African swine fever virus (ASFV) is a contagious, rapidly spreading, transboundary animal disease and a major threat to pork production globally. Although plant-based feed has been identified as a ...potential route for virus introduction onto swine farms, little is known about the risks for ASFV transmission in feed. We aimed to determine the minimum and median infectious doses of the Georgia 2007 strain of ASFV through oral exposure during natural drinking and feeding behaviors. The minimum infectious dose of ASFV in liquid was 10
50% tissue culture infectious dose (TCID
), compared with 10
TCID
in feed. The median infectious dose was 10
TCID
for liquid and 10
TCID
for feed. Our findings demonstrate that ASFV Georgia 2007 can easily be transmitted orally, although higher doses are required for infection in plant-based feed. These data provide important information that can be incorporated into risk models for ASFV transmission.
Dengue virus (DENV) infection is a major cause of acute febrile illness in Indonesia. Diagnostic inaccuracy may occur due to its varied and non-specific presentation. Characterization of DENV ...epidemiology, clinical presentation, and virology will facilitate appropriate clinical management and public health policy.
A multicenter observational cohort study was conducted in Indonesia to assess causes of acute fever requiring hospitalization. Clinical information and specimens were collected at enrollment, 14-28 days, and 3 months from 1,486 children and adults. Total of 468 (31.9%) cases of DENV infection were confirmed by reference laboratory assays. Of these, 414 (88.5%) were accurately diagnosed and 54 had been misdiagnosed as another infection by sites. One hundred initially suspected dengue cases were finally classified as 'non-dengue'; other pathogens were identified in 58 of those cases. Mortality of DENV infection was low (0.6%). Prior DENV exposure was found in 92.3% of subjects >12 years. DENV circulated year-round in all cities, with higher incidence from January to March. DENV-3 and DENV-1 were the predominant serotypes. This study identified DENV-1 with TS119(C→T) substitution in the serotyping primer annealing site, leading to failure of serotype determination.
DENV is a common etiology of acute febrile illness requiring hospitalization in Indonesia. Diagnostic accuracy at clinical sites merits optimization since misdiagnosis of DENV infection and over-estimation of dengue can negatively impact management and outcomes. Mutation at the annealing site of the serotyping primer may confound diagnosis. Clinicians should consider following diagnostic algorithms that include DENV confirmatory testing. Policy-makers should prioritize development of laboratory capacity for diagnosis of DENV.
Nonalcoholic fatty liver disease (NAFLD) including nonalcoholic steatohepatitis (NASH) has reached epidemic proportions with no pharmacological therapy approved. Lower circulating glycine is ...consistently reported in patients with NAFLD, but the causes for reduced glycine, its role as a causative factor, and its therapeutic potential remain unclear. We performed transcriptomics in livers from humans and mice with NAFLD and found suppression of glycine biosynthetic genes, primarily alanine-glyoxylate aminotransferase 1 (
). Genetic (
mice) and dietary approaches to limit glycine availability resulted in exacerbated diet-induced hyperlipidemia and steatohepatitis, with suppressed mitochondrial/peroxisomal fatty acid β-oxidation (FAO) and enhanced inflammation as the underlying pathways. We explored glycine-based compounds with dual lipid/glucose-lowering properties as potential therapies for NAFLD and identified a tripeptide (Gly-Gly-L-Leu, DT-109) that improved body composition and lowered circulating glucose, lipids, transaminases, proinflammatory cytokines, and steatohepatitis in mice with established NASH induced by a high-fat, cholesterol, and fructose diet. We applied metagenomics, transcriptomics, and metabolomics to explore the underlying mechanisms. The bacterial genus
sensu stricto was markedly increased in mice with NASH and decreased after DT-109 treatment. DT-109 induced hepatic FAO pathways, lowered lipotoxicity, and stimulated de novo glutathione synthesis. In turn, inflammatory infiltration and hepatic fibrosis were attenuated via suppression of NF-κB target genes and TGFβ/SMAD signaling. Unlike its effects on the gut microbiome, DT-109 stimulated FAO and glutathione synthesis independent of NASH. In conclusion, impaired glycine metabolism may play a causative role in NAFLD. Glycine-based treatment attenuates experimental NAFLD by stimulating hepatic FAO and glutathione synthesis, thus warranting clinical evaluation.
Abstract Background Cardiac fibrosis (CF) is associated with increased ventricular stiffness and diastolic dysfunction and is an independent predictor of long-term clinical outcomes of patients with ...heart failure (HF). We previously showed that the matricellular CCN5 protein is cardioprotective via its ability to inhibit CF and preserve cardiac contractility. Objectives This study examined the role of CCN5 in human heart failure and tested whether CCN5 can reverse established CF in an experimental model of HF induced by pressure overload. Methods Human hearts were obtained from patients with end-stage heart failure. Extensive CF was induced by applying transverse aortic constriction for 8 weeks, which was followed by adeno-associated virus-mediated transfer of CCN5 to the heart. Eight weeks following gene transfer, cellular and molecular effects were examined. Results Expression of CCN5 was significantly decreased in failing hearts from patients with end-stage heart failure compared to nonfailing hearts. Trichrome staining and myofibroblast content measurements revealed that the established CF had been reversed by CCN5 gene transfer. Anti-CF effects of CCN5 were associated with inhibition of the transforming growth factor beta signaling pathway. CCN5 significantly inhibited endothelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation, which are 2 critical processes for CF progression, both in vivo and in vitro. In addition, CCN5 induced apoptosis in myofibroblasts, but not in cardiomyocytes or fibroblasts, both in vivo and in vitro. CCN5 provoked the intrinsic apoptotic pathway specifically in myofibroblasts, which may have been due the ability of CCN5 to inhibit the activity of NFκB, an antiapoptotic molecule. Conclusions CCN5 can reverse established CF by inhibiting the generation of and enhancing apoptosis of myofibroblasts in the myocardium. CCN5 may provide a novel platform for the development of targeted anti-CF therapies.
BACKGROUNDTo understand the features of a replicating vaccine that might drive potent and durable immune responses to transgene-encoded antigens, we tested a replication-competent adenovirus type 4 ...encoding influenza virus H5 HA (Ad4-H5-Vtn) administered as an oral capsule or via tonsillar swab or nasal spray.METHODSViral shedding from the nose, mouth, and rectum was measured by PCR and culturing. H5-specific IgG and IgA antibodies were measured by bead array binding assays. Serum antibodies were measured by a pseudovirus entry inhibition, microneutralization, and HA inhibition assays.RESULTSAd4-H5-Vtn DNA was shed from most upper respiratory tract-immunized (URT-immunized) volunteers for 2 to 4 weeks, but cultured from only 60% of participants, with a median duration of 1 day. Ad4-H5-Vtn vaccination induced increases in H5-specific CD4+ and CD8+ T cells in the peripheral blood as well as increases in IgG and IgA in nasal, cervical, and rectal secretions. URT immunizations induced high levels of serum neutralizing antibodies (NAbs) against H5 that remained stable out to week 26. The duration of viral shedding correlated with the magnitude of the NAb response at week 26. Adverse events (AEs) were mild, and peak NAb titers were associated with overall AE frequency and duration. Serum NAb titers could be boosted to very high levels 2 to 5 years after Ad4-H5-Vtn vaccination with recombinant H5 or inactivated split H5N1 vaccine.CONCLUSIONReplicating Ad4 delivered to the URT caused prolonged exposure to antigen, drove durable systemic and mucosal immunity, and proved to be a promising platform for the induction of immunity against viral surface glycoprotein targets.TRIAL REGISTRATIONClinicalTrials.gov NCT01443936 and NCT01806909.FUNDINGIntramural and Extramural Research Programs of the NIAID, NIH (U19 AI109946) and the Centers of Excellence for Influenza Research and Surveillance (CEIRS), NIAID, NIH (contract HHSN272201400008C).
Metformin, an oral antihyperglycemic, is increasingly being prescribed to pregnant women with gestational diabetes. Metformin is a hydrophilic cation and relies on organic cation transporters to move ...across cell membranes. We previously demonstrated that human and mouse placentas predominantly express organic cation transporter 3 (OCT3), but the impact of this transporter on maternal and fetal disposition of metformin is unknown. Using immunofluorescence colocalization studies in term human placenta, we showed that OCT3 is localized to the basal (fetal-facing) membrane of syncytiotrophoblast cells with no expression on the apical (maternal-facing) membrane. OCT3 positive staining was also observed in fetal capillaries. To determine the in vivo role of OCT3 in maternal and fetal disposition of metformin, we determined metformin maternal pharmacokinetics and overall fetal exposure in wild-type and
-null pregnant mice. After oral dosing of
Cmetformin at gestational day 19, the systemic drug exposure (AUC
) in maternal plasma was slightly reduced by ∼16% in the
pregnant mice. In contrast, overall fetal AUC
was reduced by 47% in the
pregnant mice. Consistent with our previous findings in nonpregnant mice, metformin tissue distribution was respectively reduced by 70% and 52% in the salivary glands and heart in
pregnant mice. Our in vivo data in mice clearly demonstrated a significant role of Oct3 in facilitating metformin fetal distribution and exposure during pregnancy. Modulation of placental OCT3 expression or activity by gestational age, genetic polymorphism, or pharmacological inhibitors may alter fetal exposure to metformin or other drugs transported by OCT3.
A model wetland with Salix was established to investigate the effects of CuO nanoparticles (NPs; the equivalent amount of Cu at 0, 100 and 500 mg/kg) on plant, soil enzyme activity and microbial ...community. Ionic Cu (100, 500 mg/kg) and bulk-sized CuO particles (BPs, 500 mg/kg) were included as controls. The results suggested the CuO NPs at 500 mg/kg and ionic Cu treatments inhibited the plant growth, while CuO NPs at 100 mg/kg and CuO BPs at 500 mg/kg played a facilitating role. CuO NPs significantly decreased the activities of peroxidase and polyphenol oxidase, while ionic Cu treatments increased peroxidase activity, BPs and ionic Cu (500 mg/kg) increased the polyphenol oxidase activity. Bacterial community richness and diversity were reduced in all Cu treatments; however, CuO NPs and BPs at 500 mg/kg significantly increased the richness and diversity of fungal community.Soil microbial community was significantly altered by Cu types and dose. In comparison with ionic Cu and CuO BPs, CuO NPs uniquely enriched the microbial community and the fungal families.Overall, it demonstrate that both particle size and dose regulate the impact of CuO on wetland ecology, which deepens our understanding on the ecological risks of CuO NPs in freshwater forested wetland.
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•The process of natural release of CuO NPs into wetland environment was simulated.•The morphology of Cu directly affects its distribution pattern in water, soil and plant.•CuO NPs at 100 mg/kg promoted the growth of Salix, but inhibited at 500 mg/kg.•Soil microbial community was significantly altered by Cu types and dose.•Both size and dose regulate the impact of CuO on soil microbiota and plant growth.
Movies vary widely in appeal, star power, cost, and other elements, and therefore, each might be expected to charge a different price. Multiplexes, however, typically charge the same price for all ...movies, except for such premium formats as 3D, a choice that has puzzled managers and researchers. Because of data limitations, minimal empirical work has directly addressed this issue. In Hong Kong, however, prices vary both within and across multiplexes. Using daily ticket prices and attendance by theater and movie, the authors empirically examine the potential gains from differentiated movie-specific pricing as well as the increasingly common two-tier (2D/3D) uniform pricing, as compared with a full uniform pricing strategy in which a theater charges the same price for all its movies. Their results show that differential pricing leads to higher profits than the two-tier uniform pricing practice, but that the improvement is limited. In contrast, the gains are substantial when compared with the full uniform pricing strategy, suggesting that only minimal differentiation (2D/3D) may obtain most of the gains available from fully differentiated prices.
Objectives The coronavirus disease 2019 pandemic has affected countries around the world since 2020, and an increasing number of people are being infected. The purpose of this research was to use big ...data and artificial intelligence technology to find key factors associated with the coronavirus disease 2019 infection. The results can be used as a reference for disease prevention in practice. Methods This study obtained data from the "Imperial College London YouGov Covid-19 Behaviour Tracker Open Data Hub", covering a total of 291,780 questionnaire results from 28 countries (April 1~August 31, 2020). Data included basic characteristics, lifestyle habits, disease history, and symptoms of each subject. Four types of machine learning classification models were used, including logistic regression, random forest, support vector machine, and artificial neural network, to build prediction modules. The performance of each module is presented as the area under the receiver operating characteristics curve. Then, this study further processed important factors selected by each module to obtain an overall ranking of determinants. Results This study found that the area under the receiver operating characteristics curve of the prediction modules established by the four machine learning methods were all >0.95, and the RF had the highest performance (area under the receiver operating characteristics curve is 0.988). Top ten factors associated with the coronavirus disease 2019 infection were identified in order of importance: whether the family had been tested, having no symptoms, loss of smell, loss of taste, a history of epilepsy, acquired immune deficiency syndrome, cystic fibrosis, sleeping alone, country, and the number of times leaving home in a day. Conclusions This study used big data from 28 countries and artificial intelligence methods to determine the predictors of the coronavirus disease 2019 infection. The findings provide important insights for the coronavirus disease 2019 infection prevention strategies.
Plasmacytoid dendritic cells (PDCs) and their production of interferon-alpha (IFN-α) are believed to play an important role in human immunodeficiency virus, type I (HIV-1) pathogenesis. PDCs produce ...IFN-α and other proinflammatory cytokines through stimulation of Toll-like receptor 7 (TLR7) and TLR9 present in endosomal compartments. TLR7 recognizes single-stranded viral RNA, while TLR9 recognizes unmethylated DNA. In this study, we examined the mechanisms that may underlie variations in IFN-α production in response to HIV, and the impact of these variations on HIV pathogenesis. In four distinct cohorts, we examined PDC production of IFN-α upon stimulation with inactivated HIV-1 particles and unmethylated DNA. The signaling cascade of TLR7 bifurcates at the myeloid differentiation protein 88 (MyD88) adaptor protein to induce expression of either IFN-α or TNF-α. To determine whether variations in IFN-α production are modulated at the level of the receptor complex or downstream of it, we correlated production of IFN-α and TNF-α following stimulation of TLR7 or TLR9 receptors. Flow cytometry detection of intracellular cytokines showed strong, direct correlations between IFN-α and TNF-α expression in all four cohorts, suggesting that variations in IFN-α production are not due to variations downstream of the receptor complex. We then investigated the events upstream of TLR binding by using lipid-like vesicles to deliver TLR ligands directly to the TLR receptors, bypassing the need for CD4 binding and endocytosis. Similar tight correlations were found in IFN-α and TNF-α production in response to the TLR ligands. Taken together, these results strongly suggest that differences in IFN-α production depend on the regulatory processes at the level of the TLR7 receptor complex. Additionally, we found no association between IFN-α production before HIV infection and disease progression.
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