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Melanomacrophages are phagocytes that synthesize melanin. They are found in the liver and spleen of ectothermic vertebrates, and in the kidney of fish. In agnathan and elasmobranch fish, melanomacrophages are seen as isolated cells, and forming clusters in all the other vertebrates. The natural phagocytic activity of melanomacrophages is poorly characterized, as most of the research works have focused on induced phagocytic activity only. Furthermore, little is known about amphibian melanomacrophages, mainly about those in caecilians – wormlike amphibians in the order of Gymnophiona, which is the least known group of terrestrial vertebrates. The present research work aimed at the structure and function of hepatic melanomacrophages of Siphonops annulatus, a species largely found in South America. We identified the role of these cells in the control of circulating basophils (pro‐melanogenic cells), in the turnover of liver collagen stroma and in the hemocatheresis, interrelated physiological mechanisms.

RESUMO: Os animais utilizados neste trabalho foram fornecidos pelo Departamento de Parque de Área Verde (SP). As amostras de sangue foram colhidas entre agosto de 1999 a novembro de 2001 e determinados os seguintes parâmetros hematológicos: contagem total de eritrócitos (Er), hematócrito (Ht), taxa de hemoglobina (Hb), contagem total e diferencial de leucócitos, índices hematimétricos absolutos VCM e CHCM. Foi realizada também a descrição morfológica das células sanguíneas em extensões sanguíneas após aplicação do método de Leishman. Os valores referentes à série eritrocítica apresentaram as seguintes médias: Er= 2,62 ± 0.48 x 106/µL; Ht=43,16 ± 11,19%; Hb= 14,59 ± 3,38g/dl; VCM=166,33 fL; CHCM= 33,84 ± 2,55g/dl e àqueles referentes a contagem de leucócitos apresentaram média: Lc= 3,95 ± 2,06 x 103/µL; Ht=45,33 ± 9,48%; Eo=17,66 ± 10,40%; B=7,3 ± 3,05%; L= 27,33 ± 11,53% e M= 2 ± 2,30%. Foram observados eritrócitos elípticos, com núcleo heterocromático alongado e citoplasma acidófilo; trombócitos elípticos com núcleo basófilo, cromatina condensada e citoplasma hialino; heterófilos esféricos, com núcleo lobulado e presença de grânulos acidófilos fusiformes; eosinófilos esféricos com núcleo lobulado apresentando grânulos eosinófilos esféricos; basófilos com núcleo volumoso e grânulos basófilos; linfócitos de aspecto irregular, com núcleo esférico e grânulos azurófilos; monócitos esféricos, com núcleo reniforme, cromatina frouxa, nucléolo visível e grânulos azurófilos finos.Palavras-chave: Gavião Carijó; Buteo magnirostris; Hematologia. ABSTRACT: The animals used in this study were provided by the Green Area Park Department (SP). Blood samples were collected from August 1999 to November 2001 and determined the following hematological parameters: total count of erythrocytes (Er), hematocrit (Ht), hemoglobin (Hb), total count and differential leukocyte count, erythrocyte indices absolute MCV and MCHC. It also performed the morphological description of the blood cells in blood smears after application of Leishman method. The figures for the erythrocyte series presented the following averages: Er = 2.62 ± 0:48 x 106 / µL; Ht = 43.16 ± 11.19%; Hb = 14.59 ± 3,38g / dl; VCM = 166.33 fL; MCHC = 33.84 ± 2.55g / dl and those related to leukocyte count showed average: Lc = 3.95 ± 2.06 x 103 / µL; Ht = 45.33 ± 9.48%; And = 17.66 ± 10.40%; B = 7.3 ± 3.05%; L = 27.33 ± 11.53% and M = 2 ± 2.30%. erythrocytes were observed elliptical, with elongated heterochromatic nucleus and cytoplasm acidophilus; thrombocytes Elliptical with basophil nucleus, condensed chromatin and hyaline cytoplasm; Spherical heterophile with lobed nucleus and presence of acidophilic spindle granules; eosinophil-lobed spherical core having eosinophils spherical granules; basophils with large nuclei and basophil granules; irregular aspect of lymphocytes, with spherical core and azurophilic granules; Spherical monocyte, a kidney-shaped core, loose chromatin and nucleoli visible fine azurophilic granules.Keywords: Sparrowhawk Carijó; Buteo magnirostris; Hematology.

The objective of this work was to determine the hematological parameters and the phagocytic capacity of peritoneal macrophages of fat snook related to sex, stage of gonadal maturation and seasonal cycle. Blood was collected from 135 animals (78 females and 57 males) and used for determinations of: erythrocyte number, hematocrit, hemoglobin, erythrocyte indices mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), total and differential leukocyte counts, and thrombocyte count. The phagocytic capacity and phagocytic index were determined after Saccharomyces cerevisiae inoculation in the peritoneal cavity of the animals. The hematological results according to sex showed that the erythrocyte, total leukocyte and thrombocyte counts were statistically higher in males than females, with the latter showing a higher MCV. Concerning to erythrocyte count, hematocrit and hemoglobin concentration analyzed separately by sex and stage of gonadal maturation, males were found to have significantly elevated values in the mature stage and decreased levels in the resting stage. The results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to seasonal cycle showed significant differences in both sexes, where hematocrit and hemoglobin concentration were lower in winter and higher in the other seasons, mean corpuscular volume was higher in the summer and lower in the winter and fall, total leukocytes and thrombocytes lower in the spring and higher in the fall, lymphocytes low in the winter and summer and high in the spring and phagocytic capacity and phagocytic index high in the summer and low in the winter and fall. The results showed that the hematological values in males are statistically higher than those in females, the erythrocyte values in males increase with the progression of gonadal maturation and that winter is the season of the year least favorable for hematological and phagocytic responses for survival of fat snook kept in captivity. The parameters studied could be utilized in the evaluation of the health status of this species in captivity.

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.

We studied p38 phosphorylation and its intracellular localization during p53 and Puma (a p53 upregulated modulator of apoptosis) apoptotic signaling pathway in bone marrow granulocytes in mice irradiated in vivo and the role of the radioprotector amifostine in ameliorating these responses. Sixty-four C57BL mice were randomly assigned in two non-irradiated (Ami−/rad− and Ami+/rad−) and two irradiated (Ami−/rad+ and Ami+/rad+) groups. Animals received 400 mg/kg of amifostine i.p. 30 min prior to a single whole body radiation dose of 7 Gy. The experiments were performed using immunohistochemistry for caspase-3, cleaved caspase-3, p53, p-p53 (Ser 15), Puma, p38 and p-p38 (Thr 180/Tyr 182) protein expression. In addition transmission electron microscopy was used for ultrastructural characterization of apoptosis. Data showed that: (i) amifostine significantly reduced the number of apoptotic cells, (ii) p-p53 and Puma proteins were strongly immunostained in granulocytes after irradiation (Ami−/rad+), (iii) amifostine decreased the immunostaining of the proteins (Ami+/rad+), (iv) p38 was immunolocalized in physiological conditions in the nucleus and cytoplasm of granulocytes and neither radiation nor amifostine changed the protein immunostaining or its subcellular distribution, but influenced its activation, (v) radiation-induced p38 phosphorylation and its cytoplasmic accumulation during apoptosis signaling in granulocytes after whole body high radiation dose and amifostine markedly reduced these effects.

The rudimentary characteristic of the eyes of fossorial animals raises some questions regarding its evolution and functionality. Would these eyes result from atrophy or from stagnated development? How would its visual function work? Anatomical investigations of these organs are the fundamental preamble to answer those questions, which are still little explored by the literature. in this article we have studied anatomical aspects of the eyes of three species of fossorial reptiles, within the suborder Amphisbaena (Amphisbaena alba, Amphisbaena mertensi, Leposternon infraorbitale), as well as a species within the ophidian suborder (Typhlops brongersmianus). the minuscule eyes (1-2 mm diameter) were visualized through a scale, a translucent area which corresponds to the spectacle. This spectacle is a thinner and transparent scale, covering a conjunctival sac. the retrobulbar space was filled with the harderian gland. the eyes of Typhlops presented an oval shape, whereas Amphisbaena specimens presented cup-shaped eyes. in Amphisbaenian sclera is comprised of cartilage, while the thin sclera of Typhlops consists of connective tissue and striated muscle fibers. the retina presented all the typical layers found in vertebrates, regardless the species. the characteristics involved in the fossil adaptation of these species include: reduced size of the eyeball, rudimentary cornea, absence of the anterior chamber, presence of a complex iris-ciliary body, and lens with amorphous nucleate cells. the analysis of the eye morphology of these animals suggests that there might be a specific function concerning light perception. Anat Rec, 293:351-357, 2010. (C) 2009 Wiley-Liss, Inc. Universidade Federal de São Paulo, Disciplina Anat Descrit & Topog, Dept Morfol & Genet, Escola Paulista Med, BR-04023900 São Paulo, Brazil Inst Butantan, Lab Biol Celular, São Paulo, Brazil Universidade Federal de São Paulo, Disciplina Anat Descrit & Topog, Dept Morfol & Genet, Escola Paulista Med, BR-04023900 São Paulo, Brazil Web of Science

The objective of this work was to determine hematological parameters and the phagocytic capacity of peritoneal macrophages of the fat snook related to sex, stage of gonadal maturation and seasonal cycle. The hematological results did not show significant differences between males and females. With respect to stage of gonadal maturation, only erythrocytes numbers (RBC), in females, was found to be significantly elevated in stage III (maturing) and decreased in V (spent). An analysis of the results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to sex and seasonal cycle showed statistically significant differences: a) hematocrit (Ht) and hemoglobin concentration (Hb) were elevated in the spring and low in the winter in males; b) mean corpuscular volume (MCV) was high in the spring and summer and low in the fall in both sexes; c) thrombocytes numbers were elevated in the fall and low in the other seasons in females; d) phagocytic capacity (PC) and phagocytic index (PI) were higher in the summer and lower in the fall in females. The results showed that spring and summer correspond to seasons of the year for better hematological and phagocytic responses for survival of the fat snook in its natural habitat. The parameters studied could be utilized for evaluation of the health status fat snook in its own habitat or in captivity.

The caecilians (Amphibia, Gymnophiona) constitute one of the least known groups of terrestrial vertebrates because most species live underground in quite inaccessible environments. Siphonops annulatus is an exclusively fossorial species and is the most extensively distributed caecilian in South America. Little is known of this order concerning circulating granulocytes, including their morphological and cytochemical structure and ultrastructure. This paper is part of a project covering the study of granulocytes in representative species of the order Amphibia. Blood extensions were carried out and submitted to Leishman, Toluidine Blue, Periodic acid Schiff, Sirius Red and hydrogen o-toluidine peroxide methods. Part of the samples was prepared for conventional transmission electron microscopy. Among granular leukocytes, mature and immature neutrophils and eosinophils were identified, plus basophils. The most frequent granulocyte encountered in S. annulatus peripheral blood is the neutrophil. This is a cell with a hyper-segmented nucleus and with a very clear cytoplasm when compared to the eosinophil, which presents large cytoplasmic acidophilic granules. On the other hand, the basophils present basophilic and metachromatic granules. Glycogen was detected in the cytoplasm of the neutrophils and eosinophils, while basic protein rich in amino acids was observed in the eosinophil’s granules. Myeloperoxidase activity was detected in the cytoplasm of the neutrophils and eosinophils. Neutrophils were ultrastructurally detected with three types of small granules: eosinophils with large and small spherical granules and basophils with large spherical granules with lamellate structures.

In the present work features of tick-bite lesions were evaluated in capybaras naturally infested with Amblyomma cajennense and Amblyomma dubitatum ticks. Gross appearance of tick bite site was characterized by a mild swelling and erythema. Microscopic examination revealed the cement cone, a tube-like homogenous eosinophilic mass penetrating deep into the dermis. This structure was surrounded in the dermis by a cellular infiltrate and free eosinophilic granules and was associated to edema of variable intensity. Necrosis was a common feature deep in the dermis particularly at the far end of the eosinophilic tube. Hyperplasia, cellular edema and occasionally necrosis of keratinocytes could be seen at both sides of the ruptured epidermis. Cellular infiltrate was constituted overwhelmingly by polymorphonuclear leukocytes with eosinophilic granules. In capybaras cells with such features can be either eosinophils or heterophils (pseudoeosinophils), the latter being the equivalent of neutrophils of other mammals. Ultrastructural analysis of the cellular infiltrate revealed the predominance of heterophils over eosinophils. Mononuclear cells and mast cells and, in lesser numbers, basophils were also seen at skin attachment sites. The presence of heterophils in the reaction of capybaras against Amblyomma ticks is an outstanding feature but its role in the reaction to the tick is not known. It is however speculated that capybara heterophils might be associated with a more permissive environment for tick feeding and pathogen transmission as already shown for the equivalent cell type, the neutrophil, in the reaction of the dog against the Rhipicephalus sanguineus tick.

OBJETIVO: Avaliar, no coração, por imuno-histoquímica, a localização das proteínas TGFbeta1 latente e TGFbeta1 ativa, se ocorre ativação radioinduzida da proteína TGFbeta1 latente, e a distribuição das fibras colágenas em diversos períodos de tempo após irradiação. MATERIAIS E MÉTODOS: Trinta e dois camundongos isogênicos (C57BL) foram divididos em dois grupos: GI (não irradiado), com 12 animais, e GII (irradiado), com 20 animais. Os animais do GII receberam radiação gama (telecobaltoterapia, 60Co, com rendimento de 0,97 Gy/min., dose única de 7 Gy em corpo inteiro). Os camundongos dos grupos I e II foram sacrificados por estiramento cervical nos períodos de 1, 14, 30 e 90 dias após irradiação. RESULTADOS: Os corações irradiados apresentaram: 1) alterações nucleares e diminuição das estriações das células musculares cardíacas; 2) aumento significante da deposição de fibras colágenas aos 90 dias depois da irradiação; 3) ativação da proteína TGFbeta1 latente em cardiomiócitos e células do conjuntivo depois da irradiação. CONCLUSÃO: Nossos resultados mostram a importância da proteína TGFbeta1 no processo de fibrose cardíaca radioinduzida e sugerem que células do parênquima (cardiomiócitos) e do conjuntivo podem participar deste mecanismo atuando como fontes da proteína TGFbeta1 ativa. OBJECTIVE: To assess the latent and active TGFbeta1 localization in the heart, to evaluate whether or not radiation induces latent TGFbeta1 activation, and to study the distribution of collagen fibers in the irradiated heart. MATERIALS AND METHODS: Thirty-two C57BL mice were randomly assigned in two groups: GI (non irradiated animals) and GII (irradiated animals). The mice from GII received a single whole-body radiation dose of 7Gy, using a 60Co source at a dose rate of 0.97 Gy/min. The animals were sacrificed by cervical dislocation at 1, 14, 30 and 90 days after irradiation. RESULTS: The irradiated hearts showed: 1) nuclear changes and muscle cells with decreased striations; 2) significant increase in the collagen deposition 90 days after irradiation; 3) latent TGFbeta1 activation in the cardiomyocytes and connective tissue cells after irradiation. CONCLUSION: Our results show the importance of TGFbeta1 protein in the process of radiation-induced heart fibrosis and suggest that cardiomyocytes and connective cells may play a role in this mechanism acting as cellular sources of active TGFbeta1.

OBJETIVO: Avaliar, no coração, por imuno-histoquímica, a localização das proteínas TGFbeta1 latente e TGFbeta1 ativa, se ocorre ativação radioinduzida da proteína TGFbeta1 latente, e a distribuição das fibras colágenas em diversos períodos de tempo após irradiação. MATERIAIS E MÉTODOS: Trinta e dois camundongos isogênicos (C57BL) foram divididos em dois grupos: GI (não irradiado), com 12 animais, e GII (irradiado), com 20 animais. Os animais do GII receberam radiação gama (telecobaltoterapia, 60Co, com rendimento de 0,97 Gy/min., dose única de 7 Gy em corpo inteiro). Os camundongos dos grupos I e II foram sacrificados por estiramento cervical nos períodos de 1, 14, 30 e 90 dias após irradiação. RESULTADOS: Os corações irradiados apresentaram: 1) alterações nucleares e diminuição das estriações das células musculares cardíacas; 2) aumento significante da deposição de fibras colágenas aos 90 dias depois da irradiação; 3) ativação da proteína TGFbeta1 latente em cardiomiócitos e células do conjuntivo depois da irradiação. CONCLUSÃO: Nossos resultados mostram a importância da proteína TGFbeta1 no processo de fibrose cardíaca radioinduzida e sugerem que células do parênquima (cardiomiócitos) e do conjuntivo podem participar deste mecanismo atuando como fontes da proteína TGFbeta1 ativa.OBJECTIVE: To assess the latent and active TGFbeta1 localization in the heart, to evaluate whether or not radiation induces latent TGFbeta1 activation, and to study the distribution of collagen fibers in the irradiated heart. MATERIALS AND METHODS: Thirty-two C57BL mice were randomly assigned in two groups: GI (non irradiated animals) and GII (irradiated animals). The mice from GII received a single whole-body radiation dose of 7Gy, using a 60Co source at a dose rate of 0.97 Gy/min. The animals were sacrificed by cervical dislocation at 1, 14, 30 and 90 days after irradiation. RESULTS: The irradiated hearts showed: 1) nuclear changes and muscle cells with decreased striations; 2) significant increase in the collagen deposition 90 days after irradiation; 3) latent TGFbeta1 activation in the cardiomyocytes and connective tissue cells after irradiation. CONCLUSION: Our results show the importance of TGFbeta1 protein in the process of radiation-induced heart fibrosis and suggest that cardiomyocytes and connective cells may play a role in this mechanism acting as cellular sources of active TGFbeta1.

Amostras de sangue foram colhidas de 37 capivaras parasitadas por carrapatos e de 21 capivaras livres de infestação para a identificação dos diferentes tipos celulares circulantes e para realização de hemograma. Após análise morfológica, as células sangüíneas foram identificadas como heterófilos, eosinófilos, basófilos, linfócitos, monócitos e plaquetas. A reação de PAS demonstrou glicogênio no citoplasma dos heterófilos e plaquetas. Reação positiva para mieloperoxidase e lipídeos foi demonstrada apenas nos grânulos citoplasmáticos esféricos dos heterófilos e em todos os grânulos citoplasmáticos dos eosinófilos. O método do azul de bromofenol revelou reação fortemente positiva nos grânulos de heterófilos, eosinófilos, basófilos e nas hemácias. Reação positiva para fosfatase alcalina foi observada nos heterófilos e eosinófilos. Os valores hematimétricos que mostraram diferenças estatisticamente significantes (p< 0,05), quando comparados aos animais parasitados com aqueles livres de infestação, foram: contagem total de hemácias (3,13±0.31 x 106/µL e 2,81±0,62 x 106/µL), hematócrito (48.14±4.75% e 41,22±6.21%), e, na contagem diferencial, os heterófilos (39,38±11,02% e 16,14±7,13%) e, eosinófilos (8,57±3,78% and 21,92±10,24%). Os carrapatos foram identificados como Amblyomma cajenense and Amblyomma cooperi. As capivaras altamente infestadas por esses carrapatos apresentaram anemia e uma eosinofilia marcante.Blood samples were collected from thirty seven adult parasited capybaras and twenty one free of parasitism aiming the characterization of different circulating cell types and complete hemogram. After a morphological analysis, the blood cells were identified as heterophils, eosinophils, basophils, lymphocytes, monocytes and platelets. The PAS reaction revealed glycogen in heterophils and platelets cytoplasm. A positive reaction was found only in heterophils round granules and in all cytoplasmic granules of eosinophils for the demonstration of myeloperoxidase activity and lipids. The bromofenol blue method revealed strong positivity in heterophils, eosinophils, basophils granules and red blood cells. A positive reaction for alkaline phosphatase was observed in eosinophils and heterophils. The blood parameters studied showed statistic differences (p< 0,05) between non- parasited capybaras and parasited capybaras in the red blood cell count (3,13±0.31 x 106/µL and 2,81±0,62 x 106/µL) , haematocrit (48.14±4.75% and 41,22±6.21%) and heterophils (39,38±11,02% and 16,14±7,13%) /eosinophils (8,57±3,78% and 21,92±10,24%) in differential count. In this case the ticks involved in the parasitism of these capybaras were identified as Amblyomma cajenense and Amblyomma cooperi. These data showed that the highly infested capybaras presented some anemia and a remarkable eosinophilia.

OBJETIVO: Investigar no pulmão, por imunohistoquímica, a localização das proteínas TGFbeta1 latente e TGFbeta1 ativa, se ocorre ativação radioinduzida da proteína TGFbeta1 latente e a distribuição das fibras colágenas em diversos períodos de tempo após irradiação. MÉTODOS: 32 camundongos isogênicos (C57BL) foram divididos em dois grupos: GI (não irradiado) com 12 animais e GII (irradiado) com 20 animais. Os animais do GII receberam radiação gama (telecobaltoterapia, 60Co, com rendimento de 0,97Gy/min, dose única de 7Gy em corpo inteiro). Os camundongos dos grupos I e II foram sacrificados por estiramento cervical nos períodos de 1, 14, 30 e 90 dias após irradiação. RESULTADOS: Os pulmões irradiados apresentaram: 1) congestão vascular e espessamento dos septos alveolares aos 30 dias e mais intensamente aos 90 dias depois da irradiação; 2) aumento significante da deposição de colágeno em todos os períodos de tempo após irradiação; 3) fraca ativação da proteína TGFbeta1 latente em um dia e intensa aos 14 dias depois da irradiação em brônquios e alvéolos. Nossos resultados sugerem que células dos brônquios e alvéolos podem participar do complexo mecanismo de fibrose pulmonar radioinduzida atuando como fontes da proteína TGFbeta1 ativa.PURPOSE: assess the latent and active TGFb1 localization in the lung, whether or not radiation induces latent TGFbeta1 activation, and the distribution of collagen fibers in the irradiated lung. METHODS: Thirty two C57BL mice were randomly assigned in two groups: GI (non irradiated animals) and GII (irradiated animals). The mice from GII received a single whole ¾ body radiation dose of 7Gy, using a 60Co source at a dose rate of 0.97 Gy/min. They were sacrificed by cervical dislocation at 1, 14, 30 and 90 days after radiation. RESULTS: The irradiated lungs showed: 1) vascular congestion and thickness of the alveolar septa 30 days and more intense 90 days after irradiation; 2) significant increase of collagen deposition in all time periods after irradiation; 3) weak latent TGFbeta1 activation 1 day and strong activation 14 days after irradiation in the bronchi and alveoli. Our results suggest that some bronchial and alveolar cells may have a role in the complex process of radiation-induced lung fibrosis acting as cellular sources of active TGFbeta.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:14:36Z No. of bitstreams: 1 WOS000178973900001.pdf: 373408 bytes, checksum: 88358f4e7be322ce28305b711a4625e6 (MD5) Made available in DSpace on 2014-02-26T17:14:36Z (GMT). No. of bitstreams: 1 WOS000178973900001.pdf: 373408 bytes, checksum: 88358f4e7be322ce28305b711a4625e6 (MD5) Previous issue date: 2002-11-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:24:39Z No. of bitstreams: 1 WOS000178973900001.pdf: 373408 bytes, checksum: 88358f4e7be322ce28305b711a4625e6 (MD5) Made available in DSpace on 2014-05-20T15:24:39Z (GMT). No. of bitstreams: 1 WOS000178973900001.pdf: 373408 bytes, checksum: 88358f4e7be322ce28305b711a4625e6 (MD5) Previous issue date: 2002-11-01 Experiments were undertaken to assess the role of amifostine in the activation of latent TGFbeta1 and in the smad proteins cascade (smad 2/3, smad4, smad7), focusing on megakaryocytes, in the bone marrow irradiated in vivo. Non-irradiated megakaryocytes were negative for active TGFbeta1. Immunopositivity to active TGFbeta1 was detected in megakaryocytes 10 days after irradiation in amifostine- treated and untreated marrows. Smad 2/3 and smad 4 were strongly positive in the nucleus of megakaryocytes 10 days after irradiation. At the same time, a predominant hypocellular bone marrow with foci of hematopoiesis was observed with few megakaryocytes. An increase in the number of reticulin fibers was also seen. In amifostine-treated marrows, smad 2/3 and smad4 were not detected in the nucleus but were positive in the cytoplasm of megakaryocytes 10 days after irradiation. Coincidentally, bone marrows were cellular with megakaryocytes. Smad7 immunoexpression was detected in the cytoplasm of megakaryocytes in the non-irradiated, amifostine-treated and in the irradiated, amifostine-treated marrows. Data indicate that amifostine does not prevent latent TGFbeta1 activation in irradiated megakaryocytes. While TGFbeta1 signal transduction occurs in megakaryocytes in untreated bone marrows, it is inhibited in megakaryocytes in amifostine-treated marrows due to the induction of smad 7 activation. This is the first report showing smad 7 activation by amifostine. Our results also suggest a role for TGFbeta1 as an inhibitor of megakaryocytes in vivo. (C) 2002 Wiley-Liss, Inc. Univ Fed São Paulo, Dept Med, Div Radiotherapy, São Paulo, Brazil Univ Fed São Paulo, Dept Morphol, São Paulo, Brazil Univ Fed São Paulo, Dept Pathol, São Paulo, Brazil State Univ São Paulo, Dept Histol, São Paulo, Brazil Univ Fed São Paulo, Dept Biophys, São Paulo, Brazil State Univ São Paulo, Dept Histol, São Paulo, Brazil

Granulocytes of the epigonal and Leydig organs of Rhizoprionodon lalandii were identified and classified into three different cell types, type I and type II eosinophils and neutrophils. The development of these cells in the haematopoietic tissues was dynamic, demonstrated by nuclear immunopositivity for the proliferating cell nuclear antigen (PCNA) proteins and was regulated by various cytokines, including the transforming growth factor β-1 (TGF/β 1 ). The expression pattern of these cells was heterogeneous among individual cells and TGFβ 1 -immunostaining was found principally in the cytoplasm of immature granulocytes. The presenceof TGFβ 1 in cells about to divide was demonstrated suggesting that modulation of differentiation and proliferation occurs in the haematopoietic tissues of this species of elasmobranch.

Toads have a pair of parotoid macroglands behind the eyes that secrete poison used in passive defence against predators. These macroglands are composed of juxtaposed alveoli, each one bearing a syncytial gland, all connected to the exterior by ducts. When the parotoids are bitten, the poison is expelled on the predator oral mucosa in the form of jets, causing several pharmacological actions. After poison release, the empty secretory syncytia immediately collapse in the interior of their respective alveoli and gradually start refilling. After parotoid manual compression, simulating a predator's bite, we studied, by means of morphological methods, the replacement of the poison inside the alveoli. The results showed that after compression, a considerable number of alveoli remained intact. In the alveoli that were effectively affected the recovery occurs in different levels, from total to punctual and often restrict to some areas of the syncytia. The severely affected alveoli seem not recover their original functional state. The fact that only a part of the parotoid alveoli is compressed during an attack seems to be crucial for toad survival, since the amphibian, after being bitten by a predator, do not lose all its poison stock, remaining protected in case of new attacks.

The cytochemical, immunocytochemical and ultrastructural characteristics of leukocytes and thrombocytes in the peripheral blood of the fat snook (Centropomus paralellus) - a fish occurring in Brazil - were investigated. The cytochemical methods were performed to demonstrate four enzymatic reactions - o-toluidine-hydrogen peroxide, naphtol AS-MX phosphate, naphtol AS-BI phosphate and alpha-naphtil acetate to detect myeloperoxidase (MPO), alkaline phosphatase (ALP), acid phosphatase (ACP) and non-specific esterase (α-NAE), respectively - and two non-enzymatic ones - Periodic-Acid Schiff (PAS) and Sudan black B (SBB) to detect the occurrence of glycogen and phospholipids, respectively. Immunocytochemical method utilizing polyclonal rabbit antibody against mammal metalloproteinases (MMPs) 2 and 9 were done. Standard method for Electron Microscopy (EM) was applied for the ultrastructural study. The cytochemical reactions were positive in neutrophils for MPO, ACP, α-NAE, glycogen and phospholipids; in lymphocytes for ACP and α-NAE; in monocytes for ACP and α-NAE and in thrombocytes for ACP, α-NAE and glycogen. Only neutrophils were positive for MMPs 2 and 9, and none of the cells studied were positive for ALP. Ultrastructurally: 1) neutrophil showed a spherical shape with a spherical, indented or lobulated euchromatic nucleus, and cytoplasm containing granules of varied sizes and mitochondria of varied shapes and sizes. The nucleus/cytoplasm relation and the size of granules suggest neutrophil maturation in peripheral blood; 2) lymphocytes showed partially heterochromatic nucleus and minimal cytoplasm; 3) monocytes had long cytoplasmic projections, an indented nucleus, evident nucleolus and cytoplasm with granules of varied sizes and vacuoles; 4) thrombocytes were predominantly elliptical or roughly spherical in shape, had a partially heterochromatic nucleus and cytoplasm containing electron-dense granules, intricate canalicular system and vacuoles occasionally holding phagocytic material.

Female Pipa carvalhoi incubate their eggs in the skin of the dorsum where the embryos develop until they emerge. Behavioral and morphological aspects of this reproductive mode were studied through courtship until the tadpoles emerged. Samples of the female skin were collected beginning a few hours after egg deposition and through subsequent phases of larval development and examined using standard histology, histochemistry and immunohistochemistry. The females' dorsal skin structure changes during egg implantation and development. These changes may be mediated by hormones and enzymes in a manner paralleling that of trophoblast implantation in mammal endometrium. The lack of vittelum and the extensive vascularization in the female after the 14th day of egg implantation suggest interaction through blood between the female and embryos. This hypothesis could be better accessed, if comparisons could be made with similar species such as Pipa pipa and Pipa arrabali, in which the embryos hatch only after...

Introdução: A análise morfométrica dos parâmetros nucleares, tamanho, forma e aspectos da cromatina, mostra-se útil para diagnosticar tumores malignos. O aspirado citológico coletado por agulha fina produz esfregaços suficientemente delgados para a determinação de dimensões nucleares facilmente. Objetivo: Comparar os aspectos morfométricos dos núcleos das células do aspirado proveniente de nódulos de mama. Método: Esfregaços do aspirado provenientes de 71 biópsias de nódulos de mama e determinados como benignos ou malignos, coletados por agulha fina, foram submetidos à coloração de Feulgen e as suas imagens digitais analisadas utilizando-se a Imagilab® 2,3 (Softium Informatica Ltda.), ambientado no sistema operacional WindowsTM 98 SE. Resultados: Dentre os nódulos estudados, 20 foram diagnosticados como benignos e 51 como malignos (8 GI, 23 GII e 20 GIII de acordo com o Sistema de Classificação de Guilford e Robinson). A análise morfométrica da coloração de Feulgen determinou um valor médio para os nódulos benignos de 101,6 e valor médio de 114,3 para os nódulos malignos, respectivamente. A análise morfométrica da coloração de Papanicolaou determinou um valor médio de 103,5 para GI, 131,7 para GII e 98,4 para GIII, respectivamente. Houve uma diferença estatisticamente significante entre o valor médio morfométrico de nódulos benignos e malignos e valor médio entre GI e GII, GII e GIII, mas não entre GI e GIII. Conclusões: A análise morfométrica nuclear dos aspirados de nódulo de mama é um método simples, confiável e complementar, que permitiu distinguir as lesões benignas das lesões malignas da mama.

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) O objetivo deste estudo foi determinar os parâmetros eritrocíticos, as contagens de trombócitos e de leucócitos de espécimes de Brycon amazonicus (matrinxã), criados em cativeiro e compará-los com aqueles descritos na literatura para outras espécies de Bryconinae. Além disso, foi ainda investigada a presença de células granulocíticas especiais nestes peixes. Os resultados dos parâmetros sangüíneos apresentados para B. amazonicus podem ajudar a entender melhor as características sangüíneas em população natural desta espécie de grande importância para a aqüicultura brasileira. Os parâmetros sangüíneos das espécies de Bryconinae investigadas apresentaram variação interespecíficas principalmente a contagem de eritrócitos, hemoglobina, hematócrito e volume corpuscular médio (VCM). A presença dos granulócitos sangüíneos, neutrófilos e heterófilos em matrinxã sugere que esta pode ser uma característica das espécies pertencentes à subfamília Bryconinae. Além disso, indica que a existência de células granulocíticas especiais no sangue de espécies de Bryconinae da literatura é um artefato, e isto foi aqui discutido. The aim of this study was not only to determine the red blood cells parameters, thrombocyte and leukocyte counts in farmed Brycon amazonicus (matrinxã), to compare these parameters among Bryconinae species from literature, and also to investigate the presence of special granulocytic cells in these fish. The results of the blood cells parameters here established for farmed B. amazonicus, a species of great economic importance in Brazilian aquaculture, could help a better understanding of the blood features in natural populations of this Amazon species. Blood parameters varied between Bryconinae species investigated, mainly the red blood cell counts, hemoglobin, hematocrit and mean corpuscular volume (MCV). The presence of the blood granulocytes, neutrophils and heterophils in matrinxã suggest that both leukocytes can be a characteristic for Bryconinae family. Furthermore, it indicates that the existence of special granulocytic cells in the blood of Bryconinae species from literature is an artifact, and this was herein discussed. EMBRAPA AMAPÁ Instituto Nacional de Pesquisa da Amazônia Coordenação de Pesquisa em Aqüicultura Universidade Federal do Amazonas Instituto de Ciências Biológicas Departamento de Ciências Fisiológicas UNIFESP UNIFESP, EPM, São Paulo SciELO

Objective Autoantibodies to DNA topoisomerase I (topo I) are associated with diffuse systemic sclerosis (SSc), appear to be antigen driven, and may be triggered by cryptic epitopes exposed during in vivo topo I fragmentation. These autoantibodies recognize topo I and fragments of this autoantigen generated during apoptosis and necrosis. We undertook this study to determine whether lysosomal cathepsins are involved in topo I fragmentation during necrosis. Methods Topo I cleavage during necrosis was assessed by immunoblotting of lysates from L929 fibroblasts exposed to tumor necrosis factor α (TNFα) and the broad caspase inhibitor Z-VAD-FMK, and by immunoblotting of lysates from endothelial cells treated with HgCl2. Purified topo I and L929 nuclei were incubated with cathepsins B, D, G, H, and L, and topo I cleavage was detected by immunoblotting. The intracellular localization of cathepsin L activity and topo I in necrotic cells was examined using fluorescence microscopy. Results Treatment of L929 cells with TNFα and Z-VAD-FMK induced caspase-independent cell death with necrotic morphology. This cell death involved topo I cleavage into fragments of approximately 70 kd and 45 kd. This cleavage profile was reproduced in vitro by cathepsins L and H and was inhibited by the cathepsin L inhibitor Z-FY-CHO. During necrosis, cathepsin L activity diffused from lysosomes into the cytoplasm and nucleus, whereas topo I partially relocalized to the cytoplasm. Z-FY-CHO delayed necrosis and partially blocked topo I cleavage. The topo I cleavage fragments were also detected in necrotic endothelial cells and recognized by SSc sera containing anti–topo I antibodies. Conclusion These results implicate cathepsins, particularly cathepsin L, in the cleavage of topo I during necrosis. This cleavage may generate potentially immunogenic fragments that could trigger anti–topo I immune responses in SSc.

assess the latent and active TGFb1 localization in the lung, whether or not radiation induces latent TGFbeta1 activation, and the distribution of collagen fibers in the irradiated lung.Thirty two C57BL mice were randomly assigned in two groups: GI (non irradiated animals) and GII (irradiated animals). The mice from GII received a single whole - body radiation dose of 7Gy, using a 60Co source at a dose rate of 0.97 Gy/min. They were sacrificed by cervical dislocation at 1, 14, 30 and 90 days after radiation.The irradiated lungs showed: 1) vascular congestion and thickness of the alveolar septa 30 days and more intense 90 days after irradiation; 2) significant increase of collagen deposition in all time periods after irradiation; 3) weak latent TGFbeta1 activation 1 day and strong activation 14 days after irradiation in the bronchi and alveoli. Our results suggest that some bronchial and alveolar cells may have a role in the complex process of radiation-induced lung fibrosis acting as cellular sources of active TGFbeta.

Basic hematological information about the avian in its natural habitat is limited. In this study we aimed at investigating, in the peripherical blood of the roadside hawk, Buteo magnirostris, the number of thrombocytes and their morphological, cytochemical and ultrastructural aspects. Blood smears were submited to the following methods: Leishman for morphological analysis and specific methods for the characterization of four kinds of macromolecules of different chemical nature: Glycogen, lysosomal enzymes (myeloperoxidase and the acid phosphatase), basic polyaminoacid rich proteins and phospholipids. The average number of thrombocytes was 26.8 x 10³ ± 10,2 / µL. Structurally the thrombocytes present predominantly elliptic shape, with dense and clumped nuclear chromatin. The cytoplasm is clear, vacuolated and may contain one or two purple azurophilic granules at the poles. The thrombocytes presented positive cytochemical reaction to glycogen, polyaminoacid and for acid phosphatase. Ultrastructurally, the thrombocytes appear as elliptical or spherical cells containing Golgi, much smooth endoplasmic reticulum, small mitochondria, dense osmiophilic granules, microtubules at the outermost zone of the cell and large vacuoles connected to the canalicular system or opened to the surface La información hematológica básica sobre las aves en su habitat natural es limitado. Investigamos la sangre periférica de Buteo magnirostris, el número de trombocitos y sus aspectos morfológicos, citoquímicos y ultraestructurales. Muestras de sangre fueron tratadas con los siguientes métodos: Leishman para análisis morfológico y métodos específicos para la caracterización de cuatro tipos de macromoléculas de diferente naturaleza química: Glicógeno, enzimas lisosomales (mieloperoxidasa y fosfatasa ácida), poliaminoácidos básicos ricos en proteínas y fosfolípidos. El número promedio de trombocitos fue 26.8 x 10³ ± 10,2 / mL. Estructuralmente los trombocitos presentaban predominantemente forma elíptica, con densa y agrupada cromatina nuclear. El citoplasma era claro, vacuolado y contenía uno o dos gránulos azurofílicos morados en los polos. Los trombocitos presentaron reacción citoquímica positiva para glucógeno, poliaminoácidos y fosfatasa ácida. Ultraestructuralmente, los trombocitos se presentaban como células elípticas o esféricas conteniendo Golgi, con mucho retículo endoplasmático liso, pequeñas mitocondrias, densos gránulos osmiofílicos, microtúbulos en la zona exterior de la célula y vacuolas grandes conectadas a través del sistema canalicular o abiertas a la superficie

OBJETIVO: Investigar no pulmão, por imunohistoquímica, a localização das proteínas TGFbeta1 latente e TGFbeta1 ativa, se ocorre ativação radioinduzida da proteína TGFbeta1 latente e a distribuição das fibras colágenas em diversos períodos de tempo após irradiação. MÉTODOS: 32 camundongos isogênicos (C57BL) foram divididos em dois grupos: GI (não irradiado) com 12 animais e GII (irradiado) com 20 animais. Os animais do GII receberam radiação gama (telecobaltoterapia, 60Co, com rendimento de 0,97Gy/min, dose única de 7Gy em corpo inteiro). Os camundongos dos grupos I e II foram sacrificados por estiramento cervical nos períodos de 1, 14, 30 e 90 dias após irradiação. RESULTADOS: Os pulmões irradiados apresentaram: 1) congestão vascular e espessamento dos septos alveolares aos 30 dias e mais intensamente aos 90 dias depois da irradiação; 2) aumento significante da deposição de colágeno em todos os períodos de tempo após irradiação; 3) fraca ativação da proteína TGFbeta1 latente em um dia e intensa aos 14 dias depois da irradiação em brônquios e alvéolos. Nossos resultados sugerem que células dos brônquios e alvéolos podem participar do complexo mecanismo de fibrose pulmonar radioinduzida atuando como fontes da proteína TGFbeta1 ativa.

Morfologicamente foram identificados no sangue de Oreochromis niloticus sete tipos de células: eritrócitos, trombócitos, neutrófilos, eosinófilos, basófilos, linfócitos e monócitos. Em relação aos resultados citoquímicos foi contastada a presença de glicogênio em neutrófilos, trombócitos e em alguns linfócitos e monócitos. Os grânulos citoplasmáticos de neutrófilos e eosinófilos mostraram positividade para mieloperoxidade e Sudan black. O azul de bromofenol foi totalmente positivo em eritrócitos e eosinófilos. Morphologicaly, seven types of cells were identified in the blood of Oreochromis niloticus: erythrocytes, thrombocytes, neutrophils, eosinophils, basophils, lymphocytes and monocytes. Glycogen was present not only in the cytoplasm of neutrophils and thrombocytes but also in some lymphocytes and monocytes. The positive reaction for myeloperoxidase and Sudan black was observed in neutrophils and eosinophils. The bromphenol blue method was strongly positive for erythrocytes and eosinophils. Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de Morfologia Universidade de São Paulo Faculdade de Medicina Veterinária Departamento de Patologia UNIFESP, EPM, Depto. de Morfologia SciELO

Las caracteristicas morfologicas de sangre periferica del pez Salminus maxillosus fueron descritas usando microscopia de luz. Para demostrar el comportamiento citoquimico de las celulas sanguineas del pez en estudio, fueron aplicados metodos especificos para la deteccion de cuatro tipos basicos de componentes macromoleculares, de naturaleza quimica diferente: carbohidratos, lipidos, proteinas enzimaticas y estructurales. Fueron observados 6 tipos de celulas: eritrocitos, trombocitos, monocitos, linfocitos, neutrofilos de los tipos I y II y eosinofilos. Los trombocitos y los neutrofilos de los tipos I y II presentaron glucogeno en el citoplasma. Los neutrofilos del tipo II presentan granulos sudanofilos y mieloperoxidasa. Ademas presentan, reaccion difusa intensa para proteinas en el citoplasma. Los eosinofilos muestran reaccion intensa en sus granulos.

Foram utilizados 10 jacarés Caiman crocodilus yacare, colhendo-se 5 ml do sangue periférico de cada animal. A análise morfológica foi realizada após coloração 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 são 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 salmão, 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 relação 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 projeções e grânulos azurófilos. Monócito azurófilo, de formato esférico, núcleo excêntrico e citoplasma basófilo contendo grânulos azurófilos abundantes. Samples of peripheral blood were collected from 10 healthy Caiman crocodilus yacare and stained with Leishman, for morphological analysis, or submitted to the following cytochemical methods: PAS, Sudan black B, o-toluidine and mercury bromophenol blue. Seven types of cells were identified: erythrocytes, thrombocytes, heterophils, eosinophils, basophils, lymphocytes and azurofilic monocytes. Erythrocytes were elliptical and positive for bromophenol blue. Some mitotic figures were noted. Spindle-shaped thrombocytes showed large nucleus with striking indentation. The cytoplasm was attenuated at one or both poles into knob-like projections containing glycogen. Heterophils revealed round eccentric nuclei and cytoplasm containing glycogen and many polymorphic granules. These granules showed partial positive reaction for Sudan and activity for myeloperoxidase. Eosinophils showed eccentrically located lenticular-shaped nuclei and numerous round granules stained positive with myeloperoxidase and Sudan. Basophils revealed a round central nuclei and cytoplasm with few basophilic round granules. Lymphocytes appeared as polygonal or irregular cells with thin cytoplasm forming bleb-like protrusions. Azurophilic monocytes were round with eccentrically located nuclei and basophilic cytoplasm containing numerous azurophilic granules.

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 siaIidase 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 alpha 2-->3-linked sialic acids at pH 4-8, with maximaI 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 (K-i = 0.075 mM) and differently from many sialidases, with exception of Salmonella typhimurium sialidase, it is inhibited competitively by HEPES (K-i = 15 mM). the fact that T. infestans sialidase is released with the saliva and can hydrolyze sialyl-Lewis(x) blood groups, which are the ligands for selectins, suggests that it might have a role in the blood feeding. Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Histol, BR-04023062 São Paulo, Brazil Ludwig Inst Canc Res, BR-01509010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Histol, BR-04023062 São Paulo, Brazil Web of Science

RESUMEN: Muestras de sangre periférica de reptiles, jóvenes y sanos, fueron recolectadas del seno venoso cervical, en tubos que contenían EDTA 10%. Inmediatamente los frotis fueron sometidos a los siguientes procedimientos: a) Tinción con solución de Rosenfeld para el análisis morfológico. b)Detección de la fosfatasa alcalina con naftol AS-MX fosfato y mieloperoxidasa (pH 7,0 y pH10,0) con o-toluidina H(2)0(2). Una parte de la sangre con EDTA fue centrifugada por 15' a 2.000rpm y el coágulo conteniendo los leucocitos, fue dividido en dos fragmentos y tratados con: a) glutaraldehído a 2% (1h a 4C) para la reacción de la fosfatasa alcalina; b)glutaraldehído a 1,25% + paraformaldehído al 1% (1h 4C), para la reacción de la mieloperoxidasa. Los eosinófilos revelaron núcleo excéntrico redondeado, oval o lobulado con nucléolo prominente. El citoplasma de los eosinófilos es ocupado por gránulos eosinofílicos evidentes (grandes, aproximadamente esféricos, compactos, uniformes en tamaño). No fue observada actividad de la fosfatasa alcalina en eosinófilos. Una reacción positiva débil en pH 7,0 y fuerte en pH 10,0, para mieloperoxidasa fue visualizada como un precipitado marrón, en los eosinófilos. El microscopio electrónico reveló actividad de la mieloperoxidasa solamente en algunos gránulos en pH 7,0 y en la mayoría de los gránulos en pH 10,0. Proteínas básicas produjeron reacción visible, principalmente en la periferia de todos los gránulos citoplasmáticos de los eosinófilos. SUMMARY: Samples of peripheral blood from healthy young reptiles were collected from the cervical venous sinus in tubes containing EDTA 10%. Freshly prepared smears were used for the following purpuses: a) Staining in Rosenfeld solution for the morphological analyses. b) Detection of alkaline phosphatase with naphthol ASMX-phosphate and myeloperoxidase (pH 7,0 and pH10,0) with o-toluidine H(2)0(2). The same blood samples with EDTA were centrifuged for 15' at 2,000rpm and the buffy coat containing leukocytes was devided in two fragments and one of them treated with glutaraldehyde (1h at 4C) and used for alkaline phosphatase reaction and the other 1,25% glutaraldehyde + 1% paraformaldehyde (1h 4C) and used for myeloperoxidase reaction. The eosinophils revealed round, oval or lobed eccentric nuclei with prominent nucleoli. The cytoplasm of eosinophils is dominated by striking eosinophilic granules (large, nearly spherical, closely packed, uniform in size). No alkaline phosphatase activity was observed in the eosinophils. A positive reaction for myeloperoxidase (faint at ph 7,0 an strong at pH 10,0) was visualized as a brown precipitate in the eosinophils. EM observation revealed myeloperoxidase activity only in a few granules (at pH 7,0) and in a majority of the granules (at pH 10,0). Basic protein reaction product is visible essentially at the periphery of all the citoplasmic granules in the eosinophils.

Hemogram and cell morphology of peripheric blood samples obtained from 10 Oreochromis niloticus fishes have been evaluated. Some of these samples have been used in the accomplishment of hematimetry: eritrocytes count (x = 2.35 x 106/mm3 ±0.12), hemoblobin rate ( x = 7.04 g/dL ± 0.38), hematocrit (x = 27.85% ± 1.62), MHC (x = 30.31 pg ± 1.64), MCV (x = 118.6 p3 ± 3.53), MCHC (x = 25.78% ± 1.72), total leucocytes count (x = 16.08 x 103/mm3 ± 1.28), trombocytes count ( x = 61.69 x 103/mm3 ± 5.96). Morphological evaluation of blood cells was performed by means blood smears extension submitted to Wright staining. Both nucleated eritrocytes and trombocytes presented an elliptical shape; neutrophiles and eosinophiles were spherical with spherical nucleus presenting different sizes, they also showed profuse citoplasms which were slightly basophilic and highly acidophilous, respectively. Polimorphonuclear shapes in these cells have not been verified. Basophiles have shown spherical nucleus and metachromic granulations. Monocytes as well as lymphocytes resembled morphologically those of mammals. 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 106/mm3 ± 0,12), taxa de hemoglobina, (x = 7,04 g/dL ± 0,38), hematócrito (x = 27,85% ± 1,62), HCM (x = 30,31pg ± 1,64), VCM (x = 118,6p3 ± 3,53), CHCM (x = 25,78% ± 1,72), contagem global de leucócitos (x = 16,08 x 103/mm3 ± 1,28), contagem de trombócitos (x = 61,69 x 103/mm3 ± 5,96), sendo a avaliação da morfologia das células do sangue reallizada através do estudo de extensões sangüíneas submetidas à coloração 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. Não foram vistas as formas polimorfonucleares dessas células. Os basófilos apresentaram núcleo esférico e granuiações metacromáticas. Os monócitos e os linfócitos assemelharam-se morfologicamente aos dos mamíferos.

Fueron utilizados 10 ejemplares de Caiman crocodilus yacare provenientes del municipio de Cáceres (Estado de Mato Grosso do Sul, Brasil), a cada uno de los cuales les fueron extraidos 5ml de sangre periférica. Las muestras obtenidas fueron utilizadas para análisis ultraestructurales, referentes a trombocitos, eosinófilos y heterófilos. Los trombocitos son de forma elíptica, poseen citoplasma abundante sólo en los polos y su núcleo presenta surcos profundos o hendiduras acentuadas, mostrando algunos poros en su envoltura. Su citoplasma contiene abundantes túbulos y vesículas electrolúcidas del sistema canalicular. Son observados vacúolas con cuerpos densos y microtúbulos. Los heterófilos poseen formato aproximadamente esférico y núcleo excéntrico o periférico. Su citoplasma está repleto de gránulos polimórficos de formas de huso, en baqueta, oval o esférica, con superficie, en general, irregular. Los eosinófilos son esféricos y de núcleo lenticular excéntrico o periférico. El citoplasma contiene gránulos esféricos u ovales. Cabe destacar la presencia de gránulos citoplasmáticos voluminosos con diferencias en la electrodensidad, de superficie uniforme, pudiendo contener una estructura interna heterogénea. En heterófilos y eosinófilos, el núcleo presenta heterocromatina, eucromatina, nucléolo y la envoltura nuclear es porosa Little information is available on the ultrastructural characteristics of the blood cells in reptiles. In the present study we have examined the fine structure of heterophils, eosinophils and thrombocytes from the blood of a crocodile (Caiman crocodilus yacare). Samples of peripheral blood from healthy young were collected with EDTA and centrifuged. The buff coat was collected and fixed in half strength Karnovsky solution. Spindl-eshaped thrombocytes showed large nuclei with prominent heterochromatin juxtaposed to the nuclear envelop. Cytoplasmic organelles were scarce: there were mitochondria, some profiles of roughsurface endoplasmic reticulum, free ribosomes and microtubules. Many tubular structures and vacuoles were distributed throughout the cytoplasm, forming the open canalicular membrane system. Heterophils and eosinphils revealed round eccentric nuclei with prominent nucleoli. The cytoplasm of heterophils contained many polymorphic granules with homogeneous electrondense matrix. Ribosomes, roug-surfaced endoplasmic reticulum and many clear vesicles were observed. Some fibrillar structures were also present. In addition to the usual organelles, the cytoplasm of eosinophils were essentially filled with many round or slightly oval granules with heterogeneous electron-dense matrix; in some of them a fine granular structure was visible. Both heterophils and eosinophils showed long numerous pseudopodia

Taking into account the great sensitivity of the intestinal mucosa to X irradiation we aimed at investigating the radioprotection afforded by the S-2-(3-aminopropylamino) ethylphosphorothioic acid (WR2721) on the mice rectum exposed to a single whole body X ray dose of 7 Gy and sacrificed at 1, 3, 6 and 10 days after irradiation. Comparing the data of protected group with those of the unprotected one the following features were observed in the first one: statistically significant increase of both crypt cells per crypt (CrC/Cr) at 1, 3, 6 and 10 days after irradiation and of mitotic figures per crypt (MF/Cr) at 3, 6 and 10 days post exposure. In this way it became clear that these morphometric parameters allowed the estimation of WR2721 rectum protection through the crypt cellularity and epithelial cell kinetics En base al elevado grado de radio-sensibilidad de la mucosa intestinal, ha sido estudiada la acción radioprotectora del acido S2(3aminopropilamino) etilfosforotioico (WR2721) sobre el epitelio de recto de ratones expuestos cuerpo entereo a los rayos X, dosis única de 7 Gy, y sacrificados después de 1, 3, 6 y 10 días. Al comparar las muestras de segmentos del recto de animales protegidos y no protegidos, fue observado, de manera general, que en los protegidos, ha ocurrido aumento estadísticamente significativo en el número de células de la crypta por crypta (CrC/Cr) después de 1, 3, 6 y 10 días luego de la irradiacón, y en el número de figuras de mitosis por crypta (MF/Cr) después de 3, 6 y 10 días. Se puede concluir que los parámetros morfométricos referidos, permitieron estimar cuantitativamente la radioprotección por el WR2721 en el recto de los ratones, evaluada por la celularidad de la cripta y por la cinética de las células epiteliales intestinales

Amostras de sangue periférico foram obtidas de cinco primatas Cebus apella (macaco prego), antes e após exposição 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 realização de extensões sanguíneas e submetidas à coloração de Wright c aos seguintes métodos citoquímicos: ácido periódico Schiff (PAS) para a identificação de glicogênio, Sudan black B para a detecção de fosfolipídios e o-toluidina-água oxigenada para a demonstração da mieloperoxidase. Com o método de Wright foram observadas aos 1, 3 e 6 dias após exposição à radiação, alterações nucleares e citoplasmáticas. Após aplicação dos métodos citoquímicos acima referidos, foram observados, de modo geral, nos mesmos períodos de tempo após irradiação: alteração no padrão de distribuição 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 3º. dia após irradiação. Noventa dias após, não foi observada nenhuma alteração acima referida. The effects of ionizing radiation on the neutrophils of Primate Cebus apella were studied after whole-body x-radiation to a single exposure of 25.8 mC/kg (100R), Wright's stained preparations showed changes in the nucleus and the cytoplasm of neutrophils at 1, 3 and 6 days after irradiation. During this period of time, the cytochemical methods revealed a considerable variation in the pattern of distribution of glycogen, sudanophilic and myeloperoxidase positive granules. Under these same experimental conditions the number of caryoschizes increased on the first and third day. On the ninetieth day post exposure, the morphological and cytochemical appearances of neutrophils as well as the number of caryoschizes were similar to the controls.