Bovine enterovirus 2: complete genomic sequence and molecular modelling of a reference strain and a wild-type isolate from endemically infected US cattle.
Bovine enteroviruses are members of the family Picornaviridae, genus Enterovirus. Whilst little is known about their pathogenic potential, they are apparently endemic in some cattle and cattle environments. Only one of the two current serotypes has been sequenced completely. In this report, the entire genome sequences of bovine enterovirus 2 (BEV-2) strain PS87 and a recent isolate from an endemically infected herd in Maryland, USA (Wye3A) are presented. The recent isolate clearly segregated phylogenetically with sequences representing the BEV-2 serotype, as did other isolates from the endemic herd. The Wye3A isolate shared 82 % nucleotide sequence identity with the PS87 strain and 68 % identity with a BEV-1 strain (VG5-27). Comparison of BEV-2 and BEV-1 deduced protein sequences revealed 72-73 % identity and showed that most differences were single amino acid changes or single deletions, with the exception of the VP1 protein, where both BEV-2 sequences were 7 aa shorter than that of BEV-1. Homology modelling of the capsid proteins of BEV-2 against protein database entries for picornaviruses indicated six significant differences among bovine enteroviruses and other members of the family Picornaviridae. Five of these were on the 'rim' of the proposed enterovirus receptor-binding site or 'canyon' (VP1) and one was near the base of the canyon (VP3). Two of these regions varied enough to distinguish BEV-2 from BEV-1 strains. This is the first report and analysis of full-length sequences for BEV-2. Continued analysis of these wild-type strains should yield useful information for genotyping enteroviruses and modelling enterovirus capsid structure.


An In Vitro Model to Probe the Regulation of Adipocyte Differentiation under Hyperglycemia.
The aim of this study was an in vitro investigation of the effect of high glucose concentration on adipogenesis, as prolonged hyperglycemia alters adipocyte differentiation. 3T3-L1 preadipocytes differentiated in the presence of varying concentrations of glucose (25, 45, 65, 85, and 105 mM) were assessed for adipogenesis using AdipoRed (Lonza) assay. Cell viability and proliferation were measured using MTT reduction and [(3)H] thymidine incorporation assay. The extent of glucose uptake and glycogen synthesis were measured using radiolabelled 2-deoxy-D-[1-(3)H] glucose and [(14)C]-UDP-glucose. The gene level expression was evaluated using reverse transcription-polymerase chain reaction and protein expression was studied using Western blot analysis. Glucose at 105 mM concentration was observed to inhibit adipogenesis through inhibition of CCAAT-enhancer-binding proteins, sterol regulatory element-binding protein, peroxisome proliferator-activated receptor and adiponectin. High concentration of glucose induced stress by increasing levels of toll-like receptor 4, nuclear factor κB and tumor necrosis factor α thereby generating activated preadipocytes. These cells entered the state of hyperplasia through inhibition of p27 and proliferation was found to increase through activation of protein kinase B via phosphoinositide 3 kinase dependent pathway. This condition inhibited insulin signaling through decrease in insulin receptor β. Although the glucose transporter 4 (GLUT4) protein remained unaltered with the glycogen synthesis inhibited, the cells were found to exhibit an increase in glucose uptake via GLUT1. Adipogenesis in the presence of 105 mM glucose leads to an uncontrolled proliferation of activated preadipocytes providing an insight towards understanding obesity.


Disparities in bone density across contemporary Amazonian forager-horticulturalists: Cross-population comparison of the Tsimane and Shuar.
This study investigates bone density across the life course among Bolivian Tsimane and Ecuadorian Shuar of Amazonia. Both groups are rural, high-fertility forager-horticulturalists, with high lifetime physical activity levels. We test whether Tsimane and Shuar bone density patterns are different from each other, and if both groups are characterized by lower osteoporosis risk compared to U.S. references. Anthropometric and calcaneal bone density data, obtained via quantitative ultrasonometry (QUS), were collected from 678 Tsimane and 235 Shuar (13-92 years old). Population and sex differences in QUS values (estimated bone mineral density, speed of sound, broadband ultrasound attenuation) by age group were assessed using Mann-Whitney U tests. Age-related change and age at peak QUS value were determined using polynomial regressions. One-way analyses of covariance assessed population-level differences in QUS values by age group adjusting for body mass index. Participants aged 50+ years at elevated osteoporosis risk were identified using a T score < -1.8; binomial tests assessed risk compared to U.S. references. Shuar males and females <50 years old have QUS values 3-36% higher than Tsimane, with differences evident in adolescence. Among Tsimane and Shuar, 49 and 23% of participants aged 50+ years old, respectively, are at high risk for osteoporosis, compared to 34% of Americans; Shuar osteoporosis risk is comparable to Americans, while Tsimane risk is elevated. Disparate patterns in QUS values are documented for Tsimane and Shuar, with pronounced differences early in life. Potential explanations for differences include gene-environment interactions and/or degree of market integration, which influences diet, activity profiles, pathogen exposures, and other lifestyle covariates. As Tsimane osteoporosis risk is greater than in the United States, findings point to alternative risk factors for low bone density that are not readily discernible in industrialized populations.


[Amiodarone-associated thyroid dysfunction: prevalence and possibilities of correction].
During 1 year of amiodarone intake development of amiodarone-associated thyroid dysfunction was observed in 25% of patients (hypothyroidism and thyrotoxicosis in 19.2 and 5.8%, respectively). Development of hypothyroidism was not accompanied with loss of antiarrhythmic efficacy of amiodarone and therapy with L-thyroxin was conducted at the background of continued amiodarone intake. In all patients with clinical and in less than one half (47.6%) of patients with subclinical forms of hypothyroidism replacement therapy with L-thyroxin was carried out. Development of amiodarone-associated thyrotoxicosis was accompanied with loss of antiarrhythmic efficacy of amiodarone in all cases. In all patients with thyrotoxicosis which developed during amiodarone intake thyrostatic therapy with mercasolil was carried out and in case of its inefficacy prednisolone was added. In 87.5% of patients with thyrotoxicosis correction of the thyroid status was conducted under conditions of continued amiodarone intake as this drug had been prescribed because of life saving indications. Achievement of euthyroid state was followed by restoration of antiarrhythmic efficacy of amiodarone. Amiodarone was discontinued just in 1 patient with ventricular extrasystole as correction of thyroid status and restoration of euthyroidosis enabled effective use of other antiarrhythmic drugs.


Multiple measurement of serum lipids in the elderly.
Although there is considerable interest in underutilization of lipid testing, little is known about the prevalence and factors associated with overtesting of serum lipids. We assessed the number of different days in which outpatient lipid testing was performed in a 5% national sample of patients with parts A and B Medicare in 2006. Covariates included patient characteristics (age, race, prior diagnosis of lipid disorder, and other indications for lipid testing), number of usual care physicians (UCP), type of UCP, total outpatient physician encounters, and health referral region (HRR) characteristics (average per-patient Medicare expenditures and percent of patients seeing multiple UCPs). Among the 1,151,891 patients, 11.9% underwent 3 or more outpatient measurements of serum lipids. In multivariable analyses, the total number of UCPs providing care for the patient was associated with multiple lipid testing, independent of patient characteristics, indications for lipid testing, and total outpatient encounters. There was a strong association among HRRs between the rate of multiple lipid testing and average Medicare expenditures (r = 0.56). This was reduced after including the percentage of patients with more than 2 medical subspecialist UCPs in the HRR in a partial correlation (r = 0.31). Multiple lipid testing is associated with the presence of multiple providers, independent of indications for testing, comorbidity, and total physician visits. Much of the association of multiple lipid testing with medical expenditures at the level of HRR appears to be explained by differences in exposure to multiple providers.


Sensitized near-IR luminescence of lanthanide complexes based on push-pull diketone derivatives.
Lanthanide complexes with two push-pull diketone derivatives as sensitizers have been developed as synthons for near-infrared emitting materials. The ligand substituents consist of a carbazole moiety with hole-transport properties and an aromatic or heteroaromatic unit. According to quantitative NMR analysis and complementary HPLC experiments, the diketones are predominantly in their enolic form in polar solvents such as THF and MeCN at room temperature. The preferred cis-enol form contributes strongly to the binding of lanthanide ions (Ln = Nd, Gd, Er). The resulting tris(diketonate) ternary complexes with terpyridine (Ln = Nd, Er) display sizeable near-IR emission with long luminescence lifetimes.


Newborn Screening for Presymptomatic Diagnosis of Complement and Phagocyte Deficiencies.
The clinical outcomes of primary immunodeficiencies (PIDs) are greatly improved by accurate diagnosis early in life. However, it is not common to consider PIDs before the manifestation of severe clinical symptoms. Including PIDs in the nation-wide newborn screening programs will potentially improve survival and provide better disease management and preventive care in PID patients. This calls for the detection of disease biomarkers in blood and the use of dried blood spot samples, which is a part of routine newborn screening programs worldwide. Here, we developed a newborn screening method based on multiplex protein profiling for parallel diagnosis of 22 innate immunodeficiencies affecting the complement system and respiratory burst function in phagocytosis. The proposed method uses a small fraction of eluted blood from dried blood spots and is applicable for population-scale performance. The diagnosis method is validated through a retrospective screening of immunodeficient patient samples. This diagnostic approach can pave the way for an earlier, more comprehensive and accurate diagnosis of complement and phagocytic disorders, which ultimately lead to a healthy and active life for the PID patients.


Gene identification and structural characterization of the pyridoxal 5'-phosphate degradative protein 3-hydroxy-2-methylpyridine-4,5-dicarboxylate decarboxylase from mesorhizobium loti MAFF303099.
The function of the mlr6791 gene from Mesorhizobium loti MAFF303099 has been identified. This gene encodes 3-hydroxy-2-methylpyridine-4,5-dicarboxylate decarboxylase (HMPDdc), an enzyme involved in the catabolism of pyridoxal 5'-phosphate (Vitamin B6). This enzyme was overexpressed in Escherichia coli and characterized. HMPDdc is a 26 kDa protein that catalyzes the decarboxylation of 3-hydroxy-2-methylpyridine-4,5-dicarboxylate to 3-hydroxy-2-methylpyridine-5-carboxylate. The KM and kcat were found to be 366 microM and 0.6 s-1, respectively. The structure of this enzyme was determined at 1.9 A resolution using SAD phasing and belongs to the class II aldolase/adducin superfamily. While the decarboxylation of hydroxy-substituted benzene rings is a common motif in biosynthesis, the mechanism of this reaction is still poorly characterized. The structural studies described here suggest that catalysis of such decarboxylations proceeds by an aldolase-like mechanism.


Pathophysiology of ischaemic heart disease.
To summarize recent findings in the pathogenesis of ischemic heart disease (IHD) in people living with HIV (PLWH). PLWH have an elevated risk of IHD. Although incidence is declining, this condition still represents a major cause of non-AIDS-related mortality. The cause is likely multifactorial: traditional risk factors play an important role and IHD risk might be reduced with greater emphasis on primary prevention. The contribution of specific antiretroviral agents to IHD risk is changing as antiretroviral coverage increases globally and as safer agents have replaced drugs with well-described metabolic toxicities. The beneficial impact of virological suppression on antiretroviral therapy (ART) in reducing IHD is particularly evident in participants with advanced HIV infection and high baseline cardiovascular risk. The association between current abacavir use and myocardial infarction is still unexplained and indicates that mechanisms other than metabolic alterations may underlie IHD in PLWH. Consequently, the contributions of inflammation, subclinical atherosclerosis and endothelial dysfunction are receiving greater attention. Modern ART coupled with intensified efforts towards primary prevention is the cornerstone of IHD risk management in PLWH. The role of chronic inflammation and its optimal management need to be defined.


Prevalence and correlates of asthma in children with internalizing psychopathology.
Our objective was to determine the prevalence rate of parent-reported asthma in children with internalizing disorders seeking psychological treatment, and to study the level of internalizing and externalizing problems in these patients compared to patients without asthma. Participants were 367 children (ages 5-18 years) with internalizing disorders seeking psychological treatment. Children's psychiatric diagnosis was established with the Anxiety Disorders Interview Schedule for DSM-IV-Child and Parent versions. Parents reported on their child's asthma diagnosis, medical history, and medication usage. Child psychopathology was assessed with the Child Behavior Checklist and by child self-report with the Multidimensional Anxiety Scale for Children and the Children's Depression Inventory. We assessed internalizing psychopathology of the mothers with the Depression Anxiety and Stress Scale. An additional diagnosis of parent-reported asthma was established for 15% of the children diagnosed with an Axis I internalizing disorder, a prevalence rate markedly higher than reported for current parent-reported childhood asthma in the U.S. population. Patients with asthma showed higher levels of internalizing problems than their nonasthmatic counterparts. Internalizing psychopathology was not higher for mothers of patients with asthma. Asthma is a significant problem within the population of patients with childhood internalizing disorders. It can be accompanied by a greater severity of internalizing problems and may require specific precautions in the treatment protocol. Though parent report of asthma diagnosis is commonly used in surveys of childhood asthma, our findings have to be viewed in the light of its limitations.


Overview of the Office of Research, Statistics, and International Policy functions.
Research and statistics have been an integral part of social security program development and administration since the program's beginning. The Office of Research, Statistics, and International Policy (ORSIP) is responsible for developing and disseminating information on the effects of programs operated by the Social Security Administration (SSA) and the interactions among these programs, other tax and transfer programs, and economic, social, and demographic forces. This mission is carried out by collecting and tabulating statistics and by conducting policy research and legislative impact analyses. ORSIP also provides technical assistance to other SSA components. In this overview, each of those functions is outlined and major initiatives to carry out the office's primary objectives for fiscal year 1986 are described.


Hyperoside attenuates OVA-induced allergic airway inflammation by activating Nrf2.
Allergic airways disease (AAD) is one of the most common medical illnesses that is associated with an increased allergic airway inflammation. Hyperoside, an active compound isolated from Rhododendron brachycarpum G. Don, has been reported to have anti-inflammatory effect. The aim of this study was to analyze the protective effect of hyperoside on OVA-induced allergic airway inflammation in mice. In the present study, the mouse asthma model was induced by given OVA and hyperoside was administrated 1h before OVA challenge. The levels of IL-4, IL-5, IL-13, and IgE were detected by ELISA. H&E staining was used to assess lung histopathological changes. The expression of NF-κB p65, IκB, HO-1, and Nf-E2 related factor 2 (Nrf2) were measured by western blot analysis. The results showed that hyperoside significantly reduced the inflammatory cells infiltration and the levels of IL-4, IL-5, IL-13, and IgE. Hyperoside significantly inhibited OVA-induced oxidative stress as demonstrated by decreased MDA, and increased GSH and SOD levels. Treatment of hyperoside also inhibited OVA-induced airway hyperresponsiveness (AHR). Furthermore, the results showed that treatment of hyperoside significantly inhibited LPS-induced NF-κB activation. In addition, hyperoside was found to activate Nrf2/HO-1 signaling pathway. In conclusion, these results suggest that hyperoside ameliorates OVA-induced allergic airway inflammation by activating Nrf2 signaling pathway.


Monoclonal antibody to human esophageal cancer endothelium inhibits angiogenesis and tumor growth.
Monoclonal antibodies to tumor endothelial cells (TECs) hold great promise for cancer angiogenesis-targeted therapy. The aim of the present study was to develop such an agent for esophageal cancer treatment. BALB/c mice were immunized with human esophageal tumor endothelial cells (ETECs) cultured with tumor homogenate. MAbs were produced, screened by immunofluorescence and immunohistochemistry (IHC) and an IgG1K mAb 4B3 was selected. The mAb 4B3 antigen was analyzed by IHC and Western blotting. The antibody's effects on ETECs were determined by adhesion and tube formation assays, while its therapeutic potential was evaluated with a tumor model established by co-inoculating mice with the human esophageal cancer cell lines KYSE180 and ETECs. MAb 4B3 recognized a 40-kDa surface antigen preferentially expressed on TECs and other stromal cells in human malignant tissues of esophagus, stomach, colon, liver, lung and breast compared with their normal counterparts. The antigen was not detected on cancer cells or normal epithelia in these tissues, nor was it detectable on any cells in the mouse xenografts of KYSE180, including the host endothelia. MAb 4B3 inhibited ETEC adhesion to extracellular matrix proteins and tube formation in vitro. The antibody inhibited angiogenesis and growth of the tumor formed by coinoculation. These results suggest that mAb 4B3 has therapeutic potential for esophageal cancer.


Morphological and molecular changes in the Harderian gland of streptozotocin-induced diabetic rats.
Using a rat model of type 1 diabetes (T1D) obtained by treatment with streptozotocin, an antibiotic that destroys pancreatic β-cells, we evaluated the influence of subsequent hyperglycemia on the morphology and physiology of the Harderian gland (HG). HG is located in the medial corner of the orbit of many terrestrial vertebrates and, in rodents, is characterized by the presence of porphyrins, which being involved in the phototransduction, through photo-oxidation, produce reactive oxygen species activating the autophagy pathway. The study focused on the expression of some morphological markers involved in cell junction formation (occludin, connexin-43, and α-tubulin) and mast cell number (MCN), as well as autophagic and apoptotic pathways. The expression of enzymes involved in steroidogenesis [steroidogenic acute regulatory protein (StAR), and 3β-hydroxysteroid dehydrogenase (3β-HSD)] and the level of lipid peroxidation by thiobarbituric acid reactive species assay were also evaluated. The results strongly indicate, for the first time, that T1D has a negative impact on the pathophysiology of rat HG, as evidenced by increased oxidative stress, morphological and biochemical alterations, hyperproduction and secretion of porphyrins, increased MCN, reduced protein levels of StAR and 3β-HSD, and, finally, induced autophagy and apoptosis. All the combined data support the use of the rat HG as a suitable experimental model to elucidate the molecular damage/survival pathways elicited by stress conditions.


A role for serotonin in the hypothalamic-pituitary-adrenal response to insulin stress.
Controversy exists concerning the possible involvement of serotonin (5-HT) in the pituitary-adrenocortical response to stress. In the present research, a variety of pharmacological and physiological manipulations were used in male rats to study the role of this neurotransmitter in the adrenocortical response to insulin-induced hypoglycemia. We first examined the effect of insulin stress on hypothalamic 5-HT metabolism and found increased turnover as determined by an enhanced accumulation of 5-HT following monoamine oxidase inhibition. Brain 5-HT depletion by intraventricular injection of 5,7-dihydroxytryptamine significantly attenuated the corticosterone response to insulin, while treatment with the 5-HT receptor blocker methysergide tended to have the same effects. The corticosterone response to insulin was potentiated by prior administration of L-tryptophan, and blocked by pretreatment with valine, an amino acid that competes with tryptophan for transport across the blood-brain barrier. It therefore appears that the pituitary-adrenal response to insulin is mediated at least in part by 5-HT, and may be dependent on increased uptake of tryptophan by the brain.


Antimalarial efficacy of hydroxyethylapoquinine (SN-119) and its derivatives.
Quinine and other cinchona-derived alkaloids, although recently supplanted by the artemisinins (ARTs), continue to be important for treatment of severe malaria. Quinine and quinidine have narrow therapeutic indices, and a safer quinine analog is desirable, particularly with the continued threat of antimalarial drug resistance. Hydroxyethylapoquinine (HEAQ), used at 8 g a day for dosing in humans in the 1930s and halving mortality from bacterial pneumonias, was shown to cure bird malaria in the 1940s and was also reported as treatment for human malaria cases. Here we describe synthesis of HEAQ and its novel stereoisomer hydroxyethylapoquinidine (HEAQD) along with two intermediates, hydroxyethylquinine (HEQ) and hydroxyethylquinidine (HEQD), and demonstrate comparable but elevated antimalarial 50% inhibitory concentrations (IC50) of 100 to 200 nM against Plasmodium falciparum quinine-sensitive strain 3D7 (IC50, 56 nM). Only HEAQD demonstrated activity against quinine-tolerant P. falciparum strains Dd2 and INDO with IC50s of 300 to 700 nM. HEQD had activity only against Dd2 with an IC50 of 313 nM. In the lethal mouse malaria model Plasmodium berghei ANKA, only HEQD had activity at 20 mg/kg of body weight comparable to that of the parent quinine or quinidine drugs measured by parasite inhibition and 30-day survival. In addition, HEQ, HEQD, and HEAQ (IC50 ≥ 90 μM) have little to no human ether-à-go-go-related gene (hERG) channel inhibition expressed in CHO cells compared to HEAQD, quinine, and quinidine (hERG IC50s of 27, 42, and 4 μM, respectively). HEQD more closely resembled quinine in vitro and in vivo for Plasmodium inhibition and demonstrated little hERG channel inhibition, suggesting that further optimization and preclinical studies are warranted for this molecule.


Genetic variation in mutualistic and antagonistic interactions in an invasive legume.
Mutualists may play an important role in invasion success. The ability to take advantage of novel mutualists or survive and reproduce despite a lack of mutualists may facilitate invasion by those individuals with such traits. Here, we used two greenhouse studies to examine how soil microbial communities in general and mutualistic rhizobia in particular affect the performance of a legume species (Medicago polymorpha) that has invaded five continents. We performed two plant growth experiments with Medicago polymorpha, inoculating them with soil slurries in one experiment or rhizobial cultures in another experiment. For both experiments, we compared the growth of Medicago in competition with conspecific or heterospecific plants and examined variation among plant genotypes collected from the native and introduced ranges. We found that all genotypes experienced similar increases in biomass and formed more nodules that house rhizobia bacteria when inoculated with soil from a previously invaded site, compared to uninoculated plants or plants inoculated with soil from uninvaded and low invasion sites. In a second experiment, plants inoculated with rhizobia generally produced more biomass, had greater tolerance to interspecific competition, and had greater effects on competitor biomass than uninoculated plants. However, plant genotypes collected from the native range benefited more from rhizobia and were less tolerant of competition relative to genotypes collected from the introduced range. In the introduced range, compatible mutualists may not be readily available but competition is intense, causing Medicago to evolve to benefit less from interactions with rhizobia mutualists, while simultaneously becoming more tolerant of competition.


Sarcoidosis of the upper and lower airways.
Sarcoidosis is a systemic granulomatous disease of undetermined etiology characterized by a variable clinical presentation and disease course. Although clinical granulomatous inflammation may occur within any organ system, more than 90% of sarcoidosis patients have lung disease. Sarcoidosis is considered an interstitial lung disease that is frequently characterized by restrictive physiologic dysfunction on pulmonary function tests. However, sarcoidosis also involves the airways (large and small), causing obstructive airways disease. It is one of a few interstitial lung diseases that affects the entire length of the respiratory tract - from the nose to the terminal bronchioles - and causes a broad spectrum of airways dysfunction. This article examines airway dysfunction in sarcoidosis. The anatomical structure of the airways is the organizational framework for our discussion. We discuss sarcoidosis involving the nose, sinuses, nasal passages, larynx, trachea, bronchi and small airways. Common complications of airways disease, such as, atelectasis, fibrosis, bullous leions, bronchiectasis, cavitary lesions and mycetomas, are also reviewed.


Metal Contents of Lake Fish in Area Close to Disposal of Industrial Waste.
This research was conducted to analyze the content of Fe, Cu, Cd, Cr, and Pb in several species of fish taken from three lakes that are close to the disposal of industrial waste in Indonesia. The fish samples were taken from three lakes, namely, Muara Angke, Weda, and Morowali. The samples from Morowali were analyzed in April 2019, those from Weda from November to December 2019, and those from Muara Angke in June 2018. All the samples were then analyzed at the Chemistry Laboratory of the Department of Chemistry, Faculty of Mathematics and Natural Science, University of Indonesia, and the Integrated Laboratory of IPB. The main results showed that all types of fish from Morowali and Weda were no longer safe to consume because they contained Fe, Cu, Cd, and Cr exceeding the threshold of metal contamination. Meanwhile, all types of fish from Muara Angke, except for ayam-ayam, are still safe for consumption. The results of this study can be a source of information regarding metal content in fish and fish feed for safe consumption. Given the high consumption rate of fish and the hazards of heavy metals on humans' health, such research must be furthered.


Susceptibility of Clinical Isolates of Escherichia coli to Fosfomycin as Measured by Four In Vitro Testing Methods.
Clinical isolates of Escherichia coli (n = 554) were tested against fosfomycin using agar dilution, disk diffusion, and Etest. Agar dilution (reference method) identified few isolates with fosfomycin MICs of 64 (n = 3), 128 (n = 4), and ≥256 μg/ml (n = 2). Applying CLSI (M100, 2020) and EUCAST (v. 10.0, 2020) breakpoints, 98.9% and 98.4% (agar dilution), 99.3% and 99.1% (disk diffusion), and 99.1% and 98.9% (Etest) of isolates were fosfomycin susceptible, respectively. Essential agreement (agar dilution versus Etest) was low (40.8%); 59.3% (131/221) of isolates with agar dilution MICs of 2 to 128 μg/ml tested 2 to 4 doubling dilutions lower by Etest. Applying CLSI breakpoints, categorical agreement was >99% for both disk diffusion and Etest; no major errors (MEs) or very major errors (VMEs) were identified, and rates of minor errors (mEs) were <1%. EUCAST breakpoints yielded categorical agreements of >99% and no MEs for both disk diffusion and Etest; however, VMEs occurred at unacceptable rates of 44.4% (disk diffusion) and 33.3% (Etest). All isolates with agar dilution MICs of ≥32 μg/ml (n = 12) and a subset of isolates with MICs of ≤16 μg/ml (n = 49) were also tested using the Vitek 2 AST-N391 card and generated fosfomycin MICs 1 to ≥3 doubling dilutions lower than agar dilution for 11/12 isolates with agar dilution MICs of ≥32 μg/ml. We conclude that performing fosfomycin disk diffusion or Etest on urinary isolates of E. coli and interpreting results using CLSI breakpoints reliably identified fosfomycin-susceptible isolates regardless of differences in endpoint reading criteria. EUCAST breakpoints generated excessive rates of VMEs for our isolate collection of high fosfomycin susceptibility.


Prevalence of Pulmonary Embolism Among Patients With COPD Hospitalized With Acutely Worsening Respiratory Symptoms.
The prevalence of pulmonary embolism in patients with chronic obstructive pulmonary disease (COPD) and acutely worsening respiratory symptoms remains uncertain. To determine the prevalence of pulmonary embolism in patients with COPD admitted to the hospital for acutely worsening respiratory symptoms. Multicenter cross-sectional study with prospective follow-up conducted in 7 French hospitals. A predefined pulmonary embolism diagnostic algorithm based on Geneva score, D-dimer levels, and spiral computed tomographic pulmonary angiography plus leg compression ultrasound was applied within 48 hours of admission; all patients had 3-month follow-up. Patients were recruited from January 2014 to May 2017 and the final date of follow-up was August 22, 2017. Acutely worsening respiratory symptoms in patients with COPD. The primary outcome was pulmonary embolism diagnosed within 48 hours of admission. Key secondary outcome was pulmonary embolism during a 3-month follow-up among patients deemed not to have venous thromboembolism at admission and who did not receive anticoagulant treatment. Other outcomes were venous thromboembolism (pulmonary embolism and/or deep vein thrombosis) at admission and during follow-up, and 3-month mortality, whether venous thromboembolism was clinically suspected or not. Among 740 included patients (mean age, 68.2 years [SD, 10.9 years]; 274 women [37.0%]), pulmonary embolism was confirmed within 48 hours of admission in 44 patients (5.9%; 95% CI, 4.5%-7.9%). Among the 670 patients deemed not to have venous thromboembolism at admission and who did not receive anticoagulation, pulmonary embolism occurred in 5 patients (0.7%; 95% CI, 0.3%-1.7%) during follow-up, including 3 deaths related to pulmonary embolism. The overall 3-month mortality rate was 6.8% (50 of 740; 95% CI, 5.2%-8.8%). The proportion of patients who died during follow-up was higher among those with venous thromboembolism at admission than the proportion of those without it at admission (14 [25.9%] of 54 patients vs 36 [5.2%] of 686; risk difference, 20.7%, 95% CI, 10.7%-33.8%; P < .001). The prevalence of venous thromboembolism was 11.7% (95% CI, 8.6%-15.9%) among patients in whom pulmonary embolism was suspected (n = 299) and was 4.3% (95% CI, 2.8%-6.6%) among those in whom pulmonary embolism was not suspected (n = 441). Among patients with chronic obstructive pulmonary disease admitted to the hospital with an acute worsening of respiratory symptoms, pulmonary embolism was detected in 5.9% of patients using a predefined diagnostic algorithm. Further research is needed to understand the possible role of systematic screening for pulmonary embolism in this patient population.


Effect of pepA deletion and overexpression in Aspergillus luchuensis on sweet potato shochu brewing.
The mash of sweet potato shochu (Japanese distilled spirit) has a low pH value because the shochu koji mold produces a large amount of citric acid, which prevents germ contamination. In this study, we examined acid protease PepA's role in shochu production. For this purpose, we constructed pepA deletion and overexpression strains, using a black koji mold Aspergillus luchuensis RIB 2604 (NBRC 4314), with the Agrobacterium-mediated transformation method. The rice koji, prepared using a pepA disruptant (ΔpepA) and pepA-overexpressing strain (OEpepA), demonstrated 1/2- and 24-fold acid protease activities compared to that prepared using the parental strain, respectively. A small-scale test of sweet potato shochu brewing indicated the mash of ΔpepA had a lower amino acid concentration, while the mash of OEpepA had a higher concentration than that produced by the parental strain. Therefore, the mash amino acid concentrations were proportional to these strains' acid proteases activities. After distilling these mashes, we examined each shochu's aroma components. Shochu prepared using ΔpepA had relatively higher aroma components, such as alcohol and ester, compared to that prepared using parental strains. Meanwhile, shochu prepared using OEpepA had lower aroma components than that prepared using the parental strains. Based on these results, the amount of shochu aroma components showed an inverse correlation to the acid protease activity in the mash. Thus, the koji mold's acid protease content had a greater influence on the aroma qualities of sweet potato shochu. Accordingly, we have discussed the possibility of the breeding of shochu koji mold with acid protease as an indicator.


Novel type of proliferating lymphoplasmacytoid cell with a characteristic spotted immunofluorescence pattern.
We examined bone marrow from myeloma patients for the presence of cells with the characteristics of the clonogenic cell in the myeloma stem cell assay. We identified a novel type of cell that contained cytoplasmic immunoglobulin of the relevant idiotype located in a cytoplasmic spot. This "spotted" Ig could be located in the rough endoplasmic reticulum. Spotted cells are highly proliferative, as evidenced by the nuclear staining with the antibody Ki67, and were found in the bone marrow from most of the myeloma patients studied. This type of cell was also present in patients with immunocytomas, in some cases of benign monoclonal gammopathy, and in patients in the state of polyclonal hypergammaglobulinemia. IgG subclass distribution of so-called spotted cells and plasma cells, found in a patient with pseudo biclonal gammopathy, indicates that spotted cells are intermediate between B cells and plasma cells. Spotted cells express the B cell-associated antigens HB4 and HB6 but do not express other B cluster of differentiation antigens or plasmacytoid antigens tested.


Defense mechanisms against herbivory in Picea: sequence evolution and expression regulation of gene family members in the phenylpropanoid pathway.
In trees, a substantial amount of carbon is directed towards production of phenolics for development and defense. This metabolic pathway is also a major factor in resistance to insect pathogens in spruce. In such gene families, environmental stimuli may have an important effect on the evolutionary fate of duplicated genes, and different expression patterns may indicate functional diversification. Gene families in spruce (Picea) have expanded to superfamilies, including O-methyltransferases, cytochrome-P450, and dirigents/classIII-peroxidases. Neo-functionalization of superfamily members from different clades is reflected in expression diversification. Genetical genomics can provide new insights into the genetic basis and evolution of insect resistance in plants. Adopting this approach, we merged genotype data (252 SNPs in a segregating pedigree), gene expression levels (for 428 phenylpropanoid-related genes) and measures of susceptibility to Pissodes stobi, using a partial-diallel crossing-design with white spruce (Picea glauca). Thirty-eight expressed phenylpropanoid-related genes co-segregated with weevil susceptibility, indicating either causative or reactive effects of these genes to weevil resistance. We identified eight regulatory genomic regions with extensive overlap of quantitative trait loci from susceptibility and growth phenotypes (pQTLs) and expression QTL (eQTL) hotspots. In particular, SNPs within two different CCoAOMT loci regulate phenotypic variation from a common set of 24 genes and three resistance traits. Pest resistance was associated with individual candidate genes as well as with trans-regulatory hotspots along the spruce genome. Our results showed that specific genes within the phenylpropanoid pathway have been duplicated and diversified in the conifer in a process fundamentally different from short-lived angiosperm species. These findings add to the information about the role of the phenylpropanoid pathway in the evolution of plant defense mechanisms against insect pests and provide substantial potential for the functional characterization of several not yet resolved alternative pathways in plant defenses.


Prenatal exposure to polycyclic aromatic hydrocarbons (PAHs) augments neonatal hyperoxic lung injury: Role of cytochrome P450 (CYP)1A1, 1A2, and 1B1.
Pregnant women exposed to polycyclic aromatic hydrocarbons (PAHs) are at increased risk for premature delivery. Premature infants often require supplemental oxygen, a known risk factor for bronchopulmonary dysplasia (BPD). Cytochrome P450 (CYP) enzymes have been implicated in hyperoxic lung injury. We hypothesize that prenatal PAH exposure exacerbates oxygen-mediated lung injury in neonatal mice, and that this effect is differentially altered in mice lacking the gene for (Cyp)1a1, 1a2, or 1b1. Timed pregnant wild type (WT) (C57BL/6J) mice were orally administered a PAH mixture of benzo[a]pyrene (BP) and benzo[b]fluoranthene (BbF) or the vehicle corn oil (CO) once daily on gestational days 16-19, and the dose response on postnatal lung injury was examined. In addition, timed pregnant mice with one of four genotypes, WT, Cyp1a1-null, Cyp1a2-null, and Cyp1b1-null, were treated orally with CO or PAH on gestational days 16-19 and exposed to hyperoxia or room air for 14 days. Lung injury was assessed on PND15 by radial alveolar count and mean linear intercept. Gene expression of DNA repair genes in lung and liver were measured. Results showed that hyperoxic lung injury is augmented by prenatal PAH exposure in a dose-dependent manner. This effect was differentially altered in the Cyp-null mice, with Cyp1a2-null showing the greatest extent of lung injury. We concluded that newborn mice exposed to PAH in utero had more significant lung injury in response to hyperoxia than non-PAH exposed pups, and CYP1A1 and CYP1A2 are protective against lung injury while CYP1B1 augments lung injury.


Nonfatal, unintentional, non--fire-related carbon monoxide exposures--United States, 2004-2006.
Carbon monoxide (CO) is a colorless, odorless, nonirritating gas that is produced through the incomplete combustion of hydrocarbons. Sources of CO include combustion devices (e.g., boilers and furnaces), motor-vehicle exhaust, generators and other gasoline or diesel-powered engines, gas space heaters, woodstoves, gas stoves, fireplaces, tobacco smoke, and various occupational sources. CO poisoning is a leading cause of unintentional poisoning deaths in the United States; it was responsible for approximately 450 deaths each year during 1999-2004 and an estimated 15,200 emergency department (ED) visits each year during 2001-2003. Health effects of CO exposure can range from viral-like symptoms (e.g., fatigue, dizziness, headache, confusion, and nausea) to more severe conditions (e.g., disorientation, unconsciousness, long-term neurologic disabilities, coma, cardiorespiratory failure, and death). CO poisoning often is misdiagnosed and underdetected because of the nonspecific nature of symptoms. To update a previously published report and provide national estimates of CO-related ED visits during 2004-2006, CDC analyzed data from the National Electronic Injury Surveillance System--All Injury Program (NEISS-AIP) database. During 2004-2006, an estimated average of 20,636 ED visits for nonfatal, unintentional, non-fire-related CO exposures occurred each year. Approximately 73% of these exposures occurred in homes, and 41% occurred during winter months (December-February). Prevention efforts targeting residential and seasonal CO exposures can substantially reduce CO-related morbidity.


Brief Isoflurane Anesthesia Produces Prominent Phosphoproteomic Changes in the Adult Mouse Hippocampus.
Anesthetics are widely used in medical practice and experimental research, yet the neurobiological basis governing their effects remains obscure. We have here used quantitative phosphoproteomics to investigate the protein phosphorylation changes produced by a 30 min isoflurane anesthesia in the adult mouse hippocampus. Altogether 318 phosphorylation alterations in total of 237 proteins between sham and isoflurane anesthesia were identified. Many of the hit proteins represent primary pharmacological targets of anesthetics. However, findings also enlighten the role of several other proteins-implicated in various biological processes including neuronal excitability, brain energy homeostasis, synaptic plasticity and transmission, and microtubule function-as putative (secondary) targets of anesthetics. In particular, isoflurane increases glycogen synthase kinase-3β (GSK3β) phosphorylation at the inhibitory Ser(9) residue and regulates the phosphorylation of multiple proteins downstream and upstream of this promiscuous kinase that regulate diverse biological functions. Along with confirmatory Western blot data for GSK3β and p44/42-MAPK (mitogen-activated protein kinase; reduced phosphorylation of the activation loop), we observed increased phosphorylation of microtubule-associated protein 2 (MAP2) on residues (Thr(1620,1623)) that have been shown to render its dissociation from microtubules and alterations in microtubule stability. We further demonstrate that diverse anesthetics (sevoflurane, urethane, ketamine) produce essentially similar phosphorylation changes on GSK3β, p44/p42-MAPK, and MAP2 as observed with isoflurane. Altogether our study demonstrates the potential of quantitative phosphoproteomics to study the mechanisms of anesthetics (and other drugs) in the mammalian brain and reveals how already a relatively brief anesthesia produces pronounced phosphorylation changes in multiple proteins in the central nervous system.


A Tunable Diffusion-Consumption Mechanism of Cytokine Propagation Enables Plasticity in Cell-to-Cell Communication in the Immune System.
Immune cells communicate by exchanging cytokines to achieve a context-appropriate response, but the distances over which such communication happens are not known. Here, we used theoretical considerations and experimental models of immune responses in vitro and in vivo to quantify the spatial extent of cytokine communications in dense tissues. We established that competition between cytokine diffusion and consumption generated spatial niches of high cytokine concentrations with sharp boundaries. The size of these self-assembled niches scaled with the density of cytokine-consuming cells, a parameter that gets tuned during immune responses. In vivo, we measured interactions on length scales of 80-120 μm, which resulted in a high degree of cell-to-cell variance in cytokine exposure. Such heterogeneous distributions of cytokines were a source of non-genetic cell-to-cell variability that is often overlooked in single-cell studies. Our findings thus provide a basis for understanding variability in the patterning of immune responses by diffusible factors.


The effect of trypsin digestion on the activities of polynucleotide phosphorylase.
1. Treatment of Micrococcus lysodeikticus polynucleotide phosphorylase (nucleoside diphosphate-polynucleotide nucleotidyltransferase) with trypsin causes a preferential loss of its cytidine diphosphate and uridine diphosphate polymerization activities. 2. The phosphorolytic activity of the enzyme towards polycytidylic acid is unaffected in conditions in which the cytidine diphosphate-polymerization activity without added primer is virtually abolished. 3. The treated enzyme retains its altered pattern of activities when purified fivefold by gel filtration. 4. The effect on the cytidine diphosphate-polymerization activity is due, in part, to a large increase in primer requirement as a result of proteolysis, and is qualitatively independent of the state of purity of the polynucleotide phosphorylase. 5. The enzyme is protected from trypsin degradation by nucleic acids, polynucleotides and nucleoside disphosphates. 6. A similar, but less marked differential effect, is caused by alpha-chymotrypsin.


Hierarchical coding of binary images.
Quadtrees are a compact hierarchical method of representation of images. In this paper, we explore a number of hierarchical image representations as applied to binary images, of which quadtrees are a single exemplar. We discuss quadtrees, binary trees, and an adaptive hierarchical method. Extending these methods into the third dimension of time results in several other methods. All of these methods are discussed in terms of time complexity, worst case and average compression of random images, and compression results on binary images derived from natural scenes. The results indicate that quadtrees are the most effective for two-dimensional images, but the adaptive algorithms are more effective for dynamic image sequences.


Immunocytochemical localization of interphotoreceptor retinoid-binding protein in developing normal and RCS rat retinas.
Interphotoreceptor retinoid-binding protein (IRBP) was localized immunocytochemically in developing normal and RCS rat retinas. IRBP was present in normal and RCS neural retinas on the day after birth (postnatal day 2, P2) to P8 in the space between the neuroblastic layer and the retinal pigment epithelium (RPE). The presence of IRBP prior to the development of outer segments (OS) suggests that OS formation is not linked temporally with IRBP secretion. On P10, staining was confined to the interphotoreceptor space with an intense band of label adjacent to the RPE. This staining pattern persisted in normal rats throughout development and until P18 in RCS rats. On P18, anti-IRBP staining in the RCS was spread evenly throughout the OS layer with no intense band of label adjacent to the RPE and after P18, there was decreased staining with anti-IRBP. On P45 and later, no staining of the RCS retina was found with anti-IRBP. Immunoblots of normal and RCS retinas corroborated the results from immunocytochemical staining. These findings suggest that IRBP may be synthesized in the photoreceptors, but is not abnormal in amount or distribution prior to onset of retinal degeneration in the RCS rat.


[The impact of researchers loyal to Big Pharma on the ethics and quality of clinical trials in Latin America].
This article explains the difficulties innovative pharmaceutical firms have in repaying shareholders with attractive dividends. The problem is the result of the expiration of the patents of blockbuster drugs and the difficulties that the firms have in bringing new blockbuster drugs to the market. One of the solutions companies have found has been to accelerate the implementation of clinical trials in order to expedite the commercialization of new drugs. Doing so increases the period in which they can sell drugs at monopoly prices. We therefore discuss how innovative pharmaceutical firms shorten the implementation time of clinical trials in Latin America and the consequences such actions have on the quality of the collected data, the protection of human rights of the subjects of experimentation, and compliance with the ethical principles approved in international declarations.


Exploiting the microbiome associated with normal and abnormal sprouting rice (Oryza sativa L.) seed phenotypes through a metabarcoding approach.
Rice germination and seedlings' growth are crucial stages that influence crop establishment and productivity. These performances depend on several factors, including the abundance and diversity of seed microbial endophytes. Two popular rainfed rice varieties cultivated in Cameroon, NERICA 3 and NERICA 8, were used for investigating the seed-associated microbiome using the Illumina-based 16 S rRNA gene. Significant differences were observed in terms of richness index between normal and abnormal seedlings developed from sprouting seeds, although no significant species evenness index was assessed within either phenotype. Two hundred ninety-two bacterial amplicon sequence variants were identified in seed microbiome of the rice varieties, and principal coordinate analysis revealed that microbial communities formed two distinct clusters in normal and abnormal seedling phenotypes. Overall, 38 bacteria genera were identified, belonging to 6 main phyla. Furthermore, the core microbiome was defined, and the differential abundance of 28 bacteria genera was assessed. Based on the collected results, putative bacterial genera were directly correlated with the development of normal seedlings. For most genera that are recognised to include beneficial species, such as Brevundimonas, Sphingomonas, Exiguobacterium, Luteibacter, Microbacterium and Streptomyces, a significant increase of their relative abundance was found in normal seedlings. Additionally, in abnormal seedlings, we also observed an increased abundance of the genera Kosakonia and Paenibacillus, which might have controversial aspects (beneficial or pathogenic), together with the presence of some genera (Clostridium sensu stricto) that are commonly correlated to sick plants. The putative functional gene annotation revealed the higher abundance of genes related to the metabolic biosynthesis of soluble carbohydrates and starch, tryptophan, nucleotides and ABC transporters in normal seedlings. Data presented in this study may help in further understanding the importance of the seed endophyte microbiome for driving a correct development of rice plants at the early stages and to identify possible beneficial bacteria for technological applications aimed to increase seed quality and crop productivity.


Bacteria of porcine skin, xenografts, and treatment with neomycin sulfate.
Homogenized 4-mm punch biopsies were taken from pigs and bacteriologically evaluated to determine the efficacy of surgical scrub procedures and the subsequent treatment of tissue with 0.5% neomycin sulfate-sodium bisulfite (neomycin-bisulfite) as a decontaminating agent. The majority of the lots of porcine skin taken directly from animals for xenografts in the treatment of burns contained viable bacteria at the time of grafting although scrubbing procedures substantially reduced the skin bacteria. The porcine bacteria consisted primarily of coagulase-negative staphylococci with most strains exhibiting caseinolytic and elastase activity. Staphylococci were the only abundant bacteria found in postscrub biopsies and in saline solutions used to wash the dermatome during its use. After an overnight exposure of grafting tissue soaked in neomycin-bisulfite, the spent neomycin-bisulfite solutions were tested for bacteriostatic and bactericidal activity by comparison to unused neomycin. All solutions tested were equal in bacteriostatic strength, but the bactericidal action of some spent solutions was decreased. Neomycin alone exerted a more lethal effect on sensitive bacteria than the neomycin-bisulfite solution. The desirability of having viable porcine skin for a xenograft necessitated using or discarding the tissue after storage in neomycin-bisulfite at 4 C for a maximum of 72 hr. Certain contaminating microorganisms were unaffected by antibiotic treatment, and the prolonged use of neomycin without bisulfite would have primarily eradicated only the porcine coagulase-negative staphylococci. Neither the presence of this group in grafting tissue nor their proteolytic activity had any observed adverse effect on xenografting success.


Detecting and Quantifying Natural Selection at Two Linked Loci from Time Series Data of Allele Frequencies with Forward-in-Time Simulations.
Recent advances in DNA sequencing techniques have made it possible to monitor genomes in great detail over time. This improvement provides an opportunity for us to study natural selection based on time serial samples of genomes while accounting for genetic recombination effect and local linkage information. Such time series genomic data allow for more accurate estimation of population genetic parameters and hypothesis testing on the recent action of natural selection. In this work, we develop a novel Bayesian statistical framework for inferring natural selection at a pair of linked loci by capitalising on the temporal aspect of DNA data with the additional flexibility of modeling the sampled chromosomes that contain unknown alleles. Our approach is built on a hidden Markov model where the underlying process is a two-locus Wright-Fisher diffusion with selection, which enables us to explicitly model genetic recombination and local linkage. The posterior probability distribution for selection coefficients is computed by applying the particle marginal Metropolis-Hastings algorithm, which allows us to efficiently calculate the likelihood. We evaluate the performance of our Bayesian inference procedure through extensive simulations, showing that our approach can deliver accurate estimates of selection coefficients, and the addition of genetic recombination and local linkage brings about significant improvement in the inference of natural selection. We also illustrate the utility of our method on real data with an application to ancient DNA data associated with white spotting patterns in horses.


Health burdens of surface ozone in the UK for a range of future scenarios.
Exposure to surface ozone (O3), which is influenced by emissions of precursor chemical species, meteorology and population distribution, is associated with excess mortality and respiratory morbidity. In this study, the EMEP-WRF atmospheric chemistry transport model was used to simulate surface O3 concentrations at 5km horizontal resolution over the British Isles for a baseline year of 2003, for three anthropogenic emissions scenarios for 2030, and for a +5°C increase in air temperature on the 2003 baseline. Deaths brought forward and hospitalisation burdens for 12 UK regions were calculated from population-weighted daily maximum 8-hour O3. The magnitude of changes in annual mean surface O3 over the UK for +5°C temperature (+1.0 to +1.5ppbv, depending on region) was comparable to those due to inter-annual meteorological variability (-1.5 to +1.5ppbv) but considerably less than changes due to precursor emissions changes by 2030 (-3.0 to +3.5ppbv, depending on scenario and region). Including population changes in 2030, both the 'current legislation' and 'maximum feasible reduction' scenarios yield greater O3-attributable health burdens than the 'high' emission scenario: +28%, +22%, and +16%, respectively, above 2003 baseline deaths brought forward (11,500) and respiratory hospital admissions (30,700), using O3 exposure over the full year and no threshold for health effects. The health burdens are greatest under the 'current legislation' scenario because O3 concentrations increase as a result of both increases in background O3 concentration and decreases in UK NOx emissions. For the +5°C scenario, and no threshold (and not including population increases), total UK health burden increases by 500 premature deaths (4%) relative to the 2003 baseline. If a 35ppbv threshold for O3 effects is assumed, health burdens are more sensitive to the current legislation and +5°C scenarios, although total health burdens are roughly an order of magnitude lower. In all scenarios, the assumption of a threshold increases the proportion of health burden in the south and east of the UK compared with the no threshold assumption. The study highlights that the total, and geographically-apportioned, O3-attributable health burdens in the UK are highly sensitive to the future trends of hemispheric, regional and local emissions of O3 precursors, and to the assumption of a threshold for O3 effect.


Pterygium-induced corneal astigmatism.
Previous work has suggested an association between increasing size of pterygium and increasing degrees of induced corneal astigmatism. To assess the quantitative relation between pterygium size and induced corneal astigmatism using a computerized corneal analysis system (TMS II) and slit-lamp beam evaluation of pterygium size, and to conclude whether corneal astigmatism is an early indication for surgical intervention. We evaluated 94 eyes of 94 patients with unilateral primary pterygium of different sizes, using TMS II and slit-lamp beam measurements of the size of the pterygium (in millimeters) from the limbus to assess parameters of pterygium size with induced corneal astigmatism. Best corrected visual Snellen acuity was performed. Primary pterygium induced with-the-rule astigmatism. Pterygium extending > 16% of the corneal radius or 1.1 mm or less from the limbus produced increasing degrees of induced astigmatism of more than 1.0 diopter. Significant astigmatism was found in 16.16% of 24 eyes with pterygium of 0.2 up to 1.0 mm in size, in 45.45% of 22 eyes with pterygium of 1.1 up to 3.0 mm in size (P < or = 0.0004), and in 100% of 3 eyes with pterygium of 5.1 up to 6.7 mm in size (P = 0.0005). We found that visual acuity was decreased when topographic astigmatism was increased. When primary pterygium reaches more than 1.0 mm in size from the limbus it induces with-the-rule significant astigmatism (> or = 1.0 diopter). This significant astigmatism tends to increase with the increasing size of the lesion. Topographic astigmatism tends to be improved by successful removal of the pterygium. These findings suggest that early surgical intervention in the pterygium may be indicated when the lesion is more than 1.0 mm in size from the limbus.


Long-term mortality following interhospital transfer for acute myocardial infarction.
Interhospital transfer of patients admitted with an acute myocardial infarction for specialised care is common and costly. However, the long-term mortality of transferred patients compared with patients solely treated at the presenting hospital has not been evaluated. Here, we assess the long-term mortality of patients who undergo interhospital transfer during their acute myocardial infarction admission. We evaluated 40 482 patients with a ICD10-AM diagnosis of acute myocardial infarction admitted to hospitals in New South Wales, Australia, from 2004 to 2008, of whom 10 107 (25%) were transferred. We compared in-hospital and mortality up to 5.5 years postdischarge among transferred and non-transferred patients. We created a 1:1 propensity score matched cohort (n=16 854; 8427 per group) to account for selection bias. In the matched cohort, transferred patients were more likely to undergo revascularisation (55.6% vs 13.7%, RR 4.05; 95% CI 3.83 to 4.29) and had lower mortality at 30 days (3.5% vs 5.7%, HR 0.60; 95% CI 0.52 to 0.70), 1 year (7.5% vs 12.6%, HR 0.58; 95% CI 0.52 to 0.64) and at the end of follow-up (15.3% vs 22.5%, HR 0.65; 95% CI 0.61 to 0.70) than patients treated in presenting hospitals. With the exception of transfers originating from revascularisation capable hospitals, these findings were consistent across a range of subgroups, including patients of all ages, ST-elevation myocardial infarction and non ST-elevation myocardial infarction patients, and transfers originating from hospitals in regional and major city areas. Sensitivity analyses showed that these findings are unlikely to be due to survival bias or to confounding by unmeasured variables. Patients hospitalised for an acute myocardial infarction who are transferred to one or more hospitals for specialised care have higher rates of coronary revascularisation and experience lower long-term mortality.


Swe1 regulation and transcriptional control restrict the activity of mitotic cyclins toward replication proteins in Saccharomyces cerevisiae.
Cyclin-dependent kinases (CDKs) drive the cell cycle through the phosphorylation of substrates that function in genome duplication and cell division. The existence of multiple cyclin subunits and their distinct cell cycle-regulated expression suggests that cyclins impart unique specificities to CDK-substrate interactions that are critical for normal cellular function. This study shows that the combination of early cell cycle expression and deletion of the CDK inhibitor Saccharomyces Wee1 (Swe1) enables the mitotic B-type (Clb) cyclins Clb2, Clb3, and Clb4 of Saccharomyces cerevisiae to initiate S phase with similar effectiveness as the S-phase cyclin Clb5. Although in vivo analysis indicates preferential phosphorylation of a replication substrate by Clb5-Cdk1, this difference is relatively minor compared with the impact of transcriptional control and Swe1 regulation. Indeed, early expressed Clb2-Cdk1 can activate all essential Clb-Cdk substrates in a strain lacking all other Clbs and Swe1. Thus, Swe1 regulation and expression timing are key mechanisms that sequester the broad activity of Clb2-Cdk1 from critical substrates. Furthermore, the ability of Swe1 to inhibit the activity of different B-type cyclins in replication initiation correlates with the normal expression timing of those cyclins, with no apparent in vivo inhibition of Clb5 and Clb6, moderate inhibition of Clb3 and Clb4, and strong inhibition of Clb2. Hence, Swe1 appears to reinforce the temporal activity of cyclins established through transcriptional control. The conserved nature of CDK function suggests that similar mechanisms regulate CDK specificity in multicellular organisms.


Spiritual coping predicts CD4-cell preservation and undetectable viral load over four years.
In this study of 177 people living with HIV, we examined if spiritual coping leads to slower HIV disease progression (CD4 cells, viral load [VL]), and more positive health behaviors (adherence, safer sex, less substance use). Prior research suggests that physicians' assessment of spiritual coping can be an interventional aid in promoting positive spiritual coping. Longitudinal spiritual coping was rated using qualitative content analysis of six-monthly interviews/essays. Positive spiritual coping (65%) was predominant over negative (7%), whereas 28% did not make significant use of spirituality as a means to cope. Spiritual coping was associated with less substance use disorder but not with less sexual risk behavior. Hierarchical linear modeling demonstrated that spiritual coping predicted sustained undetectable VL and CD4-cell preservation over four years, independent of sociodemographics, baseline disease status, and substance use disorder. Achieving undetectable VL significantly increased over time in participants with positive spiritual coping but decreased among those with negative spiritual coping. For every participant with positive spiritual coping achieving undetectable VL, four with negative spiritual coping reported with detectable/transmittable HIV. Notably, even when controlling for the effect of VL suppression, CD4-cell decline was 2.25 times faster among those engaged in negative versus positive spiritual coping. In conclusion, spiritual coping is associated with positive health behaviors, such as maintaining long-term VL suppression and less onset/relapse of substance use disorder over time. Among those who are sexually active, positive spiritual coping reduces the risk of HIV transmission via VL suppression but may not prevent the transmission of other STDs because spiritual coping is not related to safer sexual behavior. Notably, the association between spiritual coping and immune preservation was direct (i.e., not explained by VL suppression), suggesting potential psychoneuroimmunological pathways. Thus, assessment of spiritual coping may be an important area of intervention to achieve undetectable VL, reduce HIV disease progression, and prevent substance use onset/relapse.


Involvement of the plasma membrane Ca2+-ATPase in the short-term response of Arabidopsis thaliana cultured cells to oligogalacturonides.
Treatment of Arabidopsis thaliana cells with oligogalacturonides (OG) initiates a transient production of reactive oxygen species (ROS), the concentration of which in the medium peaks after about 20 min of treatment. The analysis of OG effects on Ca (2+) fluxes shows that OG influence both Ca (2+) influx and Ca (2+) efflux (measured as (45)Ca (2+) fluxes) in a complex way. During the first 10 - 15 min, OG stimulate Ca (2+) influx and decrease its efflux, while at successive times of treatment, OG cause an increase of Ca (2+) efflux and a slight decrease of its influx. Treatment with sub- micro M concentrations of eosin yellow (EY), which selectively inhibits the Ca (2+)-ATPase of plasma membrane (PM), completely prevents the OG-induced increase in Ca (2+) efflux. EY also suppresses the transient feature of OG-induced ROS accumulation, keeping the level of ROS in the medium high. The biochemical analysis of PM purified from OG-treated cells indicates that treatment with OG for 15 to 45 min induces a significant decrease in Ca (2+)-ATPase activation by exogenous calmodulin (CaM), and markedly increases the amount of CaM associated with the PM. During the same time span, OG do not influence the expression of At-ACA8, the main isoform of PM Ca (2+)-ATPase in suspension-cultured A. thaliana cells, and of CaM genes. Overall, the reported results demonstrate that the PM Ca (2+)-ATPase is involved in the response of plant cells to OG and is essential in regulation of the oxidative burst.


No Microscope? No Problem: A Systematic Review of Microscope-Free Microsurgery Training Models.
Benchtop microsurgical training models that use digital tools (smartphones, tablets, and virtual reality [VR]) for magnification are allowing trainees to practice without operating microscopes. This systematic review identifies existing microscope-free training models, compares models in their ability to enhance microsurgical skills, and presents a step-by-step protocol for surgeons seeking to assemble their own microsurgery training model. We queried PubMed, Embase, and Web of Science databases through November 2020 for microsurgery training models and performed a systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We collected data including training model characteristics (cost, magnification, and components) and outcomes (trainee satisfaction, image resolution, and faster suturing speed). We also conducted a complimentary Google search to identify commercially available microscope-free microsurgical training models or kits not reported in peer-reviewed literature. Literature search identified 1,805 publications; 24 of these met inclusion criteria. Magnification tools most commonly included smartphones (n = 10), VR simulators (n = 4), and tablets (n = 3), with magnification ranging up to ×250 magnification on digital microscopy, ×50 on smartphones, and ×5 on tablets. Average cost of training models ranged from $13 (magnification lens) to $15,000 (augmented reality model). Model were formally assessed using workshops with trainees or attendings (n = 10), surveys to end-users (n = 5), and single-user training (n = 4); users-reported satisfaction with training models and demonstrated faster suturing speed and increased suturing quality with model training. Five commercially available microsurgery training models were identified through Google search. Benchtop microsurgery trainers using digital magnification successfully provide trainees with increased ease of microsurgery training. Low-cost yet high magnification setups using digital microscopes and smartphones are optimal for trainees to improve microsurgical skills. Our assembly protocol, "1, 2, 3, Microsurgery," provides instructions for training model set up to fit the unique needs of any microsurgery trainee.


High detection rate of circulating tumor cells in blood of patients with prostate cancer using telomerase activity.
Circulating tumor cells (CTCs) cannot be readily detected with currently available methods in the majority of patients with prostate cancer. Telomerase activation, one of the major immortalization events, is found in most cases of prostate cancer. We attempted to develop a method using telomerase activity to isolate CTCs in patients with prostate cancer. Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood using Ficoll-Hypaque. Immunomagnetic beads coated with an epithelial cell-specific antigen antibody (BerEP4) were used to harvest epithelial cells from PBMCs. Telomerase activity was detected in harvested epithelial cells using the telomerase-PCR-enzyme-linked immunosorbent assay method. Blood samples from 107 patients with prostate cancer were studied. CTCs were detected in 19 of 24 (79%) patients with advanced prostate cancer. In contrast, CTCs were not detected in blood samples from 22 healthy male volunteers. CTCs were even identified in patients with an undetectable (<0.1 ng/ml) serum prostate-specific antigen (PSA). CTCs were detected in 55 of 70 (79%) patients with localized prostate cancer before radical prostatectomy (n = 30) or brachytherapy (n = 40). CTCs were also detected in 3 of 13 patients (23%) with an undetectable serum PSA measured at least 1 year after radical prostatectomy, which is consistent with the expected relapse rate in this setting. CTCs can be detected using telomerase activity in a large majority and a wide variety of patients with prostate cancer, including those with localized disease.


Groupings of Persons With Traumatic Brain Injury: A New Approach to Classifying Traumatic Brain Injury in the Post-Acute Period.
To (1) identify groups of persons with traumatic brain injury (TBI) who differ on 12 dimensions of cognitive function: cognitive, emotional, and physical symptoms; personal strengths; physical functioning; environmental supports; and performance validity; and (2) describe patterns of differences among the groups on these dimensions and on participation outcome. Three centers for rehabilitation of persons with TBI. A total of 504 persons with TBI living in the community who were an average (standard deviation) of 6.3 (6.8) years postinjury and who had capacity to give consent, could be interviewed and tested in English, and were able to participate in an assessment lasting up to 4 hours. Observational study of a convenience sample of persons with TBI. Selected scales from the Traumatic Brain Injury Quality of Life measures, Neurobehavioral Symptom Inventory, Economic Quality of Life Scale, Family Assessment Device General Functioning Scale, measures of cognitive function, Word Memory Test, and Participation Assessment with Recombined Tools-Objective (PART-O) scale. Cluster analysis identified 5 groups of persons with TBI who differed in clinically meaningful ways on the 12 dimension scores and the PART-O scale. Cluster groupings identified in this study could assist clinicians with case conceptualization and treatment planning.


Surgery in transsexuals.
A review was made of transsexual patients who presented at a clinic for treatment. A total of 154 patients were seen over 6 years. There were 16 female to male transsexuals and 138 male to female transsexuals. Psychiatric screening was a mandatory prerequisite. Hormonal treatment for the transsexual had been started with no exception, prior to consultation. The scope of surgery and its limitations were described to the patient. The female to male transsexual had to undergo a multistaged procedure that included mastectomy, hysterectomy, raising of the penile pedicle, mobilising of the pedicle, insertion of silicone stiffener, and use of testicular prosthesis. Urethral diversion was not attempted. The surgical technique for the male transsexual utilised an inverted U flap on the perineum. Bilateral orchidectomy was performed. The penile skin flap was filletted. The vaginal tunnel was created by sharp and blunt dissection. The urethra was mobilised and translocated. The excess scrotal skin formed the labia. The most serious complication that occurred was that of rectovaginal fistula. Other complications included vaginal stenosis, urethral stenosis and labial abscess.


Genetic variation regulates the activation and specificity of Restriction-Modification systems in Neisseria gonorrhoeae.
Restriction-Modification systems (RMS) are one of the main mechanisms of defence against foreign DNA invasion and can have an important role in the regulation of gene expression. The obligate human pathogen Neisseria gonorrhoeae carries one of the highest loads of RMS in its genome; between 13 to 15 of the three main types. Previous work has described their organization in the reference genome FA1090 and has inferred the associated methylated motifs. Here, we studied the structure of RMS and target methylated motifs in 25 gonococcal strains sequenced with Single Molecule Real-Time (SMRT) technology, which provides data on DNA modification. The results showed a variable picture of active RMS in different strains, with phase variation switching the activity of Type III RMS, and both the activity and specificity of a Type I RMS. Interestingly, the Dam methylase was found in place of the NgoAXI endonuclease in two of the strains, despite being previously thought to be absent in the gonococcus. We also identified the real methylation target of NgoAXII as 5'-GCAGA-3', different from that previously described. Results from this work give further insights into the diversity and dynamics of RMS and methylation patterns in N. gonorrhoeae.


Flying under the radar - impact and factors influencing asymptomatic DENV infections.
The clinical outcome of DENV and other Flaviviruses infections represents a spectrum of severity that ranges from mild manifestations to severe disease, which can ultimately lead to death. Nonetheless, most of these infections result in an asymptomatic outcome that may play an important role in the persistent circulation of these viruses. Also, although little is known about the mechanisms that lead to these asymptomatic infections, they are likely the result of a complex interplay between viral and host factors. Specific characteristics of the infecting viral strain, such as its replicating efficiency, coupled with host factors, like gene expression of key molecules involved in the immune response or in the protection against disease, are among crucial factors to study. This review revisits recent data on factors that may contribute to the asymptomatic outcome of the world's widespread DENV, highlighting the importance of silent infections in the transmission of this pathogen and the immune status of the host.


Provision and use of radiotherapy in Europe.
Radiation therapy is one of the core components of multidisciplinary cancer care. Although ~ 50% of all European cancer patients have an indication for radiotherapy at least once in the course of their disease, more than one out of four cancer patients in Europe do not receive the radiotherapy they need. There are multiple reasons for this underutilisation, with limited availability of the necessary resources - in terms of both trained personnel and equipment - being a major underlying cause of suboptimal access to radiotherapy. Moreover, large variations across European countries are observed, not only in available radiotherapy equipment and personnel per inhabitant or per cancer patient requiring radiotherapy, but also in workload. This variation is in part determined by the country's gross national income. Radiation therapy and technology are advancing quickly; hence, recommendations supporting resource planning and investment should reflect this dynamic environment and account for evolving treatment complexity and fractionation schedules. The forecasted increase in cancer incidence, the rapid introduction of innovative cancer treatments and the more active involvement of patients in the healthcare discussion are all factors that should be taken under consideration. In this continuously changing oncology landscape, reliable data on the actual provision and use of radiotherapy, the optimal evidence-based demand and the future needs are crucial to inform cancer care planning and address and overcome the current inequalities in access to radiotherapy in Europe.


Pharmacokinetics and tolerance of florfenicol in Equidae.
Florfenicol was administered to horses and ponies at a dose rate of 22 mg/kg bwt by i.v., i.m. and oral routes. Following i.v. administration it had an elimination half-life of 1.8 ± 0.9 h, a body clearance of 0.4 ± 0.11/h.kg and a volume of distribution at steady-state of 0.7 ± 0.2 1/kg. It was highly bioavailable following i.m. (81%) and oral (83%) administration. Less than 15% of the administered dose was excreted unchanged in the urine during the 30 h following administration. Animals treated with florfenicol had elevated bilirubin concentrations. Florfenicol was well tolerated by animals in the present study although all animals had loose faeces following administration by each route. At present, florfenicol cannot be recommended for clinical use until multiple dose studies have been carried out to confirm its safety.


The role of the pneumatic antishock garment in penetrating cardiac wounds.
OBJECTIVE--To determine the impact of the pneumatic antishock garment on survival in patients with cardiac wounds. DESIGN--A retrospective review of a 6.5-year experience in a population of patients with gunshot or stab wounds to the heart (N = 70). SETTING--The city and county of Denver, Colo, where all patients were transported by paramedic ambulances to Denver General Hospital, an urban, level 1 trauma center. PATIENTS--All patients were analyzed, including those transported to the emergency department with no vital signs at the trauma scene. Only patients with definite penetrating cardiac injuries were included. METHODS--Paramedic trip sheets and emergency department, operating room, in-hospital, and autopsy records were reviewed. Demographic, clinical outcome, and treatment data were collected. A multiple logistic regression model was developed to determine which characteristics and treatment variables were independently predictive of patient outcome. The ability of each independent variable to predict outcome was determined by calculating adjusted odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS--Seventy patients (31 gunshot and 39 stab wounds) had a revised trauma score of 2.8 +/- 4.0 (mean +/- SD; range, 0-12; median, 0). Thirty-two (46%) had cardiac tamponade. Overall, 21 (30%) of the patients survived. In the logistic model, the presence of cardiac tamponade (OR, 9.1; 95% CI, 1.5 to 56.1) and a higher revised trauma score were associated with higher survival. Inflation of the pneumatic antishock garment was associated with sevenfold lower survival (adjusted OR, 0.14; 95% CI, 0.02 to 0.86); the effect was negative across all trauma score groups. --Survival in patients with penetrating cardiac wounds is highly associated with anatomic and physiologic factors, while application of the pneumatic antishock garment appears to be detrimental.


Formation and origin of multicenter photoluminescence in zeolite-based carbogenic nanodots.
Carbogenic nanodots (CNDs) are inspiring intensive research efforts owing to their excellent optical properties; however, their chemical structures and photoluminescent origins are still under study. Herein, CNDs with interesting multicenter photoluminescence (PL) have been successfully prepared by pyrolysis of organo-templated zeolites. The simultaneously observed excitation-dependent blue and excitation-independent green PL emissions as well as an uncommon ultraviolet-excited cyan emission are quite different from those of the known multicenter emissive CNDs. To get a thorough understanding of the formation of these CNDs and their PL origins, structural characterizations coupled with femtosecond transient absorption and fluorescence lifetime spectra analyses have been employed to study such PL behavior. The results show that the as-prepared CNDs consist of three main batches with different polarities, and they are composed of a combination of six small fluorophores with diverse ratios. The multicenter PL is obtained due to competition between the conjugated domain fluorophores and the surface-occupied functional group fluorophores. This study provides a comprehensive insight into the origin of multicenter photoluminescence in zeolite-based CNDs.


Relationship between measures of aerobic fitness, speed and repeated sprint ability in full and part time youth soccer players.
The aim of the study was to investigate the relationship between repeated sprint ability (RSA) involving changes in direction, short linear sprinting and aerobic capacity in young elite soccer players. A secondary aim was to assess any differences in performance of these assessments between players of different age groups. Thirty-two male adolescent soccer players belonging to the same elite club academy were assessed for RSA comprising 6 x 40m efforts interspersed by 25s recovery, linear sprinting speed over 15m, and aerobic capacity via the YYIE2 assessment. There was a significant correlation between performance in the YYIE2 and RSA total time, RSA fastest sprint and RSA percentage decrement (r = -0.71, -0.53, and -0.52 respectively. P<0.05). Significant differences were observed between age groups for distance covered in the YYIE2, RSA total time and RSA fastest sprint (P<0.01). No significant differences were observed for short linear sprinting. Assessments of RSA over 40m and incorporating changes of direction appear to be significantly correlated with YYIE2 performance in young elite level soccer players. In addition older players performed significantly better in the YYIE2 assessment and RSA protocol but not in short linear sprinting. These results have implications for the design of assessment protocols for young elite soccer players of different ages.


[MURCS association: case report].
Clinical term of association refers to a not randomized congenital malformations which are present in one single subject. The term MURCS is an acronym for (MU) Mullerian, (R) Renal, (C) Cervicothoracic, (S) Somite abnormalities. We communicate a case of a phenotipically normal 16 years old female patient with primary amenorrhea due to müllerian malformations and cervicothoracic dysplasia integrating the MURCS association diagnosis.


Aspergillus terreus spondylodiscitis following an abdominal stab wound: a case report.
Aspergillus terreus, a saprophytic fungus, is recognized as an emerging pathogen in various infections in humans. However, bone and joint involvement is uncommon. To the best of our knowledge, only seven cases of spondylodiscitis caused by Aspergillus terreus have been reported previously in humans. We report a case of a patient with Aspergillus terreus spondylodiscitis following an abdominal stab wound. A 74-year-old Japanese man with no particular medical history fell from a ladder and sustained a left abdominal stab wound from an L-shaped metal peg. Computed tomography showed the trace of the L-shaped metal peg from the left abdomen to the left rib and left kidney. The scan also showed an anterolateral bone avulsion of the left side of the T12 vertebral body, as well as fractures of the L1 left transverse process and the left 10th-12th ribs. He was hospitalized and treated with conservative therapy for 6 weeks. He was readmitted to the hospital with complaints of sudden back pain, numbness of both legs, and inability to walk 13 weeks after the fall. Magnetic resonance imaging findings were typical of spondylodiscitis. Gadolinium-enhanced T1-weighted magnetic resonance imaging indicated increased signal intensity at T11-T12 vertebral bodies and severe cord compression and epidural abscess at T11-T12 associated with infiltration of soft paravertebral tissues. On the seventh day after admission, he underwent partial laminectomy at T11 and posterior fusion at T9 to L2. The result of his blood culture was negative, but Aspergillus terreus was isolated from the material of T11-T12 intervertebral disc and vertebral bodies. His Aspergillus antigen was positive in a blood examination. Histological examination showed chronic suppurative osteomyelitis. On the 35th day after admission, he underwent anterior fusion at T11 and T12 with a rib bone graft. For 5 months, voriconazole was administered, and he wore a rigid corset. Posterior partial laminectomy at T11 and anterior fusion at T11 and T12 resulted in a good clinical course. The patient's neurological dysfunction was completely recovered, and his back pain disappeared. Two years after the operation, computed tomography was performed and showed bone fusion at T11 and T12. Magnetic resonance imaging revealed no evidence of increased signal intensity at T11-T12 vertebral bodies and severe cord compression and epidural abscess at T11-T12. To our knowledge, this is the first report of spondylodiscitis caused by Aspergillus terreus after an abdominal penetrating injury. The histological finding of chronic suppurative osteomyelitis and the radiological findings strongly suggested direct inoculation of Aspergillus terreus.


Raised intracellular glucose concentrations reduce aggregation and cell death caused by mutant huntingtin exon 1 by decreasing mTOR phosphorylation and inducing autophagy.
Huntington's disease is caused by a CAG trinucleotide repeat expansion that is translated into an abnormally long polyglutamine tract. This gain-of-function mutation is associated with huntingtin aggregation and cell death. Autophagy is an important clearance route for mutant huntingtin exon 1. While mammalian target of rapamycin (mTOR) is a key regulator of autophagy, the upstream modifiers of this process are poorly understood. Our previous expression profiling studies in HD cell models observed changes in four genes associated with glucose metabolism, including the GLUT1 glucose transporter. A role for intracellular glucose as a modulator for polyglutamine toxicity was suggested as cell death was reduced by GLUT1 overexpression. Here we show that the protective effect of GLUT1 is associated with decreased huntingtin exon 1 aggregation in cell models. Consistent with this result, we also observed reduced aggregation and enhanced clearance of mutant huntingtin when cells were cultured in raised glucose concentrations (8 g/l). These effects were mimicked by 8 g/l 2-deoxyglucose (2DOG) (transported, phosphorylated but not metabolized further), but not with 8 g/l 3-O-methyl glucose (transported but not metabolized further). Thus, this phenomenon is probably mediated by glucose-6-phosphate. Increased clearance of mutant huntingtin by raised glucose (8 g/l) and 2DOG correlated with increased autophagy and reduced phosphorylation of mTOR, S6K1 and Akt. Thus, raised intracellular glucose/glucose 6-phosphate levels reduce mutant huntingtin toxicity by increasing autophagy via mTOR and possibly Akt. As mTOR and Akt regulate a diversity of crucial cellular processes, our data also suggest a major new set of targets for intracellular glucose signalling.


Pathways of fear and anxiety in endodontic patients.
To evaluate the most common pathways of fear and anxiety in patients who have had root canal treatment or are planned to have one. Six hundred and twenty-seven patients were approached to participate, of which five hundred and ninety-four patients (20-90 years) accepted. All consenting patients had a root filling or were treatment planned to have one. The survey by Ost & Hugdahl () on anxiety response patterns was modified and used. Data were presented using descriptive statistics, tested for normality using the Kolmogorov-Smirnov test and analysed with nonparametric anova (Kruskal-Wallis) and post hoc test. Cognitive conditioning and parental pathways seem to be the primary cause (P < 0.05) of fear and anxiety with root canal treatment. Females were significantly more likely to be influenced by indirect conditioned experiences such as informative, parental, verbal threat and vicarious pathways. The origin of patients' fears requires more attention in terms of treating endodontic-related fear and anxiety. More detailed research into the effects of demographics, causative factors and ethnicity on pathways of fear in dentistry is required to help dentists better manage patients in a multicultural society.


Infection and chronic disease activate a brain-muscle signaling axis that regulates muscle performance.
Infections and neurodegenerative diseases induce neuroinflammation, but affected individuals often show a number of non-neural symptoms including muscle pain and muscle fatigue. The molecular pathways by which neuroinflammation causes pathologies outside the central nervous system (CNS) are poorly understood, so we developed three models to investigate the impact of neuroinflammation on muscle performance. We found that bacterial infection, COVID-like viral infection, and expression of a neurotoxic protein associated with Alzheimer' s disease promoted the accumulation of reactive oxygen species (ROS) in the brain. Excessive ROS induces the expression of the cytokine Unpaired 3 (Upd3) in insects, or its orthologue IL-6 in mammals, and CNS-derived Upd3/IL-6 activates the JAK/Stat pathway in skeletal muscle. In response to JAK/Stat signaling, mitochondrial function is impaired and muscle performance is reduced. Our work uncovers a brain-muscle signaling axis in which infections and chronic diseases induce cytokine-dependent changes in muscle performance, suggesting IL-6 could be a therapeutic target to treat muscle weakness caused by neuroinflammation.


Transradial coronary angioplasty and stenting--immediate results and 3-month clinical follow-up in the first 50 patients performed at the National Heart Institute.
To explore the safety and feasibility of coronary angioplasty and stenting via the radial artery in a heterogenous group of patients and to report the immediate and 3-month clinical follow-up. The use of the transradial approach for coronary angiography was first described by Lucien Campeau in 1989. Based on the favourable initial results, this technique has gained widespread acceptance worldwide. Ferdinand Kiemeneij's work in transradial angioplasty and stenting has taken invasive cardiology into the exciting new era of "minimally invasive coronary intervention". Fifty consecutive patients underwent Transradial Percutaneous Transluminal Coronary Angioplasty (PTCA) with or without stenting from mid March 98-December 98. The right radial approach was utilised in 41 patients (80%) while the left in 9 patients. Ninety percent of the procedures was done on an adhoc basis. Diabetes mellitus was present in 38% of patients. Eighty percent of patients had unstable angina pectoris and 60% had a prior history of acute myocardial infarction. The commonest vessel involved was the LAD (51%) and type B lesions predominated (54%). PTCA was successful in 96%. One patient had a total LAD occlusion, which could not be wired, and another developed severe spasm during catheter manipulation. The latter had successful PTCA via the right femoral route Stents were utilised in 57% of patients. The commonest indication for stenting was suboptimal PTCA results (89%) and dissection (14%). There was no stent embolisation and all stents were successfully deployed (100%). One patient developed acute stent thrombosis necessitating repeat PTCA and another patient sustained an acute anteroseptal myocardial infarction 5 days post procedure as a result of subacute stent thrombosis and died. Two patients had successful primary PTCA. There was no bleeding or vascular complications. 60% of patients were treated on an outpatient basis. At 3-months follow up, 1 patient required CABG's for disease progression. Three patients had absent radial pulses without adverse consequence. No patient required repeat PTCA at follow up. In summary, adhoc PTCA and stenting is safe and feasible in our patient population. A study on the cost effectiveness of the procedure compared to conventional femoral PTCA is warranted.


The anti-apoptotic Bcl-2 family member Mcl-1 promotes T lymphocyte survival at multiple stages.
T lymphocyte development and function are tightly regulated by the intrinsic death pathway through members of the Bcl-2 family. Genetic studies have demonstrated that the Bcl-2 family member Mcl-1 is an important anti-apoptotic protein in the development of multiple cell types including T lymphocytes. However, the expression pattern and anti-apoptotic roles of Mcl-1 in T lymphocytes at different developmental stages remain to be fully determined. In this study, we examined the expression pattern of Mcl-1 in different populations of T cells at the single-cell level and found that Mcl-1 protein is constitutively expressed in all T cell populations and up-regulated upon TCR stimulation. We then investigated the role of Mcl-1 in the survival of these different populations by conditionally deleting Mcl-1 at various T cell stages. Our results show that Mcl-1 is required for the survival of double-negative and single-positive thymocytes as well as naive and activated T cells. Furthermore, we demonstrate that Mcl-1 functions together with Bcl-xL to promote double-positive thymocyte survival. Thus, Mcl-1 is a critical anti-apoptotic factor for the survival of T cells at multiple stages in vivo.


[Effects of chronic buspirone treatment on female mice exposed to the long-lasting psychoemotional influence].
The effects of chronic treatment (30 days) with the 5-HT1A receptor partial agonist buspirone (0.05, 1 and 10 mg/kg i.p.) on the behaviour of C57BL/6J female mice exposed to long-lasting psychoemotional influence were studied. The influence involved forced living of each female with an aggressive male separated with a perforated transparent partition in the same cage and daily female's presence during 10-min intermale confrontations behind a partition caused by introducing of another male to the aggressive male. Chronic buspirone injection (in all used doses) did not affect the behaviour of females estimated in the "partitions" and "open field" tests at the end of the drug treatment. The anxiolytic effect of buspirone only at the dose of 1 mg/kg on the female's plus-maze behaviour was revealed. In the Porsolt, test buspirone in the dose of 1 mg/kg caused a slight increase in the duration of immobility indicating a slight pro-depressive effect. Thus, chronic buspirone treatment of females exposed to the long-lasting psychoemotional impact has a different effect on their behaviour depending on the dose and test conditions.


A free-fall determination of the newtonian constant of gravity
Recent determinations of the Newtonian constant of gravity have produced values that differ by nearly 40 times their individual error estimates (more than 0.5%). In an attempt to help resolve this situation, an experiment that uses the gravity field of a one-half metric ton source mass to perturb the trajectory of a free-falling mass and laser interferometry to track the falling object was performed. This experiment does not suspend the test mass from a support system. It is therefore free of many systematic errors associated with supports. The measured value was G = (6.6873 +/- 0. 0094) x 10(-11) m3 kg-1 sec-2.


Cloning and sequencing of the Escherichia coli chlEN operon involved in molybdopterin biosynthesis.
The nucleotide sequence of a HinPI-HpaII restriction nuclease fragment which complemented a delta chlE strain of Escherichia coli was determined. Two open reading frames were deduced to be the structural genes for ChlE and ChlN proteins, which have molecular weights of 44,067 and 26,719, respectively. Both proteins were required for complementing a chromosomal deletion of the chlE locus. The chlE and chlN genes were transcribed from a common promoter, chlEp, located upstream of chlE. Transcriptional and translational signal sequences were recognized in this region.