Catalase evolved to concentrate H2O2 at its active site.
Catalase is a homo-tetrameric enzyme that has its heme active site deeply buried inside the protein. Its only substrate, hydrogen peroxide (H2O2), reaches the heme through a 45 A-long channel. Large-subunit catalases, but not small-subunit catalases, have a loop (gate loop) that interrupts the major channel. Two accesses lead to a gate that opens the final section of the channel to the heme; gates from the R-related subunits are interconnected. Using molecular dynamic simulations of the Neurospora crassa catalase-1 tetramer in a box of water (48,600 molecules) or 6M H2O2, it is shown that the number of H2O2 molecules augments at the surface of the protein and in the accesses to the gate and the final section of the channel. Increase in H2O2 is due to the prevalence and distribution of amino acids that have an increased residency for H2O2 (mainly histidine, proline and charged residues), which are localized at the protein surface and the accesses to the gate. In the section of the channel from the heme to the gate, turnover rate of water molecules was faster than for H2O2 and increased residence sites for water and H2O2 were determined. In the presence of H2O2, the exclusion of water molecules from a specific site suggests a mechanism that could contend with the competing activity of water, allowing for catalase high kinetic efficiency.


Contributions by members of the TGFbeta superfamily to lens development.
Members of the TGFbeta superfamily of growth and differentiation factors, including the TGFbeta, BMP, activin and nodal families, play important signaling roles throughout development. This paper summarizes some of the functions of these ligands in lens development. Targeted deletion of the genes encoding one of the BMP receptors, Alk3 (BMP receptor-1A), showed that signaling through this receptor is essential for normal lens development. Lenses lacking Alk3 were smaller than normal, with thin epithelial layers. The fiber cells of Alk3 null lenses became vacuolated and degenerated within the first week after birth. Lenses lacking Alk3 function were surrounded by abnormal mesenchymal cells, suggesting that the lenses provided inappropriate signals to surrounding tissues. Lens epithelial and fiber cells contained endosomes that were associated with activated (phosphorylated) SMAD1 and SMAD2. Endosomal localization of pSMAD1 was reduced in the absence of Alk3 signaling. The presence of pSMAD2 in lens fiber cell nuclei and the observation that the activin antagonist follistatin inhibited lens cell elongation suggested that an activin-like molecule participates in lens fiber cell differentiation. Lenses deficient in type II TGFbeta receptors were clear and had fiber cells of normal morphology. This suggests that TGFbeta signaling is not essential for the normal differentiation of lens fiber cells. The targeted deletion of single or multiple receptors of the TGFbeta superfamily in the lens should further characterize the role of these signaling molecules in lens development. This approach may also provide a useful way to define the downstream pathways that are activated by these receptors during the development of the lens and other tissues.


Venous thrombosis and procoagulant factors in high-risk neuroblastoma.
The mechanism of increased thrombin production has been investigated in children with high-risk neuroblastoma (NB), to detect any possible association between catheter- related venous thrombosis (VT) and prothrombotic factors. Consecutive children with high-risk NB were studied by color-doppler ultrasonography of the upper vein system and thrombophilia factors assessment. Plasma levels of Tissue Factor (TF), Vascular Endothelial Growth Factor (VEGF), Prothrombin Activation Fragment 1+2, and Thrombin-Antithrombin Complex were evaluated. Moreover, inherited thrombophilia factors (homocystein, antithrombin, protein C, protein S, factor V Leiden, activated protein C resistence, mutation H1299R and G1691A of factor V, mutation G20210A of prothrombin, mutation T677 and A1298C of methylenetetrahydrofolate reductase, and allele 4G of plasminogen activator inhibitor-1) were tested to exclude congenital disorders. Six patients with mean age: 48.8 months---were studied. Five patients were affected by stage 4 NB and another one by stage 3 NB with Myc-N amplification. All children had a central venous line (mean duration: 8.5 mos). Four patients (67%) had asymptomatic catheter-related VT visualized by color-doppler ultrasonography. No patient had major inherited thrombophilia factors. The levels of plasma TF and plasma VEGF were found elevated in all patients. Mean value of TF (nv 20.3+/-6.6) was 82 pg/mL with a range of 39 to 131 pg/mL. Mean value of VEGF (nv 24.3 pg/mL) was 78.5 pg/mL with a range of 31 to 142 pg/mL. The increased risk of catheter-related VT detected in our small series of high-risk NB patients, was associated with elevated levels of circulating TF and VEGF. Further studies are needed to evaluate if elevated levels of TF/VEGF are involved both in the hypercoagulable state and in advanced childhood cancer.


Update on insulin therapy for type 2 diabetes.
Type 2 diabetes is characterized by insulin resistance and impaired insulin secretion at diagnosis and by progressive β-cell dysfunction over time. Insulin therapy is thus frequently required during the course of the disease to maintain glycemic control and prevent diabetes complications. Insulin should be initiated when alternative antihyperglycemic agents have failed or when symptomatic or marked hyperglycemia is present. Recent studies demonstrate that the addition of basal, prandial, basal/bolus, or premixed insulins to existing antihyperglycemic regimens effectively lowers glycosylated hemoglobin (HbA(1c)). The long-acting insulin analogs cause less nocturnal hypoglycemia than bedtime NPH, with comparable HbA(1c) reductions. Insulin detemir confers a weight advantage over glargine or NPH. Rapid-acting insulin analogs control postprandial hyperglycemia more effectively than regular insulin and modestly lower HbA(1c). For selected patients with severe insulin resistance, U-500 is a less expensive and potentially more effective alternative to U-100 insulin. Adverse effects of insulin, including weight gain and hypoglycemia, can be minimized by initial use of basal insulins in combination with metformin, incretin mimetics, or dipeptidyl-peptidase-IV inhibitors. Although in vitro studies suggest that hyperinsulinemia may promote tumorigenesis, no currently available insulin has been shown to increase cancer rates. Targeting near-normal glucose levels in insulin-treated patients should be reserved for those of younger age with a longer life expectancy, a shorter duration of diabetes, and little or no end-organ complications. A higher HbA(1c) target of 7-8% is more appropriate for patients less likely to benefit from intensive control and in those at high risk for severe hypoglycemia.


Light affects ascorbate content and ascorbate-related gene expression in tomato leaves more than in fruits.
Little is known about the light regulation of vitamin C synthesis in fruits. In contrast, previous studies in leaves revealed that VTC2 (coding for GDP-L: -galactose phosphorylase) was one of the key genes up-regulated by light in leaves. Our objective was to determine how the expression of ascorbate (AsA) synthesis genes in tomato (Solanum lycopersicum) was modified according to light irradiance in both leaves and fruits. Seven days of shading strongly decreased total ascorbate (reduced and oxidized form) content in leaves (50%) and to a lesser extent in fruits (10%). Among the last six steps of AsA biosynthesis, only two genes, VTC2 and GPP1 (one of the two unigenes coding for L: -galactose-1-P phosphatase in tomato), were down-regulated by long-term shading in red ripe fruits, compared to seven genes regulated in leaves. This underlines that light affects AsA-related gene expression more in leaves than in ripening fruits. Moreover, this study reveals strong daily changes in transcript levels of enzymes of the AsA biosynthetic pathway in leaves (11 of the 12 studied genes showed significant changes in their expression pattern). Among those genes, we found that diurnal variation in transcript levels of VTC2 and GME1 correlated to leaf AsA content measured 8 h later. This study provides a new hypothesis on the role of GME1 in addition to VTC2 in light-regulated AsA biosynthesis.


Coding of the contrasts in natural images by visual cortex (V1) neurons: a Bayesian approach.
Individual V1 neurons respond dynamically over only limited ranges of stimulus contrasts, yet we can discriminate contrasts over a wide range. Different V1 neurons cover different parts of the contrast range, and the information they provide must be pooled somehow. We describe a probabilistic pooling model that shows that populations of neurons with contrast responses like those in cat and monkey V1 would most accurately code contrasts in the range actually found in natural scenes. The pooling equation is similar to Bayes's equation; however, explicit inclusion of prior probabilities in the inference increases coding accuracy only slightly.


Statin therapy helps to control blood pressure levels in hypertensive dyslipidemic patients.
Aside from lowering lipid levels; statins improve endothelial function, decrease platelet aggregation, reduce procoagulant blood factors, and decrease vascular tone. This study was conducted to investigate the possible effect of atorvastatin on blood pressure (BP) in a group of hypertensive and dyslipidemic patients. Thirty-six hypertensive and dyslipidemic patients with inadequately controlled lipid levels by diet were treated with atorvastatin 20 mg/day for 8 weeks and compared with 24-patient matched controls treated with diet alone. The type and dosage of antihypertensive medications were not altered during statin therapy. Blood lipid profile including total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL) and triglyceride (TG) levels were noted at inclusion and after 8 weeks. Ambulatory BP monitoring (ABPM) was carried out at study entry and at the end of week eight. A total of 49 patients (32 patients in the atorvastatin group and 17 patients in the control group) completed the 3-month follow-up period of observation. The ABPM studies indicated significant reductions in total average systolic BP, total average diastolic BP, total average mean BP, day average systolic BP, day average diastolic BP, night average systolic BP, night average diastolic BP, and night average mean BP levels in the atorvastatin group, whereas these reductions were not observed in the control group. Our results indicate that atorvastatin therapy significantly improves BP control in hyperlipidemic hypertensive patients. However, the effects of other statins on BP, as well as, the different dosages need to be further investigated.


Applications of multi-nuclear magnetic resonance spectroscopy at 7T.
To discuss the advantages of ultra-high field (7T) for (1)H and (13)C magnetic resonance spectroscopy (MRS) studies of metabolism. Measurements of brain metabolites were made at both 3 and 7T using (1)H MRS. Measurements of glycogen and lipids in muscle were measured using (13)C and (1)H MRS respectively. In the brain, increased signal-to-noise ratio (SNR) and dispersion allows spectral separation of the amino-acids glutamate, glutamine and γ-aminobutyric acid (GABA), without the need for sophisticated editing sequences. Improved quantification of these metabolites is demonstrated at 7T relative to 3T. SNR was 36% higher, and measurement repeatability (% coefficients of variation) was 4%, 10% and 10% at 7T, vs 8%, 29% and 21% at 3T for glutamate, glutamine and GABA respectively. Measurements at 7T were used to compare metabolite levels in the anterior cingulate cortex (ACC) and insula. Creatine and glutamate levels were found to be significantly higher in the insula compared to the ACC (P < 0.05). In muscle, the increased SNR and spectral resolution at 7T enables interleaved studies of glycogen ((13)C) and intra-myocellular lipid (IMCL) and extra-myocellular lipid (EMCL) ((1)H) following exercise and re-feeding. Glycogen levels were significantly decreased following exercise (-28% at 50% VO(2) max; -58% at 75% VO(2) max). Interestingly, levels of glycogen in the hamstrings followed those in the quadriceps, despite reduce exercise loading. No changes in IMCL and EMCL were found in the study. The demonstrated improvements in brain and muscle MRS measurements at 7T will increase the potential for use in investigating human metabolism and changes due to pathologies.


The Australian Twin Study of Gambling (OZ-GAM): rationale, sample description, predictors of participation, and a first look at sources of individual differences in gambling involvement.
Two major challenges to conducting a community-based twin study of pathological gambling (PG) disorder are that: (a) it is relatively rare, and (b) individuals with the disorder in the community may be difficult to locate and recruit. We describe a new study of 4,764 individuals recruited from the Australian Twin Registry in which we attempt to effectively deal with the first challenge and examine the impact of the second challenge. The lifetime prevalence of DSM-IV PG in this Australian twin sample was 2.2%, which is 400-500% higher than has been obtained in prevalence surveys conducted in the United States. A number of predictors of non-participation were identified, including a lifetime PG disorder diagnosis, but these did not have a large net effect on the estimated number of individuals with PG or related characteristics in the twin sample. Results of biometric modeling suggested that the effect of genetic, shared family environmental, and nonshared environmental influences on the propensity to engage in 11 different specific forms of gambling (e.g., playing the lottery, betting on horse or dog races, playing electronic gaming machines) were generally moderate, low, and moderate, respectively, with mean parameter estimates obtained of 43%, 10%, and 46%. An intriguing comparison with results from a 1963 US adolescent twin study conducted by Loehlin and Nichols (1976) suggests that: (a) propensity genes for gambling involvement may be more likely to be expressed in the heavy-gambling Australian culture, or that (b) the family environment has a transient effect on the gambling behavior of young people.


Use of contemporary biomarkers in the detection of chronic alcohol use.
Alcohol is the most commonly abused substance yet alcoholism is frequently undiagnosed. The misuse of alcohol is common and frequently an occult problem. More than 10% of current drinkers meet diagnostic criteria for alcohol abuse or dependence while the lifetime prevalence for these conditions in outpatient settings ranges from 16 to 36 percent. Long-term, heavy drinking is associated with significant morbidity, mortality, and economic costs. Clues to alcohol use can be discovered from a patient's history and physical stigmata. Validated screening instruments such as the Alcohol Use Disorders Identification Test (AUDIT), CAGE Questionnaire, and Brief Michigan Alcoholism Screening Tests help confirm the clinical suspicion of alcohol dependence. Laboratory abnormalities of mean corpuscular volume, gamma-glutamyl transferase, alkaline phosphatase, or alanine amino transferase levels are non-specific indicators of possible alcohol-induced liver impairment. Newer, less well-known FDA-approved biochemical markers such as the Carbohydrate Deficient Transferrin and the Early Detection of Alcohol Consumption test may also be used to detect heavy alcohol abuse and to monitor relapse episodes. Brief interventions are successful, making identification and diagnosis a vital role for the family physician. Improved awareness of alcohol misuse, increased use of screening tools, and the appropriate use of biochemical markers will facilitate early intervention and successful management of patients with alcohol use disorders.


Sublimation-based wafer-scale monolayer WS2 formation via self-limited thinning of few-layer WS2.
Atomically-thin monolayer WS2 is a promising channel material for next-generation Moore's nanoelectronics owing to its high theoretical room temperature electron mobility and immunity to short channel effect. The high photoluminescence (PL) quantum yield of the monolayer WS2 also makes it highly promising for future high-performance optoelectronics. However, the difficulty in strictly growing monolayer WS2, due to its non-self-limiting growth mechanism, may hinder its industrial development because of the uncontrollable growth kinetics in attaining the high uniformity in thickness and property on the wafer-scale. In this study, we report a scalable process to achieve a 4 inch wafer-scale fully-covered strictly monolayer WS2 by applying the in situ self-limited thinning of multilayer WS2 formed by sulfurization of WOx films. Through a pulsed supply of sulfur precursor vapor under a continuous H2 flow, the self-limited thinning process can effectively trim down the overgrown multilayer WS2 to the monolayer limit without damaging the remaining bottom WS2 monolayer. Density functional theory (DFT) calculations reveal that the self-limited thinning arises from the thermodynamic instability of the WS2 top layers as opposed to a stable bottom monolayer WS2 on sapphire above a vacuum sublimation temperature of WS2. The self-limited thinning approach overcomes the intrinsic limitation of conventional vapor-based growth methods in preventing the 2nd layer WS2 domain nucleation/growth. It also offers additional advantages, such as scalability, simplicity, and possibility for batch processing, thus opening up a new avenue to develop a manufacturing-viable growth technology for the preparation of a strictly-monolayer WS2 on the wafer-scale.


Trends in hazardous trace metal concentrations in aerosols collected in Beijing, China from 2001 to 2006.
Daily observations of hazardous trace metal concentrations in aerosols in Beijing, China were made in the period from 2001 to 2006. We considered coal combustion as a major source of some anthropogenic metals by achieving a correlation analysis and by investigating enrichment factors and relative composition of metals. A possible extra source of some specific metals, such as Cu and Sb, was brake abrasion particles, however, we did not think the transport-related particle was a major source for the hazardous anthropogenic metals even though they could originate from vehicle exhaust and brake/tire abrasion particles. A time-trend model was used to describe temporal variations of chemical constituent concentrations during the five-year period. Several crustal elements, such as Al, Ti, V, Cr, Mn, Fe, and Co, did not show clear increases, with annual rates of change of -15.2% to 3.6%. On the other hand, serious increasing trends were noted from several hazardous trace metals. Cu, Zn, As, Cd, and Pb, which are derived mainly from anthropogenic sources, such as coal combustion, showed higher annual rate of change (4.9-19.8%, p<0.001) according to the regression model. In particular, the Cd and Pb concentrations increased remarkably. We hypothesize that the trend towards increasing concentrations of metals in the air reflects a change that has occurred in the process of burning coal, whereby the use of higher temperatures for coal combustion has resulted in increased emissions of these metals. The increasing use of low-rank coal may also explain the observed trends. In addition, nonferrous metal smelters are considered as a potential, albeit minor, reason for the increasing atmospheric concentrations of anthropogenic hazardous metals in Beijing city.


The role of PET/CT in assessing pulmonary nodules in children with solid malignancies.
OBJECTIVE. The purpose of this article is to assess the feasibility and utility of PET/CT in distinguishing benign from malignant pulmonary nodules in patients with solid childhood malignancies. SUBJECTS AND METHODS. This prospective study was conducted between March 2008 and August 2010. We enrolled 25 subjects 21 years old or younger with solid childhood malignancies and at least one pulmonary nodule measuring 0.5-3.0 cm. PET/CT was performed within 3 weeks of diagnostic chest CT. Three panels of three reviewers each reviewed diagnostic CT only (panel 1), PET/CT only (panel 2), or diagnostic CT and PET/CT concurrently (panel 3) and predicted each nodule's histologic diagnosis as benign, malignant, or indeterminate. Interreviewer agreement was assessed with the kappa statistic. Using nodule biopsy or clinical follow-up as reference standards, the sensitivity, specificity, and accuracy for each panel was assessed. Logistic regression was used to assess the nodule's maximum standardized uptake value (SUVmax) association with its histologic diagnosis. RESULTS. There were 75 nodules with a median size of 0.74 cm (range, 0.18-2.38 cm); 48 nodules were malignant. Sensitivity was 85% (41/48) for panel 1, 60% (29/48) for panel 2, and 67% (32/48) for panel 3. All panels had poor specificities. Interreviewer agreement was moderate for panel 1 (0.43) and poor for panels 2 (0.22) and 3 (0.33). SUVmax was a significant predictor of histologic diagnosis (p = 0.004). CONCLUSION. PET/CT assessment of pulmonary nodules is feasible in children with solid malignancies but may not reliably improve our ability to predict a nodule's histologic diagnosis. The SUVmax may improve the performance of PET/CT in this setting.


Systems Analysis of Early Host Gene Expression Provides Clues for Transient Mycobacterium avium ssp avium vs. Persistent Mycobacterium avium ssp paratuberculosis Intestinal Infections.
It has long been a quest in ruminants to understand how two very similar mycobacterial species, Mycobacterium avium ssp. paratuberculosis (MAP) and Mycobacterium avium ssp. avium (MAA) lead to either a chronic persistent infection or a rapid-transient infection, respectively. Here, we hypothesized that when the host immune response is activated by MAP or MAA, the outcome of the infection depends on the early activation of signaling molecules and host temporal gene expression. To test our hypothesis, ligated jejuno-ileal loops including Peyer's patches in neonatal calves were inoculated with PBS, MAP, or MAA. A temporal analysis of the host transcriptome profile was conducted at several times post-infection (0.5, 1, 2, 4, 8 and 12 hours). When comparing the transcriptional responses of calves infected with the MAA versus MAP, discordant patterns of mucosal expression were clearly evident, and the numbers of unique transcripts altered were moderately less for MAA-infected tissue than were mucosal tissues infected with the MAP. To interpret these complex data, changes in the gene expression were further analyzed by dynamic Bayesian analysis. Bayesian network modeling identified mechanistic genes, gene-to-gene relationships, pathways and Gene Ontologies (GO) biological processes that are involved in specific cell activation during infection. MAP and MAA had significant different pathway perturbation at 0.5 and 12 hours post inoculation. Inverse processes were observed between MAP and MAA response for epithelial cell proliferation, negative regulation of chemotaxis, cell-cell adhesion mediated by integrin and regulation of cytokine-mediated signaling. MAP inoculated tissue had significantly lower expression of phagocytosis receptors such as mannose receptor and complement receptors. This study reveals that perturbation of genes and cellular pathways during MAP infection resulted in host evasion by mucosal membrane barrier weakening to access entry in the ileum, inhibition of Ca signaling associated with decreased phagosome-lysosome fusion as well as phagocytosis inhibition, bias toward Th2 cell immune response accompanied by cell recruitment, cell proliferation and cell differentiation; leading to persistent infection. Contrarily, MAA infection was related to cellular responses associated with activation of molecular pathways that release chemicals and cytokines involved with containment of infection and a strong bias toward Th1 immune response, resulting in a transient infection.


[CBO-guideline for diagnosis and treatment of the acute ankle injury. National organization for quality assurance in hospitals].
Following an acute ankle inversion trauma, the aim of the diagnostic strategy is to rule out a fracture and/or lateral ankle ligament rupture. Plain X-rays are indicated when weightbearing is impossible (the patient cannot take at least four steps) and/or if there is pain on palpation on the posterior aspect of the lateral or medial malleolus. This applies to the period directly following the trauma as well as 4-5 days later. In the first forty-eight hours after trauma, physical examination cannot distinguish between a distortion and a lateral ligament rupture. In a patient with a painful swollen ankle after an acute inversion trauma, a compression bandage is applied and the patient is advised to elevate the leg and to perform ankle movements, notably dorsal extension. Weightbearing on the extremity may be allowed as soon as the pain allows. A few days later, when pain and swelling have subsided, delayed physical examination for detection of a lateral ligament rupture is performed. Delayed physical examination four to five days after inversion trauma has good interobserver agreement. Additional diagnostics such as radiography, arthrography, ultrasound investigation and MRI are expensive and give no further information. In a patient with a simple distortion an elastic bandage for a few days can be applied. The patient is instructed to resume normal walking as soon as possible. Functional treatment with inelastic tape bandage or a brace application for five to six weeks is the treatment of choice for an acute lateral ankle ligament rupture. The aim of treatment is restoration of a normal range of motion within two weeks and a stable ankle at long term follow-up.


Diffusion kurtosis imaging in patients with locally advanced rectal cancer: current status and future perspectives.
Morphological magnetic resonance imaging is currently the best imaging technique for local staging in patients with rectal cancer. However, morphological sequences have some limitations, especially after preoperative chemoradiotherapy (pCRT). Diffusion-weighted imaging has been applied to rectal cancer for detection of lesions, characterization of tissue, and evaluation of the response to therapy. In 2005, a non-Gaussian diffusion model called diffusion kurtosis imaging (DKI) was suggested. Several electronic databases were evaluated in the present review. The search included articles published from January 2000 to May 2018. The references of all articles were also evaluated. All titles and abstracts were assessed, and only the studies of DKI in patients with rectal cancer were retained. We identified 35 potentially relevant references through the electronic search. According to the inclusion and exclusion criteria, we retained five clinical studies that met the inclusion criteria. DKI is a useful tool for assessment of tumor aggressiveness, the nodal status, and the risk of early metastases as well as prediction of the response to pCRT. The results of DKI should be considered in treatment decision-making during the work-up of patients with rectal cancer.


Detailed voltammetric and EPR study of protonation reactions accompanying the one-electron reduction of Keggin-type polyoxometalates, [XV(V)M11O40]4- (X = P, As; M = Mo, W) in acetonitrile.
The one electron electrochemical reduction of vanadium(V) substituted-Keggin type polyoxometalate anions [XV(V)M(11)O(40)](4-) (XV(V)M(11)) where X = P, As; M = Mo or W has been investigated in CH(3)CN as a function of acid concentration. EPR studies confirm that the product is the highly basic and hence readily protonated V(IV), (XV(IV)M(11)), species. In the absence of acid or in the presence of a large concentration excess, the V(V/IV) redox couple gives one well defined chemically and electrochemically reversible process under conditions of cyclic voltammetry. In contrast, three well resolved V(V/IV) processes are observed in the presence of a moderate concentration of acid in the case of XV(V)Mo(11) but only two with XV(V)W(11). NMR and EPR spectra have been obtained as a function of acid concentration when the polyoxometalate anions are in the V(V) and V(IV) oxidation levels respectively. All voltammetric and spectroscopic data indicate that protonation reactions are coupled to the V(V) and V(IV) redox chemistry but that the reduced V(IV) state is much more basic than the V(V) one and that (XV(IV)Mo(11)) is more basic than (XV(IV)W(11)). Digital simulations of voltammograms for reduction of XV(V)Mo(11) and XV(V)W(11) have been undertaken in CH(3)CN as a function of acid concentration in order to define the thermodynamics and kinetics associated with the V(V/IV) process and the equilibrium constants that accompany the coupled acid-base chemistry. EPR spectra allow an estimation of the relative concentrations of protonated species present in frozen glasses derived from one-electron bulk electrolysis and also allow inferences to be drawn on their nature. This study provides a far more detailed analysis of the coupling of proton and electron transfer than previously available.


Diagnostic value of computed tomography scanning in differentiating malignant from benign solitary pulmonary nodules: a meta-analysis.
An early diagnosis of lung cancer is crucial for early treatment and management. The objective of this systematic review was to assess the overall diagnostic accuracy of chest computed tomography (CT) scanning in differentiating malignant from benign solitary pulmonary nodules (SPNs) with meta-analysis. The PubMed and China National Knowledge Infrastructure (CNKI) database were searched for eligible studies published up to March 2014. The sensitivity, specificity, and other measures of accuracy of chest CT scanning in the diagnosis of SPNs were pooled along with 95 % confidence intervals (CI). Summary receiver operating characteristic (ROC) curves were used to summarize overall test performance. Thirty-two studies met our inclusion criteria. The summary estimates for chest CT scanning in the diagnosis of SPNs in the meta-analysis were as follows: pooled sensitivity, 0.89 (95 % CI, 0.88 to 0.91); pooled specificity, 0.70 (95 % CI, 0.68 to 0.73); positive likelihood ratio, 2.88 (95 % CI, 2.46 to 3.37); negative likelihood ratio, 0.16 (95 % CI, 0.12 to 0.21); and diagnostic odds ratio, 23.83 (95 % CI, 16.18 to 35.11). The results indicate that CT scanning has relatively high sensitivity and moderate specificity for the diagnosis of SPNs. Given the low cost and growing prevalence of the technology, CT scanning should be recommended as the initial test for the evaluation of SPNs.


Spinal antinociception by dexmedetomidine, a highly selective alpha 2-adrenergic agonist.
The antinociceptive effects of dexmedetomidine, a highly selective new alpha 2-adrenoceptor agonist, were evaluated in rats after intrathecal, intraperitoneal and subcutaneous administration. Antinociception was tested using the tail-flick method. Both 3 and 6 micrograms of intrathecal dexmedetomidine produced maximal antinociception within 10 min. The effect lasted for up to 6 hr. The smaller dose of 1.5 micrograms produced a mean antinociception of 50% (of the maximum possible effect, MPE%) which lasted for about 2 hr. Subcutaneous atipamezole, a specific alpha 2-adrenergic antagonist completely abolished the antinociception produced by intrathecal dexmedetomidine. When given intraperitoneally, dexmedetomidine produced on average a 50% antinociceptive effect with the highest dose of 60 micrograms/kg. The lower doses of 10 and 30 micrograms/kg were ineffective. After subcutaneous administration a maximal effect was achieved with 120 micrograms/kg, a 70% effect, on average, with 60 micrograms/kg and a short lasting effect of 60% with 30 micrograms/kg. In conclusion, dexmedetomidine is a very potent antinociceptive agent when given intrathecally to rats.


Characterization of the potential energy surfaces of two small but challenging noncovalent dimers: (P2 )2 and (PCCP)2.
This work characterizes eight stationary points of the P2 dimer and six stationary points of the PCCP dimer, including a newly identified minimum on both potential energy surfaces. Full geometry optimizations and corresponding harmonic vibrational frequencies were computed with the second-order Møller-Plesset (MP2) electronic structure method and six different basis sets: aug-cc-pVXZ, aug-cc-pV(X+d)Z, and aug-cc-pCVXZ where X = T, Q. A new L-shaped structure with C2 symmetry is the only minimum for the P2 dimer at the MP2 level of theory with these basis sets. The previously reported parallel-slipped structure with C2 h symmetry and a newly identified cross configuration with D2 symmetry are the only minima for the PCCP dimer. Single point energies were also computed using the canonical MP2 and CCSD(T) methods as well as the explicitly correlated MP2-F12 and CCSD(T)-F12 methods and the aug-cc-pVXZ (X = D, T, Q, 5) basis sets. The energetics obtained with the explicitly correlated methods were very similar to the canonical results for the larger basis sets. Extrapolations were performed to estimate the complete basis set (CBS) limit MP2 and CCSD(T) binding energies. MP2 and MP2-F12 significantly overbind the P2 and PCCP dimers relative to the CCSD(T) and CCSD(T)-F12 binding energies by as much as 1.5 kcal mol(-1) for the former and 5.0 kcal mol(-1) for the latter at the CBS limit. The dominant attractive component of the interaction energy for each dimer configuration was dispersion according to several symmetry-adapted perturbation theory analyses.


Effects of estradiol on release and disposition of norepinephrine from nerve endings.
Studies were performed to determine whether estradiol (E2) has a direct action on the release and disposition of norepinephrine (NE) from adrenergic nerve endings in isolated superfused canine saphenous veins. [3H]NE and is labeled metabolites were separated by column chromatography with measurement by liquid scintillation spectrometry. An increase in the spontaneous overflow of total 3H, [3H]NE, and [3H]dihydroxyphenylglycol ([3H]DOPEG) that occurred with 1 and 10 microgram/ml E2 in the superfusing medium suggested that E2 either induced NE release or interfered with intraneuronal NE storage. During electrical stimulation (ES), release of [3H]NE and [3H]DOPEG exceeded controls with 1 and 10 microgram/ml E2 in the superfusate, yet total 3H was little changed. The evoked release of [3H]DOPEG showed less of an elevation over its spontaneous efflux in E2-treated veins than in nontreated veins, suggesting that E2 may block neuronal reuptake of released NE. The efflux of O-methylated deaminated metabolites during and after ES was decreased by E2 treatment, suggesting also an inhibition of extraneuronal uptake of NE.


Warming at the population level: Effects on age structure, density, and generation cycles.
The impact of climate change on strongly age-structured populations is poorly understood, despite the central role of temperature in determining developmental rates in ectotherms. Here we examine the effect of warming and its interactions with resource availability on the population dynamics of the pyralid moth Plodia interpunctella, populations of which normally show generation cycles, a consequence of strong and asymmetric age-related competition. Warming by 3°C above the standard culture temperature led to substantial changes in population density, age structure, and population dynamics. Adult populations were some 50% larger in warmed populations, probably because the reduced fecundity associated with warming leads to reduced larval competition, allowing more larvae to develop to adulthood. Warming also interacted with resource availability to alter population dynamics, with the generation cycles typical of this species breaking down in the 30° populations when standard laboratory diet was provided but not when a reduced nutrient poor diet was used. Warming by 6° led to either rapid extinction or the persistence of populations at low densities for the duration of the experiment. We conclude that even moderate warming can have considerable effects on population structure and dynamics, potentially leading to complete changes in dynamics in some cases. These results are particularly relevant given the large number of economically important species that exhibit generation cycling, in many cases arising from similar mechanisms to those operating in P. interpunctella.


[New diagnostic strategies in psychiatry by means of video-technique. I. The use of time-blind video analysis for the evaluation of antidepressant drug trials (author's transl)].
The use of audio-video methods allowed the decrease of subjective factors influencing ratings of psychopathological phenomina. TV-stored interviews, recorded at separate days, may be compared almost simultaneously. In this study a rating of severity of depression was made of patients under antidepressive medication on days 0, 10, 20. (Mianserin was compared with Amitriptyline in a double-blind-trial). This method rendered it possible to detect significant differences between therapeutic profiles over time. The same result was reached independently by both investigators. The judgement of depression was based on a global impression and not on the evaluation of isolated psychopathological symptoms. No absolute measurement of depression was possible. The described method of a time-blind-analysis of TV-stored interviews constitutes a considerable progress in the assessment of a drug's profile in time, particularly if rating-scales are used in the same manner.


Minor head trauma in soccer and serum levels of S100B.
To compare the serum levels of S100B after head trauma with the effect of heading, high-intensity exercise, and playing in a league match. Heading and head trauma in soccer have been suspected to cause brain impairment. The protein S100B is a marker of acute neuronal tissue damage. Baseline S100B was measured in 535 Norwegian professional soccer players. Two hundred twenty-eight head impacts were registered from 352 league matches. Three teams (n = 48) performed a high-intensity exercise session without heading and a low-intensity session with heading exercises. A blood sample was drawn from each participant within 1 hour (B1) after the session, and another sample (B12) was drawn after a match or training session. The players were assigned to four groups: Head Impact (n = 65), Match Control (match participants without head impact, n = 49), High-intensity Exercise (n = 35), and Heading (n = 36). Serum S100B increased from baseline to B1 for all groups. The increase for the match groups (Head Impact and Match Control) was significantly higher than for both training groups. However, no significant differences between the Head Impact and Match Control groups or between the two training groups were found. A total of 39 players (33.9%) had elevated B1 values (>/=0.12 ng/ml) after a match, but these findings were equally distributed between the Match Control and Head Impact groups. Both soccer training and soccer matches cause a transient increase in S100B. There is a possible additive effect of activity with high intensity and heading, but minor head impacts do not seem to cause an additional increase.


Association of Unexpected Newborn Deaths With Changes in Obstetric and Neonatal Process of Care.
The death of a healthy term infant may signal patient safety and quality issues. Various initiatives aim to encourage clinicians to learn from these events, but little evidence exists regarding how exposure to an unexpected newborn death may alter clinician practice. To examine the association between an unexpected newborn death and changes in obstetric and newborn procedures that may be used in response to potential fetal distress or newborn complications. This cross-sectional study used difference-in-differences analysis of 2011 to 2017 US vital statistics data from 477 US counties experiencing an unexpected newborn death during the study period. All in-hospital live births in the 477 counties during the study period were included. Data were analyzed from September 2019 to September 2020. The death of an infant aged 0 to 7 days following an unremarkable pregnancy owing to causes other than birth defects, accidents/assaults, or sudden infant death syndrome. Primary outcomes included binary variables capturing intervention in labor/delivery (induction, augmentation, cesarean delivery, forceps/vacuum) and procedures to avert and mitigate newborn complications (assisted ventilation, surfactant replacement therapy, antibiotics for suspected sepsis, neonatal intensive care unit admission). The main sample included 5.72 million births (2.54 million during preexposure time). Mean (SD) maternal age was 27.3 (5.8) years; 67% of mothers were White, and 12% were Black. Associations varied across the 4 estimated models. Following an unexpected newborn death, there was no significant increase in the probability of cesarean delivery in the full sample model (0.28 percentage points [pp]; 95% CI, -0.01 to 0.57 pp), but a significant increase in the other 3 models, with values ranging from 0.55 pp (95% CI, 0.21 to 0.88 pp) in the full sample model with matching to 0.66 pp (95% CI, 0.13 to 1.19 pp) in the 1-hospital county subsample with matching. There was a significant increase in the probability of newborn assisted ventilation in the full sample model with matching (0.46 pp; 95% CI, 0.08 to 0.83 pp), but no significant increase in the other 3 models, with estimates ranging from 0.33 pp (95% CI, -0.04 to 0.71 pp) to 0.69 pp (95% CI, -0.02 to 1.40 pp). An unexpected newborn death was not associated with a significant increase in antibiotic use in the full sample models (without matching: 0.19 pp; 95% CI, -0.00 to 0.39 pp; with matching: 0.22 pp; 95% CI: -0.02 to 0.46 pp), but was associated with a significant increase in both of the 1-hospital county subsample models (without matching: 0.38 pp; 95% CI, 0.02 to 0.73 pp; with matching: 0.39 pp; 95% CI, 0.01 to 0.77 pp). In some study models, an unexpected newborn death was associated with statistically significant increases in subsequent use of procedures to avert and mitigate fetal distress and newborn complications, which could reflect increases in identifying and proactively addressing serious potential complications or increased clinician caution applied across all cases. Future research should address whether these changes affect patient outcomes.


Cartilaginous differentiation in the joint capsule.
The proliferation and differentiation of cells are greatly influenced by their environment. Many growth factors and cytokines are reported to be environmental factors that affect the proliferation and differentiation of cells. Mechanical stress is also considered to influence these physiological reactions. The joint capsule, which is a part of the joint tissue, plays a very important role in the stability of the joint and in maintaining the intracapsular phenomenon. In patients with dislocated hip arthropathy, this capsule is involved in the weightbearing function by forming a sliding surface between the capsule and the femoral head articular cartilage. The surface of the tissue macroscopically shows cartilaginous change, which indicates cartilaginous differentiation caused by mechanical stress. We examined the cartilage-specific proteoglycan component, which is composed of cartilaginou matrix at the differentiation site. We investigated proteoglycan production, molecular size, and the gene expression of cartilaginous substrate. At the inner layer of the weightbearing area of the joint capsule, proteoglycan production was significantly higher than that of other noncartilaginous tissue. We also identified the gene expression of cartilaginous proteoglycan using the reverse transcription polymerase chain reaction (RT-PCR) method.


Endotoxin challenge causes a proinflammatory state in obstructive jaundice.
The aim of this study was to investigate the relationship between the obstructive jaundice-induced cellular immune suppression and endotoxin challenge with respect to the levels of tumor necrosis factor (TNF), interleukin-10 (IL-10), and interleukin-2 (IL-2). Rats underwent either bile duct ligation or sham operation. At 21 days, all rats were challenged either with lipopolysaccharide (LPS) or saline. In the sham-operated group LPS injection significantly increased TNF levels at 90 min. The common bile duct ligated group showed a significant increase in TNF levels compared with all other groups, including the sham-operated, LPS-injected group, at 90 min. At 180 min following LPS challenge, TNF levels decreased, and there was no difference between any of the LPS-challenged groups at 180 min and any of the saline groups at either 90 or 180 min. In the sham-operated group, LPS injection significantly increased IL-10 levels at both 90 and 180 min. In the bile duct ligated group, LPS injection significantly increased IL-10 levels compared with saline injection at both 90 and 180 min. On the other hand, bile duct ligated animals had significantly less increase in IL-10 levels following LPS challenge at 90 min but not at 180 min. In common bile duct ligated rats, LPS challenge induced a significantly greater increase in IL-2 levels compared with all other groups. In conclusion, in the presence of obstructive jaundice, endotoxemia primes a more vigorous inflammatory response despite cellular immune depression.


Characterisation of a second gain of function EDAR variant, encoding EDAR380R, in East Asia.
Ectodysplasin A1 receptor (EDAR) is a TNF receptor family member with roles in the development and growth of hair, teeth and glands. A derived allele of EDAR, single-nucleotide variant rs3827760, encodes EDAR:p.(Val370Ala), a receptor with more potent signalling effects than the ancestral EDAR370Val. This allele of rs3827760 is at very high frequency in modern East Asian and Native American populations as a result of ancient positive selection and has been associated with straighter, thicker hair fibres, alteration of tooth and ear shape, reduced chin protrusion and increased fingertip sweat gland density. Here we report the characterisation of another SNV in EDAR, rs146567337, encoding EDAR:p.(Ser380Arg). The derived allele of this SNV is at its highest global frequency, of up to 5%, in populations of southern China, Vietnam, the Philippines, Malaysia and Indonesia. Using haplotype analyses, we find that the rs3827760 and rs146567337 SNVs arose on distinct haplotypes and that rs146567337 does not show the same signs of positive selection as rs3827760. From functional studies in cultured cells, we find that EDAR:p.(Ser380Arg) displays increased EDAR signalling output, at a similar level to that of EDAR:p.(Val370Ala). The existence of a second SNV with partly overlapping geographic distribution, the same in vitro functional effect and similar evolutionary age as the derived allele of rs3827760, but of independent origin and not exhibiting the same signs of strong selection, suggests a northern focus of positive selection on EDAR function in East Asia.


End-stage heart failure: Two surgical approaches with different rehabilitative outcomes.
A rising number of patients are surgically treated for heart failure at the more advanced stage, thanks to the increasing use of left ventricular assist device (LVAD) as a reliable alternative to heart transplantation (HTx). However, it is still unknown whether differences exist between the two surgical approaches in the efficacy of rehabilitation programmes. Therefore, aim of this study was to evaluate whether functional capacity and rehabilitative outcomes differ between HTx and implantation of LVAD. We enrolled 51 patients with HTx and 46 with LVAD upon admission to our rehabilitation-unit. We evaluated six-minute walking test (6MWT), resting oxygen saturation (SaO2) and nutritional assessment before and after a standardised cardiovascular rehabilitation programme. HTx and LVAD groups differed in age, anthropometric variables, gender distribution. Upon enrolment, 6MWT distance was similar in the two groups, whereas malnutrition was less frequent and the waist circumference/height ratio (WHtR) was greater in LVAD patients. SaO2 was greater in HTx patients. Rehabilitation improved SaO2, 6MWT distance and nutritional status. The difference in malnutrition disappeared, but WHtR remained higher in the LVAD and SaO2 higher in the HTx patients; the 6MWT distance improved more in the HTx patients. Multivariate linear regression analysis confirmed that the type of intervention was independent predictor of 6MWT distance after rehabilitation. HTx patients improve more rapidly and perform better after rehabilitation, suggesting the need for more tailored rehabilitation training for LVAD patients.


Clozapine treatment for neuroleptic-induced tardive dyskinesia, parkinsonism, and chronic akathisia in schizophrenic patients.
Previous studies on the use of clozapine in neuroleptic-resistant chronic schizophrenic patients have demonstrated positive effects on tardive dyskinesia but were less conclusive about chronic akathisia and parkinsonism. The aim of the present study was to investigate the short-term (18 weeks) efficacy of clozapine in neuroleptic-resistant chronic schizophrenic patients with coexisting tardive dyskinesia, chronic akathisia, and parkinsonism. Twenty chronic, neuroleptic-resistant schizophrenic patients with coexisting tardive dyskinesia, parkinsonism, and chronic akathisia were treated with clozapine. Assessment of tardive dyskinesia, parkinsonism, and chronic akathisia was made once weekly for 18 weeks with the Abnormal Involuntary Movement Scale (AIMS), Simpson-Angus Rating Scale for Extrapyramidal Side Effects, and Barnes Rating Scale for Drug-Induced Akathisia (BAS). At the end of 18 weeks of clozapine treatment, improvement rates were 74% for tardive dyskinesia, 69% for parkinsonism, and 78% for chronic akathisia. A statistically significant reduction in the scores on the AIMS and Simpson-Angus Scale was achieved at Week 5 and on the BAS at Week 6 (p < .0001). Relatively low doses of clozapine are effective for the treatment of neuroleptic-induced extrapyramidal syndromes in neuroleptic-resistant chronic schizophrenic patients. The relief of tardive dyskinesia, parkinsonism, and chronic akathisia in this group of patients occurs more rapidly than the reduction in psychotic symptoms. Disturbing, long-term extrapyramidal syndromes in chronic schizophrenic patients should be considered an indication for clozapine treatment.


Maximal eccentric and concentric strength discrepancies between young men and women for dynamic resistance exercise.
Although research has demonstrated that isokinetic eccentric (ECC) strength is 20-60% greater than isokinetic concentric (CON) strength, few data exist comparing these strength differences in standard dynamic resistance exercises. The purpose of the study was to determine the difference in maximal dynamic ECC and CON strength for 6 different resistance exercises in young men and women. Ten healthy young men (mean +/- SE, 25.30 +/- 1.34 years), and 10 healthy young women (mean +/- SE, 23.40 +/- 1.37 years) who were regular exercisers with resistance training experience participated in the study. Two sessions were performed to determine CON and ECC 1 repetitions maximum for latissimus pull-down (LTP), leg press (LP), bench press (BP), leg extension (LE), seated military press (MP), and leg curl (LC) exercises. Maximal ECC and maximal CON strength were determined on weight stack machines modified to isolate ECC and CON contractions using steel bars and pulleys such that only 1 type of contraction was performed. Within 2 weeks, participants returned and completed a retest trial in a counterbalanced fashioned. Test-retest reliability was excellent (r = 0.99) for all resistance exercise trials. Men demonstrated 20-60% greater ECC than CON strength (LTP = 32%, LP = 44%, BP = 40%, LE = 35%, MP = 49%, LC = 27%). Women's strength exceeded the proposed parameters for greater ECC strength in 4 exercises, p < 0.05 (LP = 66%, BP = 146%, MP = 161%, LC = 82%). The ECC/CON assessment could help coaches capitalize on muscle strength differences in young men and women during training to aid in program design and injury prevention and to enhance strength development.


Tumor classification ranking from microarray data.
Gene expression profiles based on microarray data are recognized as potential diagnostic indices of cancer. Molecular tumor classifications resulted from these data and learning algorithms have advanced our understanding of genetic changes associated with cancer etiology and development. However, classifications are not always perfect and in such cases the classification rankings (likelihoods of correct class predictions) can be useful for directing further research (e.g., by deriving inferences about predictive indicators or prioritizing future experiments). Classification ranking is a challenging problem, particularly for microarray data, where there is a huge number of possible regulated genes with no known rating function. This study investigates the possibility of making tumor classification more informative by using a method for classification ranking that requires no additional ranking analysis and maintains relatively good classification accuracy. Microarray data of 11 different types and subtypes of cancer were analyzed using MDR (Multi-Dimensional Ranker), a recently developed boosting-based ranking algorithm. The number of predictor genes in all of the resulting classification models was at most nine, a huge reduction from the more than 12 thousands genes in the majority of the expression samples. Compared to several other learning algorithms, MDR gives the greatest AUC (area under the ROC curve) for the classifications of prostate cancer, acute lymphoblastic leukemia (ALL) and four ALL subtypes: BCR-ABL, E2A-PBX1, MALL and TALL. SVM (Support Vector Machine) gives the highest AUC for the classifications of lung, lymphoma, and breast cancers, and two ALL subtypes: Hyperdiploid > 50 and TEL-AML1. MDR gives highly competitive results, producing the highest average AUC, 91.01%, and an average overall accuracy of 90.01% for cancer expression analysis. Using the classification rankings from MDR is a simple technique for obtaining effective and informative tumor classifications from cancer gene expression data. Further interpretation of the results obtained from MDR is required. MDR can also be used directly as a simple feature selection mechanism to identify genes relevant to tumor classification. MDR may be applicable to many other classification problems for microarray data.


Commotio cordis in non-sports-related injury: A scoping review.
Commotio cordis is a rare but life-threatening condition characterized by sudden cardiac arrest resulting from a blunt chest impact. While commotio cordis has traditionally been associated with sports-related activities, a significant proportion of cases occur in non-sport-related settings, such as assaults, motor vehicle accidents (MVAs), and daily activities. This critical review examines the epidemiology, clinical characteristics, and outcomes of non-sports-related commotio cordis cases, highlighting the need for increased awareness and improved management in these contexts. The review analyzes existing literature, drawing attention to the demographics of non-sports-related cases, which predominantly affect adolescents and young adults, with males being the primary demographic. In contrast to sport-related cases, non-sports-related commotio cordis cases exhibit a wider age range and a higher proportion of female subjects. Mortality rates are significantly higher in non-sports-related commotio cordis cases, largely due to lower rates of cardiopulmonary resuscitation (CPR), limited access to automated external defibrillators (AEDs), and delayed initiation of resuscitative efforts compared to sport-related incidents. This underscores the critical importance of increasing awareness and preparedness in non-sport-related settings. To mitigate the risks associated with non-sports-related commotio cordis, efforts should focus on early recognition of the condition, timely administration of CPR, and the widespread availability and accessibility of AEDs in various environments. Enhanced awareness and education can potentially lead to a reduction in mortality and improved outcomes for individuals affected by commotio cordis outside of sports-related activities. In conclusion, commotio cordis is not exclusive to sports and presents a significant health risk in non-sport-related scenarios. This review emphasizes the urgent need for increased awareness, preparedness, and resuscitation measures in non-sports contexts to address the higher mortality associated with these cases.


The phyllolepid placoderm Cowralepis mclachlani: insights into the evolution of feeding mechanisms in jawed vertebrates.
Remarkably preserved specimens of Cowralepis mclachlani Ritchie, 2005 (Proc Linn Soc NSW 126:215-259) (Phyllolepida, Placodermi) represent a unique ontogenetic sequence adding to our understanding of anatomy, function, and phylogeny among basal jawed vertebrates (gnathostomes). A systematic review demonstrates that the Phyllolepida are a subgroup of the Arthrodira. Consideration of visceral and neurocranial characters supports the hypothesis that placoderms are the sister group to remaining gnathostomes. Placoderms possess, as adult plesiomorphic features, a number of characters that are only seen in the development of extant gnathostomes-a peramorphic shift relative to placoderms. Developmental evidence in vertebrates leads to a revised polarity of character transitions. These include 1) hyomandibula-neurocranium and ventral parachordal-palatoquadrate articulations (vertebrate synapomorphies); 2) jointed pharynx, paired basibranchials, anterior ethmoidal-palatoquadrate articulation, short trabeculae cranii, and anterior and posterior neurocranial fissures (gnathostome synapomorphies); and 3) fused basibranchials, dorsal palatoquadrate-neurocranium articulation, loss of the anterior neurocranial fissure, elongated trabeculae cranii, and transfer of the ventral parachordal-palatoquadrate articulation to the trabeculae (crown group gnathostomes). The level of preservation in C. mclachlani provides the basis for a reinterpretation of phyllolepid anatomy and function. Cowralepis mclachlani possesses paired basibranchials allowing the reinterpretation of the visceral skeleton in other placoderms. Mandible depression in C. mclachlani follows an osteichthyan pattern and the ventral visceral skeleton acts as a functional unit. Evidence for hypobranchial musculature demonstrates the neural crest origin of the basibranchials and that Cowralepis was a suction feeder. Finally, the position of the visceral skeleton relative to the neurocranium in placoderms parallels the condition in selachians and osteichthyans, but differs in the elongation of the occiput. The cucullaris fossa of placoderms (interpreted as a site of muscle attachment) is shown to represent, in part, the parabranchial chamber.


Sigma(B)-dependent expression patterns of compatible solute transporter genes opuCA and lmo1421 and the conjugated bile salt hydrolase gene bsh in Listeria monocytogenes.
Listeria monocytogenes is a food-borne pathogen that can persist and grow under a wide variety of environmental conditions including low pH and high osmolarity. The alternative sigma factor sigma(B) contributes to L. monocytogenes survival under extreme conditions. The purpose of this study was to identify and confirm specific sigma(B)-dependent genes in L. monocytogenes and to characterize their expression patterns under various stress conditions. opuCA, lmo1421 and bsh were identified as putative sigma(B)-dependent genes based on the presence of a predicted sigma(B)-dependent promoter sequence upstream of each gene. opuCA and lmo1421 encode known and putative compatible solute transporter proteins, respectively, and bsh encodes a conjugated bile salt hydrolase (BSH). Reporter fusions and semi-quantitative RT-PCR techniques were used to confirm sigma(B)-dependent regulation of these stress-response genes and to determine their expression patterns in response to environmental stresses. RT-PCR demonstrated that opuCA, lmo1421 and bsh transcript levels are reduced in stationary-phase L. monocytogenes deltasigB cells relative to levels present in wild-type cells. Furthermore, BSH activity is abolished in a L. monocytogenes deltasigB strain. RT-PCR confirmed growth-phase-dependent expression of opuCA, with highest levels of expression in stationary-phase cells. The L. monocytogenes wild-type strain exhibited two- and threefold induction of opuCA expression and seven- and fivefold induction of lmo1421 expression following 10 and 15 min exposure to 0.5 M KCl, respectively, as determined by RT-PCR, suggesting rapid induction of sigma(B) activity in exponential-phase L. monocytogenes upon exposure to salt stress. Single-copy chromosomal opuCA-gus reporter fusions also showed significant induction of opuCA expression following exposure of exponential-phase cells to increased salt concentrations (0.5 M NaCl or 0.5 M KCl). In conjunction with recent findings that indicate a role for opuCA and bsh in L. monocytogenes virulence, the data presented here provide further evidence of specific sigma(B)-mediated contributions to both environmental stress resistance and intra-host survival in L. monocytogenes.


[Molecular cloning,nucleotides sequence and transient expression of S and M genome segment of hantavirus strain 84Fli].
To sequence, analyze and express the nucleotide sequences of S and M segments of hantavirus strain 84Fli. S and M segments of hantavirus 84Fli strain were amplified by RT-PCR, the PCR products were cloned into plasmid pCR2.1-TOPOr. Three clones of each segment which have been sequenced were randomly selected. The coding region of S and M segments were amplified by PCR, and cloned into expressing vector pcDNA3.0. Transient expression of nucleocapsid protein, G1 and G2 glycoproteins in COS7 cells were performed by Lipofectin transfection. The expression of NP, G1 and G2 have been conformed by using immunofluorescence, Western blot and immuno-precipitation methods. The full length S and M segments cDNA have been amplified and cloned. The S and M segments of hantavirus strain 84Fli contained 1 688 and 3 616 nucleotides respectively. Deduced amino acid sequences of NP and glycoproteins revealed high homologue to other Hantaan viruses. NP, G1 and G2 proteins of 84Fli can be transiently expressed in COS7 cells.


Low density lipoprotein receptor-related protein-2 expression in efferent duct and epididymal epithelia: evidence in rats for its in vivo role in endocytosis of apolipoprotein J/clusterin.
Apolipoprotein J/clusterin/sulfated glycoprotein-2 (apo J) disassociates from spermatozoa and is endocytosed by epithelial cells lining the efferent ducts and epididymis. The low density lipoprotein receptor-related protein-2/megalin (LRP-2) has been shown to bind to apo J and mediates its endocytosis and lysosomal degradation in cultured cells. In this study, immunocytological techniques were used to localize LRP-2 in rat efferent ducts and epididymis and to determine whether its expression correlated with those epithelial cells involved in apo J endocytosis. Pronounced LRP-2 immunochemical staining was observed on the apical surfaces of epithelial cells lining the efferent ducts and in the intermediate zone, proximal caput, and corpus and cauda regions of the epididymis. Single immunogold labeling at the electron microscopic level showed LRP-2 to be present within coated pits, endocytic vesicles, and early endosomes of the nonciliated cells of the efferent ducts and the principal cells of the epididymis. In efferent ducts, double immunogold labeling showed both LRP-2 and apo J to be present in endocytic compartments including coated pits, endocytic vesicles, and early endosomes of nonciliated cells. However, while apo J was detected in late endosomes and lysosomes of nonciliated cells, LRP-2 was not. Apical tubules, possibly emerging from late endosomes, contained labeling for LRP-2 but not for apo J. Ciliated cells lying adjacent to nonciliated cells displayed no labeling for either LRP-2 or apo J. These results are consistent with the possibility that LRP-2 serves as an endocytic receptor for apo J in vivo and that after endocytosis the LRP-2 is recycled back to the cell surface while apo J is delivered to the lysosomes for degradation. To provide additional evidence implicating LRP-2 in apo J endocytosis, a receptor-associated protein (RAP), an antagonist of apo J binding to LRP-2, was injected into the efferent duct lumen. Subsequent immunocytological analysis of the efferent duct showed that the RAP treatment abolished the endocytosis of apo J by the nonciliated cells. Taken together, these data indicate that LRP-2 is a likely mediator of apo J endocytosis by the nonciliated efferent duct cells.


A prospective phase II study of pre-operative chemotherapy then short-course radiotherapy for high risk rectal cancer: COPERNICUS.
Neoadjuvant chemotherapy (NAC) allows earlier treatment of rectal cancer micro-metastases but is not standard of care. There are currently no biomarkers predicting long-term progression-free survival (PFS) benefit from NAC. In this single arm phase II trial, patients with non-metastatic magnetic resonance imaging (MRI)-defined operable rectal adenocarcinoma at high risk of post-operative metastatic recurrence, received 8 weeks of oxaliplatin/fluorouracil NAC then short-course preoperative radiotherapy (SCPRT) before immediate surgery. Sixteen weeks of post-operative adjuvant chemotherapy (AC) was planned. A pelvic MRI was performed at week 9 immediately post-NAC, before SCPRT. The primary end point was feasibility assessed by completion of protocol treatment up to and including surgery. Secondary endpoints included compliance, toxicity, downstaging efficacy, and PFS. In total 60 patients were recruited May 2012-June 2014. In total 57 patients completed protocol treatment, meeting the primary endpoint. Compliance with NAC was much better than AC: Comparing NAC vs. AC, the median percentage dose intensity for fluoropyrimidine was 100% vs. 63% and for oxaliplatin 100% vs. 45%. Treatment-related toxicity was acceptable with no treatment-related deaths. Post-NAC MRI showed 44 tumours (73%) were T-downstaged and 22 (37%) had excellent MRI tumour regression grade (mrTRG 1-2). Median follow-up was 27 months with 2-year PFS of 86.2% (10 events). On exploratory analysis, post-NAC mrTRG predicted PFS with no event among those with excellent regression. The regimen was well tolerated with effective downstaging and encouraging PFS. mrTRG response to NAC may be a new prognostic factor for long-term PFS, but needs validation in larger studies.


Effect of vitamin D replacement on hip structural geometry in adolescents: a randomized controlled trial.
We have shown in a randomized controlled trial that vitamin D increases bone mass, lean mass and bone area in adolescent girls, but not boys. These increments may translate into improvements in bone geometry and therefore bone strength. This study investigated the impact of vitamin D on hip geometric dimensions from DXA-derived hip structural analyses in adolescents who participated in the trial. 167 girls (mean age 13.1 years) and 171 boys (mean age 12.7 years) were randomly assigned to receive weekly placebo oil or vitamin D3, at doses of 1400 IU or 14,000 IU, in a double blind placebo-controlled 1-year trial. DXA images were obtained at baseline and one year, and hip images were analyzed using the hip structural analysis (HSA) software to derive parameters of bone geometry. These include outer diameter (OD), cross sectional area (CSA), section modulus (Z), and buckling ratio (BR) at the narrow neck (NN), intertrochanteric (IT), and shaft (S) regions. Analysis of Covariance (ANCOVA) was used to examine group differences for changes of bone structural parameters. In the overall group of girls, vitamin D supplementation increased aBMD (7.9% and 6.8% in low and high doses, versus 4.2% in placebo) and reduced the BR of NN (6.1% and 2.4% in low and high doses, versus 1.9% in placebo). It also improved aBMD (7.9% and 5.2% versus 3.6%) and CSA (7.5% and 5.1% versus 4.1%) of the IT and OD of the S (2.4% and 2.5% versus 0.8% respectively). Significant changes in the OD and BR of the NN, in the overall group of girls remained, after adjusting for lean mass, and were unaffected with further adjustments for lifestyle, pubertal status, and height measures. Conversely, boys did not exhibit any significant changes in any parameters of interest. A dose effect was not detected and subgroup analyses revealed no beneficial effect of vitamin D by pubertal stage. Vitamin D supplementation improved bone mass and several DXA-derived structural bone parameters, in adolescent girls, but not boys. This occurred at a critical site, the femoral neck, and if maintained through adulthood could improve bone strength and lower the risk of hip fractures.


Hydrocortisone requirement for the induction of glutamine synthetase in chick-embryo retinas.
Hydrocortisone has been found to induce glutamine synthetase activity in chick-embryo retinas in culture. Evidence is presented to show that the hydrocortisone is definitely required for transcription; its requirement for translation has not been ruled out. The possible identity of hydrocortisone with an active component of calf-serum diffusate reported earlier is discussed. The data also indicate that the glutamine synthetase messenger RNA is stable for at least several hours.


Pressure ulcer prevention algorithm content validation: a mixed-methods, quantitative study.
Translating pressure ulcer prevention (PUP) evidence-based recommendations into practice remains challenging for a variety of reasons, including the perceived quality, validity, and usability of the research or the guideline itself. Following the development and face validation testing of an evidence-based PUP algorithm, additional stakeholder input and testing were needed. Using convenience sampling methods, wound care experts attending a national wound care conference and a regional wound ostomy continence nursing (WOCN) conference and/or graduates of a WOCN program were invited to participate in an Internal Review Board-approved, mixed-methods quantitative survey with qualitative components to examine algorithm content validity. After participants provided written informed consent, demographic variables were collected and participants were asked to comment on and rate the relevance and appropriateness of each of the 26 algorithm decision points/steps using standard content validation study procedures. All responses were anonymous. Descriptive summary statistics, mean relevance/appropriateness scores, and the content validity index (CVI) were calculated. Qualitative comments were transcribed and thematically analyzed. Of the 553 wound care experts invited, 79 (average age 52.9 years, SD 10.1; range 23-73) consented to participate and completed the study (a response rate of 14%). Most (67, 85%) were female, registered (49, 62%) or advanced practice (12, 15%) nurses, and had > 10 years of health care experience (88, 92%). Other health disciplines included medical doctors, physical therapists, nurse practitioners, and certified nurse specialists. Almost all had received formal wound care education (75, 95%). On a Likert-type scale of 1 (not relevant/appropriate) to 4 (very relevant and appropriate), the average score for the entire algorithm/all decision points (N = 1,912) was 3.72 with an overall CVI of 0.94 (out of 1). The only decision point/step recommendation with a CVI of ≤ 0.70 was the recommendation to provide medical-grade sheepskin for patients at high risk for friction/shear. Many positive and substantive suggestions for minor modifications including color, flow, and algorithm orientation were received. The high overall and individual item rating scores and CVI further support the validity and appropriateness of the PUP algorithm with the addition of the minor modifications. The generic recommendations facilitate individualization, and future research should focus on construct validation testing.


Myocardial contrast echocardiography in subarachnoid hemorrhage-induced cardiac dysfunction: case report.
Cardiac dysfunction is a well-known complication of aneurysmal subarachnoid hemorrhage (SAH) that is generally regarded as secondary to catecholamine excess rather than overt ischemia. Myocardial contrast echocardiography (MCE) is a novel method of evaluating cardiac function and perfusion. We report the use of MCE in a patient with SAH and correlate the results to coronary angiography. Bedside MCE using Definity contrast agent (Bristol-Myers Squibb/Sanofi Pharmaceuticals, New York, NY) was performed at the onset of SAH and at the 1-week and 4-month follow-up evaluations. A 64-year-old woman presented with aneurysmal SAH. She developed transient ST elevation on lateral electrocardiographic leads and elevated cardiac enzymes with creatine-kinase MB isoenzyme of 44.3 ng/ml and troponin of 0.62 ng/ml. An emergent coronary angiogram performed at the outside facility revealed normal coronary anatomy, ejection fraction of 30%, and midventricular akinesis. On transfer to our facility, MCE demonstrated an ejection fraction of 45% with normal coronary perfusion in the akinetic midventricular segments and normally contracting basal and apical segments. At the 4-month follow-up examination, her ejection fraction normalized to 67% and regional wall motion had improved. To our knowledge, our case represents the first reported use of MCE in a patient with SAH. MCE demonstrating normal myocardium perfusion in the setting of normal coronary arteries on coronary angiogram and midventricular akinetic segments is compatible with nonischemic injury, which further supports the "catecholamine hypothesis" of neurogenic cardiac stunning. MCE may be a feasible noninvasive method to evaluate myocardial perfusion in the SAH population.


Fifty Years of Research in ARDS. Long-Term Follow-up after Acute Respiratory Distress Syndrome. Insights for Managing Medical Complexity after Critical Illness.
Critical illness is not a discrete disease state or syndrome. It is the culmination of a multiplicity of heterogeneous disease states and their varied health trajectories leading to extreme illness that requires advanced life support in a distinct geographic location in the hospital. It is a marker of newly acquired or worsened medical complexity and multimorbidities. Fifty years ago, distinguished critical care colleagues identified a syndrome of severe lung injury that united a group of patients with disparate admitting diagnoses. Acute respiratory distress syndrome continues to represent an important, incremental insult and risk modifier of acute and longer-term outcome, but it does not solely define our patients or their outcomes in isolation. Over the next 50 years, our research and clinical agenda needs to sharpen our lens on the fundamental importance of our patients' pre-critical illness health status, their intrinsic susceptibilities to tissue injury, and their innate and varied resiliencies. We need to take responsibility for the contribution that we make to morbidity through our practice in the intensive care unit each day. Engagement in frank and transparent communication with our patients and their caregivers about the very real and morbid consequences of being this sick is essential. We must enforce explicit consent about the morbidity of innovative, experimental, or high-risk medical and surgical procedures and ensure that our ongoing level of treatment aligns with patients' and caregivers' goals and values. Interprofessional and multidisciplinary collaboration is crucial to modify existing complex care pathways for our patients and their families to foster optimal rehabilitation and reintegration into the workplace and community.


Predictive factors for immunological and virological endpoints in Thai patients receiving combination antiretroviral treatment.
Routine CD4 count and HIV viral load monitoring is a financial barrier in developing countries. We assessed factors associated with CD4 counts < or =200 cells/microL and detectable viral load in Thai HIV-infected patients receiving antiretroviral therapy (ART) at the HIV Netherlands Australia Thailand Research Collaboration and the Thai Red Cross AIDS Research Centre (HIV-NAT). Univariate and multivariate Cox proportional hazards models for multiple treatment failures were used to determine factors related to CD4 counts < or =200 cells/microL and detectable viral load. Multivariate Cox proportional hazards models for CD4 counts < or =200 cells/microL were developed with and without viral load in order to build models applicable to contexts in which viral load is not available. Four hundred and seventeen patients were included in the study. Fifty-four per cent were male, and the median CD4 count and log(10) viral load at baseline were 283 cells/microL and 4.3 log(10) HIV-1 RNA copies/mL, respectively. Independent factors related to CD4 count < or =200 cells/microL were CD4 count at baseline [hazards ratio (HR) 0.20/100 cells/microL; 95% confidence interval (CI) 0.17-0.23] and changes in CD4 count (HR 0.22/100 cells/microL; 95% CI 0.17-0.28). Factors in multivariate models (in which viral load was considered for inclusion) were CD4 count at baseline (HR 0.21/100 cells/microL; 95% CI 0.18-0.24), changes in CD4 count (HR 0.25/100 cells/microL; 95% CI 0.19-0.32) and detectable viral load (HR 1.94; 95% CI 1.20-3.13). Predictive factors (independent of viral load) were triple ART or highly active antiretroviral therapy (HAART) (HR 0.28; 95% CI 0.22-0.36) and detectable viral load at baseline (HR 2.96; 95% CI 2.24-3.91). Conclusions CD4 count at baseline and changes in CD4 count were important in predicting CD4 counts < or =200 cells/microL. Triple ART and detectable viral load at baseline were important in predicting detectable viral load.


Identification of Neutrophil Extracellular Traps in Paraffin-Embedded Feline Arterial Thrombi using Immunofluorescence Microscopy.
Neutrophil extracellular traps (NETs), composed of cell-free DNA (cfDNA) and proteins like histones and neutrophil elastase (NE), are released by neutrophils in response to systemic inflammation or pathogens. Although NETs have previously been shown to augment clot formation and inhibit fibrinolysis in humans and dogs, the role of NETs in cats with cardiogenic arterial thromboembolism (CATE), a life-threatening complication secondary to hypertrophic cardiomyopathy, is unknown. A standardized method to identify and quantify NETs in cardiogenic arterial thrombi in cats will advance our understanding of their pathological role in CATE. Here, we describe a technique to identify NETs in formaldehyde-fixed and paraffin-embedded thrombi within the aortic bifurcation, extracted during necropsy. Following deparaffinization with xylene, aortic sections underwent indirect heat-induced antigen retrieval. Sections were then blocked, permeabilized, and ex vivo NETs were identified by colocalization of cell-free DNA (cfDNA), citrullinated histone H3 (citH3), and neutrophil elastase (NE) using immunofluorescence microscopy. To optimize the immunodetection of NETs in thrombi, autofluorescence of tissue elements was limited by using an autofluorescence quenching process prior to microscopy. This technique could be a useful tool to study NETs and thrombosis in other species and offers new insights into the pathophysiology of this complex condition.


Asymptomatic throat carriage rate and antimicrobial resistance pattern of Streptococcus pyogenes in Nepalese school children.
Streptococcus pyogenes or Group A streptococcus (GAS) causes several suppurative and non suppurative infections. In addition to pharyngitis and skin infections, GAS are also the causative agent of post-streptococcal infection syndromes such as acute rheumatic fever (ARF) and post-streptococcal glumerulonephritis (PSG). GAS frequently colonises in the throat of an asymptomatic person. Pharyngeal carriage rates of GAS among healthy school children vary with geographical location and seasons. We carried out this preliminary study to determine the throat carriage rate and antimicrobial resistance trend of Streptococcus pyogenes or Group A streptococcus (GAS) among the Nepalese school children. Four schools situated at different locations of Kathmandu valley were included in the study. Throat swabs from 350 students of age group 5-15 years were collected, immediately transported to the laboratory and were processed for S. pyogenes following standard microbiological procedures. Antimicrobial susceptibility testing of the isolates was performed by Kirby Bauer disc diffusion method following CLSI guidelines. S. pyogenes was isolated from 10.9% (38/350) of the screened children. The GAS colonisation rate was statistically insignificant (P>0.05) with sex and age sub-groups, although the rate was slightly higher among girls and age sub-group 9-12 years. No significant difference in carrier rate was observed among different schools (P>0.05). All isolates were susceptible to azithromycin. No resistance was detected for penicillin and its derivative antibiotic ampicillin. Highest resistance rate was observed for cotrimoxazole (71.0%) followed by chloramphenicol (7.8%), ciprofloxacin (5.2%) and erythromycin (5.2%). Antibiotic resistant GAS isolated from asymptomatic Nepalese school children is a public health concern. When screened and appropriately treated with antibiotics, carriers can be prevented from spreading of streptococcal infections in the school environment and the community. Preventing cross infections would ultimately reduce the incidence of life-threatening sequelae which are debilitating and difficult to treat. It is recommended to conduct regular screening and GAS surveillance in schools, and maintain rational use of antibiotics to minimise GAS carriage/infections and resistance.


Peripheral venous bicarbonate levels as a marker of predicting severity in acute pancreatitis: a retrospective study.
Acute pancreatitis is an inflammatory process of the pancreas, which can range from mild, self-limited disease to severe disease potentially resulting in death. Although overall mortality has decreased, the incidence of acute pancreatitis is increasing. Severe episodes of acute pancreatitis are more likely to result in prolonged hospitalisation and increased mortality. Reliable means of early detection and indicators of severity in acute pancreatitis remain a challenge, hence the continued efforts of the medical community toward developing improved prognostic tools. There are various scoring systems available that are aimed at classifying severity of acute pancreatitis; however, many of these scores are cumbersome and remain underutilised. As a result, the investigation of biological markers with potential to predict prognosis of acute pancreatitis has been a topic of interest. To investigate the utility of peripheral venous bicarbonate levels as a biomarker in predicting severity of acute pancreatitis, defined as increased length of stay, organ failure, need for intervention, and/or mortality. Patients between the ages of 18 and 80 who were admitted from September 2015 to August 2017 with acute pancreatitis were selected via chart review. The associations between peripheral venous bicarbonate level obtained on admission and length of stay, encounter type, organ failure and need for intervention were analyzed. There was a significant association between bicarbonate levels and both discharge type and organ failure. Expired patients and patients with more incidences of organ failure during their hospitalization were found to have lower peripheral venous bicarbonate levels on admission. There was no significant association found between bicarbonate level and length of stay or need for intervention. Our retrospective study found that lower peripheral venous bicarbonate levels were significantly associated with increased incidence of organ failure and mortality in acute pancreatitis. Peripheral venous sampling can be promptly and easily obtained in most clinical settings, making this biological marker worthy of further investigation.


Growth response to growth hormone (GH) treatment relates to serum insulin-like growth factor I (IGF-I) and IGF-binding protein-3 in short children with various GH secretion capacities. Swedish Study Group for Growth Hormone Treatment.
The purpose of the study was to evaluate the relationship between the 1-yr (n = 193) and 2-yr (n = 128) growth response and the individual serum concentrations of insulin-like growth factor I (IGF-I) and IGF-binding protein 3 (IGFBP-3) before and during GH treatment. Our study group of prepubertal short children had from very low to high GH secretory capacity, estimated during an arginine-insulin tolerance test, and the ages ranged from 3-15 yr at the start of treatment. Their serum levels of IGF-I and IGFBP-3 were low before treatment compared to those in an age-related reference group of prepubertal children and increased significantly from the start to 1 month of GH treatment. The mean increase in height SD score was 0.80 SD score after 1 yr of GH treatment and 1.26 SD score after 2 yr, with a wide range. In univariate analyses the highest correlation coefficients to the 2-yr growth response were found to be vs. the following variables from the start of treatment: IGF-I SD score (r = -0.49), log maximum GH concentration (log GHmax) during the arginine-insulin tolerance test (r = -0.47), difference between the height SD score of the individual child and the midparental height SD score (diffSD score; r = -0.45), IGFBP-3 SD score (r = -0.39), age (r = -0.30), short term change in IGFBP-3 SD score (r = 0.37), and IGF-I SD score (r = 0.34). In multivariate stepwise regression analysis, 41% of the variation in the 2-yr growth response could be explained by IGF-I SD score or log GHmax together with age at the start of treatment, weight SD score at 1 yr of age, and diffSD score. When both IGF-I SD score and GHmax were included and when the short term changes in IGF-I SD score were added, 46% and 58% of the variation, respectively, could be explained. The regression algorithms using different combinations of variables and their corresponding prediction intervals are also presented.


Quinine Ingestion During the Latter Stages of a 3,000-m Time Trial Fails to Improve Cycling Performance.
The ingestion of quinine, a bitter tastant, improves short-term (30 s) cycling performance, but it is unclear whether this effect can be integrated into the last effort of a longer race. The purpose of this study was to determine whether midtrial quinine ingestion improves 3,000-m cycling time-trial (TT) performance. Following three familiarization TTs, 12 well-trained male cyclists (mean ± SD: mass = 76.6 ± 9.2 kg, maximal aerobic power = 390 ± 50 W, maximal oxygen uptake = 4.7 ± 0.6 L/min) performed four experimental 3,000-m TTs on consecutive days. This double-blind, crossover design study had four randomized and counterbalanced conditions: (a) Quinine 1 (25-ml solution, 2 mM of quinine); (b) Quinine 2, replicate of Quinine 1; (c) a 25-ml sweet-tasting no-carbohydrate solution (Placebo); and (d) 25 ml of water (Control) consumed at the 1,850-m point of the TT. The participants completed a series of perceptual scales at the start and completion of all TTs, and the power output was monitored continuously throughout all trials. The power output for the last 1,000 m for all four conditions was similar: mean ± SD: Quinine 1 = 360 ± 63 W, Quinine 2 = 367 ± 63 W, Placebo = 364 ± 64 W, and Control = 367 ± 58 W. There were also no differences in the 3,000-m TT power output between conditions. The small perceptual differences between trials at specific 150-m splits were not explained by quinine intake. Ingesting 2 mM of quinine during the last stage of a 3,000-m TT did not improve cycling performance.


Characterization of a central Ca2+/calmodulin-dependent protein kinase IIalpha/beta binding domain in densin that selectively modulates glutamate receptor subunit phosphorylation.
The densin C-terminal domain can target Ca(2+)/calmodulin-dependent protein kinase IIα (CaMKIIα) in cells. Although the C-terminal domain selectively binds CaMKIIα in vitro, full-length densin associates with CaMKIIα or CaMKIIβ in brain extracts and in transfected HEK293 cells. This interaction requires a second central CaMKII binding site, the densin-IN domain, and an "open" activated CaMKII conformation caused by Ca(2+)/calmodulin binding, autophosphorylation at Thr-286/287, or mutation of Thr-286/287 to Asp. Mutations in the densin-IN domain (L815E) or in the CaMKIIα/β catalytic domain (I205/206K) disrupt the interaction. The amino acid sequence of the densin-IN domain is similar to the CaMKII inhibitor protein, CaMKIIN, and a CaMKIIN peptide competitively blocks CaMKII binding to densin. CaMKII is inhibited by both CaMKIIN and the densin-IN domain, but the inhibition by densin is substrate-selective. Phosphorylation of a model peptide substrate, syntide-2, or of Ser-831 in AMPA receptor GluA1 subunits is fully inhibited by densin. However, CaMKII phosphorylation of Ser-1303 in NMDA receptor GluN2B subunits is not effectively inhibited by densin in vitro or in intact cells. Thus, densin can target multiple CaMKII isoforms to differentially modulate phosphorylation of physiologically relevant downstream targets.


Effect of three countermeasures against the illegal crossing of railway tracks.
This study was designed to investigate the effects of three countermeasures--landscaping, building a fence and prohibitive signs--on the frequency of trespassing, which in this case means crossing the track at places where it is forbidden. At each location the official route was no more than 300 m away. The main results showed that the effect of each countermeasure on the frequency of trespassing was statistically significant. Specifically, the fencing reduced trespassing by 94.6%, followed by landscaping (91.3%) and prohibitive signs (30.7%). The majority of illegal crossings were committed alone and the persons trespassing were mostly adults and men. In addition, the results demonstrated some tendencies of how the effects of the selected countermeasures can vary with the characteristics of the trespassers. The main implication of this study is that the building of physical barriers such as landscaping or fencing is recommended for reducing trespassing. However, if the required resources are not available or the site is not suitable for such measures, the use of prohibitive signs is recommended. Further, there is a need to tailor the countermeasures to the characteristics of the trespassers in order to ensure that the most appropriate countermeasures are applied.


Remifentanil Reduces Blood Loss During Orthognathic Surgery.
Remifentanil is reported to reduce oral tissue blood flow. We performed a retrospective investigation using logistic regression analysis of anesthesia records to investigate whether the use of remifentanil infusion in a balanced anesthesia technique was useful as a primary technique to reduce blood loss during orthognathic surgery. Subjects were 80 patients who underwent Le Fort I osteotomy and sagittal split ramus osteotomy of the mandible. The variables included gender, age, weight, type of maintenance anesthetic, type and dose or infusion rate of opioid, mean systolic blood pressure (SBP-mean), coefficient of variation of systolic blood pressure (CVSBP) during surgery, mean heart rate (HR-mean), duration of surgery, total blood loss, volume of infusion used, amount of local anesthetic used, body temperature, and urine output. Gender, type of maintenance anesthetic, type of opioid, SBP-mean, CVSBP, HR-mean, and duration of surgery were used as candidates for independent variables. Logistic regression analysis was performed for the selected independent variables with the total blood loss as the dependent variable. The factors associated with the reduction of blood loss were the use of remifentanil (odds ratio, 3.112; 95% CI, 1.166-8.307; P = .023) and smaller CVSBP (odds ratio, 2.747; 95% CI, 1.07-7.053; P = .036). Use of remifentanil and smaller CVSBP were associated with a reduction of blood loss during orthognathic surgery.


Appendix vermiformis as a left pyelo-ureteral substitute in a 6-month-old girl with solitary kidney.
Extensive ureteral loss in early childhood is a rare but dramatic event. We present the case of a 6-month-old girl with a iatrogenic extensive pyelo-ureteral loss and solitary kidney. She successfully underwent left ureteral substitution using the appendix vermiformis. Left ureteral reconstruction using the appendix vermiformis is a feasible procedure even in very small children.


High-dose intraarterial verapamil in the treatment of cerebral vasospasm after aneurysmal subarachnoid hemorrhage.
Because oral calcium channel blockers appear to reduce the severity of cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH), interest in their application intraarterially has emerged for cases in which noninvasive means of alleviating vasospasm are unsuccessful. Studies to date have been limited to the administration of low intraarterial doses because of concerns about hemodynamic stability and changes in intracranial pressure. These doses, although effective in cases of milder vasospasm, were inadequate in severe cases. The authors present a series of 10 patients with cerebral vasospasm who underwent 12 procedures in which they received > or = 20 mg of intraarterial verapamil per procedure. A retrospective review was undertaken of all patients who underwent endovascular treatment for cerebral vasospasm due to aneurysmal SAH by the senior author between February 2005 and October 2006. Ten patients were identified who had undergone a total of 12 procedures during which > or =20 mg of intraarterial verapamil had been administered. From angiography reports, anesthesia records, and nursing records, we obtained pre- and postverapamil mean arterial blood pressures (MABPs), heart rates, intracranial pressures (ICPs) (when available), and visible changes in the degree of vasospasm. No statistically significant changes in MABP, heart rate, or ICP were observed after administration of > or = 20 mg of intraarterial verapamil, and the degree of improvement in vasospasm was statistically significant based on our grading system. No correlation was found between the change in hemodynamic parameters and the total dose of verapamil. This study indicates that high-dose intraarterial verapamil may be used to treat cerebral vasospasm without compromising hemodynamic stability or increasing ICP.


Bacteroides gracilis, an important anaerobic bacterial pathogen.
Clinical isolates of agar-pitting, formate-fumarate-requiring, anaerobic gram-negative bacilli were recharacterized in consideration of the species descriptions of Bacteroides ureolyticus and the newly described B. gracilis, Campylobacter concisus, and Wolinella sp. During an 11-year period, 7.5% (101 of 1,341) of all specimens yielding anaerobes were found to contain an organism in this group. Of the 71 isolates that were available for study, 43 were B. ureolyticus, 23 were B. gracilis, and 5 were in the Wolinella-C. concisus group. The role in infection and patterns of antimicrobial susceptibility for B. ureolyticus and B. gracilis were studied. Review of the clinical data indicated that 83% of B. gracilis strains were recovered from patients with serious visceral or head and neck infection, whereas 73% of B. ureolyticus isolates were recovered from superficial soft-tissue or bone infections. The strains of B. ureolyticus were uniformly susceptible to the tested antimicrobial agents. B. gracilis, however, showed some striking resistance, with penicillin active against only 67% and the cephalosporins active against 67 to 84% of the isolates tested. The association of B. gracilis with serious deep-tissue infection, coupled with the relatively high frequency of antibiotic resistance, indicates that it is an important, previously unrecognized, pathogen.


Heat flows in rock cracks naturally optimize salt compositions for ribozymes.
Catalytic nucleic acids, such as ribozymes, are central to a variety of origin-of-life scenarios. Typically, they require elevated magnesium concentrations for folding and activity, but their function can be inhibited by high concentrations of monovalent salts. Here we show that geologically plausible high-sodium, low-magnesium solutions derived from leaching basalt (rock and remelted glass) inhibit ribozyme catalysis, but that this activity can be rescued by selective magnesium up-concentration by heat flow across rock fissures. In contrast to up-concentration by dehydration or freezing, this system is so far from equilibrium that it can actively alter the Mg:Na salt ratio to an extent that enables key ribozyme activities, such as self-replication and RNA extension, in otherwise challenging solution conditions. The principle demonstrated here is applicable to a broad range of salt concentrations and compositions, and, as such, highly relevant to various origin-of-life scenarios.