Surface Engineering of FLT4-Targeted Nanocarriers Enhances Cell-Softening Glaucoma Therapy.
Primary open-angle glaucoma is associated with elevated intraocular pressure (IOP) that damages the optic nerve and leads to gradual vision loss. Several agents that reduce the stiffness of pressure-regulating Schlemm's canal (SC) endothelial cells, in the conventional outflow pathway of the eye, lower IOP in glaucoma patients and are approved for clinical use. However, poor drug penetration and uncontrolled biodistribution limit their efficacy and produce local adverse effects. Compared to other ocular endothelia, FLT4/VEGFR3 is expressed at elevated levels by SC endothelial cells and can be exploited for targeted drug delivery. Here, we validate FLT4 receptors as clinically relevant targets on SC cells from glaucomatous human donors and engineer polymeric self-assembled nanocarriers displaying lipid-anchored targeting ligands that optimally engage this receptor. Targeting constructs were synthesized as lipid-PEGx-peptide, differing in the number of PEG spacer units (x), and were embedded in micelles. We present a novel proteolysis assay for quantifying ligand accessibility that we employ to design and optimize our FLT4-targeting strategy for glaucoma nanotherapy. Peptide accessibility to proteases correlated with receptor-mediated targeting enhancements. Increasing the accessibility of FLT4-binding peptides enhanced nanocarrier uptake by SC cells while simultaneously decreasing the uptake by off-target vascular endothelial cells. Using a paired longitudinal IOP study in vivo, we show that this enhanced targeting of SC cells translates to IOP reductions that are sustained for a significantly longer time as compared to controls. Confocal microscopy of murine anterior segment tissue confirmed nanocarrier localization to SC within 1 h after intracameral administration. This work demonstrates that steric effects between surface-displayed ligands and PEG coronas significantly impact the targeting performance of synthetic nanocarriers across multiple biological scales. Minimizing the obstruction of modular targeting ligands by PEG measurably improved the efficacy of glaucoma nanotherapy and is an important consideration for engineering PEGylated nanocarriers for targeted drug delivery.


Clinical features of cardiac nodularity-like appearance induced by Helicobacter pylori infection.
We have previously reported that Helicobacter pylori (H. pylori)-associated nodular gastritis could occur in both the antrum and the cardia. Cardiac nodularity-like appearance (hereafter, called as cardiac nodularity) had a high predictive accuracy for the diagnosis of H. pylori infection. In the previous study, we included only the patients who were evaluated for H. pylori infection for the first time, and excluded patients with a history of eradication. Therefore, the prevalence and clinical features of cardiac nodularity remains unknown. To perform this cross-sectional study to explore the characteristics of cardiac nodularity. Consecutive patients who underwent esophagogastroduodenoscopy between May, 2017 and August, 2019 in the Toyoshima Endoscopy Clinic were enrolled in this study. We included H. pylori-negative, H. pylori-positive, and H. pylori-eradicated patients, and excluded patients with unclear H. pylori status and eradication failure. H. pylori infection was diagnosed according to serum anti-H. pylori antibody and the urea breath test or histology. Cardiac nodularity was defined as a miliary nodular appearance or the presence of scattered whitish circular small colorations within 2 cm of the esophagogastric junction. Nodularity was visualized as whitish in the narrow-band imaging mode. We collected data on the patients' baseline characteristics. A total of 1078 patients were finally included. Among H. pylori-negative patients, cardiac nodularity and antral nodularity were recognized in 0.14% each. Among H. pylori-positive patients, cardiac nodularity and antral nodularity were recognized in 54.5% and 29.5%, respectively. Among H. pylori-eradicated patients, cardiac nodularity and antral nodularity were recognized in 4.5% and 0.6%, respectively. The frequency of cardiac nodularity was significantly higher than that of antral nodularity in H. pylori-positive and -eradicated patients. The frequencies of cardiac nodularity and antral nodularity in H. pylori-eradicated patients were significantly lower than those in H. pylori-positive patients (P < 0.001). The patients with cardiac nodularity were significantly younger than those without cardiac nodularity (P = 0.0013). Intestinal metaplasia score of the patients with cardiac nodularity were significantly lower than those without cardiac nodularity (P = 0.0216). Among H. pylori-eradicated patients, the patients with cardiac nodularity underwent eradication significantly more recently compared with those without cardiac nodularity (P < 0.0001). This report outlines the prevalence and clinical features of cardiac nodularity, and confirm its close association with active H. pylori infection.


The ubiquinone-binding site of the Saccharomyces cerevisiae succinate-ubiquinone oxidoreductase is a source of superoxide.
The mitochondrial succinate dehydrogenase (SDH) is a tetrameric iron-sulfur flavoprotein of the Krebs cycle and of the respiratory chain. A number of mutations in human SDH genes are responsible for the development of paragangliomas, cancers of the head and neck region. The mev-1 mutation in the Caenorhabditis elegans gene encoding the homolog of the SDHC subunit results in premature aging and hypersensitivity to oxidative stress. It also increases the production of superoxide radicals by the enzyme. In this work, we used the yeast succinate dehydrogenase to investigate the molecular and catalytic effects of paraganglioma- and mev-1-like mutations. We mutated Pro-190 of the yeast Sdh2p subunit to Gln (P190Q) and recreated the C. elegans mev-1 mutation by converting Ser-94 in the Sdh3p subunit into a glutamate residue (S94E). The P190Q and S94E mutants have reduced succinate-ubiquinone oxidoreductase activities and are hypersensitive to oxygen and paraquat. Although the mutant enzymes have lower turnover numbers for ubiquinol reduction, larger fractions of the remaining activities are diverted toward superoxide production. The P190Q and S94E mutations are located near the proximal ubiquinone-binding site, suggesting that the superoxide radicals may originate from a ubisemiquinone intermediate formed at this site during the catalytic cycle. We suggest that certain mutations in SDH can make it a significant source of superoxide production in mitochondria, which may contribute directly to disease progression. Our data also challenge the dogma that superoxide production by SDH is a flavin-mediated event rather than a quinone-mediated one.


The educational gradient in cardiovascular risk factors: impact of shared family factors in 228,346 Norwegian siblings.
Various indicators of childhood socioeconomic position have been related to cardiovascular disease (CVD) risk in adulthood. We investigated the impact of shared family factors on the educational gradient in midlife CVD risk factors by assessing within sibling similarities in the gradient using a discordant sibling design. Norwegian health survey data (1980-2003) was linked to educational and generational data. Participants with a full sibling in the health surveys (228,346 individuals in 98,046 sibships) were included. Associations between attained educational level (7-9 years, 10-11 years, 12 years, 13-16 years, or >16 years) and CVD risk factor levels in the study population was compared with the corresponding associations within siblings. Educational gradients in risk factors were attenuated when factors shared by siblings was taken into account: A one category lower educational level was associated with 0.7 (95% confidence interval 0.6 to 0.8) mm Hg higher systolic blood pressure (27% attenuation), 0.4 (0.4 to 0.5) mmHg higher diastolic blood pressure (30%), 1.0 (1.0 to 1.1) more beats per minute higher heart rate (21%), 0.07 (0.06 to 0.07) mmol/l higher serum total cholesterol (32%), 0.2 (0.2 to 0.2) higher smoking level (5 categories) (30%), 0.15 (0.13 to 0.17) kg/m2 higher BMI (43%), and 0.2 (0.2 to 0.2) cm lower height (52%). Attenuation increased with shorter age-difference between siblings. About one third of the educational gradients in modifiable CVD risk factors may be explained by factors that siblings share. This implies that childhood environment is important for the prevention of CVD.


Melatonin levels decrease in type 2 diabetic patients with cardiac autonomic neuropathy.
The present study has been designed to determine melatonin levels in type 2 diabetic patients and test the relationship between the autonomic nervous system and melatonin dynamics. Thirty-six type 2 diabetic patients and 13 age-matched healthy subjects were recruited for the study. Circadian rhythm of melatonin secretion was assessed by measuring serum melatonin concentrations between 02:00-04:00 and 16:00-18:00 hr. Melatonin dynamics were re-evaluated with respect to autonomic nervous system in diabetic patients with autonomic neuropathy who were diagnosed by the cardiovascular reflex tests, heart rate variability (HRV), and 24-hr blood pressure monitoring. Nocturnal melatonin levels and the nocturnal melatonin surge were low in the diabetic group (P = 0.027 and 0.008 respectively). Patients with autonomic neuropathy revealed decreased melatonin levels both at night and during day when compared with healthy controls (P < 0.001 and 0.004 respectively) while the melatonin dynamics were similar to controls in patients without autonomic neuropathy. Nocturnal melatonin level was positively correlated with nocturnal high and low frequency components of HRV (P = 0.005 and 0.011 respectively) and systolic and diastolic blood pressures at night (P = 0.002 and 0.004 respectively) in patients with autonomic neuropathy. We found a negative correlation between nocturnal melatonin levels and the degree of systolic blood pressure decrease at night (r = -0.478, P = 0.045). As a conclusion this study has shown that circadian rhythm of melatonin secretion is blunted in type 2 diabetic patients and there is a complex relationship between various components of autonomic nervous system and melatonin secretion at night. Among the patients with autonomic neuropathy those with more preserved HRV and the systolic nondippers (<10% reduction in blood pressure during the night relative to daytime values) have more pronounced melatonin surge at night.


The effect of childhood trauma on blood transcriptome expression in major depressive disorder.
Childhood trauma (CT) increases the likelihood of developing severe mental illnesses, such as major depressive disorder (MDD), during adulthood. Several studies have suggested an inflammatory immune system dysregulation as a biological mediator; however, the molecular mechanisms underlying this relationship remain largely undetermined. Moreover, different types of CT, in particular, emotional abuse and neglect, confer a higher risk of developing MDD, and recent meta-analyses showed that each CT can be associated with different pro-inflammatory biomarkers. However, no studies using a hypothesis-free approach have been performed. For this reason, we carried out a reanalysis of transcriptome data from a large mRNA sequencing dataset to investigate different types of CT in MDD patients. Gene expression analysis followed by principal component and gene-set enrichment analyses were carried out to identify genes and pathways differentially expressed in 368 patients who experienced four different types of CT (sexual abuse, physical abuse, emotional abuse and neglect). Expression analysis of single genes revealed a significant association between the neglect CT and the MED22 gene (p = 1.11 × 10-6; FDR = 0.016). Furthermore, analyses of the principal components of expression data support a dysregulation of cytokine system pathways, such as interferon (IFN) α/β and γ signaling, as a consequence of emotional abuse in depressed patients. Our results corroborate the hypothesis that specific types of CT affect distinct molecular pathways, and in particular, emotional abuse and neglect exert the strongest impact on gene expression in MDD.


The impact of household air cleaners on the oxidative potential of PM2.5 and the role of metals and sources associated with indoor and outdoor exposure.
The health effects associated with human exposure to airborne fine particulate matter (PM2.5) have been linked to the ability of PM2.5 to facilitate the production of excess cellular reactive oxygen species (oxidative potential). Concern about the adverse human health impacts of PM2.5 has led to the increased use of indoor air cleaners to improve indoor air quality, which can be an important environment for PM2.5 exposure. However, the degree to which the oxidative potential of indoor and personal PM2.5 can be influenced by an indoor air cleaner remains unclear. In this study we enrolled 43 children with physician diagnosed asthma in suburban Shanghai, China and collected two paired-sets of 48-h indoor, outdoor, and personal PM2.5 exposure samples. One set of samples was collected under "real filtration" during which a functioning air cleaner was installed in the child's bedroom, and the other ("false filtration") with an air cleaner without internal filters. The PM2.5 samples were characterized by inductively coupled plasma mass spectroscopy for elements, and by an alveolar macrophage assay for oxidative potential. The sources of metals contributing to our samples were determined by the EPA Positive Matrix Factorization model. The oxidative potential was lower under real filtration compared to sham for indoor (median real/sham ratio: 0.260) and personal exposure (0.813) samples. Additionally, the sources of elements in PM2.5 that were reduced indoors and personal exposure samples by the air cleaner (e.g. regional aerosol and roadway emissions) were found by univariate multiple regression models to be among those contributing to the oxidative potential of the samples. An IQR increase in the regional aerosol and roadway emissions sources was associated with a 107% (95% CI: 80.1-138%) and 38.1% (17.6-62.1%) increase in measured oxidative potential respectively. Our results indicate that indoor air cleaners can reduce the oxidative potential of indoor and personal exposure to PM2.5, which may lead to improved human health.


The cardiokine secreted Frizzled-related protein 3, a modulator of Wnt signalling, in clinical and experimental heart failure.
Experimental studies have shown involvement of Wnt signalling in heart failure (HF). We hypothesized that secreted frizzled-related protein 3 (sFRP3), a modulator of Wnt signalling, is related to the progression of HF. Circulating sFRP3 was measured in 153 HF patients and compared with 25 healthy controls. The association of sFRP3 with mortality was evaluated in 1202 patients (GISSI-HF trial). sFRP3 mRNA expression was assessed in failing human and murine left ventricles (LV), and cellular localization was determined after fractioning of myocardial tissue. In vitro studies were carried out in cardiac fibroblasts subjected to cyclic mechanical stretch. (i) Heart failure patients had significantly raised serum sFRP3 levels compared with controls, (ii) during a median follow-up of 47 months, 315 patients died in the GISSI-HF substudy. In univariable Cox regression, tertiles of baseline sFRP3 concentration were significantly associated with all-cause and cardiovascular mortality. After adjustment for demographic and clinical variables, but not for CRP and NT-proBNP, the associations with mortality remained significant for the third tertile (all-cause, HR 1.45, P = 0.011; cardiovascular, HR 1.66, P = 0.003), (iii) sFRP3 mRNA expression was increased in failing human LV, with a decline following LV assist device therapy. LV from post-MI mice showed an increased sFRP3 mRNA level, particularly in cardiac fibroblasts, and (iv) mechanical stretch enhanced sFRP3 expression and release in myocardial fibroblasts. There is an association between increased sFRP3 expression and adverse outcome in HF, suggesting that the failing myocardium itself contributes to an increase in circulating sFRP3.


Neutrophils responsive to endogenous IFN-beta regulate tumor angiogenesis and growth in a mouse tumor model.
Angiogenesis is a hallmark of malignant neoplasias, as the formation of new blood vessels is required for tumors to acquire oxygen and nutrients essential for their continued growth and metastasis. However, the signaling pathways leading to tumor vascularization are not fully understood. Here, using a transplantable mouse tumor model, we have demonstrated that endogenous IFN-beta inhibits tumor angiogenesis through repression of genes encoding proangiogenic and homing factors in tumor-infiltrating neutrophils. We determined that IFN-beta-deficient mice injected with B16F10 melanoma or MCA205 fibrosarcoma cells developed faster-growing tumors with better-developed blood vessels than did syngeneic control mice. These tumors displayed enhanced infiltration by CD11b+Gr1+ neutrophils expressing elevated levels of the genes encoding the proangiogenic factors VEGF and MMP9 and the homing receptor CXCR4. They also expressed higher levels of the transcription factors c-myc and STAT3, known regulators of VEGF, MMP9, and CXCR4. In vitro, treatment of these tumor-infiltrating neutrophils with low levels of IFN-beta restored expression of proangiogenic factors to control levels. Moreover, depletion of these neutrophils inhibited tumor growth in both control and IFN-beta-deficient mice. We therefore suggest that constitutively produced endogenous IFN-beta is an important mediator of innate tumor surveillance. Further, we believe our data help to explain the therapeutic effect of IFN treatment during the early stages of cancer development.


Proof-of-principle investigation of an algorithmic model of adenosine-mediated angiogenesis.
We investigated an algorithmic approach to modelling angiogenesis controlled by vascular endothelial growth factor (VEGF), the anti-angiogenic soluble VEGF receptor 1 (sVEGFR-1) and adenosine (Ado). We explored its feasibility to test angiogenesis-relevant hypotheses. We illustrated its potential to investigate the role of Ado as an angiogenesis modulator by enhancing VEGF activity and antagonizing sVEGFR-1. We implemented an algorithmic model of angiogenesis consisting of the dynamic interaction of endothelial cells, VEGF, sVEGFR-1 and Ado entities. The model is based on a logic rule-based methodology in which the local behaviour of the cells and molecules is encoded using if-then rules. The model shows how Ado may enhance angiogenesis through activating and inhibiting effects on VEGF and sVEGFR-1 respectively. Despite the relative simplicity of the model, it recapitulated basic features observed in in vitro models. However, observed disagreements between our models and in vitro data suggest possible knowledge gaps and may guide future experimental directions. The proposed model can support the exploration of hypotheses about the role of different molecular entities and experimental conditions in angiogenesis. Future expansions can also be applied to assist research planning in this and other biomedical domains.


Release of hormones from conjugates: chloroplast expression of β-glucosidase results in elevated phytohormone levels associated with significant increase in biomass and protection from aphids or whiteflies conferred by sucrose esters.
Transplastomic tobacco (Nicotiana tabacum) plants expressing β-glucosidase (Bgl-1) show modified development. They flower 1 month earlier with an increase in biomass (1.9-fold), height (1.5-fold), and leaf area (1.6-fold) than untransformed plants. Trichome density on the upper and lower leaf surfaces of BGL-1 plants increase by 10- and 7-fold, respectively, harboring 5-fold more glandular trichomes (as determined by rhodamine B staining), suggesting that BGL-1 lines produce more sugar esters than control plants. Gibberellin (GA) levels were investigated because it is a known regulator of flowering time, plant height, and trichome development. Both GA(1) and GA(4) levels are 2-fold higher in BGL-1 leaves than in untransformed plants but do not increase in other organs. In addition, elevated levels of other plant hormones, including zeatin and indole-3-acetic acid, are observed in BGL-1 lines. Protoplasts from BGL-1 lines divide and form calli without exogenous hormones. Cell division in protoplasts is enhanced 7-fold in the presence of exogenously applied zeatin-O-glucoside conjugate, indicating the release of active hormones from their conjugates. Whitefly (Bemisia tabaci) and aphid (Myzus persicae) populations in control plants are 18 and 15 times higher than in transplastomic lines, respectively. Lethal dose to kill 50% of the test population values of 26.3 and 39.2 μg per whitefly and 23.1 and 35.2 μg per aphid for BGL-1 and untransformed control exudates, respectively, confirm the enhanced toxicity of transplastomic exudates. These data indicate that increase in sugar ester levels in BGL-1 lines might function as an effective biopesticide. This study provides a novel strategy for designing plants for enhanced biomass production and insect control by releasing plant hormones or sugar esters from their conjugates stored within their chloroplasts.


2D Image Quantification of Microbial Iron Chelators (Siderophores) Using Diffusive Equilibrium in Thin Films Method.
Siderophores are natural metal chelating agents that strongly control the biogeochemical metal cycles such as Fe in the environment. This article describes a new methodology to detect and quantify at the micromolar concentration the spatial distribution at millimeter scale of siderophores within the root's system. The "universal" CAS assay originally designed for bacterial siderophores detection and later designed for fungus was adapted here for diffusive equilibrium in thin film gel techniques (DET). The method was calibrated against the marketed desferrioxamine mesylate (DFOM) siderophore and applied with experiments performed with sunflower ( Helianthus annuus) and wheat ( Triticum aestivum) cultivated on free iron agar medium plates. We present here the first results with 2D images of the siderophores distribution in the vicinity of the root system of plants. With this technique we detected (i) the production of siderophores on bacteria inoculated ( Pseudomonas fluorescens) environments and (ii) hotspots of natural iron binding ligands production up to 50 μM in the wheat rhizosphere. The lower detection limit in our experiment was 2.5 μmol/L. This new technique offers a unique opportunity to investigate the siderophore production in two dimensions in a wide range of applications from laboratory experiments to natural systems very likely using an in situ and nondestructive tool.


Messengers of cell death: apoptotic signaling in health and disease.
Apoptosis is a genetically controlled mechanism of cell death involved in the regulation of tissue homeostasis. Understanding the molecular basis of apoptosis signaling may reveal novel clues for lymphomagenesis. Pro-apoptotic signaling is mediated by specific ligands and surface death receptors (extrinsic pathway of apoptosis regulation), which are capable of delivering a death signal from the microenvironment and can activate the execution of apoptosis in the cell cytoplasm and organelles. Death receptors include tumor necrosis factor-receptor 1, Fas, death receptor (DR) 3, DR4, DR5 and DR6, whereas death ligands include tumor necrosis factor-a, lymphotoxin, Fas-Ligand, Apo3-Ligand and TRAIL (TNF-Related Apoptosis-Inducing Ligand). Once activated, death receptors recruit adaptor proteins, which in turn recruit initiator caspases, giving rise to a pro-apoptotic complex termed the death-inducing signaling complex (DISC). Besides being triggered from microenvironmental signals, apoptosis can also be activated from inside the cell through specific cell sensors residing in the cell nucleus and cytoplasm (intrinsic pathway of apoptosis regulation). The intrinsic pathway of apoptosis leads to the formation of a pro-apoptotic complex termed an apoptosome. Both the extrinsic and the intrinsic pathways of apoptosis signaling converge into a common pathway causing the activation of the effector enzymes caspases. Consistent with the role of apoptosis as a main regulator of B-cell homeostasis in the germinal center, the pathogenesis of several germinal-center-derived lymphomas is characterized by deregulation of one or more steps of the apoptosis signaling pathways. Tumor-specific alterations in the apoptotic machinery may represent new potential targets for molecular therapy of lymphoma.


A combination of antigenic regions of Toxoplasma gondii microneme proteins induces protective immunity against oral infection with parasite cysts.
Infection with Toxoplasma gondii causes morbidity and mortality in congenitally infected and immunocompromised individuals. Both humoral and cell-mediated immunity are involved in host resistance to invasion of the parasite. Among putative vaccine candidates, the T. gondii microneme proteins appear to be promising, because they are responsible for the invasion process. The present work focused on studying the immunogenicity of microneme proteins in infected individuals and in a mouse model of chronic toxoplasmosis. We identified 5 distinct antigenic regions within MIC2, MIC4, MIC2-associated protein, and apical membrane antigen 1 gene products, which were recognized by (1) T cells from both adults with acquired infection and children with congenital infection and (2) antibodies from all patients. Finally, we demonstrated that DNA immunization with microneme fragments elicited effective protection in mice (84% reduction in brain-cyst burden), suggesting that a combination of these antigenic regions should be considered in the design of potential vaccines against toxoplasmosis.


Alternative methods to visual and radiographic examinations for approximal caries detection.
A shift in caries prevalence from occlusal surfaces to approximal surfaces has been demonstrated by epidemiological studies. Two recent meta-analyses evaluated the performance of visual examination and radiography for carious lesion detection, and reported low sensitivity but high specificity for early approximal caries detection. This suggests that the conventional methods have a higher risk of failing to detect approximal lesions. Consequently, in caries susceptible populations, there is a risk of progression of non-cavitated lesions to irreversible tooth destruction before the lesions are detected. This paper aims to review the performance of unconventional and novel methods for approximal caries detection. In vitro and in vivo studies identified through a MEDLINE search using keywords such as caries detection, approximal caries detection, light fluorescence and dental caries, and transillumination and dental caries were reviewed. The unconventional methods known to be used for approximal caries detection and included in this review are: cone beam computed tomography, fiber-optic trans-illumination, digital imaging fiber optic trans-illumination, near-infrared digital imaging transillumination, optical coherence tomography, laser fluorescence, ultrasound, and LED reflection and refraction.


Distinct and overlapping patterns of cytokine regulation of thymic and bone marrow-derived NK cell development.
Although bone marrow (BM) represents the main site for postnatal NK cell development, recently a distinct thymic-dependent NK cell pathway was identified. These studies were designed to investigate the role of cytokines in regulation of thymic NK cells and to compare with established regulatory pathways of BM-dependent NK cell compartment. The common cytokine receptor gamma-chain (Il2rg) essential for IL-15-induced signaling, and FMS-like tyrosine kinase 3 (FLT3) receptor ligand (Flt3l) were previously identified as important regulatory pathways of the BM NK cell compartment based on lack of function studies in mice, however their complementary action remains unknown. By investigating mice double-deficient in Il2rg and Flt3l (Flt3l(-/-) Il2rg(-/-)), we demonstrate that FLT3L is important for IL2Rg-independent maintenance of both immature BM as well as peripheral NK cells. In contrast to IL-7, which is dispensable for BM but important for thymic NK cells, IL-15 has a direct and important role in both thymic and BM NK cell compartments. Although thymic NK cells were not affected in Flt3l(-/-) mice, Flt3l(-/-)Il2rg(-/-) mice lacked detectable thymic NK cells, suggesting that FLT3L is also important for IL-2Rg-independent maintenance of thymic NK cells. Thus, IL-2Rg cytokines and FLT3L play complementary roles and are indispensable for homeostasis of both BM and thymic dependent NK cell development, suggesting that the cytokine pathways crucial for these two distinct NK cell pathways are largely overlapping.


Needle thoracentesis decompression: observations from postmortem computed tomography and autopsy.
Needle thoracentesis decompression (NTD) is a recommended emergency treatment for tension pneumothorax. Current doctrine recognizes two suitable sites: the second intercostal space in the midclavicular line and the fourth or fifth intercostal space in the anterior axillary line. A review was conducted of postmortem computed tomography and autopsy results in 16 cases where NTD was performed as an emergency procedure. In 16 cases with 23 attempted procedures, the outcome was confirmed in 17 attempts. In 7 placements, the catheter was in the pleural cavity; in 7 placements, the catheter never entered the pleural cavity; and in 3 placements, cavity penetration was verified at autopsy even though the catheter was no longer in the cavity. Success was noted in 6 of 13 anterior attempts and 4 of 4 lateral attempts, for an overall success rate of 59% (10 of 17). In the remaining 6 attempted procedures, a catheter was noted in the soft tissue on imaging; however, presence or absence of pleural cavity penetration was equivocal. All placements were attempted in the combat environment; no information is available about specifically where or by whom. NTD via a lateral approach was more successful than that via an anterior approach, although it was used in fewer cases. This supports the revision of the Tactical Combat Casualty Care Guidelines specifying the lateral approach as an alternative to an anterior approach.


Possible link between Toxoplasma gondii and the anosmia associated with neurodegenerative diseases.
Toxoplasma gondii is an intracellular protozoan infecting 30% to 50% of global human population. Recently, it was suggested that chronic latent neuroinflammation caused by the parasite may be responsible for the development of several neurodegenerative diseases manifesting with the loss of smell. Studies in animals inoculated with the parasite revealed cysts in various regions of the brain, including olfactory bulb. Development of behavioral changes was paralleled by the preferential persistence of cysts in defined anatomic structures of the brain, depending on the host, strain of the parasite, its virulence, and route of inoculation. Olfactory dysfunction reported in Alzheimer's disease, multiple sclerosis, and schizophrenia was frequently associated with the significantly increased serum anti-T gondii immunoglobulin G antibody levels. Damage of the olfactory system may be also at least in part responsible for the development of depression because T gondii infection worsened mood in such patients, and the olfactory bulbectomized rat serves as a model of depression.


Effect of transrespiratory pressure on PETCO2-PaCO2 and ventilatory reflexes in humans.
Inspiratory muscle activity increases when lung volume is increased by continuous positive-pressure breathing in conscious human subjects (Green et al., Respir. Physiol. 35: 283-300, 1978). Because end-tidal CO2 pressure (PETCO2) does not change, these increases have not been attributed to chemoreflexes. However, continuous positive-pressure breathing at 20 cmH2O influences the end-tidal to arterial CO2 pressure differences (Folkow and Pappenheimer, J. Appl. Physiol. 8: 102-110, 1955). We have compared PETCO2 with arterial CO2 pressure (PaCO2). We have compared PETCO2 with arterial CO2 pressure (PaCO2) in healthy human subjects exposed to continuous positive airway pressure (10 cmH2O) or continuous negative pressure around the torso (-15 cmH2O) sufficient to increase mean lung volume by about 650 ml. The difference between PETCO2 and PaCO2 was not decreased, and we conclude that PETCO2 is a valid measure of chemical drive to ventilation in such circumstances. We observed substantial increases in respiratory muscle electromyograms during pressure breathing as seen previously and conclude this response must originate by proprioception. On average, the compensation of tidal volume thus afforded was complete, but the wide variability of individual responses suggests that there was a large cerebral cortical component in the responses seen here.


Pollution breaks down the genetic architecture of life history traits in Caenorhabditis elegans.
When pollution occurs in an environment, populations present suffer numerous negative and immediate effects on their life history traits. Their evolutionary potential to live in a highly stressful environment will depend on the selection pressure strengths and on the genetic structure, the trait heritability, and the genetic correlations between them. If expression of this structure changes in a stressful environment, it becomes necessary to quantify these changes to estimate the evolutionary potential of the population in this new environment. We studied the genetic structure for survival, fecundity, and early and late growth in isogenic lines of a Caenorhabditis elegans population subject to three different environments: a control environment, an environment polluted with uranium, and a high salt concentration environment. We found a heritability decrease in the polluted environments for fecundity and early growth, two traits that were the most heritable in the control environment. The genetic structure of the traits was particularly affected in the uranium polluted environment, probably due to generally low heritability in this environment. This could prevent selection from acting on traits despite the strong selection pressures exerted on them. Moreover, phenotypic traits were more strongly affected in the salt than in the uranium environment and the heritabilities were also lower in the latter environment. Consequently the decrease in heritability was not proportional to the population fitness reduction in the polluted environments. Our results suggest that pollution can alter the genetic structure of a C. elegans population, and thus modify its evolutionary potential.


Interactions of corneal cells with transforming growth factor beta 2-modified poly dimethyl siloxane surfaces.
The downgrowth of corneal epithelial cells at the interface of an artificial cornea and the host eye tissue poses a significant problem to be overcome in developing a successful implant. As a means of inhibiting the proliferation of corneal epithelial cells on the stromal surface of the implant, we examined the immobilization of transforming growth factor beta-2 (TGF-beta2) via a bifunctional poly ethylene glycol (PEG) spacer to poly dimethyl siloxane (PDMS) surfaces. Growth factor immobilization was confirmed by modification with (125)I-labeled TGF-beta 2. The modified surfaces were also characterized by advancing water contact angles, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Although the amount of growth factor covalently bound to the surface was difficult to quantify apparently due to strong interactions between the growth factor and the PEG layer and high levels of adsorption, differences in the modified surfaces, suggestive of the presence of a significant amount of TGF-beta 2, were found. In vitro interactions of the modified surfaces with human corneal epithelial and stromal cells were examined. Growth factor surface concentrations as well as culture in the absence and presence of serum and other adhesive proteins were examined. Corneal stromal and epithelial cells cultured on the TGF-beta 2-modified surfaces consistently gave results opposite to those expected. Likely, the most notable and surprising result was the almost complete lack of adhesion of the stromal cells, with coverage averaging between 3 and 5%. In comparison, corneal epithelial cell growth appeared to be promoted by the presence of the immobilized growth factor, with cell coverage averaging 50-60% at 7 days of culture. A TGF-beta 2 concentration effect was noted with both cell types in the absence of serum, with increases in the coverage at higher TGF-beta 2 concentrations. The observed cell growth appeared to be the result of interactions between the cells and active growth factor, because the addition of anti-TGF-beta 2 to the culture medium reduced cell coverage to levels similar to those noted on control surfaces. Therefore, although TGF-beta 2-modified surfaces may not be suitable as corneal epithelial cell inhibiting surfaces, interactions of surface immobilized growth factor and corneal cells are complex and should be further examined.


Adherence of Candida albicans to glow-discharge modified acrylic denture base polymers.
An important aetiologic factor in the pathogenesis of denture-induced stomatitis, is the presence of numerous yeasts, usually Candida albicans, on the fitting surfaces of dentures. In the present study, effect of glow-discharge plasma, a technique applied to increase surface wettability of acrylic resins, on candidial adherence was evaluated. The durability of glow-discharge modification with saliva coating was also evaluated. Samples including control and experimental groups were prepared by using heat compression mould technique. To create a hydrophobicity gradient, experimental groups were exposed to a radiofrequency glow discharge in an O2 atmosphere under different discharge powers. To characterize the wetting properties, an expression of surface hydrophobicity, contact angle measurements were performed by the sessile drop method. The organism used was C. albicans (ATTC10321). Acrylic samples were coated with unstimulated whole saliva collected from a healthy man. The fungal suspension was poured on saliva-inoculated samples and incubated at 37 degrees for 2 h. The samples were then fixed with glutaraldehyde and Gram stained. Adhered candidial cells were examined by light microscope. Diffuse Reflectance FTIR (DRIFT) and scanning electron-microscope examinations were also performed to evaluate the surface composition and roughness of the test groups. Glow-discharge plasma was found to be an effective means of increasing surface wettability even with salivary pellicle. Amounts of candida cells adhered were significantly higher in all the plasma treated surfaces than the unmodified control group (P < 0.001). It was concluded that improving the surface wettability of acrylic resins by glow-discharge plasma in O2 atmosphere increased the adherence of the C. albicans.


STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling.
The cellular innate immune system is essential for recognizing pathogen infection and for establishing effective host defence. But critical molecular determinants responsible for facilitating an appropriate immune response-following infection with DNA and RNA viruses, for example-remain to be identified. Here we report the identification, following expression cloning, of a molecule (STING; stimulator of interferon genes) that appears essential for effective innate immune signalling processes. It comprises five putative transmembrane regions, predominantly resides in the endoplasmic reticulum and is able to activate both NF-kappaB and IRF3 transcription pathways to induce expression of type I interferon (IFN-alpha and IFN-beta ) and exert a potent anti-viral state following expression. In contrast, loss of STING rendered murine embryonic fibroblasts extremely susceptible to negative-stranded virus infection, including vesicular stomatitis virus. Further, STING ablation abrogated the ability of intracellular B-form DNA, as well as members of the herpesvirus family, to induce IFN-beta, but did not significantly affect the Toll-like receptor (TLR) pathway. Yeast two-hybrid and co-immunoprecipitation studies indicated that STING interacts with RIG-I and with SSR2 (also known as TRAPbeta), which is a member of the translocon-associated protein (TRAP) complex required for protein translocation across the endoplasmic reticulum membrane following translation. Ablation by RNA interference of both TRAPbeta and translocon adaptor SEC61beta was subsequently found to inhibit STING's ability to stimulate expression of IFN-beta. Thus, as well as identifying a regulator of innate immune signalling, our results imply a potential role for the translocon in innate signalling pathways activated by select viruses as well as intracellular DNA.


Resolution of inflammation in inflammatory bowel disease.
Treatment of inflammatory bowel disease at present mainly targets mediators of inflammation to stop or suppress pro-inflammatory processes. Typical examples are steroids, suppression of T cells by thioguanine nucleotides, or antibodies against cytokines such as tumour necrosis factor, interleukin 12, or interleukin 23. In addition to suppression of inflammation, development of therapeutic strategies that support resolution of inflammation or that actively resolve inflammation might be desirable. Resolution of inflammation is now seen as an active process involving specific mediators (eg, lipid mediators or specific cytokines) that is mandatory to restore organ function and completely shut down inflammation. The molecular pathways involved in resolution of inflammation have been investigated in recent years and could be adopted in treatment strategies for inflammatory bowel disease. Among these approaches are anti-integrin strategies and means to produce or locally increase restitution or resolution factors, such as restoration of the activity of transforming growth factor-β by anti-SMAD7 antisense oligonucleotides. The potential role of inflammation-resolving lipid mediators (eg, resolvins), however, still warrants further study and clinical development. This Review focuses on the specific role of active resolution of inflammation in inflammatory bowel disease pathophysiology. Potential therapeutic targets based on these pathways are also discussed.


Far-Red Emitting Fluorescent Dyes for Optical Nanoscopy: Fluorinated Silicon-Rhodamines (SiRF Dyes) and Phosphorylated Oxazines.
Far-red emitting fluorescent dyes for optical microscopy, stimulated emission depletion (STED), and ground-state depletion (GSDIM) super-resolution microscopy are presented. Fluorinated silicon-rhodamines (SiRF dyes) and phosphorylated oxazines have absorption and emission maxima at about λ≈660 and 680 nm, respectively, possess high photostability, and large fluorescence quantum yields in water. A high-yielding synthetic path to introduce three aromatic fluorine atoms and unconventional conjugation/solubilization spacers into the scaffold of a silicon-rhodamine is described. The bathochromic shift in SiRF dyes is achieved without additional fused rings or double bonds. As a result, the molecular size and molecular mass stay quite small (<600 Da). The use of the λ=800 nm STED beam instead of the commonly used one at λ=750-775 nm provides excellent imaging performance and suppresses re-excitation of SiRF and the oxazine dyes. The photophysical properties and immunofluorescence imaging performance of these new far-red emitting dyes (photobleaching, optical resolution, and switch-off behavior) are discussed in detail and compared with those of some well-established fluorophores with similar spectral properties.


The thyroglobulin gene: the third locus for autoimmune thyroid disease or a false dawn?
Genetic studies have identified the HLA and CTLA4 regions as susceptibility loci for the development of common autoimmune thyroid diseases (AITDs), including Graves' disease and autoimmune hypothyroidism. Despite numerous studies, the identification of a third locus has remained elusive. Genetic-linkage studies have implicated chromosome 8q24 as a susceptibility locus for AITD. The gene encoding thyroglobulin (Tg), which encodes a major thyroid autoantigen, maps to this region, and a recent study has reported the association of several exonic single-nucleotide polymorphisms (SNPs) with disease. Although these preliminary data are potentially exciting, caution needs to be exercised, and replication of the data sought before Tg can be designated as the third locus for AITD.


Ultrahigh speed photography of picosecond light pulses and echoes.
Three new results have been obtained with a recently developed camera of 10-psec framing time: (1) The effect of the finite speed of light in photographing relativistic objects is experimentally demonstrated, by photographing a dumbbell-like entity formed by two packets of light. In contrast to material objects, which, theory predicts, should appear rotated, the light dumbbell appears sheared. (2) Photographs of the mode-locked Nd: glass laser radiation show numerous subsidiary pulses accompanying the main ultrashort pulses in the train. The latter have durations ranging from 7 psec to 15 psec. (3) The technique of gated picture ranging, previously used with nanosecond pulses, is extended to the picosecond range where a resolution of 1 cm is demonstrated. Some potentially useful applications are proposed.


The regulation of synaptic strength within motor units of the frog cutaneous pectoris muscle.
The physiological properties of frog neuromuscular junctions may vary widely in a single muscle. In order to understand the factors that contribute to this variation, we have studied populations of synapses belonging to individual motor units of the frog cutaneous pectoris muscle. Motor units in this muscle differ widely in twitch strength. A motor axon's synaptic contacts could be found throughout the muscle, at both singly and polyneuronally innervated endplates. Indeed, over 36% of the endplates contacted by each isolated motor axon were polyneuronally innervated. Comparisons of synapses on muscle fibers in large twitch motor units with those in small twitch motor units reveal that endplate potential amplitude, transmitter release, and muscle fiber diameter are positively correlated with the strength of the motor unit contraction. Large and small twitch motor units differ more in their transmitter release than in their nerve terminal length, indicating that larger twitch motor units have a higher release per unit length of terminal. Among motor units of roughly similar twitch tension, transmitter release at endplates receiving only one axonal input is remarkably constant, independent of postsynaptic muscle fiber input resistance, or, presumably, nerve terminal size. In cases where two different motor axons contribute to a single endplate, the synaptic strength of each input is again related to properties of the contributing motoneuron, although the individual synaptic inputs are markedly reduced in strength and size relative to singly innervated endplates. Additionally, the diameter of polyneuronally innervated muscle fibers appears related to properties of both innervating motoneurons. Thus, the pre- and postsynaptic characteristics of neuromuscular junctions may be determined both by the motoneuron and by peripheral interactions between motoneurons.


Bed bugs evolved unique adaptive strategy to resist pyrethroid insecticides.
Recent advances in genomic and post-genomic technologies have facilitated a genome-wide analysis of the insecticide resistance-associated genes in insects. Through bed bug, Cimex lectularius transcriptome analysis, we identified 14 molecular markers associated with pyrethroid resistance. Our studies revealed that most of the resistance-associated genes functioning in diverse mechanisms are expressed in the epidermal layer of the integument, which could prevent or slow down the toxin from reaching the target sites on nerve cells, where an additional layer of resistance (kdr) is possible. This strategy evolved in bed bugs is based on their unique morphological, physiological and behavioral characteristics and has not been reported in any other insect species. RNA interference-aided knockdown of resistance associated genes showed the relative contribution of each mechanism towards overall resistance development. Understanding the complexity of adaptive strategies employed by bed bugs will help in designing the most effective and sustainable bed bug control methods.


Role of glycolysis in adenylate depletion and repletion during work and recovery in teleost white muscle.
Measurements of metabolite concentrations before and immediately after swimming of trout to exhaustion indicate that all three potential endogenous fuels of anaerobic metabolism [glycogen, phosphocreatine (PCr) and adenosine triphosphate (ATP)] are utilized during anaerobic white muscle work. Lactate, H+, creatine Pi, NH4+ and inosine monophosphate (IMP) are formed in the process. Glycolysis is considered to be functionally (if loosely) coupled to adenylate depletion by setting up conditions favouring AMP-deaminase-catalysed formation of IMP and NH3. During recovery under these experimental conditions, glycolysis appears to outcompete oxidative metabolism as an ADP acceptor; therefore, in this kind of white muscle, glycolysis is also linked to IMP reconversion to AMP and thus to adenylate replenishment. The net process generates H+, which is why ATP replenishment must be completed before PCr concentrations can be returned to pre-exercise values.


The Neuroprotective Role of Origanum syriacum L. and Lavandula dentata L. Essential Oils through Their Effects on AMPA Receptors.
Lavandula dentata L. and Origanum syriacum L. essential oils have numerous health benefits and properties, such as possessing common components with a variant degree of depressive actions in the central nervous system. We investigated the depressive property of these oils on AMPA receptors, which are responsible for most of the fast-excitatory neurotransmission in the CNS and play a critical role in synaptic plasticity. Since excessive activation of AMPARs has been linked to neurotoxicity leading to various pathologies, we hypothesize that these oils have a neuroprotective role by acting directly on the kinetics of AMPARs. Using Gas Chromatography-Mass Spectrometry (GC/MS) and patch-clamp electrophysiology, the essential oils of L. dentata flowers and O. syriacum leaves were characterized and the whole cell currents were measured with and without the administration of the oils onto HEK293 cells. The current study results showed that the biophysical properties of AMPA receptor subunits showed a decrease in desensitization rate of GluA1 and GluA2 homomers, using O. syriacum, while administering L. dentata oil decreased the desensitization rate of GluA1 and GluA2 homomers, as well as GluA1/2 heteromers. As for the deactivation rate, both oils slowed the deactivation kinetics of all AMPA receptor subunits. Intriguingly, between the two oils, the effect of desensitization and deactivation was of a greater significance for L. dentata oil than O. syriacum. Our data suggest that the two oils contain components that are essential to identify, as those active components underlie the oils' neuronal depressive properties reported, and to extract them to synthesize a potent neuroprotective drug to treat neurological diseases potentially.


Transcutaneous Vagus Nerve Stimulation Modulates Default Mode Network in Major Depressive Disorder.
Depression is the most common form of mental disorder in community and health care settings and current treatments are far from satisfactory. Vagus nerve stimulation (VNS) is a Food and Drug Administration approved somatic treatment for treatment-resistant depression. However, the involvement of surgery has limited VNS only to patients who have failed to respond to multiple treatment options. Transcutaneous VNS (tVNS) is a relatively new, noninvasive VNS method based on the rationale that there is afferent/efferent vagus nerve distribution on the surface of the ear. The safe and low-cost characteristics of tVNS have the potential to significantly expand the clinical application of VNS. In this study, we investigated how tVNS can modulate the default mode network (DMN) functional connectivity (FC) in mild or moderate major depressive disorder (MDD) patients. Forty-nine MDD patients were recruited and received tVNS or sham tVNS (stVNS) treatments. Thirty-four patients completed the study and were included in data analysis. After 1 month of tVNS treatment, the 24-item Hamilton Depression Rating Scale score reduced significantly in the tVNS group as compared with the stVNS group. The FC between the DMN and anterior insula and parahippocampus decreased; the FC between the DMN and precuneus and orbital prefrontal cortex increased compared with stVNS. All these FC increases are also associated with 24-item Hamilton Depression Rating Scale reduction. tVNS can significantly modulate the DMN FC of MDD patients; our results provide insights to elucidate the brain mechanism of tVNS treatment for MDD patients.


Adequacy of energy intake among breast-fed infants in the DARLING study: relationships to growth velocity, morbidity, and activity levels. Davis Area Research on Lactation, Infant Nutrition and Growth.
Breast-fed infants grow less rapidly after the first 2 to 3 months of age than current standards. The DARLING study (Davis Area Research on Lactation, Infant Nutrition and Growth) was designed to evaluate whether this pattern should be considered "faltering" or is a normal outcome even under optimal conditions. Data on intake, growth, morbidity, activity, and motor development were collected longitudinally from infants who were breast fed for at least 12 months. Gross energy intake, calculated from 4-day records of milk and food intake at 3, 6, 9, and 12 months, averaged 91.4, 84.1, 86.7, and 91.8 kcal/kg per day, respectively, well below recommended amounts of metabolizable energy. Nevertheless, infants usually left some food unconsumed. Growth velocity was also below current reference data and was weakly correlated with energy intake. There were no significant negative associations between energy intake at any time and incidence, prevalence, or duration of any category of morbidity during the subsequent 3 months. There were no consistent associations between energy intake and activity level, time spent sleeping, or achievement of key developmental milestones. Similarly, infants with slower growth velocity were just as active and were ill no more often in subsequent months than infants who were growing more rapidly. Thus the deviation from current recommendations for energy intake and growth can be considered a normal pattern with no apparent deleterious consequences in our population of breast-fed infants.


Blood transcriptomics to characterize key biological pathways and identify biomarkers for predicting mortality in melioidosis.
Melioidosis is an often lethal tropical disease caused by the Gram-negative bacillus, Burkholderia pseudomallei. The study objective was to characterize transcriptomes in melioidosis patients and identify genes associated with outcome. Whole blood RNA-seq was performed in a discovery set of 29 melioidosis patients and 3 healthy controls. Transcriptomic profiles of patients who did not survive to 28 days were compared with patients who survived and healthy controls, showing 65 genes were significantly up-regulated and 218 were down-regulated in non-survivors compared to survivors. Up-regulated genes were involved in myeloid leukocyte activation, Toll-like receptor cascades and reactive oxygen species metabolic processes. Down-regulated genes were hematopoietic cell lineage, adaptive immune system and lymphocyte activation pathways. RT-qPCR was performed for 28 genes in a validation set of 60 melioidosis patients and 20 healthy controls, confirming differential expression. IL1R2, GAS7, S100A9, IRAK3, and NFKBIA were significantly higher in non-survivors compared with survivors (P < 0.005) and healthy controls (P < 0.0001). The AUROCC of these genes for mortality discrimination ranged from 0.80-0.88. In survivors, expression of IL1R2, S100A9 and IRAK3 genes decreased significantly over 28 days (P < 0.05). These findings augment our understanding of this severe infection, showing expression levels of specific genes are potential biomarkers to predict melioidosis outcomes.


A Four-kallikrein Panel Predicts High-grade Cancer on Biopsy: Independent Validation in a Community Cohort.
A statistical model based on four kallikrein markers (total prostate-specific antigen [tPSA], free PSA [fPSA], intact PSA, and human kallikrein-related peptidase 2) in blood can predict risk of Gleason score ≥7 (high-grade) cancer at prostate biopsy. To determine the value of this model in predicting high-grade cancer at biopsy in a community-based setting in which referral criteria included percentage of fPSA to tPSA (%fPSA). We evaluated the model, with or without adding blood levels of microseminoprotein-β (MSMB) in a cohort of 749 men referred for prostate biopsy due to elevated PSA (≥3 ng/ml), low %fPSA (<20%), or suspicious digital rectal examination at Skåne University Hospital, Malmö, Sweden. The kallikrein markers, with or without MSMB levels, measured in cryopreserved anticoagulated blood were combined with age in a published statistical model (Prostate Testing for Cancer and Treatment [ProtecT]) to predict high-grade cancer at biopsy. Predictive accuracy was compared with a base model. The %fPSA was low (median: 17; interquartile range: 13-22) in this cohort because this marker was used as a referral criterion. The ProtecT model improved discrimination over age and PSA for high-grade cancer (0.777 vs 0.720; p=0.002). At one illustrative cut point, use of the panel would reduce the number of biopsies by 236 per 1000 and detect 195 of 208 (94%) but delay diagnosis of 13 of 208 high-grade cancers. MSMB levels in blood did not improve the accuracy of the panel (p=0.2). The kallikrein model is predictive of high-grade cancer if criteria for biopsy referral also include %fPSA, and it can reduce unnecessary biopsies without missing an undue number of tumors. We evaluated a published model to predict biopsy outcome in men biopsied due to low percentage of free to total prostate-specific antigen. The model helps reduce unnecessary biopsies without missing an undue number of high-grade cancers.


[BODY COMPOSITION AND SOMATOTYPE IN UNIVERSITY TRIATHLETES].
the triathlon is an endurance sport and individual that consists of three different disciplines: swimming, cycling and running. The aim of the study was to describe and analyze the anthropometric characteristics, body composition and somatotype in male college triathletes. observational and descriptive study of anthropometric characteristics, body composition and somatotype of 39 male college athletes from 24 ± 4,5 years, participants in the championship of Spain university triathlon sprint mode (Alicante 2010), from different universities Spanish. According to anthropometric measurement techniques adopted by the International Society for the Advancement of Kinanthropometry (ISAK) and the Spanish Group Cineantropometría (GREC) by an accredited assessor ISAK Level II. we find athletes of stunting, where you destacanvalores below normal in the subscapularis, supraspinatus, triceps and biceps skinfold, percentage of muscle mass (45.27 ± 3.29%) and fat mass (10.22 ± 2.92%) and bone (16.65 ± 1.34%) and where mesomorphy somatotipo predominates. the triathletes and runners have lower size that cyclists and swimmers. Triathletes and cyclists show a similar weight, less than swimmers line, and more than 10km runners. Iliac crest skinfold, abdominal and thigh front cyclists are less than triathletes. The percentage of fat mass of runners triathletes and swimmers are similar, however the muscle mass of athletes usually less than cyclists but similar to other forms. Somatotype resembles triathlete cyclist (mesomorph). The corridor is ectomorph and mesomorph-swimmer can range from a ectomorph mesomorph.


Characterization of metabolites of xylazine produced in vivo and in vitro by LC/MS/MS and by GC/MS.
The metabolic fate of xylazine, 2-(2,6-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazine, in horses is described. The major metabolites identified in the hydrolyzed horse urine were 2-(4'-hydroxy-2',6'-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazi ne, 2-(3'-hydroxy-2',6'-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazi ne, N-(2,6-dimethylphenyl)thiourea, and 2-(2',6'-dimethylphenylamino)-4-oxo-5,6-dihydro-1,3-thiazine. These metabolites were also produced by incubating xylazine with rat liver microsomes. The major metabolite produced in vitro by rat liver preparations was found to be the ring opened N-(2,6-dimethylphenyl)thiourea. The identities of these metabolites were confirmed by spectroscopic comparisons with synthetic standards. Phenolic metabolic standards were synthesized efficiently by the use of Fenton's reagent. This reagent was used to monohydroxylate multiply substituted aromatic ring systems. LC/MS/MS, with an atmospheric pressure chemical ionization source, was found to be particularly useful in confirming the presence of phenolic metabolites in hydrolyzed equine urine and microsomal extracts. These phenolic metabolites could not be analyzed by GC/MS even after derivatization with silylating agents. The advantage of LC/MS/MS was that no or little sample preparation of urine or microsomal extract was necessary prior to the analysis. A mechanism is also proposed for the formation of the major metabolite, N-(2,6-dimethylphenyl)thiourea, from xylazine.


The influence of different implant materials on human gingival fibroblast morphology, proliferation, and gene expression.
The aim of this study was to investigate the cellular response of human gingival fibroblasts (HGFs) cultured on smooth or rough zirconia (Zr) or titanium (Ti) disks. Disks fabricated from four different materials--smooth Zr (Zr-S), rough Zr (Zr-R), smooth Ti (Ti-S), and rough Ti (Ti-R)--were used, and surface roughness was analyzed by atomic force microscopy. After HGFs were cultured on these disks, cell morphology was examined by scanning electron microscopy, cell proliferation activity was evaluated by a monotetrazolium assay, and gene expression levels of various collagens and integrins were measured by real-time polymerase chain reaction. The Ti-R disks were the roughest, followed by Zr-R, Ti-S, and Zr-S, in that order. The cells cultured on the Zr-S and Ti-S disks appeared to be more aligned with the fine irregularities at later time points, whereas the cells cultured on the Zr-S showed the weakest spreading compared to the other surfaces after 3 hours of culture. With respect to proliferation, cells proliferated significantly faster on the Zr-S surface than on the other surfaces. Gene expression of integrin α2 at 3 hours and integrin α5 and type I collagen at 48 hours on Zr-S was significantly up-regulated compared to Ti. Conversely, the expression of integrins β1 and β3 and type III collagen was significantly decreased on Zr-S at 1 hour compared to the other materials. These data indicate that different surface materials and topographies may induce a distinct HGF morphology, proliferation, and gene expression.


A topological interaction between cohesin rings and a circular minichromosome.
Sister chromatid cohesion depends on a multiprotein cohesin complex containing two SMC subunits, Smc1 and Smc3, that dimerize to form V-shaped molecules with ABC-like ATPase heads at the tips of their two arms. Cohesin's Smc1 and Smc3 "heads" are connected by an alpha kleisin subunit called Scc1, forming a tripartite ring with a diameter around 40 nm. We show here that some cohesin remains tightly bound to circular minichromosomes after their purification from yeast cells and that cleavage either of cohesin's ring or of the minichromosome's DNA destroys their association. This suggests that the stable association between cohesin and chromatin detected here is topological rather than physical, which is consistent with the notion that DNA is trapped inside cohesin rings.


Significant variations in nuclear structure occur between and within Gleason grading patterns 3, 4, and 5 determined by digital image analysis.
Alterations in nuclei structure and DNA content captured from Gleason grading patterns 3, 4 and 5 of radical prostatectomy (RP) cases were determined by a computer-assisted microscope. Quantitative Nuclear Morphometry (QNM) profiles were created to evaluate variability in nuclear structure within each of these grades. A tissue microarray (TMA) was constructed using RP cases and the prostate cancer (PCa) TMA cores prepared from 20 GG-3, 9 GG-4, 10 GG-5 patterns, and 20 benign cancer-adjacent cases from RP archival paraffin blocks. Feulgen-stained nuclei were captured from 0.6 mm spots using the AutoCyte system. Pools of 1100 nuclei captured from each test group were used to calculate Multivariate Logistic Regression (MLR) models that generated predictive indices and predictive probabilities (PP) to make comparisons between and within each set of pooled nuclei. A single QNM profiles yielded areas of receiver operator characteristic curves (ROC) that distinguished differences among benign cancer-adjacent nuclei and GG-3 (ROC-AUC = 0.78); GG-4 (ROC-AUC = 0.86) and GG-5 (ROC-AUC = 0.88) with accuracies of 73%, 78% and 80% respectively. Applying PP plots generated from MLR models of GG 3, 4, and 5 nuclei clearly demonstrated marked heterogeneity within each of these three GG patterns. QNM signatures illustrate alterations in nuclei structure, based upon nuclear morphometry within each of these three GG patterns, and might signify potential variations in PCa disease risk of progression outcomes. In the future a modified system of Gleason grading that considers not only glandular architecture but also quantitative nuclear grade may ensure accuracy in prognosis.


Novel Deleterious Dihydropyrimidine Dehydrogenase Variants May Contribute to 5-Fluorouracil Sensitivity in an East African Population.
Clinical studies have identified specific genetic variants in dihydropyrimidine dehydrogenase (DPD; DPYD gene) as predictors of severe adverse toxicity to the commonly used chemotherapeutic 5-fluorouracil (5-FU); however, these studies have focused on European and European-American populations. Our laboratory recently demonstrated that additional variants in non-European haplotypes are predictive of 5-FU toxicity. The objective of this study was to identify potential risk variants in an understudied East African population relevant to our institution's catchment area. The DPYD protein-coding region was sequenced in 588 individuals of Somali or Kenyan ancestry living in central/southeast Minnesota. Twelve novel nonsynonymous variants were identified, seven of which significantly decreased DPD activity in vitro. The commonly reported toxicity-associated variants, *2A, D949V, and I560S, were not detected in any individuals. Overall, this study demonstrates a critical limitation in our knowledge of pharmacogenetic predictors of 5-FU toxicity, which has been based on clinical studies conducted in populations of limited diversity.


Opioid pharmacogenomics using a twin study paradigm: methods and procedures for determining familial aggregation and heritability.
Opioids are the cornerstone medication for the treatment of moderate to severe pain. However, analgesic opioid requirements and the propensity to suffer from aversive opioid effects, including fatal respiratory depression and addiction, vary widely among patients. The factors underlying the substantial response variance remain largely unknown and need clarification for using opioids more effectively in appropriately selected patients. This ongoing study takes advantage of the twin paradigm to estimate the genetic and environmental contributions to inter-individual differences in opioid responses. Evidence of significant heritability will justify more detailed and extensive genomic studies. The enrollment target is 80 monozygotic and 45 dizygotic twin pairs who undergo a target-controlled infusion of the opioid alfentanil and saline placebo in sequential but randomized order. In a laboratory-type setting, well-defined pharmacodynamic endpoints are measured to quantify pain sensitivity, analgesic opioid effects, and aversive opioid effects including respiratory depression, sedation and reinforcing affective responses. First results obtained in 159 participants provide evidence for the feasibility and utility of this interventional study paradigm to estimate familial aggregation and heritability components of relevant drug effects. Areas highlighted in this report include recruitment strategies, required infrastructure and personnel, selection of relevant outcome measures, drug infusion algorithm minimizing pharmacokinetic variability, and considerations for optimizing data quality and quantity without hampering feasibility. Applying the twin paradigm to complex and potentially harmful studies comprehensively characterizing pharmacological response profiles is without much precedent. Methods and first results including heritability estimates for heat and cold pain sensitivity should be of interest to investigators considering similar studies.


A critical evaluation of the validity and the reliability of global competency constructs for supervisor assessment of junior medical trainees.
Supervisor assessments are critical for both formative and summative assessment in the workplace. Supervisor ratings remain an important source of such assessment in many educational jurisdictions even though there is ambiguity about their validity and reliability. The aims of this evaluation is to explore the: (1) construct validity of ward-based supervisor competency assessments; (2) reliability of supervisors for observing any overarching domain constructs identified (factors); (3) stability of factors across subgroups of contexts, supervisors and trainees; and (4) position of the observations compared to the established literature. Evaluated assessments were all those used to judge intern (trainee) suitability to become an unconditionally registered medical practitioner in the Australian Capital Territory, Australia in 2007-2008. Initial construct identification is by traditional exploratory factor analysis (EFA) using Principal component analysis with Varimax rotation. Factor stability is explored by EFA of subgroups by different contexts such as hospital type, and different types of supervisors and trainees. The unit of analysis is each assessment, and includes all available assessments without aggregation of any scores to obtain the factors. Reliability of identified constructs is by variance components analysis of the summed trainee scores for each factor and the number of assessments needed to provide an acceptably reliable assessment using the construct, the reliability unit of analysis being the score for each factor for every assessment. For the 374 assessments from 74 trainees and 73 supervisors, the EFA resulted in 3 factors identified from the scree plot, accounting for only 68 % of the variance with factor 1 having features of a "general professional job performance" competency (eigenvalue 7.630; variance 54.5 %); factor 2 "clinical skills" (eigenvalue 1.036; variance 7.4 %); and factor 3 "professional and personal" competency (eigenvalue 0.867; variance 6.2 %). The percent trainee score variance for the summed competency item scores for factors 1, 2 and 3 were 40.4, 27.4 and 22.9 % respectively. The number of assessments needed to give a reliability coefficient of 0.80 was 6, 11 and 13 respectively. The factor structure remained stable for subgroups of female trainees, Australian graduate trainees, the central hospital, surgeons, staff specialist, visiting medical officers and the separation into single years. Physicians as supervisors, male trainees, and male supervisors all had a different grouping of items within 3 factors which all had competency items that collapsed into the predefined "face value" constructs of competence. These observations add new insights compared to the established literature. For the setting, most supervisors appear to be assessing a dominant construct domain which is similar to a general professional job performance competency. This global construct consists of individual competency items that supervisors spontaneously align and has acceptable assessment reliability. However, factor structure instability between different populations of supervisors and trainees means that subpopulations of trainees may be assessed differently and that some subpopulations of supervisors are assessing the same trainees with different constructs than other supervisors. The lack of competency criterion standardisation of supervisors' assessments brings into question the validity of this assessment method as currently used.


Adaptive frequency distribution for electrical bioimpedance spectroscopy measurements.
This paper presents a novel frequency distribution scheme intended to provide more accurate estimations of Cole parameters. Nowadays a logarithmic frequency distribution is mostly used in Electrical Bioimpedance Spectroscopy (EBIS) applications. However it is not optimized following any criterion. Our hypothesis is that an EBIS signal contains more information where the variation of the measurement regarding the frequency is larger; and that there ought to be more measuring frequencies where there is more information. Results show that for EBIS data with characteristic frequencies up to 200 kHz the error obtained with both frequency distribution schemes is similar. However, for EBIS data with higher values of characteristic frequency the error produced when estimating the values from EBIS measurements using an adaptive frequency distribution is smaller. Thus it may useful for EBIS applications with high values of characteristic frequency, e.g. cerebral bioimpedance.


Personal Trust and System Trust in the Sharing Economy: A Comparison of Community- and Platform-Based Models.
Currently, new business models created in the sharing economy differ considerably and they differ in the formation of trust as well. If and how trust can be created is shown by a comparison of two examples which diverge in their founding philosophy. The chosen example of community-based economy, Community Supported Agriculture (CSA), no longer trusts the capitalist system and therefore distances itself and creates its own environment including a new business model. It is implemented within rather small groups where trust is created by personal relations and face-to-face communication. On the contrary, the example of a platform economy, the accommodation-provider company Airbnb, shows trust in the system and pushes technological innovations through the use of platform applications. It promotes trust and confidence in the progress of technology. For the conceptual analysis, the distinction between personal trust and system trust defined by Niklas Luhmann is adopted. The analysis describes two different modes of trust formation and how they push distrust or improve trust. Grounded on these analyses, assumptions on the process of trust formation within varying models of the sharing economy are formulated as well as a hypothesis about possible developments is introduced for further research.


Development of the rat telencephalon--volumetric analysis.
With regard to intensive morphometric changes, morphometry as a method is mainly used for histogenetic studies of brain development in normal and experimental conditions. The aim of our study was to quantitatively analyse morphological parameters of the rat telencephalon during embryonic development. The investigation was carried out on semithin serial sections of rat brain from embryonic days 12 to 15. The volume densities (VV) of the lateral ventricles, the telencephalic neuroepithelium and the surrounding mesenchyme have been analysed stereologically and compared in examined embryonic stages. The neuroepithelial volume density was the smallest (28%) at E13 and the biggest (44%) at E15 (p<0.0005). The mesenchymal volume density was the smallest (32%) at E13 and the biggest (48%) at E14 (p<0.0005). The volume density of lateral ventricles was the biggest (40%) at E13 and the smallest (14%) at E15 (p<0.0005). Neurostereological methods have been making a very valuable contribution to neuroscience over recent years. We have used unbiased stereological counting methods to obtain objective quantitative parameters which show relations between some parts of rat embryonic telencephalon examined during its normal development.


Bilateral Septic Neonatal Orchiepididymitis - Case Report.
Neonatal epididymo-orchitis is a rare condition, causing testicular pain in neonatal boys. It represents epididymo-testicular inflammation which commonly coexists with urinary tract infections and malformations. The idiopathic type is extremely rare. We present a case of a seven-day old male neonate with advanced septic form of idiopathic orchiepididymitis and no associated urinary tract anomalies. The boy was hospitalized with signs of sepsis, anterior abdominal wall phlegmona and bilateral acute scrotum. Colour Doppler echosonography indicated epididymo-testicular inflammation with increased vascular flow. The patient underwent surgical exploration of both scrota in order to evacuate purulent content and fibrin. Cultures of Enterobacter spp were detected in hemiscrotal pus. Prompt administration of antibiotics was done. The postoperative course was uneventful. We suggest that every male baby must be very meticulously examined by a neonatologist in the early postnatal period, in order to prevent infertility.


Treatment of median arcuate ligament syndrome: outcome of laparoscopic approach.
Median arcuate ligament syndrome(MALS) is a rare condition thatmay cause significant clinical manifestations, including abdominal pain and weight loss. Its diagnosis may be difficult and very often delayed. The laparoscopic approach became the standard treatment of MALS. To assess the outcome of laparoscopic treatment in patients with MALS. The data of sixpatients with MALS who were subjected to laparoscopic sectioning of the median arcuate ligament were retrospectively reviewed.The following data were evaluated: age, gender, clinical and diagnostic tests findings, ASA score, operative findings and complications, postoperative complications and mortality, hospital stay duration, and hospital readmission.The diagnosis of MALS was established by CT angiography and/or MR angiography. There were four (66.7%) women and two (33.3%) men aged from 32 to 60 years. The main symptoms were epigastric pain (100%) and weight loss (66.7%). The findings of high-grade stenosis of the proximal celiac axis and poststenotic dilation confirmed on angiography confirmed the diagnosis in all patients. Surgical procedure was uneventful in all patients. The only postoperative complication was urinary retention that occurred in a male. At three-month follow-up, all patients were asymptomatic. Laparoscopic treatment of MALS is safe and effective in relieving the clinical manifestations of patients.


Suicide rates amongst individuals from ethnic minority backgrounds: A systematic review and meta-analysis.
Existing evidence suggests that some individuals from ethnic minority backgrounds are at increased risk of suicide compared to their majority ethnic counterparts, whereas others are at decreased risk. We aimed to estimate the absolute and relative risk of suicide in individuals from ethnic minority backgrounds globally. Databases (Medline, Embase, and PsycInfo) were searched for epidemiological studies between 01/01/2000 and 3/07/2020, which provided data on absolute and relative rates of suicide amongst ethnic minority groups. Studies reporting on clinical or specific populations were excluded. Pairs of reviewers independently screened titles, abstracts, and full texts. We used random effects meta-analysis to estimate overall, sex, location, migrant status, and ancestral origin, stratified pooled estimates for absolute and rate ratios. PROSPERO registration: CRD42020197940. A total of 128 studies were included with 6,026,103 suicide deaths in individuals from an ethnic minority background across 31 countries. Using data from 42 moderate-high quality studies, we estimated a pooled suicide rate of 12·1 per 100,000 (95% CIs 8·4-17·6) in people from ethnic minority backgrounds with a broad range of estimates (1·2-139·7 per 100,000). There was weak statistical evidence from 51 moderate-high quality studies that individuals from ethnic minority groups were more likely to die by suicide (RR 1·3 95% CIs 0·9-1·7) with again a broad range amongst studies (RR 0·2-18·5). In our sub-group analysis we only found evidence of elevated risk for indigenous populations (RR: 2·8 95% CIs 1·9-4·0; pooled rate: 23·2 per 100,000 95% CIs 14·7-36·6). There was very substantial heterogeneity (I2 > 98%) between studies for all pooled estimates. The homogeneous grouping of individuals from ethnic minority backgrounds is inappropriate. To support suicide prevention in marginalised groups, further exploration of important contextual differences in risk is required. It is possible that some ethnic minority groups (for example those from indigenous backgrounds) have higher rates of suicide than majority populations. No specific funding was provided to conduct this research. DK is funded by Wellcome Trust and Elizabeth Blackwell Institute Bristol. Matthew Spittal is a recipient of an Australian Research Council Future Fellowship (project number FT180100075) funded by the Australian Government. Rebecca Musgrove is funded by the NIHR Greater Manchester Patient Safety Translational Research Centre (PSTRC-2016-003).


Nmyc plays an essential role during lung development as a dosage-sensitive regulator of progenitor cell proliferation and differentiation.
Understanding how lung progenitor cells balance proliferation against differentiation is relevant to clinical disorders such as bronchopulmonary dysplasia of premature babies and lung cancer. Previous studies have established that lung development is severely disrupted in mouse mutants with reduced levels of the proto-oncogene Nmyc, but the precise mechanisms involved have not been explored. We show here that Nmyc expression in the embryonic lung is normally restricted to a distal population of undifferentiated epithelial cells, a high proportion of which are in the S phase of the cell cycle. Overexpression of NmycEGFP in the epithelium under the control of surfactant protein C (Sftpc) regulatory elements expands the domain of S phase cells and upregulates numerous genes associated with growth and metabolism, as shown by transcriptional microarray. In addition, there is marked inhibition of differentiation, coupled with an expanded domain of expression of Sox9 protein, which is also normally restricted to the distal epithelial compartment. By contrast, conditional deletion of Nmyc leads to reduced proliferation, epithelial differentiation and high levels of apoptosis in both epithelium and mesenchyme. Unexpectedly, about 50% of embryos in which only one copy of Nmyc is deleted die perinatally, with similarly abnormal lungs. We propose a model in which Nmyc is essential in the developing lung for maintaining a distal population of undifferentiated, proliferating progenitor cells.


Structure and function of human DnaJ homologue subfamily a member 1 (DNAJA1) and its relationship to pancreatic cancer.
Pancreatic cancer has a dismal 5 year survival rate of 5.5% that has not been improved over the past 25 years despite an enormous amount of effort. Thus, there is an urgent need to identify truly novel yet druggable protein targets for drug discovery. The human protein DnaJ homologue subfamily A member 1 (DNAJA1) was previously shown to be downregulated 5-fold in pancreatic cancer cells and has been targeted as a biomarker for pancreatic cancer, but little is known about the specific biological function for DNAJA1 or the other members of the DnaJ family encoded in the human genome. Our results suggest the overexpression of DNAJA1 suppresses the stress response capabilities of the oncogenic transcription factor, c-Jun, and results in the diminution of cell survival. DNAJA1 likely activates a DnaK protein by forming a complex that suppresses the JNK pathway, the hyperphosphorylation of c-Jun, and the anti-apoptosis state found in pancreatic cancer cells. A high-quality nuclear magnetic resonance solution structure of the J-domain of DNAJA1 combined with a bioinformatics analysis and a ligand affinity screen identifies a potential DnaK binding site, which is also predicted to overlap with an inhibitory binding site, suggesting DNAJA1 activity is highly regulated.


The gene expression analysis of blood reveals S100A11 and AQP9 as potential biomarkers of infective endocarditis.
The diagnostic and prognostic assessments of infective endocarditis (IE) are challenging. To investigate the host response during IE and to identify potential biomarkers, we determined the circulating gene expression profile using whole genome microarray analysis. A transcriptomic case-control study was performed on blood samples from patients with native valve IE (n = 39), excluded IE after an initial suspicion (n = 10) at patient's admission, and age-matched healthy controls (n = 10). Whole genome microarray analysis showed that patients with IE exhibited a specific transcriptional program with a predominance of gene categories associated with cell activation as well as innate immune and inflammatory responses. Quantitative real-time RT-PCR performed on a selection of highly modulated genes showed that the expression of the gene encoding S100 calcium binding protein A11 (S100A11) was significantly increased in patients with IE in comparison with controls (P<0.001) and patients with excluded IE (P<0.05). Interestingly, the upregulated expression of the S100A11 gene was more pronounced in staphylococcal IE than in streptococcal IE (P<0.01). These results were confirmed by serum concentrations of the S100A11 protein. Finally, we showed that in patients with IE, the upregulation of the aquaporin-9 gene (AQP9) was significantly associated with the occurrence of acute heart failure (P = 0.02). Using transcriptional signatures of blood samples, we identified S100A11 as a potential diagnostic marker of IE, and AQP9 as a potential prognostic factor.


Microfluidic on-demand engineering of exosomes towards cancer immunotherapy.
Extracellular vesicles (EVs), particularly exosomes (30-150 nm), are an emerging delivery system in mediating cellular communications, which have been observed for priming immune responses by presenting parent cell signaling proteins or tumor antigens to immune cells. Therefore, preparation of antigenic exosomes that can play therapeutic roles, particularly in cancer immunotherapy, is emerging. However, standard benchtop methods (e.g., ultracentrifugation and filtration) lack the ability to purify antigenic exosomes specifically among other microvesicle subtypes, due to the non-selective and time-consuming (>10 h) isolation protocols. Exosome engineering approaches, such as the transfection of parent cells, also suffer from poor yields, low purity, and time-consuming operations. In this paper, we introduce a streamlined microfluidic cell culture platform for integration of harvesting, antigenic modification, and photo-release of surface engineered exosomes in one workflow, which enables the production of intact, MHC peptide surface engineered exosomes for cytolysis activation. A PDMS microfluidic cell culture chip is simply cast from a 3D-printed mold. This proof-of-concept study demonstrated the enhanced ability of harvested exosomes in antigen presentation and T cell activation, by decorating melanoma tumor peptides on the exosome surface (e.g., gp-100, MART-1, and MAGE-A3). Such surface engineered antigenic exosomes were harvested in real-time from the on-chip culture of leukocytes isolated from human blood, leading to much faster cellular uptake. The activation of gp100-specific CD8 T cells which were purified from the spleen of 2 Pmel1 transgenic mice was evaluated using surface engineered exosomes prepared from murine antigen presenting cells. Antigen-specific CD8 T cell proliferation was significantly induced by the engineered exosomes compared to that by native, non-engineered exosomes. This microfluidic platform serves as an automated and highly integrated cell culture device for rapid and real-time production of therapeutic exosomes that could advance cancer immunotherapy.


Strength of Plantar- and Dorsiflexors Mediates Step Regularity During a High Cognitive Load Situation in a Cross-sectional Cohort of Older and Younger Adults.
Completing simultaneous tasks while standing or walking (ie, a high cognitive load situation [HCLS]) is inevitable in daily activities and can lead to interference in task performances. Age-related physical and cognitive changes may confound performance variability during HCLS in older and younger adults. Identification of these confounding effects may reveal therapy targets to maintain optimal physical function later in life. The aim of this study was to investigate the effect of increasing the difficulty levels of an additional motor task and restricting visual information, on gait parameters in younger and older adults while considering the effect of cognitive and physical covariates. Fifteen healthy younger and 14 healthy older adults were asked to complete assessments of cognitive function, balance, and strength. They were then asked to walk on a self-paced treadmill with or without carrying a plastic tray. Opaqueness of the tray (vision) and the presence of water in glasses placed on the tray (increasing task difficulty) were varied. Mean, standard deviation, and regularity (sample entropy) of step width and length were compared across conditions and groups using repeated-measures analyses of variance with and without covariate analysis. Only significantly correlated covariates of cognition, balance, and strength were entered into each model. Older adults had greater step width irregularity compared with younger adults across all conditions when controlling for concentric plantar- and dorsiflexion strength. A decline in strength may likely alter neuromuscular control of gait, specifically control of step width, which has been associated with fall risk in older adults. Adjusting for the same covariates revealed increased regularity of step length, as visual feedback from the feet was restricted. Specifically, step length was more regular while carrying an opaque tray compared with not carrying a tray. Visual restriction was a contributing factor, which led to more predictable gait kinematics, indicating the role of sensory information to enhance the adaptability during walking under HCLS. The knowledge of the regularity behavior of human movement can expand physical therapists' treatment approaches to promote further interactivity and coordination across body systems that model behavior of healthy young individuals. Targeting strength during therapy may provide additional benefits for gait performance under HCLS.


Circumferential stretching of saphenous vein smooth muscle enhances vasoconstrictor responses by Rho kinase-dependent pathways.
Surgical preparation and/or pulsatile arterial perfusion of saphenous vein increases the sensitivity of vein rings to calcium mobilising agonists such as phenylephrine. We have investigated the mechanism(s) underlying this effect. We have used an ex vivo flow circuit, with simulated arterial or venous flows (mean pressure 100 and 20 mmHg, respectively), to investigate the sensitivity of human saphenous vein to phenylephrine, 5-hydroxytryptamine (5-HT) and KCl, using organ chamber pharmacology. After 90 min of pulsatile arterial perfusion the mean maximum tension induced by KCl had increased from 4.7 to 11.1 g (n=5), by phenylephrine had increased from 4.4 to 10.2 g (n=8) and by 5-HT had increased from 4.4 to 6.7 g (n=10), all P<0.01. Phenylephrine did not augment the tension in vein rings maximally precontracted with KCl (n=4). The EC(50) for KCl was unchanged after pulsatile arterial perfusion (n=5), but for phenylephrine and 5-HT there were significant reductions from 14+/-5 to 2+/-1 microM (n=8) and from 1.0+/-0.4 to 0.20+/-0.06 microM (n=10), respectively. The rate of contraction (in response to 3 microM phenylephrine) increased from 0.11 g/min to 0.37 g/min, P<0.02, after arterial perfusion (n=4). These changes in contractile properties (to phenylephrine) were endothelium-independent, evident within 5 min of simulated arterial perfusion. The changes in contractile properties could be abrogated by external stenting of the vein (to attenuate circumferential deformation) or inclusion in the perfusate of a vasodilator, e.g., cromakalim (5 microM) or the selective Rho kinase inhibitor Y-27632 (20 microM). The heightened sensitivity and contractility to phenylephrine was maintained after inclusion of adenosine (100 microM), gadolinium (10 microM) or cycloheximide (10 microM) in the vein perfusate. The circumferential deformations imposed by simulated arterial perfusion alter the vasomotor responses of saphenous vein smooth muscle. These effects are independent of new protein synthesis or the activation of stretch activated cation channels. The Rho kinase pathway appears to mediate the signalling mechanisms leading to increased agonist-induced tension and the increased sensitivity to vasoconstrictors.


Controlled Nanopores in Thin Films of Nonstoichiometrically Supramolecularly Assembled Graft Copolymers.
Herein, nanometer-scale morphologies of graft-copolymer-like supramolecular thin films, composed of sulfonic acid terminated polystyrene (SPS) and poly(2-vinylpyridine) (P2VP), and their application to antireflection coatings were investigated. The intermolecular complexes of SPS and P2VP, formed through nonstoichiometric multiple hydrogen bonding between the sulfonic acid group of SPS and the nitrogen atom in pyridine unit of P2VP, occurring in film deposition allowed for the formation of spherical micelles (with SPS and P2VP as the corona and core, respectively) in the thin film. Interestingly, the domain size of the micelles was tunable from approximately 20 to 90 nm on average by controlling either the blend ratio of components or the concentration of polymer solution. Furthermore, nanoporous thin films could be easily prepared by removing the core of micelle-based nanostructures by using a simple solvent etching process, leaving sulfonic acid groups on the surface of nanopores, which can be utilized as potential functional sites. Those resultant nanoporous thin films were conveniently employed as an antireflection layer on a glass substrate, giving a maximum 97.8 % transmittance in the visible wavelength range.


Health-Related Quality of Life, Functioning, and Physical Symptoms of Adult Omani Colorectal Cancer Survivors.
Colorectal (CRC) survivors often experience physical and psychological symptoms affecting their health-related quality of life (HRQoL). This study aimed to identify factors impacting HRQoL-related functioning and physical symptoms among adult Omani CRC survivors. A cross-sectional study of 124 adult CRC survivors was conducted at the two main oncology referral hospitals in Oman. A validated Arabic version of the European Organization for Research and Treatment of Cancer-Quality of Life Questionnaire 30 was used to collect data. A total of 118 CRC survivors participated in the study (response rate: 95.2%). The mean age was 52 years and there were an equal number of male and female participants (n = 59 each; 50.0%). A total of 62 survivors (52.5%) had been diagnosed with CRC at stages III or IV. The overall score for global health was high (81.7). With regards to functioning, high mean scores were observed for domains of role (91.0) and social (90.7) functioning. In terms of symptoms, high mean scores were reported for constipation (25.4), insomnia (25.1), pain (20.1), and fatigue (18.9). Survivors under 60 years old (β=15.5, p=.004) and those with no comorbidities (β=16.0, p=.001) demonstrated better functional HRQoL. Being male was predictive of better functional HRQoL in the emotional (β=13.9, p<0.008), cognitive functioning (β=12.5, p=.013), role functioning (β=14.0, p=0.006) and physical functioning (β=17.8, p<0.001) domains. Healthcare professionals in Oman should implement measures to enhance the HRQoL of CRC survivors, particularly women and those with coexisting morbidities.