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Paralytic shellfish toxins (PST) in shellfish products have led to severe risks to human health. To monitor the risk, the Canadian Shellfish Sanitation Program has been collecting longitudinal PST measurements in blue mussel (Mytilus edulis) and soft-shell clam (Mya arenaria) samples in six coastal provinces of Canada. The spatial distributions of major temporal variation patterns were studied via Functional Principal Component Analysis. Seasonal increases in PST contamination were found to vary the most in terms of magnitude along the coastlines, which provides support for location-specific management of the time-sensitive PST contamination. In British Columbia, the first functional principal component (FPC1) indicated the variance among the magnitudes, while FPC2 indicated the seasonality of the PST levels. The temporal variations tended to be positively correlated with the abundance of dianoflagellates Alexandrium spp., and negatively with precipitation and inorganic nutrients. These findings indicate the underlying mechanism of PST variation in various geographical settings. In New Brunswick, Prince Edward, and Nova Scotia, the top FPCs indicated that the PST contamination differed mostly in the seasonal increase of the PST level during summer. • Multi-decade Paralytic Shellfish Toxins (PST) in Canada were analyzed. • Shellfish harvesting sites differed the most in seasonal elevation of PST levels. • The spring-to-autumn period of increased PST was found in BC, QC, NB, and NS. • Rainfall, Alexandrium spp., nitrate/nitrite, and phosphate correlated with PST. • Higher seasonal increases tended to occur at locations of strong water mixing. [ABSTRACT FROM AUTHOR]

In this experiment, three different producing methods, that is pre-fermentation, refrigeration (4 ℃, 24 h) and then re-fermentation (C1), refrigeration (4 ℃, 24 h) and then fermentation (C2), fermentation and then refrigeration (4 ℃, 24 h) (C3), were conducted to compare their effects on physicochemical and fermentation properties of refrigerated dough. The results showed that under refrigerated storage, the number of yeast cells decreased slightly, while yeast activity in C2 dough was higher than that in C1 and C3 dough. Dough refrigeration (4 ℃) led to a decrease in reducing sugar, ethanol, and glycerol content, an increase in the glutathione (GSH) content, and a decrease in amylase activity. In refrigerated dough, the α-helix became irregularly curled, the degree of water binding to other macromolecules weakened, and the proportion of free water increased. Compared to the control group, the content of total sulfhydryl group, disulfide bond, hydrogen bonds, and hydrophobic interactions increased in the refrigerated dough, the tensile force and displacement of C1 and C3 refrigerated dough were higher than C2 dough, G′ and G" in C1 dough were higher than that in C2 and C3 dough. Yeast with interrupted growth was more vulnerable to low-temperature stress, low temperature affected the quality of dough by affecting the activity of yeast, yeast metabolites, and the intermolecular force of dough. • Yeast with interrupted growth was more vulnerable to low temperature stress. • In refrigerated dough, binding degree of water with other macromolecules weakened. • Dough rheological property was affected by yeast activity and intermolecular force. [ABSTRACT FROM AUTHOR]

Background: Whole-genome gene expression analysis has been successfully utilized to diagnose, prognosticate, and identify potential therapeutic targets for high-risk cardiovascular diseases. However, the feasibility of this approach to identify outcome-related genes and dysregulated pathways following first-time myocardial infarction (AMI) remains unknown and may offer a novel strategy to detect affected expressome networks that predict long-term outcome. Methods and results: Whole-genome expression microarray on blood samples from normal cardiac function controls (n=21) and first-time AMI patients (n=31) within 48-hours post-MI revealed expected differential gene expression profiles enriched for inflammation and immune-response pathways. To determine molecular signatures at the time of AMI associated with long-term outcomes, transcriptional profiles from sub-groups of AMI patients with (n=5) or without (n=22) any recurrent events over an 18-month follow-up were compared. This analysis identified 559 differentially-expressed genes. Bioinformatic analysis of this differential gene-set for associated pathways revealed 1) increasing disease severity in AMI patients is associated with a decreased expression of genes involved in the developmental epithelial-to-mesenchymal transition pathway, and 2) modulation of cholesterol transport genes that include ABCA1, CETP, APOA1, and LDLR is associated with clinical outcome. Conclusion: Differentially regulated genes and modulated pathways were identified that were associated with recurrent cardiovascular outcomes in first-time AMI patients. This cell-based approach for risk stratification in AMI could represent a novel, non-invasive platform to anticipate modifiable pathways and therapeutic targets to optimize long-term outcome for AMI patients and warrants further study to determine the role of metabolic remodeling and regenerative processes required for optimal outcomes. [ABSTRACT FROM AUTHOR]

Abstract: The detailed molecular mechanisms underlying pathogenesis of various vascular diseases such as atherosclerosis are not fully understood in type-2 diabetes. The present study was designed to investigate whether insulin regulates KCa3.1 channels and participates in vasculopathy in type-2 diabetes. A rat model with experimental insulin-resistant type-2 diabetes was used for detecting pathological changes in the aorta wall, and cultured vascular smooth muscle cells (VSMCs) were employed to investigate the regulation of KCa3.1 channels by insulin and roles of KCa3.1 channels in cell migration and proliferation using molecular biology and electrophysiology. Early pathological changes were observed and expression of KCa3.1 channels increased in the aorta wall of the type 2 diabetic rats. KCa3.1 channel mRNA, protein levels and current density were greatly enhanced in cultured VSMCs treated with insulin, and the effects were countered in the cells treated with the ERK1/2 inhibitor PD98059, but not the p38-MAPK inhibitor SB203580. In addition, insulin stimulated cell migration and proliferation in cultured VSMCs, and the effects were fully reversed in the cells treated with the KCa3.1 blocker TRAM-34 or PD98059, but not SB203580. These results demonstrate the novel information that insulin increases expression of KCa3.1 channels by stimulating ERK1/2 phosphorylation thereby promoting migration and proliferation of VSMCs, which likely play at least a partial role in the development of vasculopathy in type-2 diabetes. [Copyright &y& Elsevier]

With the increasing concern over the potential pollution from farm wastes, there is a need for rapid and robust methods that can evaluate livestock manure nutrient content. Physicochemical models which related cattle manure nutrient content (ammonium nitrogen, AN; total potassium, TK; total nitrogen, TN; total phosphorus, TP) to its physicochemical properties (specific gravity, dry matter, electrical conductivity, pH) have been reported by previous researchers. This study reviewed previous physicochemical models and compiled the observed data drawn from a wide selection of sources to validate various physicochemical models. Several statistical parameters, including the coefficient of determination R 2, the modelling efficiency statistic, the mean squared error of prediction, the mean bias, and the linear bias, were calculated to evaluate model performance on the data sets. The results showed that the relationship (AN=0.136EC−0.523) developed by , provided satisfactory predictions for AN with R 2 =0.75. The equation (TP=0.01DM+0.057) developed by gave reasonable prediction for TP with R 2 =0.77. The corrected TK model (TK=0.184EC+0.042) greatly reduced systemic errors and hence improved TK prediction with R 2 =0.87. Compared with prediction models for other nutrient contents, TN physicochemical models provided far from satisfactory predictions but equations used by was the best (R 2 =0.62). [Copyright &y& Elsevier]

Prediction models for rapidly determining the nutrient content of pig manures will become important as environmental regulations increase the requirements for this information. Physicochemical models which related pig manure nutrient content (ammonium nitrogen, AN; total potassium, TK; total nitrogen, TN; total phosphorus, TP) to its physicochemical properties (specific gravity, dry matter, electrical conductivity, pH) have been reported by previous researchers. This study reviewed previous physicochemical models and compiled the observed data drawn from a wide selection of sources to validate various physicochemical models. Several statistical parameters, including the coefficient of determination (R 2), the modelling efficiency statistic, the mean squared error of prediction, the mean bias, and the linear bias, were calculated to evaluate model performance on the datasets. The results showed that the relationship developed by Scotford et al. [Scotford I M; Cumby T R; White R P; Carton O T; Lorenz F; Hatterman U, et al. (1998b). Estimation of the nutrient value of agricultural slurries by measurement of physical and chemical properties. Journal of Agricultural Engineering Research, 71(3), 291–305], can provide satisfactory predictions for AN with R 2 =0.89. The equations developed by Scotford et al. [Scotford I M; Cumby T R; White R P; Carton O T; Lorenz F; Hatterman U, et al. (1998b). Estimation of the nutrient value of agricultural slurries by measurement of physical and chemical properties. Journal of Agricultural Engineering Research, 71(3), 291–305] and Zhu et al. [Zhu J; Zhang Z; Ndegwa P M (2003). Using a soil hydrometer to measure the nitrogen and phosphorus contents in pig slurries. Biosystems Engineering, 85(1), 121–128], gave reasonable prediction for TK and TP with R 2 =0.88 and 0.78, respectively. Compared with prediction models for other nutrient contents, TN physicochemical models provided far from satisfactory predictions but equations used by Martinez-Suller et al. [Martinez-Suller L; Azzellino A; Provolo G (2008). Analysis of livestock slurries from farms across northern Italy: relationship between indicators and nutrient content. Biosystems Engineering, 99(4), 540–552], were the best (R 2 =0.73). These findings demonstrated that physicochemical model may be an appropriate tool to predict AN, TK, and TP content of pig manure, however, further work is needed to develop and validate TN physicochemical models. [Copyright &y& Elsevier]

With increasing concern over the potential pollution from farm wastes, there is a need for rapid and robust methods that can analyse animal manure. In order to evaluate rapid testing methods based on the relationship between layer manure composition (ammonium nitrogen, total potassium, total nitrogen, total phosphorus, iron, copper, zinc, magnesium and sodium) and physicochemical properties (specific gravity, electrical conductivity, pH), diverse layer manure samples (n =105) were used. Relationships were investigated using linear regression and artificial neural networks (ANNs). The performance of a neural network-based model was compared with a linear regression-based model using the same observed data. It was found that ANN model consistently gives better predictions. Based on the results of this study, ANNs appear to be a promising technique for predicting layer manure composition. [Copyright &y& Elsevier]

Abstract: Fabry disease, an X-linked systemic vasculopathy, is caused by a deficiency of α-galactosidase A resulting in globotriaosylceramide (Gb3) storage in cells. The pathogenic role of Gb3 in the disease is not known. Based on previous work, we tested the hypothesis that accumulation of Gb3 in the vascular endothelium of Fabry disease is associated with increased production of reactive oxygen species (ROS) and increased expression of cell adhesion molecules. Gb3-loading resulted in increased intracellular ROS production in cultured vascular endothelial cells in a dose-dependent manner. Increased Gb3 also induced expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin. Reduction of endogenous Gb3 by treatment of the cells with an inhibitor of glycosphingolipid synthase or α-galactosidase A led to decreased expression of adhesion molecules. Plasma from Fabry patients significantly increased ROS generation in endothelial cells when compared with plasma from non-Fabry controls. This effect was not influenced by reduction of intracellular Gb3. This study provided direct evidence that excess intracellular Gb3 induces oxidative stress and up-regulates the expression of cellular adhesion molecules in vascular endothelial cells. In addition, other factors in patient’s plasma may also contribute to oxidative stress in Fabry vascular endothelial cells. [Copyright &y& Elsevier]

Abstract: We report a facile solvothermal method for producing Co/C microspheres composed of nanosheet structures in a mixture of water and ethylene glycol. The morphology and cobalt loading of the Co/C microspheres can be controlled by adjusting the solvent composition. The Co/C microspheres obtained in the water–ethylene glycol mixture and calcined at 700 °C were very active for the hydrogenation of chloronitrobenzene (CNB), with 98% CNB conversion and 97% selectivity to o-chloroaniline at 2.0 MPa and 140 °C. In the hydrogenation of p-CNB and m-CNB, the catalyst showed a conversion of ⩾97% and a selectivity of 99% to the corresponding chloroanilines. [Copyright &y& Elsevier]

Abstract: Fabry disease is an inborn error of glycosphingolipid catabolism resulting from a deficiency of lysosomal enzyme α-galactosidase A. The major clinical manifestations of the disease, such as stroke, cardiac dysfunction, and renal impairment, are thought to be caused by vasculopathy due to progressive accumulation of globotriaosylceramide in vascular endothelial cells. The pathogenesis of the vasculopathy has not been elucidated. Since in vitro studies using primary endothelial cells are hampered by the limited lifespan of these cells, the availability of cultured endothelial cells with an extended lifespan is critical for the study of the vasculopathy of Fabry disease. We therefore generated an endothelial cell line from a Fabry hemizygote by introduction of human telomerase reverse transcriptase gene. The cell line has markedly extended lifespan compared to parental primary cells. The cells stably express many key markers of endothelial cells such as von Willebrand factor, CD31, CD34, and endothelial nitric oxide synthase (eNOS) and retain functional characteristics such as uptake of acetylated low-density lipoprotein, responsiveness to angiogenic growth factors, up-regulation of eNOS production upon extracellular stimuli, and formation of tube-like structures on Matrigel basement membrane matrix. The cells show significantly reduced activity of α-galactosidase A compared with primary endothelial cells from normal individuals and accumulate globotriaosylceramide in lysosomes. This cell line will provide a useful in vitro model of Fabry disease and will facilitate systematic studies to investigate pathogenic mechanisms and explore new therapeutic approaches for Fabry disease. [Copyright &y& Elsevier]

Abstract: Globoid cell leukodystrophy (GLD, Krabbe disease) is a severe demyelinating disease caused by a genetic defect of β-galactocerebrosidase (GALC). To date treatment to GLD is limited to hematopoietic stem cell transplantation. Experimental approaches by means of gene therapy in twitcher mouse, an authentic murine model of human GLD, showed significant but only marginal improvements of the disease. To clarify whether the introduction of GALC could provide beneficial effects on the oligodendrocytes in GLD, we transduced twitcher oligodendrocytes by stereotactically injecting recombinant retrovirus encoding GALC-myc-tag fusion gene into the forebrain subventricular zone of neonatal twitcher mouse. In vivo effects of exogenous GALC on twitcher oligodendrocytes were studied histologically by combined immunostaining for the myc-epitope and the oligodendroglial specific marker, π form of glutathione-S-transferase, at around 40 days of age. We show here that GALC transduction led to dramatic morphological improvement of the twitcher oligodendrocytes comparing with those in untreated twitcher controls. This study provided direct in vivo evidence that GALC transduction could prevent or correct aberrant morphology of oligodendrocytes in GLD which may be closely related to the dysfunction and/or degeneration of oligodendrocytes and the demyelination in this disease. [Copyright &y& Elsevier]

It has been demonstrated that cellular and viral RNAs were packaged in the virions of human cytomegalovirus (CMV) and herpes simplex virus 1 (HSV 1), members of the Herpesviridae family, both of which are enveloped double-stranded DNA viruses. Here, we provide evidence suggesting that RNAs are packaged in the virions of porcine adenovirus type 3 (PAdV-3), which is a member of the Adenoviridae family, a non-enveloped double-stranded DNA virus. The RNAs packaged in PAdV-3 virions were enriched in the size range of 300–1000 bases long. By reverse transcription (RT) of RNAs isolated from purified PAdV-3 virions, PCR amplification, and DNA sequence analysis of PCR products, we determined the identities of some viral RNAs contained in PAdV-3 virions. The results indicated that the RNAs representing transcripts from E1A, E1B, DNA binding protein (DBP), DNA polymerase (POL), E4 and some of the late genes including pIIIA, pIII, pV, Hexon, 33 K, and fiber were detected from purified PAdV-3 virions. In contrast, we could not detect the RNAs representing transcripts of precursor terminal protein (pTP), 52 kDa, pX, or 100-kDa protein genes in purified virions. Because the transcripts of pIX, IVa2, E3, protease, pVI, pVII, and pVIII overlap with those of other genes in PAdV-3, we could not definitely conclude that RNAs representing these transcripts were packaged in virions although the expected DNA fragments were produced by RT-PCR in the RNAs isolated from purified virions. [Copyright &y& Elsevier]

We identified a bifunctional regulatory element located between nt 374 and 431 upstream of TATA box of porcine adenovirus (PAV) 3 E1A promoter. Deletion of the element dramatically reduced the steady-state level of E1A mRNA, but increased that of E1B, which lies immediately downstream of E1A. The mutant virus displayed defective replication at early times of infection, but replicated nearly as efficiently as wild-type PAV-3 at late times of infection. This defect was complemented with coinfecting wild-type virus in a mixed infection. The results indicated that the upstream activation sequences (UAS) of E1A overlap the upstream repression sequences (URS) of E1B, although both transcription units are transcribed from different promoters. [Copyright &y& Elsevier]

Encapsidation of adenovirus DNA involves specific interactions between cis-acting genomic DNA sequences and trans-acting proteins. The cis-acting packaging domain located near the left inverted terminal repeat is composed of a series of redundant but not functionally equivalent motifs. Such motifs are made up of the consensus sequence 5′-TTTGN8CG-3′ and 5′-TTTG/A-3′ in human adenovirus 5 (HAV-5) and canine adenovirus-2 (CAV-2), respectively. To gain comparative insight into adenovirus encapsidation, we examined the packaging domain of porcine adenovirus-3 (PAV-3). Using deletion mutants, we localized the PAV-3 packaging domain to 319 bp (nt 212 to 531), which contains six cis-acting elements. However, this domain does not contain the consensus motifs identified in HAV-5. In addition, consensus motif found in CAV-2 is present only once in PAV-3. Instead, PAV-3 packaging domain appears to contain AT/GC-rich sequences. The packaging motifs of PAV-3, which are functionally redundant but not equivalent, are located at the left end of the genome. [Copyright &y& Elsevier]

[Display omitted] Chirality dynamic tuning plays fundamental roles in chemistry, material science and biological system. Herein, a pair of azobenzene-bridged bis-tryptophan enantiomers (Azo-di- d / l -Trp) were designed and synthesized via simple reactions. With the fuel of glucono-δ-lactone (GdL), releasing protons during its hydrolysis, the alkaline solution of Azo-di- d / l -Trp gradually self-assembled into contrast chiral helical structures and displayed magnitude and mirror image of circular dichroism (CD) signals. While the chiral helices converted to CD silent nanoparticles when the azobenzene moiety isomerized from trans - to cis -form under UV irradiation. More importantly, this chiroptical switch, displaying reversible interconversion between chiral amplification and silent, can be smartly controlled via photoirradiation at various wavelengths. [ABSTRACT FROM AUTHOR]

The Lovelock theory of gravity is a natural extension of the Einstein's theory of general relativity to higher dimensions, which presents the criteria of general covariance and is free of ghosts. In this work, we consider a model that static spherically-symmetric black hole is surrounded by the modified Chaplygin gas with equation of state p = A ρ − B / ρ α . We find that the energy density of the modified Chaplygin gas can be characterized by four parameters. The black hole thermodynamical quantities, consisting of gravitational mass, temperature, entropy and heat capacity, as functions of horizon radius, are calculated. Concerning the thermodynamical stability of the black holes, we consider the third-order Lovelock gravity as a specific case and show in figures the heat capacity versus the horizon radius, observing that there exist a phase of thermodynamically stable small black hole and a phase of thermodynamically unstable large black hole for D = 7 , while an additional intermediate local unstable phase as well as an intermediate local stable phase appear for D = 8 with configuring properly the model parameters. The effects of the modified Chaplygin gas and Lovelock gravity on thermodynamical stability of the black holes are discussed. • Lovelock black holes surrounded by modified Chaplygin gas are studied. • D = 7 3-order case, there are a phase of stable small and a phase of unstable large BH. • D = 8 3-order case, additional local unstable/stable phases could intermediately appear. [ABSTRACT FROM AUTHOR]

In order to fully understand the carbon emission from different fuels in rural villages of China, especially in the typical atmospheric pollution areas. The characteristics of carbonaceous aerosols and carbon dioxide (CO 2) with its stable carbon isotope (δ13C) were investigated in six households, which two households used coal, two households used wood as well as two households used biogas and liquefied petroleum gas (LPG), from two rural villages in Fenwei Plain from March to April 2021. It showed that the fine particulate matter (PM 2.5) emitted from biogas and LPG couldn't be as lower as expected in this area. However, the clean fuels could relatively reduce the emissions of organic carbon (OC) and element carbon (EC) in PM 2.5 compare to the solid fuels. The pyrolyzed carbon (OP) accounted more total carbon (TC) in coal than the other fuels use households, indicating that more water-soluble OC existed, and it still had the highest secondary organic carbon (SOC) than the other fuels. Meantime, the coal combustions in the two villages had the highest CO 2 concentration of 527.6 ppm and 1120.6 ppm, respectively, while the clean fuels could effectively reduce it. The average δ13C values (−26.9‰) was much lighter than almost all the outdoor monitoring and similar to the δ13C values for coal combustion and vehicle emission, showing that they might be the main contributors of the regional atmospheric aerosol in this area. During the sandstorm, the indoor PM 2.5 mass and CO 2 were increasing obviously. The indoor cancer risk of PAHs for adults and children were greater than 1 × 10−6, exert a potential carcinogenic risk to human of solid fuels combustion in rural northern China. It is important to continue concern the solid fuel combustion and its health impact in rural areas. [Display omitted] • Biogas and liquefied petroleum gas (LPG) emitted high PM 2.5 as unexpected. • Biogas and LPG emitted lower PM 2.5 bounded PAHs and CO 2. • The CO 2 produced by coal burning was greater than other fuels, with lighter δ13C. • Sandstorm could aggravate the accumulation of indoor atmospheric CO 2. • Solid fuel users might face more serious cancer risks than biogas and LPG. [ABSTRACT FROM AUTHOR]

Abstract: Our study is the first to demonstrate the ability to generate iPS cells from a mouse model of Pompe disease. Initially, mouse tail tip fibroblasts were harvested from male, 8-week-old (GAA) knockout mice, and three reprogramming factors (Oct3/4, Sox2 and Klf4) were transfected into the isolated donor cells using a retroviral vector. These iPS cells also showed decreased levels of GAA enzymatic activity and strong positive staining with periodic acid-Schiff (indicating the accumulation of glycogen) and acid phosphatase (lysosomal activation marker). Pompe-iPS cells were differentiated into skeletal muscle cells in Matrigel®-coated plates. Spindle-shaped skeletal muscle cells were successfully generated from Pompe-iPS cells and showed spontaneous contraction and positive staining with the myosin heavy chain antibody. Electron microscopic analysis of the skeletal muscle cells showed typical morphological features, including Z-bands, I-bands, A-bands and H-bands, which were visible in wild-type and Pompe cells. Furthermore, Pompe skeletal muscle cells accumulated massive glycogen in lysosomes. This study indicates that the iPS and skeletal muscle cells generated in this study could also be a useful disease model for studies investigating the pathogenesis and treatment of skeletal muscle in Pompe disease. [Copyright &y& Elsevier]

Emerging evidence has linked autophagy to skin wound healing; however, the underlying cellular and molecular mechanisms remain poorly understood. The present study was designed to determine the role of autophagy in endothelial cell (EC)-mediated skin wound healing in mice. Autophagy-related gene (Atg7) in mouse ECs was inactivated by the Cre-loxP system under the control of an EC-specific VE-Cadherin (Cdh5) promoter (Atg7EC−/− mice). Full-thickness skin wounds were created on the dorsum of wild-type (WT), Cdh5-Cre+, floxed Atg7 (Atg7F/F), and Atg7EC−/− mice. Autophagic activity was determined by autophagic flux assay in the primary culture of ECs isolated from these mice. The wound re-epithelialization and angiogenesis was examined by histological analyses. The angiogenic activity of ECs was evaluated by tube formation assay in vitro. EC proliferation was examined by a cell count CCK-8 kit. EC-originated intercellular communication with dermal fibroblasts and keratinocytes was assessed by measuring the effect of EC conditional medium on the growth of keratinocytes and fibroblasts. The levels of VEGF, EGF, bFGF in EC conditional medium were measured by ELISA. Autophagy deficiency in ECs markedly enhanced the re-epithelialization and the wound closure during skin wound healing. However, it has minimal impact on angiogenesis in the wounded skin. Notably, autophagy deficiency in ECs did not affect their proliferation and migration or angiogenic activity per se but enhanced the EC conditional medium-induced proliferation and migration of keratinocytes and fibroblasts. These results demonstrate for the first time an inhibitory role of autophagy in the EC-originated paracrine regulation of skin wound healing. [ABSTRACT FROM AUTHOR]

Studies have proved the role of GAS5 in the development of different cancers. This study was undertaken to investigate the role and explore therapeutic implications of GAS5 in human cervical cancer. The results showed that GAS5 was significantly (p < 0.05) downregulated in human cervical cancer tissues. The results also showed that cervical cancer progresses with the suppression of GAS5 expression levels. Additionally, the expression of GAS5 was also significantly (p < 0.05) downregulated in human cervical cancer cell lines. Nonetheless, overexpression of GAS5 caused a remarkable decrease in the proliferation of C33A and HeLa cervical cancer cells. The decrease in the proliferation rate was attributed to the induction of apoptosis of C33A and HeLa cells which was accompanied with upregulation of Bax and suppression of Bcl-2. Additionally, GAS5 overexpression also promoted the arrest of C33A and HeLa cells at the G2/M check point of cell cycle via suppression of cyclin B1 and CDK1 expression. The transwell assays showed that GAS5 overexpression significantly (p < 0.05) inhibited the migration and invasion of the C33A and HeLa cervical cancer cells. The bioinformatics analysis as well as the dual luciferase assay showed GAS5 acts as a target of miR-135a. Interestingly, the expression of miR-135a was upregulated in the human cervical cancer cells and its suppression exerted growth inhibitory effects on the C33A and HeLa cells. However, silencing of GAS5 could nullify the effects of miR-135a suppression on the proliferation of C33A and HeLa cells. Taken together, the results of this study point towards the therapeutic implications of GAS5 in the treatment of cervical cancer. [ABSTRACT FROM AUTHOR]

• C. perfringens triggers classical phagocyte ETs formation. • Bacteria induce ETs release via ERK1/2-, P38 MAPK- and SOCE-dependent mechanisms. • A beneficial role of phagocyte ETs on pathogen control in vitro and in vivo. Clostridium perfringens (C. perfringens) is an important Gram-positive anaerobic spore-forming pathogen that provokes life-threatening gas gangrene and acute enterotoxaemia, although it colonizes as a component of the symbiotic bacteria in humans and animals. However, the mechanisms by which C. perfringens is cleared from the host remains poorly understood, thereby impeding the development of novel strategies for control this infection. Here, we uncover a beneficial effect of extracellular traps (ETs) formation on bacterial killing and clearance by phagocytes. C. perfringens strain ATCC13124, and wild-type isolates CP1 and CP3 markedly trigger ETs formation in macrophages and neutrophils. As expected, visualization of DNA decorated with histone, myeloperoxidase (MPO) and neutrophils elastase (NE) in C. perfringens -triggered classical ETs structures. Notably, the bacteria-induced ETs formation is an ERK1/2-, P38 MAPK-, store-operated calcium entry (SOCE)-, NADPH oxidase-, histone-, NE-, and MPO-dependent process, and is independent of LDH activity. Meanwhile, the defect of bactericidal activity is mediated by impairing ETs formation in phagocytes. Moreover, In vivo studies indicated that degradation of ETs by DNase I administration leads to a defect in the protection against experimental gas gangrene, with higher mortality rates, exacerbated tissue damage, and more bacterial colonization. Together, these results suggest that phagocyte ETs formation is essential for the host defense against C. perfringens infection. [ABSTRACT FROM AUTHOR]

Although exposure to ambient air pollution has been associated with mental disorder, little is known about its potential effects on children and adolescents, especially in Chinese population. We aimed to reveal the relationship of air pollutants with hospital outpatient visits for child and adolescence psychiatry (HOVCAP) in Shenzhen. A case-crossover study based on time-series data was applied, and a distributed lag non-linear model (DLNM) was used to evaluate the non-linear and delayed effects of 4 major air pollutants (NO 2 , PM 2.5 , SO 2 and O 3) on HOVCAP. Least absolute shrinkage and selection operator (LASSO) regression was used to control the multicollinearity between covariates and to filter variables. A total of 94,660 cases aged 3–18 were collected from 2014 to 2019 in the Mental Health Center of Shenzhen. Results of pollutants at mode value (M 0) showed that in the single lag effect result, when the average daily concentration of NO 2 at 24 μg/m3, there was a significant effect on HOVCAP over lag 1, lag 4 and lag 5, respectively. The cumulative RR of NO 2 M 0 value to the outpatient visits were 1.438 (1.137–1.818) over lag 0–2, 1.454 (1.120–1.887) over lag 0–3, 1.466 (1.084–1.982) over lag 0–4, 1.680 (1.199–2.354) over lag 0–5, 1.993 (1.369–2.903) over lag 0–6, and 2.069 (1.372–3.119) over lag 0–7. However, PM 2.5 , SO 2 , O 3 were not associated with HOVCAP over neither single lag effects nor cumulative effects. The RR values both shown an increase either when NO 2 increases by 10 units or when the maximum concentration of NO 2 is reached. Our study suggests that exposure to the normal air quality of NO2 in Shenzhen may associated with the risk of HOVCAP. However, PM2.5, SO2, O3 were not associated with HOVCAP. • A distributed lag non-linear model (DLNM) was used to evaluate the non-linear and delayed effects of pollutants on HOVCAP (hospital outpatient visits for child and adolescence psychiatry). • Exposure to the normal air quality of NO2 in Shenzhen may associated with the risk of HOVCAP. • The overall effect curve of NO2 on HOVCAP demonstrated an inverted U shape. [ABSTRACT FROM AUTHOR]

Chronic activation of the myocardial renin angiotensin system (RAS) elevates the local level of angiotensin II (Ang II) thereby inducing pathological cardiac hypertrophy, which contributes to heart failure. However, the precise underlying mechanisms have not been fully delineated. Herein we report a novel paracrine mechanism between cardiac fibroblasts (CF)s and cardiomyocytes whereby Ang II induces pathological cardiac hypertrophy. In cultured CFs, Ang II treatment enhanced exosome release via the activation of Ang II receptor types 1 (AT 1 R) and 2 (AT 2 R), whereas lipopolysaccharide, insulin, endothelin (ET)-1, transforming growth factor beta (TGFβ)1 or hydrogen peroxide did not. The CF-derived exosomes upregulated the expression of renin, angiotensinogen, AT 1 R, and AT 2 R, downregulated angiotensin-converting enzyme 2, and enhanced Ang II production in cultured cardiomyocytes. In addition, the CF exosome-induced cardiomyocyte hypertrophy was blocked by both AT 1 R and AT 2 R antagonists. Exosome inhibitors, GW4869 and dimethyl amiloride (DMA), inhibited CF-induced cardiomyocyte hypertrophy with little effect on Ang II-induced cardiomyocyte hypertrophy. Mechanistically, CF exosomes upregulated RAS in cardiomyocytes via the activation of mitogen-activated protein kinases (MAPKs) and Akt. Finally, Ang II-induced exosome release from cardiac fibroblasts and pathological cardiac hypertrophy were dramatically inhibited by GW4869 and DMA in mice. These findings demonstrate that Ang II stimulates CFs to release exosomes, which in turn increase Ang II production and its receptor expression in cardiomyocytes, thereby intensifying Ang II-induced pathological cardiac hypertrophy. Accordingly, specific targeting of Ang II-induced exosome release from CFs may serve as a novel therapeutic approach to treat cardiac pathological hypertrophy and heart failure. [ABSTRACT FROM AUTHOR]

Interendothelial junctions play an important role in the maintenance of endothelial integrity and the regulation of vascular functions. We report here that cationic amino acid transporter-1 (CAT-1) is a novel interendothelial cell adhesion molecule (CAM). We identified that CAT-1 protein localized at cell–cell adhesive junctions, similar to the classic CAM of VE-cadherin, and knockdown of CAT-1 with siRNA led to an increase in endothelial permeability. In addition, CAT-1 formed a cis -homo-dimer and showed Ca 2 + -dependent trans -homo-interaction to cause homophilic cell–cell adhesion. Co-immunoprecipitation assays showed that CAT-1 can associate with β-catenin. Furthermore, we found that the sub-cellular localization and function of CAT-1 are associated with cell confluency, in sub-confluent ECs CAT-1 proteins distribute on the entire surface and function as l -Arg transporters, but most of the CAT-1 in the confluent ECs are localized at interendothelial junctions and serve as CAMs. Further functional characterization has disclosed that extracellular l- Arg exposure stabilizes endothelial integrity via abating the cell junction disassembly of CAT-1 and blocking the cellular membrane CAT-1 internalization, which provides the new mechanisms for l- Arg paradox and trans -stimulation of cationic amino acid transport system (CAAT). These results suggest that CAT-1 is a novel CAM that directly regulates endothelial integrity and mediates the protective actions of l- Arg to endothelium via a NO-independent mechanism. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRs) dysregulation is a general feature of colorectal cancer (CRC) and other solid tumors, and is associated cancer progression. In the current study, we demonstrate that microRNA-101 (miR-101) inhibits CRC cells probably through down-regulating sphingosine kinase 1 (SphK1). Our results showed that exogenously expressing miR-101 inhibited CRC cell (HT-29 and HCT-116 lines) growth in vitro . At the molecular level, miR-101 dramatically down-regulated SphK1 mRNA and protein expression, causing pro-apoptotic ceramide production in above CRC cells. On the other hand, inhibition of miR-101 through expressing antagomiR-101 increased SphK1 expression to down-regulate ceramide level in HT-29 cells. miR-101 expression increased the in vitro anti-CRC activity of conventional chemo-agents: paclitaxel and doxorubicin. CRC cells with SphK1-shRNA knockdown showed similar phenotypes as the miR-101-expressed CRC cells, presenting with elevated level of ceramide and high sensitivity to paclitaxel or doxorubicin. In vivo, HCT-116 xenograft growth in severe combined immuno-deficient (SCID) mice was dramatically inhibited by over-expressing miR-101. Further, miR-101 enhanced paclitaxel-induced anti-HCT-116 activity in vivo . Together, these results indicate that miR-101 exerts its anti-CRC activities probably through down-regulating SphK1. [ABSTRACT FROM AUTHOR]

Ubiquitin proteasome system (UPS) consists of ubiquitin, ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), ubiquitin ligases (E3s), proteasomes, and deubiquitinating enzymes (DUBs). Ubiquitin, E1s, several E2s, E3s, and proteasomes play an important role in the regulation of cardiac homeostasis and dysfunction; however, less is known about the role of DUBs in the heart. Here, we uncovered a crucial role of cyclindromatosis (CYLD), a DUB, in mediating cardiac maladaptive remodeling and dysfunction. CYLD expression was dramatically upregulated in the cardiomyocytes of hypertrophic and failing human and murine hearts. Knockout of CYLD improved survival rate and alleviated cardiac hypertrophy, fibrosis, apoptosis, oxidative stress, and dysfunction in mice that were subjected to sustained pressure overload induced by transverse aortic constriction. Deep sequencing and gene array analyses revealed that the most dramatically changed genes are those involving in the free radical scavenging pathway and cardiovascular disease, including fos, jun, myc, and nuclear factor erythroid-2 related factor 2 (Nrf2) in the heart. Moreover, knockdown of CYLD enhanced mitogen-activated protein kinase (MAPK) ERK- and p38-mediated expression of c-jun, c-fos, and c-myc, which govern Nrf2 expression in cardiomyocytes. The CYLD deficiency-induced suppression of reactive oxygen species (ROS) formation, death and hypertrophy in cardiomyocytes was blocked by additional knockdown of Nrf2. Taken together, our findings demonstrate for the first time that CYLD mediates cardiac maladaptive remodeling and dysfunction, most likely via enhancing myocardial oxidative stress in response to pressure overload. At the molecular level, CYLD interrupts the ERK- and p38-/AP-1 and c-Myc pathways to suppress Nrf2-operated antioxidative capacity, thereby enhancing oxidative stress in the heart. [ABSTRACT FROM AUTHOR]

Recently GLP-1 was found to have cardioprotective effects independent of those attributable to tight glycemic control. Methods and results We employed ultrasound targeted microbubble destruction (UTMD) to deliver piggybac transposon plasmids encoding the GLP-1 gene with a nuclear localizing signal to rat hearts with adriamycin cardiomyopathy. After a single UTMD treatment, overexpression of transgenic GLP-1 was found in nuclei of rat heart cells with evidence that transfected cardiac cells had undergone proliferation. UTMD-GLP-1 gene therapy restored LV mass, fractional shortening index, and LV posterior wall diameter to nearly normal. Nuclear overexpression of GLP-1 by inducing phosphorylation of FoxO1-S256 and translocation of FoxO1 from the nucleus to the cytoplasm significantly inactivated FoxO1 and activated the expression of cyclin D1 in nuclei of cardiac muscle cells. Reversal of adriamycin cardiomyopathy appeared to be mediated by dedifferentiation and proliferation of nuclear FoxO1-positive cardiac muscle cells with evidence of embryonic stem cell markers (OCT4, Nanog, SOX2 and c-kit), cardiac early differentiation markers (NKX2.5 and ISL-1) and cellular proliferation markers (BrdU and PHH3) after UTMD with GLP-1 gene therapy. Conclusions Intranuclear myocardial delivery of the GLP-1gene can reverse established adriamycin cardiomyopathy by stimulating myocardial regeneration. [ABSTRACT FROM AUTHOR]

Abstract: Nuclear factor erythroid-2 related factor 2 (Nrf2) is a master transcription factor that controls the basal and inducible expression of a battery of antioxidant genes and other cytoprotective phase II detoxifying enzymes. While knockout of Nrf2 exaggerates cardiac pathological remodeling and dysfunction in diverse pathological settings, pharmacological activation of Nrf2 protects against cardiomyocyte injury and cardiac dysfunction. In contrast, there is also a concern that the chronic activation of Nrf2 secondary to oxidative stress is a contributing mechanism for the reductive stress-mediated heart failure. However, a direct link between cardiac specific activation of Nrf2 and cardiac protection or dysfunction in vivo remains to be established. Therefore, we investigated the effect of cardiomyocyte-specific transgenic activation of Nrf2 (Nrf2ctg) on cardiac pathological remodeling and dysfunction. We found that the cardiomyocyte-specific activation of Nrf2 suppressed myocardial oxidative stress as well as cardiac apoptosis, fibrosis, hypertrophy, and dysfunction in a setting of sustained pressure overload induced by transverse aortic arch constriction (TAC) in mice. Notably, the constitutive activation of Nrf2 increased the steady level of autophagosomes while decreasing the ubiquitinated protein aggregates in the heart after TAC. Nrf2 gene gain- and loss-of-function approaches revealed that Nrf2 enhances autophagosome formation and autophagic flux in cardiomyocytes. Unexpectedly, while Nrf2 minimally regulated apoptosis, it suppressed significantly the proteotoxic necrosis in cardiomyocytes. In addition, Nrf2 attenuated the proteocytotoxicity presumably via enhancing autophagy-mediated clearance of ubiquitinated protein aggregates in cardiomyocytes. Taken together, we demonstrated for the first time that cardiac specific activation of Nrf2 suppresses cardiac maladaptive remodeling and dysfunction most likely by enhancing autophagic clearance of toxic protein aggregates in the heart. [Copyright &y& Elsevier]

Highlights: [•] Dh404 suppresses the expression of a selected set of pro-inflammatory cytokines in inflamed macrophages via activating Nrf2. [•] Dh404 activates Nrf2 while keeping Keap1 function intact in macrophages. [•] Dh404 minimally regulates NF-κB pathway in macrophages. [ABSTRACT FROM AUTHOR]

Abstract: Interleukin 6 (IL-6) is a pivotal cytokine that regulates extracellular matrix metabolism by ameliorating the modification of collagen content, important in fibrous caps of atherosclerotic plaque. Prolyl-4-hydroxylase α1 (P4Hα1) is a key intracellular enzyme required for synthesis of collagen in animals. We investigated the relationship of IL-6 and P4Hα1 in atherosclerosis-prone mice and human aortic smooth muscle cells (HASMCs). Apolipoprotein E (ApoE)−/− mice were fed a high-fat diet and a perivascular constrictive silica collar was placed on the right common carotid artery to induce atherosclerotic lesions, then mice were divided into two groups for transfection with empty lentivirus or IL-6 lentivirus. HASMCs were transfected with small interfering RNA or treated with recombinant human IL-6. IL-6 significantly downregulated collagen, P4Hα1 and smooth muscle cell contents in atherosclerotic mouse arteries. Macrophage and lipid contents in the atherosclerotic area were significantly increased with IL-6 treatment. IL-6 significantly downregulated P4Hα1 expression in HASMCs through an RAF-MEK1/2-ERK1/2 mitogen-activated protein kinase (MAPK) pathway, and c-Jun was involved in the process. Our findings highlight IL-6 destabilize atherosclerotic plaques in mice by downregulating P4Hα1 via an RAF-MEK1/2-ERK1/2 MAPK and c-Jun pathway. [Copyright &y& Elsevier]

Abstract: CYLD, a deubiquitinating enzyme (DUB), is a critical regulator of diverse cellular processes, ranging from proliferation and differentiation to inflammatory responses, via regulating multiple key signaling cascades such as nuclear factor kappa B (NF-κB) pathway. CYLD has been shown to inhibit vascular lesion formation presumably through suppressing NF-κB activity in vascular cells. However, herein we report a novel role of CYLD in mediating pro-inflammatory responses in vascular smooth muscle cells (VSMCs) via a mechanism independent of NF-κB activity. Adenoviral knockdown of Cyld inhibited basal and the tumor necrosis factor alpha (TNFα)-induced mRNA expression of pro-inflammatory cytokines including monocyte chemotactic protein-1 (Mcp-1), intercellular adhesion molecule (Icam-1) and interleukin-6 (Il-6) in rat adult aortic SMCs (RASMCs). The CYLD deficiency led to increases in the basal NF-κB transcriptional activity in RASMCs; however, did not affect the TNFα-induced NF-κB activity. Intriguingly, the TNFα-induced IκB phosphorylation was enhanced in the CYLD deficient RASMCs. While knocking down of Cyld decreased slightly the basal expression levels of IκBα and IκBβ proteins, it did not alter the kinetics of TNFα-induced IκB protein degradation in RASMCs. These results indicate that CYLD suppresses the basal NF-κB activity and TNFα-induced IκB kinase activation without affecting TNFα-induced NF-κB activity in VSMCs. In addition, knocking down of Cyld suppressed TNFα-induced activation of mitogen activated protein kinases (MAPKs) including extracellular signal-activated kinases (ERK), c-Jun N-terminal kinase (JNK), and p38 in RASMCs. TNFα-induced RASMC migration and monocyte adhesion to RASMCs were inhibited by the Cyld knockdown. Finally, immunochemical staining revealed a dramatic augment of CYLD expression in the injured coronary artery with neointimal hyperplasia. Taken together, our results uncover an unexpected role of CYLD in promoting inflammatory responses in VSMCs via a mechanism involving MAPK activation but independent of NF-κB activity, contributing to the pathogenesis of vascular disease. [Copyright &y& Elsevier]

Abstract: In this study, we evaluated four methods to separate and purify Toxoplasma gondii tachyzoites from in vivo and in vitro culture systems, including trypsin digestion, purification with a 3-μm filter, CF-11 cellulose purification, and Percoll purification. Our results indicate that both purification with a 3-μm filter and CF11 cellulose purification methods remove leukocytes or HeLa cells, and can therefore be used as candidate methods for the purification of in vivo and in vitro culture products. Trypsin digestion had a high tachyzoite recovery rate, but 22.35% of leukocytes and 69.64% of HeLa cells remained in the purified products. Percoll solution [30% (v/v)] also had a high tachyzoite recovery rate, but 3.44% of leukocytes and 61.61% of HeLa cells remained in the purified products. The 40% Percoll solution was also a candidate method for purifying tachyzoites from in vivo culture products, with a 65.45% tachyzoite recovery rate and without leukocytes. [Copyright &y& Elsevier]

Abstract: Avian polyomavirus (APV) causes a fatal, multi-organ disease among several bird species. Using cryogenic electron microscopy and other biochemical techniques, we investigated the structure of APV and compared it to that of mammalian polyomaviruses, particularly JC polyomavirus and simian virus 40. The structure of the pentameric major capsid protein (VP1) is mostly conserved; however, APV VP1 has a unique, truncated C-terminus that eliminates an intercapsomere-connecting β-hairpin observed in other polyomaviruses. We postulate that the terminal β-hairpin locks other polyomavirus capsids in a stable conformation and that absence of the hairpin leads to the observed capsid size variation in APV. Plug-like density features were observed at the base of the VP1 pentamers, consistent with the known location of minor capsid proteins VP2 and VP3. However, the plug density is more prominent in APV and may include VP4, a minor capsid protein unique to bird polyomaviruses. [Copyright &y& Elsevier]

Abstract: The continued spread of a highly pathogenic avian influenza (HPAI) H5N1 virus among poultry and wild birds has posed a potential threat to human public health. An influenza pandemic happens, when a new subtype that has not previously circulated in humans emerges. Almost all of the influenza pandemics in history have originated from avian influenza viruses (AIV). Birds are significant reservoirs of influenza viruses. In the present study, we performed a survey of avian influenza virus in ostriches and H5N1 virus (A/Ostrich/SuZhou/097/03, China097) was isolated. This H5N1 virus is highly pathogenic to both chickens and mice. It is also able to replicate in the lungs of, and to cause death in, BALB/c mice following intranasal administration. It forms plaques in chicken embryo fibroblast (CEF) cells in the absence of trypsin. The hemagglutinin (HA) gene of the virus is genetically similar to A/Goose/Guangdong/1/96(H5N1) and belongs to clade 0. The HA sequence contains multiple basic amino acids adjacent to the cleavage site, a motif associated with HPAI viruses. More importantly, the existence of H5N1 isolates in ostriches highlights the potential threat of wild bird infections to veterinary and public health. [Copyright &y& Elsevier]

Abstract: Objective: The accumulation of lipids in macrophages contributes to the development of atherosclerosis. Strategies to reduce lipid accumulation in macrophages may have therapeutic potential for preventing and treating atherosclerosis and cardiovascular complications. The antidiabetic drug metformin has been reported to reduce lipid accumulation in adipocytes. In this study, we examined the effects of metformin on lipid accumulation in macrophages and investigated the mechanisms involved. Methods and results: We observed that metformin significantly reduced palmitic acid (PA)-induced intracellular lipid accumulation in macrophages. Metformin promoted the expression of carnitine palmitoyltransferase I (CPT-1), while reduced the expression of fatty acid-binding protein 4 (FABP4) which was involved in PA-induced lipid accumulation. Quantitative real-time PCR showed that metformin regulates FABP4 expression at the transcriptional level. We identified forkhead transcription factor FOXO1 as a positive regulator of FABP4 expression. Inhibiting FOXO1 expression with FOXO1 siRNA significantly reduced basal and PA-induced FABP4 expression. Overexpression of wild-type FOXO1 and constitutively active FOXO1 significantly increased FABP4 expression, whereas dominant negative FOXO1 dramatically decreased FABP4 expression. Metformin reduced FABP4 expression by promoting FOXO1 nuclear exclusion and subsequently inhibiting its activity. Conclusions: Taken together, these results suggest that metformin reduces lipid accumulation in macrophages by repressing FOXO1-mediated FABP4 transcription. Thus, metformin may have a protective effect against lipid accumulation in macrophages and may serve as a therapeutic agent for preventing and treating atherosclerosis in metabolic syndrome. [Copyright &y& Elsevier]

Abstract: Objective: Atherosclerosis is a chronic inflammatory disease regulated by T lymphocyte subsets. Recently, CD4+ CD25+ Foxp3+ regulatory T (Treg) cells and Th17 cells have been described as two distinct subsets and have the opposite effects on autoimmunity. Clinical observation has revealed that the Th17/Treg imbalance exists in patients with acute coronary syndrome. We investigated whether the Th17/Treg functional imbalance existed during atherogenesis in ApoE−/− mice. Methods and results: Th17/Treg functions at different levels including cell frequencies, related cytokine secretion and key transcription factors were investigated comparatively between ApoE−/− mice and their age-matched C57BL/6J mice. The results demonstrated that ApoE−/− mice revealed significantly increased secretion of Th17 related cytokines (IL-17 and IL-6) and expression of transcription factor (RORγt) levels and obviously decreased number in Treg cells, secretion of Treg related cytokines (TGF-β1) and expression of transcription factor (Foxp3) levels as compared with age-matched C57BL/6J mice. Th17 related mediators reached their maximum expression values at the early stage (8–16weeks of age) in ApoE−/− mice, and then followed by continuous depression of their expression levels. Meanwhile, the expression of Treg related mediators was much lower in ApoE−/− mice than in their age-matched wild-type littermates. Conclusions: Th17/Treg functional imbalance exists during atherogenesis in ApoE−/− mice, suggesting a potential role of Th17/Treg imbalance in the formation and progression of atherosclerosis. [Copyright &y& Elsevier]

Background: The Accreditation Council of Graduate Medical Education mandated an 80-h/wk resident physician work-hour restriction on July 1, 2003. The objective of this study was to evaluate the impact of the resident work-hour restriction on outcomes of cardiac operations. Materials and Method: We reviewed records of 1562 patients who underwent cardiac operations at our institution between 1997 and 2007, and we compared outcomes of operations performed before July 1, 2003 (pre-reform, n =777) and those performed after July 1, 2003 (post-reform, n =785). Multivariate analysis with logistic regression was used to test for the independent effects of the resident work-hour reform by controlling for patient-specific confounding factors. Results: Post-reform patients had a significantly lower 30-d mortality rate (1.8% versus 3.9%; P =0.01) and a slightly lower 6-month mortality rate (4.5% versus 6.3%; P =0.12) than pre-reform patients. Multivariate analysis revealed that the post-reform patients had significantly lower adjusted 30-d mortality (odds ratio, 0.37; 95% CI, 0.18–0.75; P =0.006) and 6-mo mortality (odds ratio, 0.56; 95% CI, 0.34–0.91; P =0.02) than the pre-reform patients. Conclusions: Cardiac operations performed after the resident work-hour restriction went into effect were associated with significantly lower adjusted 30-d and 6-mo mortality rates than were operations performed before the work-hour restriction became effective. [Copyright &y& Elsevier]

Abstract: Oxidative stress induced by hyperglycemia is a key factor in the development of cardiovascular diseases in diabetes. Thioredoxin (Trx) system, a major thiol antioxidant system, regulates the reduction of intracellular reactive oxygen species (ROS). In this study, we demonstrated that high glucose significantly increased intracellular ROS levels in human aortic endothelial cells (HAECs). Additionally, high glucose reduced the antioxidant activity of thioredoxin. To investigate the mechanisms involved, we found that glucose enhanced the expression of thioredoxin interacting protein (Txnip), a Trx inhibitory protein, through p38 mitogen-activated protein kinase (MAPK). We also showed that glucose regulated Txnip at transcription level and p38 MAPK and forkhead box O1 transcriptional factor (FOXO1) were involved in the process. Taken together, upregulation of Txnip and subsequent impairment of thioredoxin antioxidative system through p38 MAPK and FOXO1 may represent a novel mechanism for glucose-induced increase in intracellular ROS. [Copyright &y& Elsevier]

Abstract: HMG-CoA reductase inhibitors and histone deacetylases (HDACs) inhibitors have been shown to induce apoptosis in a variety of cells, which could potentially be used as an anticancer therapy in addition to the designated applications. In the present study, we explored the possible synergistic pro-apoptotic effects and the underlying mechanisms when the two classes of inhibitors were combined. Exposure of HeLa cells to the combined treatment of mevastatin (an inhibitor of HMG-CoA reductase) and trichostatin A (TSA) (an inhibitor of HDACs) synergistically induced apoptosis. Mevastatin treatment transcriptionally and translationally up-regulated RhoA expression in the cells by negative feedback mechanism. While TSA enhanced mevastatin-induced RhoA up-regulation, more importantly, it also accelerated mevastatin-mediated depletion of membrane-bound (geranylgeranylated) RhoA. Moreover, TSA treatment down-regulated protein geranylgeranyl transferase-I (GGTase-I) β subunit expression, which is one of the key enzymes for protein geranylgeranylation. Taken together, TSA down-regulated GGTase-I β expression, hence enhanced the statin-induced depletion of geranylgeranylated RhoA, which could be an important mechanism for the synergistic induction of the apoptosis. [Copyright &y& Elsevier]

Abstract: Asymmetric dimethylarginine (ADMA), an inhibitor of nitric oxide synthase (NOS), has been identified as a new and emerging contributor to, or marker for, cardiovascular risk. The ADMA-mediated regulation of nitric oxide (NO) production is determined by the quantitative bioavailability of intracellular and extracellular ADMA. Dimethylarginine dimethylaminohydrolase (DDAH), which is ubiquitously expressed in tissues, especially liver and kidney, converts the majority of the ADMA to citrulline. In this review, we discuss a new regulatory mechanism for the metabolism of ADMA in which l-arginine acts as a competitive inhibitor of DDAH activity. This novel regulatory pathway is consistent with ADMA contributing to cardiovascular risk when levels are increased but not when levels are within the normal range. The pathway then has a physiological role in the regulation of NO production by preventing overproduction of NO. The regulatory role of l-arginine on ADMA may explain the unexpected outcomes in some l-arginine supplementation studies. This paper also reviews associations between the metabolism of ADMA and insulin resistance, smoking and homocysteine which are all associated with an increased risk of vascular disease. [Copyright &y& Elsevier]

Abstract: Lumazine synthases have been observed in the form of pentamers, dimers of pentamers, icosahedral capsids consisting of 60 subunits and larger capsids with unknown molecular structure. Here we describe the analysis of the assembly of native and mutant forms of lumazine synthases from Bacillus subtilis and Aquifex aeolicus at various pH values and in the presence of different buffers using small angle X-ray scattering and electron microscopy. Both wild-type lumazine synthases are able to form capsids with a diameter of roughly 160 Å and larger capsids with diameters of around 300 Å. The relative abundance of smaller and larger capsids is strongly dependent on buffer and pH. Both forms can co-exist and are in some cases accompanied by other incomplete or deformed capsids. Several mutants of the B. subtilis lumazine synthase, in which residues in or close to the active site were replaced, as well as an insertion mutant of A. aeolicus lumazine synthase form partially or exclusively larger capsids with a diameter of about 300 Å. The mutations also reduce or inhibit enzymatic activity, suggesting that the catalytic function of the enzyme is tightly correlated with its quaternary structure. The data show that multiple assembly forms are a general feature of lumazine synthases. [Copyright &y& Elsevier]

Abstract: Inhibition of histone deacetylases by trichostatin A (TSA) has pleiotropic effects on gene expression. We demonstrated that at low dose (0.1μg) TSA increased the eNOS mRNA levels, which was followed by a time- and dose-dependent down-regulation. Cycloheximide, a protein synthesis inhibitor, completely abolished TSA-induced decrease in eNOS expression, indicating that new protein synthesis is required for the inhibiting effect. Mevastatin—an inhibitor HMG-CoA reductase and geranylgeranylation reaction dose-dependently antagonized TSA-induced reduction. This mevastatin-mediated antagonism was completely abolished by geranylgeranylpyrophosphate, suggesting that geranylgeranyl modification is needed to activate the eNOS mRNA destabilizing factor—a mechanism responsible for statin-mediated eNOS upregulation. [Copyright &y& Elsevier]

Collagen plays a major role in arterial wall remodeling, aneurysm formation, and atherosclerotic cap stability. Smokers often have weakened arterial walls associating with aneurysm and thinned atherosclerotic plaque caps leading to rupture and acute coronary syndromes. We hypothesize that these detrimental effects on arterial wall by tobacco are partially mediated by disturbed collagen metabolism.We first investigated the effect of cigarette smoke extracts (CSE) on prolyl-4-hydroxylase (P4H) expression and collagen production in human aortic endothelial cells (HAECs) and human coronary artery smooth muscle cells (HCSMCs). After exposure to 0.01-U CSE for 24h, expression of P4Hα—a rate limiting subunit of P4H enzyme responsible for the formation of 4-hydroxyproline in mature functional collagen, was significantly down-regulated according to Western blotting and quantitative RT-PCR (HAEC p<0.01 and HCSMC p<0.001) when treated by CSE. The decreased P4Hα expression was corresponded with reduced cellular collagen levels (HAEC p<0.001 and HCSMC p<0.001). We also found that one of the cigarette components benzo(a)pyrene exerted similar effect as CSE, but not nicotine or acrolein. We further examined P4H expression in a few human atherosclerotic abdominal aortas. These in vivo data demonstrated that smokers had thinner atherosclerotic cap thickness and lower levels of P4Hα and collagen.Our study suggests that cigarette may interfere with one of the key enzymes in arterial wall collagen metabolism, which may be responsible for thin fibrous cap in atherosclerotic lesion, impaired arterial wall extensibility, and increased likelihood of aneurysm in smokers. [Copyright &y& Elsevier]

Genetic variation of fatty acid binding protein 2 (FABP2) may contribute to the high prevalence of obesity and Type II diabetes in Tonga. To explore this we assessed the frequency of the FABP2 Ala54Thr polymorphism, obesity, and Type II diabetes in Tongans and possible inter-relationships. We investigated 1022 Tongan subjects, 433 men and 589 women aged 15–85 years, to identify possible associations between the FABP2 Ala54Thr polymorphism, obesity, Type II diabetes, BMI, glucose tolerance and standard lipid variables. The prevalence of the polymorphism was compared with that reported for other ethnic populations (studies from: Japanese, Finnish, African American, Native Canadian and Inuit, Swedish, Guadeloupe Indians, European males, and Caucasian populations). We found that 84% of the Tongan men and 93% of the Tongan women were overweight or obese (BMI &ges; 25 kg/m2). The mean BMI ± SD was not significantly different among those who were and were not carrying the Thr allele (males: Ala/Ala 30.4 ± 5.4 and Thr carriers 29.8 ± 5.1; females: Ala/Ala 33.8 ± 6.4 and Thr carriers 33.6 ± 5.1). The genotype frequencies were 76.2% Ala/Ala, 22.8% Ala/Thr, and 1.0% Thr/Thr. The Alal/Ala frequency is higher than the prevalences reported for all populations studied. The Thr allele was significantly associated with lower total cholesterol and LDL cholesterol in both sexes and in women also with lower HDL cholesterol. We conclude that there is a high prevalence of the FABP2 Ala54Thr polymorphism in Tongans. The polymorphism may be involved in lipid metabolism as the Thr allele is associated with low total and LDL cholesterol levels in this population. [Copyright &y& Elsevier]

The major structural protein of the human polyomavirus BK (BKV), VP1, was expressed by using recombinant baculoviruses. A large amount of protein with a molecular mass of about 42 kDa was synthesized and identified by Western blotting. The protein was detected exclusively in the nuclei by immunofluorescent analysis and it was released into culture medium. The expressed BKV VP1 protein was self-assembled into virus-like particles (BK-VLPs) with two different sizes (50 and 26 nm in diameter), which migrated into four different bands in CsCl gradient with buoyant densities of 1.29, 1.30, 1.33, and 1.35 g/cm3. The immunological studies on the BK-VLPs suggested that they have similar antigenicity with those of authentic BKV particles. Cryoelectron microscopy and 3D image analysis further revealed that the larger BK-VLPs were composed of 72 capsomers which all were pentamers arranged in a T = 7 surface lattice. This system provides useful information for detailed studies of viral morphogenesis and the structural basis for the antigenicity of BKV. [Copyright &y& Elsevier]

With rapid improvements in industrialization and urbanization, antibiotics are now extensively used to prevent and treat human and animal diseases and husbandry and aquaculture. Some research has been conducted to assess the environmental distribution and risk level of antibiotics, but their distribution remains largely uncharacterized. Thus, this study investigated the distribution and abundance of 39 antibiotics belonging to five groups, and their associated risks in surface water around Luoma Lake in the north of Jiangsu province, China. Nineteen antibiotics were detected, at a detection frequency (DF) ranging from 2.27% to 100%. The total antibiotics (ΣABs) concentrations ranged from 34.91 to 825.93 ng/L, with a median concentration of 195.45 ng/L. Among these antibiotics, chlortetracycline (DF: 100%; median: 172.02 ng/L) was the dominant antibiotic, accounting for a median percentage of 91.0% of ΣABs concentrations. Spearman rank correlation method found a significant correlation between clindamycin (DF: 72.7%; median: 2.01 ng/L) and lincomycin (DF: 79.5%; median: 4.58 ng/L). The ecological risk quotient (RQ) values for two out of 44 sampling sites were higher than 1, indicating a high risk; 11.4% of the RQ values fell between 0.1 and 1, indicating a medium risk. Moreover, roxithromycin was found to be the dominant contributor to the ecological risk, accounting for a median of 79.7% of ΣABs. However, the total non-carcinogenic (<6.54 × 10−4) and carcinogenic risks (<1.64 × 10−7) of ΣABs were negligible at the detected concentrations. Image 1 • Concentrations and distributions of 39 antibiotics around Luoma Lake were revealed. • Chlortetracyclin was found to be the dominant antibiotic, with a median of 172.02 ng/L. • Ecological risks of antibiotics were at medium level (11.4%) or slightly high level (4.6%). • Non-carcinogenic and carcinogenic risks of antibiotics were low at the detected concentrations. [ABSTRACT FROM AUTHOR]

Service composition is a research hotspot with practical value. With the development of Web service, many Web services with the same functional attributes emerge. However, service composition optimization is still a big challenge since the complex and unstable composition environment. To solve this problem, we propose an adaptive service composition based on deep reinforcement learning, where recurrent neural network (RNN) is utilized for predicting the objective function, improving its expression and generalization ability, and effectively solving the shortcomings of traditional reinforcement learning in the face of large-scale or continuous state space problems. We leverage heuristic behavior selection strategy to divide the state set into hidden state and fully visible state. Effective simulation of hidden state space and fully visible state of the evaluation function can further improve the accuracy and efficiency of the combined results. We conduct comprehensive experiment and experimental results have shown the effectiveness of our method. [ABSTRACT FROM AUTHOR]