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Mutación , Trastornos Parkinsonianos/genética , Trastornos Parkinsonianos/patología , Proteínas Serina-Treonina Quinasas/genética , Tauopatías/genética , Tauopatías/patología , Anciano , Humanos , Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina , Masculino , Trastornos Parkinsonianos/complicaciones , Tauopatías/complicaciones , Tomografía Computarizada de Emisión de Fotón Único , Temblor/etiología , Temblor/genética , Temblor/patologíaRESUMEN
OBJECTIVE: To define distinctive features of nodular heterotopia in specimens derived from drug-resistant patients with epilepsy by evaluating mRNA expression of three different layer-specific markers: Rorbeta, Er81, and Nurr1. METHODS: We analyzed the expression profile of these genes, recognized as markers mainly expressed in layer IV for Rorbeta, in layer V for Er81, and in layer VI for Nurr1, in surgical samples from 14 epileptic patients, using in situ hybridization. Six patients had subcortical nodular heterotopia and 8 patients were controls. The intrinsic organization of nodular formations and of the overlaying neocortex was assessed. RESULTS: In all patients, the 3 selected genes showed high cortical laminar specificity. In subcortical nodular heterotopia, the different gene expression profiles revealed a rudimentary laminar organization of the nodules. In the overlaying cortex, fewer cells expressed the 3 genes in the appropriate specific layer as compared to controls. CONCLUSIONS: These data provide new insights into possible ontogenetic mechanisms of nodular heterotopia formation and show the potential role of layer-specific markers to elucidate the neuropathology of malformations of cortical development.
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Corteza Cerebral/anomalías , Corteza Cerebral/patología , Heterotopia Nodular Periventricular/genética , Heterotopia Nodular Periventricular/patología , Adolescente , Adulto , Corteza Cerebral/fisiología , Niño , Proteínas de Unión al ADN/biosíntesis , Proteínas de Unión al ADN/genética , Femenino , Perfilación de la Expresión Génica/métodos , Marcadores Genéticos/genética , Humanos , Masculino , Malformaciones del Desarrollo Cortical/genética , Malformaciones del Desarrollo Cortical/patología , Miembro 2 del Grupo F de la Subfamilia 1 de Receptores Nucleares , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares , Receptores Citoplasmáticos y Nucleares/biosíntesis , Receptores Citoplasmáticos y Nucleares/genética , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética , Adulto JovenRESUMEN
BACKGROUND AND AIMS: Chronic inflammatory demyelinating polyneuropathy (CIDP) seems to be more common in patients with diabetes than in the general population. The long term outcome of these patients after receiving intravenous immunoglobulin is unclear and the precise optimal regimen needed has yet to be ascertained. Moreover, the influence of chronic hyperglycaemia on this neuropathy is not clear. METHODS: This prospective follow-up study included all consecutive patients with diabetes with a CIDP referred to our department during the 18 months of the study. RESULTS: 198 consecutive patients were referred to our neuromuscular unit and exhaustively screened. 16 patients with diabetes (8%) had a demyelinating polyneuropathy fulfilling the most restrictive diagnostic criteria for CIDP. They were treated with at least one course of intravenous immunoglobulin and, if responders, retreated in case of relapse. All patients were followed for at least 40 months. Patients with diabetes with CIDP significantly improved after immunotherapy and during follow-up. The Neuropathy Impairment Score changed from 38 at presentation to 16 at the end of the follow-up. Eight patients developed distal sensory disturbances during follow-up and four of these patients complained of distal paresthesias but no neuropathic pain. Sensory disturbances were detected after 30 months (mean time) from baseline. CONCLUSION: CIDP is not an unusual neuropathy in patients with diabetes. Our study underlines the importance of extensively investigating patients with diabetes with polyneuropathy to identify those with a treatment responsive demyelinating polyneuropathy.
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Neuropatías Diabéticas/terapia , Inmunoglobulinas Intravenosas/administración & dosificación , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/terapia , Adulto , Anciano , Biopsia , Enfermedades Desmielinizantes/patología , Neuropatías Diabéticas/inmunología , Neuropatías Diabéticas/patología , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Fibras Nerviosas Mielínicas/patología , Conducción Nerviosa , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/inmunología , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/patología , Estudios Prospectivos , Nervio Sural/patología , Resultado del TratamientoRESUMEN
Intraoperative [111In]-pentetreotide scintigraphy with a hand-held gamma detector probe has recently been proposed to increase the intraoperative detection rate of small neuroendocrine tumors and their metastases. We report a case of a 28-yr-old woman with ectopic Cushing's syndrome due to an ACTH-secreting bronchial carcinoid, in whom the use of radioguided surgery improved disease management. At presentation, radiolabeled pentetreotide scintigraphy was the only procedure able to detect the ectopic source of ACTH. After radiologic confirmation, the patient underwent removal of a bronchial carcinoid, with disease persistence. After surgery, pentetreotide scintigraphy showed pathologic uptake in the mediastinum not previously detected at surgery and only subsequently confirmed by radiologic studies. Despite a second thoracic exploration, hormonal, scintigraphic, and radiological evidence of residual disease persisted. Radioguided surgery was then performed using a hand-held gamma probe 48 h after iv administration of a tracer dose of radiolabeled [111In-DTPA-D-Phe1]-pentetreotide, which permitted detection and removal of multiple residual mediastinal lymph node metastases. Clinical and radiologic cure, with no evidence of tracer uptake at pentetreotide scintigraphy, was subsequently observed. The use of an intraoperative gamma counter appears a promising procedure in the management of metastatic ACTH-secreting bronchial carcinoids.
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Síndrome de ACTH Ectópico/diagnóstico por imagen , Síndrome de ACTH Ectópico/cirugía , Neoplasias de los Bronquios/diagnóstico por imagen , Neoplasias de los Bronquios/cirugía , Tumor Carcinoide/diagnóstico por imagen , Tumor Carcinoide/cirugía , Síndrome de Cushing/etiología , Radiofármacos , Somatostatina/análogos & derivados , Cirugía Asistida por Computador , Síndrome de ACTH Ectópico/etiología , Hormona Adrenocorticotrópica/sangre , Adulto , Biomarcadores , Neoplasias de los Bronquios/complicaciones , Tumor Carcinoide/complicaciones , Hormona Liberadora de Corticotropina , Desamino Arginina Vasopresina , Femenino , Hormonas/sangre , Humanos , Hidrocortisona/sangre , Mediciones Luminiscentes , Metástasis Linfática , Neoplasias del Mediastino/cirugía , Octreótido , Cintigrafía , Estimulación Química , Tomografía Computarizada por Rayos XRESUMEN
We present the results of a retrospective study on 10 patients operated on for intractable epilepsy associated with nodular heterotopia as identified by high resolution MRI. Seven patients had unilateral heterotopia, one patient had symmetric bilateral heterotopia and two patients had asymmetric bilateral heterotopia. By stereo-electroencephalogram (SEEG) (nine patients) interictal activity within nodules was similar in all cases, and ictal activity never started from nodules alone but from the overlying cortex or simultaneously in nodules and cortex. Excellent outcomes (Engel class Ia, 1987) were achieved in the seven patients with unilateral heterotopia, showing that surgery can be highly beneficial in such cases when the epileptogenic zone is carefully located prior to surgery by MRI and particularly SEEG. For the bilateral cases surgical outcomes were Engel IIa (one patient) or Engel IIIa (two patients). Histological/immunohistochemical studies of resected specimens showed that all nodules had similar microscopic organization, even though their extent and location varied markedly. The overlying cortex was dysplastic in nine patients, but of normal thickness. We suggest that nodule formation may be the result of a dual mechanism: (i) failure of a stop signal in the germinal periventricular region leading to cell overproduction; and (ii) early transformation of radial glial cells into astrocytes resulting in defective neuronal migration. The intrinsic interictal epileptiform activity of nodules may be due to an impaired intranodular GABAergic system.
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Encefalopatías/cirugía , Corteza Cerebral , Coristoma/cirugía , Epilepsia/cirugía , Adulto , Encefalopatías/complicaciones , Encefalopatías/patología , Encefalopatías/fisiopatología , Corteza Cerebral/patología , Coristoma/complicaciones , Coristoma/patología , Coristoma/fisiopatología , Electroencefalografía , Epilepsia/etiología , Epilepsia/patología , Epilepsia/fisiopatología , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
AIM: The outcome of surgery in gastric cancer differs in Japan and Western countries and the extension of lymphadenectomy may play a crucial role in survival. In Japan the choice of performing extended (D2) and superextended (D4) lymphadenectomies is based on retrospective studies, and a prospective randomized study comparing D2 and D4 is still in course. In Western countries the randomized trials comparing D1 and D2 could not provide definite indications, D2 is not yet performed as a routine procedure and D4 is accepted only by few surgeons. We report our experience and discuss indications and results. METHODS: Since January 2000 through December 2002 we performed 27 superextended lymphadenectomies for the radical treatment of advanced gastric cancer. Early gastric cancers and patients over 80 years of age received conventional D2 gastrectomies. Selection of patients for D4 was made after laparotomy, when intraoperative peritoneal lavage cytology could rule out the presence of malignant cells, while D2 was done in case of peritoneal micrometastases. RESULTS: Every patients had 39.5 nodes removed on average (range 17-94), and micrometastases in tier 16 were found in 7 cases (26%). Early post-operative surgical morbidity was 18% (5 patients) and mortality was 3.7% (1 patient). As much as 30% of patients complained of diarrhea as a late complication. The follow up could demonstrate a 3 year overall actuarial survival of 76%. Actuarial survival was 100% for N- and 70% for N+. A remarkable data was that 4 out of 5 patients who died from recurrence in the follow-up, were N4+. Actuarial survival at 3 years for N4+ patients was 34%, and the difference in survival between N4+ and other N+ was statistically significant (p<0.05). CONCLUSIONS: Superextended lymphadenectomy in gastric cancer is feasible with postoperative morbidity and mortality rates not exceeding the rates of other lymphadenectomies. Actuarial survival at 3 years with D4 was better than in previous personal experience with D2, although the patients who underwent D4 were selected by intraperitoneal lavage cytology, while D2 patients had not been selected. The prognosis for N4- patients was better than for N4+ with micrometastases in tier 16. The presence of N4 micrometastases worsens the prognosis, but it is still uncertain whether D4 does improve survival: it is undoubtedly a new means of more accurate staging in gastric cancer surgery. The newer TNM classification regards the number of nodes removed as an indicator of radicality. Every surgeon should consider that superextended lymphadenectomies could comply with R0 radicality, and perform it within the ranges of low morbidity and mortality, until randomized trials with definitive results are available.
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Adenocarcinoma/cirugía , Escisión del Ganglio Linfático/métodos , Metástasis Linfática , Neoplasias Gástricas/cirugía , Análisis Actuarial , Adenocarcinoma/mortalidad , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Factibilidad , Femenino , Estudios de Seguimiento , Gastrectomía , Humanos , Masculino , Persona de Mediana Edad , Selección de Paciente , Complicaciones Posoperatorias , Pronóstico , Neoplasias Gástricas/mortalidad , Factores de TiempoRESUMEN
Patients with diabetes mellitus (DM) may develop chronic inflammatory demyelinating polyneuropathy (CIDP), which may be difficult to distinguish from diabetic neuropathy (DNP). Here the authors show that immunoreactivity for matrix metalloproteinase-9 on sural nerve biopsies may help to identify CIDP-DM. In a pilot study on 10 CIDP-DM patients with IV immunoglobulins and tight glycemic control, the CIDP-DM patients had a better outcome than DNP patients treated with tight glycemic control only.
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Complicaciones de la Diabetes , Metaloproteinasa 9 de la Matriz/análisis , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/diagnóstico , Nervio Sural/enzimología , Adulto , Anciano , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/tratamiento farmacológico , Neuropatías Diabéticas/diagnóstico , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Humanos , Inmunohistoquímica , Masculino , Metaloproteinasa 9 de la Matriz/inmunología , Persona de Mediana Edad , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/complicaciones , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/tratamiento farmacológico , Nervio Sural/patologíaRESUMEN
Since the original description by Taylor, the term focal cortical dysplasia has been used to refer to a wide range of alterations of the cortical mantle. More recently, these conditions have been described from neuroimaging, neuropathological and genetic standpoints, generating several classifications. It is widely recognized that these classifications are unsatisfactory. We propose a simplified classification of focal cortical dysplasias based on easily recognized neuropathological characteristics. We retrospectively re-examined histological sections of cortex from 52 of 224 (23%) patients operated on for drug-resistant partial epilepsy in which cortical dysplasia was present but not associated with other brain pathologies except hippocampal sclerosis. Three subgroups were identified: (i) architectural dysplasia (31 patients) characterized by abnormal cortical lamination and ectopic neurones in white matter; (ii) cytoarchitectural dysplasia (six patients) characterized by giant neurofilament-enriched neurones in addition to altered cortical lamination; and (iii) Taylor-type cortical dysplasia (15 patients) with giant dysmorphic neurones and balloon cells (all but two patients) associated with cortical laminar disruption. The patients with architectural dysplasia had lower seizure frequency than those with cytoarchitectural and Taylor-type dysplasia, and the epileptogenic zone was mainly in the temporal lobe. In patients with Taylor-type dysplasia, the epileptogenic zone was mainly extratemporal, and interictal stereo-EEG was distinctive. MRI was unrevealing in 34% of patients, but distinctive signal alterations characterized most patients with Taylor-type dysplasia, while focal hypoplasia with MRI abnormalities was found in architectural dysplasia. Patients with Taylor-type dysplasia had the best outcome, with 75% seizure-free (Engel class Ia) after at least a year of follow-up compared with 50% of cytoarchitectural dysplasia and 43% of architectural dysplasia patients seizure-free. This three-category classification is based on easily recognized histopathological characteristics and avoids complicated terminology, while the distinctive ensemble of other characteristics defines clinically homogeneous groups.
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Corteza Cerebral/anomalías , Corteza Cerebral/cirugía , Electroencefalografía , Epilepsia/cirugía , Adulto , Edad de Inicio , Corteza Cerebral/patología , Epilepsia/clasificación , Epilepsia/patología , Estudios de Seguimiento , Humanos , Imagen por Resonancia Magnética , Persona de Mediana Edad , Neuronas/patología , Estudios Retrospectivos , Factores de Tiempo , Resultado del TratamientoRESUMEN
Forty consecutive patients with chronic sensorimotor polyneuropathy of undetermined cause were followed to determine disease progression and prognosis. They had had neurological symptoms for at least 1 year before presentation. Neurophysiological examination in all patients showed chronic axonal degeneration, which was confirmed by sural nerve biopsy. Patients were reviewed every 3 months. Laboratory tests were performed every 6 months for the first 2 years and then annually. Neurophysiological examination was performed annually in all patients. Patients were followed up for at least 4 years. In no instance was a possible etiological factor detected during follow-up. The clinical and electrophysiological findings had a slowly progressive course. We suggest that patients with a chronic polyneuropathy of undetermined cause despite detailed investigations do not require further extensive and expensive laboratory tests and neurophysiological studies during follow-up.
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Polineuropatías/etiología , Polineuropatías/patología , Anciano , Anciano de 80 o más Años , Evaluación de la Discapacidad , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Masculino , Nervio Mediano/patología , Persona de Mediana Edad , Conducción Nerviosa , Dimensión del Dolor , Nervio Peroneo/patología , Polineuropatías/terapia , Estudios Prospectivos , Nervio Sural/patologíaRESUMEN
Cajal-Retzius cells, identified using calretinin antiserum, were studied in layer I (LI) of adult human temporal cortex from epileptic patients with Taylor's focal cortical dysplasia and architectural dysplasia, in comparison with normal cortex. Both types of dysplasia showed LI hypercellularity, but only in architectural dysplasia was the density of Cajal-Retzius cells significantly increased. A subset of Cajal-Retzius cells were reelin immunoreactive, but none were GABA positive. These findings suggest that differences in the persistence of Cajal-Retzius cells, which probably reflect different types of alteration during brain development, can assist in characterizing different forms of cortical dysplasia.
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Epilepsia/patología , Malformaciones del Sistema Nervioso/patología , Neuronas/clasificación , Neuronas/patología , Lóbulo Temporal/patología , Moléculas de Adhesión Celular Neuronal/biosíntesis , Recuento de Células , Epilepsia/complicaciones , Proteínas de la Matriz Extracelular/biosíntesis , Técnica del Anticuerpo Fluorescente , Humanos , Microscopía Confocal , Proteínas del Tejido Nervioso , Malformaciones del Sistema Nervioso/complicaciones , Neuronas/metabolismo , Proteína Reelina , Serina EndopeptidasasRESUMEN
PURPOSE: Different types of epilepsies and seizures depend on the nature and location of the primary disturbance and are presumably mediated by different physiopathological mechanisms. We immunocytochemically investigated possible changes in the inhibitory-aminobutyric acid (GABA)ergic system in specimens taken from four patients who underwent surgery for intractable epilepsy and presented two different types of focal cortical dysplasia in the temporal lobe. METHODS: The patients were selected on the basis of electroclinical, imaging, and routine neuropathological data: two had Taylor focal dysplasia, and two had non-Taylor dysplasia (microdysgenesia). The study was performed using antibodies against parvalbumin (PV), glutamic acid decarboxylase (GAD), and GABA-transporter 1 (GAT1). RESULTS: In the patients with Taylor dysplasia, laminar disorganization of the cortex was associated with the presence of giant neurons and ballooned cells; there was a reduced number of PV-positive neurons and terminals, the giant neurons were surrounded by clusters of PV- and GAD-positive terminals, and there was an overall reduction in GAT1. Despite the presence of cortical laminar disorganization, no giant or ballooned cells were found in the patients with non-Taylor microdysgenesia; there was a marked decrease in PV and GAD immunoreactive elements, with a patchy distribution of GAD and GAT1 immunoreactivity but no clustering of PV and GAD terminals. CONCLUSIONS: These results suggest that the two forms of cortical dysplasia are characterized by different and selective morphofunctional alterations in the GABAergic system.
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Corteza Cerebral/anomalías , Corteza Cerebral/patología , Epilepsia del Lóbulo Temporal/patología , Inhibición Neural/fisiología , Vías Nerviosas/patología , Neuronas/patología , Lóbulo Temporal/patología , Adulto , Corteza Cerebral/fisiopatología , Epilepsia del Lóbulo Temporal/fisiopatología , Femenino , Humanos , Inmunohistoquímica , Imagen por Resonancia Magnética , Masculino , Lóbulo Temporal/fisiopatología , Ácido gamma-Aminobutírico/fisiologíaRESUMEN
In the present report we describe the neuropathological characteristics of tissue surgically resected from three patients affected by intractable epilepsy secondary to cortical dysplasia. Common features, suggestive of a focal cortical dysplasia of Taylor, were observed in all specimens. Immunocytochemical procedures were performed using neuronal and glial markers and the sections were observed at light traditional and confocal microscopes. This part of the investigation pointed out: 1. cortical laminar disruption; 2. very large neurons displaying a pyramidal or round shape; 3. ballooned cells; 4. decrease of calcium binding proteins immunoreactivity; 5. abnormal nets of parvalbumin- and glutamic acid decarboxylase-positive puncta around giant neurons but not around ballooned cells. Ultrastructural investigation on the same material provided evidence of a high concentration of neurofilaments in giant neurons and of glial intermediate filaments in ballooned cells. In addition, immunolabeled GABAergic terminals clustered around giant neurons were not found to establish synapses on their cell bodies. The present data, derived from a limited sample of patients but showing very consistent features, suggest that in Taylor's type of cortical dysplasia a disturbance of migratory events could be paralleled by a disruption of cell differentiation and maturation and by an impairment of synaptogenesis. This latter mechanism seemed to affect especially the inhibitory elements, and could account for the hyperexcitability of this tissue and thus for the high epileptogenicity of Taylor's dysplasia.
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Corteza Cerebral/anomalías , Corteza Cerebral/ultraestructura , Adulto , Calbindina 2 , Calbindinas , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Niño , Epilepsia/etiología , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Glutamato Descarboxilasa/metabolismo , Humanos , Inmunohistoquímica , Filamentos Intermedios/ultraestructura , Masculino , Microscopía Confocal , Microscopía Electrónica , Neurofibrillas/ultraestructura , Neuronas/metabolismo , Neuronas/patología , Neuronas/ultraestructura , Parvalbúminas/metabolismo , Terminales Presinápticos/ultraestructura , Proteína G de Unión al Calcio S100/metabolismo , Vimentina/metabolismoRESUMEN
AIMS AND BACKGROUND: The aim of this study was to compare the local immune response in two groups of patients with high-grade cervical intraepithelial squamous lesions (SIL): one with HIV infection and the other with HPV infection alone. MATERIALS AND METHODS: 16 conization specimens (8 from HIV-infected and 8 from non-HIV-infected patients) of HPV-related, high-grade SIL were selected. The specimens from non-HIV patients were considered as controls. The total number of Langerhans cells, CD4 and CD8 cells present in 10 field areas (3.120 mm2) was recorded in each case. In HIV patients CD4 and CD8 peripheral counts were performed immediately before surgery. RESULTS: The CD4/CD8 ratio never exceeded 0.71, whereas the lowest ratio in controls was 0.81: this difference was statistically significant (P = 0.0009). The mean number of Langerhans cells was markedly reduced in the high-grade SILs in the HIV patients in comparison with controls (P = 0.001). The number of CD4 cells and the CD4/CD8 ratio correlated with the peripheral CD4 count (P = 0.001 and 0.02). CONCLUSIONS: In our study a marked local impairment of cervical immunoreactivity was observed, which may play a major role in the progression of these lesions in HIV-infected women.
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Infecciones por VIH/inmunología , Papillomaviridae , Infecciones por Papillomavirus/inmunología , Infecciones Tumorales por Virus/inmunología , Displasia del Cuello del Útero/inmunología , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/inmunología , Neoplasias del Cuello Uterino/virología , Adulto , Relación CD4-CD8 , ADN Viral/análisis , Femenino , Infecciones por VIH/complicaciones , Humanos , Papillomaviridae/genética , Infecciones por Papillomavirus/complicaciones , Infecciones Tumorales por Virus/complicacionesRESUMEN
OBJECTIVE: In patients with Cushing's disease multihormonal responses to ovine corticotrophin releasing hormone (oCRH) have been detected in blood from inferior petrosal sinuses. This finding has been explained by co-secretion of other hormones, in addition to ACTH, by the pituitary adenoma itself or by paracrine effects exerted by the adenoma on normal periadenomatous pituitary cells. To assess these hypotheses we compared the presence of a CRH induced GH and/or PRL response during inferior petrosal sinus sampling to the immunohistochemical detection of PRL and GH in adenomatous tissue removed from patients with Cushing's disease. PATIENTS AND MEASUREMENTS: Twenty-two patients with Cushing's disease and two patients with ectopic ACTH syndrome due to a bronchial carcinoid were studied; each patient had undergone preoperative inferior petrosal sinus sampling for diagnostic purposes with determination of GH and PRL in addition to ACTH, before and after administration of oCRH. Immunohistochemical studies for ACTH, GH and PRL detection were carried out on adenomatous tissue removed at surgery in the patients with pituitary dependent Cushing's disease and on the carcinoid tumours from the two patients with ectopic ACTH syndrome. RESULTS: All pituitary adenomas immunostained for ACTH, and four adenomas immunostained for GH or PRL in addition to ACTH. A PRL increase in the inferior petrosal sinus after oCRH administration was found in 11 of 22 patients, but none of their tumours immunostained for PRL. Immunostaining for PRL was found in the pituitary tumours from two patients but in neither patient was there a PRL response after oCRH. A GH response was found in 13 of 20 patients in whom it was sought; no patient showed immunostaining in their tumour. GH immunostaining was found in two tumours but in neither patient was there a GH response after oCRH. The oCRH-induced increase of GH and PRL was always recorded in the dominant inferior petrosal sinus. The ACTH response to oCRH was significantly higher in patients who showed oCRH induced increases in GH and/or PRL than in patients who did not, both in terms of area under the response-curve (22,032 +/- 9876 vs. 4371 +/- 2870 ng/l/10 min; P < 0.05) or mean percentage increase above baseline (754 +/- 229% vs. 147.2 +/- 67%, P < 0.02). A significant correlation was observed between ACTH and GH responses to oCRH. The two patients with ectopic Cushing's syndrome did not show ACTH, GH or PRL increases after oCRH administration and did not show immunostaining for GH or PRL in their tumours. CONCLUSIONS: The present data do not support the hypothesis of co-secretion of hormones by the pituitary adenoma as the cause of the GH and PRL responses to ovine corticotrophin releasing hormone observed in patients with Cushing's disease; it is suggested that a different mechanism, possibly involving an interaction between the ACTH secreting adenoma and the normal periadenomatous GH and PRL secreting cells, may be responsible.
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Hormona Liberadora de Corticotropina , Síndrome de Cushing/diagnóstico , Hormona del Crecimiento/metabolismo , Comunicación Paracrina , Prolactina/metabolismo , Adenoma/química , Adenoma/complicaciones , Adenoma/metabolismo , Adolescente , Hormona Adrenocorticotrópica/análisis , Hormona Adrenocorticotrópica/sangre , Hormona Adrenocorticotrópica/metabolismo , Adulto , Anciano , Síndrome de Cushing/sangre , Síndrome de Cushing/etiología , Femenino , Hormona del Crecimiento/análisis , Hormona del Crecimiento/sangre , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Muestreo de Seno Petroso , Neoplasias Hipofisarias/química , Neoplasias Hipofisarias/complicaciones , Neoplasias Hipofisarias/metabolismo , Prolactina/análisis , Prolactina/sangre , Estudios RetrospectivosRESUMEN
An involvement of mycobacteria in the pathogenesis of sarcoidosis has often been hypothesized, but not confirmed reproducibly. In this study we applied a nested Polymerase Chain Reaction (PCR) to the insertion sequence IS6110 for detection of Mycobacterium tuberculosis (MT) DNA in formalin-fixed and paraffin-embedded tissues from patients with sarcoidosis, with tuberculosis and atypical mycobacteriosis confirmed by culture, and from negative control samples. MT-DNA could be detected in 2/30 samples of sarcoidosis, in 10/10 tuberculoses, in 0/5 atypical mycobacterioses and in 0/10 negative controls. Nested PCR confirmed its high sensitivity and specificity in detecting MT-DNA on archival histopathological specimens. From our results we conclude that in the granulomatous lesions of sarcoidosis MT-DNA is only sporadically demonstrable and probably it doesn't play a role in the pathogenesis of this disease.
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ADN Bacteriano/análisis , Ganglios Linfáticos/microbiología , Mycobacterium tuberculosis/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Sarcoidosis/microbiología , Fijadores , Formaldehído , Humanos , Ganglios Linfáticos/patología , Infección por Mycobacterium avium-intracellulare/complicaciones , Infección por Mycobacterium avium-intracellulare/microbiología , Infección por Mycobacterium avium-intracellulare/patología , Mycobacterium tuberculosis/genética , Adhesión en Parafina , Sarcoidosis/complicaciones , Sarcoidosis/patología , Sensibilidad y Especificidad , Fijación del Tejido , Tuberculosis Ganglionar/complicaciones , Tuberculosis Ganglionar/microbiología , Tuberculosis Ganglionar/patologíaRESUMEN
From November 1985 to December 1990, 2,552 endomyocardial biopsy specimens from 209 heart transplant patients were studied. Forty-four (21%) patients developed 45 episodes of major human cytomegalovirus infection (HCMV). Human cytomegalovirus infection was primary in 13 of 44 patients. Thirty-one patients developed episodes of recurrent major infection. One patient had both primary and recurrent infections. Conventional histopathologic and immunohistochemical study, in situ hybridization, and polymerase chain reaction were used to diagnose HCMV myocardial involvement on corresponding endomyocardial biopsy specimens performed during infection. Conventional morphologic study showed typical viral inclusion bodies in four biopsy specimens. Two cases had myocyte HCMV localization with necrotizing myocarditis, whereas two had endothelial cell involvement without any inflammatory reaction. In these four biopsy specimens, immunohistochemistry showed a higher number of infected cells than that recognized by conventional histopathologic study. In situ hybridization detected infected cells with no evidence of cytopathic effect. Polymerase chain reaction gave HCMV amplification products in two additional biopsy specimens otherwise interpreted as moderate and mild rejection, respectively. Therefore, 6 biopsies showed HCMV myocardial involvement (6 of 45; 13.3%): all were from patients with primary HCMV infection (6 of 13; 46%). None of 32 major recurrent infections showed any myocardial involvement. In conclusion, our study is the first to demonstrate that myocardial HCMV involvement preferentially occurs in primary infection and HCMV endothelial localization can be free from inflammatory reaction, whereas HCMV myocyte localization leads to necrotizing myocarditis. Polymerase chain reaction has a higher diagnostic sensitivity than in situ hybridization. However, polymerase chain reaction findings of HCMV DNA on otherwise negative endomyocardial biopsy specimens remains of questionable significance because polymerase chain reaction-positive biopsy samples do not necessarily indicate tissue infection. It is impossible to determine whether amplified sequences derive from circulating leukocytes or from tissue cells.
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Infecciones por Citomegalovirus/etiología , Trasplante de Corazón , Complicaciones Posoperatorias , Antígenos Virales/análisis , Secuencia de Bases , Biopsia , Citomegalovirus/inmunología , Citomegalovirus/aislamiento & purificación , Infecciones por Citomegalovirus/genética , Infecciones por Citomegalovirus/inmunología , Endocardio/microbiología , Endocardio/patología , Rechazo de Injerto , Corazón/microbiología , Humanos , Inmunohistoquímica , Sondas Moleculares/genética , Datos de Secuencia Molecular , Miocardio/patología , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , RecurrenciaRESUMEN
A polymerase chain reaction (PCR) amplification assay was developed to detect Coxsackievirus B3 ribonucleic acid (RNA) in blood and myocardial tissue of explanted hearts from 40 patients who underwent cardiac transplantation and in 1 normal heart. Twenty-one patients were affected by idiopathic dilated cardiomyopathy of different duration and 19 by coronary artery disease. Coxsackievirus B3 in vitro infected Vero cells and cells infected by related human enteroviruses (Coxsackievirus B2, B4, and poliovirus 1) were used as reaction controls. PCR was performed using 4 pairs of primers homologous to Coxsackie-virus B3 sequences. Three sets were located in regions of the genome conserved at nucleotide level between several enterovirus species (replicase gene, 5' noncoding region), while one was located in a Coxsackievirus B3-specific region (VP1 gene). Total RNA was prepared by acid guanidinium isothiocyanate extraction from tissue stored frozen at -80 degrees C. One microgram of total RNA was retrotranscribed with either antisense primer or with random hexanucleotide primers and then subjected to 40 cycles of amplification. PCR products were separated by electrophoresis on a 10% polyacrylamide gel, electrotransferred to a nylon membrane and then hybridized to oligonucleotide probes specific for the coxsackievirus B3 genome radiolabeled with radioactive isotope of phosphorous. All pairs of primers yielded specific amplification products when tested on Coxsackievirus B3-infected Vero cells, with a sensitivity of 1 infected cell out of 10(5) to 10(6) cells starting from 1 microgram total RNA. Primer sets for regions of Coxsackievirus B3 genome highly conserved between related enteroviral species gave positive amplification also when challenged with RNA from cells infected by Coxsackievirus B2, B4 and poliovirus 1.
Asunto(s)
Cardiomiopatía Dilatada/microbiología , Enterovirus Humano B/genética , Trasplante de Corazón , ARN Viral/análisis , Adulto , Secuencia de Bases , Southern Blotting , Cardiomiopatía Dilatada/cirugía , Enterovirus Humano B/aislamiento & purificación , Femenino , Corazón/microbiología , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos , Reacción en Cadena de la Polimerasa , ARN Viral/sangreAsunto(s)
Ferritinas/análisis , Hemocromatosis/metabolismo , Hemosiderosis/metabolismo , Miocardio/química , Adolescente , Adulto , Femenino , Humanos , Inmunohistoquímica , MasculinoRESUMEN
The authors performed an immunohistochemical study on expression of tumor necrosis factor alpha (TNF alpha) in endomyocardial biopsies from human cardiac allografts. TNF alpha immunoreactivity was found in 45% biopsies with mild acute rejection, in 83% biopsies with focal moderate rejection, in 80% biopsies with diffuse moderate rejection. Biopsies with absent rejection did not show immunoreactive cells. In mild rejection, positive cells were few and scanty monocytes and macrophages (MAC-387 and LN5 positive cells) and T lymphocytes (UCHL-1/CD45 RO positive cells) (up to 20% of all infiltrating cells). Expression of major histocompatibility complex (MHC) class II antigens on infiltrating and endothelial cells occurred earlier and independent of TNF alpha reactivity. Number of immunoreactive cells increased in moderate rejection (up to 50%). Immunoreactivity was also present in nonpigmented macrophages in part of the biopsies with resolving rejection (45%). The authors conclude that TNF alpha is expressed in acute cardiac rejection by immunologically activated inflammatory cells. Immunoreactive cells increase in number with increasing severity of the reaction.