Quantifying Exposure and Intra-Individual Reliability of High-Speed and Sprint Running During Sided-Games Training in Soccer Players: A Systematic Review and Meta-analysis.
Sided games (i.e., small sided, medium sided, large sided) involve tactical, technical, physical, and psychological elements and are commonly implemented in soccer training. Although soccer sided-games research is plentiful, a meta-analytical synthesis of external load exposure during sided games is lacking. The objective of this systematic review and meta-analysis was to: (1) synthesize the evidence on high-speed and sprint running exposure induced by sided games in adult soccer players, (2) establish pooled estimates and intra-individual reliability for high-speed and sprint running exposure, and (3) explore the moderating effects of game format and playing constraints. A literature search was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 guidelines. Four databases (PubMed/MEDLINE, Scopus, SPORTDiscus, Web of Science Core Collection) were systematically searched up to 25 January, 2022. Eligibility criteria were adult soccer players (population); training programs incorporating sided games (intervention); game manipulations including number of players, pitch dimension, and game orientation (comparator); and high-speed, very high-speed, and sprint relative (m[Formula: see text]min-1) running distances and associated intra-individual reliability (outcome). Eligible study risk of bias was evaluated using RoBANS. Pooled estimates for high-speed and sprint running exposure, and their intra-individual reliability, along with the moderating effect of tracking device running velocity thresholds, pitch dimension (i.e., area per player), and game orientation (i.e. score or possession), were determined via a multi-level mixed-effects meta-analysis. Estimate uncertainty is presented as 95% compatibility intervals (CIs) with the likely range of relative distances in similar future studies determined via 95% prediction intervals. A total of 104 and 7 studies met our eligibility criteria for the main and reliability analyses, respectively. The range of relative distances covered across small-sided games, medium-sided games, and large-sided games was 14.8 m[Formula: see text]min-1 (95% CI 12.3-17.4) to 17.2 m[Formula: see text]min-1 (95% CI 13.5-20.8) for high-speed running, 2.7 m[Formula: see text]min-1 (95% CI 1.8-3.5) to 3.6 m[Formula: see text]min-1 (95% CI 2.3-4.8) for very high-speed running, and 0.2 m[Formula: see text]min-1 (95% CI 0.1-0.4) to 0.7 m[Formula: see text]min-1 (95% CI 0.5-0.9) for sprinting. Across different game formats, 95% prediction intervals showed future exposure for high-speed, very high-speed running, and sprinting to be 0-46.5 m[Formula: see text]min-1, 0-14.2 m[Formula: see text]min-1, and 0-2.6 m[Formula: see text]min-1, respectively. High-speed, very high-speed running, and sprinting showed poor reliability with a pooled coefficient of variation of 22.8% with distances being moderated by device speed thresholds, pitch dimension, and game orientation. This review is the first to provide a detailed synthesis of exposure and intra-individual reliability of high-speed and sprint running during soccer sided games. Our estimates, along with the moderating influence of common programming variables such as velocity thresholds, area per player, and game orientation should be considered for informed planning of small-sided games, medium-sided games, and large-sided games soccer training. Open Science Framework available through https://osf.io/a4xr2/ .


Regulation of ribosomal S6 protein kinase-p90(rsk), glycogen synthase kinase 3, and beta-catenin in early Xenopus development.
beta-Catenin is a multifunctional protein that binds cadherins at the plasma membrane, HMG box transcription factors in the nucleus, and several cytoplasmic proteins that are involved in regulating its stability. In developing embryos and in some human cancers, the accumulation of beta-catenin in the cytoplasm and subsequently the nuclei of cells may be regulated by the Wnt-1 signaling cascade and by glycogen synthase kinase 3 (GSK-3). This has increased interest in regulators of both GSK-3 and beta-catenin. Searching for kinase activities able to phosphorylate the conserved, inhibitory-regulatory GSK-3 residue serine 9, we found p90(rsk) to be a potential upstream regulator of GSK-3. Overexpression of p90(rsk) in Xenopus embryos leads to increased steady-state levels of total beta-catenin but not of the free soluble protein. Instead, p90(rsk) overexpression increases the levels of beta-catenin in a cell fraction containing membrane-associated cadherins. Consistent with the lack of elevation of free beta-catenin levels, ectopic p90(rsk) was unable to rescue dorsal cell fate in embryos ventralized by UV irradiation. We show that p90(rsk) is a downstream target of fibroblast growth factor (FGF) signaling during early Xenopus development, since ectopic FGF signaling activates both endogenous and overexpressed p90(rsk). Moreover, overexpression of a dominant negative FGF receptor, which blocks endogenous FGF signaling, leads to decreased p90(rsk) kinase activity. Finally, we report that FGF inhibits endogenous GSK-3 activity in Xenopus embryos. We hypothesize that FGF and p90(rsk) play heretofore unsuspected roles in modulating GSK-3 and beta-catenin.


Proteins involved in building, maintaining and remodeling of yeast cell walls.
The cell wall defines the shape and provides osmotic stability to the yeast cell. It also serves to anchor proteins required for communication of the yeast cell with surrounding molecules and other cells. It is synthesized as a complex structure with β-1,3-glucan chains forming the basic network to which β-1,6-glucan, chitin and a number of mannoproteins are attached. Synthesis, maintaining and remodeling of this complex structure require a set of different synthases, hydrolases and transglycosidases whose concerted activities provide necessary firmness but at the same time flexibility of the wall moiety. The present state of comprehension of the interplay of these proteins in the yeast cell wall is the subject of this article.


Mapping genomic features of tiling microarray data by TileMapper.
The recent revolution of genomics techniques has allowed the detection of various sequence features and biological variations on whole-genome scale. However, these high-resolution data present significant challenges for experimental biologists to understand and analyze. The conventional way is to use genome browsers to locate and visualize regions of interest. But it lacks user-friendly data mining functionality. Here we present a protocol that allows rapid annotation of genomic coordinate data by using TileMapper. Interesting biological annotations from large-scale genomic data, such as transcriptome analysis, chromatin immunoprecipitation on chip, or methyl-DNA immunoprecipitation (MeDIP) studies generated from the tiling microarrays and other platforms, could be analyzed without requiring computational skills. The outputs are saved in tabulated format, which permit flexible and simple processing in spreadsheet software, or to be exported to other pipelines for subsequent analysis.


A rapid method for 90Sr determination in the presence of 137Cs in environmental samples.
A rapid method for the determination of 90Sr in the presence of 137Cs using the Cherenkov radiation technique is described. The contribution of 137Cs to gross Cherenkov radiation (90Y + 137Cs) was examined for 137Cs /90Sr ratios ranging from 0.09 to 2.50 for 137Cs activities ranging from 2 to 1,211 Bq. Results from direct Cherenkov radiation measurement and results after radiochemical separation of 90Y from 90Sr for samples containing both 90Sr and 137Cs were compared. Errors below 5% were obtained for 137Cs /90Sr ratios lower than 1, when no separation was performed, independently of the activity level. However, errors between 10% and 35% were obtained for 137Cs/90Sr ratios higher than 1. In order to determine 90Sr activity in the presence of 137Cs using the Cherenkov technique, a multiple linear regression analysis model was established to correct the data for 137Cs content. The mathematical correction proposed was validated using 66 artificially contaminated lettuce samples in a laboratory experiment by taking into account the activity levels of 137Cs and 90Sr and the radionuclide ratios. Comparison of mathematically corrected radionuclide ratios with the results obtained without correction shows that, for radionuclide ratios higher than 1, error values for measuring 90Sr activity using the mathematical model were much smaller than when no radiochemical separation was performed. On the other hand, for ratios lower than 1, error values when measuring 90Sr activity with radiochemical separation were smaller than when mathematical correction was performed. In spite of this, the mathematical correction is an appropriate way of reducing the time needed to determine radiostrontium using the Cherenkov radiation technique. The method proposed could be a powerful tool for environmental research whenever the contents of 90Sr and 137Cs have to be determined.


Impact of older age in patients receiving atezolizumab and bevacizumab for hepatocellular carcinoma.
Combination atezolizumab/bevacizumab is the gold standard for first-line treatment of unresectable hepatocellular carcinoma (HCC). Our study investigated the efficacy and safety of combination therapy in older patients with HCC. 191 consecutive patients from eight centres receiving atezolizumab and bevacizumab were included. Overall survival (OS), progression-free survival (PFS), overall response rate (ORR) and disease control rate (DCR) defined by RECIST v1.1 were measured in older (age ≥ 65 years) and younger (age < 65 years) age patients. Treatment-related adverse events (trAEs) were evaluated. The elderly (n = 116) had higher rates of non-alcoholic fatty liver disease (19.8% vs. 2.7%; p < .001), presenting with smaller tumours (6.2 cm vs 7.9 cm, p = .02) with less portal vein thrombosis (31.9 vs. 54.7%, p = .002), with fewer patients presenting with BCLC-C stage disease (50.9 vs. 74.3%, p = .002). There was no significant difference in OS (median 14.9 vs. 15.1 months; HR 1.15, 95% CI 0.65-2.02 p = .63) and PFS (median 7.1 vs. 5.5 months; HR 1.11, 95% CI 0.54-1.92; p = .72) between older age and younger age. Older patients had similar ORR (27.6% vs. 20.0%; p = .27) and DCR (77.5% vs. 66.1%; p = .11) compared to younger patients. Atezolizumab-related (40.5% vs. 48.0%; p = .31) and bevacizumab-related (44.8% vs. 41.3%; p = .63) trAEs were comparable between groups. Rates of grade ≥3 trAEs and toxicity-related treatment discontinuation were similar between older and younger age patients. Patients 75 years and older had similar survival and safety outcomes compared to younger patients. Atezolizumab and bevacizumab therapy is associated with comparable efficacy and tolerability in older age patients with unresectable HCC.


rpoB mutation conferring rifampin resistance in Streptococcus pyogenes.
Streptococcus pyogenes BM4478 and Staphylococcus aureus BM4479 were isolated from a patient undergoing rifampin therapy. High-level resistance to rifampin was due to the following mutations in the rpoB gene: Ser522Leu in strain BM4478 and His526Asn and Ser574Leu in strain BM4479.


[A clinical trial of oxymatrine in treating chronic viral hepatitis type B].
To investigate the therapeutic effect of oxymatrine on chronic viral hepatitis type B. 303 patients were randomly allocated either to a treatment group or a control group. The treatment group consisted of 253 patients treated with intravenous or intra-muscular injection of oxymatrine and oral oxymatrine capsule. Oral tiopronin was used in the control group. At the end of treatment, the rate of normal ALT was similar among the different groups. The rate of normal ALT was 53.3% - 58.3% in the three oxymatrine groups six months after the end of treatment. It was higher than that of the tiopronin group (P < 0.05). After a follow up of six months, the rate of negative HBeAg was 30.0% - 40.9% in the three oxymatrine groups. It was higher than that of the tiopronin group (16.7%). The rate of negative HBV DNA was 39.2% - 49.5% in the three oxymatrine groups. It was also higher than that of the tiopronin group. Oxymatrine can improve the liver function and increase the negative rate of HBeAg and HBV DNA in the patients with chronic hepatitis B. The therapeutic effect of oxymatrine will persist after the cessation of administration.


Combined approach for extensive maxillectomy: technique and cadaveric dissection.
Currently described endoscopic techniques for subtotal resections of the maxilla include endoscopic medial maxillectomy and extended endoscopic medial maxillectomy; however, a complete resection of the maxilla is sometimes warranted. We describe a combined transoral and endoscopic technique for total and subtotal maxillectomy in an attempt to decrease the morbidity of traditional approaches. Technical note, Feasibility, Human cadaveric dissection. Ten total and subtotal maxillectomies were performed in human specimens without the need of facial incisions or transfixion of the nasal septum. The pterygopalatine and infratemporal fossas were accessed and dissected in all cases. A combined transoral and endoscopic approach is feasible and can be used in selected patients when other minimally endoscopic techniques are not indicated. The benefits of no facial incisions and/or transfixion of the nasal septum, potential improvement in hemostasis, and visual magnification may help to decrease the morbidity of traditional open approaches.


Immunological treatment of multiple sclerosis.
Immunosuppressive treatment of multiple sclerosis (MS) is based on the autoimmune hypothesis for which the main evidence is the close histological similarity between the human disease and chronic relapsing EAE. Although controlled trials indicate that ACTH is effective in accelerating recovery from relapses, long term ACTH or oral steroids are ineffective. Two controlled trials have suggested a beneficial effect of azathioprine, but neither was conducted "blind" and neither was sufficiently convincing to cause the widespread adoption of azathioprine by neurologists. One controlled trial, also not blind, reported a beneficial effect of an intensive course of cyclophosphamide, but this hazardous treatment will not be widely adopted unless other trials confirm this result. The converse hypothesis that MS is due to a deficient immune response to a virus has led to trials of immunostimulation. Interferon and levamisole have proven ineffective so far, but transfer factor slowed disease progression in one well conducted trial.


Salmonella newport infections transmitted by fiberoptic colonoscopy.
During a 2-week period following the colonoscopy and biopsy of a patient with acute Salmonella newport gastroenteritis, S. newport was recovered from colonic aspirates or fecal specimens of eight of 28 patients from whom specimens were cultured during or after colonoscopy. Two of the eight persons from whom S. newport was isolated developed acute gastroenteritis, two had asymptomatic infections, and four had positive aspirates collected through a colonoscope but did not become infected. Although S. newport was never recovered from the four colonoscopes used during the outbreak, cultures of one of the colonic biopsy forceps grew S. newport. Contamination of the equipment most likely occurred during colonoscopy of the index patient. Inadequate disinfection of the equipment allowed the organism to survive and possibly to cross-contaminate other colonoscopes, and the organism was then transmitted to other patients by use of the contaminated colonoscopes or the contaminated biopsy forceps. Implemented control measures terminated the outbreak.


Cross-bridge cooperativity during isometric contraction and unloaded shortening of skeletal muscle.
Whether the two heads of skeletal muscle myosin work independently or cooperatively remains an open question in muscle biophysics. While individual myosin heads are sufficient for ATPase activity (Reisler (1980) J Mol Biol 138: 93-107) and force production (Harada et al. (1987) Nature 326: 805-808), it has also been reported that in situ, the two heads of a myosin molecule work cooperatively (Chaen et al. (1986) J Biol Chem 261(29): 13,632-13,636). To examine the role of cross-bridge cooperativity on isometric contraction and unloaded shortening we progressively inactivated myosin cross-bridges via titration with para-phenylenedimaleimide. The resting fiber ATPase was measured to provide an estimate of the fraction of active cross-bridges remaining during the titration. Isometric force and unloaded shortening velocity decline more rapidly than the resting ATPase as the titration proceeds. This is inconsistent with models for independent force generation and suggests cooperative action of myosin cross-bridges when muscle is isometrically contracting or shortening under zero load. However the degree of cooperativity depends on the type of muscle activity. While isometric force declines in a manner consistent with pair-wise cooperative action of myosin heads, unloaded shortening velocity declines more rapidly (greater cooperativity). Therefore, myosin cross-bridges in situ may be capable of at least two types of cooperative interactions, pair-wise cooperativity (when isometric) and another form of cooperativity that is sensitive to longer range interactions transmitted from other cross-bridges in the ensemble (during unloaded shortening).


Ultrastructure of Cryptococcus neoformans in the cerebrospinal fluid of a patient with cryptococcal meningitis.
The ultrastructure of Cryptococcus neoformans in the cerebrospinal fluid, which was obtained from a patient suffering from systemic lupus erythematosus and cryptococcal meningitis, before treatment and on the 10th and 20th day after the start of treatment was studied. Numerous cryptococci were detected in their cerebrospinal fluid before treatment. However, in most of them their biological activity was low and their organelles were not clear. A few yeasts preserved well their organelles. Such cells showed a tendency to develop vesicular membrane structures (lomasomes) touching the internal part of the cell wall. In the cerebrospinal fluid on the 20th day all of the yeasts were dead, even though numerous yeasts were observed by an India-ink method. At this time no colonies were recovered from the fluid.


Delayed luminescence from bovine Achilles' tendon and its dependence on collagen structure.
Delayed luminescence from ox Achilles' tendons is found to depend on the order parameters of the collagen fibres and on both water content and the temperature, which affect those parameters.


Collaborative evaluation within a framework of stakeholder-oriented evaluation approaches.
Collaborative Evaluation systematically invites and engages stakeholders in program evaluation planning and implementation. Unlike "distanced" evaluation approaches, which reject stakeholder participation as evaluation team members, Collaborative Evaluation assumes that active, on-going engagement between evaluators and program staff, result in stronger evaluation designs, enhanced data collection and analysis, and results that stakeholder understand and use. Among similar "participant-oriented" evaluation approaches (Fitzpatrick, Sanders, & Worthen, 2011), Collaborative Evaluation distinguishes itself in that it uses a sliding scale for levels of collaboration. This means that different program evaluations will experience different levels of collaborative activity. The sliding scale is applied as the evaluator considers each program's evaluation needs, readiness, and resources. While Collaborative Evaluation is a term widely used in evaluation, its meaning varies considerably. Often used interchangeably with participatory and/or empowerment evaluation, the terms can be used to mean different things, which can be confusing. The articles use a comparative Collaborative Evaluation Framework to highlight how from a theoretical perspective, Collaborative Evaluation distinguishes itself from the other participatory evaluation approaches.


Patterns of molecular genetic variation among cat breeds.
Genetic variation in cat breeds was assessed utilizing a panel of short tandem repeat (STR) loci genotyped in 38 cat breeds and 284 single-nucleotide polymorphisms (SNPs) genotyped in 24 breeds. Population structure in cat breeds generally reflects their recent ancestry and absence of strong breed barriers between some breeds. There is a wide range in the robustness of population definition, from breeds demonstrating high definition to breeds with as little as a third of their genetic variation partitioning into a single population. Utilizing the STRUCTURE algorithm, there was no clear demarcation of the number of population subdivisions; 16 breeds could not be resolved into independent populations, the consequence of outcrossing in established breeds to recently developed breeds with common ancestry. These 16 breeds were divided into 6 populations. Ninety-six percent of cats in a sample set of 1040 were correctly assigned to their classified breed or breed group/population. Average breed STR heterozygosities ranged from moderate (0.53; Havana, Korat) to high (0.85; Norwegian Forest Cat, Manx). Most of the variation in cat breeds was observed within a breed population (83.7%), versus 16.3% of the variation observed between populations. The hierarchical relationships of cat breeds is poorly defined as demonstrated by phylogenetic trees generated from both STR and SNP data, though phylogeographic grouping of breeds derived completely or in part from Southeast Asian ancestors was apparent.


Increased preoperative C-reactive protein levels are associated with inhospital death after coronary artery bypass surgery.
Increased C-reactive protein (CRP) is a predictor of cardiovascular risk, but its influence on outcomes after coronary artery bypass grafting surgery (CABG) is still incompletely studied. We studied the association between preoperative CRP and inhospital death after CABG. Patients with acute or chronic infectious or inflammatory disorders, autoimmune diseases, cancer, and prior cardiac surgery were excluded. Seventy-six patients were studied [27.6% with elevated CRP (>3 mg/l)]. Elevated CRP was more frequently found in patients who died than in those who survived (83.3% vs 17.1%, p = 0.003); mean CRP levels were, respectively, 6.5 ± 3.4 vs 2.4 ± 3.5 mg/l (p = 0.03). The hazard ratio of death was 11.7 for elevated CRP, and the ROC curve for the discrimination of death with CRP had an area under the curve of 0.82. An improvement to mortality risk prediction following CABG may be offered by the preoperative analysis of CRP.


Retrospective analysis of pre- and peri-operative imaging in confirmed proximal colonic cancers--possible implications for screening flexible sigmoidoscopy.
Faecal occult blood testing is being introduced for population screening in the United Kingdom. Flexible sigmoidoscopy may provide a viable alternative. The outcomes of the flexible sigmoidoscopy trial are awaited but the most obvious disadvantage is that only the lower third of the colon is examined and proximal pathology cannot be excluded. The relationship between proximal pathology and distal findings at flexible sigmoidoscopy is uncertain. The aim of this study was to determine the incidence of distal neoplasia in patients with confirmed proximal cancers of the colon. All confirmed proximal colonic cancers (defined as those proximal to the splenic flexure) were identified from a database of pathology specimens at a single centre between January 1999 and August 2006. A retrospective analysis of preoperative and peri-operative mucosal imaging (contrast enema, colonoscopy and CT colonography) was conducted to identify any distal neoplasia in these patients. A total of 348 patients were identified. Pre- or peri-operative mucosal imaging was identified in 231 (66%) and 49 (21%) had distal neoplasia. Nineteen (8%) of these patients would have gone on to have a colonoscopy based on the UK flexible sigmoidoscopy trial protocol and 92% of the cohort would not have had a colonoscopy. Nearly 80% of confirmed proximal cancers in our series did not have any demonstrable distal neoplasia. Only 8% of our cohort would have proceeded to colonoscopy. A very significant number of proximal cancers would not have been detected.


Rapid identification of cervical adenocarcinoma and cervical squamous cell carcinoma tissue based on Raman spectroscopy combined with multiple machine learning algorithms.
Cervical cancer has a long latency, and early screening greatly reduces mortality. In this study, cervical adenocarcinoma and cervical squamous cell carcinoma tissue data were collected by Raman spectroscopy, and then, the adaptive iteratively reweighted penalized least squares (airPLS) algorithm and Vancouver Raman algorithm (VRA) were used to subtract the background of the collected data. The following five feature extraction algorithms were applied: partial least squares (PLS), principal component analysis (PCA), kernel principal component analysis (KPCA), isometric feature mapping (isomap) and locally linear embedding (LLE). The k-nearest neighbour (KNN), extreme learning machine (ELM), decision tree (DT), backpropagation neural network (BP), genetic optimization backpropagation neural network (GA-BP) and linear discriminant analysis (LDA) classification models were then established through the features extracted by different feature extraction algorithms. In total, 30 types of classification models were established in this experiment. This research includes eight good models, airPLS-PLS-KNN, airPLS-PLS-ELM, airPLS-PLS-GA-BP, airPLS-PLS-BP, airPLS-PLS-LDA, airPLS-PCA-KNN, airPLS-PCA-LDA, and VRA-PLS-KNN, whose diagnostic accuracy was 96.3 %, 95.56 %, 95.06 %, 94.07 %, 92.59 %, 85.19 %, 85.19 % and 85.19 %, respectively. The experimental results showed that the model established in this article is simple to operate and highly accurate and has a good reference value for the rapid screening of cervical cancer.


Post-operative survival in head and neck cancer patients with elevated troponins.
The strenuous demands of head and neck cancer surgery (HNS) place patients at increased risk of myocardial injury. Troponin positivity (TP) post-operatively is a predictor of increased complications and mortality. The present study is the first to investigate the effects of TP on potential delays in adjuvant treatment and disease-specific survival. All patients undergoing HNS from 2014 to 2016 had troponins measured at a single academic centre. Relevant patient data was extracted on retrospective chart review. The main outcome measures were the impact of TP on timing of adjuvant treatment and disease-specific survival. Of 166 patients, 26 (15.6%) developed TP post-operatively. There was no significant difference between cohorts for baseline characteristics except for age. Overall and disease-specific survival for TP patients were respectively 45.9% and 57.4% at 3 years. There was no significant difference between cohorts for overall and disease-specific survival, and time to adjuvant therapy. No significant association was found between TP and overall and disease-specific survival, and time to adjuvant therapy.


Immunoelectron microscopic localization of serine: pyruvate aminotransferase in rat eosinophile leukocytes.
Localization of serine: pyruvate aminotransferase [EC 188.8.131.52]; SPT in rat eosinophile leukocytes was investigated by protein A-gold technique. Thin sections of rat intestine were incubated with anti-SPT, followed by protein A-gold complex. Labelling with gold particles was seen on the specific granules of eosinophile leukocytes, in which 78% of the gold particles were localized on their paracrystalline cores and 22% on matrix, indicating that the main intragranular sites of SPT are the core. Other cell organelles such as nucleus and mitochondria were not labelled specifically. Quantitative analysis of labelling density in the subcellular compartments also confirmed that SPT is present exclusively in the specific granules.


Cardiomyopathy from 1,1-Difluoroethane Inhalation.
Consumer aerosol products can be inhaled for their psychoactive effects, but with attendant adverse health effects including "sudden sniffing death." Cardiomyopathy has rarely been described in association with 1,1-difluoroethane (DFE), a common aerosol propellant. We report a 33-year-old male who developed acute myocardial injury and global hypokinesis along with rhabdomyolysis, acute kidney injury, and fulminant hepatitis after 2 days' nearly continuous huffing. Workup for other causes, including underlying coronary artery disease, was negative. His cardiac function improved over time. The exact mechanism of DFE's effects is uncertain but may include catecholamine-induced cardiomyopathy, coronary vasospasm, or direct cellular toxicity.


Genome sequencing identifies KMT2E-disrupting cryptic structural variant in a female with O'Donnell-Luria-Rodan syndrome.
We describe a patient from the 100,000 Genomes Project with a complex de novo structural variant within KMT2E leading to O'Donnell-Luria-Rodan syndrome. This case expands the mutational spectrum for this syndrome and highlights the importance of revisiting unsolved cases using better SV prioritisation tools and updated gene panels.


Identification of the ligand trans to thiolate in cytochrome P-450 LM2 by chemical modification.
About 3 tyrosine residues of cytochrome P-450 LM2 are accessible to chemical modification with tetranitromethane. Nitration of two tyrosines inactivates the enzyme to about 20%. The partial formation of a hyper-porphyrin spectrum originating from the pK shift by nitration and formation of a tyrosinate is prevented by modification in the presence of the inhibitor metyrapone. These findings support the assumption of a tyrosine residue as sixth ligand of the heme iron in cytochrome P-450 LM2.


Intrinsic penicillin resistance in penicillinase-producing Neisseria gonorrhoeae strains.
The minimal inhibitory concentrations (MICs) of ampicillin for fifty strains of beta-lactamase-producing Neisseria gonorrhoeae (PPNG) isolated in Japan ranged from 1.56 to 200 micrograms/ml, and all the strains harbored a 4.5 megadalton plasmid. These strains were classified into two groups: dicloxacillin-susceptible (28%) and -resistant group (72%). A linear correlation was found in the dicloxacillin-susceptible strains between their beta-lactamase activity and the susceptibility to ampicillin, but not in the dicloxacillin-resistant strains. This suggests that the high ampicillin resistance in PPNG is due not only to acquiring the beta-lactamase producing plasmid, but also to some intrinsic resistance of the strains. To investigate a cause of the high ampicillin resistance, the beta-lactamase-producing plasmid, pTMS1, was transferred by conjugation to a penicillin-susceptible gonococcal strain as well as to its isogenic multiply antibiotic-resistant transformants, and the susceptibility of the transconjugants to ampicillin was determined. Acquisition of pTMS1 by a penicillin-susceptible strain resulted in a 32-fold increase in resistance to ampicillin, whereas the increase was 128-fold for its isogenic strains which contain some chromosomal mutations. These results suggest that reduced permeability of the outer membrane to ampicillin underlies the high ampicillin resistance of PPNG.


Bis-(salicylaldehyde-benzhydrylimino)nickel complexes with different electron groups: crystal structure and their catalytic properties toward (co)polymerization of norbornene and 1-hexene.
Eight bis-(salicylaldehyde-benzhydrylimino)nickel complexes with different electron groups (Ni1-Ni8), Ni{(3-R1)(5-R2)C6H2(O)CHNCH(C6H5)2}2, (R1 = H, R2 = H, Ni1; R1 = H, R2 = CH3, Ni2; R1 = H, R2 = OCH3, Ni3; R1 = H, R2 = Br, Ni4; R1 = CH3, R2 = H, Ni5; R1 = OCH3, R2 = H, Ni6; R1 = Br, R2 = Br, Ni7; R1 = Cl, R2 = Cl, Ni8), were synthesized and their crystal structures were characterized using single crystal X-ray diffraction. The results revealed that Ni1-Ni6 belong to the monoclinic system (space group P2(1)/n), Ni7 belongs to the monoclinic system (space group C2/c) and Ni8 belongs to the triclinic system (space group P1̄). All nickel complexes exhibited high activities (0.46-2.07 × 106 gpolymer molNi -1 h-1) toward norbornene homopolymerization, and a strong electron-withdrawing group on the salicylaldimino aromatic ring can enhance the catalytic activity and favor polymerization. Ni1 and Ni2 exhibited high activities (0.55-2.40 × 105 gpolymer molNi -1 h-1) toward copolymerization of norbornene and 1-hexene in the presence of B(C6F5)3. The 1-hexene content in the copolymers could be controlled up to 7.98-12.50% by varying the comonomer feed ratio of 1-hexene from 10 to 50%. It is observed that when the 5-position of the salicylaldimino aromatic ring has a substituent (-CH3), the 1-hexene insertion rate is lower than that without a substituent. In addition, the polymers showed high molecular weights (1.5-2.4 × 105 g mol-1) and narrow molecular weight distributions (1.62-1.89). The obtained polymers were also verified to be amorphous copolymers and had high thermal stability, good solubility and optical transparency.


HIV-1 Gag and Vpr impair the inflammasome activation and contribute to the establishment of chronic infection in human primary macrophages.
The successful establishment of HIV-1 infection is related to inflammasome blocking or inactivation, which can result in the viral evasion of the immune responses and formation of reservoirs in several tissues. In this sense, we aimed to evaluate the viral and cellular mechanisms activated during HIV-1 infection in human primary macrophages that allow an effective viral replication in these cells. We found that resting HIV-1-infected macrophages, but not those activated in classical or alternative patterns, released IL-1β and other pro-inflammatory cytokines, and showed increased CXCL10 expression, without changes in the NLRP3, AIM2 or RIG-I inflammasome pathways. Also, similar levels of Casp-1, phosphorylated NF-κB (p65) and NLRP3 proteins were found in uninfected and HIV-1-infected macrophages. Likewise, no alterations were detected in ASC specks released in the culture supernatant after HIV-1 infection, suggesting that macrophages remain viable after infection. Using in silico prediction studies, we found that the HIV-1 proteins Gag and Vpr interact with several host proteins. Comparable levels of trans-LTB4 were found in the supernatants of uninfected and HIV-1-infected macrophages, whereas ROS production was impaired in infected cells, which was not reversed after the PMA stimulus. Immunofluorescence analysis showed structural alterations in the mitochondrial architecture and an increase of BIM in the cytoplasm of infected cells. Our data suggest that HIV-1 proteins Gag and Vpr, through interacting with cellular proteins in the early steps of infection, preclude the inflammasome activation and the development of effective immune responses, thus allowing the establishment of the infection.


Increase in intracellular and extracellular myocardial mass in patients with acromegaly: a cardiac magnetic resonance imaging study.
Acromegaly is associated with an increased left ventricular (LV) mass, as reported in echo-based and, more recently, in a few cardiac magnetic resonance imaging (MRI) studies. One possible explanation for this increased LV mass could be water retention and subsequent myocardial edema. In this prospective cross-sectional study, 26 patients with active acromegaly before and after treatment and 31 controls of comparable age and sex were investigated using cardiac MRI. Cardiac morphology, function, and myocardial tissue characteristics were evaluated. Myocardial T2 relaxation time was used as the main outcome measure of myocardial edema. The study was registered with clinicaltrials.gov (NCT02948322). Patients compared to controls had greater LV mass indexes (58.1 [54.7-68.6] vs 46.0 [41.3-49.8] g/m2; P < .001) and end-diastolic volume (EDV) indexes (97.3 [88-101.2] vs 81.6 [78.1-96.2] mL/m2; P = .0069) and had comparable global contractile function. T2 values were not different between patients and controls. Both intracellular (43.83 [41.0-50.0] vs 34.32 [28.9-38.7] g/m2; P < .001) and extracellular (15.06 [13.5-17.1] vs 11.6 [10.8-12.7] g/m2; P < .001) LV mass indexes were higher in patients compared to controls. Log growth hormone correlated with myocardial mass (r = 0.75; P < .001). Sex, systolic blood pressure (BP), and the presence of acromegaly were predictors of the LV mass index. The extracellular LV mass index was associated with sex and the presence of acromegaly, whereas the intracellular LV mass index was associated with sex, systolic BP, and high-density lipoprotein (HDL) cholesterol. Acromegaly treatment reduced EDV and total and intracellular LV mass indexes without significantly affecting extracellular mass. Acromegaly results in a disease-specific form of LV hypertrophic remodeling, characterized by an increase in both intra- and extracellular mass. The LV mass index and intracellular mass were decreased by treatment.


Exploring the vertical transport of microplastics in subsurface environments: Lab-scale experiments and field evidence.
Microplastics (MPs) defined as smaller 5 mm plastic particles have received increasing attention due to their global occurrence and potential toxicity. This study investigated the effects of environmental factors (rainfall intensity, 13 and 29 mm/h) and MP characteristics (morphology (fiber, flake, and film), polymer type (polypropylene (PP), polyethylene terephthalate (PET), and polystyrene (PS)) and size (100-300, 300-500, and 500-1000 μm)) on the vertical transport of MP in unsaturated soil conditions using lab-scale column experiments. Additionally, the occurrence and characteristics of MP detected in soil/sediment (total 13 samples) and groundwater samples (total 6 samples) were explored in the field study. Laboratory-scale column experiments revealed that heavy rainfall intensity (29 mm/h) increased the degree of MP vertical transport in unsaturated soil conditions and MP fibers showed the greatest vertical mobility among the various morphologies of MPs assessed. For the polymer type and size, the lighter PP polymer or the larger size of MP (500-1000 μm) showed higher mobility. In the field study, a statistical difference in MP abundance was observed depending on the population density and degree of urban development in both soil and groundwater samples. Comparing to the two different types of environmental media samples obtained from the same site, there was a significant difference in the composition of polymer types present while statistically no difference in MP abundance was observed between the two media samples (i.e., soil or sediment and groundwater). In addition, MP fibers and polyethylene (PE) were predominantly detected in our two study areas. These results suggest that various types of MP can pass through the unsaturated zone by water infiltration, even if it takes a long time to reach groundwater. Overall, we found that the degree of vertical transport of the MPs was highly sensitive to environmental conditions and MP characteristics.


How do auditory cortex neurons represent communication sounds?
A major goal in auditory neuroscience is to characterize how communication sounds are represented at the cortical level. The present review aims at investigating the role of auditory cortex in the processing of speech, bird songs and other vocalizations, which all are spectrally and temporally highly structured sounds. Whereas earlier studies have simply looked for neurons exhibiting higher firing rates to particular conspecific vocalizations over their modified, artificially synthesized versions, more recent studies determined the coding capacity of temporal spike patterns, which are prominent in primary and non-primary areas (and also in non-auditory cortical areas). In several cases, this information seems to be correlated with the behavioral performance of human or animal subjects, suggesting that spike-timing based coding strategies might set the foundations of our perceptive abilities. Also, it is now clear that the responses of auditory cortex neurons are highly nonlinear and that their responses to natural stimuli cannot be predicted from their responses to artificial stimuli such as moving ripples and broadband noises. Since auditory cortex neurons cannot follow rapid fluctuations of the vocalizations envelope, they only respond at specific time points during communication sounds, which can serve as temporal markers for integrating the temporal and spectral processing taking place at subcortical relays. Thus, the temporal sparse code of auditory cortex neurons can be considered as a first step for generating high level representations of communication sounds independent of the acoustic characteristic of these sounds. This article is part of a Special Issue entitled "Communication Sounds and the Brain: New Directions and Perspectives".


Considerations of Left Ventricular Assist Device Therapy in the United States' Prison System.
Left ventricular assist device therapy for advanced heart failure is contraindicated if a patient lives in an unsafe environment and recent guidelines declare that "legal history is pertinent for determining personal constraints or financial responsibilities due to parole requirements, pending charges, and possible imprisonment," implying that incarceration would be a contraindication. International guidelines and precedent in the United States establish that medical care for incarcerated persons should match access in the community. We present a case example and practical considerations for advanced heart failure programs faced with the challenge of partnering with patients with heart failure who may be incarcerated and their correctional health systems in the care of their chronic condition. We encourage the heart failure community to not let incarceration be a contraindication to left ventricular assist device therapy.


Galantamine augmentation of long-acting injectable risperidone for cognitive impairments in chronic schizophrenia.
Galantamine, a reversible cholinesterase inhibitor with effects on nicotinic receptors, has shown mixed effects on cognitive impairments in patients with schizophrenia. Given these mixed results we examined whether galantamine compared to adjunctive placebo may improve cognitive functions in patients treated concomitantly with a long acting atypical antipsychotic. The parent study was a 52-week double-blind, randomized study of treatment with long-acting injectable risperidone 25mg or 50mg every two weeks. Adjunctive galantamine or placebo treatment was administered from Month 6 to 12. Outcome measures were neurocognitive, psychopathology, social and quality of life functions. Patients were randomized to blinded galantamine up to 24mg/day or matching placebo tablets. All patients were maintained on their randomized long-acting injectable risperidone regimen for the duration of the trial. 32 patients were included in the intent-to-treat analysis. No statistically significant differences were found for Attention Vigilance, Declarative Memory, Processing Speed, Reasoning/Problem Solving, Working Memory domains and the Neurocognitive Composite Score. Group specific analysis showed a statistically significant group interaction (p=0.043) with the Social Cognition domain showing in the galantamine group significantly lower scores at endpoint than placebo patients. The PANSS general psychopathology subscale showed significantly higher scores in the galantamine group at endpoint (p=0.05). ANCOVA model for within treatment group comparisons showed a significant increase of 7.3 points for the total PANSS score for the galantamine group. Galantamine showed no ameliorative effects on cognitive measures in this 6month, double-blind study of patients with schizophrenia treated with an assured and stable antipsychotic medication delivery system. Galantamine may not be an appropriate augmentation agent for cognitive impairments in patients with schizophrenia at the dose used.


The Ror receptor tyrosine kinase family.
Receptor tyrosine kinases (RTKs) participate in numerous developmental decisions. Ror RTKs are a family of orphan receptors that are related to muscle specific kinase (MuSK) and Trk neurotrophin receptors. MuSK assembles acetylcholine receptors at the neuromuscular junction, and Trk receptors function in the developing nervous system (reviewed in [3-5]). Rors have been identified in nematodes, insects and mammals. Recent studies have begun to shed light on Ror function during development. In most species, Rors are expressed in many tissue types during development. Analyses of mutants that are defective in the single nematode Ror demonstrate a role in cell migration and in orienting cell polarity. Mice lacking one of the two Ror gene products display defects in bone and heart formation. Similarly, two different human bone development disorders, dominant brachydactyly B and recessive Robinow syndrome, result from mutations in one of the human Ror genes.


Impact of cancer metabolism on therapy resistance - Clinical implications.
Despite an increasing arsenal of anticancer therapies, many patients continue to have poor outcomes due to the therapeutic failures and tumor relapses. Indeed, the clinical efficacy of anticancer therapies is markedly limited by intrinsic and/or acquired resistance mechanisms that can occur in any tumor type and with any treatment. Thus, there is an urgent clinical need to implement fundamental changes in the tumor treatment paradigm by the development of new experimental strategies that can help to predict the occurrence of clinical drug resistance and to identify alternative therapeutic options. Apart from mutation-driven resistance mechanisms, tumor microenvironment (TME) conditions generate an intratumoral phenotypic heterogeneity that supports disease progression and dismal outcomes. Tumor cell metabolism is a prototypical example of dynamic, heterogeneous, and adaptive phenotypic trait, resulting from the combination of intrinsic [(epi)genetic changes, tissue of origin and differentiation dependency] and extrinsic (oxygen and nutrient availability, metabolic interactions within the TME) factors, enabling cancer cells to survive, metastasize and develop resistance to anticancer therapies. In this review, we summarize the current knowledge regarding metabolism-based mechanisms conferring adaptive resistance to chemo-, radio-and immunotherapies as well as targeted therapies. Furthermore, we report the role of TME-mediated intratumoral metabolic heterogeneity in therapy resistance and how adaptations in amino acid, glucose, and lipid metabolism support the growth of therapy-resistant cancers and/or cellular subpopulations. We also report the intricate interplay between tumor signaling and metabolic pathways in cancer cells and discuss how manipulating key metabolic enzymes and/or providing dietary changes may help to eradicate relapse-sustaining cancer cells. Finally, in the current era of personalized medicine, we describe the strategies that may be applied to implement metabolic profiling for tumor imaging, biomarker identification, selection of tailored treatments and monitoring therapy response during the clinical management of cancer patients.


Chief veterinary officer's update: Big data is our best shot at challenging extreme breeding.
In his latest update for Veterinary Record, Nigel Gibbens, the UK's Chief Veterinary Officer, looks back at the past year and discusses the UK's achievements and the challenges that it faces.


Nitric oxide in invertebrates.
Nitric oxide (NO) is considered an important signaling molecule implied in different physiological processes, including nervous transmission, vascular regulation, immune defense, and in the pathogenesis of several diseases. The presence of NO is well demonstrated in all vertebrates. The recent data on the presence and roles of NO in the main invertebrate groups are reviewed here, showing the widespread diffusion of this signaling molecule throughout the animal kingdom, from higher invertebrates down to coelenterates and even to prokaryotic cells. In invertebrates, the main functional roles described for mammals have been demonstrated, whereas experimental evidence suggests the presence of new NOS isoforms different from those known for higher organisms. Noteworthy is the early appearance of NO throughout evolution and striking is the role played by the nitrergic pathway in the sensorial functions, from coelenterates up to mammals, mainly in olfactory-like systems. All literature data here reported suggest that future research on the biological roles of early signaling molecules in lower living forms could be important for the understanding of the nervous-system evolution.


Effect of a combination DNA vaccine for the prevention and therapy of Trypanosoma cruzi infection in mice: role of CD4+ and CD8+ T cells.
Chagas disease is a major public health problem, with about 10 million infected people, and DNA vaccines are a promising alternative for the control of Trypanosoma cruzi, the causing agent of the disease. We tested here a new DNA vaccine encoding a combination of two leading parasite antigens, TSA-1 and Tc24, for the prevention and therapy of T. cruzi infection. Immunized Balb/c mice challenged by T. cruzi presented a significantly lower parasitemia and inflammatory cell density in the heart compared to control mice. Similarly, the therapeutic administration of the DNA vaccine was able to significantly reduce the parasitemia and inflammatory reaction in acutely infected Balb/c and C57BL/6 mice, and reduced cardiac tissue inflammation in chronically infected ICR mice. Therapeutic vaccination induced a marked increase in parasite-specific IFNγ producing CD4(+) and CD8(+) T cells in the spleen as well as an increase in CD4(+) and CD8(+) T cells in the infected cardiac tissue. In addition, some effect of the DNA vaccine could still be observed in CD4-knockout C57BL/6 mice, which presented a lower parasitemia and inflammatory cell density, but not in CD8-deficient mice, in which the vaccine had no effect. These results indicate that the activation of CD8(+) T cells plays a major role in the control of the infection by the therapeutic DNA vaccine, and to a somewhat lesser extent CD4(+) T cells. This observation opens interesting perspectives for the potentiation of this DNA vaccine candidate by including additional CD8(+) T cell antigens/epitopes in future vaccine formulations.


A new mini-transposon for in vivo protein epitope tagging: application to Burkholderia multivorans.
A short amino acid sequence coding for the mature Pseudomonas aeruginosa OprI lipoprotein was fused to a mini-Tn5 plasposon (mini-transposon with an origin of replication) with tetracycline resistance in order to generate in-frame fusion proteins after transposition. After conjugative transfer to Burkholderia multivorans, clones reacting with an anti-OprI mab were selected. In-frame OprI-tagged proteins were detected and identified for six clones. The six C-tagged proteins were detected by immunoblot. The different mutants had insertions into a histone H1-like coding gene, cspD, encoding a cold-shock protein, dsbC, encoding a putative outer membrane lipoprotein involved in thiol-disulfide exchange, paaE, a ferredoxin-NADPH reductase gene, a gene for the catabolism of propionate, and one encoding an unknown protein.


A geometric morphometric evaluation of hard and soft tissue profile changes in borderline extraction versus non-extraction patients.
To evaluate the hard tissue and facial profile changes in matched extraction and non-extraction Class I patients by the use of geometric morphometrics. From a parent sample of 542 Class I patients, previously subjected to discriminant analysis, a subsample of 68 borderline cases was obtained, 34 treated with extraction and 34 without extraction of 4 first premolars. Geometric morphometric methods (Procrustes superimposition and Principal Component Analysis) were applied on cephalometric tracings to assess the validity of the discriminant analysis in successfully identifying a morphologically homogeneous group and to evaluate inter- and intra-group skeletal and facial profile shape changes. No significant pre-treatment shape difference between the two groups was found, thus validating the discriminant analysis. The non-extraction group showed increase in hard tissue facial height (P < 0.001), with slight lower lip retrusion and upper lip protrusion (P = 0.027). The extraction group showed retraction of the hard tissue and facial profile outline (P < 0.001). Permutation tests for post-treatment inter-group differences resulted in P = 0.054 for the soft tissue outline and P = 0.078 for the hard tissue skeletal component. The evidence indicates that borderline cases treated with four premolar extractions will exhibit lip retrusion compared to non-extraction treatment.


Progression rate of myelopathy in X-linked adrenoleukodystrophy heterozygotes.
X-linked adrenoleukodystrophy heterozygote women can present adult onset myeloneuropathy and little is known about its natural history. We aimed to describe the progression rate of the neurological impairment in the prospective follow-up of our cohort and to look for prognostic factors. The neurological scales Japanese Orthopaedic Association (JOA) and Severity Score System for Progressive Myelopathy (SSPROM) were applied at baseline in 29 symptomatic carriers and in follow-up visits. Age at onset, disease duration, X inactivation pattern, determination of the allele expressed, plasma levels of the very long chain fatty acids and of the neuron-specific enolase, and somato-sensory evoked potentials, were taken at baseline. The slope of the linear regression of both JOA and SSPROM versus disease duration since the first symptom was estimated using mixed modeling. JOA and SSPROM decreased 0.42 and 1.87 points per year, respectively (p < 0.001). None of the parameters under study influenced these rates. We estimated that the number of carriers per arm needed in a future 12 month trial with 80% power and a 50% reduction in disease progression would be 225 women for JOA and 750 for SSPROM. The progression rates of the studied neurological scales were small, did not depend on any modifier factor known, and reflected the characteristically slow worsening of symptoms in X-ALD heterozygotes. Better biomarkers are still necessary for future studies.


Thermo diffusion impacts on the flow of second grade fluid with application of (ABC), (CF) and (CPC) subject to exponential heating.
The aim of this article is to investigate the exact solution by using a new approach for the thermal transport phenomena of second grade fluid flow under the impact of MHD along with exponential heating as well as Darcy's law. The phenomenon has been expressed in terms of partial differential equations, then transformed the governing equations in non-dimentional form. For the sake of better rheology of second grade fluid, developed a fractional model by applying the new definition of Constant Proportional-Caputo hybrid derivative (CPC), Atangana Baleanu in Caputo sense (ABC) and Caputo Fabrizio (CF) fractional derivative operators that describe the generalized memory effects. For seeking exact solutions in terms of Mittag-Leffler and G-functions for velocity, temperature and concentration equations, Laplace integral transformation technique is applied. For physical significance of various system parameters on fluid velocity, concentration and temperature distributions are demonstrated through various graphs by using graphical software. Furthermore, for being validated the acquired solutions, accomplished a comparative analysis with some published work. It is also analyzed that for exponential heating and non-uniform velocity conditions, the CPC fractional operator is the finest fractional model to describe the memory effect of velocity, energy and concentration profile. Moreover, the graphical representations of the analytical solutions illustrated the main results of the present work. Also, in the literature, it is observed that to derived analytical results from fractional fluid models developed by the various fractional operators, is difficult and this article contributing to answer the open problem of obtaining analytical solutions the fractionalized fluid models.


Characterisation of CadR from Pseudomonas aeruginosa: a Cd(II)-responsive MerR homologue.
cadR from Pseudomonas aeruginosa encodes a transcriptional regulatory protein which responds to Cd(II)>>Zn(II)>Hg(II) at its cognate promoter PcadA. CadR will also act to induce transcription at the Escherichia coli ZntR cognate promoter, PzntA, however, the induction profile is altered to Hg(II)>Cd(II)>Zn(II). Two separate single base pair deletions within PzntA result in further alteration of relative specificity in metal-ion induction profile for CadR. This demonstrates that the operator/promoter sequence can play a role in defining optimal ligand response and that for these regulators specificity is not solely a function of the regulatory protein.


Pharmacokinetics of dexmedetomidine combined with therapeutic hypothermia in a piglet asphyxia model.
The highly selective α2 -adrenoreceptor agonist, dexmedetomidine, exerts neuroprotective, analgesic, anti-inflammatory and sympatholytic properties that may be beneficial for perinatal asphyxia. The optimal safe dose for pre-clinical newborn neuroprotection studies is unknown. Following cerebral hypoxia-ischaemia, dexmedetomidine was administered to nine newborn piglets in a de-escalation dose study in combination with hypothermia (whole body cooling to 33.5°C). Dexmedetomidine was administered with a loading dose of 1 μg/kg and maintenance infusion at doses from 10 to 0.6 μg/kg/h. One additional piglet was not subjected to hypoxia-ischaemia. Blood for pharmacokinetic analysis was sampled pre-insult and frequently post-insult. A one-compartment linear disposition model was used to fit data. Population parameter estimates were obtained using non-linear mixed effects modelling. All dexmedetomidine infusion regimens led to plasma concentrations above those associated with sedation in neonates and children (0.4-0.8 μg/l). Seven out of the nine piglets with hypoxia-ischaemia experienced periods of bradycardia, hypotension, hypertension and cardiac arrest; all haemodynamic adverse events occurred in piglets with plasma concentrations greater than 1 μg/l. Dexmedetomidine clearance was 0.126 l/kg/h [coefficient of variation (CV) 46.6.%] and volume of distribution was 3.37 l/kg (CV 191%). Dexmedetomidine clearance was reduced by 32.7% at a temperature of 33.5°C. Dexmedetomidine clearance was reduced by 55.8% following hypoxia-ischaemia. Dexmedetomidine clearance was reduced almost tenfold compared with adult values in the newborn piglet following hypoxic-ischaemic brain injury and subsequent therapeutic hypothermia. Reduced clearance was related to cumulative effects of both hypothermia and exposure to hypoxia. High plasma levels of dexmedetomidine were associated with major cardiovascular complications.


Genetic restriction of protective immunity to Plasmodium yoelii sporozoites.
Ten congenic strains of mice were immunized with irradiated sporozoites of Plasmodium yoelii. When challenged with viable sporozoites, only two strains had a high proportion of animals which did not develop blood-stage infections. Genes both within and outside the H-2 region affected the degree of protection. Immunity did not correlate with anti-sporozoite antibody levels. In vivo depletion of CD8+ T cells did not alter immunity in two of three congenic strains, implying the existence of a novel mechanism of cell-mediated immunity.


Transanal endoscopic microsurgery: a Brazilian initial experience in private practice.
Transanal endoscopic microsurgery is a minimally invasive approach for rectal lesions. Superior exposure and access to the entire rectum result in lesser risk of compromised margin and lower recurrence rates compared to conventional transanal excision. It was aimed at describing a single institution's initial experience with transanal endoscopic microsurgery. Retrospective review of a prospective database. Fifty-two procedures from March 2009 to November 2011 were analyzed. Fifty operations were completed. There were 23 men. Mean age was 67.5 (42-89). Mean follow-up was 23 (1-31) months. Average tumor size was 4.8 cm (1.5-14 cm). Mean distance from anal verge was 6 (3-15) cm. Mean operating time was 110 (86-170) min. Postoperative complication rate was 16%o. There were no re-admissions. Mortality was null. Operative pathology was adenoma in 25, in situ adenocarcinoma in eight, invasive adenocarcinoma in 13, neuroendocrine carcinoma in three and no residual lesion in one case. Recurrence was 4% for benign and 8% for malignant tumors. TEM is a minimally invasive procedure with low postoperative morbidity during initial experience. TEM is curative for benign lesions and for selected early cancers. It is useful after neoadjuvant therapy for strictly selected cancers while the results of multi-institutional trials are awaited.


Determination of the quantitative stereoselectivity fingerprint of lipases during hydrolysis of a prochiral triacylglycerol.
We propose a method for characterizing quantitatively the stereoselectivity of lipases during hydrolysis of triacylglycerols. Although it is of general applicability, we demonstrate it specifically for sn-1,3-regiospecific lipases. In this case the method generates a "stereoselectivity fingerprint" that consists of ratios of the specificity constants for the various reactions that produce and consume the 1,2-sn- and 2,3-sn-diacylglycerols. We use the method to determine the stereoselectivity fingerprint of several lipases during the hydrolysis of the prochiral substrate triolein. Our method opens up the possibility of correlating quantitative fingerprints with structural information, in the quest to elucidate the mechanisms underlying the stereoselectivity of lipases.


Simple and Fast Assay for Apolipoprotein E Phenotyping and Glycotyping: Discovering Isoform-Specific Glycosylation in Plasma and Cerebrospinal Fluid.
The mechanisms of how APOEɛ4 allele (APOE4) increases the risk of Alzheimer's disease (AD) pathology have not been fully elucidated. In cerebrospinal fluid (CSF), apoE is heavily glycosylated. To determine the impact of APOE genotype on the relative abundance of apoE protein isoforms and their specific glycosylation patterns in CSF and plasma via a newly developed mass spectrometric immunoassay (MSIA) assay. Total glycosylation and isoform-specific glycosylation were analyzed in plasma and CSF from a group of non-demented older individuals (n = 22), consisting of homozygous ɛ3 and ɛ4 or heterozygous ɛ3/ɛ4, ɛ2/ɛ3, or ɛ2/ɛ4 carriers. The glycan structures were further confirmed after treatment with sialidase. In heterozygous individuals, the apoE3/E2, E4/E2, and E4/E3 isoform ratios were all significantly lower in plasma compared to CSF. For all individuals, a single O-linked glycan was observed in plasma, while two glycans (of the same type) per apoE were observed in CSF. The ratio of glycosylated to total apoE was greater in CSF compared to plasma for all apoE isoforms. In plasma and CSF, a trend of decreasing glycosylation was observed from apoE2 > apoE3 > apoE4. The difference in the percentage of secondary glycosylation in CSF was significantly greater in apoE4 compared to the other isoforms. The new MSIA apoE assay robustly distinguishes among apoE isoforms and glycoforms in plasma and CSF. ApoE4 is the predominant isoform and least glycosylated in CSF. Assessing apoE isoform-specific glycosylation by MSIA may help clarify brain apoE metabolism and AD risk.


Accuracy of predictive equations for resting energy expenditure estimation in mechanically ventilated Thai patients.
Indirect calorimetry (IC) is the most precise approach for estimating calorie demand in critically ill patients. Despite this, owing to unaffordable devices, it is rarely used in practice. Predictive equations are the alternatives. To assess the accuracy of 14 predictive resting energy expenditure(REE) equations in ventilated Thai patients. We compared the accuracy and agreement of 14 equations. The equations included the American College of Chest Physicians(ACCP) equation, Harris-Benedict equation(HBE), 1.2×HBE, 1.5×HBE, Mifflin-St. Jeor(MSJ), Ireton-Jones 1992 and 2002, Penn State 2003(HBE and MSJ) and 2010, Swinamer 1990, Faisy, Brandi 1999, and 25 kcal/kg equation. An equation was ascertained as accurate if the calculated values fell within ±10% of the measured REEs. Spearman correlation coefficient, Bland-Altman method, and intraclass correlation coefficient were used to analysis. We obtained data from 24 ventilated patients undergoing REE measurement by IC. Fifty percent of them were male with a median age of 64.5 years, a median height of 160 cm, and a median body mass index of 22.95 kg/m2. The predictive precision of all equations was poor, with largely different accuracies from 6.7% to 48.1%. The most reliable equation was Penn State 2010. The ACCP, HBE, MSJ, and Penn State 2003(HBE) tended to underestimate calorie need. Contrastingly, the other equations tended to overestimate REEs. Despite a moderate degree of correlations, the Bland-Altman plots demonstrated clinically unacceptable discrepancies between measured REE and REE calculated by each equation. In ventilated Thai patients, there were no precise equations for determining REE.


Life stress and psychopathology: progress on research begun with Barbara Snell Dohrenwend.
This is a progress report on some of the research that was planned and begun with Barbara Dohrenwend before she died in 1982. The main focus is on two of the studies. One was conducted in New York City; the other is still underway in Israel. The New York study is a retrospective case/control study of social and psychological factors that may put people at risk for developing schizophrenic episodes and episodes of major depression. The Israel research consists of epidemiological, case/control, and family studies of these two disorders together with other types of psychopathology that are inversely related to social class. Preliminary findings from both studies are reported, and their implications for primary prevention are discussed.


Position of the American Dietetic Association: the impact of fluoride on health.
The American Dietetic Association reaffirms that fluoride is an important element for all mineralized tissues in the body. Appropriate fluoride exposure and usage is beneficial to bone and tooth integrity and, as such, has an important, positive impact on oral health as well as general health throughout life. Fluoride is an important element in the mineralization of bone and teeth. The proper use of topical and systemic fluoride has resulted in major reductions in dental caries (tooth decay) and its associated disability. The Centers for Disease Control and Prevention have named fluoridation of water as one of the 10 most important public health measures of the 20th century. Nearly 100 national and international organizations recognize the public health benefits of community water fluoridation for preventing dental caries. However, by the year 2000, over one third of the US population (over 100 million people) were still without this critical public health measure. Fluoride also plays a role in bone health. However, the use of high doses of fluoride for prevention of osteoporosis is considered experimental at this point. Dietetics professionals should routinely monitor and promote the use of systemic and topical fluorides, especially in children and adolescents. The American Dietetic Association strongly reaffirms its endorsement of the appropriate use of systemic and topical fluorides, including water fluoridation, at appropriate levels as an important public health measure throughout the life span.


EBV-transformed lymphoblastoid cell lines as vaccines against cancer testis antigen-positive tumors.
EBV-transformed lymphoblastoid cell lines (LCL) are potent antigen-presenting cells. To investigate their potential use as cancer testis antigen (CTA) vaccines, we studied the expression of 12 cancer testis (CT) genes in 20 LCL by RT-PCR. The most frequently expressed CT genes were SSX4 (50 %), followed by GAGE (45 %), SSX1 (40 %), MAGE-A3 and SSX2 (25 %), SCP1, HOM-TES-85, MAGE-C1, and MAGE-C2 (15 %). NY-ESO-1 and MAGE-A4 were found in 1/20 LCL and BORIS was not detected at all. Fifteen of 20 LCL expressed at least one antigen, 9 LCL expressed ≥2 CT genes, and 7 of the 20 LCL expressed ≥4 CT genes. The expression of CT genes did not correlate with the length of in vitro culture, telomerase activity, aneuploidy, or proliferation state. While spontaneous expression of CT genes determined by real-time PCR and Western blot was rather weak in most LCL, treatment with DNA methyltransferase 1 inhibitor alone or in combination with histone deacetylase inhibitors increased CTA expression considerably thus enabling LCL to induce CTA-specific T cell responses. The stability of the CT gene expression over prolonged culture periods makes LCL attractive candidates for CT vaccines both in hematological neoplasias and solid tumors.


Paraoxonase single nucleotide variants show associations with polycystic ovary syndrome: a meta-analysis.
Etiology of polycystic ovary syndrome (PCOS) is attributed to genetic and environmental factors. One environmental factor is oxidative stress. Paraoxonase 1 (PON1) is an antioxidant high-density lipoprotein-associated enzyme encoded by the PON1 gene. The PON1 gene has been implicated in the risk for PCOS, the influence of which appears to come from single nucleotide variants (SNVs) at multiple genetic loci. However, association study reports have been inconsistent which compels a meta-analysis to obtain more precise estimates. From 12 publications, extracted genotype data were used in two genetic procedures. First, linkage disequilibrium (LD) was used to group eight PON SNVs into three: LD1, LD2 and LD3. Second, frequencies of the variant (var), wild-type (wt) and heterozygous (het) genotypes were used for genetic modeling (allele-genotype for LD1 and standard for LD2 and LD3). Risk associations were expressed in terms of pooled odds ratios (ORs), 95% confidence intervals (CIs) and Pa-values. Evidence was considered strong when significance was high (Pa < 0.0001) and heterogeneity absent (I2 = 0%). Pooled effects were subjected to modifier (power), subgroup (Asian/Caucasian), outlier, sensitivity and publication bias treatments. Multiple comparisons were Bonferroni-corrected. This meta-analysis generated 11 significant outcomes, five in LD1, six in LD2 and none in LD3. All six LD2 outcomes did not survive the Bonferroni-correction but two of the five in LD1 did. These two core LD1 findings conferred greater odds of PCOS to the var allele in the highly significant (Pa < 0.0001) overall (OR 1.44, 95% CI 1.24-1.67) and Asian (OR 1.41, 95% CI 1.20-1.65) outcomes. Of these two core outcomes, the Asian effect was homogeneous (I2 = 0%) but not the overall (I2 = 29%). Of the eight PON SNVs examined, two (rs854560 and rs662) were associated with PCOS risk. These 1.4-fold increased risk effects rendered Asians susceptible to PCOS. High statistical power, high significance, zero to low-level heterogeneity, robustness and lack of bias in the core outcomes underpinned the strong evidence for association.


Discovery of Covalent MKK4/7 Dual Inhibitor.
MKK4/7 are kinases that phosphorylate JNKs and regulate the MAPK signaling pathway. Their overexpression has been associated with tumorigenesis and aggressiveness in cancers such as breast, prostate, non-small cell lung, and pediatric leukemia, making them a potential target for inhibitor development. Here, we report the discovery, development, and validation of a dual MKK4/7 inhibitor, BSJ-04-122, that covalently targets a conserved cysteine located before the DFG motif and displays excellent kinome selectivity. BSJ-04-122 exhibits potent cellular target engagement and induces robust target-specific downstream effects. The combination of the dual MKK4/7 inhibitor with a selective, covalent JNK inhibitor demonstrated an enhanced antiproliferative activity against triple-negative breast cancer cells. Taken together, the results show that BSJ-04-122 represents a pharmacological probe for MKK4/7 and credential covalent targeting as a way to explore the therapeutic potential of these kinases.


An endogenous substrate for the insulin receptor-associated tyrosine kinase.
Insulin binding to its receptor stimulates a tyrosine-specific protein kinase. This enzyme phosphorylates the insulin receptor, as well as a variety of exogenous substrates in vitro. In the present studies, we have identified an endogenous substrate for the insulin receptor-associated kinase. We studied insulin-stimulated protein phosphorylation in partially purified insulin receptor preparations from the livers of dexamethasone-treated rats. In this cell-free system, insulin stimulated the phosphorylation of its own receptor as well as of a phosphoprotein of apparent Mr = 120,000 (termed pp120). pp120 was not immunoprecipitated by three anti-receptor antisera, nor was the receptor immunoprecipitated by antisera raised against pp120, suggesting that pp120 is not antigenically related or tightly bound to the insulin receptor. Dose-response curves for receptor and pp120 phosphorylation stimulated by pork insulin were essentially identical, and showed the appropriate specificity (insulin much greater than proinsulin) for a receptor-mediated event. Phosphoamino acid analysis revealed that insulin stimulated the incorporation of 32P predominantly into tyrosine residues of pp120. Casein, an artificial substrate for the insulin receptor kinase, competed with pp120 for insulin-stimulated phosphorylation. Phosphorylation of pp120 was rapid (half-maximal effect within 2 min at 24 degrees C) and, like receptor phosphorylation, was supported with Mn2+ or Mg2+ as divalent cation and ATP as the phosphate donor. While receptor autophosphorylation and artificial substrate phosphorylation were not altered by prior treatment of the rats with dexamethasone, insulin-stimulated pp120 phosphorylation was enhanced in preparations derived from dexamethasone-treated rats, suggesting an alteration of pp120, not the receptor, as a result of dexamethasone-treatment. Further studies of this newly identified endogenous substrate may help clarify the physiologic role of the insulin receptor-associated kinase.


Delayed enterocutaneous fistula after 17 years of gunshot wound injury: a case report.
Enterocutaneous fistula (ECF) is a common complication of abdominal surgery. Postoperatively, it may be the result of a small intestinal injury or an anastomosis leak, and it typically manifests days or weeks following surgery. A 56-year-old man with a history of a gunshot wound injury in the abdomen with colon injury managed for colocolic anastomosis 17 years earlier presented to our hospital's general surgery clinic with the complaint of drainage at the scar area. It was discovered that he had an ECF. The patient underwent a laparotomy. Fistula tract excision and segmental colon resection with colocolic anastomosis were done. ECF formation after an extraordinarily long latency due to an anastomotic leak, which has not been previously documented in the literature, is a distinctive aspect of the case. As a result, surgeons should consider ECF in their differentials for patients with a history of abdominal operations. Surgical management is the definitive treatment of ECF after proper resuscitation and stabilization of the patient's condition. Preoperative imaging is crucial for determining the anatomy of the fistula and any associated intra-abdominal pathology.


Calcium release and intramembranous charge movement in frog skeletal muscle fibres with reduced (< 250 microM) calcium content.
It is generally accepted that activation of voltage sensors in the T-tubular membranes is a critical step of excitation-contraction coupling in skeletal muscle. The purpose of this study was to evaluate further whether the Qgamma component (delayed 'hump' component) of the intramembranous charge movement current (I(cm)) results from movement of these voltage sensors. Ca2+ release and I(cm) were measured in voltage-clamped frog cut fibres mounted in a double Vaseline-gap chamber. In order to reduce effects of Ca2+ feedback mechanisms, the calcium content of the sarcoplasmic reticulum (SR) during rest was reduced to < 250 microM (referred to volume of myoplasm) and maintained approximately constant. The early (Qbeta) and Qgamma components of charge movement were estimated by fitting the sum of two Boltzmann functions to the total steady-state intramembranous charge vs. voltage data. The average voltage steepness factor (k) and half-maximal voltage (V-) for Qgamma were 4.3 and -57.4 mV (n = 6), respectively. The SR membrane permeability for Ca2+ release was assessed when a constant amount of calcium remained in the SR (usually about 60 microM). A single Boltzmann function fitted to these data gave values on average for k and V- of 4.7 and -45.3 mV, respectively. The similarity of the values of k for Qgamma and Ca2+ release supports the idea that Qgamma reflects movement of voltage sensors for Ca2+ release. The greater value of V- for Ca2+ release compared to Qgamma is consistent with multi-state models of the voltage sensor involving movement of Qgamma charge during non-activating transitions.


Degradation of peptides and proteins of different sizes by homogenates of human MRC5 lung fibroblasts. Aged cells have a decreased ability to degrade shortened proteins.
The degradation of haemoglobin and haemoglobin-derived peptide fragments by homogenates of MRC5 fibroblasts has been investigated. Results show that the smaller fragments were degraded more rapidly than larger substrates at both pH 5.5 and pH 7.5. Only the smallest of the soluble cyanogen bromide peptides (Mr 3500) was degraded at pH 7.5. Degradation at pH 5.5 proceeded more rapidly than that at pH 7.5 for all substrates tested but was more marked with the larger substrates. Homogenates prepared from aged cells degraded puromycin peptides and, to a lesser extent, cyanogen bromide peptides at a slower rate, at pH 7.5, than those prepared from younger cells. We suggest that cytosolic degradation is less selective and at least one cytosolic proteolytic activity decreases as cells age.


[Successful treatment of a case of extense verrucous chromoblastomychosis with surgical exeresis associated with the use of ketoconazole].
The case of a 56-year-old man with an extense verrucous chromoblastomycosis of long evolution, with lesions in the right foot that had not been cured with previous treatment, and that was successfully treated by surgical exeresis of all the lesions together with oral ketoconazole, is presented. Literature was reviewed to conclude that the treatment of chromoblastomycosis is still a challenge and that it should be selected for each case in particular.


Influence of the Ring Size on the Binding Ability of FTO Investigated by Fluorescence Spectroscopy.
The fat mass and obesity associated protein (FTO) is a potential target for anti-obesity medicines. In this paper, we have synthesized two potential inhibitors for FTO, three-member-ring compound (W 3 ) and four-member-ring compound (W 4 ). The interactions of fat mass and obesity-associated (FTO) protein with W 3 (or W 4 ) have been studied by spectral method. Results show the intrinsic fluorescence is quenched by the W 3 (or W 4 ). The thermodynamics parameters indicate hydrophobic interaction play a major role in the interactions. The results of synchronous fluorescence spectra demonstrate that the microenvironments of Trp residue of FTO are disturbed by W 3 and W 4 . Results showed that W 3 are stronger quenchers and bind to FTO with the higher affinity than W 4 . The influence of molecular structure on the binding aspects has been investigated.


An alarming rise in the prevalence of deaths with methamphetamine involved in Tehran, Iran 2011-2018.
Methamphetamine is one of the most commonly abused illicit drugs in Iran. Despite this fact, there has been relatively little research on methamphetamine-associated deaths in Iran. The present study aimed to investigate the cause of death in methamphetamine positive cases based on forensic toxicology findings In this data base descriptive study, individuals with methamphetamine-associated deaths that were referred to the Legal Medicine Organization, Tehran, Iran (2011-2018) were assessed. Deaths including those from natural causes, toxicity, accident and suicides with positive forensic toxicology analyses for methamphetamine and its metabolite amphetamine in postmortem samples were investigated. All cases were examined to determine trends in methamphetamine-associated deaths, and in the manner and causes of deaths. During the eight year study there were 1389 methamphetamine-associated deaths. The mean age was 37.47±10.87 years, with a male/female ratio of 12.36:10. The methamphetamine-associated death rate increased significantly during the study period (2.05 vs. 21.93 per 106 Tehran province populations). The most common cause of death was cardiac arrest due to stimulant abuse with supportive histologic findings of acute myocardial infarction, myocardial fiber hypertrophy, aortic dissection and perivascular fibrosis (25%). Opioids (methadone, tramadol and morphine), tricyclic antidepressants and benzodiazepines were frequently present in postmortem samples of study cases. Methamphetamine-associated deaths significantly contribute to illicit drug-associated deaths in Tehran and represent a substantial clinical and public health problem.


First record of natural Tritrichomonas foetus infection of the feline uterus.
Tritrichomonas foetus was found in the uterus of a cat with pyometra and in the faeces of three other cats in the same household, one of which had chronic diarrhoea. This is the first report of a feline uterine infection with T. foetus and also the first time T. foetus has ever been diagnosed in animals in Norway. The diagnosis was made by microscopic examination and sequencing studies.


The Pap smear revisited.
Papanicolaou smear screening for cervical cancer has become an established practice in most developed countries. This is because the cervix is relatively accessible to investigation and treatment, and early stages in the morphogenesis of cervical cancer are both recognizable and easily treated. The Pap smear is a valid test. It is simple, relatively inexpensive, reliable, and free of risk. Although the test has far from perfect sensitivity, it has high specificity, and false-positive results are rare. In most reported series, the majority of false-negative results have been found to be attributable to collection errors rather than laboratory errors. Despite the importance of Pap smear screening, controlled prospective trials have not been undertaken to determine its efficiency in reducing cervical cancer incidence and mortality. However, countries with well-organized programmes, wide population coverage and correct follow-up appear to have had some impact on mortality from cervical cancer. Nevertheless, coverage of high-risk groups, particularly women over 40 years of age, remains the greatest problem. Recommendations on the frequency of testing vary considerably. Statistical models indicate triennial testing may deliver almost all of the effectiveness of annual testing at a substantially reduced cost, but the numerous reports of false-negative results argue strongly in favour of annual screening. It is possible that these problems may be solved in the future by increasing the sensitivity of the test and/or by the use of additional tests.


Genomic analysis of transcriptional networks directing progression of cell states during MGE development.
Homeodomain (HD) transcription factor (TF) NKX2-1 critical for the regional specification of the medial ganglionic eminence (MGE) as well as promoting the GABAergic and cholinergic neuron fates via the induction of TFs such as LHX6 and LHX8. NKX2-1 defines MGE regional identity in large part through transcriptional repression, while specification and maturation of GABAergic and cholinergic fates is mediated in part by transcriptional activation via TFs such as LHX6 and LHX8. Here we analyze the signaling and TF pathways, downstream of NKX2-1, required for GABAergic and cholinergic neuron fate maturation. Differential ChIP-seq analysis was used to identify regulatory elements (REs) where chromatin state was sensitive to change in the Nkx2-1cKO MGE at embryonic day (E) 13.5. TF motifs in the REs were identified using RSAT. CRISPR-mediated genome editing was used to generate enhancer knockouts. Differential gene expression in these knockouts was analyzed through RT-qPCR and in situ hybridization. Functional analysis of motifs within hs623 was analyzed via site directed mutagenesis and reporter assays in primary MGE cultures. We identified 4782 activating REs (aREs) and 6391 repressing REs (rREs) in the Nkx2-1 conditional knockout (Nkx2-1cKO) MGE. aREs are associated with basic-Helix-Loop-Helix (bHLH) TFs. Deletion of hs623, an intragenic Tcf12 aRE, caused a reduction of Tcf12 expression in the sub-ventricular zone (SVZ) and mantle zone (MZ) of the MGE. Mutation of LHX, SOX and octamers, within hs623, caused a reduction of hs623 activity in MGE primary cultures. Tcf12 expression in the SVZ of the MGE is mediated through aRE hs623. The activity of hs623 is dependent on LHX6, SOX and octamers. Thus, maintaining the expression of Tcf12 in the SVZ involves on TF pathways parallel and genetically downstream of NKX2-1.


Histaminergic activation of endogenous angiotensin II fails to inhibit alcohol intake in rats.
Demonstrations that alcohol intake can be inhibited by pharmacological activation of the renal renin-angiotensin system (RRA) or injection of angiotensin II (ANG II) in rats led to this study of a role for endogenous ANG II in inhibition of alcohol intake in rats. Relatively small doses of histamine, above threshold for eliciting drinking of water and activation of the RRA, were injected SC in adult male Sprague-Dawley rats with access to 3.0% alcohol in 45-min one-bottle alcohol tests and in two-bottle tests in which alcohol and water were available at the midpoint of the 12-h dark phase. The 0.312 and 1.25 mg/kg doses of SC histamine elevated plasma renin activity to levels similar to those in rats that had just eaten food. Neither dose of histamine affected alcohol intake in one-bottle tests. A relatively large 10 mg/kg dose of histamine increased alcohol intake in a one-bottle test, but decreased alcohol intake and increased water intake in two-bottle tests. The inhibitory effect of the 10 mg/kg dose of histamine on alcohol intake was completely blocked by SC 10 mg/kg losartan, a selective AT1 angiotensin receptor antagonist. This 10 mg/kg dose of losartan given alone, however, failed to increase alcohol intake in one- or two-bottle tests. These results generally do not support a role for endogenous ANG II as an inhibitory physiological signal in the control of alcohol ingestion in rats, because histaminergic activation of RRA, using small but physiologically meaningful doses of histamine, failed to inhibit alcohol intake.


Diabetes prevention at scale: Narrative review of findings and lessons from the DIPLOMA evaluation of the NHS Diabetes Prevention Programme in England.
The NHS Diabetes Prevention Programme (NHS DPP) is a large-scale, England-wide behaviour change programme for people at high risk of progressing to type 2 diabetes. We summarise the findings of our six-year DIPLOMA evaluation of its implementation and impact and highlight insights for future programmes. Using qualitative interviews, document analysis, observation, surveys and large dataset analysis, eight interlinked work packages considered: equity of access; implementation; service delivery and fidelity; programme outcomes; comparative effectiveness and cost-effectiveness in reducing diabetes incidence; and patient decision making and experience. Delivery of the NHS DPP encountered barriers across many aspects of the programme, and we identified inequalities in terms of the areas, organisations and patient populations most likely to engage with the programme. There was some loss of fidelity at all stages from commissioning to participant understanding. Despite these challenges, there was evidence of significant reductions in diabetes incidence at individual and population levels. The programme was cost-effective even within a short time period. Despite the challenge of translating research evidence into routine NHS delivery at scale, our findings suggest that an individual-level approach to the prevention of type 2 diabetes in a 'high-risk' population was more effective than usual care. By embedding evaluation with programme delivery and working closely with the NHS DPP team, we provided actionable insights for improving communications with potential participants, supporting primary care referral, honing the delivery model with better provider relationships and more patient choice, increasing understanding of behaviour change techniques, and enriching the educational and health coaching content.


Optimization of Solid-State Fermentation for Improved Conidia Production of Beauveria bassiana as a Mycoinsecticide.
The production of conidia of entomopathogenic Beauveria bassiana by solid-state fermentation was studied for the development of a biocontrol agent against aphid Myzus persicae. The optimal conditions for conidia production on polished white rice were 40% moisture content, 25℃ culture temperature, 2-day-old seeding culture grown in 3% corn meal, 2% rice bran, 2% corn steep powder medium, initial conidia concentration of 10(7) conidia/g in the wet rice, 10% inoculum size, and use of a polyethylene bag as a container. The polyethylene bag containing inoculated rice was hand-shaken every 12 hr during fermentation. Using optimal conditions, the maximum conidia production obtained was 4.05 g conidia/100 g dry rice after 14 days of cultivation, a rate 2.83 times higher than conidia yield of pre-optimization.


The genes encoding the DNA binding protein and the 23K protease of adenovirus types 40 and 41.
The adenovirus (Ad) single-stranded DNA binding protein (DBP) is a multifunctional protein. It is thought to consist of two domains, the amino-terminal domain involved in host-range determination and the carboxyl-terminal domain functioning in DNA replication and DNA binding. We have determined the nucleotide sequences of the DBP genes of Ad40 and Ad41, two human adenoviral serotypes that differ significantly from other adenoviruses. Regions of structural and functional importance in the corresponding proteins could be identified by comparison of the amino acid sequences with those of other known DBPs. In addition, the nucleotide sequences of the DBP early promoters, of the 23K protease genes, and of parts of the hexon and 100K protein genes have been determined. It can be deduced from the nucleotide sequences, that the Ad40 and Ad41 DBPs are relatively small (473 and 474 amino acids (a.a.), respectively, versus 529 a.a. for the Ad5 DBP). This is caused by the presence of very small amino-terminal domains of 119 a.a. (Ad40) and 120 a.a. (Ad41), as compared to 173 a.a. for the corresponding Ad5 domain. Only a few amino acids in this domain have been conserved in all known DBPs. The carboxyl-terminal domains show a higher degree of sequence conservation. In this domain, four strongly conserved regions can be identified, one of which might form a metal-binding site. The 23K proteases of both Ad40 and Ad41 show a strong homology to the Ad2 and Ad5 proteins, with the exception of the carboxyl-terminal end of the proteins. The 23K protease gene of Ad41 has an open reading frame that extends beyond the polyadenylation signal, in contrast to the Ad40 gene that ends well in front of the signal.


The psbC start codon in Synechocystis sp. PCC 6803.
The translation start codon for psbC, the gene encoding CP43, a chlorophyll-binding protein of photosystem II, has been identified for the cyanobacterium Synechosystis sp. PCC 6803 using site-directed mutagenesis. An AUG codon, about 50 bases upstream from the end of psbD-I had previously been assumed to be the translation start site of psbC. However, the fact that the AUG codon is not present in psbC from several other organisms, whereas a GUG codon 14 bases upstream from the end of psbD-I is strictly conserved suggests that CP43 translation starts at the latter codon. Mutation of GUG, but not of AUG, led to a loss of CP43 and photoautotrophic growth, indicating that the GUG codon is the sole initiation site for translation of the CP43 protein in Synechocystis sp. PCC 6803.


Garden-Hose Mitral Regurgitation: A Variant That Can Result in Underestimation of Severity: A Multimodality Imaging Case Study.
The quantitative assessment of mitral regurgitation (MR) by echocardiography has limitations. Cardiac magnetic resonance (CMR) imaging has an emerging role in the quantitation of MR, and preliminary studies indicate that CMR assessment may more accurately quantify MR and better correlate with postsurgical left ventricular reverse remodeling. The authors here report a case of MR in which multimodality imaging with CMR and transesophageal echocardiography was crucial in accurately diagnosing the severity of MR when transthoracic and provocative supine bike echocardiography underestimated the degree of MR in a unique variant known as "garden-hose" MR.


Digital mobile telephones and interference of ophthalmic equipment.
To assess the effect of mobile telephone electromagnetic interference on electronic ophthalmic equipment. Prospective audit with mobile telephones placed at distances of 3 m, 1 m, and 30 cm from, and in contact with, electronic ophthalmic equipment. Any interruption or cessation of the function of the ophthalmic device was assessed with the mobile telephones in standby, and in dialling or receiving modes. Any alterations of displayed digital figures or numbers were also assessed. A total of 23 electronic ophthalmic devices in two hospital ophthalmology outpatient departments were evaluated. All six mobile telephones used, and 22 (95.7%) of the 23 ophthalmic equipment evaluated had the Conformité Européene (CE) mark. No device showed any interruption or cessation of function. There were no alterations of displayed digital figures or numbers. The only effect of any kind was found with four instruments (1 non-CE marked), where there was temporary flickering on the screen, and only occurred when the mobile telephones were dialling or receiving at a distance of 30 cm or less from the instruments. This study shows that among the electronic ophthalmic devices tested, none suffered failure or interruption of function, from mobile telephone interference. Although not comprehensive for all ophthalmic equipment, the results question the need for a complete ban of mobile telephones in ophthalmic departments. It highlights the need for a controlled, objectively measured study of the clinically relevant effects of mobile telephones in the ophthalmology outpatient setting.


Expression pattern of the orphan receptor LGR4/GPR48 gene in the mouse.
Leucine-rich G-protein-coupled Receptors (LGR) constitute a subfamily of receptors related to glycoprotein hormone receptors. Amongst them, LGR4, LGR5 and LGR6 form a cluster for which natural agonists are still unknown. By an extensive gene trapping approach, Leighton et al. (2001) obtained a mouse line in which the LGR4 gene is disrupted by a trap vector carrying two biological markers, beta-geo (a fusion between bacterial beta-galactosidase and neomycin phosphotransferase) and a placental alkaline phosphatase (PLAP). Due to perinatal lethality, characterization of adult mice homozygous for this insertion has been impaired. In the present study we have investigated LacZ and PLAP activity patterns in heterozygous mice as a marker for LGR4 natural expression at both macroscopic and histological levels. We present a detailed atlas of LGR4 expression, which displays very wide expression with particularly strong activity in cartilages, kidneys, reproductive tracts and nervous system cells.


Morphological and molecular characterization of Bambusa tulda with a note on flowering.
Flowering incidence in Bambusa tulda has a high socio-economic impact. The aim of the present study was to describe the species for morphological characters (vegetative and reproductive) as well as molecular markers in order to enable species identification at various stages of the life cycle. Thirty-two key morphological characters (15 culm and 17 culm-sheath) were studied along with detailed inflorescence and floral characters. Incidence of sporadic flowering was recorded. Genomic DNA was isolated from leaves collected from 17 eco-geographical locations and RAPD profiles were generated. The description of culm, culm-sheath, inflorescence and floral morphology are in agreement with the prior taxonomic description by Gamble in 1896, but in this communication a more detailed description and illustrations are presented. No seed set was recorded following sporadic flowering, probably due to prezygoting isolating mechanisms (herkogamy or protandry). All 17 populations surveyed generated identical RAPD profiles. Sporadic flowering may occur in B. tulda, but may not necessarily be followed by gregarious flowering, and does not result in seed production.