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Made available in DSpace on 2021-07-14T10:42:32Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-05. Added 1 bitstream(s) on 2021-07-14T11:36:27Z : No. of bitstreams: 1 S0100-736X2021000100509.pdf: 675127 bytes, checksum: 8557958b3f0c8123c89ac10d71a538ad (MD5) Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos. This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots. Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias

Leukoreduction (LR) is a technique that consists of reducing the number of leukocytes in whole blood or blood components that can contribute to decreasing storage lesions and the occurrence of post-transfusion complications. We propose that using a blood bag with pre-storage leukocyte filtration is sufficient for blood conservation under field conditions. Ten healthy Nelore cows were used. Whole blood was sampled from each animal and stored at 2 to 6 &deg, C in CPD/SAG-M (citrate phosphate dextrose bag with a saline, adenine, glucose, mannitol satellite bag) triple bags (Control) and in CPD/SAG-M quadruple bags with a leukocyte filter (Filter). At baseline and after 7, 14, 21, 28, 35, and 42 days (D0, D7, D14, D21, D28, D35, and D42, respectively), complete hematological, blood gas, and biochemical evaluations were determined. The filtered bag removed 99.3% of white blood cells from cattle blood, and the entire filtration process was performed in the field. There was a reduction in the number of red blood cells (RBCs) in both groups from D14 onward, with a decrease of 19.7% and 17.1% at D42 for the Control and Filter bags, respectively. The hemoglobin (Hb) concentration had variation in both groups. Potassium, pO2, pCO2, and sO2 increased, and sodium, bicarbonate, and pH decreased during storage. The filtered bag was efficient in removing white cells from cattle whole blood and could be used under field conditions. Blood stored after LR showed differences (p &lt, 0.05) in blood gas analysis towards a better quality of stored blood (e.g., higher pH, lower pCO2, higher sO2). Further experimental studies are required to prove that blood without white cells results in a decrease in transfusion reactions in cattle.

Immune functions in teleosts are straightly influenced by environmental temperature. Low temperatures usually affect negatively the immune status of fishes favoring the incidence of diseases. Immune cells in fishes are protected from peroxidation by dietary vitamin E. Innate immune defenses of Piaractus mesopotamicus were evaluated in fish fed with diets supplemented with vitamin E for 45 days. The fish endured decreases in environmental fluctuations of temperature for 24 and 72 h. The most effects were observed at the 24th hour, in which the level of plasma cortisol increased independently of dietary supplement of vitamin E and a lessening of MNL and a raise of GL counting were observed in fish fed with supplemented diets. The lowest SL variation was observed in fish fed with the maximum level of vitamin E, but the dietary vitamin E decreased the HAC. However, the stressing condition increased HAC. Dietary vitamin E used in the present experimental conditions did not present remarkable effects on the immune frame of pacu to cope with environmental thermal fluctuations but other investigations concerning the vitamin half-life and its association with unsaturated dietary lipids seem to be fundamental.

RESUMO Aparelhos de automação, cujo uso está estabelecido na hematologia dos animais domésticos, ainda não são empregados rotineiramente em felídeos selvagens. Esse estudo objetivou avaliar a técnica automatizada na contagem de células sanguíneas de jaguatiricas (Leopardus pardalis), comparando-a com a técnica manual. Foram coletadas amostras sanguíneas de oito jaguatiricas, que se submeteram à contagem de eritrócitos, leucócitos, plaquetas e à determinação do volume globular pelo método manual padrão e por meio do aparelho BC - 2800 VET® com a configuração para gatos domésticos. Os resultados foram avaliados por meio do teste t para dados pareados, e as técnicas submetidas à análise de correlação de Pearson. A técnica automatizada demonstrou resultados estatisticamente semelhantes para eritrócitos e leucócitos. Apenas para volume globular houve diferença significativa entre as técnicas manual e automatizada, mas a correlação foi alta. Apesar de não haver diferença significativa entre as técnicas para plaquetas, a correlação foi baixa. Conclui-se que o aparelho BC - 2800 VET® com a configuração para gatos domésticos é uma técnica confiável na realização do eritrograma e do leucograma para jaguatiricas. Para a determinação do parâmetro volume globular, o aparelho pode ser utilizado, desde que se faça a correção. Para a contagem de plaquetas, a técnica manual é recomendada. ABSTRACT Automated equipment, whose use is established in hematology of domestic animals, is not yet routinely used in blood cells count of wildlife, due to lack of studies that validate its use. The purpose of this study was to evaluate the automated technique for blood cells count of ocelots (Leopardus pardalis), comparing it with the manual technique. Blood samples were collected from eight ocelots, which were submitted to counting of erythrocytes, leukocytes, platelets and packed cell volume by the standard manual method and by the device VET ® 2800 BC with the configuration for domestic cats. The results were evaluated using the t test for paired data and the techniques submitted to Pearson correlation. The automated technique showed statistically similar results to erythrocytes and leukocytes. Significant difference was found only for packed cell volume between the manual and automated techniques, but the correlation was high. Although there was no significant difference between the techniques for platelets, the correlation was low. We concluded that the 2800 BC VET ® device with the configuration for domestic cats is a reliable technique in performing the erythrocyte and leukocyte counts for ocelots. The device may be used to determine packed cell volume, provided the correction is made. The manual technique is recommended for the platelet count.

Introduction: Malaria in pregnancy very often includes gestational (parasites in maternal peripheral blood) and placental (parasites in placental blood) infection, but the later condition can only be detected after delivery. High frequency of placental plasmodial infection has been confirmed in many countries and is associated with negative birth outcomes. With the hypothesis that placental infection is accompanied by hemozoin circulation in maternal peripheral blood, an exploratory study was conducted to evaluate the association between peripheral leukocytes with hemozoin and placental infection by Plasmodium vivax or Plasmodium falciparum in parturient women. Methodology: A descriptive, transversal and exploratory (pilot type) study was carried out with women from two malaria-endemic localities of northwest Colombia. A total of 25 parturient women with confirmed placental infection and 25 without placental infection were included. Two independent readers measured the number of leukocytes with hemozoin in thick smears of maternal peripheral blood. Plasmodial infection in maternal peripheral blood and placental blood was detected by thick smear and quantitative polymerase chain reaction (qPCR). Results: Four parturient women had leukocytes with hemozoin in peripheral blood; three of them had placental plasmodial infection and one was negative for placental infection. No statistically significant association between leukocytes with hemozoin in peripheral blood and placental infection was observed. Conclusions: With this limited sample size, detection of leukocytes with hemozoin by thick smear of maternal peripheral blood did not indicate presence of placental infection.

The objective of this study was to evaluate food palatability, digestibility and blood parameter of dogs fed diets containing increasing levels of spray-dried porcine plasma (SDP). We also aimed to evaluate the SDP digestibility for dogs. We used 36 dogs distributed in 3 experiments. In the digestibility assay (Experiment 1), eight adult Beagles were distributed according to a double Latin square design (4 x 4). The following diets were evaluated: 0, 4, 8, and 12% SDP of diet. In the palatability trial (Experiment 2), 20 adult dogs were used. Diets containing 0 and 4% SDP were pair-wised compared. Blood parameters (Experiment 3) were evaluated in eight adult Beagles. Dogs were fed the diet with 0% for 10 days and then the diet with 12% SDP for further 10 days. Dietary SDP inclusion promoted a linear increase (P0.05). In conclusion, SDP shows high digestibility of CP and DM, being feasible its use in diets for dogs. However, it is important to study the effects of inclusion levels lower than 4%, in order to do not compromise diet palatability. O objetivo do estudo foi avaliar a digestibilidade, palatabilidade e parâmetros sanguíneos em cães alimentados com dietas contendo crescentes níveis de plasma suíno spray-dried (PSD). Ainda, objetivou-se isolar a digestibilidade do PSD em cães. Foram utilizados 36 cães distribuídos em 3 experimentos. Para o ensaio de digestibilidade (Experimento 1) foram utilizados oito cães adultos da raça Beagle, distribuídos em quadrado latino duplo (4x4). Foram avaliadas quatro dietas: 0%, 4%, 8% e 12% PSD. Para avaliação da palatabilidade (Experimento 2) foram utilizados 20 cães adultos. Foram comparadas duas dietas: 0% PSD vs. 4% PSD. Para a avaliação hematológica (Experimento 3) foram utilizados oito cães adultos Beagles. Todos os cães foram alimentados com a dieta 0% PSD por 10 dias e posteriormente com a dieta contendo 12% PSD por mais 10 dias. Houve aumento linear (P0,05), nos cães alimentados com 12% PSD. Conclui-se que o PSD apresenta elevada digestibilidade da PB e MS, sendo viável o seu uso em dietas para cães. Entretanto, é importante estudar os efeitos de níveis de inclusão menores que 4%, a fim de não comprometer a palatabilidade da dieta.

Animal studies, mostly performed in rodents, show the beneficial anti-obesity effects of cold studies. This is due to thermogenic activation of brown adipose tissue (BAT), a tissue also recently discovered in adult humans. Studies in humans, however, are hampered by the accessibility of most tissues. In contrast, peripheral blood mononuclear cells (PBMC) are accessible and share the expression profile of different sets of genes with other tissues, including those that reflect metabolic responses. Ferrets are an animal model physiologically closer to humans than rodents. Here, we investigated the effects on ferrets of one-week acclimation to 4 degrees C by analysing the PBMC transcriptome. Cold exposure deeply affected PBMC gene expression, producing a widespread down-regulation of genes involved in different biological pathways (cell cycle, gene expression regulation/protein synthesis, immune response, signal transduction, and genes related to extracellular matrix/cytoskeleton), while thermogenic and glycogenolysis-related processes were increased. Results obtained in PBMC reflected those of adipose tissue, but hardly those of the liver. Our study, using ferret as a model, reinforce PBMC usefulness as sentinel biological material for cold-exposure studies in order to deepen our understanding of the general and specific pathways affected by cold acclimation. This is relevant for future development of therapies to be used clinically., CIBER de Fisiopatologia de la Obesidad y Nutricion is an initiative of the ISCIII. Laboratory of Molecular Biology, Nutrition and Biotechnology as well as Human and Animal Physiology are members of the European Research Network of Excellence NuGO (The European Nutrigenomics Organization, EU Contract: FOOD-CT-2004-506360 NUGO). Experimental work was supported by the Spanish Government (Ministerio de Economia y Competitividad, INTERBIOBES -AGL2015-67019-P-(MINECO/FEDER, UE)) and the University of the Balearic Islands (BIOTHERM, FA42/2016). Open access publication was funded by the IdISBa Liberi program.

The objective of the study was to identify leukocyte populations in the postnatal thymus of alpacas under 60 days of age. Euthanasia was performed on six Huacaya alpacas in Huanta province, Ayacucho, Peru. Thymus samples were obtained, embedded in OCT and immediately frozen in liquid nitrogen and stored at -70 ºC until processing. Frozen sections were prepared and mounted with organosilane. Immunohistochemistry was performed with mouse primary antibody and rabbit secondary antibodies. The reaction was evidenced using the avidin-biotin-peroxidase complex and was counter-stained with hematoxylin. Specific monoclonal antibodies were used for the identification of leukocytic subpopulations of the thymus. Five visual fields were randomly evaluated at 400X. The distribution and density of the staining was classified as negative (-), inconstant (±), slight (+), moderate (++) and strong (+++). The results show that the largest population of leukocyte cells was located in the cortex of the thymus and corresponded to immature cells that do not express surface markers; likewise, the highest population of immunomarked cells, in maturation, were located in the medulla of the thymus, predominantly the TCR-αβ lymphocytes, with low values of γδ TCR lymphocytes, CD8 lymphocytes, B lymphocytes and monocytes/macrophages, with inconstant lymphocyte values CD4. In the cortex of the thymus there were low values of αβ TCR lymphocytes and CD8 lymphocytes, and inconstant values of monocytes/macrophages, without evidence of γδTCR lymphocytes, CD4 lymphocytes and B lymphocytes. El objetivo del estudio fue identificar las poblaciones leucocitarias en el timo posnatal de alpacas menores de 60 días de edad. Se realizó la eutanasia a seis alpacas Huacaya en la provincia de Huanta, Ayacucho, Perú. Se obtuvieron muestras de timo que fueron embebidas en OTC y congeladas inmediatamente en nitrógeno líquido y conservadas a -70 ºC hasta su procesamiento. Se prepararon secciones congeladas y se fijaron a la lámina con el adhesivo organosilano. Se realizó la inmunohistoquímica con anticuerpo primario de ratón y secundarios de conejo. La reacción se evidenció empleando el complejo avidina-biotina-peroxidasa y se contra-coloreó con Hematoxilina. Se emplearon anticuerpos monoclonales específicos para la identificación de las subpoblaciones leucocíticas del timo; se evaluaron cinco campos visuales de forma aleatoria a un aumento de 400X. La distribución y densidad de la tinción se clasificó como negativa (-), inconstante (±), ligera (+), moderada (++) y fuerte (+++). Los resultados muestran que la mayor población de células leucocitarias se ubicó en la corteza del timo y correspondieron a células inmaduras que no expresan marcadores de superficie; así mismo, la mayor población de células inmunomarcadas, en maduración, se ubicaron en la médula del timo, predominando los linfocitos TCR αβ, con valores bajos de linfocitos TCR γδ, linfocitos CD8, linfocitos B y monocitos/macrófagos, con valores inconstantes de linfocitos CD4. En la corteza del timo se observaron valores bajos de linfocitos TCR αβ y linfocitos CD8, y valores inconstantes de monocitos/macrófagos, sin evidencias de linfocitos TCR γδ, linfocitos CD4 y linfocitos B.

Pre-eclampsia (PE) is a hypertensive disorder that affects an important number of pregnant women worldwide. The exact causes of PE remain poorly understood. However, inflammation and deregulation of innate immune cells, such as natural killer (NK) cells, contribute to PE pathogenesis. Besides, the mother's genetic background also impacts on PE susceptibility. Thus, genetic variants that potentially modify the behaviour of inflammatory cells may help us to understand the causes of PE. Variants of genes encoding NKG2C (expressed in NK cells) and C-C chemokine receptor type 5 (CCR5) (expressed mainly in leucocytes) are important targets in the study of gestational disorders. In this context, we evaluated the impact of both NKGC2 gene deletion and CCR5Δ32 gene variant on PE susceptibility in a population sample from central-southeast Brazil composed by 369 women (156 with PE and 213 healthy pregnant women). No statistically significant association between the NKG2C gene deletion and susceptibility to PE was observed. However, taking into consideration the important role of NK cells in pregnancy, the influence of NKG2C gene deletion on PE pathogenesis should not be ruled out and deserves further studies in populations with different genetic/ethnic backgrounds. In addition, our results regarding CCR5Δ32 corroborate previous data from our group approaching a distinct cohort and reinforce CCR5Δ32 as a protective factor against PE development (p

The effect of adding Agaricus blazei to the diet of broilers on immunity, serum parameters, and antioxidant activity was evaluated. A total of 840 1-day-old chicks were used, distributed among five levels of a completely randomized design (0.0, 0.05, 0.10, 0.15, and 0.20% A. blazei), with six replications and 28 birds per experimental unit. The weights of the thymus, spleen and cloacal bursa were not influenced (P > 0.05). Leukocytes, macrophages and nitric oxide were unaffected (P > 0.05), but at each supplementation level compared with the control, differences appeared in the percentages of eosinophils and macrophages (P < 0.05) at inclusion levels of 0.10, 0.15 and 0.20%. At 28 days, the antibody titer against Newcastle disease showed a quadratic response (P < 0.05) with supplementation, and from the estimated level of 0.08% the production of antibodies was stimulated; however, the same behavior was not observed (P > 0.05) at 42 days. Hypocholesterolemic effect was demonstrated (P < 0.05), but the serum triglyceride concentration was not affected (P > 0.05). The antioxidant activity of mushroom showed a positive linear effect (P < 0.05) on DPPH capture on day zero of meat cooling. The inclusion of A. blazei in the diet of broilers provided an immunostimulatory activity and hypocholesterolemic effect. Residual compounds with antioxidant activity were present in the meat, which may promote tissue protection of the animal in vivo, making possible the use of A. blazei as a natural additive.

Euphorbia tirucalli (L.), commonly known as aveloz, has been indiscriminately used in popular medicine to treat various illnesses. However, some components can have devastating consequences. Injury to a cell's genetic material can cause mutations, cancer, and cell death. Our main goal in this work was to evaluate the genotoxic and cytotoxic effects of E. tirucalli extract on human leukocytes. For this purpose, we performed a phytochemical analysis to evaluate the plant's components. In the second step, we treated cultured human leukocytes with different concentrations of the dry extract of the plant and then evaluated the oxidative and genotoxic profiles of these leukocytes. We found that at 1% and 10% concentrations, the aveloz extract acted as a genotoxic agent that could damage DNA and increase oxidative damage. We conclude that despite its popular use, aveloz can act as a genotoxic agent, especially when it contains phorbol ester. Aveloz's indiscriminate use might actually promote tumors and therefore carry a considerable genetic risk for its users. RESUMO Euphorbia tirucalli (L.), comumente conhecida como aveloz, tem sido indiscriminadamente usada na medicina popular para o tratamento de várias doenças. Entretanto, alguns componentes podem ter consequências devastadoras. A lesão do material genético da célula pode causar mutações, câncer e morte celular. Nosso principal objetivo neste trabalho foi avaliar os efeitos genotóxicos e citotóxicos do extrato de E. tirucalli em leucócitos humanos. Para este propósito, realizamos uma análise fitoquímica dos componentes da planta. Em uma segunda etapa, culturas de leucócitos humanos foram tratados com diferentes concentrações do extrato seco da planta e então avaliamos os perfis oxidativos e genotóxicos nestes leucócitos. Nossos resultados mostraram que nas concentrações de 1% e 10%, o extrato de aveloz atua como agente genotóxico, o qual pode danificar o DNA e aumentar a dano oxidativo. Concluímos que, apesar do uso popular, a aveloz pode atuar como agente genotóxico, especialmente por conter o éster de forbol. O uso indiscriminado da aveloz pode na verdade promover tumores e, portanto, carrega um risco genético considerável para seus usuários.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior As aves são animais eficientes em responder a alterações no equilíbrio do meio ambiente, pois são organismos sensíveis a estresses. O perfil leucocitário é considerado uma ferramenta confiável na mensuração de estresse, pois tem sido relacionado a diversos fatores estressores como: reprodução, baixa disponibilidade de alimento, poluição e parasitismo. As aves são acometidas por diversa fauna parasitária, tais como os hemoparasitos, que podem comprometer a sobrevivência e as aptidões dos hospedeiros. Os objetivos desta dissertação foram examinar o perfil leucocitário e os hemoparasitos de uma população de Antilophia galeata, e verificar se há a associação de ambos a características biológicas e ecológicas da espécie. Em um fragmento florestal de Cerrado foram capturados 151 indivíduos em 13 campanhas de campo (06/2013 a 12/2016). Foram coletados os seguintes dados: sexo, faixa etária, biomassa, comprimento dos tarsos, presença/ausência de carrapatos, muda de penas e placa de incubação, além de amostras de sangue para confecção de extensões sanguíneas, as quais foram analisadas em microscópio óptico para identificação e quantificação dos leucócitos, micronúcleos e parasitos. Os dados coletados foram divididos em dois capítulos, o primeiro focado no perfil leucocitário, utilizando 89 indivíduos (capturados em 2013-2015) e o segundo, com enfoque nos hemoparasitos, com 62 indivíduos (2016). Em relação ao perfil leucocitário, o número de linfócitos foi maior nos machos, nos indivíduos em fase não reprodutiva e com carrapatos. Não houve diferença nos componentes leucocitários em relação às faixas etárias, muda de penas e condição corporal. A quantidade de micronúcleos foi relacionada positivamente com os valores totais de leucócitos, razão H/L, heterófilos, basófilos e monócitos. Em relação aos hemoparasitos, 33 (53,2%) indivíduos estavam infectados, sendo 32 (51,6%) por Plasmodium spp. e um (1,61%) por Haemoproteus spp. Não houve diferença na parasitemia entre as estações, sexos, presença e ausência de reprodução, muda de penas e ectoparasitos. Não houve correlação entre condição corporal e parasitemia. Houve correlação positiva entre parasitemia e a quantidade total de leucócitos, sugerindo que estes aumentam de acordo com a parasitemia, o que indica que os indivíduos podem estar sendo eficientes em controlar as infecções. O atual estudo evidenciou que o perfil leucocitário é útil em responder questões ecológicas, as quais podem ser aplicadas na conservação da espécie, pelas variações leucocitárias estarem relacionadas a fatores estressores como reprodução e quantidade de micronúcleos. Quanto aos hemoparasitos, os resultados sugerem que a população é tolerante a parasitemia encontrada, pois não foi detectado nenhum efeito negativo. Birds are efficient animals in responding to changes in the balance of the environment, as they are stress-sensitive organisms. The leukocyte profile is considered a reliable tool in the measurement of stress, since it is related to several stressors such as reproduction, low food availability, pollution and parasitism. Birds are affected by diverse parasitic fauna, such as hemoparasites, which may compromise survival and host fitness. The objectives of this work were to examine the leukocyte profile and hemoparasites of a population of Antilophia galeata, and to verify if there is association of both with biological and ecological characteristics of the species. In a Cerrado forest fragment, 151 individuals were captured in 13 field campaigns (06/2013 to 12/2016). The following data were collected: sex, age, biomass, tarsal length, presence/absence of ticks, molt feathers, and brood patch, and blood samples for confection of blood smears, which were analyzed under optical microscope for identification and quantification of leukocytes, micronuclei and parasites. The data collected were divided into two chapters, the first focused on the leukocyte profile, using 89 individuals (captured in 2013-2015) and the second, with a focus on hemoparasites, with 62 individuals (2016). In relation to the leukocyte profile, the number of lymphocytes was higher in males, in non-reproductive phase and in individuals with ticks. There was no difference in any of the leukocyte components in relation to age groups, molt feathers and body condition. The amount of micronuclei was positively related to total leukocyte, H/L ratio, heterophils, basophils and monocytes values. Regarding hemoparasites, 33 (53.2%) individuals were infected, 32 (51.6%) by Plasmodium spp. and one (1.61%) by Haemoproteus spp. There was no difference in parasitemia between the seasons, sexes, presence and absence of reproduction, molt feathers and ectoparasites. There was no correlation between body condition and parasitemia. There was a positive correlation between parasitemia and the total amount of leukocytes, suggesting the increase according to parasitemia, indicating that individuals may be efficient in controlling infections. The present study showed that the leukocyte profile is useful in answering ecological questions, which can be applied in the conservation of the species, because the leukocyte variations are related to stressors such as reproduction and the amount of micronuclei. As for hemoparasites, the results suggest that the population is tolerant to the parasitemia found, since no negative effect was detected. Dissertação (Mestrado)

Purpose Acute lymphoblastic leukemia (ALL) is the most common type of cancer in children, and despite the high rate of cure (over 80%) it still has a big impact on morbidity and mortality. The Transducin-like enhancer of split 1 (TLE1), a transcriptional corepressor, has been described as dysregulated and recently emerged as a tumor marker in several cancer types, including hematologic malignancies. Methods In the present study TLE1 gene expression was evaluated by RT-qPCR. A total of 60 consecutive pathological ALL samples and 8 normal bone marrow samples were included. Associations between TLE1 levels and clinicopathological features were estimated using Mann-Whitney tests. Results TLE1 mRNA levels were significantly diminished in ALL samples when compared to normal counterparts (fold change -1.45, p-value 0.039). Lower TLE1 expression levels were associated with poorer prognostic features such as age at diagnosis ( 9 years-old), absence of the Common Acute Lymphoblastic Leukemia Antigen (CALLA) and high white cell count. Considering immunophenotype, decreased expression of TLE1 was only evident for T-cell ALL, what was validated using gene expression profiling data available in public repositories. No associations with event or overall survival were observed. However, TLE1 expression was statistically different between patients who achieved complete clinical remission (CCR) from those that relapsed or died. Conclusion These data are of particular interest and give support for a plausible role of TLE1 as a tumor suppressor in T-cell ALL. Moreover, the prognostic value of this corepressor may assist ALL treatment stratification and suggest the need of alternative regimens.

Resumo: A indoleamina 2,3-dioxigenase (IDO) é uma enzima que cataboliza o aminoácido triptofano, levando à inibição da proliferação de linfócitos T, seja pela exaustão desse aminoácido no ambiente, ou pela indução via catabólitos induzindo-os a apoptose. Em mamíferos, esta enzima atua em diversas condições do organismo como a gestação, infecções, inflamações crônicas, transplantes e tumores, atuando na regulação imunológica. Estudos recentes identificaram a presença de moléculas homólogas a IDO em espécies filogeneticamente inferiores, cuja função parece estar restrita ao metabolismo do triptofano como fonte de energia. Este estudo teve por objetivo averiguar a expressão da IDO em células sanguíneas e órgãos hematopoiéticos de truta arco-íris pela imuno-histoquímica, buscando evidências de que a mesma poderia, nesta espécie, estar relacionada ao sistema imune. A expressão de IDO foi observada nos órgãos hematopoiéticos estudados incluindo o rim cefálico que apresentou marcação em células interrenais e leucócitos; baço, na qual a marcação restringiu à alguns leucócitos; no fígado a marcação ficou limitada à apenas algumas células dentro dos vasos sanguíneos e nas extensões sanguíneas pode-se visualizar a marcação de alguns leucócitos como os monócitos, linfócitos e neutrófilos. A predominância da marcação da IDO nesses tecidos pode constituir uma evidência de que a IDO identificada na O. mykiss esteja relacionada ao sistema imunológico nessa espécie. Abstract: Indoleamine 2,3-dioxygenase (IDO) is an enzyme that catabolizes the amino acid tryptophan, leading to inhibition of T lymphocyte proliferation, whether by depletion of this amino acid in the environment, or by induction via the catabolites inducing apoptosis. In mammals, this enzyme acts on various conditions of the body such as pregnancy, infections, chronic inflammation, transplantation and tumors, acting in immune regulation. Recent studies have identified the presence of homologous molecules IDO lower phylogenetically related species, whose function appears to be confined to the tryptophan metabolism as an energy source. This study aimed to investigate the expression of IDO in blood cells and hematopoietic organs of rainbow trout by immunohistochemistry, seeking evidence that it could, this species is related to the immune system. The expression of IDO was observed in hematopoietic organs studied including head kidney that show labeling in interrenal cells and leukocytes; spleen, in which the marking restricted to a few leukocytes in the liver;, labeling was restricted to only certain cells within the blood vessels and the blood extensions can view the marking of some leukocytes including monocytes, lymphocytes and neutrophils. The predominance of IDO marking these tissues may constitute evidence that IDO identified in O. mykiss is related to the immune system in this species.

A utilização de bactérias do ácido láctico (LAB) como uma alternativa ao uso de antimicrobianos tem sido proposta para o controle da mastite bovina. No entanto, antes dessas aplicações, o efeito in vivo das LAB sobre a glândula deve ser cuidadosamente avaliado. O objetivo deste estudo foi determinar se a infusão intramamária de Weissella confusa e seus metabólitos em vacas afeta a contagem de células somáticas (CCS) e a contagem diferencial de leucócitos no leite. Vinte e quatro tetos de seis vacas Hartón del Valle foram selecionados para a pesquisa. Uma alíquota de 5mL de uma solução aquosa de células de W. confusa (W) (concentração 109u.f.c. mL-1), 5mL de células de W. confusa com seus metabólitos (W + W10b) ou 5mL de metabólitos (W10b) foi aplicada ao acaso em três tetos de cada vaca. O teto restante em cada úbere representou a unidade experimental de controle (C). Nos dias 1, 2, 3, 5, 7, 9, 13 e 15 após infusão (PI), a CCS, o pH, a acidez titulável e a contagem diferencial de leucócitos no leite foram avaliadas. A aplicação das três substâncias biológicas produziu aumentos significativos na CCS, no pH e na contagem de neutrófilos polimorfonucleares. Não foram encontradas diferenças significativas entre W e W+W10b. O W10b mostrou a menor alteração sobre as variáveis avaliadas. Os resultados sugerem que a aplicação de W. confusa e seus metabólitos afeta a CCS e a contagem diferencial de leucócitos no leite. The use of lactic acid bacteria (LAB) as an alternative to antimicrobial usage has been proposed for the control of bovine mastitis. However, before its application, the in vivo effects of the LAB on mammary gland must be carefully evaluated. The objective of this study was to determine whether the intramammary infusion of Weissella confusa and its metabolites in cows affects the somatic cell counts (SCCs) and milk differential leukocyte count. Twenty-four mammary quarters of six Hartón del Valle cows were selected for the investigation. A 5mL aliquot of an aqueous solution of W. confusa cells (W) (concentration of 109cfu mL-1), 5mL of W. confusa cells with its metabolites (W+W10b) or 5mL of metabolites (W10b) was randomly applied to three mammary quarters of each cow. The remaining teat on each udder represented the experimental control units (C). At 1, 2, 3, 5, 7, 9, 13 and 15 days post-infusion (PI), the SCCs, pH, titrable acidity and milk differential leukocyte count were evaluated. The application of the three biological substances produced significant increases in SCCs, pH and polymorphonuclear neutrophil counts. No significant differences between W and W+W10b infusion were found. W10b resulted in a lesser alteration on the variables evaluated. The results suggest that the intramammary infusion of W. confusa and its metabolites affect the SCCs and milk differential leukocyte count in cows.

Anfíbios são indicadores ambientais potencialmente confiáveis e eficientes. Estudos referentes a morfologia de leucócitos de anuros são limitados, com poucos estudos morfometricos disponíveis em literatura. O presente estudo empregou técnicas morfometricas para caracterizar leucócitos de anuros Neotropicais brasileiros selecionados e compara-los com a espécie exótica rã-touro (Lithobates catesbeianus), família Ranidae. Esfregaços sanguíneos de 28 espécimes pertencentes a seis gêneros diferentes (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus e Proceratophrys) foram comparados com amostras de esfregacos de L. catesbeianus. A média do diâmetro dos leucócitos foi calculada por um software de análise de imagens. One-way e teste de Bonferroni foram utilizados para avaliação estatística. Linfócitos, neutrófilos, eosinófilos e basófilos mostraram-se significativamente menores que os valores de referência reportados em outros gêneros de anfíbios, incluindo Lithobathes; por outro lado, a média do diâmetro dos monócitos não demonstrou variação significativa entre os gêneros. Esse e o primeiro estudo de avaliação morfometrica de leucócitos em espécies de anuros brasileiros. Nossos resultados sugerem que a separação geográfica possivelmente influencia a morfometria leucocitaria. Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry.

Hematologic analysis provides reliable information on the health status of animals. It is an important variable in the assessment of adaptive and productive capacity of breeds under unfavorable environmental conditions. It is an assessment that combined with other genetic and environmental factors and management can become useful for the future sustainability of mainstream agriculture in a hot environment. Adaptive capacity is affected by some factors such as breed, pregnancy and lactation. Therefore, hematological profile is an important indicator of animal health and production. The objective of this study was to perform the comparative hematological analysis of Santa Inês and Morada Nova breeds (native ewes from Brazil) in all different reproductive stages. Twenty Santa Ines and 20 Morada Nova sheep distributed in a completely randomized design in a split-plot arrangement over time were used. To obtain blood counts, blood samples were collected by jugular venipuncture every 14 days, always in the morning, before the animals were released into the pasture. The Morada Nova breed had higher hemoglobin and total leukocyte count than Santa Inês breed. Regarding the influence of pregnancy and puerperium on the eritrogram, there was an increment in red blood cell, hemoglobin and packed cell volume of both breeds in middle pregnancy. However, had a reduction in late pregnancy. There were a reestablishment of the blood cell counts during the puerperium period. It was verified an increase on neutrophil: lymphocyte ratio indicated that ewes in the late pregnancy, postpartum and puerperium were under stress conditions compared to others reproductive stages. The total plasma proteins also increased during this period to compensate for the high nutritional requirements of the fetus, and these levels remained high until the end of the puerperal phase, when the lambs were weaned. The pregnancy and puerperium influenced all erythrocyte indices and changed the total leukocyte count. RESUMO: A análise hematológica fornece informação confiável sobre a saúde dos animais. É uma variável importante na avaliação da capacidade adaptativa e produtiva das raças sob condições ambientais desfavoráveis. É uma avaliação que aliada a outros fatores genéticos, ambientais e de manejo pode se tornar cada vez mais útil para a sustentabilidade futura da agricultura convencional em um ambiente quente. A capacidade adaptativa é afetada por alguns fatores como raça, gestação e lactação. Portanto, o perfil hematológico é um indicador importante da saúde e produção animal. O objetivo deste estudo foi realizar a análise hematológica comparativa de ovelhas Morada Nova e Santa Inês (ovelhas nativas do Brasil) em todos os estágios reprodutivos. Foram utilizadas 20 ovelhas Santa Inês e 20 Morada Nova distribuídas em um delineamento inteiramente casualizado, em parcelas subdivididas e medidas repetidas no tempo. Para a análise do sangue, foram coletadas amostras por venopunção jugular a cada 14 dias, sempre colhidas pela manhã, antes que os animais fossem soltos no pasto. A raça Morada Nova apresentou maior teor de hemoglobina e contagem total de leucócitos que a raça Santa Inês. Em relação à influência da gestação e do puerpério sobre o eritrograma, observou-se que houve um incremento dos valores da contagem dos eritrócitos, hematócrito e hemoglobina até o terço médio da gestação, reduzindo no final da gestação e restabelecendo novamente no período puerperal. Foi verificado um aumento na relação neutrófilo/linfócito indicando que ovelhas no final da gestação, no pós-parto e puerpério estavam sob condição de estresse comparado com os outros estágios reprodutivos. As proteínas plasmáticas totais também aumentaram durante o período gestacional para compensar os altos requisitos nutricionais do feto, e estes níveis permaneceram elevados até ao final da fase de puerpério quando os cordeiros foram desmamados. A gestação e o puerpério influenciaram todos os índices eritrocitários com alteração da contagem total de leucócitos.

RESUMO: A avaliação hematológica, de importância comprovada como um meio auxiliar de diagnóstico ao clínico de pequenos animais domésticos, vem se tornando comum em animais selvagens não apenas para a clínica, mas para a avaliação do manejo e como estudo auxiliar para a fisiologia das várias espécies. Tendo em vista o aumento da demanda para a produção de várias drogas de importância farmacêutica, a criação de serpentes peçonhentas vem se tornando comum a ponto destes animais já serem reconhecidos como sendo de produção. O conhecimento do manejo e da clínica destes animais ainda é escasso e a mortalidade é elevada nos criatórios, tornando urgente a ampliação destes. Embora alguns estudos hematológicos já tenham sido realizados em cascavéis (Crotalus durissus) os dados analisados ainda são insipientes, notadamente em relação à caracterização das células do sangue e poucos estudos em microscopia eletrônica foram realizados em serpentes. Com o objetivo de caracterizar as células sanguíneas morfologicamente, sob microscopia óptica e ultraestrutural, foram coletadas amostras de sangue de 52 de indivíduos da subespécie Crotalus durissus terrificus para a realização de esfregaços sanguíneos e avaliação ultraestrutural. Concluiu-se que a coloração hematológica de Giemsa permite a avaliação morfológica e a diferenciação das células sanguíneas em serpentes assim como a visualização de hemoparasitos. A avaliação ultraestrutural permite evidenciar as organelas celulares e a diferenciação entre as células, inclusive entre os tipos leucocitários, porém ainda são necessários outros estudos para que seja elucidada a hipótese da existência dos eosinófilos na espécie estudada assim como é necessária melhor caracterização dos grânulos dos azurófilos para que se confirme uma possível diferença entre os monócitos típicos e os azurófilos. ABSTRACT: Hematological evaluation, important for the diagnostic by the small domestic animal clinician, has become common in wildlife clinic, and for handling and study of the physiology of various species. Given the increased demand for drug production of pharmaceutical importance, the breeding of venomous snakes has become common and is already recognized as production. Knowledge of the management and clinics of snakes is still insufficient and their mortality is high. Although some hematological studies have already been conducted in the rattlesnake (Crotalus durissus), the analyzed data are still insufficient, especially with respect to the characterization of blood cells, and few electron microscopy studies have been performed on snakes. In order to characterize morphologically blood cells with light and ultrastructural microscopy, blood samples from 52 individuals of subspecies of Crotalus durissus terrificus were collected to perform blood smears and ultrastructural evaluation. It was concluded that hematologic Giemsa staining allows morphological evaluation and differentiation of the blood cells as well as of snake hemoparasites. The ultrastructural evaluation will highlight the cell organelles and differentiation between cells, including leukocyte types; although still further studies are needed to elucidate the hypothesis of eosinophils in the species studied as also is necessary a better characterization of azurophilic beads to confirm a possible difference between the typical monocyte and the azurophilic.

Reptiles are the unique ectothermic amniotes, providing the key link between ectothermic anamniotes fish and amphibians, and endothermic birds and mammals; becoming an important group to study with the aim of providing significant knowledge into the evolutionary history of vertebrate immunity. Classification systems for reptiles' leukocytes have been described by their appearance rather than function, being still inconsistent. With the advent of modern techniques and the establishment of analytical protocols for snakes' blood by flow cytometry, we bring a qualitative and quantitative assessment of innate activities presented by snakes' peripheral blood leukocytes, thereby linking flow cytometric features with fluorescent and light microscopy images. Moreover, since corticosterone is an important immunomodulator in reptiles, hormone levels of all blood samples were measured. We provide novel and additional information which should contribute to better understanding of the development of the immune system of reptiles and vertebrates.

Se recolectaron 69 muestras de Trachemys callirostris en condiciones ex situ e in situ, para determinar sus parámetros hematológicos, comparar estos valores (considerando sexo, edad y condición) y establecer la morfología de sus células. A los animales se les extrajo sangre de la vena yugular. Se determinaron hematocrito, concentración de hemoglobina, recuentos total de eritrocitos, leucocitos y trombocitos e índices eritrocitarios (volumen corpuscular medio, hemoglobina corpuscular media y concentración de hemoglobina corpuscular media). En extendidos sanguíneos se realizaron el conteo diferencial de leucocitos y la descripción de la morfología de las células encontradas. Los valores promedio encontrados para todos los individuos fueron: hematocrito (29,9 % ± 5,1), hemoglobina (4,0 ± 1,0 g/dl), VCM (327 ± 15,3 fl), HCM (45 ± 17,5 pg), CMHC (14 ± 6 %), conteo total de trombocitos (15,0 ± 5,4 × 10³ μL), conteo total de eritrocitos (0,92 × 10⁶/μL ± 0,12), conteo total de leucocitos (12,10 ± 5,0 × 10³ μL). Al aplicar el análisis de varianza se encontró que no existe diferencia estadísticamente significativa (p < 0,05) entre Trachemys callirostris de distintos pesos (500 a 2000 g). Sin embargo, existen diferencias significativas (p < 0,05) y altamente significativas (p < 0,01) de la mayoría de las variables hematológicas entre sexos y entre hicoteas mantenidas en condición in situ y ex situ. De esta forma, las diferencias encontradas entre individuos, dependiendo su peso, sexo y condición de mantenimiento (in situ o ex situ), sirven como referencia para el establecimiento de protocolos de manejo de la especie en cautiverio.

Recruitment of a specific cell population after Leishmania infection can influence the outcome of the disease. Cellular migration in response to Leishmania or vector saliva has been reported in air pouch model, however, cellular migration induced by Leishmania associated with host's blood and vector saliva in this model has not been described. Herein we investigated cellular migration into air pouch of hamster after stimulation with combination of L. chagasi and host's blood and Lutzomyia longipalpis saliva. Migration induced by saliva was 3-fold more than those induced by L. chagasi alone. Additionally, L. chagasi associated with blood and saliva induced significantly even more leukocytes into air pouch than Leishmania alone. L. chagasi recruited a diverse cell population; however, most of these cells seem to have not migrated to the inflammatory exudate, remaining in the pouch lining tissue. These results indicate that L. chagasi can reduce leukocyte accumulation to the initial site of infection, and when associated with vector saliva in the presence of blood components, increase the influx of more neutrophils than macrophages, suggesting that the parasite has developed a strategy to minimize the initial inflammatory response, allowing an unlimited progression within the host. This work reinforces the importance of studies on the salivary components of sand fly vectors of leishmaniasis in the transmission process and the establishment of the infection.

There is a growing demand for alternative protein sources in dog nutrition due to the high cost of the ingredient. Spray-dried egg (SDE) is a potential protein source in this context. The objective of this study was to evaluate the digestibility, palatability, and blood profile of diets containing different amounts of spray-dried egg (SDE) and the blood parameters of dogs fed these diets. For the study of digestibility and faecal quality (Experiment 1), eight Beagles were distributed in a double Latin square design (4X4), with four treatments (0, 4, 8, and 12% SDE) and four periods, totalling eight repetitions per treatment. In experiment 2, to evaluate palatability, 20 dogs were presented with two diets: 0% SDE vs 4% SDE. To evaluate the haematological factors (Experiment 3), eight dogs were maintained in a single group and were initially fed a control diet (0%SDE) for 10 days, followed by a diet containing 12% SDE for 10 days. In Experiment 1, a linear increase was found in the coefficients of total tract apparent digestibility (CTTAD) of dry matter (DM) and crude protein (CP) and in the metabolisable energy (ME) as the level of SDE was increased in the diet. The CTTAD of DM and CP in SDE and the ME obtained by regression analysis were 93.6%, 91.3%, and 5738.9 kcal kg-1, respectively, demonstrating that SDE is a highly digestible protein source for dogs. There were no changes in the stool characteristics of dogs fed SDE. The results of Experiment 2 indicate that the dogs preferred the diet that included 4% SDE. In the haematological study (Experiment 3), increases in albumin, red blood cells, haemoglobin, and total circulating protein were observed, with no change in the other histological variables. The inclusion of SDE improved the digestibility and palatability of the diet and increased the albumin content, red blood cell count, haemoglobin level, and total protein level without changing the faecal characteristics. Há uma demanda crescente de fontes proteicas alternativas na nutrição de cães devido ao alto custo deste ingrediente. O ovo seco por pulverização (SDE) é uma fonte potencial de proteína neste contexto. O objetivo deste estudo foi avaliar a digestibilidade, palatabilidade e perfil sanguíneo de dietas contendo diferentes níveis de ovo spray-dried (OSD) e os parâmetros sanguíneos de cães alimentados com esta dieta. Para o estudo de digestibilidade e características fecais (Experimento 1), oito cães foram distribuídos em delineamento quadrado latino duplo (4x4), com quatro tratamentos (0, 4, 8 e 12% OSD) e quatro períodos, totalizando oito repetições por tratamento. No experimento 2, para avaliar a palatabilidade, 20 animais foram apresentados a duas dietas: 0% OSD vs 4% OSD. Para avaliar os fatores hematológicos (Experimento 3), oito cães foram mantidos em um único grupo e foram inicialmente alimentados com a dieta controle (0% OSD) por 10 dias, seguido pela dieta contendo 12% OSD por 10 dias. No experimento 1, foi observado aumento linear para os coeficientes de digestibilidade aparente total (CDAT), da matéria seca (MS), e da proteína bruta (PB) e a energia metabolizável (EM) com o aumento de OSD da dieta. Os CDA da MS e PB em OSD e a EM obtidas pela análise da regressão foram de 93.6%, 91.3% e 5738.9 kcal kg-1, respectivamente, demonstrando que o OSD é uma fonte proteica de alta digestibilidade para cães. Não houve mudanças nas características fecais de cães alimentados com OSD. Os resultados do experimento 2 indicam que os cães preferiram a dieta com inclusão de 4% de OSD. No estudo hematológico (Experimento 3), aumento de albumina, células vermelhas do sangue, hemoglobina e proteína total circulante foram observados, não houve mudança em outras variáveis histológicas. A inclusão de OSD melhorou a digestibilidade e palatabilidade da dieta e aumentou o teor de albumina, a contagem de células vermelhas, o nível de hemoglobina e o nível de proteína total sem alterar as características fecais.

INTRODUÇÃO: O triatlo Ironman se caracteriza por ser uma atividade de longa duração em que alterações orgânicas agudas estão presentes. OBJETIVO: verificar a ocorrência de dano muscular e sua relação com o perfil imunológico em triatletas do Ironman - Brasil. MÉTODOS: A amostra de sangue foi obtida de 21 atletas em três momentos: dois dias antes da prova (pré), imediatamente após a prova (pós) e seis dias após a prova (seis dias pós), em que foram analisadas de forma isolada as variáveis creatinoquinase (CK), os leucócitos totais, linfócitos, subtipos de linfócitos CD4+ e CD8+, e relação CD4+/CD8+ e a correlação da CK como marcador de dano muscular, com as demais variáveis. RESULTADOS: As diferenças significativas foram observadas nos leucócitos pré (média: 6.242,9 mm³; DP: 1.233,3) e pós (média: 18.398,1 mm³; DP: 3.904,0; p < 0,0001); pós (média: 18.398,1 mm³; DP: 3.904,0) e seis dias pós (média: 6.396,4 mm³; DP: 1.299,8; p < 0,0001); CK pré (média: 173,2 U/l; DP: 103,7) e pós (média: 2.339,4 U/l; DP: 1.729,0; p < 0,0001), CK pré (média: 173,2 U/l; DP: 103,7) e seis dias pós (média: 368,1 U/l; DP: 274,4; p < 0,0053); CK pós (média: 2.339,4 U/l; DP: 1.729,0) e seis dias pós (média: 368,1 U/l; DP: 274,4; p < 0,0003); CD4+/CD8+ pré (média: 1,9; DP: 0,8) e seis dias pós (média: 2,4; DP: 1,1: p < 0,00032). CONCLUSÃO: Houve dano muscular no período pós-prova imediato e melhora do perfil imunológico após o sexto dia.

Critically short telomeres activate p53-mediated apoptosis, resulting in organ failure and leading to malignant transformation. Mutations in genes responsible for telomere maintenance are linked to a number of human diseases. We derived induced pluripotent stem cells (iPSCs) from 4 patients with aplastic anemia or hypocellular bone marrow carrying heterozygous mutations in the telomerase reverse transcriptase (TERT) or the telomerase RNA component (TERC) telomerase genes. Both mutant and control iPSCs upregulated TERT and TERC expression compared with parental fibroblasts, but mutant iPSCs elongated telomeres at a lower rate compared with healthy iPSCs, and the deficit correlated with the mutations' impact on telomerase activity. There was no evidence for alternative lengthening of telomere (ALT) pathway activation. Elongation varied among iPSC clones derived from the same patient and among clones from siblings harboring identical mutations. Clonal heterogeneity was linked to genetic and environmental factors, but was not influenced by residual expression of reprogramming transgenes. Hypoxia increased telomere extension in both mutant and normal iPSCs. Additionally, telomerase-mutant iPSCs showed defective hematopoietic differentiation in vitro, mirroring the clinical phenotype observed in patients and demonstrating that human telomere diseases can be modeled utilizing iPSCs. Our data support the necessity of studying multiple clones when using iPSCs to model disease.

Inflammation plays a key role in the development of insulin resistance and atherosclerosis. Fatty acids and fiber intake can selectively alter gene expression by modifying inflammation. We compared the postprandial expression of inflammatory genes after 2 distinct high-fat breakfast meals, before and after 1-month dietary interventions. This crossover clinical trial included 18 individuals at low-to-moderate cardiometabolic risk participating in evaluations before and after two 4-week breakfast interventions—one rich in saturated fatty acids (SFA) and the other in unsaturated fatty acids (unSFA) and fiber. Participants underwent meal tests with similar compositions to the breakfasts. Variables were compared by Student t test. The expression of inflammatory genes in leukocytes was analyzed using RT-PCR. Before and after the intervention with the SFA-enriched breakfast, this meal test induced a higher relative postprandial IL-1β expression compared to the responses to the unSFA and fiber-enriched meal (p = 0.02). On the other hand, following the intervention with the unSFA-fiber-enriched breakfast, postprandial IL-6 expression showed a reduction tendency comparing to the pre-intervention value (p = 0.08). Although fasting IL-1β, IL-6, MCP-1 and IFN-γ expressions had not changed after interventions, their circulating levels increased after the SFA-enriched meal test but not after the unSFA meal (p value between changes

Resumo: Peixes-boi da Amazônia (Trichechus inungui) jovens são frequentemente encontrados e resgatados de rios, no entanto para avaliar e monitorar o estado de saúde desses animais é necessário conhecer os valores de referência das variáveis hematológicas e bioquímicas nessa faixa etária. O trabalho objetivou determinar os valores hematológicos e bioquímicos de peixes-boi da amazônia jovens e saudáveis, mantidos em cativeiros. Foram coletadas 20 amostras sanguíneas de peixes-boi para a realização do hemograma e bioquímica. Os valores médios e desvio padrão das variáveis hematológicas foram: volume globular, 37±4%; eritrócitos, 4,4±0,35x106/μL; hemoglobina, 12,5±1,6g/dL; volume corpuscular médio (VCM), 85,33±4,28fL; hemoglobina corpuscular media (HCM), 28,29±1,39pg; e concentração de hemoglobina corpuscular media (CHCM), 33,13±0,20%, leucócitos totais, 9.080±1.868x103/μL; segmentados, 5.078±1.044,24x103/μL; linfócitos 3.556±1,063x103/μL; eosinófilos, 288± 215,51x103/μL; e monócitos, 165± 114,11x103/μL. A média e o intervalo (mínimo e máximo) das variáveis bioquímicas foram: creatinina quinase (CK), 112,53 (50,10-295,80) U/L; aspartato aminotransferase (AST), 6,02 (2,80-10,20) U/L; gama glutamiltransferase (GGT), 25,24 (10,80-45,30) U/L; fosfatase alcalina (FA), 98,73 (14,66-198,30) U/L; proteína total, 6,12 (5,38-7,10) g/dL; albumina, 4,23 (3,50-5,00) g/dL; colesterol, 310,13 (144-518) mg/dL; triglicerídeos, 127,98 (57,50-213,50) mg/dL; glicose, 43,63 (24,50-73,80) mg/dL; ureia, 30,21 (14,30-54,25); creatinina, 1,34 (0,93-1,76) mg/dL; cálcio, 10,08 (9,25-11,10) mg/dL; e fósforo inorgânico; 6,50 (4,10-8,80) mg/dL. No presente trabalho foi possível determinar valores hematológicos e bioquímicos de peixes-boi da Amazônia (Trichechus inungui) jovens, mantidos em cativeiro, os quais podem ser utilizados como referência para animais em cativeiro e de vida livre, mantidos em condições ambientais, e de manejo, semelhantes. Abstract: Amazonian manatee (Trichechus inunguis) young are often found and rescued from rivers, however to assess and monitor the health status of the animals is necessary to know the reference values of hematological and biochemical variables in this age group. The study aimed to determine hematological and biochemical values of healthy young manatee amazonian, kept in captivity. Were collected 20 blood samples of manatees in captivity for the hematological and biochemical evaluation. Means and standard deviations of hematological variables were: pack cell volume, 37±4%; red blood cell, 4.4±0.35x106/μL; hemoglobin, 12.5±1.6g/dL; mean corpuscular volume (VCM), 85.33±4.28fL; mean corpuscular hemoglobin (HCM), 28.29±1.39pg; and mean corpuscular hemoglobin concentration (CHCM), 33.13±0.20%. White blood cell, 9,080±1,868x103/μL; neutrofil, 5,078±1,044.24x103/μL; lymphocytes 3,556±1,063x103/μL; eosinophil, 288±215.51x103/μL; and monocyte, 1.65±114.11x103/μL. Means and ranges (minimum and maximum) of values for the analytes were: creatine kinase (CK), 112.53 (50.10-295.80) U/L; aspartate aminotransferase (AST), 6.02 (2.80-10.20) U/L; γ-glutamyltransferase (GGT), 25.24 (10.80-45.30) U/L; alkaline phosphatase (FA), 98.73 (14.66-198.30) U/L; total protein, 6.12 (5.38-7.10) g/dL; albumin, 4.23 (3.50-5.00) g/dL; cholesterol, 310.13 (144-518) mg/dL; triglycerides, 127.98 (57.50-213.50) mg/dL; glucose, 43.63 (24.50-73.80) mg/dL; urea, 30.21 (14.30-54.25); creatinine, 1.34 (0.93-1,76) mg/dL; calcium, 10.08 (9.25-11.10) mg/dL; and inorganic phosphorum, 6.50 (4.10-8.80) mg/dL. In the present study we determined hematological and biochemical values of manatees Amazonian (Trichechus inunguis) young held in captivity, which can be used as a reference for animals in bondage and free life, kept under similar ambient conditions, and management similar.

Background: Identification of asthma phenotypes enables a better understanding and management of this heterogeneous disease. Studies have reported associations between human leukocyte antigens (HLA) and asthma in different populations, but the results have been inconclusive and they have rarely considered the distinct disease phenotypes. Our objective was to characterize allergic and nonallergic asthma phenotypes and evaluate possible associations with the HLA system. Methods: A total of 109 patients with asthma were prospectively followed during 2 years. They were divided into 2 groups, i.e., allergic and nonallergic asthma, according to their clinical history and skin prick test and serum-specific immunoglobulin E (IgE) results. The control group comprised 297 deceased donors of solid organs. Patients' features and HLA class I and II genotypes were assessed and compared. Results: This study showed different features between asthma phenotypes. Nonallergic patients were older at the onset of asthma symptoms and had a higher rate of intolerance to nonsteroidal anti-inflammatory drugs. Allergic patients had higher total serum IgE levels, reported atopic dermatitis and rhinoconjunctivitis more frequently, and, unexpectedly, had a greater disease severity. New associations between the HLA genotypes and allergic and nonallergic asthma were identified. The HLA-B*42, HLA-C*17, HLA-DPA1*03, and HLA-DPB1*105 genotypes were associated with allergic asthma and the HLA-B*48 genotype with the nonallergic phenotype. The presence of the haplotype HLA-DPA1*03 DQA*05 was associated with allergic asthma, and the presence of HLA-DPA1*03 and the absence of HLA-DQA*05 with nonallergic asthma. Conclusions: Allergic and nonallergic asthma have distinct phenotypic and genotypic features. New associations between asthma phenotypes and HLA class I and II were identified.

Background Physiologically, a reduction in telomere length (LTL) occurs with aging, but epigenetic changes may accelerate telomere shortening and also facilitate the onset of oxidative/inflammatory stress and the development of clinical/metabolic comorbidities in life spam. Although individuals born small for gestational age (SGA) may be related to those epigenetic changes, the assessment of LTL in individuals born SGA has yielded conflicting results (only cross-sectional studies) and has not been carried out in longitudinal studies. We performed a birth cohort study to evaluate the rate of telomere erosion in women born SGA in comparison to women born appropriate for gestational age (AGA) assessed at two different time points during the third decade of life. In our research, born SGA or AGA showed no difference in LTL shortening during a period of five years in the third decade of life. Our finding may have implications for understanding the natural history of diseases in lifespan because the same women (under the influence of similar environmental factors) may be accessed in different phases of life. Thus, the analysis of the present cohort population at a more advanced age may reveal a dynamics of telomere shortening different from here and its possible relation with onset of age-related diseases.

Background: Eicosanoids mediate initiation and resolution of inflammation. Our aim was evaluating the effects of training, exercise and docosahexaenoic (DHA) supplementation on plasma eicosanoids levels and peripheral blood mononuclear cells (PBMCs) eicosanoids production. Methods: Fifteen male footballers were distributed to placebo and experimental groups. Experimental group consumed DHA-enriched beverage (1.16 g DHA/day) for 8 weeks, placebo group consumed a placebo beverage. Blood samples were taken before and after the nutritional intervention in basal conditions and 2 h after acute exercise. Results: Training increased basal Prostaglandin E1 (PGE(1)) plasma levels and PBMCs cyclooxygenase 2 (COX-2) protein levels in both groups, but COX-1 protein levels only in the experimental group. Acute exercise increased plasma PGE(2) and PBMCs active NF kappa beta levels. Lipopolysaccharide (LPS)-stimulated PBMCs increases eicosanoids production (PGE(1), PGE(2), RvD1) in both groups and increased LPS-stimulated PBMCs active NF kappa beta. DHA supplementation increased COX-2 levels but decreased LPS-stimulated PBMCs PGE(1) and PGE(2) production. Neither DHA supplementation nor acute exercise altered the expression of NF kappa beta, COX-2, 15-LOX2, 5-LOX, or IL-1 beta genes in PBMCs. Conclusions: The increase of PGE(1) plasma levels after training promoted systemic anti-inflammatory and vasodilator environment. Exercise and DHA supplementation acted synergistically by increasing plasma PGE(2) with anti-inflammatory effects. Exercise primed PBMCs to enhance PGE(1), PGE(2) and RvD1 production in response to LPS., Accion Estrategica en Salud del Ministro de Ciencia e Innovacion DPS2008-07033-C03-03, Programme of Promotion of Biomedical Research and Health Sciences, Projects 11/01791, Red Predimed-RETIC RD06/0045/1004, CIBERobn CB12/03/30038 and Balearic Island Government and FEDER founds (35/2011 and 23/2012). We hereby acknowledge the PhD grant provided by the University of the Balearic Islands.

The weaning period of piglets is characterized by physiological alterations, such as decreased weight gain, increased reactive oxygen species (ROS) and increased serum cortisol levels with possible effects on the immune response. The effect of parenteral administration of vitamins A, D and E on production performance, oxidative metabolism, and the function of polymorphonuclear leukocytes (PMNLs) was assessed in piglets during the weaning period. The sample was comprised of 20 male piglets that were given an injectable ADE vitamin combination (135,000 IU vitamin A, 40,000 IU vitamin D and 40mg vitamin E/ animal) at 20 and 40 days of age. Weight gain, concentration of reduced glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD) and the microbicidal and phagocytic activity of PMNLs were assessed. No difference was observed in the average piglet weight during the study; however, a greater percentage of weight gain was observed after weaning in the treated group. The concentrations of GSH and SOD did not differ between groups, although lipid peroxidation was greater in the control group at 60 days of age. The investigated variables of oxidative metabolism were correlated as follows: -0.41 for GSH and MDA, -0.54 for GSH and SOD and 0.34 for MDA and SOD. The intensity of intracellular ROS production, the percentage of ROS-producing PMNLs and the intensity of phagocytosis by PMNLs did not differ between treatment groups. Administration of the injectable ADE combination improved the percentage of weight gain between 20 and 40 days of age, decreased oxidative stress at 60 days of age and did not influence the function of PMNLs in piglets. O período de desmame nos leitões é caracterizado por alterações fisiológicas como menor ganho de peso, aumento na produção de espécies reativas de oxigênio (EROs) e aumento na concentração plasmática de cortisol com possíveis implicações para a resposta imune. Foi avaliado o efeito da administração parenteral das vitaminas A, D e E sobre o desempenho produtivo, o metabolismo oxidativo e a função de leucócitos polimorfonucleares (PMNLs) em suínos durante esta fase de crescimento. Foram utilizados 20 leitões, machos, com 20 dias de idade que receberam ADE injetável (135.000 UI vitamina A, 40.000 UI vitamina D e 40mg vitamina E/animal), aos 20 e 40 dias de idade. Foi determinado o ganho de peso e as concentrações de glutationa reduzida (GSH), malondialdeído (MDA) e superóxido dismutase (SOD) e a capacidade microbicida e fagocítica dos PMNLs. Não houve diferença entre o peso vivo médio durante o experimento, porém maior ganho de peso percentual foi observado 20 dias após o desmame para o grupo tratado. As concentrações de GSH e SOD não diferiram entre os grupos, porém a lipoperoxidação foi maior no grupo controle aos 60 dias de idade. As correlações entre as variáveis do metabolismo oxidativo foram -0,41 para GSH e o MDA, -0,54 para GSH e SOD e 0,34 para MDA e SOD. A intensidade da produção intracelular de EROs, a porcentagem de PMNLs que produziram EROs e a intensidade de fagocitose dos PMNLs não diferiram entre os tratamentos. A administração de ADE injetável melhorou o ganho de peso percentual no período de 20 a 40 dias de idade, diminuiu o estresse oxidativo aos 60 dias de idade e não influenciou função dos PMNLs dos leitões.

Telomeres, the natural ends of linear chromosomes, cap and protect chromosomes against damage and from being mistaken for double-stranded DNA breaks.(1) When a cell divides, telomeres shorten due to DNA polymerase’s inability to fully replicate the 3′ ends of chromosomes, the “end-under-replication problem”.(2) If telomeres become critically short, cellular signaling cascades involving p53 and p21 are activated, resulting in cell senescence or apoptosis.(3, 4) To counter telomere attrition, cells with high proliferative capacity express telomerase, a reverse transcriptase enzyme that adds DNA repeats to telomeres.(5) Components of the telomerase complex include the reverse transcriptase enzyme, TERT, the RNA component that serves as template for telomere elongation, encoded by TERC, and associated proteins, including dyskerin(encoded by DKC1).(6, 7) Loss-of-function heterozygous or homozygous mutations in genes of the telomerase complex have been implicated in human diseases caused by a deficient tissue regeneration capacity,(3)including dyskeratosis congenita(8, 9), aplastic anemia,(10, 11)and familial idiopathic pulmonary fibrosis,(12, 13)and also with acute myeloid leukemia.(14, 15) Common genetic variants in the region of the TERT gene on chromosome 5p15.33 have been associated with susceptibility to braintumors, lung cancer, pancreatic cancer and additional cancers(16–20). Dyskeratosis congenita is a rare genetic disease in which patients develop bone marrow failure and exhibit a mucocutaneous triad of abnormal reticular skin pigmentation, leukoplakia, and nail dystrophy.(21)Most cases are X-linked and caused by mutations in the DKC1 gene. Dyskeratosis congenita also may be autosomal dominant, in which heterozygous mutations in the telomere biology genes TERT, TERC, or TINF2 are etiologic, or autosomal recessive, due to mutations in NOLA2 or NOLA3, coding telomerase-associated proteins.(22, 23) The observation that lungdisease, mainly pulmonary fibrosis, is present in up to 20% of patients with dyskeratosis congenita led to the association of telomerase mutations with familial idiopathic pulmonary fibrosis.(12) Approximately, seven percent of dyskeratosis patients have a concurrent diagnosis of hepatic disease, including cirrhosis.(21) Fatal liver complications are a relatively common cause of death after hematopoietic stem-cell transplantation for bone marrow failure in dyskeratosis congenita, whereas fatal liver complications are infrequent following transplant for other disorders, suggestive of a related underlying mechanism.(24) In families of patients with telomerase mutation and aplastic anemia, severe hepatic disease in relatives tracks to mutation status.(25) The relationship between telomere shortening and risk of cirrhosis has been examined in an experimental model of mice null for the telomerase reverse transcriptase gene. Tert-deficient mice had reduced regenerative activity following partial hepatectomy as well as more hepatic fibrosis and inflammation after exposure to CCl4 compared to normal mice.(26) In human cirrhosis, hepatocytes display excessive telomere shortening and senescence.(27, 28) These findings in experimental animals and observations in humans suggest that reduced telomerase activity may contribute to the development of cirrhosis. In the present study, we sought to determine whether germ-line missense sequence variants in the telomerase complex genes are more frequent in patients with non-familial cirrhosis due to a variety of causes.

A leishmaniose visceral (LV) é uma zoonose causada pelo protozoário Leishmania (Leishmania) chagasi. A leishmaniose visceral canina (LVC) é a doença de maior relevância zoonótica. Usualmente, a infecção ocorre entre um hospedeiro invertebrado para um hospedeiro vertebrado, entretanto, a transmissão na ausência do vetor já é conhecida. O objetivo principal deste estudo foi identificar a presença de formas amastigotas, quantificar as células leucocitárias, estimar o risco relativo da presença de formas amastigotas no aparelho reprodutivo de cães sorologicamente positivos com e sem sinais clínicos. Para isso, foram utilizados cães sem raça definida, sexualmente maduros e testados sorologicamente para LVC (com sinais clínicos, n=25; sem sinais clínicos, n=25), que após eutanásia, tiveram fragmentos de testículo, epidídimo (cabeça, corpo e cauda) e glândula prostática (selecionados ao acaso) impressos em lâminas. Um grupo de 20 cãs sorologicamente negativos e sem sinais clínicos foi usado como controle. Amostras do baço foram incluídas como controle parasitológico positivo. O percentual de linfócitos foi superior (P

Objetivos. Evaluar el efecto antiinflamatorio de la fracción flavonoide de Lepechinia meyenii (Walp.) Epling sobre leucocitos de pacientes con artritis reumatoide (AR). Materiales y métodos. Se recolectaron plantas de la especie Lepechinia meyenii (Walp.) Epling extrayendo diferentes fracciones flavonoides por cromatografía de columna y de capa fina. Se evaluó la producción de anión superóxido mediante la técnica de ensayo reducción nitroblue tetrazolium, en neutrófilos obtenidos de sangre de pacientes con AR, separados en tres grupos: control negativo, que consistió de neutrófilos (5x10(5) células), control positivo, formado por neutrófilos activados con PMA (phorbol myristate acetate) (150 ng/mL) y los tratamientos, formados por neutrófilos activados y tratados con diferentes concentraciones de la fracción flavonoide LM8 (60, 120 y 180 ug/mL). La expresión de genes proinflamatorios se estudió por RTqPCR, en leucocitos mononucleares obtenidos de pacientes con AR separados en tres grupos: control negativo, que consistió de leucocitos mononucleares (5x10(5)células), control positivo formado por leucocitos mononucleares activados con fitohemaglutinina (PHA) (150 ug/mL) y el tratamiento formado por leucocitos mononucleares activados y tratados con la fracción flavonoide LM8 (120 ug/mL). Resultados. Se purificaron varias fracciones flavonoides, resultando la fracción LM8 con el mejor efecto inmunomodulador. Dicha fracción disminuyó la producción de anión superóxido en una manera dependiente de la concentración. Por otro lado, disminuyó la expresión de TNFα, IL8 e IL17 en leucocitos mononucleares Conclusiones. Estos resultados son alentadores respecto al efecto inmunomodulador de esta planta medicinal peruana y justifican continuar su estudio para una posible aplicación clínica. Objectives. to assess the anti-inflammatory effect of the flavonoid fraction of Lepechinia meyenii (Walp.) Epling on leukocytes of patients with rheumatoid arthritis (RA). Materials and Methods. Plants of the species Lepechinia meyenii (Walp.) Epling were collected and then different flavonoid fractions were extracted by column and thin layer chromatography. The superoxide anion production was evaluated by means of the reduction of nitroblue tetrazolium assay technique in neutrophils obtained from the blood of patients with RA, divided into three groups: negative control, which consisted of neutrophils (5x10(5) cells); positive control, made up of PMA (phorbol myristate acetate)-activated neutrophils (150 ng/mL), and the treatments, comprised of neutrophils activated and treated with different concentrations of the flavonoid fraction LM8 (60, 120, and 180 ug/mL). The expression of pro-inflammatory genes was studied by RTqPCR in mononuclear leukocytes obtained from patients with RA, divided into three groups: negative control, which consisted of mononuclear leukocytes (5x10(5) cells); positive control, made up of phytohaemagglutinin (PHA) (150 ug/ml)-activated mononuclear leukocytes, and the treatment, comprised of mononuclear leukocytes activated and treated with the flavonoid fraction LM8 (120 ug/mL). Results. Several flavonoid fractions were purified, with fraction LM8 showing the best immunomodulating effect. Said fraction diminished the superoxide anion production dependent on concentration. On the other hand, it diminished the expression of TNFα, IL8, and IL17 in mononuclear leukocytes. Conclusions. These results are encouraging in terms of the immunomodulating effect of this Peruvian medicinal plant and justify the continuation of their study for a potential clinical application.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:56:02Z No. of bitstreams: 1 S0102-09352008000200007.pdf: 204124 bytes, checksum: 86b02f359a911d19128045a2ff0b3d4f (MD5) Made available in DSpace on 2013-08-22T18:56:02Z (GMT). No. of bitstreams: 1 S0102-09352008000200007.pdf: 204124 bytes, checksum: 86b02f359a911d19128045a2ff0b3d4f (MD5) Previous issue date: 2008-04-01 Made available in DSpace on 2013-09-30T19:49:32Z (GMT). No. of bitstreams: 2 S0102-09352008000200007.pdf: 204124 bytes, checksum: 86b02f359a911d19128045a2ff0b3d4f (MD5) S0102-09352008000200007.pdf.txt: 25205 bytes, checksum: 555d8a6e12f05be68cf720ddf25dc051 (MD5) Previous issue date: 2008-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:14:08Z No. of bitstreams: 2 S0102-09352008000200007.pdf: 204124 bytes, checksum: 86b02f359a911d19128045a2ff0b3d4f (MD5) S0102-09352008000200007.pdf.txt: 25205 bytes, checksum: 555d8a6e12f05be68cf720ddf25dc051 (MD5) Made available in DSpace on 2014-05-20T13:14:08Z (GMT). No. of bitstreams: 2 S0102-09352008000200007.pdf: 204124 bytes, checksum: 86b02f359a911d19128045a2ff0b3d4f (MD5) S0102-09352008000200007.pdf.txt: 25205 bytes, checksum: 555d8a6e12f05be68cf720ddf25dc051 (MD5) Previous issue date: 2008-04-01 Foram examinados 20 eqüinos adultos portadores de abdômen agudo e submetidos à laparotomia. Dez recuperaram-se sem intercorrência pós-operatória (G1) e 10 foram a óbito sete a 10 dias após a cirurgia, com sinais de choque séptico (G2). Avaliaram-se temperatura retal, freqüências cardíaca e respiratória, tempo de preenchimento capilar e teores plasmáticos das proteínas de fase aguda - fibrinogênio, ceruloplasmina, proteína C-reativa, antitripsina, haptoglobina e glicoproteína ácida -, antes e até sete dias após a laparotomia. As leucometrias às 72h e no sétimo dia pós-operatório dos eqüinos que foram a óbito foram, respectivamente, 34,6% e 57,1%, mais altas que a dos animais curados. Os maiores valores de proteína de fase aguda ocorreram no sétimo dia após a cirurgia; os percentuais de elevação de fibrinogênio, antitripsina, glicoproteina ácida, proteína C-reativa, ceruloplasmina e haptoglobina de eqüinos do G2 em relação ao G1 foram 46,8%, 67,9%, 91,9%, 112,2%, 126,9% e 186,2%, respectivamente. Twenty adult horses with acute abdomen were examined and submitted to treatment by laparotomy; ten had no postoperative complication (group 1), and ten showed septic shock symptom and died from seven to ten days after surgery (group 2). Body temperature, heart and respiratory rates, filling capillary time, and plasma acute phase protein concentrations - fibrinogen, ceruloplasmin, C-reactive protein, antitrypsin, haptoglobin, and acid glycoprotein - were evaluated before laparotomy and until seven days after surgery. White blood cell counts at 72h and seven days after surgery in group 2 animals were, respectively, 34.6% and 57.1%, and were higher than those measured in group 1 horses. The highest values of acute phase proteins occurred on the seventh day after surgery. The increase percentages of fibrinogen, antitrypsin, acid glycoprotein, C-reactive protein, ceruloplasmin, and haptoglobin plasma concentrations in group 2 animals in comparison to group 1 animals were 46.8%, 67.9%, 91.9%, 112.2%, 126.9%, and 186.2%, respectively. UNESP Faculdade de Ciências Agrárias e Veterinárias Universidade Federal do Tocantins (UFT) UNESP Faculdade de Ciências Agrárias e Veterinárias

Neste estudo, a expressão de receptores de quimiocinas na superfície dos leucócitos circulantes foi feita pela citometria de fluxo. Houve aumento da porcentagem de linfócitos CCR2+CD4+ no sangue periférico dos pacientes com hanseníase. Este resultado preliminar sugeriu alteração do perfil dos receptores de quimiocinas desses pacientes. In this study, the expression of chemokine receptors on the surface of circulating leukocytes was determined using flow cytometry. An increase in the percentage of CCR2+CD4+ lymphocytes was observed in the peripheral blood of leprosy patients. This preliminary data suggests that alterations occur in the chemokine receptor profile of these patients.

The challenge of preventing major chronic diseases requires reliable, early biomarkers. Gestational mild undernutrition in rats is enough to program the offspring to develop later pathologies; the intake of leptin, a breastmilk component, during lactation may reverse these programming effects. We used these models to identify, in peripheral blood mononuclear cells (PBMCs), transcriptomic-based early biomarkers of programmed susceptibility to later disorders, and explored their response to neonatal leptin intake. Microarray analysis was performed in PBMCs from the offspring of control and 20% gestational calorie-restricted dams (CR), and CR-rats supplemented with physiological doses of leptin throughout lactation. Notably, leptin supplementation normalised 218 of the 224 mRNA-levels identified in PBMCs associated to undernutrition during pregnancy. These markers may be useful for early identification and subsequent monitoring of individuals who are at risk of later diseases and would specifically benefit from the intake of appropriate amounts of leptin during lactation., This study was supported by the Spanish Government (grant AGL2012-33692), the European Union's Seventh Framework Programme FP72007-2013 under grant agreement n. 244995 (BIOCLAIMS Project), and the Instituto de Salud Carlos III, Centro de Investigacion Biomedica en Red Fisiopatologia de la Obesidad y Nutricion, CIBERobn. The Laboratory belongs to the Nutrigenomics-group, awarded as Group of Excellence of CAIB and supported by Direccio General d'Universitats, Recerca i Transferencia del Coneixement of Regional Government (CAIB) and FEDER funds (EU Contract: n. FP6-506360). Jadwiga Konieczna is granted with a PhD fellowship entitled beca para la formacion de personal investigador, en el marco de un programa operativo cofinanciado por el Fondo Social Europeo.

Aberrant DNA methylation pattern is a well-known epigenetic marker of cancer cells. Recently, aberrant methylation was also reported in the peripheral blood of cancer patients and it could potentially serve as a biomarker for cancer risk. We investigated the methylation pattern of LINE-1 and other repetitive DNA elements in peripheral blood of cutaneous melanoma patients in order to search for an association with clinical characteristics. The patient cohort was composed by 69 unrelated melanoma patients, 28 of whom were hereditary cases (with or without CDKN2A mutations) and 41 were isolated (sporadic) melanoma cases. Methylation of LINE-1 was evaluated by pyrosequencing, whereas additional repetitive DNA sequences were assessed using Illumina 450K methylation microarray. Melanoma patients exhibited a higher, albeit heterogeneous, LINE-1 methylation level compared with controls. Hereditary melanoma patients carrying CDKN2A mutations showed a hypermethylated pattern of both LINE-1 and repetitive DNA elements compared with other patients. In particular, the methylation level at one specific CpG of LINE-1 was found to be correlated with the occurrence of metastasis. Our data suggest that LINE-1 hypermethylation in peripheral blood of melanoma patients is a potential epigenetic biomarker for metastasis occurrence.

Cells of the mononuclear and polymorphonuclear series were analyzed throughout the course of Toxoplasma gondii infections of C57 BL/6 female mice with chronic inflammatory foci. The animals were inoculated intraperitoneally with 90-100 parasitic cysts followed by subcutaneous implantation of cover glasses (after 4 days). The shift of leukocytes from the blood to the inflammatory focus, corresponding to a prolonged infiltration of myeloid cells, was measured in a time course of 20 days. A significant decline was observed in the mature polymorphonuclear cells in the blood and a concomitant development of a marked monocytosis. Infections with T. gondii in mice with chronic inflammatory foci resulted in dedication of the entire myeloid cells system to the generation of cells that were exported to combat the inflammation, which occurred in those places, and to the chronic infection itself. The shift in the normal proportions of cells and in cells of different degrees of maturity, suggesting that a blockade in the chemokines extruded to attract polimorphonuclear cells, might account to a large extent for the failure of immune responses commonly associated with Toxoplasma subclinical infections.

Micotoxinas representam um vasto grupo de contaminantes químicos naturais originados a partir do metabolismo secundário de fungos filamentosos patogênicos. Elas são produzidas, principalmente, pelos gêneros Fusarium, Alternaria, Aspergillus e Penicillium, os quais podem contaminar grãos e cereais, como trigo, milho e soja. Conforme sua natureza e níveis de concentração, micotoxinas podem induzir efeitos tóxicos em animais de produção e humanos. Um estudo in vitro foi realizado para avaliar a susceptibilidade das células linfocitárias de frangos de corte a diferentes concentrações de ocratoxina A, deoxinivalenol e zearalenona. Cada micotoxina foi adicionada ao meio celular em diferentes concentrações (0,001; 0,01; 0,1 e 1μg/mL). A viabilidade celular e atividade de ecto-adenosina desaminase foram analisadas em 24, 48 e 72 horas através de ensaios colorimétricos. Para isso, foram utilizados 0,7x10(5) linfócitos/mL em meio RPMI 1640, suplementado com 10% de soro fetal bovino e 2,5 UI de penicilina/estreptomicina por mL, incubados em atmosfera de 5% de CO2 a 37 °C. Todos os experimentos foram realizados em triplicata e os resultados foram expressos como média e erro padrão da média. Os resultados obtidos demonstraram que tanto ocratoxina A como deoxinivalenol induziram proliferação linfocitária e baixa atividade enzimática in vitro (P0,05). Foi possível correlacionar os dados referentes à viabilidade celular e atividade de ecto-adenosina desaminase, sugerindo que, em concentrações mínimas, as micotoxinas testadas não estimularam a atividade da enzima, que possui ação pró-inflamatória e contribui para o processo de imunossupressão e, portanto, evitando um decréscimo na viabilidade celular. Este é o primeiro estudo feito com OCRA, DON e ZEA sobre linfócitos de frangos de corte em cultivos in vitro na avaliação desses parâmetros. Mycotoxins are a group of chemically diverse naturally occurring substances resulting from the secondary metabolism of pathogenic filamentous fungi. They are produced mainly by the genera Fusarium, Alternaria, Aspergillus and Penicillium which can contaminate grains and cereals such as wheat, corn and soy. According to the nature and the concentration levels, mycotoxins can induce toxic effects in food-production animals and humans. An in vitro study was conducted to evaluate the susceptibility of broiler chickens lymphocytes to different concentrations of ochratoxin A, deoxynivalenol and zearalenone. Each toxin was added to the cell medium at different concentrations (0.001, 0.01, 0.1 and 1μg/mL). Cell viability and ecto-adenosine deaminase activity were assessed at 24, 48 and 72 hours by colorimetric assays. Thus, it were used 0.7x10(5) lymphocytes/mL in RPMI 1640 medium, supplemented with 10% fetal bovine serum and 2.5 IU of penicillin/streptomycin per mL, incubated at 37°C in a 5% CO2 atmosphere. All the experiments were carried out in triplicate and the results were expressed as mean ± standard error of the mean. The results showed that OTA and DON induced lymphocyte proliferation and reduced enzymatic activity in vitro (P0,05). It was possible to correlate the results about viability cell and ecto-adenosine deaminase activity, suggesting that, at minimal concentrations, the evaluated mycotoxins do not stimulated the enzymatic activity, which has proinflammatory action and contributes for the immunosuppression process, thus, avoiding a decrease on the viability cell. This is the first in vitro study conducted with OTA, DON and ZON in broiler chickens lymphocytes evaluating these parameters.

O objetivo deste estudo foi descrever aspectos do parto e da involução uterina em ovelhas nativas do Pantanal. Foram estudadas 57 fêmeas pluríparas com cio sincronizado. Ao parto, avaliou-se sua classificação (simples ou gemelar), o tempo de expulsão da placenta, peso da placenta, total de diâmetro dos cotilédones, além do sexo e peso do cordeiro. A involução uterina foi avaliada pelo perfil leucocitário e de células de descamação (esfregaços cérvico-uterinos) e pela ultrassonografia uterina transretal nos dias 1, 7, 14, 21 e 28 dias pós-parto. O peso da placenta foi superior (P0.05) between sex and parturition classification. The total of neutrophils in twin parturitions were higher in days 1, 7 and 14 postpartum (78.5±9.5; 58.5±9.6 and 31.9±9.8, respectively). Macrophages were different (P

Although resting immune function is not very different in athletes compared with non-athletes periods of intensified training (overreaching) in already well trained athletes can result in a depression of immunity in the resting state. Illness-prone athletes appear to have an altered cytokine response to antigen stimulation and exercise. Having low levels of salivary IgA secretion also makes athletes more susceptible to upper respiratory tract infections. Overtraining is associated with recurrent infections and immunodepression is common, but immune functions do not seem to be reliable markers of impending overtraining. There are several possible causes of the diminution of immune function associated with periods of heavy training. One mechanism may simply be the cumulative effects of repeated bouts of intense exercise (with or without tissue damage) with the consequent elevation of stress hormones, particularly glucocorticoids such as cortisol, causing temporary inhibition of TH-1 cytokines with a relative dampening of the cell-mediated response. When exercise is repeated frequently there may not be sufficient time for the immune system to recover fully. Tapering has been described as a gradual reduction in the training load which allows the recovery of physiological capacities that were impaired by previous intensive training and permits further training-induced adaptations to occur accompanied by competition performance enhancements. The majority of the studies that have examined the recovery of immunoendocrine responses during 1-3 week tapers in trained athletes have mainly reported enhanced performance, often accompanied by increased anabolic activity, reduced physiological stress and restoration of mucosal immunity and immune function.Quando se compara a função imune, em repouso, de atletas e não atletas, não se verificam grandes diferenças. Porém, períodos de treinamento intensificado ("overreaching") em atletas bem treinados podem induzir supressão da imunidade no estado de repouso. Os atletas com maior propensão para contrair doenças parecem apresentar uma resposta alterada de citocinas, tanto quando estas são estimuladas por antígenos, quanto em resposta ao exercício propriamente dito. Baixos níveis de secreção de IgA salivar também contribuem para tornar os atletas mais susceptíveis à infecções do trato respiratório superior. A síndrome do "overtraining" é associada a infecções recorrentes e a imunossupressão é comum; no entanto, marcadores da função imune não parecem ser suficientemente sensíveis ao "overtraining" eminente. Existem várias possíveis causas para a diminuição da função imune associadas com períodos de treinamento severo. Um possível mecanismo pode ser simplesmente, o efeito acumulativo de atividades e sessões repetidas de exercício intenso (com ou sem dano tecidual), com a consequente elevação dos hormônios de estresse, particularmente os glicocorticóides como o cortisol, causando assim, uma inibição temporária das citocinas de TH-1, com uma relativa atenuação da resposta imune celular. Quando o exercício é repetido frequentemente, pode não haver tempo suficiente para uma total recuperação do sistema imunológico. O "Tapering" tem sido descrito como uma gradual redução na carga de treinamento a qual permite a recuperação das capacidades fisiológicas, que por sua vez, foram afetadas pelo treinamento intensivo anterior, permitindo assim, que adaptações adicionais decorrentes do treinamento ocorram, acompanhadas pelo incremento do desempenho competitivo. A maioria dos estudos que investigaram a recuperação das respostas imuno-endócrinas em atletas durante uma a três semanas de "taper" tem registrado aumento do desempenho, frequentemente acompanhado pelo aumento da atividade anabólica, redução do estresse fisiológico e restabelecimento da imunidade das mucosas e da função imune.

The aims of the present study were to explore the expression pattern of haem biosynthesis enzymes in circulating cells of patients affected by two types of porphyria (acute intermittent, AIP, and variegate porphyria, VP), together with the antioxidant enzyme pattern in AIP in order to identify a possible situation of oxidative stress. Sixteen and twelve patients affected by AIP and VP, respectively, were analysed with the same numbers of healthy matched controls. Erythrocytes, neutrophils and peripheral blood mononuclear cells (PBMCs) were purified from blood, and RNA and proteins were extracted for quantitative real time PCR (qRT-PCR) and Western-blot analysis, respectively. Porhobilinogen deaminase (PBGD) and protoporphyrinogen oxidase (PPOX) gene and protein expression was analysed. Antioxidant enzyme activity and gene expression were additionally determined in blood cells, together with protein carbonyl content in plasma. PBMCs isolated from AIP patients presented low mRNA levels of PBGD when compared to controls, while PBMCs isolated from VP patients presented a decrease in PPOX mRNA. PPOX protein content was higher in AIP patients and lower in VP patients, compared to healthy controls. Regarding antioxidant enzymes, PBMCs and erythrocyte superoxide dismutase (SOD) presented statistically significant higher activity in AIP patients compared to controls, while catalase activity tended to be lower in these patients. No differences were observed regarding antioxidant gene expression in white blood cells. Circulating cells in AIP and VP patients present altered expression of haem biosynthetic enzymes, which could be useful for the differential diagnosis of these two types of porphyria in certain difficult cases. AIP patients present a condition of potential oxidative stress similar to VP patients, evidenced by the post-transcriptional activation of SOD and possible catalase impairment., This work has been granted by Accion Estrategica en Salud del Ministro de Ciencia e Innovacion (DPS2008-07033-C03-03), Programme of Promotion of Biomedical Research and Health Sciences, Projects 11/01791, Red Predimed-RETIC RD06/0045/1004, CIBERobn CB12/03/30038, Balearic Island Government and FEDER funds (35/2011 and 23/2012), PROMETEO/2012/007 from Generalitat Valenciana to ER. ER is recipient of Institute de Salud Carlos III-FEDER (PS09/01093) and Fundacion Salud 2000-Merck Serono grants. LC-Q was recipient of a CONACYT-Mexico (ref 197139) fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.; This work has been granted by Accion Estrategica en Salud del Ministro de Ciencia e Innovacion (DPS2008-07033-C03-03), Programme of Promotion of Biomedical Research and Health Sciences, Projects 11/01791, Red Predimed-RETIC RD06/0045/1004, CIBERobn CB12/03/30038, Balearic Island Government and FEDER funds (35/2011 and 23/2012), PROMETEO/2012/007 from Generalitat Valenciana to ER. ER is recipient of Instituto de Salud Carlos IIIFEDER (PS09/01093) and Fundacion Salud 2000-Merck Serono grants. LC-Q was recipient of a CONACYT-Mexico (ref 197139) fellowship.

Background and aimsSchistosomiasis is an intravascular parasitic disease associated with inflammation. Endothelial cells control leukocyte transmigration and vascular permeability being modulated by pro-inflammatory mediators. Recent data have shown that endothelial cells primed in vivo in the course of a disease keep the information in culture. Herein, we evaluated the impact of schistosomiasis on endothelial cell-regulated events in vivo and in vitro.Methodology and principal findingsThe experimental groups consisted of Schistosoma mansoni-infected and age-matched control mice. In vivo infection caused a marked influx of leukocytes and an increased protein leakage in the peritoneal cavity, characterizing an inflamed vascular and cellular profile. In vitro leukocyte-mesenteric endothelial cell adhesion was higher in cultured cells from infected mice as compared to controls, either in the basal condition or after treatment with the pro-inflammatory cytokine tumor necrosis factor (TNF). Nitric oxide (NO) donation reduced leukocyte adhesion to endothelial cells from control and infected groups; however, in the later group the effect was more pronounced, probably due to a reduced NO production. Inhibition of control endothelial NO synthase (eNOS) increased leukocyte adhesion to a level similar to the one observed in the infected group. Besides, the adhesion of control leukocytes to endothelial cells from infected animals is similar to the result of infected animals, confirming that schistosomiasis alters endothelial cells function. Furthermore, NO production as well as the expression of eNOS were reduced in cultured endothelial cells from infected animals. On the other hand, the expression of its repressor protein, namely caveolin-1, was similar in both control and infected groups.Conclusion/significanceSchistosomiasis increases vascular permeability and endothelial cell-leukocyte interaction in vivo and in vitro. These effects are partially explained by a reduced eNOS expression. In addition, our data show that the disease primes endothelial cells in vivo, which keep the acquired phenotype in culture.

O presente estudo comparou os níveis plasmáticos de glicose, proteínas totais, uréia, triglicerídeos e colesterol, a contagem de eritrócitos, leucócitos e trombócitos entre alevinos e juvenis de Arapaima gigas (Osteoglociformes, Arapaimidae) de uma piscicultura semi-intensiva de Manaus, estado do Amazonas, Brasil. Os alevinos de A. gigas apresentaram níveis significativamente (p0,05) no número de trombócitos e leucócitos totais entre alevinos e juvenis. Os juvenis de A. gigas apresentaram menor quantidade de linfócitos e maior quantidade de monócitos, neutrófilos e eosinófilos, quando comparados aos alevinos. Em estudos futuros, espécimes de A. gigas com outras idades também deverão ser comparados para melhor entendimento dos efeitos da idade neste peixe amazônico, pois estas informações, além de possibilitarem uma melhor compreensão da fisiologia desta espécie, poderão auxiliar no estabelecimento de estratégias para seu manejo. The present study compared the biochemical and hematological parameters of fingerlings and juveniles Arapaima gigas (Osteoglociformes, Arapaimidae) from a semi-intensive fish farming in Manaus, Amazon State, Brazil. Fingerlings A. gigas had lower levels of total protein, triglycerides, urea and mean cell volume (MCV), but higher (p0.05) in the total thrombocytes and total white blood cell counts between fingerlings and juveniles. Juveniles A. gigas presented lower lymphocytes number and higher monocytes, neutrophils and eosinophils number when compared to the fingerlings. In further studies, A. gigas specimens with other ages should also be compared for a better understanding of the ontogenic development in this Amazon fish, because this information besides increase the knowledge of the physiology of this species, can aid in the establishment of strategies for its adequate handling.

Foram utilizados 64 leitões, submetidos a oito tratamentos: ração com leite em pó (LP), três rações com níveis crescentes de plasma suíno (PS), três rações com níveis crescentes de ovo inteiro (OI) e ração com alto nível de farelo de soja (FS). Foi coletado sangue dos animais aos 27 e 34 dias de idade para avaliação dos parâmetros sangüíneos, e aos 28 e 35 dias de idade quatro animais por tratamento foram abatidos para coleta do pâncreas e posterior determinação do peso absoluto (PA) e peso relativo (PR) do pâncreas e da atividade da tripsina (AT). Os tratamentos não influenciaram significativamente o PA e a AT. Verificou-se efeito significativo dos níveis crescentes de PS, com redução linear dos leucócitos e aumento do volume globular, aos 27 dias; enquanto aos 34 dias, observou-se aumento linear das hemácias. Aos 27 dias, os animais arraçoados com níveis de PS apresentaram menor percentagem de eosinófilos do que aqueles arraçoados com níveis de OI. A utilização de plasma suíno promoveu menor estímulo à resposta imune, enquanto que a utilização de ovo inteiro promoveu maior resposta humoral dos leitões. Were used 64 piglets submitted eight treatments: ration with skim milk (SM), three rations with crescent levels of swine plasma (SP), three rations with whole egg (WE) and a ration with high inclusion of soybean meal (SB). Were monitored the blood parameters (BP) in pigs at 27 and at 34 days of age. The piglets were slaugther at 28 and at 35 days of age, for collections pancreas and posterior mensurements of absolut (AW) and relative weigth (RW) of pancreas and trypsin activity (TA). Treatments not influencied AW and TA. Significant effect of the crescent levels was verified of SP, with lineal reduction of the leukocytes and increase of the globular volume, to the 27 days; while to the 34 days, lineal increase of the hematias was observed. At 27 days, animals feds rations with crescent levels of SP have inferior percentage of eosinophils than others that consumed crescent levels of WE. The utilization of SP promoted smaller stimulus to the immune reply, while the use of WE promoted larger humoral reply of the piglets.

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

OBJETIVO: Avaliar a importância da interação entre a integrina Mac-1 dos leucócitos (a Mb 2) e a glicoproteína (GP) Iba das plaquetas para o recrutamento de leucócitos após a lesão vascular e o efeito da neutralização da interação Mac-1-GPIba sobre a proliferação celular e a hiperplasia neointimal desencadeadas por lesão vascular. MÉTODOS: Um peptídeo denominado M2 ou anticorpo anti-M2 foi desenvolvido para bloquear a interação Mac-1-GPIba . Esse peptídeo foi injetado e comparado com anticorpo-controle em camundongos C57B1/6J submetidos a lesão vascular da artéria femoral com corda-guia. Um, cinco ou 28 dias após a lesão vascular, as artérias femorais foram retiradas para a realização de morfometria e imuno-histoquímica. RESULTADOS: O bloqueio da interação Mac-1-GPIba promoveu uma redução estatisticamente significativa do número de leucócitos na camada média no primeiro dia após a lesão vascular (controle: 7,9±5,0% do total de células versus anti-M2: 2,0±1,6%, p=0,021), bem como determinou uma diminuição estatisticamente significativa do acúmulo de leucócitos na neoíntima em cinco e 28 dias (controle: 42,3±12,9% versus anti-M2: 24,6±10,8%, p=0,047 e controle: 7,9±3,0% versus anti-M2: 3,3±1,3%, p=0,012; respectivamente). A proliferação celular na camada média do vaso em cinco dias pós-lesão foi reduzida com o bloqueio da interação Mac-1-GPIba (controle: 5,0±2,9% do total de células versus anti-M2: 1,8±0,5%; p=0,043), assim como houve diminuição significativa da proliferação celular na camada íntima do vaso em 28 dias (controle: 3,8±1,7% versus anti-M2: 2,0±1,2%; p=0,047). O bloqueio da interação Mac-1-GPIba também determinou uma redução estatisticamente significativa do espessamento intimal em 28 dias pós-lesão (controle: 10.395±3.549 µm² versus anti-M2: 4.561±4.915 µm²; p=0,012). CONCLUSÃO: O recrutamento de leucócitos após a lesão vascular é dependente da interação Mac-1-GPIba e a neutralização dessa interação inibe a proliferação celular e a formação neointimal. OBJECTIVE: To assess the importance of the interaction between leukocyte integrin Mac-1 (a Mb 2) and platelet glycoprotein (GP) Ib-a for leukocyte recruitment after vascular injury and the effect of the neutralization of the Mac-1-GPIba interaction on cell proliferation and the neointimal hyperplasia triggered by the vascular injury. METHODS: A peptide called M2 or anti-M2 antibody was developed to block the Mac-1-GPIba interaction. This peptide was injected and compared to a control-peptide in C57B1/6J mice submitted to vascular injury of the femoral artery with a guide wire. One, five or 28 days after the vascular injury, the femoral arteries were removed for morphometric and immunohistochemical analyses. RESULTS: The blocking of the Mac-1-GPIba interaction promoted a statistically significant reduction in the number of leukocytes in the neointimal layer on the first day after the vascular injury (control: 7.9±5.0% of the cell total versus anti-M2: 2.0±1.6%, p=0.021), as well as determined a statistically significant decrease in leukocyte accumulation in the neointimal layer on days 5 and 28 (control: 42.3±12.9% versus anti-M2: 24.6±10.8%, p=0.047 and control: 7.9±3.0% versus anti-M2: 3.3±1.3%, p=0.012; respectively). Cell proliferation in the neointimal layer of the vessel five days post-injury was reduced with the blocking of the Mac-1-GPIba interaction (control: 5.0±2.9% of the cell total versus anti-M2: 1.8±0.5%; p=0.043), along with a significant decrease in cell proliferation in the vessel neointimal layer 28 days post-injury (control: 3.8±1.7% versus anti-M2: 2.0±1.2%; p=0.047). The blocking of the Mac-1-GPIba interaction also determined a statistically significant decrease of the intimal thickening 28 days post-injury (control: 10,395±3,549 µm² versus anti-M2: 4,561±4,915 µm²; p=0.012). CONCLUSION: Leukocyte recruitment after a vascular injury depends on the Mac-1-GPIba interaction and the neutralization of this interaction inhibits cell proliferation and neointimal formation.

The aim of this paper is to compare three different methods for counting white blood cells [WBC] (Natt and Herrick method, estimation with 1,000 and 2,000 erythrocytes) and three methods for counting total thrombocytes [TT] (Wojtaszek method, estimation with 1,000 and 2,000 erythrocytes) in a South American freshwater turtle species, Podocnemis expansa, Schweigger 1812 (Reptilia, Pelomedusidae). Direct WBC counts using the Natt and Herrick method showed limitations, which are discussed here. The WBC and TT counts using 1,000 erythrocytes from blood smears are not recommended for Amazon turtles nor other reptilian species, since wide variation in counts can be observed. Estimation methods for determining WBC and TT based on 2,000 erythrocytes of blood smears were most acceptable because they allow a differentiation between leukocytes and thrombocytes and also had a smaller variation. The methods investigated here for the Amazon turtle, which have been widely used in other reptile species, provided evidence that the most acceptable method is not that of using diluted stains and a hemocytometer. O objetivo deste estudo foi comparar três diferentes métodos para contar leucócitos totais [LT] (método de Natt & Herrick, de estimação em 1000 e 2000 eritrócitos) e três métodos para contar trombócitos totais [TT] ( método de Wojtaszek, de estimação em 1000 e 2000 eritrócitos) em uma espécie de tartaruga de água doce da América do Sul, Podocnemis expansa, Schweigger 1812 (Reptilia, Pelomedusidae). As contagens diretas de LT usando o método de Natt & Herrick mostraram limitações que são aqui discutidas. As contagens de LT e TT usando estimativa em 1000 eritrócitos na extensão sanguínea não são recomendadas para tartaruga-da-Amazônia nem para outras espécies de répteis, pois houve ampla variação nestas contagens. Os métodos para determinar LT e TT baseados em 2000 eritrócitos nas extensões sanguíneas foram mais aceitáveis porque eles permitem uma diferenciação entre leucócitos e trombócitos, além disso, teve uma variação menor. Os métodos aqui investigados para tartaruga-da-Amazônia, os quais são amplamente usados em outras espécies de répteis, proveram evidências de que o método mais aceitável não é o que usa corantes diluentes e um hemocitômetro.

Introduction: It has been suggested that individuals exposed to Mycobacterium tuberculosis can eliminate the infection through the innate immune response without the participation of the acquired immunity. However, T helper type 1 immune response is critical to control persistent infection. Objective: The purpose of this study was to evaluate the distribution of leukocyte and lymphocyte subsets in peripheral blood of individuals with pulmonary tuberculosis (TB) and positive Tuberculin Skin Test (TST). Methodology: In order to achieve that goal, we have collected blood sample from thirty-four treatment-naïve TB patients, fifty TST-positive and forty-one TST-negative individuals. Results: The evaluation has shown a significant reduction in the number of total lymphocytes, B cells, CD4+ and CD8+ T cells and percentage increase of NK cells, neutrophils and monocytes in TB patients, when compared to TSTpositive and TST-negative individuals. There was no statistical difference for leucocyte and lymphocyte subsets between TST-positive and TST-negative groups. Peripheral blood white cell counts change significantly at diagnosis. Conclusion: The quantification of these cells may support the diagnosis and monitoring of patients with pulmonary tuberculosis.

Os mecanismos envolvidos no processo de reparo de tecidos estão revisados nesse trabalho. O processo de cicatrização ocorre fundamentalmente em três fases: inflamação, formação de tecido de granulação e deposição de matriz extracelular e remodelação. Os eventos celulares e tissulares de cada uma dessas fases estão descritos e discutidos. Os mediadores químicos estão correlacionados com os eventos do processo de cicatrização e as células envolvidas. Especial ênfase é dada à participação dos fatores de crescimento. The mechanisms involved in tissue repair are revised. The wound healing process occurs basically in three phases: inflammation, formation of granulating tissue and extracellular tissue deposition, and tissue remodeling. The cellular and tissue events of each phase are described and discussed. The chemical mediators and their interplay with the wound healing events and cells involved are also discussed. However, especial attention was given to the role played by the growth factors in the tissue repair process.