Morphocytochemical, immunohistochemical and ultrastructural characterization of the head kidney of fat snook Centropomus parallelus.

This study characterized the structure and the morphocytochemical, immunohistochemical and ultrastructural aspects of the head kidney (HK) of the fat snook Centropomus parallelus. The HK is enclosed by a thin capsule of connective tissue, from which fine trabeculae originate and branch into the interior of organ. In the parenchyma, there are aggregates of lymphoid cells containing populations of lymphocytes T immunopositive for CDRO45, in a nodular arrangement, around blood vessels and melano-macrophage centres. Among the cells that constituted these aggregates and surrounded them, were macrophages and monocytes, and their precursors, with strong immunopositivity for CD68, along with cells of the granulocytic lineage in various phases of maturation positive for lysozyme and PAS. Macrophages and chromaffin and interrenal cells are also present. Ultrastructurally, the HK comprises a reticulum-endothelial stroma consisting of endothelial cells, reticulocytes of the fibroblast type and macrophage type and a parenchyma with increased cellularity, principally blood cells of the erythrocytic, granulocytic, lymphocytic, monocytic and thrombocytic series.

Early nuclear alterations and immunohistochemical expression of Ki-67, Erb-B2, vascular endothelial growth factor (VEGF), transforming growth factor (TGF-beta1) and integrine-linked kinase (ILK) two days after tamoxifen in breast carcinoma.

The purpose of the present study was to evaluate breast carcinoma samples before and two days after treatment with tamoxifen in order to analyse early histopathological alterations--particularlynuclear alterations-- as well as immunohistochemical expression of Ki-67, Erb-B2, VEGF, TGF-beta1 and ILK proteins. Twenty one cases of invasive ductal and lobular breast carcinoma were studied. Patients were submitted to biopsy of the lesion and, after confirmation of the diagnosis, they received 20 mg of tamoxifen a day, beginning two days before surgery. The samples obtained during biopsy and after surgery were stained with HE for histopathological diagnosis. Estrogen receptor was positive in 18 cases and negative in 3. The immunohistochemical method was applied for the detection of Ki-67, Erb-B2, protein, vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-beta1) and integrin linked kinase (ILK). Two days after tamoxifen treatment, the following results were observed: 1) decrease in the cell volume, chomatine condensation, nucleoli less evident and clearly defined nuclear limits; 2) significant reduction in the expression of Erb-B2 protein and significant increase in the expression of TGF-beta1 protein; 3) expression of others proteins (Ki-67, VEGF and ILK) was not altered during the indicated time frame. Our results suggest that analyzing nuclear alterations and expression of Erb-B2 and TGF-beta1 proteins would be useful to assess the initial response to tamoxifen.

Immunosuppression in undernourished rats: the effect of glutamine supplementation.

OBJECTIVE: The aim of our study is to determine the effect of a 30-day-period caloric restriction (CR) upon the immune response of rats and the influence of glutamine upon mononuclear cells proliferation and cytokine production. METHODS: Male albino Wistar rats were submitted to CR receiving an amount of food equivalent to 50% of the mean amount consumed by the control animals. We measured the incorporation of [2-14C]-thymidine by lymphocytes obtained from the spleen and mesenteric lymph nodes, plasma glucose and glutamine concentration, as well as cytokine production by cultivated cells, in the presence of glutamine. RESULTS: Rats submitted to CR presented reduced body weight (49%) and decreased splenic leukocyte number. CR led to a reduction in the proliferative response of lymphocyte. Spleenocytes from CR animals produced less gamma-interferon and interleukins 1, 4 and 10 in 48 h culture than did those from control rats. The same pattern is observed in cells obtained from the mesenteric lymph nodes. The addition of glutamine 2mM to the culture medium restored spleen and mesenteric lymph node cells' proliferative response and the production of interleukin 2 by cells obtained from the spleen and from the mesenteric lymph nodes. CONCLUSIONS: The present data reinforce that undernutrition decreases in vitro immune cell function and indicates that, in such circumstances, glutamine supplementation could reverse some of the changes observed in the functionality of cultured immune cells. The presence of the amino acid at physiological concentration, however, reinforces the diversion of the immune response towards a Th(1)-like response.

Morphological and ultrastructural study of the thrombocytes and leukocyte granulocytes of Salminus maxillosus (Characiformes, Characidae).

The thrombocytes and leukocytes of 'dourado' fish Salminus maxilosus peripheral blood were studied by means of light microscopy and transmission electron microscopy techniques. In respect to the morphological and ultrastructural aspects of thrombocytes and granulocytes, the cellular elements that appear with high frequency in the light microscopy preparations stained by the Rosenfeld method, three different types of granulocytes could be differentiated at the electron microscope, based on the ultrastructural aspect of the cytoplasmic elements: type I granulocyte that possibly corresponds to the type I neutrophil described under light microscopy; type II granulocyte is possibly the cell identified as type II neutrophil by the morphological analysis; type III granulocyte is equivalent to the eosinophils described in the samples stained by Rosenfeld methods.

Aspectos morfológicos e citoquímicos dos glóbulos sangüíneos de Caiman crocodilus yacare (Daudin, 1802) (Reptilia, Crocodilia) / Morphological and cytochemical observations of blood cells of Caiman crocodilus yacare (Daudin, 1802) (Reptilia, Crocodilia)

Foram utilizados 10 jacarés Caiman crocodilus yacare, colhendo-se 5 ml do sangue periférico de cada animal. A análise morfológica foi realizada após coloraçäo por Leishman. Para estudo citoquímico, empregaram-se os métodos do PAS, do Sudan black B, da o-toluidina e do azul de bromofenol. Foram identificados 7 tipos celulares: eritrócitos, trombócitos, heterófilos, eosinófilos, basófilos, linfócitos e monócitos azurófilos. Os eritrócitos nucleados apresentam formato elíptico com atividade mitótica e positividade citoplasmática para o azul de bromofenol. Os trombócitos säo de formato elíptico, com citoplasma abundante nos pólos, contendo grânulos de glicogênio e núcleo com sulcos profundos. Heterófilos, grosseiramente esféricos, mostram núcleo esférico excêntrico e citoplasma repleto de grânulos corados em salmäo, de formato fusiforme, em baqueta, oval ou esférico. A citoquímica nestas células revelou a presença de glicogênio, grânulos citoplasmáticos positivos para azul de bromofenol e parcialmente sudanófilos e positivos para mieloperoxidase. Eosinófilos mostram-se esféricos com núcleo lenticular excêntrico e citoplasma com grânulos esféricos ou ovais róseos positivos para Sudan e mieloperoxidase, porém fracamente para o azul de bromofenol. Basófilos apresentam formato esférico, de tamanho menor em relaçäo aos demais granulócitos, núcleo esférico central e citoplasma com poucos grânulos fortemente basófilos. Linfócitos mostram-se polimórficos com núcleo de formato irregular, citoplasma escasso com projeçöes e grânulos azurófilos. Monócito azurófilo, de formato esférico, núcleo excêntrico e citoplasma basófilo contendo grânulos azurófilos abundantes

Identification and characterization of a sialidase released by the salivary gland of the hematophagous insect Triatoma infestans.

Sialidases (EC 3.2.1.18) are commonly found in viruses, bacteria, fungi, protozoa, and vertebrates, but not in invertebrates. We have previously reported the presence of a new sialidase activity in the gut of exclusively hematophagous insects of the Triatoma genus, which transmit Chagas' disease (Amino, R., Acosta, A., Morita, O. M., Chioccola, V. L. P., and Schenkman, S. (1995) Glycobiology 5, 625-631). Here we show that this sialidase is present in the salivary gland of Triatoma infestans, and it is released with the saliva during the insect bite. The sialidase was purified to homogeneity (>5000 times) to a specific activity of more than 20 units/mg. It elutes from a gel filtration column with a volume corresponding to the size of 33 kDa, and it migrates as a single 26-kDa band in SDS-polyacrylamide gel electrophoresis, which is unusually smaller when compared with other known sialidases. T. infestans sialidase hydrolyzes preferentially alpha2-->3-linked sialic acids at pH 4-8, with maximal activity between pH 5.5 and 6.5, which is compatible with the optimal pH of secreted sialidases. The sialidase is competitively inhibited by 2-deoxy-2, 3-dehydro-N-acetyl-neuraminic acid (Ki = 0.075 mM) and differently from many sialidases, with exception of Salmonella typhimurium sialidase, it is inhibited competitively by HEPES (Ki = 15 mM). The fact that T. infestans sialidase is released with the saliva and can hydrolyze sialyl-LewisX blood groups, which are the ligands for selectins, suggests that it might have a role in the blood feeding.

Estudos hematológicos em Oreochromis niloticus (Linnaeus, 1758) (Cichlidae, Teleostei) - Parte I / Hematological studies in Oreochromis niloticus (Linnaeus, 1758) (Cichlidae, Teleostei) - Part I

Avaliou-se o hemograma e a morfologia celular em amostras de sangue periférico obtidas de 10 peixes da espécie Oreochromis niloticus. Parte das mesmas foi utilizada para o estudo da hematimetria, isto é, contagem de eritrócitos (x = 2,35 x 10 elevado à sexta potência/mm ao cubo mais ou menos 0,12), taxa de hemoglobina (x = 7,04g/dL mais ou menos 0,38), hematócrito (x = 27,85 por cento mais ou menos 1,62), HCM (x = 30,31pg mais ou menos 1,64), VCM (x = 118,6u ao cubo mais ou menos 3,53), CHCM (x = 25,78 por cento mais ou menos 1,72), contagem global de leucócitos (x = 16,08 x 10 ao cubo/mm ao cubo mais ou menos 1,28), contagem de trombócitos (x = 61,69 x 10 ao cubo/mm ao cubo mais ou menos 5,96), sendo a avaliaçäo da morfologia das células do sangue realizada através do estudo de extensöes sanguíneas submetidas à coloraçäo de Wright. Os eritrócitos e trombócitos nucleados apresentaram-se elípticos; neutrófilos e eosinófilos esféricos de tamanho variável, com o núcleo também esférico, apresentaram citoplasmas abundantes, ligeiramente basófilo e altamente acidófilo, respectivamente. Näo foram vistas as formas polimorfonucleares dessas células. Os basófilos apresentaram núcleo esférico e granulaçöes metacromáticas. Os monócitos e os linfócitos assemelharam-se morfologicamente aos dos mamíferos

Açäo dos raios X corpo total sobre os neutrófilos do sangue periférico em primata Cebus apella (macaco prego) / Effects of whole-body X-radiation on the neutrophils of the peripheral blood of the primate Cebus apella (weeping capuchin)

Amostras de sangue periférico foram obtidas de cinco primatas Cebus apella (macaco prego), antes e após exposiçäo aos raios X corpo inteiro, na dose única de 25,8 mC/kg (100R). Parte das amostras foi utilizada para a contagem global de cariosquizes e o restante para a realizaçäo de extensöes sanguíneas e submetidas à coloraçäo de Wright e aos seguintes métodos citoquímicos: ácido periódico Schiff (PAS) para a identificaçäo de glicogênio, Sudan black B para a detecçäo de fosfolipídios e o-toluidina-água oxigenada para a demonstraçäo da mieloperoxidase. Com o método de Wright foram observadas aos 1, 3 e 6 dias após exposiçäo à radiaçäo, alteraçöes nucleares e citoplasmáticas. Após aplicaçäo dos métodos citoquímicos acima referidos, foram observados, de modo geral, nos mesmos períodos de tempo após irradiaçäo: alteraçäo no padräo de distribuiçäo dos grânulos de glicogênio, grânulos mieloperoxidase positivos e sudanófilos de neutrófilos. No que diz respeito a cariosquizes, constatou-se um aumento dos mesmos após 1 dia, persistindo até o 3§ dia após irradiaçäo. Noventa dias após, näo foi observada nenhuma alteraçäo acima referida

Effect of fatty acid-rich diets on thymocyte proliferation and thymus involution during growing.

The effect of poly-unsaturated and saturated fatty acid-rich diet on the proliferation response of T and B lymphocytes in vitro was investigated. Also, a mitotic index of thymocytes and of thymus epithelial reticular-like cells and thymic involution coefficient were examined in vivo. w-6 PUFA-rich diet enhanced the rate of T (eightfold) and B (fourfold) lymphocyte proliferation, whereas saturated fatty acids did not show an important effect. However, both fat-rich diets increased the mitotic index of thymocytes and thymus involution index in vivo. These findings suggest that fat-rich diets may accelerate thymus involution during growing, possibly aggravating the impairment of the immune function which usually occurs under this condition.

Effects of whole-body x-irradiation on the peripheral blood of primate Cebus apella

The effects of ionizing radiation on the peripheral blood elements of primate Cebus apella were examined after a single 25.8mC/kg (100 R) dose of whole-body X-ray. We determined the number of white blood cells,neutrophils, osophils, basophils,lymphocytes,monocytes anderythrocytes up to 90 days after exposure of 5 animals. Hemoglobin content and microhematocrit were also determined. Under these experimental conditions, the total number of leukocytes was reduced from 9440/mm***3 to 5660/mm***3 on day6, mainly because of a decrease inthe lymphocyte population from 5843.2/mm***3 to 2349.0/mm***3 on day 6. No significant differences were observed in the values of erythrocytes, neutrophils,eosinophils,basophils,monocytes,hemoglobin content and microhematocrit. All hematological parameters studied returned tonormal values by day 90 after a single irradiation dose of 100R

Topochemistry of blood cells of the Gallus domesticus (Aves, Galliforme).

Blood smears of both male and female chicken Gallus domesticus were analysed by using the following topochemical methods: a) Periodic acid-Schiff (PAS) for glycogen. b) Mercury-bromophenol blue for protein. c) O-Toluidine for myeloperoxidase. d) Sudan black B for lipid. The PAS reaction revealed glycogen in the cytoplasm of all thrombocytes and in a few heterophils. The presence of proteins was evidenced in all types of cells. However variation in the intensity of staining of protein granules was observed in the fusiform structures of the heterophils. A negative reaction for myeloperoxidase was found in all cells. Although some evidence of myeloperoxidase activity was show in the polymorphonuclears it was not enough to ascertain a positive reaction. Lipids were detected in the cytoplasm of few heterophils, eosinophils and monocytes.

Topochemistry of blood cells of the Gallus domesticus (Aves, Galliforme)

Extensöes de sangue periférico de aves (macho e fêmea), foram submetidas a um estudo topoquímico que permitiu as seguintes conclusöes: com o PAS, a presença de glicogênio nos trombócitos e alguns heterófilos; com o azul de bromofenol (proteína) positividade em todos os elementos celulares, com intensa reatividade da regiäo equatorial dos grânulos fusiformes de heterófilos; com o método da orto-toluidina-água oxigenada (mieloperoxidase), reatividade discutível nos polimorfonucleares e negativa nos demais tipos de células e com Sudan black (lipídios) uma positividade em alguns heterófilos, eosinófilos e monócitos

Effects of whole-body X-irradiation on the peripheral blood of primate Cebus apella.

The effects of ionizing radiation on the peripheral blood elements of primate Cebus apella were examined after a single 25.8 mC/kg (100 R) dose of whole-body X-ray. We determined the number of white blood cells, neutrophils, eosinophils, basophils, lymphocytes, monocytes and erythrocytes up to 90 days after exposure of 5 animals. Hemoglobin content and microhematocrit were also determined. Under these experimental conditions, the total number of leukocytes was reduced from 9440/mm3 to 5660/mm3 on day 6, mainly because of a decrease in the lymphocyte population from 5843.2/mm3 to 2349.0/mm3 on day 6. No significant differences were observed in the values of erythrocytes, neutrophils, eosinophils, basophils, monocytes, hemoglobin content and microhematocrit. All hematological parameters studied returned to normal values by day 90 after a single irradiation dose of 100 R.

Histochemical study of labial salivary glands in Sjögren's syndrome.

Human labial salivary gland biopsies of patients presenting connective tissue diseases associated with Sjögren's syndrome were submitted to a polysaccharide histochemistry study. The normal acinar secretion is an association of neutral polysaccharides with a sulphosialomucin. In Sjögren's syndrome, there is a great reduction in the secretory activity of the acinar cells, but no qualitative change was observed. The pathogenesis of this decreased production and its importance regarding the clinical manifestations of Sjögren's syndrome are discussed.