RESUMEN
The objectives of this study are to investigate the apoptotic changes in gastric adenocarcinoma following neoadjuvant chemotherapy and to illuminate its correlation with response. One 67 gastric cancer patients with cT2-4 or TanyN1-3M0 between January 2006 and December 2007 were included. All patients had previous gastrectomy and D2 lymphadenectomy with curative intent performed. A total of 12 cycles of preoperative mFOLFOX7 chemotherapy was recommended for all patients, and 83 patients received only adjuvant chemotherapy. Resected specimens were subjected to in situ TUNEL assay and scanned with Applied Imaging Ariol SL-50. Apoptosis index (AI) was significantly higher in the patient received preoperative chemotherapy (CS group) than in the patient did not (S group). In the CS group, AI was found to have a strong positive correlation with the pathological response (rho = 0.403, P = 0.0002). A ROC curve presented a score of 49.4 as the AI cutoff value for response, dividing the CS group into two subgroups with significantly different prognoses (P = 0.003) and further allowing identification of 8 patients with significantly better prognosis out of 48 patients evaluated as grades 1a-b according to a pathological response evaluation (P = 0.022). In conclusion, AI is correlated with efficacy of preoperative chemotherapy and prognosis of gastric cancer patients following neoadjuvant chemotherapy. An AI cutoff value for response may be used as a complementary approach to current pathological response evaluations to help identify potential responders.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Apoptosis/efectos de los fármacos , Neoplasias Gástricas/tratamiento farmacológico , Adenocarcinoma/patología , Adenocarcinoma/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Fluorouracilo/administración & dosificación , Humanos , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Estimación de Kaplan-Meier , Leucovorina/administración & dosificación , Masculino , Persona de Mediana Edad , Análisis Multivariante , Estadificación de Neoplasias , Compuestos Organoplatinos/administración & dosificación , Oxaliplatino , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Neoplasias Gástricas/patología , Neoplasias Gástricas/cirugía , Resultado del TratamientoRESUMEN
PURPOSE: Fecal incontinence and constipation still remain as major postoperative complications after procedures for anorectal malformations (ARM). The striated muscle complex (SMC) is one of the most important factors that influence defecation. Previous studies have demonstrated different degrees of the muscle complex dysplasia dependent on the complexity of ARM. To explore the mechanisms of maldevelopment of SMC in ARM, apoptosis was investigated during pelvic floor muscle development in rat embryos with ARM. METHODS: Anorectal malformations in rat embryos were induced by treating pregnant rats with ethylenethiourea on the 10th embryonic day (E10). Normal and ARM rat embryos from E16 to E21 were serial-sectioned transversely or sagittally, and SMCs were dissected and snap frozen. TdT mediated dUTP Nick Ending Labeling (TUNEL) staining and DNA ladder analysis were performed to identify apoptosis and expression of Bax/Bcl-2 were confirmed with immunohistochemical staining and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) analysis. RESULTS: Hypoplastic and disordered SMC sling shifted cephalad, ventrally, and converged inferior to the rectourethral fistula and infiltrated connective tissue in ARM embryos. In the normal group, TUNEL-positive cells became evident on E17; sporadic positive staining was mainly localized in 2 areas as follows: the junction area between SMC and bulbocarvernosus muscle and posterior to the rectum where bilateral SMC converged. In the ARM group, massive positive staining of nuclei was observed from E16 to E21 and was mainly distributed in the dorsal part of the SMC. Electrophoresis of DNA samples yielded a "ladder" pattern of migration both in normal and the ARM group from E17 to E21, the ladders were stronger in the ARM group. In both groups, the expression of Bax/Bcl-2 was detectable on E17, the immunoreactivity increased on E19 and E21. Compared with the normal group, the expression of Bax was increased, whereas Bcl-2 was declined in the ARM group. Significant upregulation of Bax messenger RNA (mRNA) levels and downregulation of Bcl-2 mRNA levels were observed in ARM embryos. CONCLUSIONS: In the current study, abnormal apoptosis and disturbed expression of Bax/Bcl-2 were identified during SMC development in ARM embryos. It is suggested that precocious, excessive, and dislocated apoptosis might be a fundamental pathogenesis for the maldeveloped SMC in ARM rats. The temporospatial expressions of Bax/Bcl-2 indicate they may have an important role in the regulation of apoptosis of SMC.
Asunto(s)
Canal Anal/anomalías , Apoptosis/genética , Apoptosis/fisiología , Diafragma Pélvico/anomalías , Recto/anomalías , Canal Anal/embriología , Animales , Modelos Animales de Enfermedad , Regulación hacia Abajo/genética , Investigaciones con Embriones , Desarrollo Embrionario/genética , Femenino , Expresión Génica/genética , Genes bcl-2/genética , Genes bcl-2/fisiología , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Músculo Estriado/embriología , Diafragma Pélvico/embriología , Embarazo , ARN Mensajero/genética , Ratas , Recto/embriología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba/genéticaRESUMEN
Sperm DNA damage is more common in infertile men and may contribute to poor reproductive performance. However, current methods for evaluating sperm DNA integrity do not reliably predict treatment outcomes, and no treatment for abnormal DNA integrity has proven clinical value.
Asunto(s)
Fragmentación del ADN , Técnicas de Diagnóstico Molecular/métodos , Espermatozoides/metabolismo , Cromatina/química , Ensayo Cometa/métodos , Ensayo Cometa/estadística & datos numéricos , Femenino , Humanos , Etiquetado Corte-Fin in Situ/métodos , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Infertilidad Masculina/diagnóstico , Infertilidad Masculina/genética , Masculino , Inutilidad Médica , Técnicas de Diagnóstico Molecular/estadística & datos numéricos , Valor Predictivo de las Pruebas , Embarazo , Espermatozoides/citología , Resultado del TratamientoRESUMEN
Neuropathological study of a 3 1/2 year old girl with familial Leigh syndrome who also harboured a rare ATPase gene mutation disclosed extensive and unusual lesions in the cerebral cortex, despite a typical histological pattern. Early lesions in the periacqueductal grey matter of the brainstem, characterised by capillary congestion and initial regressive neuronal changes, were also observed, along with TUNEL reactive neuronal cells showing morphological signs typical of apoptosis in cortical areas with neuronal cell loss. The finding of lesions in atypical brain areas and for the first time, very early regressive neuronal phenomena, suggest that early changes in crucial brain areas may have been a cause of death. The abundance of TUNEL positive nuclei in cortical areas in the present case suggests that the apoptosis may be involved in the mechanism of neuronal death in Leigh syndrome.
Asunto(s)
Adenosina Trifosfatasas/genética , Apoptosis , Corteza Cerebral/patología , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Enfermedad de Leigh/patología , Apoptosis/genética , Apoptosis/fisiología , Tronco Encefálico/patología , Recuento de Células , Preescolar , Daño del ADN/genética , Fragmentación del ADN/genética , Fragmentación del ADN/fisiología , Femenino , Humanos , Enfermedad de Leigh/diagnóstico , Enfermedad de Leigh/genética , Mutación , Neuronas/patologíaAsunto(s)
Ensayo Cometa/métodos , Daño del ADN , Etiquetado Corte-Fin in Situ/métodos , Animales , Ensayo Cometa/instrumentación , Ensayo Cometa/estadística & datos numéricos , Humanos , Etiquetado Corte-Fin in Situ/instrumentación , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Microscopía Fluorescente/métodos , Sensibilidad y EspecificidadRESUMEN
OBJECTIVES: To investigate cell proliferation, apoptosis, and angiogenesis and their roles in transitional cell carcinoma (TCC) of the renal pelvis and ureter. METHODS: Formalin-fixed and paraffin-embedded tissue blocks from 42 patients with TCC of the renal pelvis and ureter were studied. Cell proliferation was assessed by Ki-67 immunostaining, and the proliferation index (PI) was expressed as a percentage of Ki-67-positive cells. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of TUNEL positive cells. Angiogenesis was evaluated by CD31 immunostaining, and microvessel density (MVD) was expressed as the average of the microvessel count. RESULTS: The PI ranged from 5.9% to 48.0% (median 20.03%), AI from 1.0% to 4.2% (median 2.26%), and MVD from 16.0 to 146.0 (median 56.88) in TCC of the renal pelvis and ureter. Statistical analysis revealed close associations of both PI and MVD with tumor stage and of AI with tumor grade. Our study demonstrated a strong relationship between PI and MVD, but did not show associations of AI with PI or MVD in TCC of the renal pelvis and ureter. CONCLUSIONS: It is suggested that the high activity of tumor cell proliferation with rich neovascularization may be related to the high malignant potential of the cancer, and evaluation of cell proliferation combined with angiogenesis may be useful in predicting the progression of the renal pelvic and ureteral TCC.
Asunto(s)
Carcinoma de Células Transicionales/patología , Neoplasias Renales/patología , Pelvis Renal/patología , Uréter/patología , Neoplasias Ureterales/patología , Adulto , Anciano , Apoptosis , Carcinoma de Células Transicionales/irrigación sanguínea , Carcinoma de Células Transicionales/diagnóstico , División Celular , Femenino , Humanos , Inmunoensayo/métodos , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Antígeno Ki-67 , Neoplasias Renales/irrigación sanguínea , Neoplasias Renales/diagnóstico , Pelvis Renal/irrigación sanguínea , Masculino , Persona de Mediana Edad , Neovascularización Patológica/diagnóstico , Neovascularización Patológica/patología , Uréter/irrigación sanguínea , Neoplasias Ureterales/irrigación sanguínea , Neoplasias Ureterales/diagnósticoRESUMEN
The present study investigated the frequency and distribution of TUNEL-positive neurons in brains of dementia with Lewy bodies (DLB) in comparison with those in brains of Alzheimer's disease (AD), Down syndrome (DS) and non-demented elderly persons. In DLB brains, TUNEL-positive neurons were increased in frequency compared with those in non-demented elderly brains, and showed a distribution similar to those in AD and DS brains. DLB cases with TUNEL-positive neurons showing severe Lewy pathology were all neocortical type, while DLB cases of the limbic type showing mild Lewy pathology did not demonstrate TUNEL-positive neurons. In addition, we investigated the relationships between TUNEL-positive neurons and pathological hallmarks of DLB or AD brains. TUNEL-positive neurons had no Lewy bodies or neurofibrillary tangles, and were not located within amyloid deposits. These findings suggest that neuronal damage showing DNA fragmentations occurs in DLB brains as well as in AD and DS brains, and that it is accelerated by progression of Lewy pathology as well as Alzheimer pathology, although it is not directly related to their pathological hallmarks.
Asunto(s)
Enfermedad de Alzheimer/patología , Encéfalo/patología , Etiquetado Corte-Fin in Situ , Enfermedad por Cuerpos de Lewy/patología , Neuronas/patología , Anciano , Anciano de 80 o más Años , Recuento de Células , Femenino , Humanos , Inmunohistoquímica , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Masculino , Persona de Mediana EdadRESUMEN
APP is a transmembrane precursor of beta-amyloid, and its mutations cause early-onset familial Alzheimer's disease. We report a toxic function of normal wild-type APP (wtAPP). Treatment of neuronal F11 cells, immortalized embryonic day 13 neurons, overexpressing wtAPP with anti-APP antibodies caused death. Death was not induced by antibody in parental F11 cells. Death by antibody occurred through cell-surface APP, not through secreted APP, in a pertussis toxin-sensitive manner and was typical apoptosis, not observed in primary astrocytes or glioma cells overexpressing wtAPP, but observed in primary cortical neurons. Cell-surface APP thus performs a toxic function as an extracellularly controllable regulator of neuronal death. This study provides a novel insight into the normal and pathological functions of cell-surface wtAPP.
Asunto(s)
Enfermedad de Alzheimer/inmunología , Precursor de Proteína beta-Amiloide/metabolismo , Anticuerpos/toxicidad , Apoptosis/inmunología , Línea Celular Transformada/efectos de los fármacos , Proteínas de la Membrana/metabolismo , Neurotoxinas/farmacología , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/fisiopatología , Precursor de Proteína beta-Amiloide/efectos de los fármacos , Precursor de Proteína beta-Amiloide/inmunología , Animales , Anticuerpos/inmunología , Anticuerpos/metabolismo , Especificidad de Anticuerpos/inmunología , Apoptosis/efectos de los fármacos , Recuento de Células/estadística & datos numéricos , Línea Celular Transformada/inmunología , Línea Celular Transformada/metabolismo , Inhibidores de Cisteína Proteinasa/farmacología , Feto , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Proteínas de la Membrana/efectos de los fármacos , Proteínas de la Membrana/inmunología , Neurotoxinas/inmunología , Neurotoxinas/metabolismo , Oligopéptidos/farmacología , Toxina del Pertussis , Estructura Terciaria de Proteína/efectos de los fármacos , Estructura Terciaria de Proteína/fisiología , Ratas , Factores de Virulencia de Bordetella/farmacologíaRESUMEN
This study investigated the basis for the high severity of damage to skeletal muscle due to eccentric exercise, i.e., to muscles generating force while lengthened. Fast and slow rat leg muscles maintained in an extended position were examined after 2-24 h of continuous stimulation. The treatment caused the injury to some regions of both muscles. Within the better preserved parts of the muscles, i.e., those without signs of necrotic processes, dystrophin, spectrin, and some of the dystrophin-associated proteins (beta-dystroglycan, alpha-sarcoglycan, and gamma-sarcoglycan) disappeared from sarcolemma of many fibers. The reduction or loss of dystrophin from the sarcolemma was more evident than that of other proteins examined, with sarcoglycans apparently being the most preserved. Several muscle fibers devoid of dystrophin contained apoptotic nuclei. Simultaneously, Bax, Bcl-2 and caspase-3 proteins appeared in many fibers. Our results indicate that a normal muscle overworking in an extended position undergoes the loss of several membrane skeletal proteins because of the excessive stress to the membrane cytoskeleton, which can lead to fiber death by either apoptosis or necrosis. This experimental model may represent a good model for mimicking the pathogenetic events in several muscular dystrophies.
Asunto(s)
Apoptosis/fisiología , Distrofina/metabolismo , Fatiga Muscular/fisiología , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/metabolismo , Animales , Caspasa 3 , Caspasas/metabolismo , Proteínas del Citoesqueleto/metabolismo , Distroglicanos , Estimulación Eléctrica , Femenino , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Glicoproteínas de Membrana/metabolismo , Contracción Muscular/fisiología , Fibras Musculares de Contracción Rápida/metabolismo , Fibras Musculares de Contracción Rápida/patología , Fibras Musculares de Contracción Lenta/metabolismo , Fibras Musculares de Contracción Lenta/patología , Músculo Esquelético/lesiones , Músculo Esquelético/patología , Condicionamiento Físico Animal/fisiología , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Wistar , Sarcoglicanos , Proteína X Asociada a bcl-2RESUMEN
The extent of DNA fragmentation analysed using the TUNEL technique was evaluated in post-mortem human brain tissue. Twenty-four patients with clinical and histopathological diagnosis of Alzheimer's disease (AD) and a short post-mortem delay were analysed. We report an increase in the count of TUNEL-labelled cells as the pathology of AD intensifies. Our results point out a significant correlation between neurofibrillary tangle and senile/neuritic plaque score and TUNEL-labelled cells. Patients with two copies of apolipoprotein (Apo) Eepsilon4 allele had highest number of histopathological hallmarks lesions of AD, whereas the ApoE genotype did not significantly influence the density of TUNEL-positive cells. No significant correlation was found between beta-amyloid protein load and TUNEL-labelled cells. There was no relationship between the age at death, age at onset, extent of astrogliosis or microgliosis and TUNEL-labelled cells in our material.
Asunto(s)
Enfermedad de Alzheimer/patología , Apoptosis/genética , Fragmentación del ADN/fisiología , Gliosis/patología , Degeneración Nerviosa/patología , Anciano , Anciano de 80 o más Años , Alelos , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Apolipoproteína E4 , Apolipoproteínas E/genética , Astrocitos/metabolismo , Astrocitos/patología , Encéfalo/metabolismo , Encéfalo/patología , Encéfalo/fisiopatología , Femenino , Gliosis/genética , Gliosis/metabolismo , Humanos , Etiquetado Corte-Fin in Situ/estadística & datos numéricos , Masculino , Microglía/metabolismo , Microglía/patología , Degeneración Nerviosa/genética , Degeneración Nerviosa/metabolismo , Ovillos Neurofibrilares/metabolismo , Ovillos Neurofibrilares/patología , Neuronas/metabolismo , Neuronas/patología , Placa Amiloide/metabolismo , Placa Amiloide/patologíaRESUMEN
This study reports that postischemic apoptotic cell death of the hippocampal cornu ammonis (CA) 1 neurons is delayed in aged gerbils. Age-related changes in the process of CA1 neuronal death following transient ischemia was studied. Two groups of Mongolian gerbils were used in this study, which compared adult (4-month-old) and aged (24-month-old) animals by hematoxylin-eosin stain, in situ nick-end labeling (TUNEL method) and electron microscopy. In the process of neuronal death, neuronal loss of the aged group was histologically less severe than that of the adult group. TUNEL-positive cells were found on days 3-5 after ischemia in the adult group, while they were still found on day 7 in the aged group. The apoptotic process of the aged group was delayed compared to the adult group. Furthermore, lipofuscin was ultrastructurally observed inside the apoptotic body 5 days after ischemia in CA1 pyramidal neurons of the aged group. It is likely that colocalization of lysosomal enzyme cathepsin D with lipofuscin might be associated with the age-related alteration of lysosomal system in the neurons. Altogether these data suggest that age-related lysosomal changes might affect the apoptotic cascade process in postischemic CA1 neurons.