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1.
Proc Natl Acad Sci U S A ; 121(4): e2305745121, 2024 Jan 23.
Artículo en Inglés | MEDLINE | ID: mdl-38236731

RESUMEN

The development of vaccines, which induce effective immune responses while ensuring safety and affordability, remains a substantial challenge. In this study, we proposed a vaccine model of a restructured "head-to-tail" dimer to efficiently stimulate B cell response. We also demonstrate the feasibility of using this model to develop a paramyxovirus vaccine through a low-cost rice endosperm expression system. Crystal structure and small-angle X-ray scattering data showed that the restructured hemagglutinin-neuraminidase (HN) formed tetramers with fully exposed quadruple receptor binding domains and neutralizing epitopes. In comparison with the original HN antigen and three traditional commercial whole virus vaccines, the restructured HN facilitated critical epitope exposure and initiated a faster and more potent immune response. Two-dose immunization with 0.5 µg of the restructured antigen (equivalent to one-127th of a rice grain) and one-dose with 5 µg completely protected chickens against a lethal challenge of the virus. These results demonstrate that the restructured HN from transgenic rice seeds is safe, effective, low-dose useful, and inexpensive. We provide a plant platform and a simple restructured model for highly effective vaccine development.


Asunto(s)
Oryza , Paramyxovirinae , Vacunas Virales , Animales , Pollos , Virus de la Enfermedad de Newcastle , Oryza/genética , Diseño Universal , Epítopos , Anticuerpos Antivirales
2.
Avian Pathol ; 53(2): 101-105, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38018364

RESUMEN

RESEARCH HIGHLIGHTS: A sandwich ELISA was developed to detect EDSV using the mAbs 5G4 and HRP-6G6.The sandwich ELISA maintained high specificity and sensitivity.The sandwich ELISA had equivalent consistency with real-time PCR assay.


Asunto(s)
Anticuerpos Monoclonales , Atadenovirus , Animales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Sensibilidad y Especificidad
3.
Plant Biotechnol J ; 21(12): 2546-2559, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37572354

RESUMEN

Pestiviruses, including classical swine fever virus, remain a concern for global animal health and are responsible for major economic losses of livestock worldwide. Despite high levels of vaccination, currently available commercial vaccines are limited by safety concerns, moderate efficacy, and required high doses. The development of new vaccines is therefore essential. Vaccine efforts should focus on optimizing antigen presentation to enhance immune responses. Here, we describe a simple herringbone-dimer strategy for efficient vaccine design, using the classical swine fever virus E2 expressed in a rice endosperm as an example. The expression of rE2 protein was identified, with the rE2 antigen accumulating to 480 mg/kg. Immunological assays in mice, rabbits, and pigs showed high antigenicity of rE2. Two immunizations with 284 ng of the rE2 vaccine or one shot with 5.12 µg provided effective protection in pigs without interference from pre-existing antibodies. Crystal structure and small-angle X-ray scattering results confirmed the stable herringbone dimeric conformation, which had two fully exposed duplex receptor binding domains. Our results demonstrated that rice endosperm is a promising platform for precise vaccine design, and this strategy can be universally applied to other Flaviviridae virus vaccines.


Asunto(s)
Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica , Oryza , Vacunas Virales , Animales , Porcinos , Conejos , Ratones , Peste Porcina Clásica/prevención & control , Anticuerpos Antivirales , Proteínas del Envoltorio Viral , Inmunidad
4.
BMC Geriatr ; 23(1): 728, 2023 11 09.
Artículo en Inglés | MEDLINE | ID: mdl-37946131

RESUMEN

BACKGROUND: There is a gender difference in the acceptance of osteoporosis diagnosis and treatment in patients after fragility fractures, but this difference is rarely assessed during hospitalization, and it is unclear whether these differences are age-dependent. This study aimed to evaluate the differences between male and female fragility fracture patients of different age groups who received the diagnosis and treatment of osteoporosis during hospitalization. METHODS: 31,265 fragility fracture patients aged ≥ 50 years from the Fragility Fracture Management Database in a high-volume orthopedic hospital from December 2019 to February 2023 were included in this study. We compared the differences in the rates of men and women with fragility fracture who received the measurement of bone mineral density (BMD) and bone metabolism biochemical markers (BMBMs) and treatment with anti-osteoporosis medications (AOMs), and follow-up to the internal medicine clinic within 3 months after discharge, across all age groups and across different age stages (50-59, 60-69, 70-79, and ≥ 80 years). RESULTS: The detection rates of female patients receiving BMD and BMBMs during hospitalization were 31.88% and 5.30%, respectively, compared with 22.23% and 2.69% for men. The rate of receiving any AOMs treatment was 44.63% for women and 31.60% for men. The follow-up rate of returning to the internal medicine clinic within 3 months after discharge was 9.79% for women compared to 3.00% for men. There was a significant difference between males compared to females (P < 0.0001). Analysis of patients by different age group revealed that differences in the diagnosis and treatment of osteoporosis were found only in patients under 80 years of age, while gender differences in the return to the internal medicine clinic for follow-up after discharge were present in all age groups. CONCLUSIONS: Gender differences present in osteoporosis management in patients with fragility fracture during hospitalization, especially for patients under 80 years of age. This finding suggests that orthopedic surgeons neglect to manage osteoporosis in male patients with fragility fracture during hospitalization.


Asunto(s)
Fracturas Óseas , Osteoporosis , Fracturas Osteoporóticas , Humanos , Femenino , Masculino , Anciano de 80 o más Años , Factores Sexuales , Osteoporosis/diagnóstico , Osteoporosis/epidemiología , Osteoporosis/terapia , Densidad Ósea , Hospitalización , Fracturas Osteoporóticas/diagnóstico , Fracturas Osteoporóticas/epidemiología , Fracturas Osteoporóticas/terapia
5.
Ecotoxicol Environ Saf ; 259: 115069, 2023 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-37244199

RESUMEN

Exposure to particulate matter (PM) has been associated with increased hospital admissions for influenza. Airway epithelial cells are a primary target for inhaled environmental insults including fine PM (PM2.5) and influenza viruses. The potentiation of PM2.5 exposure on the effects of influenza virus on airway epithelial cells has not been adequately elucidated. In this study, the effects of PM2.5 exposure on influenza virus (H3N2) infection and downstream modulation of inflammation and antiviral immune response were investigated using a human bronchial epithelial cell line, BEAS-2B. The results showed that PM2.5 exposure alone increased the production of pro-inflammatory cytokines including interleukin-6 (IL-6) and IL-8 but decreased the production of the antiviral cytokine interferon-ß (IFN-ß) in BEAS-2B cells while H3N2 exposure alone increased the production of IL-6, IL-8, and IFN-ß. Importantly, prior exposure to PM2.5 enhanced subsequent H3N2 infectivity, expression of viral hemagglutinin protein, as well as upregulation of IL-6 and IL-8, but reduced H3N2-induced IFN-ß production. Pre-treatment with a pharmacological inhibitor of nuclear factor-κB (NF-κB) suppressed pro-inflammatory cytokine production induced by PM2.5, H3N2, as well as PM2.5-primed H3N2 infection. Moreover, antibody-mediated neutralization of Toll-like receptor 4 (TLR4) blocked cytokine production triggered by PM2.5 or PM2.5-primed H3N2 infection, but not H3N2 alone. Taken together, exposure to PM2.5 alters H3N2-induced cytokine production and markers of replication in BEAS-2B cells, which in turn are regulated by NF-κB and TLR4.


Asunto(s)
Gripe Humana , Orthomyxoviridae , Humanos , Material Particulado/metabolismo , Receptor Toll-Like 4/metabolismo , Interleucina-6/metabolismo , FN-kappa B/metabolismo , Interleucina-8/metabolismo , Células Epiteliales , Citocinas/metabolismo , Orthomyxoviridae/metabolismo , Antivirales/metabolismo , Antivirales/farmacología
6.
Int J Mol Sci ; 24(8)2023 Apr 07.
Artículo en Inglés | MEDLINE | ID: mdl-37108056

RESUMEN

Iron is one of the essential mineral elements for the human body and this nutrient deficiency is a worldwide public health problem. Iron is essential in oxygen transport, participates in many enzyme systems in the body, and is an important trace element in maintaining basic cellular life activities. Iron also plays an important role in collagen synthesis and vitamin D metabolism. Therefore, decrease in intracellular iron can lead to disturbance in the activity and function of osteoblasts and osteoclasts, resulting in imbalance in bone homeostasis and ultimately bone loss. Indeed, iron deficiency, with or without anemia, leads to osteopenia or osteoporosis, which has been revealed by numerous clinical observations and animal studies. This review presents current knowledge on iron metabolism under iron deficiency states and the diagnosis and prevention of iron deficiency and iron deficiency anemia (IDA). With emphasis, studies related to iron deficiency and bone loss are discussed, and the potential mechanisms of iron deficiency leading to bone loss are analyzed. Finally, several measures to promote complete recovery and prevention of iron deficiency are listed to improve quality of life, including bone health.


Asunto(s)
Anemia Ferropénica , Deficiencias de Hierro , Osteoporosis , Animales , Humanos , Anemia Ferropénica/complicaciones , Anemia Ferropénica/tratamiento farmacológico , Calidad de Vida , Hierro/metabolismo , Osteoporosis/etiología , Osteoporosis/metabolismo , Factores de Riesgo
7.
Virol J ; 19(1): 121, 2022 07 22.
Artículo en Inglés | MEDLINE | ID: mdl-35869505

RESUMEN

BACKGROUND: Classical swine fever (CSF) virus is the causative agent of an economically important, highly contagious disease of pigs. CSFV is genetically and serologically related to bovine viral diarrhea virus (BVDV). BVDV infection in pigs can mimic CSF clinical signs, which cause difficulty in differentiation. Serological test for detection of virus specific antibodies is a valuable tool for diagnosis and surveillance of CSFV and BVDV infections in animals. The aim of this study was to develop the CSFV Erns and BVDV tE2 -based ELISAs to distinguishably test specific antibodies against CSFV and BVDV. METHODS: The CSFV Erns and truncated E2 (tE2, residues 690-865) of BVDV were expressed in E. coli and purified by Ni-NTA affinity chromatography, respectively. Employing Erns or tE2 protein as diagnostic antigen, indirect ELISAs were developed to distinguishably test specific antibodies against CSFV and BVDV. The specificity and sensitivity of ELISAs were evaluated using a panel of virus specific sera of pigs, immunized rabbits and immunized mice. A total 150 clinical serum samples from farm pigs were measured by the developed ELISAs and compared with virus neutralizing test (VNT). RESULTS: Indirect ELISA was established based on recombinant CSFV Erns or BVDV tE2 protein, respectively. No serological cross-reaction between antibodies against CSFV and BVDV was observed in sera of immunized rabbits, immunized mice or farm pigs by detections of the Erns and tE2 -based ELISAs. Compared to VNT, the CSFV Erns -based ELISA displayed a high sensitivity (93.3%), specificity (92.0%) and agreement rate (92.7%), and the sensitivity, specificity and agreement rate of BVDV tE2 -based ELISA was 92.3%, 95.2% and 94.7%, respectively. CONCLUSION: The newly developed ELISAs are highly specific and sensitive and would be valuable tools for serological diagnosis for CSFV and BVDV infections.


Asunto(s)
Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica , Virus de la Diarrea Viral Bovina , Vacunas Virales , Animales , Anticuerpos Antivirales , Peste Porcina Clásica/diagnóstico , Diarrea , Virus de la Diarrea Viral Bovina/genética , Ensayo de Inmunoadsorción Enzimática/métodos , Escherichia coli , Ratones , Conejos , Porcinos , Proteínas del Envoltorio Viral/genética
8.
Acta Med Okayama ; 76(6): 625-633, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36549764

RESUMEN

This investigation aimed to uncover the impact of a long noncoding RNA, SET-binding factor 2 antisense RNA1 (SBF2-AS1) on the malignant progression of gastric cancer (GC) and to further explore its underlying mechanism. SBF2-AS1 expression was quantified by qRT-PCR in GC cell lines and GC tissues. In vitro loss-of-function studies of SBF2-AS1, accompanied by flow cytometry, CCK-8, and cell invasion tests, were applied to elucidate the impact of SBF2-AS1 on the tumor progression of GC cells. Finally, Western blotting and a luciferase assay were used to detect WNT/LRP5 signaling pathway activation. SBF2-AS1 was aberrantly expressed in GC cell lines (p<0.05) and GC tissues (p<0.05). Cell invasive and proliferative capabilities were inhibited via SBF2-AS1 knockdown, resulting in apoptosis of NCI-N87 and MKN74 cells. Additionally, online database analysis uncovered a positive correlation between SBF2-AS1 and the Wnt/LRP5 signaling pathway (p<0.05). SBF2-AS1 knockdown blocked the Wnt/LRP5 signaling pathway, whereas the effects of SBF2-AS1 knockdown on the malignant genotype of MKN74 as well as NCI-N87 cells were partially restored by triggering the Wnt/ LRP5 signaling pathway. High expression of SBF2-AS1 was found in GC, the malignant progression of which was repressed via SBF2-AS1 knockdown by inhibiting the Wnt/LRP5 signaling pathway.


Asunto(s)
ARN Largo no Codificante , Neoplasias Gástricas , Vía de Señalización Wnt , Humanos , Carcinogénesis , Línea Celular Tumoral , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad/genética , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , MicroARNs/genética , ARN Largo no Codificante/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/patología , Vía de Señalización Wnt/genética
9.
New Phytol ; 229(3): 1481-1491, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-32645210

RESUMEN

Root anatomy plays important roles in the control of leaf water relations. However, few studies have evaluated whether and how anatomical traits of absorptive roots influence leaf physiology of herbaceous species in a temperate grassland. We measured absorptive root anatomical traits and leaf physiological traits of 15 herbaceous species in a temperate steppe and monitored their responses to increased precipitation in a field stimulating experiment. Root anatomical and leaf physiological traits differed among monocotyledonous grasses, monocotyledonous liliaceous species and dicotyledonous forbs. The species with higher stele: root diameter, lower root diameter and cortex thickness exhibited higher transpiration rates and stomatal conductance, but lower intrinsic water-use efficiency. Increased precipitation enhanced transpiration and stomatal conductance of forbs and lilies, but it enhanced photosynthesis in lilies exclusively. The sensitive response of lilies to precipitation may be related to their large root diameter and cortex thickness. In summary, we observed distinct differences in anatomical traits of absorptive roots among plant groups in temperate steppes. These differences drove variations in leaf physiological traits and their diverse responses to precipitation change. These findings highlight the important roles of root anatomical traits in driving leaf-level physiological processes in temperate grasslands.


Asunto(s)
Magnoliopsida , Hojas de la Planta , Fotosíntesis , Plantas , Poaceae
10.
Virol J ; 18(1): 68, 2021 04 07.
Artículo en Inglés | MEDLINE | ID: mdl-33827632

RESUMEN

BACKGROUND: H7N9 avian influenza virus (AIV) including highly and low pathogenic viruses have been detected in China since 2013. H7N9 AIV has a high mortality rate after infection in humans, and most human cases have close contacted with poultry in the live poultry market. Therefore, it is necessary to develop a rapid point-of-care testing (POCT) technique for H7N9 AIV detection. METHODS: The H7N9 AIV was inactivated and purified, and was used as the antigen to immunize BALB/c. Twelve H7-HA specific monoclonal antibodies (McAbs) were produced through the hybridoma technique. The McAb 10A8 was conjugated with colloid gold as detecting antibody; McAb 9B6 was dispensed on the nitrocellulose membran as the capture test line and the Goat-anti mouse IgG antibody was dispensed as control line respectively. The immunochromatographic strip was prepared. RESULTS: The analysis of ELISA and virus neutralization test showed that the obtained McAbs specifically recognized H7 HA. Based on the prepared strip, the detection of H7 AIV was achieved within 10 min. No cross-reaction occurred between H7 AIVs and other tested viruses. The detection limit of the strip for H7 was 2.4 log10EID50/0.1 mL for chicken swab samples. CONCLUSION: The McAbs were specific for H7 and the immunochromatographic strip developed in this study was convenient, rapid and reliable for the detection of H7 AIV. The strip could provide an effective method for the rapid and early detection of H7 AIV.


Asunto(s)
Inmunoensayo , Subtipo H7N9 del Virus de la Influenza A , Gripe Aviar , Animales , Anticuerpos Monoclonales , Cromatografía de Afinidad , Ensayo de Inmunoadsorción Enzimática , Oro Coloide , Subtipo H7N9 del Virus de la Influenza A/aislamiento & purificación , Gripe Aviar/diagnóstico , Ratones Endogámicos BALB C , Pruebas en el Punto de Atención , Aves de Corral
11.
Small ; 16(40): e2002800, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32877016

RESUMEN

Direct tracing of small extracellular vesicle (sEV) cargoes holds unprecedented importance for elucidating the mechanisms involved in intercellular communication. However, high-fidelity determination of sEVs' molecular cargoes in situ has yet to be achieved due to the difficulty in transporting molecular probes into intact sEVs. Herein, a fLuorescent Intracellular-Guided Hairpin-Tetrahedron (fLIGHT) nanoprobe is described for direct visualization of sEV microRNAs in situ. Integrating the advantages of nondestructive sEV penetration via DNA origami and single-nucleotide discrimination as well as wash-free fluorescence readout using a hairpin probe, the proposed approach enables high-fidelity fluorescence visualization of sEVs' microRNA without RNA extraction or leakage, demonstrating the potential of on-site tracing of sEV cargoes. This strategy opens an avenue to establishing universal molecular detection and labeling platforms that can facilitate both sEV-derived fundamental biological studies and molecular diagnostics.


Asunto(s)
Vesículas Extracelulares , MicroARNs , Comunicación Celular
12.
Chem Res Toxicol ; 33(9): 2351-2360, 2020 09 21.
Artículo en Inglés | MEDLINE | ID: mdl-32786540

RESUMEN

2,4-Dichlorophenol (2,4-DCP), an environmental pollutant, was reported to cause hepatotoxicity. The biochemical mechanisms of 2,4-DCP induced liver injury remain unknown. The present study showed that 2,4-DCP is chemically reactive and spontaneously reacts with GSH and bovine serum albumin to form GSH conjugates and BSA adducts. The observed conjugation/adduction apparently involved the addition of GSH and departure of chloride via the ipso substitution pathway. Two biliary GSH conjugates and one urinary N-acetyl cysteine conjugate were observed in rats given 2,4-DCP. The N-acetyl cysteine conjugate was chemically synthesized and characterized by mass spectrometry and NMR. As expected, 2,4-DCP was found to modify hepatic protein at cysteine residues in vivo by the same chemistry. The observed protein adduction reached its peak at 15 min and revealed dose dependency. The new findings allowed us to better understand the mechanisms of the toxic action of 2,4-DCP.


Asunto(s)
Clorofenoles/farmacología , Contaminantes Ambientales/farmacología , Glutatión/antagonistas & inhibidores , Albúmina Sérica Bovina/antagonistas & inhibidores , Animales , Bovinos , Clorofenoles/química , Cisteína/antagonistas & inhibidores , Cisteína/química , Contaminantes Ambientales/química , Glutatión/química , Masculino , Ratones , Ratones Endogámicos , Estructura Molecular , Ratas , Ratas Sprague-Dawley , Albúmina Sérica Bovina/química
13.
Mikrochim Acta ; 187(5): 306, 2020 04 30.
Artículo en Inglés | MEDLINE | ID: mdl-32356232

RESUMEN

A strip test is described for the optical determination of influenza virus H3 subtype. It utilizes gold nanoparticle (AuNP) coated polystyrene latex microspheres (PS) as the label and a sandwich format. The AuNP and PS particles were linked using monoclonal antibodies against influenza virus as the bridge. Under the optimal conditions, the visual detection limit of the AuNP-PS-based strip test was as low as 1/16 hemagglutination unit (HAU). It was 64 times higher than that of 10 nm (4 HAU) AuNP-based strip tests. Quantitative analysis showed that the detection limit of the AuNP-PS-based strip is 0.016 HAU. The AuNP-PS-based strip test showed no cross-reactivity to the other subtypes (H1, H5, H7, or H9) of influenza viruses. Graphical abstract .


Asunto(s)
Inmunoensayo/métodos , Virus de la Influenza A/aislamiento & purificación , Nanopartículas del Metal/química , Microesferas , Poliestirenos/química , Anticuerpos Inmovilizados/inmunología , Anticuerpos Monoclonales/inmunología , Oro/química , Inmunoensayo/instrumentación , Virus de la Influenza A/inmunología , Límite de Detección
14.
Biochem Biophys Res Commun ; 516(3): 619-625, 2019 08 27.
Artículo en Inglés | MEDLINE | ID: mdl-31235251

RESUMEN

Resistance to radiotherapy accounts for most therapeutic failures in cervical cancer patients who undergo radical radiation therapy. To indicate the possible molecular mechanism of radioresistance and improve the 5-year survival rate, we focused on how SET domain protein 3 (SETD3) regulated radioresistance in human cervical cancer cells in this study. Our results indicated that SETD3 over-expression markedly increased the radiosensitivity of cervical cancer cells with radioresistance, as evidenced by the further reduced cell viability, proliferation, DNA damage and cell death. In addition, we found that SETD3 down-regulated the expression of kinesin light chain 4 (KLC4), contributing to the radiosensitivity of cervical cancer cells, and the regulatory role of SETD3 could be abolished by KLC4 over-expression. Moreover, nitric oxide (NO) production was significantly reduced by SETD3 over-expression through repressing the expression of inducible NO synthase (iNOS) and endothelial NO synthase (eNOS) in cervical cancer cells. In vivo studies using xenograft animal models also demonstrated that SETD3 over-expression combined with irradiation treatment markedly inhibited tumor growth and induced apoptosis. In summary, our data demonstrated that down-regulated SETD3 expression markedly led to the progression of radioresistance and that promoting SETD3 expression could sensitized cervical cancer cells to radiotherapy, thereby targeting SETD3 might be a potential strategy for the clinical management of cervical cancer to improve the curative effect of radiation in cervical cancer.


Asunto(s)
Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Histona Metiltransferasas/genética , Proteínas Asociadas a Microtúbulos/genética , Tolerancia a Radiación/genética , Neoplasias del Cuello Uterino/radioterapia , Ensayos Antitumor por Modelo de Xenoinjerto/métodos , Animales , Apoptosis/genética , Apoptosis/efectos de la radiación , Línea Celular Tumoral , Supervivencia Celular/genética , Supervivencia Celular/efectos de la radiación , Daño del ADN/efectos de la radiación , Femenino , Histona Metiltransferasas/metabolismo , Humanos , Cinesinas , Ratones Endogámicos BALB C , Ratones Desnudos , Proteínas Asociadas a Microtúbulos/metabolismo , Carga Tumoral/genética , Carga Tumoral/efectos de la radiación , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/metabolismo
15.
J Neurovirol ; 25(1): 42-49, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30402823

RESUMEN

Following acute infection of mucosal surfaces by bovine herpesvirus 1 (BoHV-1), sensory neurons are a primary site for lifelong latency. Stress, as mimicked by the synthetic corticosteroid dexamethasone, consistently induces reactivation from latency. Two viral regulatory proteins (VP16 and bICP0) are expressed within 1 h after calves latently infected with BoHV-1 are treated with dexamethasone. Since the immediate early transcription unit 1 (IEtu1) promoter regulates both BoHV-1 infected cell protein 0 (bICP0) and bICP4 expressions, we hypothesized that the bICP4 protein is also expressed during early stages of reactivation from latency. In this study, we tested whether bICP4 and bICP22, the only other BoHV-1 protein known to be encoded by an immediate early gene, were expressed during reactivation from latency by generating peptide-specific antiserum to each protein. bICP4 and bICP22 protein expression were detected in trigeminal ganglionic (TG) neurons during early phases of dexamethasone-induced reactivation from latency, operationally defined as the escape from latency. Conversely, bICP4 and bICP22 were not readily detected in TG neurons of latently infected calves. In summary, it seems clear that all proteins encoded by known BoHV-1 IE genes (bICP4, bICP22, and bICP0) were expressed during early stages of dexamethasone-induced reactivation from latency.


Asunto(s)
Regulación Viral de la Expresión Génica , Herpesvirus Bovino 1/genética , Proteínas Inmediatas-Precoces/genética , Rinotraqueítis Infecciosa Bovina/virología , Células Receptoras Sensoriales/virología , Ganglio del Trigémino/virología , Proteínas Virales/genética , Animales , Anticuerpos Antivirales/química , Bovinos , Línea Celular , Dexametasona/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Células Epiteliales/virología , Herpesvirus Bovino 1/crecimiento & desarrollo , Herpesvirus Bovino 1/metabolismo , Proteínas Inmediatas-Precoces/metabolismo , Rinotraqueítis Infecciosa Bovina/patología , Riñón/efectos de los fármacos , Riñón/patología , Riñón/virología , Masculino , Células Receptoras Sensoriales/efectos de los fármacos , Células Receptoras Sensoriales/metabolismo , Células Receptoras Sensoriales/patología , Ganglio del Trigémino/efectos de los fármacos , Ganglio del Trigémino/metabolismo , Ganglio del Trigémino/patología , Proteínas Virales/metabolismo , Activación Viral/efectos de los fármacos , Latencia del Virus/efectos de los fármacos
16.
Chem Res Toxicol ; 32(9): 1791-1800, 2019 09 16.
Artículo en Inglés | MEDLINE | ID: mdl-31414593

RESUMEN

Tofacitinib (TFT) is an oral JAK inhibitor which has been approved for the treatment of moderately and severely active rheumatoid arthritis. TFT was found to show concentration-, time-, and NADPH-dependent inhibition of CYP3A4, and irreversibility of the inactivation was also observed. Incubation (40 min, 37 °C) of recombinant CYP3A4 with TFT at 200 µM resulted in >70% loss of CYP3A4 activity. Estimated kinact and KI were 0.037 min-1 and 93.2 µM, respectively. GSH and superoxide dismutase/catalase revealed minor or little protection against the CYP3A4 inactivation. Furthermore, ketoconazole attenuated TFT-mediated CYP3A4 inactivation. Epoxide and α-keto-aldehyde intermediates of TFT were trapped and characterized in microsomal incubations, respectively. The aldehyde intermediate is believed to be the key for the enzyme inactivation. Multiple P450 enzymes, including CYPs2C19, 3A4, 2D6, and 1A2, participated in the metabolism of TFT to the epoxide, while the formation of the aldehyde was mainly catalyzed by CYP3A4. In conclusion, TFT was proven to be a mechanism-based inactivator of CYP3A4.


Asunto(s)
Inhibidores del Citocromo P-450 CYP3A/farmacología , Citocromo P-450 CYP3A/metabolismo , Piperidinas/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Inhibidores del Citocromo P-450 CYP3A/síntesis química , Pruebas de Enzimas , Humanos , Cetoconazol/farmacología , Masculino , Microsomas Hepáticos/efectos de los fármacos , NADP/metabolismo , Piperidinas/síntesis química , Pirimidinas/síntesis química , Pirroles/síntesis química
17.
J Biochem Mol Toxicol ; 33(9): e22366, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31332882

RESUMEN

Colchicine (COL) is an alkaloid existing in plants of Liliaceous colchicum. It has widely been used in the treatments of many diseases, such as gout, Familial Mediterranean Fever, and tumor. However, the adverse effects of COL are an obstacle to its safe use. The present studies explored the role of metabolic demethylation in the development of COL-induced hepatotoxicity. We found that inhibition of CYP3A increased the susceptibility of mice to COL hepatotoxicity, and induction of CYP3A decreased the susceptibility of animals to the hepatotoxicity. The toxicokinetic study demonstrated that pretreatment with ketoconazole caused elevated area under the concentration-time curve of COL. Three demethylation metabolites of COL were found to be less hepatotoxic than the parent compound. It appears that the formation of electrophilic demethylation metabolites was not involved in the development of COL-induced liver injury.


Asunto(s)
Colchicina/farmacocinética , Colchicina/toxicidad , Hígado/efectos de los fármacos , Animales , Citocromo P-450 CYP3A/metabolismo , Cetoconazol/administración & dosificación , Hígado/metabolismo , Masculino , Metilación , Ratones
18.
Biologicals ; 57: 61-66, 2019 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-30477957

RESUMEN

Bovine viral diarrhea virus (BVDV) fall into cytopathic (CP) and noncytopathic (NCP) biotypes, based on their ability to kill cultured cells. NCP-BVDV can not be titrated by conventional means as used for CP-BVDV, which has impeded the identification of antiviral drugs targeting NCP-BVDV virus strains. In this study, the application of an immunoperoxidase assay in the screening of antiviral drugs was tested using two known BVDV inhibitors, ribavirin and ammonium chloride (NH4Cl). Phospholipase C inhibitor U73122 was identified to affect BVDV infection by using this immunoperoxidase assay. In addition, the results of immunoperoxidase assay were validated by real-time PCR. Taken together, the immunoperoxidase assay is a useful and versatile method suitable for antiviral drug screening targeting NCP-BVDV.


Asunto(s)
Antivirales/farmacología , Diarrea Mucosa Bovina Viral/tratamiento farmacológico , Virus de la Diarrea Viral Bovina/efectos de los fármacos , Evaluación Preclínica de Medicamentos/métodos , Técnicas para Inmunoenzimas/métodos , Cloruro de Amonio/farmacología , Animales , Diarrea Mucosa Bovina Viral/virología , Bovinos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Efecto Citopatogénico Viral/efectos de los fármacos , Virus de la Diarrea Viral Bovina/fisiología , Estrenos/farmacología , Técnicas para Inmunoenzimas/normas , Pirrolidinonas/farmacología , Ribavirina/farmacología , Replicación Viral/efectos de los fármacos
19.
Drug Dev Res ; 80(8): 1062-1070, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31396981

RESUMEN

To study the efficacy of a polyoxometalate, Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O, as an antiviral treatment in HBV transgenic mice. HBV transgenic mice were treated with Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O by intragastric administration. Adefovir and distilled water were administered as controls. Serum HBV DNA, liver HBV RNA levels were measured by quantitative RT-PCR. Serum HBsAg levels were measured by ELISA. The hepatitis B virus surface antigen (HBsAg) in liver cells was detected by immunohistochemistry (IHC). Pathological changes in the liver tissues were also observed by light and electron microscopy. Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O significantly decreased serum HBsAg and HBV DNA levels. Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O resulted in a 98% decrease in serum HBV DNA at 28 days, from 4.3 log10 copies/ml at baseline to 2.5 log10 copies/ml after treatment, and the inhibition rate of HBV DNA was higher than ADV at the same dose. The HBV replication levels in each group slightly increased at 7 days after withdrawal, but rebounded slightly more in the Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O treatment group compared to the H2 O control group (p < .05). There were no differences in HBV RNA levels. No significant differences were observed in the pathology, but there were decreased HBsAg levels in the Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O-treated group compared to the control group. The results demonstrated that Cs2 K4 Na[SiW9 Nb3 O40 ]·H2 O displayed potent anti-HBV activity in HBV transgenic mice and supported for future clinic study.


Asunto(s)
Antivirales/administración & dosificación , Virus de la Hepatitis B/efectos de los fármacos , Hepatitis B/tratamiento farmacológico , Compuestos de Tungsteno/administración & dosificación , Animales , Antivirales/farmacología , ADN Viral/efectos de los fármacos , ADN Viral/genética , Modelos Animales de Enfermedad , Femenino , Hepatitis B/genética , Hepatitis B/virología , Antígenos de Superficie de la Hepatitis B/metabolismo , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/fisiología , Hígado/metabolismo , Masculino , Ratones , Ratones Transgénicos , Distribución Aleatoria , Compuestos de Tungsteno/farmacología , Replicación Viral/efectos de los fármacos
20.
BMC Genomics ; 19(1): 866, 2018 Dec 04.
Artículo en Inglés | MEDLINE | ID: mdl-30509164

RESUMEN

BACKGROUND: The goat is an important farm animal. Reproduction is an important process of goat farming. The ovary is the most important reproductive organ for goats. In recent years, an increasing number of long non-coding RNAs (lncRNAs) have been implicated in the regulation of mammal reproduction. However, there are few studies on the function of lncRNAs in reproduction, particularly lncRNAs in the ovary. RESULTS: The sequencing of goat ovaries generated 1,122,014,112 clean reads, and 4926 lncRNAs and 1454 TUCPs (transcripts of uncertain coding potential) were identified for further analysis by using the coding potential analysis software, CNCI, CPC and Pfam-sca. There were 115 /22 differential lncRNAs /TUCPs transcripts between the ovaries of the luteal phase and the follicular phase. We predicted the related genes of lncRNA /TUCP based on co-expression and co-localization methods. In total, 2584 /904 genes were predicted by co-expression, and 326/73 genes were predicted by co-localization. The functions of these genes were further analyzed with GO and KEGG analysis. The results showed that lncRNAs /TUCPs, which are highly expressed in goat ovaries in the luteal phase, are mainly associated with the synthesis of progesterone, and we filtered the lncRNAs /TUCPs, such as XR_001918177.1 and TUCP_001362, which may regulate the synthesis of progesterone; lncRNAs /TUCPs, which are highly expressed in goat ovaries in the follicular phase, are mainly associated with oogenesis and the maturation of oocytes, and we filtered the lncRNAs /TUCPs that may regulate the oogenesis and maturation of oocyte, such as XR_001917388.1 and TUCP_000849. CONCLUSION: The present study provided the genome expression profile of lncRNAs /TUCPs in goat ovaries at different estrus periods and filtered the potential lncRNAs /TUCPs associated with goat reproduction. These results are helpful to further study the molecular mechanisms of goat reproduction.


Asunto(s)
Cabras/genética , Ovario/metabolismo , ARN Largo no Codificante/metabolismo , Animales , Femenino , Fase Folicular/genética , Estudio de Asociación del Genoma Completo , Fase Luteínica/genética , Progesterona/biosíntesis , ARN/química , ARN/aislamiento & purificación , ARN/metabolismo , Análisis de Secuencia de ARN , Transcriptoma
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