RESUMO
Background: To date, there have been no reports assessing the incidence, risk factors, and clinical outcomes of GB disease in patients receiving ECMO for cardiorespiratory failure. Methods: The medical records of adults (aged > 18 years) who underwent ECMO between May 2010 and October 2019 were retrospectively reviewed. We investigated the prevalence and related factors of GB disease during ECMO therapy, compared clinical outcomes between patients with and without GB disease, and performed propensity-matched analysis. Results: In total, 446 patients were included, and symptomatic GB disease was found in 62 patients (13.9%, 76.2/1000 ECMO days). Complicated GB disease occurred in 42 patients (9.4%, 89.4/1000 ECMO days) and presented as acute cholecystitis, acute cholangitis, and biliary pancreatitis in 33 (7.4%), 7 (1.6%), and 5 (1.1%) patients, respectively. In multivariate Cox regression analysis, longer ECMO support (>2 weeks) (hazard ratio (HR), 2.95; 95% confidence interval (CI), 1.69−5.15) and elevated plasma hemoglobin (Hb, >50 mg/dL) (HR. 2.12; 95% CI, 1.18−3.78) were significantly associated with the development of GB disease. In the propensity-matched cohort, the intensive care unit (ICU) and hospital survival rates were significantly lower for patients with GB disease than for those without GB disease (ICU survival rate, 64.5% vs. 84.7%; hospital survival rate, 59.7% vs. 81.5%). Conclusion: The incidence of GB disease was higher in patients who received ECMO than in the general ICU patients. Furthermore, elevated plasma Hb and prolonged ECMO therapy were significant factors for the development of GB disease during ECMO therapy.
RESUMO
A/Puerto Rico/8/34 (PR8)-derived recombinant viruses have been used for seasonal flu vaccines; however, they are insufficient for vaccines against some human-fatal H5N1 highly pathogenic avian influenza (HPAI) viruses (HPAIV) due to low productivity. Additionally, the polymerase basic 2 (PB2) protein, an important mammalian-pathogenicity determinant, of PR8 possesses several mammalian-pathogenic mutations. We previously reported two avian PB2 genes (01310 and 0028) related to efficient replication in embryonated chicken eggs (ECEs) and nonpathogenicity in BALB/c mice. In this study, we generated PR8-derived H5N1 recombinant viruses harboring hemagglutinin (attenuated) and neuraminidase genes of a clade 2.3.2.1c H5N1 HPAIV (K10-483), as well as the 01310 or 0028 PB2 genes, and investigated their replication and immunogenicity. Compared with a control virus harboring six internal PR8 genes (rK10-483), the recombinant viruses possessing the 01310 and 0028 PB2 genes showed significantly higher replication efficiency in ECEs and higher antibody titers in chickens. In contrast to rK10-483, none of the viruses replicated in BALB/c mice, and all showed low titers in Madin-Darby canine kidney cells. Additionally, the recombinant viruses did not induce a neutralization antibody but elicited decreased protective immune responses against K10-483 in mice. Thus, the highly replicative and mammalian nonpathogenic recombinant H5N1 strains might be promising vaccine candidates against HPAI in poultry.
Assuntos
Virus da Influenza A Subtipo H5N1 , Vacinas contra Influenza/genética , Influenza Aviária/prevenção & controle , Infecções por Orthomyxoviridae/prevenção & controle , Animais , Galinhas/virologia , Cães , Feminino , Genes Virais/genética , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/imunologia , Vacinas contra Influenza/imunologia , Vacinas contra Influenza/uso terapêutico , Influenza Aviária/imunologia , Influenza Aviária/virologia , Células Madin Darby de Rim Canino/virologia , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , RNA Polimerase Dependente de RNA/genética , RNA Polimerase Dependente de RNA/imunologia , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/uso terapêutico , Proteínas Virais/genética , Proteínas Virais/imunologiaRESUMO
PURPOSE: Although Arg-Gly-Asp (RGD) motif-containing disintegrins are associated with integrin inhibition and the activation of various biological processes, little is known about the role of RGD motif-containing disintegrin in vascular development and remodeling. We therefore investigated the role of RGD-containing disintegrin in vascular remodeling in oxygen-induced retinopathy (OIR) mouse model. MATERIALS AND METHODS: EGT022, an RGD-containing disintegrin originated from human a disintegrin and metalloproteinase 15 (ADAM15), was used to investigate the role of the disintegrin in vascular development in OIR mouse model. To analyze the functional effects of EGT022 on retinal vascular development, the immunohistochemistry on mouse retinas after fluorescein isothiocyanate (FITC) perfusion was conducted and the vessel integrity was examined using modified Mile's permeability assay. RESULTS: EGT022 was able to reduce overall retinopathy scores by 75%, indicating its efficacy in retinal microvessel maturation stimulation. Pericyte coverage was greatly stimulated by EGT022 treatment in OIR mouse model. EGT022 was also effective to significantly improve blood vessel integrity. CONCLUSIONS: RGD-containing disintegrin EGT022 stimulated vascular maturation in OIR mouse model. Experimental results suggest that EGT022 is useful for treatments to improve ischemia in nonproliferative diabetic retinopathy (NPDR), the early stage of diabetic retinopathy.
Assuntos
Desintegrinas/farmacologia , Neovascularização Retiniana/patologia , Vasos Retinianos/patologia , Remodelação Vascular/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Oxigênio/toxicidade , Inibidores da Agregação Plaquetária/farmacologia , Vasos Retinianos/efeitos dos fármacosRESUMO
Matrix metalloproteinases (MMPs) play an important role in tissue remodeling during normal physiological situations and pathological implications such as tumor invasion and metastasis. MMP inhibitors were screened from extracts of medicinal herbs by an enzymatic assay using the MMP-14 catalytic domain. Among samples tested, a methanol extract of the root of Dalbergia odorifera T. CHEN (Leguminosae) showed the strongest inhibitory activity. The inhibitory component was purified through fractionation methods and identified as fisetin, abundant in many fruits and vegetables. In addition to inhibition of MMP-14, fisetin inhibits MMP-1, MMP-3, MMP-7, and MMP-9, more efficiently than a naturally occurring MMP inhibitor tetracycline. Fisetin dose-dependently inhibits proliferation of fibrosarcoma HT-1080 cells and human umbilical vascular endothelial cells (HUVECs), MMP-14-mediated activation of proMMP-2 in HT-1080 cells, invasiveness of HT-1080 cells, and in vitro tube formation of HUVECs. Therefore, fisetin could be valuable as a chemopreventive agent against cancer and a lead compound for development of therapeutic MMP inhibitors.
Assuntos
Flavonoides/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Inibidores de Metaloproteinases de Matriz/farmacologia , Metaloproteinases da Matriz/metabolismo , Extratos Vegetais/farmacologia , Apolipoproteínas E/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Quimioprevenção , Dalbergia/química , Relação Dose-Resposta a Droga , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Flavonóis , Gelatinases/genética , Gelatinases/metabolismo , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Invasividade Neoplásica , Raízes de Plantas/químicaRESUMO
In this study, multiplex nested RT-PCR (mnRT-PCR) was applied to simultaneous detect multiplex PCR with the higher sensitivity of nested PCR that is required for avian influenza, infectious bronchitis and Newcastle disease virus using two steps of amplification. For the first PCR, primers that were specific for each virus were newly designed from the nucleoprotein gene of AIV, the nucleocapsid protein gene of IBV and the fusion protein gene of NDV to amplify products of 665, 386 and 236 nucleotides, respectively. The multiplex PCR step provides mass amplification using common primers, which increased markedly the sensitivity of the test. Non-specific reactions were not observed when other viruses and bacteria were used for evaluating the mnRT-PCR. As a field application, 172 samples were tested by RT-PCR and mnRT-PCR. Among these samples, the concordance rates for mnRT-PCR and the single conventional RT-PCR showed 98.9% (kappa=0.98) and 98.8% (kappa=0.96) similarity for IBV and AIV, respectively. As a result, it is recommended the multiplex nested PCR as an effective tool for detecting and studying the molecular epidemiology of various mixed infections of one or more of these viruses in poultry.
Assuntos
Doenças das Aves/diagnóstico , Vírus da Bronquite Infecciosa/isolamento & purificação , Vírus da Influenza A/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Vírus da Doença de Newcastle/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Doenças das Aves/virologia , Aves , Vírus da Bronquite Infecciosa/genética , Vírus da Influenza A/genética , Vírus da Doença de Newcastle/genética , Sensibilidade e Especificidade , Medicina Veterinária/métodos , Virologia/métodos , Viroses/diagnóstico , Viroses/veterináriaRESUMO
Monoclonal antibody against kanamycin was prepared, and competitive direct ELISA and immunochromatographic assay were developed using the antibody to detect kanamycin in animal plasma and milk. The monoclonal antibody produced was identified to be IgG1, which has a kappa light chain. No cross-reactivity of the antibody was detected with other aminoglycosides, indicating that the monoclonal antibody was highly specific for kanamycin. Based on competitive direct ELISA, the detection limits of kanamycin were determined to be 1.1 ng/ml in PBS, 1.4 ng/ml in plasma, and 1.0 ng/ml in milk. The concentration of intramuscularly injected kanamycin was successfully monitored in rabbit plasma with competitive direct ELISA. Based on the colloidal gold-based immunochromatographic assay, the detection limits of kanamycin were estimated to be about 6-8 ng/ml in PBS, plasma, and milk. The immunochromatographic assay would be suitable for rapid and simple screening of kanamycin residues in veterinary medicine. Screened positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA. Therefore, the assays developed in this study could be used to complement each other as well as other laboratory findings. Moreover, instead of slaughtering the animals to obtain test samples, these methods could be applied to determine kanamycin concentration in the plasma of live animals.
Assuntos
Antibacterianos/análise , Ensaio de Imunoadsorção Enzimática/veterinária , Canamicina/análise , Leite/química , Animais , Anticorpos Monoclonais , Cromatografia/métodos , Cromatografia/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Camundongos , CoelhosRESUMO
BACKGROUND: Reliable analytical methods are required to monitor neomycin residue levels in the livestock products. In particular, a more simple and rapid detection method is required in the veterinary fields. METHODS: Competitive direct ELISA and immunochromatographic assay were developed using monoclonal antibody to detect neomycin in the animal plasma and milk. RESULTS: No cross-reactivity of the antibody was observed with other aminoglycosides based on competitive direct ELISA methods, indicating that the antibody is highly specific for neomycin. Based on the standard curves, the detection limits were determined to be 6.85 ng/ml in PBS, 3.61 ng/ml in plasma, and 2.73 ng/ml in milk, respectively. Recoveries of neomycin from spiked plasma and milk at levels of 50-200 ng/ml ranged from 87% to 108%. Concentration of intramuscularly injected neomycin was successfully monitored in the rabbit plasma through competitive direct ELISA. Immunochromatographic method was also developed using colloidal gold-conjugated monoclonal antibody. Through this method, the detection limits were estimated to be about 10 ng/ml of neomycin in PBS, plasma, and milk. CONCLUSIONS: Immunochromatographic assay developed in this study is suitable for the simple screening of neomycin residues in the veterinary field. Observed positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA.
Assuntos
Anticorpos Monoclonais/imunologia , Cromatografia/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Neomicina/análise , Animais , Especificidade de Anticorpos/imunologia , Cromatografia/instrumentação , Cromatografia/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Leite/química , Neomicina/sangue , Neomicina/imunologia , Coelhos , Reprodutibilidade dos TestesRESUMO
Competitive direct enzyme-linked immunosorbent assay (ELISA) and the immunochromatographic assay were developed using a monoclonal antibody to detect gentamicin in the animal plasma and milk. No cross-reactivity of the antibody was observed with other aminoglycosides based on competitive direct ELISA, indicating that the antibody is highly specific for gentamicin. On the basis of the standard curves, the detection limits were determined to be 0.9 ng/mL in phosphate-buffered saline (PBS), 1.0 ng/mL in plasma, and 0.5 ng/mL in milk, respectively. Recoveries of gentamicin from spiked plasma and milk at levels of 25-100 ng/mL ranged from 85 to 112%. The concentration of intramuscularly injected gentamicin was successfully monitored in the rabbit plasma through competitive direct ELISA. The detection limits were estimated to be about 6 ng/mL of gentamicin in PBS, plasma, and milk using the colloidal gold-based immunochromatographic assay, which is suitable for the simple screening of gentamicin residues in the veterinary field. Observed positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA. Therefore, the assays developed in this study could complement each other as well as veterinary field and laboratory findings.
Assuntos
Antibacterianos/análise , Cromatografia/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Gentamicinas/análise , Imunoensaio/métodos , Animais , Antibacterianos/sangue , Anticorpos Monoclonais , Feminino , Gentamicinas/sangue , Camundongos , Camundongos Endogâmicos BALB C , Leite/químicaRESUMO
We previously reported that endostatin inhibits endothelial and tumor cellular invasion by blocking activation and catalytic activity of matrix metalloproteinase (MMP)-2. Here we have examined the domain of proMMP-2 responsible for the binding of endostatin using surface plasmon resonance. ProMMP-2 and proMMP-2deltaHP lacking the hinge and hemopexin-like (HP) domains bound little to the immobilized endostatin. The active MMP-2 and MMP-2deltaHP, but not the HP domain of MMP-2, bound to endostatin at similar levels. In addition, preincubation of MMP-2 and MMP-2deltaHP with the MMP inhibitor actinonin, which binds to the active site of MMP-2, abolished their bindings to endostatin. These results indicate that endostatin binds to neither the latent proMMP-2 nor the HP domain but to the catalytic domain of MMP-2.