Comparação entre dois protocolos para obtenção de plasma rico em plaquetas, em cães / Comparison between two protocols to obtain platelet-rich plasma in dogs

Avaliaram-se dois protocolos para a produção de plasma rico em plaquetas (PRP) com o sangue de 20 cães adultos. Foram coletados três frascos de sangue em que um deles foi usado para produção do PRP por meio do protocolo A - centrifugação única a 1200rpm/10min -, o outro para fabricação do PRP pelo protocolo B - primeira centrifugação a 1200rpm/10min e a segunda centrifugação a 1600rpm/10min - e o terceiro para realização da contagem plaquetária no sangue total, que serviu de parâmetro para os valores alcançados no PRP. O protocolo no qual foi possível alcançar maior concentração plaquetária foi testado em outros 20 cães para avaliar sua reprodutibilidade. Constatou-se que o protocolo B resultou em maior plaquetometria em 100 por cento das amostras e concluiu-se ser ele eficiente para a produção do PRP em cães.

The objective of this paper was to analyze two protocols for the production of platelet rich plasma (PRP) in dogs. Peripheral blood of 20 adult dogs was collected into three tubes. The first was processed through protocol A - single centrifugation at 1200rpm for 10min - , the second was submitted to protocol B - a first centrifugation at 1200rpm for 10min and a second centrifugation at 1600rpm for 10min - and the third was used to perform platelet count in whole blood, which served as a parameter for values obtained in PRP. The protocol in which it was possible to achieve a higher platelet count was tested in other 20 dogs to evaluate its reproducibility. Protocol B resulted in a superior platelet count in 100 percent of the samples, concluding that the referred protocol is effective for PRP production in dogs.

Study of irradiated bothropstoxin-1 with60Co gamma rays: immune system behavior

Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules. Very promising results were obtained when crude animal venoms, as well as isolated toxins, were treated with 60Co gamma rays, yielding toxoids with good immunogenicity. The achievement of modified antigens with lower toxicity and preserved or improved immunogenicity can be very useful. Ionizing radiation has already been proven to be a powerful tool to attenuate snake venom toxicity without affecting, and even increasing, their immunogenic properties. However, little is known about the modifications that irradiated molecules undergo and even less about the immunological response that such antigens elicit. In the present work, we investigated the immunological behavior of bothropstoxin-1, a K49 phospholipase, before and after irradiation. Structural modifications of the toxin were analyzed by SDS-PAGE. Isogenic mice were immunized with either the native or the irradiated toxin. The circulating antibodies were isotyped and titrated by ELISA. According to our data, irradiation promoted structural modifications in the toxin characterized by higher molecular weight forms of proteins (aggregates and oligomers). The results also indicated that irradiated toxins were immunogenic and antibodies elicited by them were able to recognize the native toxin in ELISA. These findings suggest that irradiation of toxic proteins can promote significant modifications in their structures; however they still retain many of the original antigenic and immunological properties of native proteins. Also, our data indicate that irradiated proteins induce higher titers of IgG2a and IgG2b, suggesting that Th1 cells are predominantly involved in the immune response.

Evidence for the participation of nitric oxide in pemphigus

Pemphigus is an inflammatory autoimmune disorder of the skin. Nitric oxide (NO) is an inflammatory mediator linked to a variety of physiological and pathophysiological phenomena that include skin tumors, psoriasis, urticaria, and atopic dermatitis. Inflammatory cells present in pemphigus lesions are important sources of NO production. We investigated whether NO is involved in pemphigus. A prospective cohort study was conducted at the Dermatology Service of the Hospital Universitário Walter Cantídio of the Federal University of Ceará. All patients seen at the outpatient clinic between August 2000 and July 2001, with a clinically and histologically confirmed diagnosis of pemphigus were included. The median age was 42.5 years (range: 12-69 years) with a male to female ratio of 3:2. Total serum nitrite levels, used as a marker for NO production, were determined by the Griess reaction. Skin biopsies from pemphigus and breast surgery (control) patients were used for the detection of the inducible NO synthase (iNOS) by immunohistochemistry. Twenty-two (22) patients with pemphigus and eight (8) controls who did not differ in demographic characteristics were included. Total serum nitrite levels were significantly higher (>7 æmol/L) in pemphigus patients compared to controls (<6 æmol/L), regardless of the severity of the clinical activity of pemphigus (P < 0.0001). All pemphigus biopsies presented increased immunostaining for iNOS that was not detected in normal skin samples. These data are the first to demonstrate that pemphigus patients display increased serum NO levels that are associated with increased iNOS expression in the affected skin.

Hipertensao arterial e estado de alcoolismo. / Arterial hypertension and alcoholism

Foram estudados 2.980 casos de alcoolismo no PSM de Juiz de Fora (MG); todos os casos apresentando sinais de ingesta recente de bebida alcoolica, motivo pelo qual procuraram assistencia medica. A hipertensao arterial (HA) foi encontrada em 68 individuos (2,28%), perfazendo 56 homens e 12 mulheres; 19 brancos, 40 pretos e 9 mulatos. A maioria da populacao estudada era jovem (15-35 anos), o mais jovem hipertenso com 15 anos e o mais velho com 65 anos. A HA foi mais frequente nos individuos pretos entre 26 e 45 anos (p< 0,05). 41,17% apresentaram-se hipertensos com alcoolismo tipo agudo e 58,83% hipertensos com alcoolismo tipo cronico. Conclui-se que a HA na populacao alcoolatra estudada coincide com a maior frequencia da HA essencial que incide sobre a raca masculina jovem