RESUMO
Bioremediation is a low-cost treatment alternative for the cleanup of polychlorinated-dioxin-contaminated soils and fly ash when pollution spread is wide-ranging. An interesting fungus, Ceriporia sp. MZ-340, with a high ability to degrade dioxin, was isolated from white rotten wood of a broadleaf tree from Kyushu Island in Japan. We have attempted to use the fungus for bioremediation of polychlorinated-dioxin-contaminated soil on site. However, we have to consider that this trial has the potential problem of introducing a biohazard to a natural ecosystem if this organism is naturalized. We have therefore developed a monitoring system for the introduced fungus as a part of the examination and evaluation of bioremediation in our laboratory. We have also developed a PCR-based assay to reliably detect the fungus at the bioremediation site. DNA isolated from the site was amplified by PCR using a specific primer derived from internal transcribed spacer region (ITS: ITS1, 5.8S rDNA and ITS2) sequences of Ceriporia sp. MZ-340. We successfully monitored Ceriporia sp. MZ-340 down to 100 fg/ micro l DNA and down to 2 mg/g mycelium. We also successfully monitored the fungus specifically at the bioremediation site. The polychlorinated dibenzo- p-dioxin and polychlorinated dibenzofuran content was observed to decrease in response to treatment with the fungus. The species-specific PCR technique developed in the present work is useful in evaluating the possibility of on-site bioremediation using the fungus Ceriporia sp. MZ-340.
Assuntos
Carbono , Dioxinas/metabolismo , Polyporales/isolamento & purificação , Poluentes Atmosféricos/metabolismo , Benzofuranos/análise , Biodegradação Ambiental , Carbono/análise , Carbono/química , Cinza de Carvão , DNA Fúngico/análise , DNA Espaçador Ribossômico/análise , Dioxinas/análise , Material Particulado , Reação em Cadeia da Polimerase/métodos , Polyporales/metabolismo , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: A new coating material, poly-2-methoxyethyl acrylate (PMEA), was developed to improve the biocompatibility of cardiopulmonary bypass (CPB) circuits. METHODS: To investigate the efficacy of the PMEA coating for CPB circuits, we compared PMEA-coated circuits (group P, n = 6) with uncoated circuits (group C, n = 6) and heparin (covalent-bonded heparin, Hepaface)-coated circuits (group H, n = 6) in a porcine CPB model. RESULTS: Platelet counts were significantly preserved in groups P and H compared with those in group C (P versus C, p < 0.05). The plasma levels of thrombin-antithrombin complex and bradykinin were significantly lower at 120 minutes in groups P and H than in group C (thrombin-antithrombin: P versus C, p < 0.05; bradykinin: P versus C, p < 0.05). The amount of fibrinogen adsorbed onto the hollow fibers was markedly less in group P than in groups C and H. CONCLUSIONS: The PMEA coating was equal to heparin coating in preventing reactions induced by CPB circuits, and might be superior to heparin coating in suppressing the adsorption of plasma proteins such as fibrinogen. Thus, PMEA coating may be a suitable means for improving the biocompatibility of CPB circuits.
Assuntos
Acrilatos , Ponte Cardiopulmonar/instrumentação , Materiais Revestidos Biocompatíveis , Polímeros , Animais , Antitrombina III , Bradicinina/sangue , Feminino , Fibrinogênio/metabolismo , Heparina , Teste de Materiais , Peptídeo Hidrolases/sangue , SuínosRESUMO
Mitotic cyclins A and B contain a conserved N-terminal helix upstream of the cyclin box fold that contributes to a significant interface between cyclin and cyclin-dependent kinase (CDK). To address its contribution on cyclin-CDK interaction, we have constructed mutants in conserved residues of the N-terminal helix of Xenopus cyclins B2 and A1. The mutants showed altered binding affinities to Cdc2 and/or Cdk2. We also screened for mutations in the C-terminal lobe of CDK that exhibited different binding affinities for the cyclin-CDK complex. These mutations were at residues that interact with the cyclin N-terminal helix motif. The cyclin N-terminal helix mutations have a significant effect on the interaction between the cyclin-CDK complex and specific substrates, Xenopus Cdc6 and Cdc25C. These results suggest that the N-terminal helix of mitotic cyclins is required for specific interactions with CDKs and that to interact with CDK, specific substrates Cdc6 and Cdc25C require the CDK to be associated with a cyclin. The interaction between the cyclin N-terminal helix and the CDK C-terminal lobe may contribute to binding specificity of the cyclin-CDK complex.
Assuntos
Ciclina A/metabolismo , Ciclina B/metabolismo , Quinases Ciclina-Dependentes/metabolismo , Sequência de Aminoácidos , Animais , Ciclina A/química , Ciclina B/química , Dados de Sequência Molecular , Fosforilação , Ligação Proteica , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , XenopusRESUMO
OBJECTIVE: Percutaneous cardiopulmonary support, a simplified form of venoarterial bypass, using totally heparin-coated circuits, has recently come into clinical use. To clarify its efficacy in postcardiotomy cardiogenic shock to aid weaning from cardiopulmonary bypass, we compared results of percutaneous cardiopulmonary support with those of left heart bypass using a centrifugal pump. METHODS: We reviewed 18 patients treated between 1991 and 1998 who could not be weaned from cardiopulmonary bypass. Nine were aided by totally heparin-coated percutaneous cardiopulmonary support (PCPS group), and 9 supported by left heart bypass using a centrifugal pump (LHB group). In both groups, activated clotting time was controlled at 150-200 seconds using minimal doses of heparin as needed. RESULTS: Weaning and survival rates were higher in the PCPS group than in the LHB group (100% vs 55.6%, and 66.7% vs 22.2%). The PCPS group had a smaller amount of blood loss and needed a smaller amount of blood components in the immediate postoperative period. One percutaneous cardiopulmonary support patient required surgical re-exploration for postoperative bleeding (11.1%), but no clinical thromboembolic event occurred in the PCPS group. In the LHB group, 5 patients underwent surgical re-exploration for postoperative bleeding (55.6%), and 2 underwent thrombus extirpation in the left ventricle (22.2%). CONCLUSIONS: Although this study was retrospective and historical backgrounds could have been involved, our data suggest that totally heparin-coated percutaneous cardiopulmonary support system appears more effective as an aid to weaning from cardiopulmonary bypass and in short-term circulatory support for patients in postcardiotomy cardiogenic shock.
Assuntos
Procedimentos Cirúrgicos Cardíacos , Ponte Cardiopulmonar/instrumentação , Derivação Cardíaca Esquerda , Choque Cardiogênico/terapia , Adulto , Idoso , Feminino , Heparina , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Estudos Retrospectivos , Choque Cardiogênico/cirurgia , Resultado do TratamentoRESUMO
Cardiopulmonary bypass (CPB) is known to induce an inflammatory response in association with neutrophil mediated lung injury. P-Selectin has been reported to be involved in the initiation of this inflammatory response by promoting the adhesion of neutrophils to endothelial cells in postcapillary venules. However, the role of P-selectin in the inflammatory response induced by CPB has never been clarified. To elucidate its role, we evaluated the effect of an anti-rat specific P-selectin monoclonal antibody (ARP2-4; Sumitomo Pharmaceutical) on the response of inflammatory cytokines and lung injury in a rat-CPB model. Twenty Sprague-Dawley rats underwent CPB for 30 minutes (80 ml/kg per minute, 34 degrees C) under one of two conditions. In group P, ARP2-4 (3 mg/kg) was added to the priming solution of the bypass circuit (n = 10). Saline alone was given to group C (n = 10). Inflammatory cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin[IL]-1beta, IL-6, and IL-8) and respiratory index (RI) as a marker of pulmonary gas-exchange ability were measured 1) before the initiation of CPB, 2) at the termination of CPB, and 3) 2 hours after the termination of CPB. Neither TNF-alpha nor IL-1beta was detected during the experimental period in either group. The plasma levels of IL-6 and IL-8 increased after CPB in both groups, but they were significantly lower in group P than in group C. The RI value increased in a pattern similar to that of the inflammatory cytokines and was significantly lower in group P. These data demonstrate that the addition of an anti-rat specific monoclonal antibody inhibits the abnormal release of inflammatory cytokines and attenuates CPB induced lung injury in rats. Thus, P-selectin may play a role in the augmentation of CPB induced inflammatory response, and the use of its inhibitory monoclonal antibody seems to be a promising strategy for the treatment of CPB induced lung injury.
Assuntos
Anticorpos Monoclonais/farmacologia , Ponte Cardiopulmonar/efeitos adversos , Neutrófilos/imunologia , Selectina-P/imunologia , Pneumonia/imunologia , Animais , Interleucina-1/sangue , Interleucina-1/imunologia , Interleucina-6/sangue , Interleucina-6/imunologia , Interleucina-8/sangue , Interleucina-8/imunologia , Masculino , Pneumonia/etiologia , Pneumonia/prevenção & controle , Ratos , Ratos Sprague-Dawley , Respiração , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoAssuntos
Procedimentos Cirúrgicos Cardíacos/métodos , Técnica de Fontan/efeitos adversos , Obstrução do Fluxo Ventricular Externo/cirurgia , Criança , Ventrículos do Coração/anormalidades , Humanos , Masculino , Radiografia , Obstrução do Fluxo Ventricular Externo/diagnóstico por imagem , Obstrução do Fluxo Ventricular Externo/etiologiaRESUMO
The transfer of an orally administered anti-cancer agent, UFT, into the portal vein was examined in 21 patients with hepatic metastasis of colorectal cancer (synchronous metastasis in 9 and metachronous metastasis in 12 cases) encountered at our department. The time course of tegafur, 5-FU and uracil levels in portal blood was traced for maximum 6 hrs, starting 2 hrs after the final oral dose of UFT. The portal blood tegafur level was 11.89 +/- 4.31 micrograms/ml at 2 hrs after the final dose and decreased gradually thereafter, reaching to 8.48 +/- 8.42 micrograms/ml at 6 hrs after the final dose. Unlike the portal blood tegafur level, the portal blood 5-FU level did not show any similar tendency; it remained almost unchanged at 0.018 +/- 0.006 microgram/ml and approximately equal to the serum 5-FU level throughout the observation period.