RESUMEN
Background and study aims: This study evaluated the longterm outcomes of mainly endoscopic hemostatic therapy for gastrointestinal variceal bleeding and of the transition of hemostatic therapy. Patients and methods: Among 1,163 patients treated for gastrointestinal varices between April 2006 and June 2020, a total of 125 patients who underwent emergency hemostatic therapy were enrolled. Survival rates and secondary evaluation points were analyzed. Additionally, patients were classified into two groups: the previous and latter term. Patients' background, therapeutic method, and treatment results were compared between the groups. Results: 94.4% had cirrhosis. The average Child-Pugh score was 8.90. Successful primary hemostasis rate was 98.4%, and 5.6% died within 2 weeks, all with a Child-Pugh score ≥9. The respective 1- and 5-year survival rates for Child-Pugh grade A/B were 81.3% and 55.4%, while those for Child-Pugh grade C were 58.1% and 17.8%. Child-Pugh grade C or hepatocellular carcinoma was significantly associated with poor prognosis. In total, 21.6% experienced variceal re-bleeding; 62.9% of these cases were triggered by continued alcohol consumption. There was no significant difference in survival between patients with and without variceal re-bleeding and in post-treatment survival between the previous and latter terms. In the latter term, the number of cases caused by continued alcohol consumption significantly increased. Conclusions: Multidisciplinary treatment and continuation of proper management after hemostatic therapy for variceal bleeding are crucial. Continued alcohol consumption leads to variceal bleeding and re-bleeding; its proper management, including alcohol abstinence, is one of the major challenges left in the post-directacting antivirals era.
Asunto(s)
Várices Esofágicas y Gástricas , Hemostáticos , Hepatitis C Crónica , Neoplasias Hepáticas , Várices , Antivirales , Várices Esofágicas y Gástricas/complicaciones , Várices Esofágicas y Gástricas/terapia , Hemorragia Gastrointestinal/tratamiento farmacológico , Hemorragia Gastrointestinal/etiología , Hemostasis , Hemostáticos/uso terapéutico , Hepatitis C Crónica/complicaciones , HumanosRESUMEN
BACKGROUND: There is no standard treatment available for gastric cancer patients whose sole 'non-curative factor' is positivecytological findings in peritoneal washings (CFPW). The aim of this study was to examine the safety, pharmacokinetics and efficacy for free intraperitoneal cancer cells of intraperitoneal chemotherapy with paclitaxel after gastrectomy with en bloc D2 lymph node dissection in cases of gastric cancer with positive CFPW. METHODS: Ten patients with gastric cancer who underwent gastrectomy and systemic lymphadenectomy with D2 dissection, without any other non-curative factors besides positive CFPW, were treated with early postoperative intraperitoneal paclitaxel. Intra-chemotherapeutic toxicity and operative complications were measured using NCI-CTC version 3.0. Intraperitoneal and plasma paclitaxel concentrations were measured using a high-performance liquid chromatographic assay. RESULTS: Grade 3/4 toxic effects included anemia (20%) and neutropenia (10%) that required no treatment. Operative complications were, for example, superficial surgical site infections (10%) that were treated with antibiotics. No viable cancer cells were observed in the intra-abdominal fluid 24 h after intraperitoneal administration of paclitaxel. The intraperitoneal/plasma area under the drug concentration-time curve ratio was 2,003.3:1. CONCLUSION: Intraperitoneal chemotherapy with paclitaxel is a safe and effective treatment modality for free intraperitoneal cancer cells.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antineoplásicos/administración & dosificación , Neoplasias Gástricas/tratamiento farmacológico , Adenocarcinoma/mortalidad , Adenocarcinoma/patología , Adenocarcinoma/cirugía , Anciano , Antineoplásicos/farmacocinética , Antineoplásicos/toxicidad , Protocolos de Quimioterapia Combinada Antineoplásica , Femenino , Humanos , Infusiones Parenterales , Masculino , Persona de Mediana Edad , Cavidad Peritoneal/patología , Lavado Peritoneal , Estudios Prospectivos , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/patología , Neoplasias Gástricas/cirugíaRESUMEN
BACKGROUND: FLAURA, the prospective trial of osimertinib as a first-line therapy compared with first-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), did not show superior survival benefit for osimertinib in either the subgroup of Asians or the subgroup with the L858R mutation. In addition, the superiority of osimertinib compared with second-generation EGFR-TKI is thus far unclear. PATIENTS AND METHODS: We reviewed the clinical data of all consecutive patients who were treated with osimertinib or afatinib as first-line therapy between May 2016 and October 2019 from 15 institutions in Japan. We defined the groups based on first-line EGFR-TKI as the afatinib group and the osimertinib group. Outcomes included time to discontinuation of any EGFR-TKI (TD-TKI), overall survival (OS), and time to treatment failure, with propensity score analysis carried out as an exploratory analysis in the survival and subgroup analyses. RESULTS: A total of 554 patients were enrolled. Data on 326 patients in the osimertinib group, and 224 patients in the afatinib group were analyzed. TD-TKI adjusted by propensity score in the afatinib and osimertinib groups was 18.6 months (95% confidence interval 15.8 to 22.0) and 20.5 months (95% confidence interval 13.8 to not reached), respectively, without significant difference (P = 0.204). OS adjusted by propensity score favored the afatinib group with a significant difference (P = 0.018). Subgroup analysis with propensity score showed that patients with L858R and without brain metastasis had superior survival benefit with afatinib compared with osimertinib (P < 0.001). CONCLUSIONS: TD-TKI in the afatinib group was not significantly prolonged compared with the osimertinib group in the practical data. In the exploratory analysis of patients with L858R-mutated non-small-cell lung cancer without brain metastasis, afatinib showed more benefit in OS over osimertinib.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Acrilamidas , Afatinib/uso terapéutico , Compuestos de Anilina , Protocolos de Quimioterapia Combinada Antineoplásica , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/genética , Estudios de Cohortes , Receptores ErbB/genética , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Estudios ProspectivosRESUMEN
PURPOSE: In severe acute pancreatitis (SAP), multiple organ dysfunction syndrome is a contributor to high mortality. We recently demonstrated that the serum interleukin (IL)-15 level is a predictor of the complications and mortality in clinical SAP. The aim was to investigate the role of IL-15 in experimental SAP. MATERIALS AND METHODS: SAP was induced by retrograde injection of 3 and 20% sodium deoxycholate (DCA) into biliopancreatic ducts in rats (DCA pancreatitis). Expressions of IL-15 were evaluated by Western blotting and immunohistochemical staining. Recombinant IL-15 protein was administered intraperitoneally, and the effects were investigated. RESULTS: Western blotting revealed the expressions of IL-15 in the pancreas, liver, lung and intestine in 3% DCA pancreatitis. Immunohistochemical staining showed the expression of IL-15 in the cytoplasm of each organ. In 3% DCA pancreatitis, administration of recombinant IL-15 protein attenuated the elevation of serum alanine aminotransferase (ALT) levels and improved the morphological change of the lung 18 h after the induction of SAP. Moreover, in 20% DCA pancreatitis, IL-15 improved the elevation of serum amylase and ALT levels 6 h after the induction. CONCLUSIONS: These results suggest that IL-15 is related to organ dysfunction during SAP, and that IL-15 functions as a protective factor against the organ injuries.
Asunto(s)
Interleucina-15/metabolismo , Pancreatitis/inmunología , Alanina Transaminasa/sangre , Amilasas/sangre , Animales , Ácido Desoxicólico/toxicidad , Humanos , Inmunohistoquímica , Interleucina-15/uso terapéutico , Intestino Delgado/efectos de los fármacos , Intestino Delgado/microbiología , Intestino Delgado/patología , Lesión Pulmonar/tratamiento farmacológico , Lesión Pulmonar/patología , Masculino , Pancreatitis/inducido químicamente , Pancreatitis/tratamiento farmacológico , Pancreatitis/metabolismo , Ratas , Ratas Wistar , Proteínas Recombinantes/uso terapéutico , Distribución TisularRESUMEN
This study was designed to investigate whether different vascular endothelial growth factor (VEGF) genotypes are associated with ascites formation in cirrhotic patients. Seventy cirrhotic patients were included in the study: 25 cirrhotic patients with ascites and 45 cirrhotic patients without ascites. Patient characteristics were investigated and compared between the two groups. With regard to VEGF genotype, 42 patients were C/C and 28 patients were T/T or C/T. The genotypes T/T or C/T were observed in 23 cases (51%) among the non-ascites group, but in only five cases (20%) among the ascites group. Serum levels of albumin and creatinine, and the VEGF genotypes were significantly different between the two groups. Multiple regression analysis showed that serum levels of creatinine and the VEGF genotypes were significantly correlated with ascites formation. Thus, it can be concluded that VEGF genotyping might be a valuable susceptibility marker for ascites formation in cirrhotic patients.
Asunto(s)
Ascitis/complicaciones , Ascitis/genética , Predisposición Genética a la Enfermedad , Cirrosis Hepática/complicaciones , Cirrosis Hepática/genética , Factor A de Crecimiento Endotelial Vascular/genética , Ascitis/sangre , Femenino , Humanos , Cirrosis Hepática/sangre , Masculino , Persona de Mediana Edad , Análisis de Regresión , Factor A de Crecimiento Endotelial Vascular/sangreRESUMEN
AIM: The aim of this study was to analyze restenosis after percutaneous coronary intervention, factors related to restenosis after coronary artery stenting and the degree of the risk of restenosis were evaluated. METHODS: The study enrolled 181 patients (249 lesions) who underwent the first coronary artery stenting. Multivariate analysis was performed, and the restenotic index (RI) was calculated by combining the extracted predictors. RESULTS: Among the 181 patients (249 lesions), restenosis occurred in 89 (111 lesions) and did not occur in 92 (138 lesions). Vascular revasculation was performed in 95 restenosed target lesions in 68 patients. The mean period of follow-up angiography after the procedures was 206 days in the restenosis group and 271 days in the non-restenosis group, i.e. significantly shorter in the restenosis group. As a result of multivariate analysis, diabetes mellitus, Cr level, amount of the contrast medium used and stent diameter were selected as significant factors that independently contributed to the restenosis after coronary artery stenting. By combining these factors, the RI was calculated by the following formula for the prediction of restenosis: RI=exp (1.088xCr+0.909xdiabetes mellitus+0.871xcontrast medium+0.591xstent diameter). CONCLUSION: The risk of restenosis after coronary artery stenting can be predicted to an extent according to the RI devised in this study.
Asunto(s)
Oclusión de Injerto Vascular/fisiopatología , Stents , Anciano , Angiografía Coronaria , Femenino , Humanos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Valor Predictivo de las Pruebas , Pronóstico , Medición de RiesgoRESUMEN
The effect of taxol, which is a microtubule stabilizer, was examined in a model of acute edematous pancreatitis induced in rat by the administration of caerulein. Prophylactic administration of taxol ameliorated inhibition of pancreatic secretion, increased level of serum amylase, pancreatic edema, and histological alterations in this model. Immunofluorescence studies revealed that taxol stabilized the arrangement of microtubules by the action of promoting tubulin polymerization and prevented inhibition of pancreatic digestive enzyme secretion. In isolated rat pancreatic acini, taxol reversed the inhibition of amylase secretion induced by supramaximal concentrations of cholecystokinin octapeptide and did not affect the binding of cholecystokinin octapeptide to its receptor. The results obtained in this study suggest that microtubule disorganization is the initiating event in caerulein-induced pancreatitis and that the inhibition of pancreatic digestive enzyme secretion by interfering with intracellular vesicular transport due to microtubule disorganization causes caerulein-induced pancreatitis.
Asunto(s)
Alcaloides/farmacología , Ceruletida/toxicidad , Microtúbulos/metabolismo , Pancreatitis/inducido químicamente , Enfermedad Aguda , Amilasas/sangre , Amilasas/efectos de los fármacos , Amilasas/metabolismo , Animales , Separación Celular , Edema/metabolismo , Técnica del Anticuerpo Fluorescente , Masculino , Microtúbulos/efectos de los fármacos , Paclitaxel , Páncreas/efectos de los fármacos , Páncreas/metabolismo , Páncreas/ultraestructura , Pancreatitis/prevención & control , Ratas , Receptores de Colecistoquinina/metabolismo , Sincalida/metabolismo , Sincalida/farmacología , Tripsina/análisis , Tripsina/aislamiento & purificación , Tubulina (Proteína)/análisis , Tubulina (Proteína)/aislamiento & purificaciónRESUMEN
Small amounts (0.1-0.5 mM) of deoxycholate enhanced amylase secretion, which had been induced by submaximal doses of carbachol or cholecystokinin octapeptide, without affecting the maximal levels of these reactions from isolated rat pancreatic acini. Deoxycholate alone did not induce these reactions. The other bile acids such as cholate, chenodeoxycholate, ursodeoxycholate, and taurocholate were also active. Under the similar conditions, deoxycholate enhanced the secretagogue-induced diacylglycerol formation that was derived mainly from the phospholipase C-mediated hydrolysis of phosphatidylinositol and phosphatidylinositol-4-monophosphate. Deoxycholate did not enhance the secretagogue-induced hydrolysis of phosphatidylinositol-4,5-bisphosphate or Ca2+ mobilization. Deoxycholate did not affect amylase secretion, which was induced by the simultaneous addition of protein kinase C-activating 12-O-tetradecanoylphorbol-13-acetate and Ca2+ ionophore ionomycin. Since diacylglycerol and Ca2+ may be responsible for the secretagogue-induced amylase secretion, our results indicate that small amounts of bile acids increase the sensitivity to the secretagogue of diacylglycerol formation and subsequent activation of protein kinase C, and thereby enhance amylase secretion from pancreatic acini.
Asunto(s)
Amilasas/metabolismo , Ácidos y Sales Biliares/farmacología , Páncreas/efectos de los fármacos , Fosfatidilinositoles/metabolismo , Animales , Calcio/metabolismo , Ácido Desoxicólico/farmacología , Diglicéridos/biosíntesis , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Masculino , N-Metilescopolamina , Octoxinol , Páncreas/metabolismo , Pancreatitis/etiología , Polietilenglicoles/farmacología , Ratas , Ratas Endogámicas , Derivados de Escopolamina/metabolismo , Sincalida/farmacología , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
INTRODUCTION: Intra-cardiac echocardiography (ICE) which has some benefits, can be used to obtain detailed anatomy of the heart chambers or large vessels, and the catheter positions, and it has been considered useful for improving the outcome of the ablation. In the present study, we performed pulmonary vein isolation (PVI) under real time monitoring of ICE imaging utilizing an ICE catheter placed at the junction of the left atrium (LA) and PVs (LA-PV junction). METHODS: PVI for atrial fibrillation (AF) was performed in 30 cases with drug-resistant AF (mean age: 66-years-old; including 22 males). An ICE catheter utilizing a 9 MHz frequency was inserted into the LA via the atrial septum, and placed at the LA-PV junction. Circumferential ablation was performed in the LA outside of the PV ostium, encircling both the superior and inferior ostia together under ICE imaging. RESULTS: The anatomy of the LA to the PVs and catheter sites were clearly identified by the ICE during the procedure, which enabled a precise and safe catheter manipulation with minimal fluoroscopy. Further, the wall thickness of the PV and LA, and position of the esophagus could be obtained by ICE, facilitating care in adjusting the power and/or duration of the current delivery. CONCLUSION: ICE imaging of the LA-PV junction permitted real time monitoring of the target sites for PVI during the ablation procedure, and was considered a useful technique for performing PVI.
Asunto(s)
Fibrilación Atrial/diagnóstico por imagen , Fibrilación Atrial/cirugía , Ablación por Catéter , Ecocardiografía , Venas Pulmonares/diagnóstico por imagen , Venas Pulmonares/cirugía , Anciano , Fibrilación Atrial/epidemiología , Factores de Confusión Epidemiológicos , Técnicas Electrofisiológicas Cardíacas , Femenino , Estudios de Seguimiento , Atrios Cardíacos/diagnóstico por imagen , Atrios Cardíacos/cirugía , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Persona de Mediana Edad , Tomografía Computarizada por Rayos X , Resultado del TratamientoRESUMEN
Human immunoglobulin G (IgG) anti human pancreatic elastase 1 autoantibodies were detected in sera of patients with pancreatic disorders. The characteristics of these anti elastase 1 autoantibodies and their influence on radioimmunoassay (RIA) for elastase 1 were investigated. They were placed in the IgG class by the double antibody method, and most were assumed to be of a monoclonal type from their elution profiles in gel filtration analysis. The presence of autoantibodies in serum caused an increase in apparent elastase 1 values and a decrease in the recovery of elastase 1 exogenously added to the serum. These results suggest that elastase 1 immunoassay data for autoantibody positive sera can cause misjudgement of clinical stages of patients.
Asunto(s)
Autoanticuerpos/análisis , Inmunoglobulina G/análisis , Enfermedades Pancreáticas/enzimología , Elastasa Pancreática/inmunología , Autoanticuerpos/química , Cromatografía Líquida de Alta Presión , Reacciones Falso Negativas , Humanos , Inmunoglobulina G/química , Radioisótopos de Yodo , Enfermedades Pancreáticas/patología , Elastasa Pancreática/análisis , RadioinmunoensayoRESUMEN
OBJECTIVES: We sought to assess whether oxidation products of nitric oxide (NO), nitrite (NO2-) and nitrate (NO3-), referred to as NOx, are released by the heart of patients after acute myocardial infarction (AMI) and whether NOx can be determined in peripheral blood of these patients. BACKGROUND: Previously we reported that in experimental myocardial infarction (rabbits) NOx is released mainly by inflammatory cells (macrophages) in the myocardium 3 days after onset of ischemia. NOx is formed in heart muscle from NO; NO originates through the activity of the inducible form of nitric oxide synthase (iNOS). METHODS: Eight patients with acute anterior MI and an equal number of controls were studied. Coronary venous blood was obtained by coronary sinus catheterization; NOx concentrations in coronary sinus, in arterial and peripheral venous plasma were measured. Left ventricular end-diastolic pressure was determined. Measurements were carried out 24, 48 and 72 h after onset of symptoms. The type and location of coronary arterial lesions were determined by coronary angiography. Plasma NO3- was reduced to NO2- by nitrate reductase before determination of NO2- concentration by chemiluminescence. RESULTS: The results provided evidence that in patients with acute anterior MI, the myocardial production of nitrite and nitrate (NOx) was increased, as well as the coronary arterial-venous difference. Increased NOx production by the infarcted heart accounted for the increase of NOx concentration in arterial and the peripheral venous plasma. The peak elevation of NOx occurred on days 2 and 3 after onset of the symptoms, suggesting that NOx production was at least in part the result of production of NO by inflammatory cells (macrophages) in the heart. CONCLUSIONS: The appearance of oxidative products of NO (NO2- and NO3-) in peripheral blood of patients with acute MI is the result of their increased release from infarcted heart during the inflammatory phase of myocardial ischemia. Further studies are needed to define the clinical value of these observations.
Asunto(s)
Infarto del Miocardio/metabolismo , Óxido Nítrico/metabolismo , Anciano , Anciano de 80 o más Años , Arterias , Angiografía Coronaria , Vasos Coronarios , Diástole , Femenino , Estudios de Seguimiento , Humanos , Mediciones Luminiscentes , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Infarto del Miocardio/sangre , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/patología , Miocardio/metabolismo , Miocardio/patología , Nitrato-Reductasa , Nitrato Reductasas/metabolismo , Nitratos/metabolismo , Óxido Nítrico/sangre , Óxido Nítrico Sintasa/metabolismo , Nitritos/metabolismo , Oxidación-Reducción , Venas , Función Ventricular Izquierda , Presión VentricularRESUMEN
The human heart secretes both atrial natriuretic peptide and brain natriuretic peptide. This study attempts to clarify the pathophysiological significance of the peptides in cardiovascular diseases. Using immunoradiometric assay, plasma brain natriuretic peptide and atrial natriuretic peptide levels in essential hypertension, various secondary hypertension, chronic renal failure, chronic heart failure during cardiac pacing, and acute myocardial infarction were determined. Mean plasma brain natriuretic peptide and atrial natriuretic peptide levels in healthy subjects were 3.7 +/- 0.3 and 5.7 +/- 0.3 pmol/L, respectively, and increased as a function of age. Plasma brain natriuretic peptide levels showed a larger increase than atrial natriuretic peptide levels in various cardiovascular diseases. In chronic renal failure, whereas plasma atrial natriuretic peptide levels decreased significantly after hemodialysis and were correlated with the changes in body weight, changes in plasma brain natriuretic peptide levels were less prominent and did not show such a correlation. In chronic heart failure, both basal plasma brain natriuretic peptide and atrial natriuretic peptide levels were also significantly elevated. However, in response to acute ventricular or atrial pacing, brain natriuretic peptide levels did not show any increase in contrast to the marked increase of atrial natriuretic peptide levels. In acute myocardial infarction, brain natriuretic peptide levels showed more prominent changes than atrial natriuretic peptide levels and were correlated with serum levels of creatine kinase and cardiac myosin light chain I in most patients. These results suggest that both brain and atrial natriuretic peptides play an important role in the regulation of cardiovascular homeostasis.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Envejecimiento/sangre , Factor Natriurético Atrial/sangre , Enfermedades Cardiovasculares/sangre , Proteínas del Tejido Nervioso/sangre , Neoplasias de las Glándulas Suprarrenales/sangre , Adulto , Anciano , Estimulación Cardíaca Artificial , Femenino , Insuficiencia Cardíaca/sangre , Insuficiencia Cardíaca/etiología , Humanos , Hiperaldosteronismo/sangre , Hipertensión/sangre , Fallo Renal Crónico/sangre , Fallo Renal Crónico/terapia , Masculino , Persona de Mediana Edad , Péptido Natriurético Encefálico , Feocromocitoma/sangre , Valores de Referencia , Análisis de Regresión , Diálisis RenalRESUMEN
In Swiss 3T3 cells, colon tumor-promoting deoxycholate (DOC) enhanced DNA synthesis which was induced by fibroblast growth factor (FGF) in the presence of insulin. This effect was observed only when DOC was added within 10 h after the addition of FGF. DOC by itself did not induce DNA synthesis irrespective of the presence or absence of insulin. Similar results were obtained with other colon tumor-promoting bile acids such as cholate, chenodeoxycholate and taurocholate. In contrast to these bile acids, 12-O-tetradecanoylphorbol-13-acetate (TPA) induced DNA synthesis fully without FGF in the presence of insulin. DOC did not affect TPA-induced DNA synthesis. Prolonged treatment of the cells with phorbol-12,13-dibutyrate caused the down-regulation of the phorbol ester receptor and rendered the cells unresponsive to TPA. In these cells, FGF still induced DNA synthesis in the presence of insulin, but the maximal level was reduced to about one third of that in the control cells. DOC did not enhance this DNA synthesis any more. DOC did not alter the binding of FGF to the cells. These results indicate that colon tumor-promoting bile acids enhance the mitogenic action of FGF and thereby stimulate DNA synthesis, although the phorbol ester substitutes for the mitogenic action of FGF.
Asunto(s)
Ácidos y Sales Biliares/farmacología , Carcinógenos/farmacología , Neoplasias del Colon/inducido químicamente , ADN/biosíntesis , Factores de Crecimiento de Fibroblastos/farmacología , Ésteres del Forbol/farmacología , Animales , Línea Celular , Ácido Desoxicólico/farmacología , Factores de Crecimiento de Fibroblastos/metabolismo , Insulina/farmacología , Ratones , Timidina/metabolismoRESUMEN
A factor which markedly activates Ca2+-dependent thiol protease (calpain) is associated with Triton X-100-insoluble materials, presumably structural elements such as cytoskeletons, of bovine brain microsomal fraction. This factor is extracted with 0.6 M KC1, and purified partially by sucrose density gradient centrifugation and hydroxyapatite column chromatography. The factor appears to be a heat-stable protein with an approximate Mr of 15 000. With casein as substrate this factor activates both calpain I and calpain II several-fold up to more than 10- fold without alteration of their affinity to Ca2+. Calmodulin is unable to substitute for this factor. A similar factor is associated with human platelet insoluble materials.
Asunto(s)
Química Encefálica , Calpaína/metabolismo , Microsomas/análisis , Proteínas/aislamiento & purificación , Animales , Caseínas/metabolismo , Bovinos , Centrifugación por Gradiente de Densidad , Activación Enzimática , Hidrólisis , SolubilidadRESUMEN
A small amount (50-200 microM) of deoxycholate (DOC), a colon tumor-promoting bile acid, did not show a direct effect on protein kinase C activity in a cell-free system, but enhanced fibroblast growth factor (FGF)-induced diacylglycerol formation and protein kinase C activation in Swiss 3T3 cells. DOC potentiated both reactions induced by submaximal doses of FGF but showed little effect on the maximal levels of the reactions. DOC alone was inactive in eliciting both reactions in the absence of FGF. DOC did not affect the binding of FGF to the cells. Since it has been described that diacylglycerol serves as a messenger for the activation of protein kinase C in the action of FGF in Swiss 3T3 cells [(1985) FEBS Lett. 191, 205-210], these results suggest that a small amount of DOC increases the sensitivity to FGF of diacylglycerol formation and thereby potentiates protein kinase C activation in this cell line. This action of DOC was in marked contrast to that of 12-O-tetradecanoylphorbol-13-acetate, a potent tumor-promoting phorbol ester, which directly activated protein kinase C in cell-free and intact cell systems.
Asunto(s)
Ácido Desoxicólico/farmacología , Diglicéridos/biosíntesis , Factores de Crecimiento de Fibroblastos/farmacología , Glicéridos/biosíntesis , Proteína Quinasa C/metabolismo , Animales , Línea Celular , Sistema Libre de Células , Neoplasias del Colon/inducido químicamente , Sinergismo Farmacológico , Activación Enzimática/efectos de los fármacos , Ratones , Fosforilación , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
We investigated the cytotoxicity on Madin-Darby canine kidney (MDCK) cells of pancreatitis-associated ascitic fluid (PAAF) collected from rats with experimental necrotizing pancreatitis. PAAF reduced viability of MDCK cells in a time- and dose-dependent manner. We detected DNA fragmentation on the PAAF-treated MDCK cells, indicating that the cytocidal action of PAAF is via apoptosis. From the results obtained, we conclude that PAAF contains factor(s) inducing apoptosis on MDCK cells, and we assume that apoptotic cell death is involved in the mechanism of organ failure in acute pancreatitis.
Asunto(s)
Apoptosis , Ascitis/fisiopatología , Daño del ADN , Pancreatitis/fisiopatología , Enfermedad Aguda , Animales , Línea Celular , Supervivencia Celular , ADN/química , ADN/aislamiento & purificación , Perros , Humanos , Riñón , Cinética , Masculino , Ratas , Ratas Wistar , Factores de TiempoRESUMEN
BACKGROUND: Recently renal cell apoptosis has been reported in various disorders that result in renal failure. Thus we hypothesized that renal cell injury resulting from apoptosis is involved in renal failure with severe acute pancreatitis. METHODS: Renal cell apoptosis in kidneys harvested from rats with necrotizing pancreatitis was evaluated by in situ nick-end labeling. Ascitic fluid that had been collected 6 hours after development of pancreatitis was injected into the peritoneal cavities of healthy rats, and renal apoptosis was also evaluated. The apoptosis-inducing activity of the ascitic fluid was estimated in vitro with use of isolated rat renal tubules and the normal rat kidney cell line NRK52E by nuclear staining, cell cycle analysis, and DNA electrophoresis. RESULTS: Apoptosis was detected by in situ nick-end labeling on the renal tubules 6 hours after induction of pancreatitis in vivo. Similar tubular apoptosis was detected in the rats that had intraperitoneal injection of the ascitic fluid. In in vitro analyses the ascitic fluid induced nuclear and DNA fragmentation on the isolated renal tubules and promoted apoptosis on NRK52E cells in a time-dependent manner. CONCLUSIONS: Apoptotic cell death of renal tubules occurs in severe acute pancreatitis within several hours and is possibly involved in the mechanism of renal failure through undefined substance(s) in the ascitic fluid associated with pancreatitis.
Asunto(s)
Apoptosis/fisiología , Exudados y Transudados , Túbulos Renales/patología , Pancreatitis Aguda Necrotizante/patología , Animales , Ascitis/fisiopatología , Bencimidazoles , Proteínas Sanguíneas/farmacología , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/citología , Células Epiteliales/fisiología , Colorantes Fluorescentes , Etiquetado Corte-Fin in Situ , Inyecciones Intraperitoneales , Túbulos Renales/citología , Masculino , Técnicas de Cultivo de Órganos , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/análisis , UltrafiltraciónRESUMEN
BACKGROUND: Apoptosis of hepatocytes has been reported to be involved in liver failure complicated with systemic manifestations such as endotoxemia. We hypothesized that hepatocyte apoptosis occurs in severe acute pancreatitis. METHODS: Induction of apoptosis was evaluated in the liver from rats with necrotizing pancreatitis. Apoptosis-inducing activity of the pancreatitis-associated ascitic fluid on hepatocytes was evaluated in vivo by intraperitoneal injection of the ascitic fluid and in vitro using rat primary hepatocyte culture. RESULTS: Apoptosis was detected in hepatocytes in the rats both with severe acute pancreatitis and with the intraperitoneal injection of the ascitic fluid by in situ nick-end labeling and DNA fragmentation. Apoptotic change and hepatic injury were ameliorated by administration of an interleukin-1 beta-converting enzyme inhibitor. The ascitic fluid exhibited cytocidal activity in rat primary hepatocyte culture via apoptosis, which was confirmed by DNA fragmentation, by cell cycle analysis, and by nuclear fragmentation. The neutralizing antibody for transforming growth factor-beta 1 partially blocked the apoptosis induction but the antibody to tumor necrosis factor-alpha had no effect. CONCLUSIONS: Apoptotic cell death occurs in hepatocytes in severe acute pancreatitis partially via transforming growth factor-beta 1 in the pancreatitis-associated ascitic fluid.
Asunto(s)
Apoptosis , Hígado/fisiopatología , Pancreatitis/fisiopatología , Proteínas Virales , Enfermedad Aguda , Animales , Anticuerpos/farmacología , Apoptosis/fisiología , Líquido Ascítico/fisiopatología , Ciclo Celular , Células Cultivadas , Fragmentación del ADN , Etiquetado Corte-Fin in Situ , Inyecciones Intraperitoneales , Hígado/efectos de los fármacos , Hígado/patología , Masculino , Páncreas/metabolismo , Pancreatitis/genética , Pancreatitis/patología , Ratas , Ratas Wistar , Serpinas/fisiología , Factor de Crecimiento Transformador beta/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
BACKGROUND: The mechanism of acute pancreatitis-induced hepatocellular injury is unclear. We have observed hepatocyte apoptosis in rat acute necrotizing pancreatitis. These studies were designed to determine the mediator(s) responsible for hepatocyte apoptosis and to clarify the significance of macrophages as its source. METHODS: A rat sodium deoxycholate-induced pancreatitis model was used. Immunohistochemical studies for apoptosis-inducing mediators on hepatocytes were examined in the liver and on the peritoneal macrophages. The levels of transforming growth factor-beta1 (TGF-beta1) were also evaluated quantitatively with an enzyme-linked immunosorbent assay. Induction of apoptosis on the hepatocytes was evaluated by in situ nick-end labeling and tissue DNA fragmentation enzyme-linked immunosorbent assay. Finally, the effects of TGF-beta1 neutralization and macrophage depletion were examined. RESULTS: In the liver and the peritoneal macrophages, strong expression of TGF-beta1 was detected early in the course of pancreatitis. In sodium deoxycholate-induced pancreatitis, the levels of TGF-beta1 were also elevated in the plasma (9.2 +/- 0.8 ng/mL), in the pancreatitis-associated ascitic fluid (11.5 +/- 0.6 ng/mL), and in the liver homogenate (2.8 +/- 0.3 ng/g of liver tissue). Moreover, the amount of fragmented DNA of the liver with pancreatitis was 290% +/- 20% of that with a sham operation and serum alanine aminotransferase levels elevated to 248.2 +/- 67.0 IU/L. TGF-beta1 neutralization partly blocked the positive labeling on the nuclei of the hepatocytes, the elevation of the amounts of fragmented DNA (205% +/- 10% of sham operation), and the serum alanine aminotransferase level (144.2 +/- 14.9 IU/L). On the other hand, the macrophage depletion caused a marked decrease in the TGF-beta1 protein level in the plasma (4.8 +/- 1.2 ng/mL) or in the pancreatitis-associated ascitic fluid (8.0 +/- 1.0 ng/mL). Moreover, the macrophage depletion completely inhibited the elevation of the TGF-beta1 protein level in the liver homogenate (1.5 +/- 0.4 ng/g of liver tissue), and thereafter decreased the amounts of the positive labeling on the nuclei of the hepatocytes and decreased the amount of fragmented DNA (120% +/- 18% of sham operation) and the serum alanine aminotransferase elevation (119.2 +/- 24.2 IU/L). CONCLUSIONS: In a model of sodium deoxycholate-induced pancreatitis, macrophages are responsible for pancreatitis-induced hepatocellular injury by means of apoptosis, and macrophage-derived TGF-beta1 is one of the major factors inducing the hepatocyte apoptosis.