Association between blood caspase-8 levels and mortality of patients with malignant middle cerebral artery infarction.

OBJECTIVE: High concentrations of caspase-8 (main initiator caspase of apoptosis extrinsic pathway) have been found in brain tissue from traumatic brain injury patients and in blood of patients with different diseases. However, there are not data on blood caspase-8 concentrations in ischemic stroke patients. Therefore, the objective of this study was to determine whether there is an association between blood caspase-8 concentrations and the probability and speed of mortality at 30 days in patients with malignant middle cerebral artery infarction (MMCAI). DESIGN: Observational prospective study. SETTING: Five Intensive Care Units (ICU). PATIENTS: Patients with severe malignant middle cerebral artery infarction (MMCAI) defined as acute infarction in more than of 50% of that territory and Glasgow Coma Scale (GCS)<9. INTERVENTIONS: Determination of serum caspase-8 levels when MMCAI was diagnosed. MAIN VARIABLES OF INTEREST: Mortality at 30 days and time until this event. RESULTS: Severe MMCAI patients (n=28) compared to survivor patients (n=28) showed higher serum caspase-8 concentrations (p<0.001), lower platelet count (p=0.01) and lower GCS (p=0.002). We found an area under the curve for mortality prediction of 78% (95% CI=65%-91%; p<0.001) by serum caspase-8 levels. Kaplan-Meier analysis found higher mortality rate in patients with serum caspase-8 levels >62.8ng/mL (hazard ratio=11.2; 95% CI=4.4-28.4; p<0.001). CONCLUSIONS: The association of high blood caspase-8 concentrations with the rate and the velocity of 30-day mortality in MMCAI patients is the main new finding of our study.

Association between blood caspase-8 levels and mortality of patients with malignant middle cerebral artery infarction / Asociación de niveles sanguíneos de caspasa-8 y mortalidad de pacientes con infarto maligno de la arteria cerebral media

Objective High concentrations of caspase-8 (main initiator caspase of apoptosis extrinsic pathway) have been found in brain tissue from traumatic brain injury patients and in blood of patients with different diseases. However, there are not data on blood caspase-8 concentrations in ischemic stroke patients. Therefore, the objective of this study was to determine whether there is an association between blood caspase-8 concentrations and the probability and speed of mortality at 30 days in patients with malignant middle cerebral artery infarction (MMCAI). Design Observational prospective study. Setting Five Intensive Care Units (ICU). Patients Patients with severe malignant middle cerebral artery infarction (MMCAI) defined as acute infarction in more than of 50% of that territory and Glasgow Coma Scale (GCS)<9. Interventions Determination of serum caspase-8 levels when MMCAI was diagnosed. Main variables of interest Mortality at 30 days and time until this event. Results Severe MMCAI patients (n=28) compared to survivor patients (n=28) showed higher serum caspase-8 concentrations (p<0.001), lower platelet count (p=0.01) and lower GCS (p=0.002). We found an area under the curve for mortality prediction of 78% (95% CI=65%–91%; p<0.001) by serum caspase-8 levels. Kaplan–Meier analysis found higher mortality rate in patients with serum caspase-8 levels >62.8ng/mL (hazard ratio=11.2; 95% CI=4.4–28.4; p<0.001). Conclusions The association of high blood caspase-8 concentrations with the rate and the velocity of 30-day mortality in MMCAI patients is the main new finding of our study (AU)

Objetivo Se han encontrado altas concentraciones de caspasa-8 (principal caspasa iniciadora de la vía extrínseca de apoptosis) en el tejido cerebral de pacientes con traumatismo craneoencefálico y en la sangre de pacientes con diferentes enfermedades. Sin embargo, no hay datos sobre las concentraciones sanguíneas de caspasa-8 en pacientes con ictus isquémico. Por tanto, el objetivo de este estudio fue determinar si existe una asociación entre las concentraciones sanguíneas de caspasa-8 y la probabilidad y velocidad de mortalidad a 30días en pacientes con infarto maligno de la arteria cerebral media (MMCAI). Diseño Observacional y prospectivo. Ámbito Cinco unidades de cuidados intensivos (UCI). Pacientes Pacientes con MMCAI grave definido como infarto agudo en más del 50% de ese territorio y escala de coma de Glasgow (GCS)<9. Intervenciones Determinación de niveles séricos de caspasa-8 cuando se diagnosticó el MMCAI grave. Variables de interés principal Mortalidad hasta los 30dias y tiempo hasta este evento. Resultados Los pacientes fallecidos (n=28) en comparación con los supervivientes (n=28) mostraron mayores concentraciones séricas de caspasa-8 (p<0,001), menor recuento plaquetario (p=0,01) y menor GCS (p=0,002). Encontramos un área bajo la curva para la predicción de mortalidad del 78% (IC 95%: 65-91%; p<0,001) por los niveles séricos de caspasa-8. El análisis de Kaplan-Meier encontró una mayor tasa de mortalidad en pacientes con niveles séricos de caspasa-8>62,8ng/mL (hazard ratio: 11,2; IC 95%: 4,4-28,4; p<0,001). Conclusiones La asociación de elevadas concentraciones sanguíneas de caspasa-8 con la tasa y velocidad de mortalidad a 30días en pacientes con MMCAI es el principal hallazgo nuevo de nuestro estudio (AU)

Blood caspase-8 concentrations and mortality among septic patients / Niveles sanguíneos de caspasa-8 y mortalidad de los pacientes sépticos

Objective: No data are available on blood caspase-8 concentrations (the initiator caspase in the extrinsic apoptosis pathway) in septic patients. The present study thus describes the blood caspase-8 concentrations in survivors and non-survivors, and examines the possible association between blood caspase-8 concentrations and mortality in septic patients.Design: A prospective observational study was carried out.Setting: Three Spanish Intensive Care Units.Patients: Septic patients.Interventions: Serum caspase-8 concentrations were determined at the diagnosis of sepsis.Main variable of interest: Mortality after 30 days.Results: Patients not surviving at day 30 (n=81) compared to surviving patients (n=140) showed higher serum caspase-8 levels (p<0.001). Multiple logistic regression analysis found an association between serum caspase-8 levels>43.5ng/ml and mortality (OR=3.306; 95%CI=1.619-6.753; p=0.001). The area under the curve (AUC) for mortality predicted by serum caspase-8 levels was 67% (95% CI=60-73%; p<0.001).Conclusions: The novel findings of our study were that blood caspase-8 concentrations are higher in non-survivors than in survivors, and that there is an association between blood caspase-8 concentrations and mortality in septic patients (AU)

Objetivo: No existen datos publicados sobre los niveles sanguíneos de caspasa-8 (la caspasa iniciadora en la vía extrínseca de apoptosis) en pacientes sépticos. Por lo tanto, los objetivos del estudio fueron describir los niveles sanguíneos de caspasa-8 en pacientes supervivientes y fallecidos y determinar si existe una asociación entre los niveles sanguíneos de caspasa-8 y la mortalidad de los pacientes sépticos.DiseñoEstudio observacional y prospectivo.ÁmbitoTres unidades de cuidados intensivos españolas.PacientesPacientes sépticos.IntervencionesSe determinaron las concentraciones séricas de caspasa-8 al diagnóstico de la sepsis.Variable de interés principalMortalidad a los 30 días.ResultadosEncontramos que los pacientes fallecidos en los primeros 30 días (n=81) comparados con los pacientes supervivientes (n=140) presentaban niveles séricos mayores de caspasa-8 (p<0,001). En el análisis de regresión logística múltiple encontramos una asociación entre los niveles séricos de caspasa-8>43,5ng/ml y la mortalidad (OR: 3,306; IC 95%: 1,619-6,753; p=0,001). El área bajo la curva para predecir la mortalidad por los niveles séricos de caspasa-8 fue del 67% (IC 95%: 60-73%; p<0,001).ConclusionesLos nuevos hallazgos de nuestro estudio fueron que los niveles séricos mayores de caspasa-8 eran superiores en los pacientes fallecidos en los primeros 30 días, y que existe una asociación entre los niveles séricos de caspasa-8 y la mortalidad (AU)

Blood caspase-8 concentrations and mortality among septic patients.

OBJECTIVE: No data are available on blood caspase-8 concentrations (the initiator caspase in the extrinsic apoptosis pathway) in septic patients. The present study thus describes the blood caspase-8 concentrations in survivors and non-survivors, and examines the possible association between blood caspase-8 concentrations and mortality in septic patients. DESIGN: A prospective observational study was carried out. SETTING: Three Spanish Intensive Care Units. PATIENTS: Septic patients. INTERVENTIONS: Serum caspase-8 concentrations were determined at the diagnosis of sepsis. MAIN VARIABLE OF INTEREST: Mortality after 30 days. RESULTS: Patients not surviving at day 30 (n=81) compared to surviving patients (n=140) showed higher serum caspase-8 levels (p<0.001). Multiple logistic regression analysis found an association between serum caspase-8 levels>43.5ng/ml and mortality (OR=3.306; 95%CI=1.619-6.753; p=0.001). The area under the curve (AUC) for mortality predicted by serum caspase-8 levels was 67% (95% CI=60-73%; p<0.001). CONCLUSIONS: The novel findings of our study were that blood caspase-8 concentrations are higher in non-survivors than in survivors, and that there is an association between blood caspase-8 concentrations and mortality in septic patients.

Association between blood caspase-8 levels and mortality of patients with malignant middle cerebral artery infarction.

OBJECTIVE: High concentrations of caspase-8 (main initiator caspase of apoptosis extrinsic pathway) have been found in brain tissue from traumatic brain injury patients and in blood of patients with different diseases. However, there are not data on blood caspase-8 concentrations in ischemic stroke patients. Therefore, the objective of this study was to determine whether there is an association between blood caspase-8 concentrations and the probability and speed of mortality at 30 days in patients with malignant middle cerebral artery infarction (MMCAI). DESIGN: Observational prospective study. SETTING: Five Intensive Care Units (ICU). PATIENTS: Patients with severe malignant middle cerebral artery infarction (MMCAI) defined as acute infarction in more than of 50% of that territory and Glasgow Coma Scale (GCS)<9. INTERVENTIONS: Determination of serum caspase-8 levels when MMCAI was diagnosed. MAIN VARIABLES OF INTEREST: Mortality at 30 days and time until this event. RESULTS: Severe MMCAI patients (n=28) compared to survivor patients (n=28) showed higher serum caspase-8 concentrations (p<0.001), lower platelet count (p=0.01) and lower GCS (p=0.002). We found an area under the curve for mortality prediction of 78% (95% CI=65%-91%; p<0.001) by serum caspase-8 levels. Kaplan-Meier analysis found higher mortality rate in patients with serum caspase-8 levels >62.8ng/mL (hazard ratio=11.2; 95% CI=4.4-28.4; p<0.001). CONCLUSIONS: The association of high blood caspase-8 concentrations with the rate and the velocity of 30-day mortality in MMCAI patients is the main new finding of our study.

Blood caspase-8 concentrations and mortality among septic patients.

OBJECTIVE: No data are available on blood caspase-8 concentrations (the initiator caspase in the extrinsic apoptosis pathway) in septic patients. The present study thus describes the blood caspase-8 concentrations in survivors and non-survivors, and examines the possible association between blood caspase-8 concentrations and mortality in septic patients. DESIGN: A prospective observational study was carried out. SETTING: Three Spanish Intensive Care Units. PATIENTS: Septic patients. INTERVENTIONS: Serum caspase-8 concentrations were determined at the diagnosis of sepsis. MAIN VARIABLE OF INTEREST: Mortality after 30 days. RESULTS: Patients not surviving at day 30 (n=81) compared to surviving patients (n=140) showed higher serum caspase-8 levels (p<0.001). Multiple logistic regression analysis found an association between serum caspase-8 levels>43.5ng/ml and mortality (OR=3.306; 95%CI=1.619-6.753; p=0.001). The area under the curve (AUC) for mortality predicted by serum caspase-8 levels was 67% (95% CI=60-73%; p<0.001). CONCLUSIONS: The novel findings of our study were that blood caspase-8 concentrations are higher in non-survivors than in survivors, and that there is an association between blood caspase-8 concentrations and mortality in septic patients.

Immunohistochemistry analysis of proteins related with apoptosis as prognostic factor in epidermoid carcinomaof penis / Análisis inmunohistoquímico de proteínas relacionadas con la apoptosis como factor pronóstico en el carcinoma epidermoide de pene

The aim was to analyze the protein expression of apoptotic genes caspase-3, caspase-8 and bcl-2 with the immunohistochemistry technique, correlating with tumor grade (I, II and III) and with the patient survival in order to understand the basic mechanism of tumoral transformation. The immunohistochemistry reactions on 50 samples of squamous cell carcinoma were carried out with the avidin-biotin immunoperoxidase method and antigen recovery. The analyses were made using the graduation method "in crosses" (0 to 4 crosses - no stain to more than 75 percent of positives cells) and in categories (low, intermediate, high) of the cytoplasm immunoreactivity of the epidermoid penile carcinoma cells. It was observed a statistically significant difference when the expression of caspase-3 were compared with the grades I and II of the tumor (p=0.0010) and when comparing the patient survival with the grades I and II of the tumor (p=0.0212). The protein bcl-2 was more expressed than caspase-3 and caspase-8 proteins, suggesting that the apoptotic rate in this carcinoma is low. The higher expression of the anti-apoptotic protein bcl-2 suggests a higher preservation of the tumoral cells...

El objetivo fue analizar la expresión de las proteínas de genes de apoptosis caspasa-3, caspasa-8 y Bcl-2-con la técnica de inmunohistoquímica, en correlación con el grado tumoral (I, II y III) y la supervivencia del paciente con el fin de comprender el mecanismo básico de la transformación tumoral. Se analizaron las reacciones inmunohistoquímicas sobre 50 muestras de carcinoma de células escamosas mediante el método de la inmunoperoxidasa avidina-biotina y la recuperación de antígeno. Los análisis se realizaron utilizando el método de graduación "en cruces" (0 a 4 cruces - no tinción a más del 75 por ciento de las células positivas) y en categorías (baja, media, alta) de la inmunorreactividad citoplasmática de las células de carcinoma epidermoide de pene. Se observó una diferencia estadísticamente significativa cuando la expresión de la caspasa-3 se comparó con los grados I y II del tumor (p = 0,0010) y cuando se comparan la supervivencia de los pacientes con los grados I y II del tumor (p = 0,0212). La proteína bcl-2 se expresa más que la caspasa-3 y caspasa-8, lo que sugiere que la tasa de apoptosis en este carcinoma es baja. La mayor expresión de la proteína anti-apoptótica bcl-2 sugiere una mayor preservación de las células tumorales...

Cassia alata leaf extract induces cytotoxicity in A549 lung cancer cells via a mechanism that is caspase 8 dependent / El extracto de la hoja de cassia alata induce citotoxicidad en las células A549 del cáncer pulmonar humano a través de un mecanismo dependiente de la caspasa 8

OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.

OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.