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1.
J Invertebr Pathol ; 206: 108162, 2024 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-38944151

RESUMO

Decapod iridescent virus 1 (DIV1) stands as a significant pathogen affecting crustaceans, posing a grave threat to the shrimp industries in aquaculture dependent nations. Within the Iridoviridae family, the conserved envelope protein DIV1-168L plays a pivotal role in virion entry. Nonetheless, the host factors that interact with 168L remain unidentified. To address this gap, we established a cDNA library derived from Litopenaeus vannamei gill tissue and conducted yeast two-hybrid screening to identify host factors that interact with 168L. Additionally, we performed co-immunoprecipitation assays to verify the interaction between cuticle protein 8 (CP8) and 168L. Expression pattern analysis revealed the presence of CP8 transcripts in the gill and epidermis. Furthermore, immunohistochemistry results demonstrated the expression of CP8 in gill cells and its localization in the gill filament epithelium. Fluorescence analysis indicated that full-length CP8 colocalized with 168L in the cytoplasm of Sf9 cells. Removal of the signal peptide from the N-terminal of CP8 eliminated its concentration in the cytoplasm. Additionally, CP8 expression was significantly inhibited during DIV1 infection. Therefore, our research contributes to a better understanding of the entry mechanism of iridovirids. The GenBank accession number for the DIV1 sequence is MF197913.1.

2.
Toxicol Appl Pharmacol ; 461: 116401, 2023 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-36706924

RESUMO

Chlorophenols (CPs) are widespread pollutants in nature. CPs have raised significant concern due to their potential hepatotoxic effects on humans. This research aimed to ascertain the inhibitory potential of eleven CPs (2-CP, 3-CP, 4-CP, 2,4-DCP, 2,3,4-TCP, 2,4,5-TCP, 2,4,6-TCP, 2,3,4,5-TeCP, 2,3,4,6-TeCP, 2,3,5,6-TeCP, and PCP) on nine human CYP isoforms (CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4). The CPs that inhibit the activity of CYP isoforms were detected with human liver microsomes (HLM) using a cocktail approach in vitro. The results demonstrated that trichlorophenols, tetrachlorophenols, and PCP strongly inhibited CYP2C8 and CYP2C9. The half inhibition concentration (IC50) value of 2,3,4,6-TeCP and PCP for CYP2C8 inhibition was 27.3 µM and 12.3 µM, respectively. The IC50 for the inhibition of 2,4,6-TCP, 2,3,4,6-TeCP and PCP towards CYP2C9 were calculated to be 30.3 µM, 5.8 µM and 2.2 µM, respectively. 2,3,4,6-TeCP, and PCP exhibited non-competitive inhibition towards CYP2C8. 2,4,6-TCP, 2,3,4,6-TeCP, and PCP exhibited competitive inhibition towards CYP2C9. The inhibition kinetics parameters (Ki) were 51.51 µM, 22.28 µM, 37.86 µM, 7.27 µM, 0.68 µM for 2,3,4,6-TeCP-CYP2C8, PCP-CYP2C8, 2,4,6-TCP-CYP2C9, 2,3,4,6-TeCP-CYP2C9, PCP-CYP2C9, respectively. This study also defined clear structure-activity relationships (SAR) of CPs on CYP2C8, supported by molecular docking studies. Overall, CPs were found to cause inhibitory effects on CYP isoforms in vitro, and this finding may provide a basis for CPs focused on CYP isoforms inhibition endpoints.


Assuntos
Clorofenóis , Inibidores das Enzimas do Citocromo P-450 , Humanos , Citocromo P-450 CYP2C8 , Citocromo P-450 CYP2C9/farmacologia , Simulação de Acoplamento Molecular , Inibidores das Enzimas do Citocromo P-450/toxicidade , Sistema Enzimático do Citocromo P-450 , Microssomos Hepáticos , Clorofenóis/toxicidade
3.
Br J Clin Pharmacol ; 89(6): 1862-1872, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36662574

RESUMO

AIMS: Apatinib is widely used in Chinese cancer patients. As the in vivo drug disposition of apatinib has large individual differences, adverse events are prone to occur. Cytochrome P450 (CYP)3A5 and cancer types maybe the main factors affecting this individual differences. The objective of our study was to establish a population pharmacokinetics (PK) model of apatinib in adult cancer patients, and to explore optimal dosage regimens for individualized treatment. METHODS: Adult patients with various types of cancer treated with apatinib were enrolled. The concentration of apatinib in plasma was determined by high-performance liquid chromatography-tandem mass spectrometry. CYP3A5 genotype was determined using TaqMan allelic discrimination technique. The population PK model was developed by NONMEM V7.4. The dosing regimen was optimized based on Monte Carlo simulations. RESULTS: A population PK model of apatinib in adult cancer patient was established. CYP3A5 genotype and systemic cancer type (digestive system cancers, nondigestive system cancers) were the most significant covariates for PK parameters. Patients with CYP3A5*1 expressers (CYP3A5*1/*1 and CYP3A5*1/*3) had lower apparent clearance and apparent volume of distribution than patients who do not express CYP3A5*1 (CYP3A5*3/*3). Patients with nondigestive system cancer had higher apparent volume of distribution and absorption rate constant than digestive system cancer. The results of dose simulation suggest that the apatinib dose in patients who do not express CYP3A5*1 should be 33.33-50.00% higher than that in CYP3A5*1 expressers. CONCLUSIONS: A population PK model of apatinib in adult cancer patients was established. CYP3A5 genotype and systemic cancer type had concurrent effects on PK parameters. CYP3A5 patients who do not express CYP3A5*1 required higher doses.


Assuntos
Citocromo P-450 CYP3A , Neoplasias , Humanos , Adulto , Citocromo P-450 CYP3A/genética , Farmacogenética , Neoplasias/tratamento farmacológico , Neoplasias/genética , Piridinas/efeitos adversos , Genótipo , Imunossupressores , Tacrolimo
4.
Clin Chem Lab Med ; 61(6): 1069-1074, 2023 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-36635945

RESUMO

OBJECTIVES: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become a common technique in clinical laboratories in recent years. Because most methods are laboratory-developed tests (LDTs), their reproducibility and quality control (QC) have been controversial. In this study, Westgard Sigma Rules were used to evaluate the analytical performance and establish an individualised internal QC (IQC) strategy for these LDTs. METHODS: Taking the LC-MS/MS LDT method for homocysteine (Hcy) as an example, the 'desirable specifications' from the Biological Variation Database were used as quality goals. Based on the external quality assessment (EQA) samples, bias was calculated and the coefficient of variation (CV) was also calculated by IQC measurements for six consecutive months. The analytical performance was evaluated by calculated sigma metrics and an IQC strategy was designed using the Westgard Sigma Rules with run size. RESULTS: Over 116 days within 6 months, a total of 850 data points were collected for each of IQC 1 and IQC 2. The monthly coefficient of variation CV% was 2.57-4.01%, which was non-significant (p-value: 0.75). The absolute bias% for IQC1 and IQC2 was 1.23 and 1.87%, respectively. The allowable total error (TEa) was selected as 15.5%, Sigma metrics were 4.02 and 4.30, and the analytical performance was 'Good'. The 13s/22s/R4s/41s multi rules (n=4, r=1) with a run size of 200 samples were suggested for the Hcy IQC scheme. The quality goal index (QGI) values were over 1.2, indicating that trueness needed to be improved. CONCLUSIONS: The analytical performance of the Hcy LC-MS/MS LDT conformed to the Six Sigma rating level, achieving 'good' (four Sigma). Clinical practice indicated that calibration bias was the primary factor affecting trueness.


Assuntos
Espectrometria de Massas em Tandem , Gestão da Qualidade Total , Humanos , Cromatografia Líquida , Reprodutibilidade dos Testes , Controle de Qualidade , Gestão da Qualidade Total/métodos
5.
Anal Bioanal Chem ; 415(12): 2281-2289, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36952025

RESUMO

Cadmium ion (Cd(II)) is a pernicious environmental pollutant that has been shown to contaminate agricultural lands, accumulate through the food chain, and seriously threaten human health. At present, Cd(II) monitoring is dependent on centralized instruments, necessitating the development of rapid and on-site detection platforms. Against this backdrop, the present study reports on the development of a fluorometric aptasensor designed to target Cd(II), which is achieved through the integration of strand displacement amplification (SDA) and CRISPR/Cas12a. In the absence of Cd(II), the aptamer initiates SDA, resulting in the generation of a profusion of ssDNA that activates Cas12a, leading to a substantial increase in fluorescence output. Conversely, the presence of Cd(II) curtails the SDA efficiency, culminating in a significant reduction in fluorescence output. The proposed approach has been demonstrated to enable the selective detection of Cd(II) at concentrations of 60 pM, with the performance of the aptasensor validated in real water and rice samples. The proposed platform based on aptamer-target interaction holds immense promise as a signal-amplified and precise method for the detection of Cd(II) and has the potential to transform current hazard detection practices in food samples.


Assuntos
Técnicas Biossensoriais , Poluentes Ambientais , Humanos , Sistemas CRISPR-Cas , Cádmio , Agricultura , DNA de Cadeia Simples , Oligonucleotídeos
6.
Biomed Chromatogr ; 37(6): e5625, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36919355

RESUMO

Liver cirrhosis is currently the twelfth leading cause of death globally and the sixth leading cause of death in China. Its treatment is expensive. Changes in the composition of the serum bile acid pool are sensitive indicators of the severity of liver cirrhosis. In this study, a novel LC-tandem mass spectrometry (LC-MS/MS) method was developed and used to simultaneously determine 15 bile acids in human serum in patients with decompensated cirrhosis. Sample preparation involved spiking with isotope internal standards followed by protein precipitation. The analytical run time was 5 min. The LC-MS/MS method was fully validated according to Clinical and Laboratory Standards Institute (CLSI) C62A and the consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories. The method achieved an acceptable coefficient of variation for precision (0.83%-14.80%) and accuracy (89.39%-107.62%). Finally, as proof of applicability, the method was applied to patients with decompensated cirrhosis in routine clinical sample analysis. The degree of variation of different bile acids was clearly shown. These results indicated that abnormal metabolic pathways might play important roles in decompensated cirrhosis.


Assuntos
Ácidos e Sais Biliares , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida/métodos , Padrões de Referência , Reprodutibilidade dos Testes
7.
J Environ Manage ; 325(Pt A): 116372, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36252327

RESUMO

Although lakes dominated by macrophytes are conducive to ecological balance, this balance is easily disrupted by excessive nutrients flowing into the lake. However, knowledge of whether excessive nutrients lead to different microbial environmental vulnerabilities in the lake sediment between macrophyte-dominated areas and macrophyte-free areas is a prerequisite for the implementation of targeted protection measures. In this study, we investigated bacterial communities in sediments using high-throughput sequencing of 16S rRNA genes. Our results showed that the sources of total nitrogen (TN) and organic matter (OM) were related to the macrophytes. The structure, drivers, and interspecific associations of bacterial community, which were more susceptible to increased changes in TN and OM, differed significantly between macrophyte-dominated areas and macrophyte-free areas. More precisely, the lake edge, where was occupied by macrophytes, had a higher proportion of deterministic phylogenetic turnover (88.89%) than other sites, as well as a wider ecological niche and a tighter network structure. Further, as the difference in TN increased, the main assembly processes in surface sediments changed from stochastic to deterministic. However, the majority of phyla from the lake edge showed a greater correlation with excessive nutrients, and the selection of the community by excessive nutrients was more obvious at the edge of the lake. In addition, our results demonstrated that the stability of the bacterial community in macrophyte-free areas is greater than in macrophyte-dominated areas, while an excessively high deterministic process ratio and nutrient (TN and OM) concentration significantly reduced bacterial community stability at macrophyte-dominated areas. Taken together, these results provide a better understanding of the effects of excessive nutrients derived from macrophytes on bacterial community patterns, and highlight the importance of avoiding the accumulation of TN and OM in macrophyte-dominated areas to enhance the sustainability of the ecosystem after restoration of lakes with macrophytes.


Assuntos
Lagos , Microbiota , Ecossistema , Sedimentos Geológicos/microbiologia , RNA Ribossômico 16S/genética , Filogenia , Nitrogênio , Bactérias/genética , Nutrientes , China , Fósforo
8.
J Clin Biochem Nutr ; 73(2): 161-171, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37700848

RESUMO

Metabolic differences between colorectal cancer (CRC) and NI (NI) play an important role in early diagnoses and in-time treatments. We investigated the metabolic alterations between CRC patients and NI, and identified some potential biomarkers, and these biomarkers might be used as indicators for diagnosis of CRC. In this study, there were 79 NI, 50 CRC I patients, 52 CRC II patients, 56 CRC III patients, and 52 CRC IV patients. MS-MS was used to measure the metabolic alterations. Univariate and multivariate data analysis and metabolic pathway analysis were applied to analyze metabolic data and determine differential metabolites. These indicators revealed that amino acid and fatty acids could separate these groups. Several metabolites indicated an excellent variables capability in the separation of CRC patients and NI. Ornithine, arginine, octadecanoyl carnitine, palmitoyl carnitine, adipoyl carnitine, and butyryl carnitine/propanoyl carnitine were selected to distinguish the CRC patients and NI. And methionine and propanoyl carnitine, were directly linked to different stages of CRC. Receiver operating characteristics curves and variables importance in projection both represented an excellent performance of these metabolites. In conclusion, we assessed the difference between CRC patients and NI, which supports guidelines for an early diagnosis and effective treatment.

9.
J Immunol ; 205(8): 2207-2221, 2020 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-32917788

RESUMO

Foot-and-mouth disease is a highly contagious disease of pigs, sheep, goats, bovine, and various wild cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV) that has given rise to significant economic loss to global livestock industry. FMDV 3B protein is an important determinant of virulence of the virus. Modifications in 3B protein of FMDV considerably decrease virus yield. In the current study, we demonstrated the significant role of 3B protein in suppression of type I IFN production and host antiviral response in both human embryonic kidney HEK293T cells and porcine kidney PK-15 cells. We found that 3B protein interacted with the viral RNA sensor RIG-I to block RIG-I-mediated immune signaling. 3B protein did not affect the expression of RIG-I but interacted with RIG-I to block the interaction between RIG-I and the E3 ubiquitin ligase TRIM25, which prevented the TRIM25-mediated, Lys63-linked ubiquitination and activation of RIG-I. This inhibition of RIG-I-mediated immune signaling by 3B protein decreased IFN-ß, IFN-stimulated genes, and proinflammatory cytokines expression, which in turn promoted FMDV replication. All of the three nonidentical copies of 3B could inhibit type I IFN production, and the aa 17A in each copy of 3B was involved in suppression of IFN-related antiviral response during FMDV infection in porcine cells. Together, our results indicate the role of 3B in suppression of host innate immune response and reveal a novel antagonistic mechanism of FMDV that is mediated by 3B protein.


Assuntos
Proteína DEAD-box 58/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/imunologia , Imunidade Inata , Receptores Imunológicos/imunologia , Transdução de Sinais/imunologia , Animais , Células HEK293 , Humanos , Interferon beta/imunologia , Suínos , Fatores de Transcrição/imunologia , Proteínas com Motivo Tripartido/imunologia , Ubiquitina-Proteína Ligases/imunologia , Ubiquitinação/imunologia
10.
Biomed Chromatogr ; 36(6): e5366, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35274340

RESUMO

Schizophrenia is a debilitating psychiatric disorder affecting approximately 1% of the population worldwide. Disturbances in the homocysteine metabolism are an important factor in the pathophysiology of schizophrenia. In this research, a novel validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantification procedure was developed to investigate three significant compounds of homocysteine metabolism: homocysteine, cysteine, and methionine in patients with schizophrenia and healthy controls. Sample preparation involved a reduction with dithiothreitol followed by protein precipitation, and the chromatographic runtime was 2 min. The LC-MS/MS method was validated according to CLSI C62-A and the Chinese Guidance for Liquid Chromatography and Mass Spectrometry Clinical Application. The performance of the method was excellent with a coefficient of variation for precision in the range of 0.5-6.9%, an accuracy of 90.4-101.6%. In addition, the practical applicability of the method was demonstrated by applying it in the routine sample analysis for a schizophrenic patient. Increased homocysteine levels and decreased cysteine levels were observed in the patient with schizophrenia. These results indicate that the activity of the transsulfuration pathway may play a key role in the pathogenesis of schizophrenia.


Assuntos
Esquizofrenia , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Cisteína/química , Homocisteína , Humanos , Metionina , Espectrometria de Massas em Tandem/métodos
11.
Environ Sci Technol ; 54(23): 15097-15107, 2020 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-33167623

RESUMO

This study reports on the compositional diversity of organic compounds in metal(loid)-bearing tailings samples from both active and abandoned tailings ponds. Tailings samples were qualitatively analyzed by comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOFMS). In addition, the priority PAHs (16), PAEs (6), and phenols (2) were quantitatively analyzed using gas chromatography-mass spectrometry (GC-MS). We attribute the presence of some of aromatic organics in studied tailings ponds to particular sources. Mineral floatation reagents are likely the major sources of small-ring aromatics in tailings ponds, and products from metallurgical processing and burning of fossil fuels in the mining area or further afield are also possible contributors and might be the main source of large-ring aromatics. We found that tailings ponds abandoned for decades can still have organics concentrations at levels of concern. Large-ring aromatics are generally more toxic than other contaminants, and these were more abundant in abandoned tailings ponds. This suggests that these large-ring organics do not readily decompose or biodegrade into less toxic byproducts, as do volatiles and many other organic compounds. Our aromatic contaminants database provides an important starting point for researchers to investigate and compare similar contaminants that might be also present in other tailings ponds and emphasizes the necessity of considering their transformations over time.


Assuntos
Hidrocarbonetos Policíclicos Aromáticos , Lagoas , Metais/análise , Mineração , Compostos Orgânicos , Hidrocarbonetos Policíclicos Aromáticos/análise
12.
J Med Virol ; 91(2): 208-214, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30039874

RESUMO

Foot-and-mouth disease (FMD) is a disease of worldwide economic importance, and vaccines play an important role in preventing FMDV outbreaks. However, new control strategies are still needed since FMDV outbreaks still occur in some disease-free countries. Currently, interferon (IFN)-based strategies have been demonstrated to be an efficient biotherapeutic option against FMDV; however, interferon omega (IFN-ω) has not yet been assessed in this capacity. Thus, this study evaluated the antiviral activity of porcine IFN omega 7 (PoIFN-ω7) against FMDV. After the PoIFN-ω7 was expressed and purified, cell proliferation assays and quantitative real-time reverse transciption-polymerase chain reaction were used to evaluate the effective anti-cytopathic concentration of PoIFN-ω7 and its effectiveness pre- and post-infection with FMDV in swine kidney cells (IBRS-2). Results showed the rHis-PoIFN-ω7 fusion protein was considerably expressed using Escherichia coli BL21 (DE3) strain, and the recombinant protein exhibited significant in vitro protection against FMDV, including two strains belonging to type O and A FMDV, respectively. In addition, PoIFN-ω7 upregulated the transcription of Mx1, ISG15, OAS1, and PKR genes. These findings indicated that IFN-ω has the potential for serving as a useful therapeutic agent to prevent FMDV or other viral outbreaks in pigs.


Assuntos
Antivirais/farmacologia , Vírus da Febre Aftosa/efeitos dos fármacos , Vírus da Febre Aftosa/crescimento & desenvolvimento , Interferon Tipo I/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Animais , Efeito Citopatogênico Viral , Interferon Tipo I/genética , Proteínas Recombinantes de Fusão/genética , Suínos
13.
J Med Virol ; 91(12): 2142-2152, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31347713

RESUMO

Recently, many countries, including China, have experienced a series of type A and O foot-and-mouth disease virus (FMDV) epidemics, causing serious economic losses. Although concerns about the safety of inactivated FMD vaccines have been raised, the development of a safe and effective subunit vaccine is necessary. We constructed two chimeric virus-like particles (VLPs; rHBc/AO and rHBc/AOT VLPs) displaying tandem repeats of B cell epitopes (VP1 residue 134-161 and 200-213) derived from type A and O FMDV and one T cell epitope (3 A residue 21-35) using the truncated hepatitis B virus core (HBc) carrier. Our results indicate that the chimeric HBc can self-assemble into VLPs with these FMDV epitopes displayed on the surface. Immunization with the chimeric VLPs induced specific IgG and neutralization antibodies against type A and O FMDV in mice. Compared with the commercial type A/O FMDV bivalent inactivated vaccine, rHBc/AO and rHBc/AOT VLPs significantly stimulated the production of Th1 type cytokines (IFN-γ and IL-2), whereas Th2 cytokine production (IL-4 and IL-10) was decreased. Compared with rHBc/AO, rHBc/AOT induced increased Th2 cytokine and specific IgG production. These results demonstrate that the VLPs constructed in the current study induced both humoral and cellular immune responses and may represent potential bivalent VLP vaccines targeting both FMDV type A and O strains.


Assuntos
Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Vírus da Hepatite B/imunologia , Proteínas do Core Viral/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Citocinas/imunologia , Feminino , Vírus da Febre Aftosa/química , Vírus da Hepatite B/química , Imunoglobulina G/sangue , Camundongos , Organismos Livres de Patógenos Específicos , Células Th1/imunologia , Células Th2/imunologia , Vacinação , Vacinas de Partículas Semelhantes a Vírus/genética , Vacinas de Partículas Semelhantes a Vírus/imunologia , Proteínas do Core Viral/química
14.
J Med Virol ; 91(9): 1595-1601, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31032977

RESUMO

Foot-and-mouth disease (FMD) is a highly contagious disease that affects cloven-hoof animals including cattle, swine, sheep, goats, and lots of wild species. Effectively control measures are urged needed. Here, we showed that homoharringtonine treatment exhibited a strong inhibitory effect against two different strains of FMDVs (O/MYA98/BY/2010 and A/GD/MM/2013) in swine kidney (IBRS-2) cells. Further experiments demonstrated that homoharringtonine did not affect virus attachment or entry. Using time-of-addition assays, we found that the antiviral activity of homoharringtonine occurred primarily during the early stage of infection. These results demonstrated that homoharringtonine might be an effective anti-FMDV drug. Further studies are required to explore the antiviral activity of homoharringtonine against FMDV replication in vivo.


Assuntos
Antivirais/farmacologia , Vírus da Febre Aftosa/efeitos dos fármacos , Febre Aftosa/virologia , Mepesuccinato de Omacetaxina/farmacologia , Animais , Antivirais/química , Linhagem Celular , Vírus da Febre Aftosa/fisiologia , Mepesuccinato de Omacetaxina/química , Humanos , Estrutura Molecular , Internalização do Vírus , Replicação Viral/efeitos dos fármacos
15.
Microb Pathog ; 127: 79-84, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30500407

RESUMO

Recently, a novel type I interferon alphaomega (IFN-αω), also known as IFN-µ, was identified. However, the biological activity of IFN-αω remain poorly understood. In this study, the porcine IFN-αω (PoIFN-αω) was expressed, purified, and its antiviral activities assessed by its ability to inhibit the cytopathic effect caused by FMDV on IBRS-2 cells. In addition, q-PCR was used to evaluate the expression of IFN-stimulated genes induced by PoIFN-αω. It was found that PoIFN-αω exerted effective antiviral activity against FMDV pre- and post-infection. Additionally, PoIFN-αω induced the transcription of IFN-stimulated genes, including Mx1, ISG15, OAS1, and PKR genes. Our study reported a new indication of PoIFN-αω as an effective anti-FMDV agent for the first time.


Assuntos
Antivirais/farmacologia , Vírus da Febre Aftosa/efeitos dos fármacos , Interferon Tipo I/farmacologia , Proteínas Recombinantes/farmacologia , Animais , Antivirais/isolamento & purificação , Antivirais/metabolismo , Linhagem Celular , Efeito Citopatogênico Viral , Perfilação da Expressão Gênica , Fatores Imunológicos/biossíntese , Interferon Tipo I/genética , Interferon Tipo I/isolamento & purificação , Interferon Tipo I/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Suínos
16.
Cell Physiol Biochem ; 51(5): 2377-2396, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30537741

RESUMO

The interferons (IFNs) are a primary defense against pathogens because of the strong antiviral activities they induce. IFNs can be classified into three groups: type I, type II and type III, according to their genetic, structural, and functional characteristics and their receptors on the cell surface. The type I IFNs are the largest group and include IFN-α, IFN-ß, IFN-ε, IFN-ω, IFN-κ, IFN-δ, IFN-τ and IFN-ζ. The use of IFNs for the treatment of viral infectious diseases on their antiviral activity may become an important therapeutic option, for example, IFN-α is well known for the successful treatment of hepatitis B and C virus infections, and interest is increasing in the antiviral efficacy of other novel IFN classes and their potential applications. Therefore, in this review, we summarize the recent progress in the study of the biological activities of all the type I IFN classes and their potential applications in the treatment of infections with immunodeficiency virus, hepatitis viruses, and influenza viruses.


Assuntos
Antivirais/uso terapêutico , Infecções por HIV/tratamento farmacológico , Hepatite Viral Humana/tratamento farmacológico , Influenza Humana/tratamento farmacológico , Interferon Tipo I/uso terapêutico , Animais , Antivirais/farmacologia , HIV/efeitos dos fármacos , Vírus de Hepatite/efeitos dos fármacos , Humanos , Interferon Tipo I/farmacologia , Orthomyxoviridae/efeitos dos fármacos , Infecções por Orthomyxoviridae/tratamento farmacológico
17.
J Med Virol ; 89(11): 2041-2046, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28390158

RESUMO

Foot-and-mouth disease virus (FMDV) causes an economically important and highly contagious disease of cloven-hoofed animals such as cattle, swine, and sheep. FMD vaccine is the traditional way to protect against the disease, which can greatly reduce its occurrence. However, the use of FMD vaccines to protect early infection is limited. Therefore, the alternative strategy of applying antiviral agents is required to control the spread of FMDV in outbreak situations. As previously reported, LiCl has obviously inhibition effects on a variety of viruses such as transmissible gastroenteritis virus (TGEV), infectious bronchitis coronavirus (IBV), and pseudorabies herpesvirus and EV-A71 virus. In this study, our findings were the first to demonstrate that LiCl inhibition of the FMDV replication. In this study, BHK-21 cell was dose-dependent with LiCl at various stages of FMDV. Virus titration assay was calculated by the 50% tissue culture infected dose (TCID50 ) with the Reed and Muench method. The cytotoxicity assay of LiCl was performed by the CCK8 kit. The expression level of viral mRNA was measured by RT-qPCR. The results revealed LiCl can inhibit FMDV replication, but it cannot affect FMDV attachment stage and entry stage in the course of FMDV life cycle. Further studies confirmed that the LiCl affect the replication stage of FMDV, especially the early stages of FMDV replication. So LiCl has potential as an effective anti-FMDV drug. Therefore, LiCl may be an effective drug for the control of FMDV. Based on that, the mechanism of the antiviral effect of LiCl on FMDV infection is need to in-depth research in vivo.


Assuntos
Antivirais/farmacologia , Vírus da Febre Aftosa/efeitos dos fármacos , Cloreto de Lítio/farmacologia , Replicação Viral/efeitos dos fármacos , Animais , Bovinos , Linhagem Celular , Replicação do DNA/efeitos dos fármacos , Febre Aftosa/tratamento farmacológico , Febre Aftosa/virologia , Vírus da Febre Aftosa/genética , Vírus da Febre Aftosa/fisiologia , Reação em Cadeia da Polimerase , RNA Viral/genética , Ovinos , Suínos , Fatores de Tempo
18.
BMC Genomics ; 17(1): 865, 2016 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-27809759

RESUMO

BACKGROUND: Gram-negative bacteria of the genus Serratia are potential producers of many useful secondary metabolites, such as prodigiosin and serrawettins, which have potential applications in environmental bioremediation or in the pharmaceutical industry. Several Serratia strains produce prodigiosin and serrawettin W1 as the main bioactive compounds, and the biosynthetic pathways are co-regulated by quorum sensing (QS). In contrast, the Serratia strain, which can simultaneously produce prodigiosin and serrawettin W2, has not been reported. This study focused on analyzing the genomic sequence of Serratia sp. strain YD25T isolated from rhizosphere soil under continuously planted burley tobacco collected from Yongding, Fujian province, China, which is unique in producing both prodigiosin and serrawettin W2. RESULTS: A hybrid polyketide synthases (PKS)-non-ribosomal peptide synthetases (NRPS) gene cluster putatively involved in biosynthesis of antimicrobial serrawettin W2 was identified in the genome of YD25T, and its biosynthesis pathway was proposed. We found potent antimicrobial activity of serrawettin W2 purified from YD25T against various pathogenic bacteria and fungi as well as antitumor activity against Hela cells. Subsequently, comparative genomic analyses were performed among a total of 133 Serratia species. The prodigiosin biosynthesis gene cluster in YD25T belongs to the type I pig cluster, which is the main form of pig-encoding genes existing in most of the pigmented Serratia species. In addition, a complete autoinducer-2 (AI-2) system (including luxS, lsrBACDEF, lsrGK, and lsrR) as a conserved bacterial operator is found in the genome of Serratia sp. strain YD25T. Phylogenetic analysis based on concatenated Lsr and LuxS proteins revealed that YD25T formed an independent branch and was clearly distant from the strains that solely produce either prodigiosin or serrawettin W2. The Fe (III) ion reduction assay confirmed that strain YD25T could produce an AI-2 signal molecule. Phylogenetic analysis using the genomic sequence of YD25T combined with phylogenetic and phenotypic analyses support this strain as a member of a novel and previously uncharacterized Serratia species. CONCLUSION: Genomic sequence and metabolite analysis of Serratia surfactantfaciens YD25T indicate that this strain can be further explored for the production of useful metabolites. Unveiling the genomic sequence of S. surfactantfaciens YD25T benefits the usage of this unique strain as a model system for studying the biosynthesis regulation of both prodigiosin and serrawettin W2 by the QS system.


Assuntos
Genoma Bacteriano , Genômica , Lipoproteínas/biossíntese , Metaboloma , Metabolômica , Peptídeos Cíclicos/biossíntese , Prodigiosina/biossíntese , Serratia/genética , Serratia/metabolismo , Antibacterianos/biossíntese , Antibacterianos/farmacologia , Biologia Computacional/métodos , Mineração de Dados , Ácidos Graxos/metabolismo , Genômica/métodos , Lipoproteínas/genética , Lipoproteínas/farmacologia , Metabolômica/métodos , Família Multigênica , Peptídeos Cíclicos/genética , Peptídeos Cíclicos/farmacologia , Fenótipo , Filogenia , Prodigiosina/farmacologia , Percepção de Quorum/genética , Serratia/classificação
19.
J Med Virol ; 87(8): 1436-40, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25952001

RESUMO

Since 1997, more and more cases of the infectious H5N1 avian influenza virus (AIV) in humans have been reported all over the world but the transmission of H5N1 avian influenza virus to stray cats has been little demonstrated. The objective of this pilot investigation was to determine the prevalence of H5N1 AIV antibodies in stray cats in eastern China where is the dominant enzootic H5N1 highly pathogenic avian influenza virus (HP AIV). A total of 1,020 nasal swab and 1,020 serum samples were collected and tested. Evidence of HPAI H5N1 virus antibodies was present in two of the 1,020 serum samples that were positive by HI assay and NT assay, respectively. The results imply little transmission and that the Clade 2.3.2 HPAIV H5N1 infections in poultry did not significantly affect the rural animal shelters or suburban environment in eastern China. In future studies, these results can be used as baseline seroepidemiological levels for H5N1 AIV among cats in China.


Assuntos
Anticorpos Antivirais/sangue , Doenças do Gato/epidemiologia , Doenças do Gato/virologia , Virus da Influenza A Subtipo H5N1/imunologia , Infecções por Orthomyxoviridae/veterinária , Animais , Gatos , China/epidemiologia , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Projetos Piloto , Estudos Soroepidemiológicos
20.
Microb Pathog ; 80: 63-6, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25680835

RESUMO

In this study, we sought to examine whether evidence existed suggesting that pigs were being infected with the novel H7N9 avian influenza virus. From November 2012 to November 2013, blood was drawn from 1560 pigs from 100 large farms in 4 provinces of eastern China. Many of these pigs were in close proximity to wild birds or poultry. Swine sera were studied using hemagglutinin inhibition (HI) assays and enzyme-linked immunosorbent assays (ELISAs) against the H7 antigen derived from the emergent H7N9 avian influenza virus (AIV). Only 29 of the 1560 samples had HI titers of 1:20 when using the H7N9 AIV antigens, and none of the 29 (H7N9 AIV) HI-positive samples were positive when using ELISA, indicating that no samples were positive for H7N9. The negative results were also verified using a novel competitive HA-ELISA. As pigs have been shown to be infected with other avian influenza viruses and as the prevalence of novel influenza A viruses (e.g., H7N9 AIV) may be increasing among poultry in China, similar seroepidemiological studies of pigs should be periodically conducted in the future.


Assuntos
Anticorpos Antivirais/sangue , Subtipo H7N9 do Vírus da Influenza A/imunologia , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/virologia , Animais , Aves , China/epidemiologia , Ensaio de Imunoadsorção Enzimática , Testes de Inibição da Hemaglutinação , Influenza Aviária/transmissão , Infecções por Orthomyxoviridae/diagnóstico , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/epidemiologia
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