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1.
Vet Pathol ; 61(4): 609-620, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38323378

RESUMO

Between September and November 2021, 5 snow leopards (Panthera uncia) and 1 lion (Panthera leo) were naturally infected with severe acute respiratory coronavirus 2 (SARS-CoV-2) and developed progressive respiratory disease that resulted in death. Severe acute respiratory syndrome coronavirus 2 sequencing identified the delta variant in all cases sequenced, which was the predominant human variant at that time. The time between initial clinical signs and death ranged from 3 to 45 days. Gross lesions in all 6 cats included nasal turbinate hyperemia with purulent discharge and marked pulmonary edema. Ulcerative tracheitis and bronchitis were noted in 4 cases. Histologically, there was necrotizing and ulcerative rhinotracheitis and bronchitis with fibrinocellular exudates and fibrinosuppurative to pyogranulomatous bronchopneumonia. The 4 cats that survived longer than 8 days had fungal abscesses. Concurrent bacteria were noted in 4 cases, including those with more acute disease courses. Severe acute respiratory syndrome coronavirus 2 was detected by in situ hybridization using probes against SARS-CoV-2 spike and nucleocapsid genes and by immunohistochemistry. Viral nucleic acid and protein were variably localized to mucosal and glandular epithelial cells, pneumocytes, macrophages, and fibrinocellular debris. Based on established criteria, SARS-CoV-2 was considered a contributing cause of death in all 6 cats. While mild clinical infections are more common, these findings suggest that some SARS-CoV-2 variants may cause more severe disease and that snow leopards may be more severely affected than other felids.


Assuntos
COVID-19 , SARS-CoV-2 , Animais , COVID-19/veterinária , COVID-19/virologia , COVID-19/patologia , COVID-19/mortalidade , Feminino , Masculino , Leões/virologia , Panthera/virologia , Pulmão/patologia , Pulmão/virologia , Gatos , Felidae/virologia , Doenças do Gato/virologia , Doenças do Gato/patologia
2.
BMC Public Health ; 24(1): 1071, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38632605

RESUMO

BACKGROUND: Inter-leg systolic blood pressure difference (ILSBPD) has emerged as a novel cardiovascular risk factor. This study aims to investigate the predictive value of ILSBPD on all-cause and cardiovascular mortality in general population. METHODS: We combined three cycles (1999-2004) of the National Health and Nutrition Examination Survey (NHANES) data. Levels of ILSBPD were calculated and divided into four groups based on three cut-off values of 5, 10 and 15mmHg. Time-to-event curves were estimated with the use of the Kaplan-Meier method, and two multivariable Cox proportional hazards regression models were conducted to assess the hazard ratios (HRs) and 95% confidence intervals (CIs) of all-cause and cardiovascular mortality associated with ILSBPD. RESULTS: A total of 6 842 subjects were included, with the mean (SD) age of 59.5 (12.8) years. By December 31, 2019, 2 544 and 648 participants were identified all-cause and cardiovascular mortality respectively during a median follow-up of 16.6 years. Time-to-event analyses suggested that higher ILSBPD was associated with increased all-cause and cardiovascular mortality (logrank, p < 0.001). Every 5mmHg increment of ILSBPD brings about 5% and 7% increased risk of all-cause and cardiovascular mortality, and individuals with an ILSBPD ≥ 15mmHg were significantly associated with higher incidence of all-cause mortality (HR 1.43, 95%CI 1.18-1.52, p < 0.001) and cardiovascular mortality (HR 1.73, 95%CI 1.36-2.20, p < 0.001) when multiple confounding factors were adjusted. Subgroup and sensitivity analysis confirmed the relationship. CONCLUSIONS: Our findings suggest that the increment of ILSBPD was significantly associated with higher risk of all-cause and cardiovascular mortality in general population.


Assuntos
Doenças Cardiovasculares , Humanos , Pessoa de Meia-Idade , Pressão Sanguínea/fisiologia , Inquéritos Nutricionais , Doenças Cardiovasculares/epidemiologia , Perna (Membro) , Fatores de Risco
3.
Mol Cell ; 60(3): 475-86, 2015 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-26593721

RESUMO

The regulation of protein synthesis contributes to gene expression in both normal physiology and disease, yet kinetic investigations of the human translation mechanism are currently lacking. Using single-molecule fluorescence imaging methods, we have quantified the nature and timing of structural processes in human ribosomes during single-turnover and processive translation reactions. These measurements reveal that functional complexes exhibit dynamic behaviors and thermodynamic stabilities distinct from those observed for bacterial systems. Structurally defined sub-states of pre- and post-translocation complexes were sensitive to specific inhibitors of the eukaryotic ribosome, demonstrating the utility of this platform to probe drug mechanism. The application of three-color single-molecule fluorescence resonance energy transfer (smFRET) methods further revealed a long-distance allosteric coupling between distal tRNA binding sites within ribosomes bearing three tRNAs, which contributed to the rate of processive translation.


Assuntos
Biossíntese de Proteínas , RNA de Transferência/química , Ribossomos/química , Regulação Alostérica , Transferência Ressonante de Energia de Fluorescência , Humanos , RNA de Transferência/metabolismo , Ribossomos/metabolismo
4.
Anim Biotechnol ; 34(9): 4783-4792, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37022008

RESUMO

The proliferation and myogenic differentiation of muscle stem cells (MuSCs) are important factors affecting muscle development and beef quality. There is increasing evidence that circRNAs can regulate myogenesis. We found a novel circRNA, named circRRAS2 that is significantly upregulated in the differentiation phase of bovine MuSCs. Here, we aimed to determine its roles in the proliferation and myogenic differentiation of these cells. The results showed that circRRAS2 was expressed in several bovine tissues. CircRRAS2 inhibited MuSCs proliferation and promoted myoblast differentiation. In addition, chromatin isolation by using RNA purification and mass spectrometry in differentiated muscle cells identified 52 RNA-binding proteins that could potentially bind to circRRAS2, in order to regulate their differentiation. The results suggest that circRRAS2 could be a specific regulator of myogenesis in bovine muscle.HighlightsCircRRAS2 expression is higher in DM cells than in GM cells.CircRRAS2 could significantly inhibit the proliferation and apoptosis of bovine MuSCs.CircRRAS2 promotes the differentiation of bovine MuSCs into myotubes.CircRRAS2 may exert regulatory effects through multiple RNA binding proteins.


Assuntos
Células Satélites de Músculo Esquelético , Bovinos , Animais , Diferenciação Celular/genética , Células Cultivadas , Linhagem Celular , Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , Proliferação de Células/genética
5.
J Environ Manage ; 344: 118478, 2023 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-37393876

RESUMO

The implementation of Personal Carbon Trading (PCT) holds promise in facilitating a noteworthy contribution towards the attainment of emissions reduction predicated on consumption patterns and consequently motivating lifestyle modifications. As individual consumption behaviors usually lead to continuous changes in carbon emissions, it is crucial to rethink PCT from a systematic perspective. This review employed a bibliometric analysis of 1423 papers related to PCT, highlighting the key themes of carbon emissions from energy consumption, climate change, and public opinion on policies in the context of PCT. Most of the existing PCT researches focus on theoretical assumptions and public attitudes, while the quantification of carbon emissions and simulation of PCT require further investigation. Furthermore, the concept of Tan Pu Hui is seldom addressed in PCT studies and case analyses. Moreover, there are limited PCT schemes worldwide that can be directly implemented in practice, leading to a scarcity of large-scale, high-participation case studies. To address these gaps, this review proposes a framework to clarify how PCT can stimulate individual emission reductions on the consumption side, comprising two phases, from motivation to behavior and behavior to target. Future endeavors should prioritize the enhancement of the systematic study of the theoretical foundation of PCT, encompassing carbon emissions accounting and policy design, the incorporation of cutting-edge technology, and the reinforcement of integrated policy practice. This review serves as a valuable reference for future research endeavors and policymaking efforts.


Assuntos
Carbono , Formulação de Políticas , Mudança Climática , Simulação por Computador , Dióxido de Carbono/análise , China
6.
J Zoo Wildl Med ; 54(3): 607-616, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37817628

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in nondomestic felids have been documented in North America, South America, Africa, Europe, and Asia. Between March 2020 and February 2021, at nine institutions across three continents, infection was confirmed in 16 tigers (Panthera tigris), 14 lions (Panthera leo), three snow leopards (Panthera uncia), one cougar (Puma concolor), and one Amur leopard cat (Prionailurus bengalensis euptilurus) ranging from 2 to 21 yr old (average, 10 yr). Infection was suspected in an additional 12 tigers, 4 lions, and 9 cougars. Clinical signs (in order of most to least common) included coughing, ocular and/or nasal discharge, wheezing, sneezing, decreased appetite, lethargy, diarrhea, and vomiting. Most felids recovered uneventfully, but one geriatric tiger with comorbidities developed severe dyspnea and neurologic signs necessitating euthanasia. Clinical signs lasted 1-19 d (average, 8 d); one tiger was asymptomatic. Infection was confirmed by various methods, including antigen tests and/or polymerase chain reaction (PCR) of nasal or oral swabs, tracheal wash, and feces, or virus isolation from feces or tracheal wash. Infection status and resolution were determined by testing nasal swabs from awake animals, fecal PCR, and observation of clinical signs. Shedding of fecal viral RNA was significantly longer than duration of clinical signs. Postinfection seropositivity was confirmed by four institutions including 11 felids (5 lions, 6 tigers). In most instances, asymptomatic or presymptomatic keepers were the presumed or confirmed source of infection, although in some instances the infection source remains uncertain. Almost all infections occurred despite using cloth facemasks and disposable gloves when in proximity to the felids and during food preparation. Although transmission may have occurred during momentary lapses in personal protective equipment compliance, it seems probable that cloth masks are insufficient at preventing transmission of SARS-CoV-2 from humans to nondomestic felids. Surgical or higher grade masks may be warranted when working with nondomestic felids.


Assuntos
COVID-19 , Felidae , Leões , Panthera , Tigres , Humanos , Animais , COVID-19/veterinária , Estudos Retrospectivos , SARS-CoV-2
7.
Postepy Dermatol Alergol ; 40(5): 670-678, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38028411

RESUMO

Introduction: Dupilumab is approved for a variety of type 2 inflammatory diseases. Changes in chemokine levels during treatment require further analysis. Aim: We evaluated changes in eotaxin-3 and PARC levels after dupilumab treatment through a meta-analysis, aiming to provide more comprehensive results. Material and methods: Databases were searched to select eligible publications. The study quality was assessed after inclusion. The standardized mean difference (SMD) was used for evaluation. Results: Four studies were included. Eotaxin-3 levels were not seen significantly decreased at weeks 1 and 12, with SMD = -0.39 (95% CI: -1.78, 0.99) and -2.60 (95% CI: -5.77, 0.57), respectively (p > 0.05). Eotaxin-3 levels decreased significantly at weeks 2, 4, 8, 16, 24, 36, and 52, with SMD = -0.94 (95% CI: -1.61, -0.27); -1.17 (95% CI: -1.49, -0.84); -1.20 (95% CI: -1.52, -0.88); -1.31 (95% CI: -1.83, -0.79); -4.57 (95% CI: -6.90, -2.33); -5.28 (95% CI: -5.52, -5.04); and -4.03 (95% CI: -4.22, -3.85) (p < 0.05), respectively. PARC levels decreased significantly at weeks 4, 8, 12, and 16, with SMD = -1.08 (95% CI: -1.59, -0.58); -1.17 (95% CI: -1.68, -0.66); -1.11 (95% CI: -1.61, -0.60); and -1.15 (95% CI: -1.66, -0.64) (p < 0.05), respectively. Conclusions: Eotaxin-3 and PARC levels can be significantly reduced in patients treated with dupilumab.

8.
Liver Transpl ; 28(11): 1756-1765, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35665591

RESUMO

The burden of early hospitalization (within 6 months) following simultaneous liver-kidney transplant (SLKT) is not known. We examined risk factors associated with early hospitalization after SLKT and their impact on patient mortality conditional on 6-month survival. We used data from the US Multicenter SLKT Consortium cohort study of all adult SLKT recipients between 2002 and 2017 who were discharged alive following SLKT. We used Poisson regression to model rates of early hospitalizations after SLKT. Cox regression was used to identify risk factors associated with mortality conditional on survival at 6 months after SLKT. Median age (N = 549) was 57.7 years (interquartile range [IQR], 50.6-63.9) with 63% males and 76% Whites; 33% had hepatitis C virus, 20% had non-alcohol-associated fatty liver disease, 23% alcohol-associated liver disease, and 24% other etiologies. Median body mass index (BMI) and Model for End-Stage Liver Disease-sodium scores were 27.2 kg/m2 (IQR, 23.6-32.2 kg/m2 ) and 28 (IQR, 23-34), respectively. Two-thirds of the cohort had at least one hospitalization within the first 6 months of SLKT. Age, race, hospitalization at SLKT, diabetes mellitus, BMI, and discharge to subacute rehabilitation (SAR) facility after SLKT were independently associated with a high incidence rate ratio of early hospitalization. Number of hospitalizations within the first 6 months did not affect conditional survival. Early hospitalizations after SLKT were very common but did not affect conditional survival. Although most of the risk factors for early hospitalization were nonmodifiable, discharge to SAR after initial SLKT was associated with a significantly higher incidence rate of early hospitalization. Efforts and resources should be focused on identifying SLKT recipients at high risk for early hospitalization to optimize their predischarge care, discharge planning, and long-term follow-up.


Assuntos
Doença Hepática Terminal , Transplante de Rim , Transplante de Fígado , Adulto , Estudos de Coortes , Doença Hepática Terminal/complicações , Feminino , Sobrevivência de Enxerto , Hospitalização , Humanos , Transplante de Rim/métodos , Transplante de Fígado/métodos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Índice de Gravidade de Doença , Sódio , Resultado do Tratamento
9.
Appl Environ Microbiol ; 88(7): e0228921, 2022 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-35285246

RESUMO

Monitoring the prevalence of SARS-CoV-2 variants is necessary to make informed public health decisions during the COVID-19 pandemic. PCR assays have received global attention, facilitating a rapid understanding of variant dynamics because they are more accessible and scalable than genome sequencing. However, as PCR assays target only a few mutations, their accuracy could be reduced when these mutations are not exclusive to the target variants. Here we introduce PRIMES, an algorithm that evaluates the sensitivity and specificity of SARS-CoV-2 variant-specific PCR assays across different geographical regions by incorporating sequences deposited in the GISAID database. Using PRIMES, we determined that the accuracy of several PCR assays decreased when applied beyond the geographic scope of the study in which the assays were developed. Subsequently, we used this tool to design Alpha and Delta variant-specific PCR assays for samples from Illinois, USA. In silico analysis using PRIMES determined the sensitivity/specificity to be 0.99/0.99 for the Alpha variant-specific PCR assay and 0.98/1.00 for the Delta variant-specific PCR assay in Illinois, respectively. We applied these two variant-specific PCR assays to six local sewage samples and determined the dominant SARS-CoV-2 variant of either the wild type, the Alpha variant, or the Delta variant. Using next-generation sequencing (NGS) of the spike (S) gene amplicons of the Delta variant-dominant samples, we found six mutations exclusive to the Delta variant (S:T19R, S:Δ156/157, S:L452R, S:T478K, S:P681R, and S:D950N). The consistency between the variant-specific PCR assays and the NGS results supports the applicability of PRIMES. IMPORTANCE Monitoring the introduction and prevalence of variants of concern (VOCs) and variants of interest (VOIs) in a community can help the local authorities make informed public health decisions. PCR assays can be designed to keep track of SARS-CoV-2 variants by measuring unique mutation markers that are exclusive to the target variants. However, the mutation markers may not be exclusive to the target variants because of regional and temporal differences in variant dynamics. We introduce PRIMES, an algorithm that enables the design of reliable PCR assays for variant detection. Because PCR is more accessible, scalable, and robust for sewage samples than sequencing technology, our findings will contribute to improving global SARS-CoV-2 variant surveillance.


Assuntos
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , COVID-19/epidemiologia , Humanos , Mutação , Pandemias , Reação em Cadeia da Polimerase , SARS-CoV-2/genética , Esgotos
10.
J Infect Dis ; 224(6): 976-982, 2021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34191025

RESUMO

BACKGROUND: Serial screening is critical for restricting spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by facilitating timely identification of infected individuals to interrupt transmission. Variation in sensitivity of different diagnostic tests at different stages of infection has not been well documented. METHODS: In a longitudinal study of 43 adults newly infected with SARS-CoV-2, all provided daily saliva and nasal swabs for quantitative reverse transcription polymerase chain reaction (RT-qPCR), Quidel SARS Sofia antigen fluorescent immunoassay (FIA), and live virus culture. RESULTS: Both RT-qPCR and Quidel SARS Sofia antigen FIA peaked in sensitivity during the period in which live virus was detected in nasal swabs, but sensitivity of RT-qPCR tests rose more rapidly prior to this period. We also found that serial testing multiple times per week increases the sensitivity of antigen tests. CONCLUSIONS: RT-qPCR tests are more effective than antigen tests at identifying infected individuals prior to or early during the infectious period and thus for minimizing forward transmission (given timely results reporting). All tests showed >98% sensitivity for identifying infected individuals if used at least every 3 days. Daily screening using antigen tests can achieve approximately 90% sensitivity for identifying infected individuals while they are viral culture positive.


Assuntos
Teste para COVID-19 , COVID-19/diagnóstico , Testes Diagnósticos de Rotina , SARS-CoV-2/isolamento & purificação , Adulto , Idoso , Animais , Antígenos Virais/análise , Chlorocebus aethiops , Feminino , Humanos , Estudos Longitudinais , Masculino , Programas de Rastreamento , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Saliva , Sensibilidade e Especificidade , Células Vero , Adulto Jovem
11.
J Virol ; 94(12)2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32295907

RESUMO

Mumps virus (MuV) caused the most viral meningitis before mass immunization. Unfortunately, MuV has reemerged in the United States in the past several years. MuV is a member of the genus Rubulavirus, in the family Paramyxoviridae, and has a nonsegmented negative-strand RNA genome. The viral RNA-dependent RNA polymerase (vRdRp) of MuV consists of the large protein (L) and the phosphoprotein (P), while the nucleocapsid protein (NP) encapsulates the viral RNA genome. These proteins make up the replication and transcription machinery of MuV. The P protein is phosphorylated by host kinases, and its phosphorylation is important for its function. In this study, we performed a large-scale small interfering RNA (siRNA) screen targeting host kinases that regulated MuV replication. The human kinase ribosomal protein S6 kinase beta-1 (RPS6KB1) was shown to play a role in MuV replication and transcription. We have validated the role of RPS6KB1 in regulating MuV using siRNA knockdown, an inhibitor, and RPS6KB1 knockout cells. We found that MuV grows better in cells lacking RPS6KB1, indicating that it downregulates viral growth. Furthermore, we detected an interaction between the MuV P protein and RPS6KB1, suggesting that RPS6KB1 directly regulates MuV replication and transcription.IMPORTANCE Mumps virus is an important human pathogen. In recent years, MuV has reemerged in the United State, with outbreaks occurring in young adults who have been vaccinated. Our work provides insight into a previously unknown mumps virus-host interaction. RPS6KB1 negatively regulates MuV replication, likely through its interaction with the P protein. Understanding virus-host interactions can lead to novel antiviral drugs and enhanced vaccine production.


Assuntos
Genoma Viral , Vírus da Caxumba/genética , Proteínas do Nucleocapsídeo/genética , Fosfoproteínas/genética , RNA Polimerase Dependente de RNA/genética , Proteínas Quinases S6 Ribossômicas 70-kDa/genética , Proteínas Virais/genética , Animais , Chlorocebus aethiops , Regulação da Expressão Gênica , Células HEK293 , Células HeLa , Interações Hospedeiro-Patógeno/genética , Humanos , Vírus da Caxumba/metabolismo , Proteínas do Nucleocapsídeo/metabolismo , Fosfoproteínas/metabolismo , Fosforilação , Ligação Proteica , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , RNA Polimerase Dependente de RNA/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/antagonistas & inibidores , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Transdução de Sinais , Células Vero , Proteínas Virais/metabolismo , Replicação Viral
12.
Stem Cells ; 38(1): 134-145, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31644832

RESUMO

The purpose of this study was to investigate the effects of secreted protein acidic and rich in cysteine (SPARC) on the maintenance of limbal epithelial stem cell (LESC) stemness and restoration of ocular surface. To determine the suitable concentration of SPARC for LESC culture, the marker expression, mitogenic effect, and holoclone-forming capacity of LESCs treated with different concentrations of SPARC were analyzed. To investigate the mechanism of SPARC's action on the preservation of LESCs stemness, the phosphorylation of related signaling pathways was evaluated by Western blotting. A corneal wound model was established to verify the function of SPARC in ocular surface repair. Consecutive subculturing, colony-forming efficiency, immunofluorescence, and 5-ethynyl-2-deoxyuridine incorporation assays indicated that 1 µg/mL SPARC was a suitable concentration to stimulate LESC proliferation and preserve their proliferative potential. Compared with a control group, 1 µg/mL SPARC effectively increased the expression of ABCG-2, Bmi-1, and Ki67, while decreasing that of CK3/12. The mitogenic effect of SPARC on LESCs was found to be mediated by the phosphorylation of c-Jun N-terminal kinase (JNK) and p38-MAPK signaling pathways, whereas the inhibitors of JNK and p38 MAPK reduced the marker expression and mitogenic capacity of LESCs. In a corneal injury model, SPARC facilitated corneal epithelial wound healing and promoted the proliferation of p63α-positive cells both in the limbus and in the epithelial healing front. SPARC promotes proliferation while suppressing spontaneous differentiation of LESCs through JNK and p38-MAPK signaling pathways, suggesting that SPARC is a promising factor for the improvement of LESCs culture in vitro and in vivo.


Assuntos
Células Epiteliais/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Limbo da Córnea/metabolismo , Osteonectina/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Células 3T3 , Animais , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Células Epiteliais/citologia , Humanos , Sistema de Sinalização das MAP Quinases , Camundongos , Coelhos
13.
Exp Eye Res ; 209: 108644, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34081998

RESUMO

Fungal keratitis constitutes a serious vision-threatening disease. Toll-like receptors (TLRs) comprise key mediators of innate immunity triggered by Aspergillus fumigatus (AF) in the cornea, but the messenger between innate and adaptive immunity remained unknown. Thymic stromal lymphopoietin (TSLP) represents a critical factor of adaptive immunity. Here we investigated the expression of TSLP in corneal epithelial and stromal cells challenged by AF and its relationship with TLRs. We stimulated corneal cells with TLR ligands zymosan or lipopolysaccharide (LPS), human recombinant TSLP, or AF hyphae for various periods, with or without prior TLR2, TLR4, or TSLP inhibition. TLR2, TLR4, TSLP, IL-8, and TNF-α release and expression were measured via enzyme-linked immunosorbent analysis, quantitative polymerase chain reaction, or western blot. Corneal cell stimulation with zymosan or LPS induced up-regulated TSLP expression. Enhanced TSLP expression was associated with AF treatment in human corneal cells; TLR2 or TLR4 inhibition impaired the AF-induced TSLP levels. Human recombinant TSLP augmented TLR2 and TLR4 expression; RNA interference of TSLP attenuated TLR, IL-8, and TNF-α expression stimulated by AF hyphae. These findings indicated that TSLP participates in the immune response of corneal cells triggered by AF, which is closely related to TLR function, and the innate immunity mediated by TLRs could be enhanced by TSLP. Innate immunity may therefore transmit inflammatory signals to adaptive immunity through activation of TSLP; in turn, adaptive immunity likely exerts certain regulatory effects on innate immunity via TSLP. That is, TSLP could interact with innate immunity mediated by TLR2 and TLR4 in human corneal cells challenged by AF and thus may serve as a messenger between the innate and adaptive immune responses in AF keratitis.


Assuntos
Aspergilose/genética , Aspergillus fumigatus/imunologia , Citocinas/genética , Regulação da Expressão Gênica , Ceratite/genética , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética , Aspergilose/microbiologia , Aspergilose/patologia , Células Cultivadas , Citocinas/biossíntese , Ensaio de Imunoadsorção Enzimática , Epitélio Corneano/metabolismo , Epitélio Corneano/microbiologia , Epitélio Corneano/patologia , Infecções Oculares Fúngicas/microbiologia , Humanos , Imunidade Inata , Ceratite/metabolismo , Ceratite/patologia , RNA/genética , Células Estromais , Receptor 2 Toll-Like/biossíntese , Receptor 4 Toll-Like/biossíntese , Linfopoietina do Estroma do Timo
14.
Exp Eye Res ; 202: 108366, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33227296

RESUMO

Fungal keratitis is a serious corneal infection, which can lead to significant visual impairment and blindness. The cGAS-STING signaling pathway has emerged as a key player in innate immunity by sensing of invading pathogens. However, the role of the cGAS-STING pathway in Aspergillus fumigatus (A. fumigatus) keratitis is still unknown. In this study, we showed that the cGAS-STING signaling pathway was activated in human corneal epithelial cells (HCECs) and in mouse corneas infected with A. fumigatus. Knockdown of cGAS reduced A. fumigatus-induced production of pro-inflammatory cytokines, including TNF-α, IL-1ß, IL-6, and IFN-ß. However, reconstruction of cGAS activity restored the inflammatory response in HCECs infected with A. fumigatus. A specific cGAS inhibitor, RU.521, could also significantly inhibit A. fumigatus-induced inflammatory cytokine expression. In addition, we found that cGAS was indispensable for the autophagy flux evoked by A. fumigatus infection. Moreover, inhibition of cGAS using siRNA or RU.521 alleviated the severity of A. fumigatus keratitis in the mouse cornea. Therefore, the cGAS-STING signaling pathway contributes to the progression of A. fumigatus keratitis and targeting this pathway may provide therapeutic potential.


Assuntos
Aspergilose/genética , Infecções Oculares Fúngicas/genética , Regulação da Expressão Gênica , Imunidade Inata , Ceratite/genética , Proteínas de Membrana/genética , Nucleotidiltransferases/genética , Animais , Aspergilose/diagnóstico , Aspergilose/metabolismo , Aspergillus fumigatus/imunologia , Autofagia , Modelos Animais de Doenças , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/metabolismo , Ceratite/diagnóstico , Ceratite/metabolismo , Masculino , Proteínas de Membrana/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Nucleotidiltransferases/biossíntese , RNA/genética , RNA/metabolismo , Transdução de Sinais
15.
BMC Ophthalmol ; 21(1): 302, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34399712

RESUMO

BACKGROUND: Retinitis pigmentosa (RP) is a rare, progressive, and hereditary disorder that leads to the progressive loss of vision and visual field, and in some cases blindness. The specific relationship between RP and glaucoma has been debated for decades. METHODS: In this study, we examined a Han RP family with concomitant angle-closure glaucoma (ACG), performed an inductive analysis of their clinical features and assistant results, and applied whole-exome sequencing (WES) technology for a molecular diagnosis. RESULTS: A novel transversion mutation (c.626 T > A) was identified in the peripherin-2 (PRPH2) gene in the proband, resulting in the substitution of Valine to aspartic acid in codon 209. A full ophthalmic examination showed that the proband with the c.626 T > A mutation had a typical RP manifestation, with close angles; however, the proband's elder brother, who lacked the novel mutation, had a normal fundus and open angles. CONCLUSION: Our results extend the genetic mutation spectrum of PRPH2 in RP, and provide evidence to support a genetic correlation between RP and ACG.


Assuntos
Glaucoma de Ângulo Fechado , Retinose Pigmentar , Idoso , China , Glaucoma de Ângulo Fechado/genética , Humanos , Masculino , Mutação , Linhagem , Periferinas , Retinose Pigmentar/genética
16.
Chem Biodivers ; 18(1): e2000797, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33245200

RESUMO

The microbial production of dextranase using cheap carbon sources is beneficial to solve the economic loss caused by the accumulation of dextran in syrup. A food-grade microbial cell factory was constructed by introducing the dextranase encoding gene DEX from Chaetomium gracile to the chromosome of Bacillus subtilis, and the antibiotic resistance marker gene was subsequently deleted via the Cre/loxP strategy. The dual-promoter system with a sequentially arranged constitutive P43 promoter resulted in an 85 % increase in DEX expression. Under the optimal fermentation conditions of 10 g/L maltose, 15 g/L casein, 1 g/L Na2 HPO4 , 1 g/L FeSO4 and 8 g/L NaCl, DEX activity was increased from 2.625 to 64.34 U/mL. Recombinant DEX was purified 5.98-fold with a recovery ratio of 26.67 % and specific activity of 3935.02 U/mg. Enzyme activity was optimal at 55 °C and pH 5.0 and remained 80.34 % and 71.36 % of the initial activity at 55 °C and pH 4.0 after 60 min, respectively. The enzyme possessed high activity in the presence of Co2+ , while Ag+ showed the strongest inhibition ability. The optimal substrate was 20 g/L dextran T-2000. The findings could facilitate the low-cost, large-scale production of food-grade DEX for use in the sugar industry.


Assuntos
Chaetomium/enzimologia , Dextranase/metabolismo , Proteínas Fúngicas/metabolismo , Cobalto/química , Dextranase/antagonistas & inibidores , Dextranase/genética , Sucos de Frutas e Vegetais/análise , Proteínas Fúngicas/antagonistas & inibidores , Proteínas Fúngicas/genética , Concentração de Íons de Hidrogênio , Cinética , Estabilidade Proteica , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Prata/química , Especificidade por Substrato , Temperatura
17.
J Zoo Wildl Med ; 51(4): 733-744, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33480553

RESUMO

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged as the cause of a global pandemic in 2019-2020. In March 2020, New York City became the epicenter in the United States for the pandemic. On 27 March 2020, a Malayan tiger (Panthera tigris jacksoni) at the Bronx Zoo in New York City developed a cough and wheezing with subsequent inappetence. Over the next week, an additional Malayan tiger and two Amur tigers (Panthera tigris altaica) in the same building and three lions (Panthera leo krugeri) in a separate building also became ill. The index case was anesthetized for diagnostic workup. Physical examination and bloodwork results were unremarkable. Thoracic radiography and ultrasonography revealed a bronchial pattern with peribronchial cuffing and mild lung consolidation with alveolar-interstitial syndrome, respectively. SARS-CoV-2 RNA was identified by real-time, reverse transcriptase PCR (rRT-PCR) on oropharyngeal and nasal swabs and tracheal wash fluid. Cytologic examination of tracheal wash fluid revealed necrosis, and viral RNA was detected in necrotic cells by in situ hybridization, confirming virus-associated tissue damage. SARS-CoV-2 was isolated from the tracheal wash fluid of the index case, as well as the feces from one Amur tiger and one lion. Fecal viral RNA shedding was confirmed in all seven clinical cases and an asymptomatic Amur tiger. Respiratory signs abated within 1-5 days for most animals, although they persisted intermittently for 16 days in the index case. Fecal RNA shedding persisted for as long as 35 days beyond cessation of respiratory signs. This case series describes the clinical presentation, diagnostic evaluation, and management of tigers and lions infected with SARS-CoV-2 and describes the duration of viral RNA fecal shedding in these cases. This report documents the first known natural transmission of SARS-CoV-2 from humans to nondomestic felids.


Assuntos
COVID-19/veterinária , Fezes/virologia , Leões/virologia , SARS-CoV-2 , Tigres/virologia , Animais , Animais de Zoológico , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/transmissão , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/isolamento & purificação , Cidade de Nova Iorque/epidemiologia , Fatores de Transcrição/genética , Fatores de Transcrição/isolamento & purificação
18.
Emerg Infect Dis ; 26(2): 374-376, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31961307

RESUMO

We report the identification of astrovirus WI65268 in a white-tailed deer with respiratory disease in the United States in 2018. This virus is a recombinant of Kagoshima1-7 and Kagoshima2-3-2 (both bovine astroviruses from Japan) and was characterized as a potential new genotype. Further surveillance of deer might help identify related isolates.


Assuntos
Infecções por Astroviridae/veterinária , Astroviridae/isolamento & purificação , Cervos , Animais , Astroviridae/genética , Infecções por Astroviridae/diagnóstico , Filogenia , Estados Unidos
19.
Emerg Infect Dis ; 26(1): 176-178, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31855534

RESUMO

We detected bovine kobuvirus (BKV) in calves with diarrhea in the United States. The strain identified is related genetically to BKVs detected in other countries. Histopathologic findings also confirmed viral infection in 2 BKV cases. Our data show BKV is a potential causative agent for diarrhea in calves.


Assuntos
Doenças dos Bovinos/virologia , Diarreia/veterinária , Kobuvirus , Infecções por Picornaviridae/veterinária , Animais , Animais Recém-Nascidos/virologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/patologia , Diarreia/epidemiologia , Diarreia/virologia , Kobuvirus/genética , Filogenia , Infecções por Picornaviridae/epidemiologia , Infecções por Picornaviridae/patologia , Infecções por Picornaviridae/virologia , Análise de Sequência de DNA , Estados Unidos/epidemiologia
20.
Emerg Infect Dis ; 26(7): 1610-1612, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32568058

RESUMO

We characterized novel coronaviruses detected in US bottlenose dolphins (BdCoVs) with diarrhea. These viruses are closely related to the other 2 known cetacean coronaviruses, Hong Kong BdCoV and beluga whale CoV. A deletion in the spike gene and insertions in the membrane gene and untranslated regions were found in US BdCoVs (unrelated to severe acute respiratory syndrome coronavirus 2).


Assuntos
Golfinho Nariz-de-Garrafa/virologia , Infecções por Coronavirus/veterinária , Diarreia/veterinária , Gammacoronavirus/classificação , Gammacoronavirus/genética , Animais , Infecções por Coronavirus/virologia , Proteínas M de Coronavírus , Diarreia/virologia , Gammacoronavirus/isolamento & purificação , Gammacoronavirus/fisiologia , Genes Virais , Genoma Viral , Mutação , Filogenia , Deleção de Sequência , Glicoproteína da Espícula de Coronavírus/genética , Proteínas da Matriz Viral/genética
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