pharmacology. Here, we probe the molecular basis of this regulation by applying voltage clamp fluorometry to measure how the β subunits affect the conformational dynamics of the cardiac NaV channel (NaV1.5) voltage-sensing domains (VSDs). The pore-forming NaV1.5 α subunit contains four domains (DI-DIV), each with a VSD. Our results show that β1 regulates NaV1.5 by modulating the DIV-VSD, whereas β3 alters channel kinetics mainly through DIII-VSD interaction. Introduction of a quenching tryptophan into the extracellular region of the β3 transmembrane segment inverted the DIII-VSD fluorescence. Additionally, a fluorophore tethered to β3 at the same position produced voltage-dependent fluorescence dynamics strongly resembling those of the DIII-VSD. Together, these results provide compelling evidence that β3 binds proximally to the DIII-VSD. Molecular-level differences in β1 and β3 interaction with the α subunit lead to distinct activation and inactivation recovery kinetics, significantly affecting NaV channel regulation of cell excitability.


Attempts to transmit porcine reproductive and respiratory syndrome virus by aerosols under controlled field conditions.
An experimental infection with porcine reproductive and respiratory syndrome virus (PRRSV) was established in 150 five-month-old pigs housed in a fan-ventilated finishing facility, the infected barn. To determine whether air exhausted from the wall fans contained infectious PRRSV, a trailer containing 10 four-week-old PRRSV-naive sentinel pigs was placed 10 m from the building from day 3 after the 150 pigs were infected until day 10. To connect the two airspaces, one end of an opaque plastic tube, 15 m in length and 5 cm in diameter, was fastened to the wall fan of the infected barn, and the other end was placed inside the trailer. Air from the building was exhausted into the trailer 24 hours a day for seven consecutive days and PRRSV infection was monitored in the infected pigs and the sentinel pigs. Air samples were collected from the infected barn and the trailer. PRRSV infection was detected in the infected pigs three and seven days after they were infected, but not in the sentinel pigs. All the air samples were negative for PRRSV by PCR, virus isolation and a pig bioassay.


The MKK(3/6)-p38-signaling cascade alters the subcellular distribution of hnRNP A1 and modulates alternative splicing regulation.
Individual members of the serine-arginine (SR) and heterogeneous nuclear ribonucleoprotein (hnRNP) A/B families of proteins have antagonistic effects in regulating alternative splicing. Although hnRNP A1 accumulates predominantly in the nucleus, it shuttles continuously between the nucleus and the cytoplasm. Some but not all SR proteins also undergo nucleo-cytoplasmic shuttling, which is affected by phosphorylation of their serine/arginine (RS)-rich domain. The signaling mechanisms that control the subcellular localization of these proteins are unknown. We show that exposure of NIH-3T3 and SV-40 transformed green monkey kidney (COS) cells to stress stimuli such as osmotic shock or UVC irradiation, but not to mitogenic activators such as PDGF or EGF, results in a marked cytoplasmic accumulation of hnRNP A1, concomitant with an increase in its phosphorylation. These effects are mediated by the MKK(3/6)-p38 pathway, and moreover, p38 activation is necessary and sufficient for the induction of hnRNP A1 cytoplasmic accumulation. The stress-induced increase in the cytoplasmic levels of hnRNP A/B proteins and the concomitant decrease in their nuclear abundance are paralleled by changes in the alternative splicing pattern of an adenovirus E1A pre-mRNA splicing reporter. These results suggest the intriguing possibility that signaling mechanisms regulate pre-mRNA splicing in vivo by influencing the subcellular distribution of splicing factors.


Isotope-dilution analysis of rate-limiting steps and pools affecting the incorporation of thymidine and deoxycytidine into cultured thymus cells.
1. Rat thymus cells were incubated in homologous serum (10%) and medium 199. The effects of various quantities of thymidine or deoxycytidine (0-30mum) on the radioactive labelling of cells with the corresponding radioactive deoxynucleoside were examined. From plots of the reciprocal of the radioactivity incorporated against the total deoxynucleoside concentration (;isotope-dilution plots'), values were obtained for (a) the V(max.) of the rate-limiting step governing incorporation of the deoxynucleoside, and (b) the concentration of the pool of compounds competing with the radioactive deoxynucleoside at that rate-limiting step. From changes in these values under different experimental conditions inferences were drawn on the position and control of the rate-limiting step within intact cells. 2. Isotope-dilution plots for deoxycytidine were linear, whereas plots for thymidine were bimodal, indicating an abrupt increase in both the V(max.) and pool concentration at a critical thymidine concentration (approx. 5mum). The bimodality was removed by amethopterin. The V(max.) determined with deoxy[U-(14)C]cytidine was approximately equal to the sum of the V(max.) determined with deoxy[5-(3)H]cytidine and the V(max.) determined with [Me-(3)H]thymidine at thymidine concentrations above 5mum. 3. The thymidine competitor pool at thymidine concentrations above 5mum was approximately equal to the sum of the deoxycytidine competitor pool and the thymidine competitor pool at thymidine concentrations below 5mum. The pools were independent of cell concentration and dependent on serum concentration. 4. These results were explained on the following basis. Deoxycytidine in serum (16mum) is the major source of both cytosine and, by way of thymidylate synthetase, thymine, in the DNA of thymus cells. Serum deoxycytidine normally maintains a sufficient intracellular concentration of dTTP to inhibit partially the activity of thymidine kinase. When the dTTP concentration is lowered, either by decreasing the concentration of deoxycytidine or by inhibiting thymidylate synthetase, the activity of thymidine kinase increases. The activity of thymidine kinase may also be increased by concentrations of thymidine greater than 5mum, which overcome the inhibition of the enzyme by dTTP. At concentrations of thymidine below 5mum, thymidine kinase limits the rate of labelling with [Me-(3)H]thymidine and the radioactivity is diluted by a pool of unlabelled thymidine in serum (4mum). At thymidine concentrations greater than 5mum, the activity of DNA polymerase limits the rate of labelling and the radioactivity is diluted both by serum thymidine and, indirectly, by serum deoxycytidine.


Biophysical and X-ray crystallographic analysis of Mps1 kinase inhibitor complexes.
The dual-specificity protein kinase monopolar spindle 1 (Mps1) is a central component of the mitotic spindle assembly checkpoint (SAC), a sensing mechanism that prevents anaphase until all chromosomes are bioriented on the metaphase plate. Partial depletion of Mps1 protein levels sensitizes transformed, but not untransformed, human cells to therapeutic doses of the anticancer agent Taxol, making it an attractive novel therapeutic cancer target. We have previously determined the X-ray structure of the catalytic domain of human Mps1 in complex with the anthrapyrazolone kinase inhibitor SP600125. In order to validate distinct inhibitors that target this enzyme and improve our understanding of nucleotide binding site architecture, we now report a biophysical and structural evaluation of the Mps1 catalytic domain in the presence of ATP and the aspecific model kinase inhibitor staurosporine. Collective in silico, enzymatic, and fluorescent screens also identified several new lead quinazoline Mps1 inhibitors, including a low-affinity compound termed Compound 4 (Cpd 4), whose interaction with the Mps1 kinase domain was further characterized by X-ray crystallography. A novel biophysical analysis demonstrated that the intrinsic fluorescence of SP600125 changed markedly upon Mps1 binding, allowing spectrophotometric displacement analysis and determination of dissociation constants for ATP-competitive Mps1 inhibitors. By illuminating the structure of the Mps1 ATP-binding site our results provide novel biophysical insights into Mps1-ligand interactions that will be useful for the development of specific Mps1 inhibitors, including those employing a therapeutically validated quinazoline template.


Feasibility and Acceptability of a Codesigned Health Care Transition Intervention for Young People With Spinal Cord Injuries.
Due in part to medical complications, adults with a pediatric onset spinal cord injury (SCI) are at higher risk of experiencing dissatisfaction with life and lower perceived physical health when compared to their peers with no disability. To support the prevention of medical complications, young people with SCI must successfully transition to adult health care. Health care transition (HCT) interventions can support young people with chronic conditions in their move to adult health care. To evaluate the feasibility and acceptability of a web-based HCT intervention codesigned with young people with SCI and parents/caregivers. Semi-structured individual interviews were conducted online with young people with SCI and parents/caregivers who transitioned or were preparing for the transition from pediatric to adult health care. Interviews were also conducted with health care professionals. The interviews were analyzed using a hybrid deductive and inductive qualitative content analysis process. Feasibility and acceptability were measured using Bowen and colleagues' framework, which includes eight focus areas: acceptability, demand, implementation, practicality, adaption, integration, expansion, and limited efficacy. Overall, participants responded positively to the intervention and believed that it would be useful to young people with SCI and parents/caregivers. Two areas of Bowen et al.'s framework, implementation and integration, require further consideration in terms of how to embed the intervention into the current transition process. This study found the HCT intervention to be an innovative approach to support young people with SCI and their parent/caregivers that demonstrates promise in the areas of feasibility and acceptability.


Clinical uses of cervical vestibular-evoked myogenic potential testing in pediatric patients.
To demonstrate the feasibility and clinical significance of cervical vestibular-evoked myogenic potential (cVEMP) test in pediatric patients.Retrospective review study was conducted in a pediatric tertiary care facility. A total of 278 patients were identified with adequate data, including medical notes, results of cVEMP, and imaging studies.Among the total of 278 pediatric patients, only 3 children were not able to finish the cVEMP test successfully. In about 90% of the cases, the cVEMP test was requested to investigate a patient's hearing loss and/or vestibular complaints. Over 90% of the cVEMP tests were ordered by specialists such as pediatric otolaryngologists or otologists. Obtained cVEMP results provided useful information in clinical diagnosis and management in all cases.It is feasible to conduct cVEMP testing in children, including infants, and cVEMP testing can provide valuable information in the diagnosis and management of hearing loss and vestibular impairment. This simple and noninvasive test should be embraced by pediatric professionals.


Existing evidence summarization methods cannot guarantee trustworthy patient decision aids.
Our aim was to evaluate how organizations that develop patient decision aids conduct their evidence summarization process and assess whether their current processes provide sufficient information to instill confidence that patient decision aids are trustworthy and up to date. We identified 23 organizations from a public inventory of patient decision aid developers and included only organizations that have produced five or more tools. These organizations were asked to complete a 17-item survey and to share relevant documents. Of the 23 organizations, 18 completed the survey, and 15 were eligible for analysis. Most organizations reported using existing systematic reviews and clinical practice guidelines. Seven of 15 had a documented approach for summarizing evidence, but the documents offered varying levels of detail. Common steps identified are tool-relevant question formation, search strategies, evidence appraisals, and updating policies. Organizations do not use a standardized process to summarize evidence for the patient decision aids that they develop. This is problematic, given that the information they contain is known to influence patients' decisions. Further attention to how organizations summarize evidence for these tools is required.


Prolonged increases in neurotrophic activity associated with kainate-induced hippocampal synaptic reorganization.
Synaptic reorganization occurs in the hippocampus following various forms of seizure activity and injury, and may contribute to epileptogenesis. To address the hypothesis that neurotrophic factors play an inductive role in synaptic reorganization following seizures, we directly measured neurotrophic activity in rat hippocampal extracts after kainate injection or prolonged stimulation of the perforant path. Serial dilutions of hippocampal extracts were added to cultures of chick dorsal root ganglia, which are known to require trophic support from nerve growth factor and other neurotrophins, or ciliary ganglia neurons, which require trophic support from ciliary neurotrophic factor. Neurotrophic activity was significantly increased in hippocampal extracts harvested from 12 h to 2 months after kainate treatment, with the peak effect seen at seven days. This neurotrophic activity was substantially blocked by an anti-nerve growth factor antibody. Extracts at seven days also showed a significant increase in ciliary neurotrophic factor-like activity. Sulfide/silver histochemistry, which stains dentate granule cell axon terminals, revealed that mossy fiber sprouting was evident two weeks following kainate treatment and increased progressively over the next two to six weeks. Perforant path stimulation that produced hyperexcitability in the dentate gyrus, but no sprouting, failed to induce changes in neurotrophic activity. These results suggest there are significant increases in neurotrophic factors following kainate-induced seizures, and the increases may be related to kainate-induced hippocampal injury rather than seizures per se. Furthermore, the timecourse of increased neurotrophic activity parallels that of mossy fiber reorganization, and is consistent with the hypothesis that neurotrophic factors play a role in the injury-induced synaptic reorganization seen in epilepsy.


The effect of Angiojet rheolytic thrombectomy in the endovascular treatment of lower extremity deep venous thrombosis.
Objective To evaluate the clinical safety and effectiveness of percutaneous rheolytic thrombectomy in patients with acute lower extremity deep venous thrombosis. Method Sixty-eight consecutive patients with acute massive lower extremity deep venous thrombosis were included in this retrospective study. A percutaneous rheolytic thrombectomy device (Angiojet ® Rheolytic thrombectomy catheter, Boston Scientific, Marlborough, MA, USA) was used in all patients in an angiography suite through ipsilateral popliteal vein access. Thrombus clearance and complications were evaluated. Furthermore, patients underwent a clinical evaluation according to a modified Villalta scale for the investigation of post thrombotic syndrome in follow-up. The Venous Clinical Severity Score, Venous Insufficiency Epidemiological and Economic Study-Quality of Life/Sym questionnaires were completed preoperatively and readministered postoperatively. Results Overall thrombus clearance (complete recanalization was achieved in 58 patients (85.2%) and partial recanalization was achieved in 7 patients (10.2%) confirmed through venographic assessment was achieved in 95.5% of the patient population. The mean Venous Clinical Severity Score preoperatively was 13.1 ± 2.2 and decreased to 4.0 ± 1.3 postoperatively (P < 0.01). The Villalta scale dropped from 12.9 ± 2.8 to 5.5 ± 1.4 postoperatively (P < 0.001). Overall quality of life and symptoms improved as assessed by Venous Insufficiency Epidemiological and Economic Study-Quality of Life/Sym (P < 0.01 and 0.02, respectively). Only three minor bleedings were seen but none of the patients suffered from major bleeding, symptomatic pulmonary embolism, death, or other procedure related complications. Fifty-nine out of 65 patients (90.7%) who were treated successfully with rheolytic thrombectomy remained patent at 12 months according to DUS and five patients (7.3%) developed a mild post thrombotic syndrome. Conclusion Rheolytic thrombectomy with or without stenting is superior to anticoagulant therapy alone in terms of both ensuring venous patency and improving clinical symptoms. This technique is a safe, effective and easily performed method of endovascular treatment with a low rate of major treatment complications and shows promising clinical mid-term results.


Managing curricular change in the UWI medical schools.
The ideal operational curriculum is dynamic. It is alive, constantly responding to changes within the social milieu served by its programmes. The medical curriculum of the University of the West Indies (UWI) has not been readily responsive to its catchment society's changing needs. This lack of resilience has created both curricular and administrative problems that have remained unsolved. Now, at the threshold of the twenty-first century, many more fundamental curricular changes are imperative in the UWI medical programme if the Faculty of Medical Sciences (FMS) is to be able to withstand the territorial invasions imminent from the global digital institutions of the new age. The new changes that will place the medical curriculum in line with the demands of the new Information Age will also remove the obnoxious effects of the 'dual curriculum' question and related issues. The Change Formula (Ch = V x P x D > C) that has worked the corporate transformations and realignments of the late twentieth century is applied to the thoughts of a reformed management of the UWI medical curriculum, and its ability to break down walls of resistance to change and liberate the curriculum to full dynamism is discussed.


Predictors of host specificity among behavior-manipulating parasites.
A trade-off between resource-specialization and the breadth of the ecological niche is one of the most fundamental biological characteristics. A true generalist (Jack-of-all-trades) displays a broad ecological niche with little resource specialization while the opposite is true for a resource-specialist that has a restricted ecological niche that it masters. Parasites that manipulate hosts' behavior are often thought to represent resource-specialists based on a few spectacular examples of manipulation of the host's behavior. However, the determinants of which, and how many, hosts a manipulating parasite can exploit (i.e., niche breadth) are basically unknown. Here, I present an analysis based on published records of the use of hosts by 67 species from 38 genera of helminths inducing parasite increased trophic transmission, a widespread strategy of parasites that has been reported from many taxa of parasites and hosts. Using individual and multivariate analyses, I examined the effect of the host's and parasite's taxonomy, location of the parasite in the host, type of behavioral change, and the effect of debilitation on host-specificity, measured as the mean taxonomic relatedness of hosts that a parasite can manipulate. Host-specificity varied substantially across taxa suggesting great variation in the level of resource-specialization among manipulating parasites. Location of the parasite, level of debilitation, and type of host were all significant predictors of host-specificity. More specifically, hosts' behavioral modification that involves interaction with the central nervous system presumably restricts parasites to more closely related hosts than does manipulation of the host's behavior via debilitation of the host's physiology. The results of the analysis suggest that phylogenetic relatedness of hosts is a useful measure of host-specificity in comparative studies of the complexity of interactions taking place between manipulating parasites and their hosts.


Associations of active commuting with body fat and visceral adipose tissue: A cross-sectional population based study in the UK.
The promotion of active travel (walking and cycling) is one promising approach to prevent the development of obesity and related cardio-metabolic disease. However the associations between active travel and adiposity remain uncertain. We used the Fenland study (a population based-cohort study; Cambridgeshire, UK, 2005-15) to describe the association of commuting means with DEXA measured body fat and visceral adipose tissue (VAT) among commuters (aged 29-65years; n=7680). We stratified our sample into those living near (within five miles) and far (five miles or further) from work, and categorised commuting means differently for each group reflecting their different travel options. Associations were adjusted for age, education, Mediterranean diet score, smoking, alcohol consumption, test site and either self-reported physical activity or objective physical activity. Among those living near to work, people who reported regularly cycling to work had lower body fat than those who only used the car (adjusting for self-reported physical activity: women, -1.74%, 95% CI: -2.27% to -0.76%; men, -1.30%, -2.26% to -0.33%). Among those who lived far from work, people who reported regular car-use with active travel had lower body fat (women; -1.18%, 95% CI: -2.23% to -0.13%; men, -1.19%, -1.93% to -0.44%). Findings were similar for VAT and when adjusting for objectively measured physical activity instead of self-reported physical activity. In conclusion, active commuting may reduce adiposity and help prevent related cardio-metabolic disease. If people live too far from work to walk or cycle the whole journey, incorporating some active travel within the commute is also beneficial.


Cytogenetics and genomics in pediatric acute lymphoblastic leukaemia.
The last five decades have witnessed significant improvement in diagnostics, treatment and management of children with acute lymphoblastic leukaemia (ALL). These advancements have become possible through progress in our understanding of the genetic and biological background of ALL, resulting in the introduction of risk-adapted treatment and novel therapeutic targets, e.g., tyrosine kinase inhibitors for BCR::ABL1-positive ALL. Further advances in the taxonomy of ALL and the discovery of new genetic biomarkers and therapeutic targets, as well as the introduction of targeted and immunotherapies into the frontline treatment protocols, may improve management and outcome of children with ALL. In this review we describe the current developments in the (cyto)genetic diagnostics and management of children with ALL, and provide an overview of the most important advances in the genetic classification of ALL. Furthermore, we discuss perspectives resulting from the development of new techniques, including artificial intelligence (AI).


Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida.
Invasive lionfish (Pterois volitans/miles) pose a serious threat to marine ecosystems throughout the western Atlantic Ocean and Caribbean Sea. The development of a fishery for lionfish has been proposed as a strategy for controlling populations; however, there is concern about consumption of this species by humans due to its high trophic position and potential for bioaccumulation of mercury. We analyzed total mercury (THg) in tissues of lionfish from two locations on the east coast of Florida. THg in lionfish increased with size and differed by location and sex. THg was highest in muscle tissue and was strongly positively correlated among tissues. THg in lionfish was lower than other commonly consumed marine fishes, and falls into Florida's least restrictive advisory level. Consumption of lionfish poses a low risk and concerns over mercury bioaccumulation should not present a significant barrier to lionfish harvest.


Cortical plasticity in patients with median nerve lesions studied with MEG.
We have previously shown age- and time-dependent effects on brain activity in the primary somatosensory cortex (SI), in a functional magnetic resonance imaging (fMRI) study of patients with median nerve injury. Whereas fMRI measures the hemodynamic changes in response to increased neural activity, magnetoencephalography (MEG) offers a more concise way of examining the evoked response, with superior temporal resolution. We therefore wanted to combine these imaging techniques to gain additional knowledge of the plasticity processes in response to median nerve injury. Nine patients with median nerve trauma at the wrist were examined with MEG. The N1 and P1 responses at stimulation of the injured median nerve at the wrist were lower in amplitude compared to the healthy side (p < .04). Ulnar nerve stimulation of the injured hand resulted in larger N1 amplitude (p < .04). The amplitude and latency of the response did not correlate with the sensory discrimination ability. There was no correlation between N1 amplitude and size of cortical activation in fMRI. There was no significant difference in N1 latency between the injured and healthy median nerve. N1 latency correlated positively with age in both the median and ulnar nerve, and in both the injured and the healthy hand (p < .02 or p < .001). It is concluded that conduction failure in the injured segment of the median nerve decreases the amplitude of the MEG response. Disinhibition of neighboring cortical areas may explain the increased MEG response amplitude to ulnar nerve stimulation. This can be interpreted as a sign of brain plasticity.


High-Dose Compared With Standard-Dose Oxytocin Regimens to Augment Labor in Nulliparous Women: A Randomized Controlled Trial.
To evaluate whether a high-dose oxytocin regimen reduces the risk for primary cesarean birth and other obstetric morbidities when compared with standard dosing. In a double-blind randomized clinical trial of nulliparous women at or beyond 36 weeks of gestation who were undergoing augmentation of labor, participants were assigned to high-dose (initial and incremental rates of 6 milliunits/min) or standard-dose (initial and incremental rates of 2 milliunits/min) oxytocin regimens. The primary outcome was cesarean birth. Prespecified secondary outcomes included labor duration, clinical chorioamnionitis, endometritis, postpartum hemorrhage, Apgar score 3 or less at 5 minutes, umbilical artery acidemia, neonatal intensive care unit admission, perinatal death, and a severe perinatal morbidity composite. A sample size of 501 per group (n=1,002) was planned to detect a 6.6% absolute reduction in rate of the primary outcome, from 20% in the standard-dose group to 13.4% in the high-dose group with 80% power. From September 2015 to September 2020, 1,003 participants were randomized-502 assigned to high-dose and 501 assigned to standard dosing. The majority of participants were of White race, were married or living as married, and had commercial insurance. Baseline characteristics between groups were similar. The primary outcome occurred in 14.5% of those receiving high-dose compared with 14.4% of those receiving standard-dose oxytocin (relative risk, 1.01; 95% CI 0.75-1.37). The high-dose group had a significantly shorter mean labor duration (9.1 vs 10.5 hours; P<.001), and a significantly lower chorioamnionitis incidence (10.4% vs 15.6%; relative risk, 0.67; 95% CI 0.48-0.92) compared with standard dosing. Umbilical artery acidemia was significantly less frequent in the high-dose group in complete case analysis, but this finding did not persist after multiple imputation (relative risk, 0.55; 95% CI 0.29-1.04). There were no significant differences in other secondary outcomes. Among nulliparous participants who were undergoing augmentation of labor, a high-dose oxytocin regimen, compared with standard dosing, did not affect the cesarean birth risk but significantly reduced labor duration and clinical chorioamnionitis frequency without adverse effects on perinatal outcomes. ClinicalTrials.gov, NCT02487797.


Towards Biohybrid Lung: Induced Pluripotent Stem Cell Derived Endothelial Cells as Clinically Relevant Cell Source for Biologization.
In order to provide an alternative treatment option to lung transplantation for patients with end-stage lung disease, we aim for the development of an implantable biohybrid lung (BHL), based on hollow fiber membrane (HFM) technology used in extracorporeal membrane oxygenators. Complete hemocompatibility of all blood contacting surfaces is crucial for long-lasting BHL durability and can be achieved by their endothelialization. Autologous endothelial cells (ECs) would be the ideal cell source, but their limited proliferation potential excludes them for this purpose. As induced pluripotent stem cell-derived ECs enable the generation of a large number of ECs, we assessed and compared their capacity to form a viable and confluent monolayer on HFM, while indicating physiologic EC-specific anti-thrombogenic and anti-inflammatory properties. ECs were generated from three different human iPSC lines, and seeded onto fibronectin-coated poly-4-methyl-1-pentene (PMP) HFM. Following phenotypical characterization, ECs were analyzed for their thrombogenic and inflammatory behavior with or without TNFα induction, using FACS and qRT-PCR. Complementary, leukocyte- and platelet adhesion assays were carried out. The capacity of the iPSC-ECs to reendothelialize cell-free monolayer areas was assessed in a scratch assay. ECs sourced from umbilical cord blood (hCBECs) were used as control. iPSC-derived ECs formed confluent monolayers on the HFM and showed the typical EC-phenotype by expression of VE-cadherin and collagen-IV. A low protein and gene expression level of E-selectin and tissue factor was detected for all iPSC-ECs and the hCBECs, while a strong upregulation of these markers was noted upon stimulation with TNFα. This was in line with the physiological and strong induction of leukocyte adhesion detected after treatment with TNFα, iPSC-EC and hCBEC monolayers were capable of reducing thrombocyte adhesion and repopulating scratched areas. iPSCs offer the possibility to provide patient-specific ECs in abundant numbers needed to cover all blood contacting surfaces of the BHL with a viable, non-thrombogenic and non-inflammatory monolayer. iPSC-EC clones can differ in terms of their reendothelialization rate, and pro-inflammatory response. However, a less profound inflammatory response may even be advantageous for BHL application. With the proven ability of the seeded iPSC-ECs to reduce thrombocyte adhesion, we expect that thrombotic events that could lead to BHL occlusion can be avoided, and thus, justifies further studies on enabling BHL long-term application.


Nucleotide sequence of two rasH related-genes isolated from the yeast Saccharomyces cerevisiae.
A complete nucleotide sequence of two ras-related yeast genes (c- rassc -1 and c- rassc -2) isolated from the yeast strain Saccharomyces cerevisiae is reported. They encode predicted polypeptides of 40,000 and 41,000 daltons, respectively. The N-terminal 170 amino acids from both genes show extensive amino acid homology to other ras genes from vertebrates, whereas their C-termini have diverged. These genes should be useful in the elucidation of a normal biological function of ras-related genes in a simple system like yeast.


Clinical Features of Heart Failure With Normal Ejection Fraction: Insights From the ASIAN-HF Registry.
Heart failure and left ventricular ejection fraction in the normal range (HFnEF) (left ventricular ejection fraction [LVEF] of ≥55% for men and ≥60% for women) is understudied. The authors aimed to characterize patients with HFnEF compared with those with preserved (≥50%) yet below the normal LVEF. In an Asian HF registry, clinical characteristics, echocardiographic features, and outcomes were compared across: 1) HFnEF; 2) heart failure with preserved left ventricular ejection fraction (HFpEF) (LVEF of ≥50%) and below normal LVEF; and 3) community-based controls without HF. Cluster analysis of echocardiographic parameters was performed and validated in an external cohort. Among 1,765 patients with HFpEF (age 68 ± 12 years; 50% women), 1,313 (74.4%) had HFnEF. Compared with patients with HFpEF and below normal LVEF, patients with HFnEF had less coronary artery disease (33.7% vs 27.9%), greater LV wall thickness, and higher stroke volume, but similar 2-year age-adjusted all-cause mortality (HR: 0.8; 95% CI: 0.6-1.2). Five echocardiographic clusters with similar 2-year mortality were identified: 1) normal LV (normal structure despite increased filling pressure; least comorbidities) in 25%; 2) restrictive (smallest stroke volume; predominantly elderly women) in 26%; 3) hypertrophic (most concentric hypertrophy; more men) in 25%; 4) high output (greatest stroke volume; predominantly obese younger men) in 10%; and 5) atrial dominant (most left atrial myopathy; mainly elderly women with multiple comorbidities) in 10%. Similar patterns were found in the validation cohort. The majority of patients with HFpEF had normal LVEF, which consists of patients with different patterns of cardiac features and clinical characteristics. Results may carry implications for targeted treatment approaches in HFpEF.


A Qualitative Study on the Practice of Yoga for Women with Pain-Associated Endometriosis.
To understand the meaning women with pain-associated endometriosis attribute to yoga practice regarding their physical and emotional state at the beginning of the practice; pain management by integrating body and mind; secondary benefits of the practice of yoga, such as self-knowledge, self-care, and autonomy; and the role of the yoga group as psychosocial support. Qualitative study conducted simultaneously with a randomized clinical trial. Public university hospital in southeastern Brazil between August 2013 and December 2014. Fifteen women with pain-associated endometriosis who practiced yoga for 8 weeks. After completing the twice-a-week program, all women participated in a single, semi-structured interview. Interviews were recorded and transcribed verbatim, and thematic analyses were performed. The main themes of analysis were women's expectations regarding the practice of yoga, physical and emotional state of women at the beginning of yoga practice, control and pain management through the integration of body and mind, secondary benefits, acquisition of self-knowledge and autonomy, and the role of yoga group as psychosocial support. All participants reported that yoga was beneficial to control pelvic pain. They related that they were aware of the integration of body and psyche during yoga practice and that this helped in the management of pain. Women said they had identified a relationship between pain management and breathing techniques (pranayama) learned in yoga and that breathing increased their ability to be introspective, which relieved pain. The participants have developed greater self-knowledge, autonomy, and self-care and have reduced the use of pain and psychiatric medications. They created ties among themselves, suggesting that the yoga group allowed psychosocial support. Bodily and psychosocial mechanisms to control pain were identified in women with endometriosis. To reach such control, it is crucial that mind and body integrative techniques are learned.


Whole blood production of monocytic cytokines (IL-1beta, IL-6, TNF-alpha, sIL-6R, IL-1Ra) in haemodialysed patients.
The production of monocytic cytokines by isolated mononuclear cells after stimulation by phytohaemagglutinin (PHA) and lipopolysaccharide (LPS) is generally increased in haemodialysed (HD) patients. We performed whole blood (WB) cultures to evaluate cytokine production by blood cells inside their complex cellular and humoral network. Diluted whole blood from HD patients (collected before dialysis) and controls was cultured alone with PHA (2.5 microg/ml) or LPS (1 and 3 microg/ml). Supernatants were collected after 24 and 48 h of culture, and concentrations of IL-1 beta, IL-6, TNF-alpha, sIL-6R and IL-1Ra were determined by ELISA. The low spontaneous production of IL-1beta, IL-6 and TNF-alpha in both patients and controls was not significantly modified by PHA. The lower dose of LPS (1 microg/ml) induced a significant but lower increase in production of IL-1beta, IL-6 and TNF-alpha in patients than in controls. In contrast, while it did not further increase their production in controls, the higher concentration of LPS (3 microg/ml) still increased their production in patients to the same level than in controls. The plasma concentrations of sIL-6R were higher in patients than in controls. In both groups, the sIL-6R concentration did not vary during the culture period whether the cells were stimulated or not with LPS or PHA. This suggests that the increased plasma levels of sIL-6R were not produced by blood cells. Despite a similar significant LPS and PHA induced production of IL-1Ra, the IL-1Ra/IL-1beta ratio was always higher in patients than in controls. Monocytes from HD patients in WB cultures are hyporesponsive to PHA and LPS for their IL-1beta, TNFalpha and IL-6 production in contrast to isolated monocytes that demonstrate signs of activation. If it reflects the in vivo situation it could partly explain the immune defect in uraemic and haemodialysed patients. Higher sIL-6R/IL-6 and IL-1Ra/IL-1beta ratios could also participate to the complex immune disturbances of HD patients by reducing the biological activity of two cytokines playing a major role in the immune and inflammatory network.


Recent developments in the use of peroxygenases - Exploring their high potential in selective oxyfunctionalisations.
Peroxygenases are an emerging new class of enzymes allowing selective oxyfunctionalisation reactions in a cofactor-independent way different from well-known P450 monooxygenases. Herein, we focused on recent developments from organic synthesis, molecular biotechnology and reaction engineering viewpoints that are devoted to bring these enzymes in industrial applications. This covers natural diversity from different sources, protein engineering strategies for expression, substrate scope, activity and selectivity, stabilisation of enzymes via immobilisation, and the use of peroxygenases in low water media. We believe that peroxygenases have much to offer for selective oxyfunctionalisations and we have much to study to explore the full potential of these versatile biocatalysts in organic synthesis.


Speed and quality goals in procedural skills learning: A randomized experiment.
Purpose: Compare time (speed) and product quality goals in a surgical procedural task.Methods: Secondary school students participating in a medical simulation-based training activity participated in a randomized experiment. Each participant completed eight repetitions of a blood vessel ligation. Once, between repetitions four and five, each participant received a randomly-assigned speed goal or quality goal. Outcomes included time and leak-free ligatures.Results: 80 students participated. The speed-goal group performed 18% faster on the final repetition than the quality-goal group, with adjusted fold change (FC) 0.82 (95% confidence interval [CI], 0.71, 0.94; p = 0.01). Conversely, the speed-goal group had fewer high-quality (leak-free) ligatures (odds ratio [OR] 0.36 [95% CI, 0.22, 0.58; p < 0.001]). For the quality-goal group, leaky ligatures took longer post-intervention than leak-free ligatures (FC 1.09 [95% CI, 1.02, 1.17; p = 0.01]), whereas average times for leaky and leak-free ligatures were similar for the speed-goal group (FC 0.97 [95% CI, 0.91, 1.04; p = 0.38]). For a given performance time, the speed-goal group had more leaks post-intervention than the quality-goal group (OR 3.35 [95% CI, 1.58, 7.10; p = 0.002]).Conclusions: Speed and quality goals promote different learning processes and outcomes among novices. Use of both speed and quality goals may facilitate more effective and efficient learning.


[Elective splenectomy at landspitali university hospital 1993-2004 efficacy and long-term outcome].
To evaluate the long-term outcome of elective splenectomy, with emphasis on the incidence of complications, vaccine immunization and patient´s knowledge about asplenia. Medical reports of all patients, who underwent elective splenectomy during the time period of 1993-2004, were reviewed. Questionnaire was sent to 96% (44/46) patients alive. The average age was 50 (8-83) years. Thirty-five patients were male and 32 were female. Eighty percent responded to the questionnaire. Most of the patients (31) had idiopathic thrombocytopenic purpura (ITP). Complete response was obtained in 60% (18/30) and partial response in 23% (7/30). Five patients had spherocytosis and all of them had complete response. None of the three patients with autoimmune hemolytic anemia had any response to the splenectomy. Patients were vaccinated against pneumococci in 92% of the cases. In 44% of the cases revaccination was done. Only 41% of those who answered experienced that they had got a good education about the consequences of asplenia. Sixteen percent of the patients (10/64) had major postoperative complications. One patient with metastatic cancer and thrombocytopenia died within 30 days of surgery. Five patients had long-term complications. Two had pneumococcal sepsis, one of them was unvaccinated and the other had not been revaccinated. Splenectomy has a good long-term outcome for spherocytosis and ITP patients. The incidence of complications is high. It is possible that better guidelines and better patient´s education can lower the complication rate and improve the outcome.


Terminology and coding systems of drugs--CDrugs.
The average patient contact that results in a drug prescription depends on the organizational environment. Nevertheless, in any country, more than half of patient contacts results in a prescription. On average in Belgium--a country with a free liberal medicine--more than 80% of the GP-patient contacts results in some kind of medicinal product prescription. Linkage with a drug knowledge base is thus a very important, sometimes even essential, requirement in developing a health information system.


Cognitive dysfunction and functional disability in alcohol-dependent adults with or without a comorbid affective disorder.
Little is known about the relationship between cognitive dysfunction and functional disability in alcohol dependence with comorbid affective disorders. We investigated the neuropsychology of alcohol dependence in detoxified adults with and without affective comorbidity and examined the factors associated with prolonged functional disability. From a total of 42 participants (age range = 18-44 years), 12 out of 21 alcohol-dependent participants had a comorbid affective disorder, 12 had an affective disorder only, and 9 were healthy controls. Participants completed a semi-structured clinical interview, questionnaires and comprehensive neuropsychological assessment. Following detoxification (median = 35 days; M = 41.2 days, SD = 17.9), visual learning and memory functioning was worse in alcohol-dependent individuals. Comorbid affective disorders did not appear to exacerbate cognitive dysfunction. Psychiatric comorbidity and current depressive symptoms were predictive of poorer functional disability. Furthermore, learning and memory, and response inhibition, contributed significantly and independently to predicting functional disability over and above clinical and demographic factors. Psychiatric comorbidity does not appear to be associated with more pronounced neuropsychological dysfunction in alcohol dependence. Conversely, both comorbid affective disorders and cognitive factors were critical in determining the functional outcomes of alcohol-dependent adults recently undergoing medically supervised inpatient detoxification.


What are the Economic Costs to Society Attributable to Alcohol Use? A Systematic Review and Modelling Study.
Alcohol-attributable costs to society are captured by cost-of-illness studies, however estimates are often not comparable, e.g. due to the omission of relevant cost components. In this contribution we (1) summarize the societal costs attributable to alcohol use, and (2) estimate the total costs under the assumption that all cost components are considered. A systematic review and meta-analyses were conducted for studies reporting costs from alcohol consumption for the years 2000 and later, using the EMBASE and MEDLINE databases. Cost estimates were converted into 2019 international dollars (Int$) per adult and into percentage of gross domestic product (GDP). For each study, weights were calculated to correct for the exclusion of cost indicators. Of 1708 studies identified, 29 were included, and the mean costs of alcohol use amounted to 817.6 Int$ per adult (95% confidence interval [CI] 601.8-1033.4), equivalent to 1.5% of the GDP (95% CI 1.2-1.7%). Adjusting for omission of cost components, the economic costs of alcohol consumption were estimated to amount to 1306 Int$ per adult (95% CI 873-1738), or 2.6% (95% CI 2.0-3.1%) of the GDP. About one-third of costs (38.8%) were incurred through direct costs, while the majority of costs were due to losses in productivity (61.2%). The identified cost studies were mainly conducted in high-income settings, with high heterogeneity in the employed methodology. Accounting for some methodological variations, our findings demonstrate that alcohol use continues to incur a high level of cost to many societies. PROSPERO #CRD42020139594.


Burden of chronic diseases associated with periodontal diseases: a retrospective cohort study using UK primary care data.
To identify the association between periodontal diseases (gingivitis and periodontitis) and chronic diseases including cardiovascular disease, cardiometabolic disease, autoimmune disease and mental ill health. Retrospective cohort. IQVIA Medical Research Data-UK between 1 January 1995 and 1 January 2019. 64 379 adult patients with a general practitioner recorded diagnosis of periodontal disease (exposed patients) were matched to 251 161 unexposed patients by age, sex, deprivation and registration date. Logistic regression models accounting for covariates of clinical importance were undertaken to estimate the adjusted OR (aOR) of having chronic diseases at baseline in the exposed compared with the unexposed group. Incidence rates for each outcome of interest were then provided followed by the calculation of adjusted HRs using cox regression modelling to describe the risk of outcome development in each group. The average age at cohort entry was 45 years and the median follow-up was 3.4 years. At study entry, the exposed cohort had an increased likelihood of having a diagnosis of cardiovascular disease (aOR 1.43; 95% CI 1.38 to 1.48), cardiometabolic disease (aOR 1.16; 95% CI 1.13 to 1.19), autoimmune disease (aOR 1.33; 95% CI 1.28 to 1.37) and mental ill health (aOR 1.79; 95% CI 1.75 to 1.83) compared with the unexposed group. During the follow-up of individuals without pre-existing outcomes of interest, the exposed group had an increased risk of developing cardiovascular disease (HR 1.18; 95% CI 1.13 to 1.23), cardiometabolic disease (HR 1.07; 95% CI 1.03 to 1.10), autoimmune disease (HR 1.33; 95% CI 1.26 to 1.40) and mental ill health (HR 1.37; 95% CI 1.33 to 1.42) compared with the unexposed group. In this cohort, periodontal diseases appeared to be associated with an increased risk of developing cardiovascular, cardiometabolic, autoimmune diseases and mental ill health. Periodontal diseases are very common; therefore, an increased risk of other chronic diseases represent a substantial public health burden.


Pregnant women infected with pandemic H1N1pdm2009 influenza virus displayed overproduction of peripheral blood CD69+ lymphocytes and increased levels of serum cytokines.
The first pandemic of the 21st century occurred in 2009 and was caused by the H1N1pdm influenza A virus. Severe cases of H1N1pdm infection in adults are characterized by sustained immune activation, whereas pregnant women are prone to more severe forms of influenza, with increased morbi-mortality. During the H1N1pdm09 pandemic, few studies assessed the immune status of infected pregnant women. The objective of this study was to evaluate the behavior of several immune markers in 13 H1N1pdm2009 virus-infected pregnant (PH1N1) women, in comparison to pregnant women with an influenza-like illness (ILI), healthy pregnant women (HP) and healthy non-pregnant women (HW). The blood leukocyte phenotypes and the serological cytokine and chemokine concentrations of the blood leukocytes, as measured by flow cytometry, showed that the CD69+ cell counts in the T and B-lymphocytes were significantly higher in the PH1N1 group. We found that pro-inflammatory (TNF-α, IL-1β, IL-6) and anti-inflammatory (IL-10) cytokines and some chemokines (CXCL8, CXCL10), which are typically at lower levels during pregnancy, were substantially increased in the women in the ILI group. Our findings suggest that CD69 overexpression in blood lymphocytes and elevated levels of serum cytokines might be potential markers for the discrimination of H1N1 disease from other influenza-like illnesses in pregnant women.


FLASH Radiotherapy Using Single-Energy Proton PBS Transmission Beams for Hypofractionation Liver Cancer: Dose and Dose Rate Quantification.
This work aims to study the dose and ultra-high-dose rate characteristics of transmission proton pencil beam scanning (PBS) FLASH radiotherapy (RT) for hypofractionation liver cancer based on the parameters of a commercially available proton system operating under FLASH mode. An in-house treatment planning software (TPS) was developed to perform intensity-modulated proton therapy (IMPT) FLASH-RT planning. Single-energy transmission proton PBS plans of 4.5 Gy × 15 fractions were optimized for seven consecutive hepatocellular carcinoma patients, using 2 and 5 fields combined with 1) the minimum MU/spot chosen between 100 and 400, and minimum spot time (MST) of 2 ms, and 2) the minimum MU/spot of 100, and MST of 0.5 ms, based upon considerations in target uniformities, OAR dose constraints, and OAR FLASH dose rate coverage. Then, the 3D average dose rate distribution was calculated. The dose metrics for the mean dose of Liver-GTV and other major OARs were characterized to evaluate the dose quality for the different combinations of field numbers and minimum spot times compared to that of conventional IMPT plans. Dose rate quality was evaluated using 40 Gy/s volume coverage (V40Gy/s). All plans achieved favorable and comparable target uniformities, and target uniformity improved as the number of fields increased. For OARs, no significant dose differences were observed between plans of different field numbers and the same MST. For plans using shorter MST and the same field numbers, better sparing was generally observed in most OARs and was statistically significant for the chest wall. However, the FLASH dose rate coverage V40Gy/s was increased by 20% for 2-field plans compared to 5-field plans in most OARs with 2-ms MST, which was less evident in the 0.5-ms cases. For 2-field plans, dose metrics and V40Gy/s of select OARs have large variations due to the beam angle selection and variable distances to the targets. The transmission plans generally yielded inferior dosimetric quality to the conventional IMPT plans. This is the first attempt to assess liver FLASH treatment planning and demonstrates that it is challenging for hypofractionation with smaller fractional doses (4.5 Gy/fraction). Using fewer fields can allow higher minimum MU/spot, resulting in higher OAR FLASH dose rate coverages while achieving similar plan quality compared to plans with more fields. Shorter MST can result in better plan quality and comparable or even better FLASH dose rate coverage.


Enhancement of heavy metal tolerance and accumulation efficiency by expressing Arabidopsis ATP sulfurylase gene in alfalfa.
Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.


Cadherin adhesion and mechanotransduction.
Cadherins are the principal adhesion proteins at intercellular junctions and function as the biochemical Velcro that binds cells together. Besides this mechanical function, cadherin complexes are also mechanotransducers that sense changes in tension and trigger adaptive reinforcement of intercellular junctions. The assembly and regulation of cadherin adhesions are central to their mechanical functions, and new evidence is presented for a comprehensive model of cadherin adhesion, which is surprisingly more complex than previously appreciated. Recent findings also shed new light on mechanisms that regulate cadherin junction assembly, adhesion, and mechanotransduction. We further describe recent evidence for cadherin-based mechanotransduction, and the rudiments of the molecular mechanism, which involves α-catenin and vinculin as key elements. Potential roles of a broader cast of possible force-sensitive partners are considered, as well as known and speculative biological consequences of adhesion and force transduction at cadherin-mediated junctions.


Effect of early clopidogrel discontinuation on rehospitalization in acute coronary syndrome: results from two distinct patient populations.
Results of a study of the association between early discontinuation of clopidogrel therapy and rehospitalization rates among patients with acute coronary syndrome (ACS) are reported. In a retrospective observational study, analyses of two nationally representative cohorts of adults hospitalized for acute myocardial infarction (AMI) or coronary stent insertion were conducted to assess risk factors for ACS-related adverse outcomes (rehospitalization for AMI or coronary revascularization) during the 12 months after completion of an initial 28-day course of clopidogrel therapy. Case data were sourced from a commercial insurance claims database, a pharmacy administrative claims database, and a combined dataset that linked hospital discharge and outpatient service data; a time-varying method was used to differentiate adverse events occurring "on" and "off" clopidogrel therapy. One cohort analysis (n = 42,757) indicated that patients who discontinued clopidogrel early (i.e., within 12 months of index discharge) were at significantly increased risk for ACS-related rehospitalization during the 12-month study period (hazard ratio [HR] = 1.11; 95% confidence interval [CI], 1.02-1.20; p < 0.05). In the other cohort analysis (n = 3,171), early clopidogrel discontinuation was associated with an increased risk of rehospitalization or inpatient death (HR = 1.75; 95% CI, 1.59-1.91; p < 0.0001). Observational evidence from analyses of data on two large cohorts of patients with primarily employer-sponsored health insurance suggests that early discontinuation of clopidogrel therapy after hospitalization for AMI or coronary stent insertion is associated with a significant increase in the risk of ACS-related rehospitalization within the 12-month postdischarge period.


The Association of Predicted Resting Energy Expenditure with Risk of Breast Cancer among Postmenopausal Women in the Women's Health Initiative Cohort.
Obesity and obesity-related metabolic disorders, such as diabetes and chronic inflammation, have been positively associated both with postmenopausal breast cancer and with resting energy expenditure (REE). However, there is limited epidemiologic evidence on the associations between REE and risk of postmenopausal breast cancer. We used multivariable Cox proportional hazards models to examine the association between predicted REE (calculated using the Ikeda, Livingston, and Mifflin equations) and risk of postmenopausal breast cancer overall and by subtypes, and by level of body fat) among 137,283 postmenopausal women in the Women's Health Initiative (WHI). All predicted REEs were positively associated with risk of invasive breast cancer [HRq5 vs. q1 = 1.69; 95% confidence interval (CI), 1.57-1.81; HR = 1.69; 95% CI, 1.57-1.82; and HR = 1.68; 95% CI, 1.56-1.80 for Ikeda, Livingston, and Mifflin, respectively]. These positive associations were observed irrespective of the hormone receptor subtype, grade, and stage of the tumors, but were most pronounced for estrogen receptor-positive/progesterone receptor-positive tumors. After additional adjustment for body mass index (BMI), the associations were mostly attenuated and remained statistically significant for most of the outcomes. We also observed an interaction between the predicted REEs and BMI, with the associations being somewhat stronger among normal weight and overweight women than among obese women (Pinteractions < 0.05). Our findings indicate that relatively high REE is associated with increased risk of invasive breast cancer among postmenopausal women (particularly for the obesity-related tumor subtypes), irrespective of the equation used. Further studies using more objective measures of REE are, however, needed to confirm our findings. This study showed that higher resting energy expenditure (REE) was associated with higher postmenopausal breast cancer risk. REE provides energy to support cancer-associated disorders such as obesity and inflammation. Thus, studies on its association with breast cancer can help to improve our understanding of the pathophysiology of breast cancer.


Metastatic progression is associated with dynamic changes in the local microenvironment.
Most cancer-associated deaths result from metastasis. However, it remains unknown whether the size, microenvironment or other features of a metastatic lesion dictate its behaviour or determine the efficacy of chemotherapy in the adjuvant (micrometastatic) setting. Here we delineate the natural history of metastasis in an autochthonous model of pancreatic ductal adenocarcinoma (PDAC), using lineage tracing to examine the evolution of disseminated cancer cells and their associated microenvironment. With increasing size, lesions shift from mesenchymal to epithelial histology, become hypovascular and accumulate a desmoplastic stroma, ultimately recapitulating the primary tumours from which they arose. Moreover, treatment with gemcitabine and nab-paclitaxel significantly reduces the overall number of metastases by inducing cell death in lesions of all sizes, challenging the paradigm that PDAC stroma imposes a critical barrier to drug delivery. These results illuminate the cellular dynamics of metastatic progression and suggest that adjuvant chemotherapy affords a survival benefit by directly targeting micrometastases.


Metarrhizium anisopliae as a cause of sinusitis in immunocompetent hosts.
Metarrhizium anisopliae is a common pathogen of insects and has even been used to control insect populations. It is rarely isolated from human or animal sources, but recently, there have been three reported cases of disease, two in humans and one in a cat. We present our experience with five isolates from human sources, including two that were the apparent causes of two cases of sinusitis in immunocompetent hosts. The first patient was a 36-year-old male with frontal and ethmoid sinusitis, and the second was a 79-year-old female with chronic sinusitis. Both patients underwent surgery, and pathology of the surgical specimens revealed branching hyphae. Cultures grew only Metarrhizium species. Neither patient received antifungal therapy, and both did well postoperatively. The other three isolates were cultured from bronchoalveolar lavage specimens but were not felt to be clinically significant. Antifungal susceptibility testing using the National Committee for Clinical Laboratory Standards macrobroth method revealed that all isolates were resistant to amphotericin B, 5-flucytosine, and fluconazole. Itraconazole and newer azole compounds were more active. Metarrhizium species may cause disease in humans, even those without evidence of immunosuppression, and are apparently highly resistant to amphotericin B in vitro.


Challenges in the veterinary microbiology diagnostic laboratory: a novel Acinetobacter species as presumptive cause for feline unilateral conjunctivitis.
The present study highlights challenges in the veterinary microbiology diagnostic laboratory in the identification of bacteria responsible for infections in veterinary settings, particularly when evidence-based data is lacking. A 1.8-year-old neutered male domestic cat (FIV/FeLV negative) was presented to a veterinary practice in April 2016 with a history of left unilateral mild conjunctivitis that was empirically treated with fusidic acid and chloramphenicol. In January 2017, the same animal was presented with chronic left unilateral conjunctivitis and an eye swab was submitted for microbiological culture and susceptibility testing. Significant growth was not detected in two samples tested. Finally, in February 2017 another eye swab produced a slow growing pure culture identified by VITEK 2 as Neisseria cinerea (94 % confidence). Given the morphology and multidrug resistance profile of the isolate a 16S rRNA PCR was performed for definitive identification. The nucleotide sequence of the PCR amplicon was 99 % homologous to Acinetobacter equi sp. nov. strain 114. Veterinary microbiology diagnostic laboratories play an important role worldwide, not only in preserving animal health and welfare but also in controlling the spread of zoonotic pathogens. The lack of evidence-based information on the ocular microbiome of healthy cats and the complexity of bacterial ecosystems renders the interpretation of results difficult. A further problem for both the laboratory and the clinician is the lack of interpretive criteria for antibiotic susceptibility test results for some types of infections in animals (including those caused by Acinetobacter ) and the complete unavailability of criteria for topical antibiotic preparations.


Retinal angiographic blood flowmetry is reduced in the ocular ischaemic syndrome.
The aim of this study was to examine the use of quantitative fluorescein angiographic analysis as a means of estimating rates of perfusion of the retina in eyes with a circulatory deficit secondary to carotid artery stenosis. The study included 21 eyes with ocular ischaemic syndrome (OIS) and 20 control eyes from subjects with carotid artery stenosis but without signs of ocular ischaemia. Analyses of video fluorescein angiograms extracting time intervals for the time delay between specific phases of the angiogram were performed. Time delay was compared between groups and in relation to degree of carotid artery stenosis and ocular systolic blood pressure. Among the three flow indices of retinal perfusion (arteriovenous passage time 1 (AVP1), arteriovenous passage time 2 (AVP2) and venous filling time (VP)), those including the venous filling phase were significantly prolonged in the OIS group. Furthermore, AVP2 was delayed by 3 sec. in OIS eyes (16.6 sec. versus 13.6 sec. in controls). VP was 2.4 sec. longer in OIS eyes (11.5 sec. versus 9.1 sec.). We found a significant correlation between AVP2 and ocular perfusion pressure, but no correlation between the degree of carotid artery stenosis and any of the flow indices. In a patient population spanning a wide ocular systolic blood pressure range, angiography-based quantitative flowmetry demonstrated a difference between carotid artery stenosis patients with and without OIS and a correlation between flow and ocular perfusion pressure. While angiographic flowmetry proved effective in discriminating between groups of individuals, it can only be used to support the diagnosis of the ocular ischaemic syndrome in patients with extreme flow reduction.


The effect of nonoxynol-9 and chlorhexidine on HIV and sperm in vitro.
The effect of chlorhexidine and nonoxynol-9, either singly or in combination, on the replication and infectivity of HIV and the survival of both lymphocytes (MT2 cells) and human spermatozoa, was studied in vitro. Exposure of MT2 cells to 200 microg/ml nonoxynol-9 or 1 mg/ml chlorhexidine for one minute destroyed their viability. A combination of 60 microg/ml of nonoxynol-9 and chlorhexidine, however, killed MT2 cells under the same conditions. Nonoxynol-9 and chlorhexidine were both spermicidal, 268 microg/ml nonoxynol-9, or 2.063 mg/ml chlorhexidine caused complete immobilization of sperm after one minute. The same effect was achieved by a combination of 200 microg/ml nonoxynol-9 and 1.0 mg/ml chlorhexidine. The effect of chlorhexidine and nonoxynol-9 on the replication of HIV was estimated by the output of p24 (the HIV core protein) and the concentration of virus was determined by titration with MT2 cells. Separately, 300 microg/ml nonoxynol-9 alone completely inactivated HIV, while 1 mg/ml chlorhexidine was 80%-100% effective. Certain combinations of nonoxynol-9 and chlorhexidine were antagonistic in their inactivation of HIV, up to 400 microg/ml chlorhexidine partly neutralized the action of 200-500 microg/ml nonoxynol-9.


Renal handling of aluminum in the rat: clearance and micropuncture studies.
Previous uncertainty regarding glomerular ultrafilterability (UF) of aluminum has limited the definition of renal Al handling. Glomerular micropuncture was therefore performed in hydropenic Munich-Wistar rats infused with AlCl3 to achieve plasma (P) Al levels between 2 and 10 mg/liter. Glomerular fluid, P, and urine Al concentrations were measured by flameless atomic-absorption spectrophotometry. UFA1 was inversely correlated with PA1 [%UFA1 = 10.3 - 8.4 (log PA1), r = -0.90, P less than 0.01]. When this equation was used to calculate the filtered load (FLA1), A1 excretion (UA1V, ng/min) in simultaneously collected samples was found to be a direct function of FLA1 [UA1V = 5.7 + 0.37 (FLA1), r = 0.93, P less than 0.01]. Fractional excretion (FE) of A1 was 39.4 +/- 4.2% in these hydropenic experiments (FENa = 0.3 +/- 0.1%). We next evaluated the tubular handling of A1 (using these UF data) during step-wise extracellular fluid volume expansion with isotonic saline (2.5, 5.0, 7.0, and 7.0% body wt) and during the infusion of increasing doses (2.7, 5.3, 8.0, and 8.0 mg X kg-1 X h-1) of furosemide as urinary losses were quantitatively replaced. The natriuresis produced by volume expansion (FENa = 1.0, 3.0, 8.4, and 7.9%) and furosemide (FENa = 4.2, 6.0, 6.6, and 6.7%) were comparable. At similar FLA1, 7% volume expansion but not furosemide (at any dose) increased UA1V (240 and 95 ng/min, respectively, vs. 116 ng/min in hydropenia) and FEA1 (84.5 and 29.4 vs. 37.4%, respectively). These data indicate that at pharmacological PA1 levels, less than 8.4% of PA1 is ultrafilterable, suggesting extensive plasma protein binding.(ABSTRACT TRUNCATED AT 250 WORDS)


From the "real-world evidence" to prevention strategies: health inequalities in immigrants residing in an Italian Local Health Unit.
to assess the health inequalities of immigrants in the Local Health Unit (LHU) n.1 of the Abruzzo Region with a real-world approach. In particular, 4 main categories of hospitalisation (MDC) linked to health risk behaviours and social determinants were analysed. retrospective study on hospital discharge data (HDD). resident population in the LHU n.1 of the Abruzzo Region between 2010 and 2018. Hospital discharge forms (HDFs) of the hospitals under the administration of the LHU No. 1 of the Abruzzo Region, which includes three geographical areas (Marsica, Peligno-Sangrina, L'Aquila), in the years 2010-2018 were analysed. crude hospitalisation trends for immigrants and Italians; comparison of standardised hospitalisation rates in the main immigrant communities. overall, 26,570 SDO were analysed. Average age of inpatients was 40 years (±16) and average hospital stay was 4 days (±7). Immigrants showed a lower tendency to hospitalisation than Italians with a decreasing trend from 2010 to 2018, contrary to the Italians. The Romanian population has a hospitalisation risk for alcohol abuse/drugs and induced organic mental disorders about twice as much as the Italian population. Women from Morocco experienced caesarean section with complications about four times more than the Italian women. the analysis of routine data highlighted excesses of hospitalisation by some causes and birthplaces. A real-world approach can be useful and timely to the implementation of preventive strategies for specific diseases in defined local areas.


The association between cigarette smoking and risk of colorectal cancer in a large prospective cohort from the United States.
Many studies have reported a 20% to 60% increase in risk of colorectal cancer associated with active smoking. However, neither the U.S. Surgeon General nor the IARC have classified the relationship as causal because of concern about residual confounding. In a prospective study of 184,187 people followed from 1992 to 2005, we used Cox proportional hazard models to examine the relationship of cigarette smoking to incident colorectal cancer, controlling for screening and multiple known and putative risk factors. Information on smoking and time-varying covariates was updated in 1997, 1999, 2001, and 2003. The incidence of colorectal cancer was significantly higher in current [hazard ratios (HR), 1.27; 95% confidence intervals (CI), 1.06-1.52] and former smokers (HR, 1.23; 95% CI, 1.11-1.36) compared with lifelong nonsmokers in analyses that controlled for 13 covariates, including screening. The relative risk was greatest among current smokers with at least 50 years of smoking (HR, 1.38; 95% CI, 1.04-1.84). Among former smokers, risk of colorectal cancer decreased with greater time since cessation (P trend = 0.0003), and also decreased with earlier age at cessation (P trend = 0.0014). No association was seen among former smokers who had quit before age of 40 years or abstained for 31 years or more. Long-term cigarette smoking is associated with colorectal cancer, even after controlling for screening and multiple other risk factors.


Hemodynamic and renal functional changes after acute unilateral nephrectomy in the dog: role of carotid sinus baroreceptors.
We examined the role of the carotid sinus baroreceptors in the hemodynamic and renal functional changes initiated by acute unilateral nephrectomy (AUN). In 15 anesthetized dogs, AUN caused increases in sodium (UNaV) and potassium excretion (UKV) without significant changes in GFR, renal blood flow, or mean arterial pressure. Cardiac output (CO) was measured in five of these animals and decreased from 2.5 +/- 0.2 to 1.9 +/- 0.3 liter/min as total peripheral resistance (TPR) increased. In 11 dogs with bilateral acute surgical carotid sinus denervation, AUN failed to cause any change in renal hemodynamics, UNaV, or UKV that could be distinguished from sham-operated time controls. CO was measured in seven of these dogs and also did not change. AUN caused an immediate, transient elevation in blood pressure that returned to control levels 15 min after nephrectomy. When bilateral carotid occlusion (BCO) was carried out prior to AUN in animals with intact carotid sinus baroreceptors, AUN again caused an immediate increase in blood pressure but no changes in CO or electrolyte excretion. In separate studies, AUN increased UNaV and UKV significantly; when BCO was then carried out after AUN, cation excretion remained elevated. In seven dogs undergoing continuous perfusion of the carotid circulation bilaterally at constant pressure, AUN again failed to increase cation excretion despite a 30 mmHg rise in arterial blood pressure. These studies demonstrate that AUN fails to initiate hemodynamic or compensatory renal functional changes in dogs with bilateral carotid sinus denervation or in animals in which intact baroreceptors are shielded from the systemic circulation by BCO or perfusion at constant pressure, indicating the importance of these baroreceptors in the early changes after AUN.


Characterization of human mutations in phosphorylatable amino acids of the cytosolic regulatory tail of SLC9A1.
The NHE1 isoform of the mammalian Na(+)/H(+) exchanger is a ubiquitous plasma membrane protein that regulates intracellular pH in cells by removing one intracellular proton in exchange for one extracellular sodium. Genetic defects in NHE1 have been shown to affect the growth and motor ability of mice, but mutations in humans have not been studied. NHE1 has a cytosolic C-terminal regulatory domain of approximately 300 amino acids. We investigated the functional effects of two human mutations found in the regulatory phosphorylatable amino acids Ser(703) and Ser(771). A Ser703Pro mutant protein had essentially the same activity, expression, and targeting as the wild type NHE1 protein. In contrast, the Ser771Pro protein had reduced activity and expression of NHE1 protein, though cell surface targeting was normal. In dual pulse assays the Ser771Pro mutant was not further activated by sustained intracellular acidosis but displayed an unusual activation by brief pulses of acidosis. The results demonstrate that the Ser771Pro human genetic mutation has significant and detrimental physiological effects on the activity of the NHE1 protein, SLC9A1.


Alcohol exposure and paracetamol-induced hepatotoxicity.
Most instances of hepatotoxicity due to paracetamol in the United Kingdom and Australia are the result of large overdoses of the drug taken with suicidal or parasuicidal intent. In contrast, serious hepatotoxicity at recommended or near-recommended doses for therapeutic purposes has been reported, mainly from the United States and in association with chronic alcohol use, leading to the widely held belief that chronic alcoholics are predisposed to paracetamol-related toxicity at relatively low doses. Yet the effects of alcohol on paracetamol metabolism are complex. Studies performed in both experimental animals and humans indicate that chronic alcohol use leads to a short-term, two- to threefold increase in hepatic content of cytochrome P4502E1, the major isoform responsible for the generation of the toxic metabolite from paracetamol, although increased oxidative metabolism of paracetamol at recommended doses has not been demonstrated clinically. A reduced hepatic content of glutathione, required to detoxify the reactive metabolite, has been documented in chronic alcoholics, due probably to associated fasting and malnutrition, providing a metabolic basis for any possible predisposition of this group to hepatotoxicity at relatively low paracetamol doses. Simultaneous alcohol and paracetamol ingestion reduces oxidative metabolism of paracetamol in both rodents and humans, predominantly as a consequence of depletion in cytosol of free NADPH. The possibilities that chronic alcohol use may predispose to paracetamol-related hepatotoxicity and that alcohol taken with paracetamol may protect against it, based on these metabolic observations, are examined in this review.


The FDA report on intraocular lenses.
Clinical studies of intraocular lenses (IOLs) as investigational devices have been regulated in the United States by the Food and Drug Administration (FDA) since 9 February 1978. As of August 1982, data have been collected on more than one million IOLs implanted. During the last 12 months of the study, 409 000 IOLs were implanted. Visual acuity of 20/40 or better at one year after surgery was present in 85% of over 45 000 cases reviewed. Increasing patient age, surgical problems, postoperative complications, and adverse reactions were factors that reduced the visual acuity. The current trend in the USA is for implantation of posterior chamber and anterior chamber IOLs.


Neural extrapolation of motion for a ball rolling down an inclined plane.
It is known that humans tend to misjudge the kinematics of a target rolling down an inclined plane. Because visuomotor responses are often more accurate and less prone to perceptual illusions than cognitive judgments, we asked the question of how rolling motion is extrapolated for manual interception or drawing tasks. In three experiments a ball rolled down an incline with kinematics that differed as a function of the starting position (4 different positions) and slope (30°, 45° or 60°). In Experiment 1, participants had to punch the ball as it fell off the incline. In Experiment 2, the ball rolled down the incline but was stopped at the end; participants were asked to imagine that the ball kept moving and to punch it. In Experiment 3, the ball rolled down the incline and was stopped at the end; participants were asked to draw with the hand in air the trajectory that would be described by the ball if it kept moving. We found that performance was most accurate when motion of the ball was visible until interception and haptic feedback of hand-ball contact was available (Experiment 1). However, even when participants punched an imaginary moving ball (Experiment 2) or drew in air the imaginary trajectory (Experiment 3), they were able to extrapolate to some extent global aspects of the target motion, including its path, speed and arrival time. We argue that the path and kinematics of a ball rolling down an incline can be extrapolated surprisingly well by the brain using both visual information and internal models of target motion.


Enantiomer separation of triazole fungicides by high-performance liquid chromatography.
Enantiomer separation is one of the most important prerequisites for the investigation of environmental enantioselective behaviors for chiral pesticides. In the present study, the enantiomer separation of 7 triazole fungicides, i.e., hexaconazole (1), triadimefon (2), tebuconazole (3), diniconazole (4), flutriafol (5), propiconazole (6), and difenoconazole (7), were evaluated using normal phase high-performance liquid chromatography. Chrialcel OD column and Chrialcel OJ column were used. The influence of column temperature was studied for the optimization of the resolution as well as the type and percentage of organic modifier in the mobile phase. The retention factors for the enantiomers of all investigated compounds decreased as the temperature increased. The natural logarithms of the selectivity factors (lnalpha) of hexaconazole (1), tebuconazole (3), flutriafol (5), propiconazole (6) and difenoconazole (7) depended linearly on the inverse of temperature (1/T) while the corresponding values for triadimefon (2) and diniconazole (4) kept unchanged in the studied temperature range 10-35 degrees C. Van't Hoff plots afforded thermodynamic parameters, such as the apparent change in enthalpy DeltaH degrees , the apparent change in entropy DeltaS degrees and the apparent change in DeltaDeltaH degrees and DeltaDeltaS degrees . The thermodynamic parameters (DeltaH degrees , DeltaS degrees , DeltaDeltaH degrees and DeltaDeltaS degrees ) were calculated in order to provide an understanding of the thermosynamic driving forces for enantioseparation. The established method shows perspective to be used for preparing micro-scale amount of pure enantiomers of the chiral triazoles studied.


[Modified Mathieu technic for hypospadias with moderate chordae].
A modified Mathieu technique for hypospadias with moderate chordee is presented. The modifications consist of a skin flap with a large pedicle derived from the dartos fascia, correction of the chordee by resection of the fibrous tissue and dorsal plication of the tunica albuginea and possible reconstruction of the prepuce. Defects of up to 2.5 cm long can be corrected with this modified technique. Seventy six children were operated according to this technique, with a follow-up of more than 24 months in 20 cases. The complications observed in these 20 patients were one fistula, two meatal strictures and one total loss of the skin flap due to infection.


[Natural-valve endocarditis in patients wearing pacemakers].
The presence of infective endocarditis with appearance of vegetations over the tricuspid valve in patients with permanent pacemaker is an uncommon complication. We report here three patients with permanent cardiac pacemakers who developed a picture of infective endocarditis over the electrode with appearance of vegetations over the native valve. The microorganism recovered from blood cultures in the three patients was Staphylococcus. The presence of retained pacemaker leads and repetitive surgical procedures over the pacemaker pouch are two risk factors reported in literature also present in our patients. Infective endocarditis is an uncommon difficult-to-diagnose complication, with a poor prognosis and which requires the removal of the whole infected material, with implantation of an epidermal pacemaker. Some diagnostic and therapeutic issues are discussed.


Efficacy of caspofungin, a 1,3-β-D-glucan synthase inhibitor, on Pneumocystis carinii pneumonia in rats.
Pneumocystis carinii pneumonia (PcP) is a common and potentially fatal opportunistic infection in immunosuppressed patients, and the standard trimethoprim-sulfamethoxazole (TMP-SMZ) treatment has serious side effects. The cell wall of the causative fungal pathogen is enriched in 1-3-β-D-glucan, providing an alternative therapeutic target. We directly compared the efficacy of the 1,3-β-D-glucan synthase inhibitor caspofungin to TMP-SMZ for promoting survival and reducing lung cyst number during the early phase of treatment in a rat model of PcP. Rats were immunosuppressed using dexamethasone for 8 weeks and PcP infection confirmed in test animals by lung print smear. The remaining rats were randomly divided into three control groups, a baseline group and two observed for 7 or 14 days, two caspofungin groups treated intravenously for 7 or 14 days (1 mg/kg/d), and 2 TMP-SMZ positive control groups treated by oral gavage for 7 or 14 days (300 mg/kg/d). Mortality was markedly reduced by both caspofungin and TMP-SMZ after 14 days (caspofungin: 20.0%, TMP-SMZ: 13.3%, Control: 40.0%). Body weight gain in caspofungin-treated rats after 7 (3.04 ± 3.54%) and 14 (4.27 ± 2.79%) days was similar to that in TMP-SMZ-treated rats (3.35 ± 1.88% and 5.85 ± 2.78%, respectively), whereas untreated controls showed weight loss. Lung weight to body weight ratio, and mean cyst number per 50 microscopic fields were significantly lower (all P < 0.05) in caspofungin-treated rats than untreated controls at both 7 and 14 days, and similar to those in the TMP-SMZ-treated rats (all P > 0.05 vs. caspofungin). Caspofungin exhibited similar efficacy to TMP-SMZ for enhancing survival and reducing lung edema and cyst load in a rat model of PcP, suggesting potential clinical utility against PcP.


Seasonality and temporal clustering of Kawasaki syndrome.
The distribution of a syndrome in space and time may suggest clues to its etiology. The cause of Kawasaki syndrome, a systemic vasculitis of infants and children, is unknown, but an infectious etiology is suspected. Seasonality and clustering of Kawasaki syndrome cases were studied in Japanese children with Kawasaki syndrome reported in nationwide surveys in Japan. Excluding the years that contained the 3 major nationwide epidemics, 84,829 cases during a 14-year period (1987-2000) were analyzed. To assess seasonality, we calculated mean monthly incidence during the study period for eastern and western Japan and for each of the 47 prefectures. To assess clustering, we compared the number of cases per day (daily incidence) with a simulated distribution (Monte Carlo analysis). Marked spatial and temporal patterns were noted in both the seasonality and deviations from the average number of Kawasaki syndrome cases in Japan. Seasonality was bimodal with peaks in January and June/July and a nadir in October. This pattern was consistent throughout Japan and during the entire 14-year period. Some years produced very high or low numbers of cases, but the overall variability was consistent throughout the entire country. Temporal clustering of Kawasaki syndrome cases was detected with nationwide outbreaks. Kawasaki syndrome has a pronounced seasonality in Japan that is consistent throughout the length of the Japanese archipelago. Temporal clustering of cases combined with marked seasonality suggests an environmental trigger for this clinical syndrome.


Clinical experience with intravenous zanamivir under an Emergency IND program in the United States (2011-2014).
Since the emergence of 2009 H1N1 virus, intravenous (IV) zanamivir has been authorized as an investigational treatment for patients with serious and life-threatening influenza through an Emergency Investigational New Drug application (EIND). This review encompasses the FDA's EIND database from May 2011 to June 2014. This is a retrospective descriptive review of patient clinical data in the FDA's IV zanamivir EIND database from May 2011 to June 2014. Of 364 IV zanamivir EIND requests, most (83%) patients were aged 18-64 years, 8 (2%) were pregnant, and 29 (8%) were children. 234 (64%) patients had ≥1 comorbidity reported. The majority (87%) were receiving oseltamivir when IV zanamivir was requested, and 33% had suspected (n=120; no improvement or worsening on oseltamivir) H275Y oseltamivir resistance. Influenza A was reported for 300 patients: confirmed 2009 H1N1 (n=163), suspected 2009 H1N1 (n=8), confirmed H3N2 (n=4) and not subtyped (n=125). Influenza B was reported for 25 patients. Many patients (87%) required invasive mechanical ventilation, 23 (6%) received high frequency oscillatory ventilation, and 74 (20%) received extracorporeal membrane oxygenation (ECMO). 289 (79%) patients had ≥1 complication such as renal failure (n=124; 77/124 required dialysis), bacteraemia (n=18), shock (n=95) or pneumonia (n=159). Of 134 (37%) patients with available outcome data, 83 died and 51 survived. IV zanamivir EIND authorizations were for treatment of critically ill adult patients with 2009 H1N1, including a substantial number with suspected oseltamivir resistance. Data from prospective, randomized controlled trials are needed and are ongoing to assess the safety and efficacy of IV zanamivir for treatment of hospitalized patients with severe influenza.


Effects of Training and Competition Load on Neuromuscular Recovery, Testosterone, Cortisol, and Match Performance During a Season of Professional Football.
Introduction: Training load and other measures potentially related to match performance are routinely monitored in team-sport athletes. The aim of this research was to examine the effect of training load on such measures and on match performance during a season of professional football. Materials and Methods: Training load was measured daily as session duration times perceived exertion in 23 A-League football players. Measures of exponentially weighted cumulative training load were calculated using decay factors representing time constants of 3-28 days. Players performed a countermovement jump for estimation of a measure of neuromuscular recovery (ratio of flight time to contraction time, FT:CT), and provided a saliva sample for measurement of testosterone and cortisol concentrations 1-day prior to each of 34 matches. Match performance was assessed via ratings provided by five coaching and fitness staff on a 5-point Likert scale. Effects of training load on FT:CT, hormone concentrations and match performance were modeled as quadratic predictors and expressed as changes in the outcome measure for a change in the predictor of one within-player standard deviation (1 SD) below and above the mean. Changes in each of five playing positions were assessed using standardization and magnitude-based inference. Results: The largest effects of training were generally observed in the 3- to 14-day windows. Center defenders showed a small reduction in coach rating when 14-day a smoothed load increased from -1 SD to the mean (-0.31, ±0.15; mean, ±90% confidence limits), whereas strikers and wide midfielders displayed a small increase in coach rating when load increased 1 SD above the mean. The effects of training load on FT:CT were mostly unclear or trivial, but effects of training load on hormones included a large increase in cortisol (102, ±58%) and moderate increase in testosterone (24, ±18%) in center defenders when 3-day smoothed training load increased 1 SD above the mean. A 1 SD increase in training load above the mean generally resulted in substantial reductions in testosterone:cortisol ratio. Conclusion: The effects of recent training on match performance and hormones in A-League football players highlight the importance of position-specific monitoring and training.


HIF-2α mediates hypoxia-induced LIF expression in human colorectal cancer cells.
Leukemia inhibitory factor (LIF), a multi-functional cytokine, has a complex role in cancer. While LIF induces the differentiation of several myeloid leukemia cells and inhibits their growth, it also promotes tumor progression, metastasis and chemoresistance in many solid tumors. LIF is frequently overexpressed in a variety of human tumors and its overexpression is often associated with poor prognosis of patients. Currently, the mechanism for LIF overexpression in tumor cells is not well-understood. Here, we report that hypoxia, a hallmark of solid tumors, induced LIF mRNA expression in human colorectal cancer cells. Analysis of LIF promoter revealed several hypoxia-responsive elements (HREs) that can specifically interact with and be transactivated by HIF-2α but not HIF-1α. Consistently, ectopic expression of HIF-2α but not HIF-1α transcriptionally induced LIF expression levels in cells. Knockdown of endogenous HIF-2α but not HIF-1α by siRNA largely abolished the induction of LIF by hypoxia in cells. Furthermore, there is a strong association of HIF-2α overexpression with LIF overexpression in human colorectal cancer specimens. In summary, results from this study demonstrate that hypoxia induces LIF expression in human cancer cells mainly through HIF-2α, which could be an important underlying mechanism for LIF overexpression in human cancers.


Reconstruction of Rayleigh-Lamb dispersion spectrum based on noise obtained from an air-jet forcing.
The time-domain cross correlation of incoherent and random noise recorded by a series of passive sensors contains the impulse response of the medium between these sensors. By using noise generated by a can of compressed air sprayed on the surface of a plexiglass plate, we are able to reconstruct not only the time of flight but the whole wave forms between the sensors. From the reconstruction of the direct A(0) and S(0) waves, we derive the dispersion curves of the flexural waves, thus estimating the mechanical properties of the material without a conventional electromechanical source. The dense array of receivers employed here allow a precise frequency-wavenumber study of flexural waves, along with a thorough evaluation of the rate of convergence of the correlation with respect to the record length, the frequency, and the distance between the receivers. The reconstruction of the actual amplitude and attenuation of the impulse response is also addressed in this paper.


Treatment of inflammatory bowel disease associated E. coli with ciprofloxacin and E. coli Nissle in the streptomycin-treated mouse intestine.
E. coli belonging to the phylogenetic group B2 are linked to Inflammatory Bowel Disease (IBD). Studies have shown that antimicrobials have some effect in the treatment of IBD, and it has been demonstrated that E. coli Nissle has prophylactic abilities comparable to 5-aminosalicylic acid (5-ASA) therapy in ulcerative colitis. The objective of this study was to test if ciprofloxacin and/or E. coli Nissle could eradicate IBD associated E. coli in the streptomycin-treated mouse intestine. After successful colonization with the IBD associated E. coli strains in mice the introduction of E. coli Nissle did not result in eradication of either IBD associated strains or an E. coli from a healthy control, instead, co-colonization at high levels were obtained. Treatment of mice, precolonized with IBD associated E. coli, with ciprofloxacin for three days alone apparently resulted in effective eradication of tested E. coli. However, treatment of precolonized mice with a combination of ciprofloxacin for 3 days followed by E. coli Nissle surprisingly allowed one IBD associated E. coli to re-colonize the mouse intestine, but at a level 3 logs under E. coli Nissle. A prolonged treatment with ciprofloxacin for 7 days did not change this outcome. In the mouse model E. coli Nissle can not be used alone to eradicate IBD associated E. coli; rather, 3 days of ciprofloxacin are apparently efficient in eradicating these strains, but surprisingly, after ciprofloxacin treatment (3 or 7 days), the introduction of E. coli Nissle may support re-colonization with IBD associated E. coli.


Understanding mechanisms underlying the pathology of immune reconstitution inflammatory syndrome (IRIS) by using animal models.
Despite the increasing number of clinical reports on immune reconstitution inflammatory syndrome (IRIS), mechanistic understanding of IRIS is still largely limited. The main focus of this review is to summarize animal studies, which were performed to better understand the cellular and molecular mechanisms underlying the pathology of IRIS. Three IRIS animal models have been reported. They are Mycobacterial IRIS (M-IRIS), cryptococcal IRIS (C-IRIS) and Pneumocystis-IRIS. M-IRIS animal model suggested that, rather than lymphopenia itself, the failure to clear the pathogen by T cells results in excessive priming of the innate immune system. If this happens before T cell reconstitution, hosts likely suffer IRIS upon T cell reconstitution. Interestingly, T cells specific to self-antigens, not only pathogen-specific, could drive IRIS as well. The mechanism to develop IRIS is quite complicated, including multiple layers of host immune responses; the innate immune system that detects pathogens and prime host immunity, and the adaptive immune system that is reconstituted but hyper-activated particularly through CD4+ T cells. Animal models of IRIS, although there are still small numbers of studies available, have already provided significant insights on the mechanistic understanding of IRIS.