Glutamate release and synapsin-I phosphorylation induced by P2X7 receptors activation in cerebellar granule neurons.

The present work reports that activation of P2X7 receptor induces synaptic vesicle release in granule neurons and phosphorylation of synapsin-I by calcium-calmodulin-dependent protein kinase II (CaMKII), which in turn modulates secretory event. ATP, in absence of magnesium, induced a concentration-dependent glutamate release with an EC50 value of 1.95 microM. The involvement of P2X7 receptor was suggested when maximal secretory response was significantly reduced by the selective P2X7 antagonist Brilliant Blue G (BBG; 100 nM) and abolished by removing extracellular Ca2+. The involvement of P2X7 receptor on synaptic vesicle release was confirmed by measuring the release of FM 1-43 dye. In this case, pharmacological activation of P2X7 was achieved with the more selective agonist 2'-3'-o-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP; 100 microM) showing a significant FM 1-43 release that was blocked by BBG (100 nM), by Zn2+ ions (100 microM), both P2X7 blockers, but not by suramin (100 microM), antagonist of P2X1, P2X2, P2X3 and P2X5. In addition, BzATP, through P2X7 receptor activation, significantly increased the phosphorylation of synapsin-I, the main presynaptic target of CaMKII. Both effects mediated by BzATP were inhibited by the CaMKII inhibitors KN-62 (10 microM) and KN-93 (10 microM). These results suggest, therefore, that Ca2+ entrance mediated by P2X7 receptor induces glutamate release and in parallel synapsin-I phosphorylation.

Myelodysplastic syndrome and acute nonlymphocytic leukemia secondary to mitolactol treatment in patients with breast cancer.

One thousand four hundred sixty patients with 2,590 patient-years of follow-up were treated on 15 protocols for metastatic breast cancer with dibromodulcitol (mitolactol; DBD)-containing regimens since 1976. Twenty-three patients developed myelodysplastic syndrome (MDS) and/or acute nonlymphocytic leukemia (ANLL). The overall risk of developing MDS or ANLL per person is 1.6%. In patients who had received more than 16,000 mg of DBD the risk per person is 6%, and in the high-dose subsets of patients who received no prior radiation or alkylator therapy, it is 7.9%. The risk per person increases to a maximum by 30 to 36 months (5.3%). The high risk was seen despite a study population of metastatic breast cancer patients with a median survival of 16 months. This analysis strongly suggests that DBD is one of the most potent of the reported leukemogenic-inducing agents. Further use of this drug in both the adjuvant and metastatic situation should be reconsidered.

Trisomy 5 in Ph+ chronic myeloid leukemia in association with megakaryocytosis.

A patient with a megakaryocytosis associated with a Philadelphia chromosome-positive chronic myeloid leukemia (CML) was found to have a trisomy of chromosome five. To our knowledge, this is the first case of trisomy 5 associated with a Ph + CML, particularly one with a megakaryocytosis. The trisomy 5 may be associated with the resistance to cytostatic drugs found in this patient.

Oral idarubicin in combination with cytosine-arabinoside in previously untreated patients with acute non-lymphocytic leukemia.

Twenty-six previously untreated patients with acute non-lymphocytic leukemia (ANLL) were treated with oral idarubicin and cytosine-arabinoside (Ara-C). The median age of the patients was 44 years (range, 11-72). In 23 of the 26 patients a hypoplastic marrow, with a peripheral white cell count of less than 1,000/mm3 after treatment, was documented. Treatment was well tolerated with minimal symptoms of nausea and vomiting. Diarrhea was observed in three patients and stomatitis in nine patients. Alopecia was documented in only six patients. A complete remission (CR) was obtained in 12 patients (median duration 25 weeks). The median time to CR was 3.4 weeks (range, 1.4-5). Ten of the 26 patients were alive 6 months after the start of induction treatment, while a further four patients who were in the study for less than 6 months are alive and in remission at 5, 4, 3, and 3 months, respectively. Eight of 12 patients in whom bone marrow aplasia was documented achieved a CR; perhaps the drug dosages used in this study were suboptimal.

[Bovine botulism: laboratory confirmation of clinical diagnosis in 1986-1989]. / Botulismo bovino: comprovação laboratorial do diagnóstico clínico, período 1986-1989.

Five hundred and seventeen specimens relating to a total of 235 clinical cases of bovine botulism occurring over period of four years (1986-1989) were studied. These samples came from the States of S. Paulo, Mato Grosso, Mato Grosso do Sul, Minas Gerais, Goiás, Bahia, Pará, Maranhão, Distrito Federal and Roraima. Laboratorial tests were performed on these samples with a view to toxin detection. Toxin types C and D were detected being found in the liver and in gastric and intestinal contents. The small number of confirmed cases led to the supposition of the participation of other diseases in these deaths reported, even if false negative cases are taken into account.

[Gastric neoplasm associated with IgA deficiency. Importance of multidisciplinary care]. / Neoplasia gástrica associada a défice de IgA. Importância da Multidisciplinaridade de Cuidados.

We present the case of 55-year old female patient with a rare malignant disease of the stomach associated with IgA deficiency. For its management the surgical, transfusional and renal support strategy required good coordination of pre, intra and post-operative care, and good articulation among the various sub-specialties. The patient experienced two severe anaphylactoid transfunctional reactions. Congenital IgA deficiency was detected with probable anti-IgA immunization associated with a metacrone adenocarcinoma of the stomach and nephrotic syndrome with renal failure. The transfusion methods utilized are discussed, and the results of the study aimed at detecting a compatible donor, with identical deficit, among family members are presented. In the absence of a national registry of donors with IgA deficiency, the prevention of these potentially dangerous reactions imposes a burden on the Service of Immuno-Hematherapy to take special measures on its own. This case points out the need to implement a blood bank with rare groups and IgA deficient blood, for homo/autologous transfusions.

Leukaemia in Hodgkin's disease.

Three new cases of Hodgkin's disease terminating in leukaemia are described and the literature is briefly reviewed. The patients developed acute lymphocytic leukaemia, acute eosinophilic leukaemia and chronic myeloid leukaemia, 6 1/4 years, 6 years and 7 months respectively after the initial diagnosis of Hodgkin's disease. The 2 patients who developed acute leukaemia had been in complete remission from the Hodgkin's disease and had received no treatment for 5 3/4 and 2 1/2 years respectively. Possible causes for the occurrence of leukaemia in patients with Hodgkin's disease are discussed.

Acute leukemia as a complication of long-term treatment of advanced breast cancer.

Acute nonlymphocytic leukemia occurred in four women after long-term therapy for advanced breast cancer. The patients had inoperable/metastatic breast cancer that had been well-controlled with chemotherapy for many years (7 years and 4 months, 7 years and 1 month, 6 years and 3 months, and 4 years and 7 months, respectively). Two of the patients had also received extensive radiotherapy. The chemotherapeutic agent that all four patients had received in common was cyclophosphamide.

Adjuvant chemotherapy for operable breast cancer in premenopausal women.

Surgery alone does not cure breast cancer, and adjuvant chemotherapy has changed the management of this disease. Data obtained in 81 premenopausal women with operable breast cancer, treated at our clinic, are presented. Patients with axillary node disease were treated on three different protocols: cyclophosphamide + methotrexate + fluoro-uracil + vincristine + prednisone (CMFVP), cyclophosphamide + methotrexate + fluoro-uracil (CMF), and CMF + immunotherapy with methanol extract residue of BCG (CMF + MER). Patient discriminants and treatment regimens are discussed. Analysis of the results obtained in 49 patients in one study showed an extension of disease-free survival to 4,25 years, that CMFVP was superior to CMF with or without MER, and that immunotherapy was not beneficial. The literature is briefly reviewed and the motivation for our newer studies stated.

Adjuvant chemotherapy for operable breast cancer in postmenopausal women.

Survival figures of women with operable breast cancer reached a plateau 40 years ago. It was only with the advent of adjuvant chemotherapy that these figures showed a dramatic improvement. Data obtained in 105 postmenopausal women with operable breast cancer, treated at our clinic, are presented. Patients with axillary node disease were treated on four different protocols: cyclophosphamide + methotrexate + fluoro-uracil (CMF) with or without immunotherapy was compared with CMF + vincristine + prednisone, while other studies compared observation only with CMF + prednisone and CMF + prednisone + tamoxifen. Patients older than 65 years received tamoxifen or placebo. Patient discriminants and treatment regimens are discussed. Results indicating that certain subsets of postmenopausal women definitely need adjuvant chemotherapy are presented. The literature is briefly reviewed and the motivation for our new studies explained.

Carcinoembryonic antigen in patients with breast cancer: an adjunctive tool to monitor response and therapy.

Plasma carcinoembryonic antigen (CEA) levels were performed by radioimmunoassay in 234 patients with histologically proved breast cancer: 181 with advanced metastatic disease and 53 without distant metastases but nodal involvement at time of mastectomy. Four hundred and thirty-four assays were done and correlated with the clinical status of the patients. Values above 2.5 ng/ml were taken as abnormal. Active disease was associated with elevated plasma CEA levels. Very high values were not recorded in 109 patients when they were considered to be in complete remission, while only 22 patients out of 63 patients with progressive disease had normal values. In 16 of these values remained normal despite progression of disease. In 6 patients clinical relapse preceded CEA elevation by 2--5 months. Tumor burden and abnormal serial CEA values showed positive correlation in 38 patients. In 30 patients, change in clinical status and CEA values occurred simultaneously. In only 2 patients an increase in CEA value occurred 2--3 months before clinical documentation of relapse.

Role of plasma carcinoembryonic antigen in evaluating patients with breast cancer treated with adjuvant chemotherapy.

Serial plasma carcinoembryonic antigen (CEA) levels were determined in 84 patients with breast cancer who were receiving postsurgical adjuvant chemoimmunotherapy or adjuvant chemotherapy. CEA values were correlated with clinical status, scintiscans, alkaline phosphatase, LDH, and SGOT. CEA values greater than 2.5 ng/ml were considered abnormal. Thirty patients had normal serial CEA values; all remain disease-free. In 54 patients one or more abnormal CEA values were recorded; nine of 54 developed overt metastatic disease. Relapses occurred in four of 38 patients in whom values up to 5 ng/ml were recorded, in three of 14 patients in whom values up to 10 ng/ml were recorded, and in two of two patients in whom values greater than 10 ng/ml were recorded.

Carcinoembryonic antigen as a marker in patients with breast cancer receiving postsurgical adjuvant chemotherapy.

Serial plasma CEA levels were determined over a period of 1-3 years in 114 patients receiving adjuvant chemotherapy for T1, 2 or 3a N+ MO breast cancer. CEA values were correlated with clinical status, scintiscans, and other biochemical parameters. CEA values greater than 2.5 ng/ml were considered abnormal. Forty-one patients had normal values throughout the adjuvant period. In 73 patients where abnormal values occurred, four different patterns were seen: (1) a statistically significant number (30/73) had initial elevations with a decreasing titer; (2) rising titer (10/73); (3) fluctuating titer of transient elevation (25/73); and (4) persistent elevation (8/73). Seventeen patients developed overt metastases, this was associated with a rising CEA in nine patients (P less than 0.002). CEA was more sensitive for predicting relapse than alkaline phosphatase or LDH. The correlation between CEA determination and the eventual development of metastatic disease is striking, and has implications for the design of future clinical trials. Results indicate that CEA could be used to identify high risk patients, to estimate efficacy of chemotherapeutic regime, and to determine optimal duration of therapy.

Ultrastructural study of poxvirus causing myxomatosis in rabbits, in São Paulo and Santa Catarina, Brazil / Estudio ultraestructural del poxvirus causal de mixomatosis en conejos, en São Paulo y Santa Catarina, Brasil

The myxomatosis is a contagious worldwide disease caused by poxvirus which infects domestic and wild rabbits. In the present study we present two distinct outbreaks of myxomatosis when raising rabbits, one for commercial purpose of production of meat and skins and, another one for the commercialization of ornamental rabbits. The observed signs were ocular, auricular, nasal, testis lesions and many times scattered throughout the body of the animals. The lesions were characterized by formation of nodules that by palpation disclosed gummy or gelatinous aspect. At the transmission electron microscopy, all the skin and crust samples were analyzed by negative staining technique. A great number of particles with morphology similar to the poxvirus, some enveloped in a brick-shaped and irregular disposition of tubules on the external membrane, measuring 300x240 nm on the average were visualized. Ultra thin sections revealed the presence of intracytoplasmic inclusion bodies surrounded by membrane containing oval particles, measuring 270 x 130 nm, containing nucleus or an internal biconcave (dumbbell-shaped) core. Immature particles (empty), surrounded by membrane were also observed. In addition, intracytoplasmic electron dense inclusion bodies containing viral particles budding of dense amorphous material and intranuclear fibrillar or "digital" inclusions showing a regular striation and arranged in groups were found in the middle of granular material. The nuclei were deformed with densely condensed chromatin forming amorphous and electron dense inclusion bodies. In the immunocytochemistry technique, the antigen-antibody reaction was strongly marked by the particles of colloidal gold, emphasizing the viral particles. The techniques used in this study were important in the diagnosis of the affected animals.

La mixomatosis es una enfermedad contagiosa de distribución mundial, causada por poxvirus que infecta conejos domésticos y salvajes. En este estudio presentamos dos distintos surtos por mixomatosis que ocurrieron en producciones de conejos, una para fines comerciales de producción de carne y pieles y otra para el comercio de conejos domésticos. Las señales observadas fueron afecciones oculares, nasales, testiculares y, a veces, también distribuida por todo el cuerpo de los animales. Estas se caracterizaban por formación de nódulos que a la palpación tenían un aspecto gelatinoso o gomoso. En la microscopía electrónica de transmisión, por la técnica de contrastación negativa, se pudo observar en todas las muestras examinadas de piel y de costras, un gran número de partículas típicas de poxvirus, con envoltura y forma de ladrillo, mostrando disposición irregular de los túbulos sobre la membrana externa, midiendo 300 x 240 nm en el promedio. Cortes ultrafinos de fragmentos de piel y de costras revelaron la presencia de cuerpos de inclusión intracitoplasmáticas, envueltos por membrana y conteniendo partículas ovales, midiendo 270 x 130 nm, conteniendo núcleo o centro interno bicóncavo (forma de mancuernas). Partículas inmaduras (vacías) envueltas por membrana fueron observadas. También fueron analizados cuerpos de inclusión intracitoplasmáticos, electrodensos, conteniendo partículas virales brotando del material denso y amorfo. Fueron observadas inclusiones intranucleares fibrilares o "digitales" mostrando una estriación periódica y disposición en grupos en medio del material granular. Los núcleos estaban deformados con cromatina densamente condensada formando cuerpos de inclusiones electrodensas y amorfas. En la técnica de imunocitoquímica la reacción antígeno-anticuerpo fue intensamente marcada por las partículas de oro coloidal realzando fuertemente las partículas virales.

Epidermodysplasia verruciformis and malignant thymoma.

We describe a patient with a long history of skin lesions clinically and histologically consistent with epidermodysplasia verruciformis (EV) who developed malignant thymoma. HPV-9DNA was found in the macular lesion and HPV-4DNA and HPV-9DNA in the coexistent common warts. Thrombocytopenia and hypogammaglobulinaemia preceded the diagnosis of thymoma. Our patient seems to represent an example of an EV-like syndrome in immunodeficiency.

Mitoxantrone in the treatment of acute leukemia.

Forty-six patients with acute leukemia were treated with mitoxantrone as a single agent. Twenty-nine patients had relapsed and/or refractory acute leukemia. Seventeen patients with acute non-lymphatic leukemia had received no prior treatment. Twelve mg/m2 of mitoxantrone was given intravenous on five consecutive days. Treatment related side effects included bone marrow suppression, mucositis, alopecia, nausea, vomiting and infection. Cardiotoxicity was documented in 7 patients. This study reconfirms that mitoxantrone is an active agent in acute leukemia with complete response documented in 10 of 29 patients with relapsed and/or refractory acute leukemia (34% response rate, 95% confidence limits 18-53%) and complete response documented in 11 of 17 patients (65% response rate, 95% confidence limits 38-87%) with previously untreated acute nonlymphatic leukemia.

Normalization of skin appearance in a patient with scleromyxoedema after intensive chemotherapy for Hodgkin's disease.

A patient with scleromyxoedema was treated for 6 years with cytostatic drugs. During this time the skin lesions followed a fluctuating but progressive course. After 6 years she developed Hodgkin's lymphoma of the mixed cellularity type. Intensive cytostatic treatment given for Hodgkin's disease resulted in virtually complete disappearance of the scleromyxoedema lesions. The development of Hodgkin's disease is considered fortuitous and not due to the previous cytostatic drugs.

Aspergillus brain abscess. A case report.

The history and findings in a patient with erythroleukaemia who developed a fungal brain abscess during the agranulocytic phase of induction treatment is reported. The radiological features of fungal infection are reported, with emphasis on the importance of clinical judgement in making the diagnosis. The autopsy findings further illustrate the increasing importance of this previously very rare condition.