Avaliação imunofenotípica de subpopulações linfocitárias no sangue do cordão umbilical e periférico de suínos neonatos (Sus scrofa)

Considering the importance of umbilical cord blood as a potential source of stem cell and, on the other hand, the use of the domestic swine (Sus scrofa) as a useful model for biomedical research in regenerative medicine and aiming to contribute about the quantification of lymphocyte subsets in umbilical cord blood and peripheral blood of newborn piglets, this study aimed to quantify CD4+, CD5+ and CD8+ cells from umbilical cord blood and peripheral blood from pigs at term blood samples. Were analyzed samples of the umbilical cord blood and peripheral of 48 piglets of Topigs lineage, from healthy mothers, artificially inseminated and natural birth. Blood samples were collected from the umbilical cord at birth, by the umbilical vein, and peripheral blood by venous sinus retro-ophthalmic. The immunological measurements of CD4+, CD5+ and CD8+ were obtained by flow cytometry. The relative average values for the CD4+, CD5+ e CD8+ counts in umbilical cord blood and peripheral blood of newborn piglets were inferior to those reported for peripheral blood in adult pigs, suggesting immunological immaturity. The ratio CD4+:CD8+ in umbilical cord blood (3.2±1.2%) and peripheral blood (3.2±1.7%) showed a predominance of TCD4+ over TCD8+. The percentage of CD4+ and CD8+ cells was 1.37±0.86% and 1.15±0.57%, respectively, in umbilical cord blood and peripheral blood.

Ações antioxidantes e imunológicas do selênio em cães

Pós-graduação em Zootecnia - FMVZ

Analysis of Toll-Like Receptors, iNOS and Cytokine Profiles in Patients with Pulmonary Tuberculosis during Anti-Tuberculosis Treatment

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Atividade imunomoduladora de parasporina de Bacillus thuringiensis sobre linfócitos e monócitos humanos

Bacillus thuringiensis is an environmental bacteria that produces a group of crystallizable proteins (Cry) that are toxic for several insects and worms species. Recently, it was described a novel class of Cry proteins called parasporins (PS) that showed cytotoxic effects on animal and human tumor cells. Six types of PS have been described so far, PS1 to PS6, and their cytotoxic activity has been studied. However, the direct effect on tumor cells has been the current research focus, while the immunomodulatory role of the PS has not been studied yet. Therefore, this study aimed to verify whether PS of TC 2.3.1R6 B. thuringiensis strain has immunostimulatory activity on human lymphocytes and monocytes. We have evaluated the protein toxicity against human cells, the lymphoproliferative activity and the effects on peripheral blood monocytes. The PS-PK showed no toxic or stimulating activity on lymphocyte proliferation. However, it inhibited the spontaneous production of IL-10 as well as ConA-induced and the production of IFN-γ. PS-PK decreased the release of hydrogen peroxide and increased the production of TNF- α by monocytes. PS-PK performed inhibitory production of hydrogen peroxide and TNF-α by monocytes, whereas PS-Tp showed stimulation of the production of hydrogen and TNF-α

Células T reguladoras em sangue periférico de cães portadores de mastocitoma cutâneo

Pós-graduação em Medicina Veterinária - FCAV

Toxicidade do cloridrato de doxorrubicina na dose de 120mg/m², em cadelas com tumor venéreo transmissível

Pós-graduação em Cirurgia Veterinária - FCAV

Imunofenótipos de linfócitos T no sangue e no liquor de cães com leishmaniose visceral: correlação com as lesões encefálicas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The involvement of TLR2 and TLR4 in cytokine and nitric oxide production in visceral leishmaniasis patients before and after treatment with anti-leishmanial drugs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

IRS-1 gene polymorphism and DNA damage in pregnant women with diabetes or mild gestational hyperglycemia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Metabolic activity of the sticker's lymphosarcoma

1. 1. Some parameters (glycolysis, respiration, levels of glycolytic enzymes) of the lymphoid cells from the Sticker's lymphosarcoma were established in order to better define the biochemical behavior of the venereal tumor of the dog. 2. 2. For comparative purposes lymphocytes from peripheral blood of normal tumor-bearing dogs were also studied. 3. 3. Lactic acid produced by the tumor cells during aerobic glycolysis is liberated in the reaction medium. 4. 4. Oxygen uptake is enhanced in the presence of succinate, but not with pyruvate, α-ketoglutarate, or malate as substrates. 5. 5. Higher levels of some of the enzymes from the glycolytic pathways as well as differences on the physicochemical and kinetic properties of the glycolytic regulatory enzymes are found in Sticker's tumor cells, when compared with the lymphocytes from peripheral blood of normal and tumor-bearing dogs. 6. 6. A fructose-bisphosphate positively modulated pyruvatekinase is found in the tumor cells. © 1987.

Activation of monocytes and cytokine production in patients with peripheral atherosclerosis obliterans

Background: Arterial peripheral disease is a condition caused by the blocked blood flow resulting from arterial cholesterol deposits within the arms, legs and aorta. Studies have shown that macrophages in atherosclerotic plaque are highly activated, which makes these cells important antigen-presenting cells that develop a specific immune response, in which LDLox is the inducing antigen. As functional changes of cells which participate in the atherogenesis process may occur in the peripheral blood, the objectives of the present study were to evaluate plasma levels of anti-inflammatory and inflammatory cytokines including TNF-alpha, IFN-gamma, interleukin-6 (IL-6), IL-10 and TGF-beta in patients with peripheral arteriosclerosis obliterans, to assess the monocyte activation level in peripheral blood through the ability of these cells to release hydrogen peroxide (H(2)O(2)) and to develop fungicidal activity against Candida albicans (C. albicans) in vitro.Methods: TNF-alpha, IFN-gamma, IL-6, IL-10 and TGF-beta from plasma of patients were detected by ELISA. Monocyte cultures activated in vitro with TNF-alpha and IFN-gamma were evaluated by fungicidal activity against C. albicans by culture plating and Colony Forming Unit (CFU) recovery, and by H(2)O(2) production.Results: Plasma levels of all cytokines were significantly higher in patients compared to those detected in control subjects. Control group monocytes did not release substantial levels of H(2)O(2) in vitro, but these levels were significantly increased after activation with IFN-gamma and TNF-alpha. Monocytes of patients, before and after activation, responded less than those of control subjects. Similar results were found when fungicidal activity was evaluated. The results seen in patients were always significantly smaller than among control subjects. Conclusions: The results revealed an unresponsiveness of patient monocytes in vitro probably due to the high activation process occurring in vivo as corroborated by high plasma cytokine levels.

Human gammaherpesviruses viraemia in HIV infected patients

The Epstein-Barr virus (EBV) and Kaposi's sarcoma associated herpesvirus (KSHV) are consistently associated with lymphoproliferative diseases and cancers in humans, notably in patients with HIV. Our aim was to evaluate whether EBV and/or KSHV viral loads regularly assessed in peripheral blood mononuclear cells (PBMC) correlate with clinical or laboratorial parameters retrieved for patients living with HIV. This was a longitudinal study with a cohort of 157 HIV positive patients attending an academic HIV outpatient clinic in São Paulo State, Brazil. For each patient, up to four blood samples were collected over a 1 year clinical follow-up: on enrolment into the study, and after 4, 8 and 12 months. Total DNA was extracted from PBMC, and EBV and KSHV viral loads were assessed by real time quantitative PCR. Higher viral loads for EBV were significantly associated with high HIV viraemia, a greater number of circulating T CD8+ cells and lack of virological response to the antiretroviral treatment. KSHV viral load was undetectable in virtually all samples. EBV viral load in PBMC correlated with the number of circulating T CD8+ lymphocytes and the response to the antiretroviral therapy in HIV infected patients. In contrast, KSHV was undetectable in PBMC, presumably an effect of the antiretroviral treatment. Therefore, either KSHV infection in the population studied was absent or viral load in PBMC was beyond the analytical limit of the assay.

Increased DNA damage is related to maternal blood glucose levels in the offspring of women with diabetes and mild gestational hyperglycemia

This study aimed at correlating maternal blood glucose levels with DNA damage levels in the offspring of women with diabetes or mild gestational hyperglycemia (MGH). Based on oral glucose tolerance test results and glycemic profiles, 56 pregnant women were allocated into 3 groups: nondiabetes, MGH, and diabetes. The offspring of these women (56 infants) were also evaluated. Maternal peripheral blood and umbilical cord blood samples were collected and processed for biochemical and DNA damage analysis by the comet assay. A positive correlation between maternal blood glucose mean and increased offspring DNA damage levels was observed. Hyperglycemia played a role in offspring DNA damage, but other diabetes-induced complications were also involved. Increased maternal blood glucose levels can lead to increased offspring DNA damage levels. Therefore, the monitoring, control, and treatment of pregnant women with diabetes and MGH are highly important to ensure a risk-free pregnancy and healthy infants.

Whole blood genome-wide gene expression profile in males after prolonged wakefulness and sleep recovery

Pellegrino R, Sunaga DY, Guindalini C, Martins RC, Mazzotti DR, Wei Z, Daye ZJ, Andersen ML, Tufik S. Whole blood genome-wide gene expression profile in males after prolonged wakefulness and sleep recovery. Physiol Genomics 44: 1003-1012, 2012. First published September 4, 2012; doi: 10.1152/physiolgenomics.00058.2012.-Although the specific functions of sleep have not been completely elucidated, the literature has suggested that sleep is essential for proper homeostasis. Sleep loss is associated with changes in behavioral, neurochemical, cellular, and metabolic function as well as impaired immune response. Using high-resolution microarrays we evaluated the gene expression profiles of healthy male volunteers who underwent 60 h of prolonged wakefulness (PW) followed by 12 h of sleep recovery (SR). Peripheral whole blood was collected at 8 am in the morning before the initiation of PW (Baseline), after the second night of PW, and one night after SR. We identified over 500 genes that were differentially expressed. Notably, these genes were related to DNA damage and repair and stress response, as well as diverse immune system responses, such as natural killer pathways including killer cell lectin-like receptors family, as well as granzymes and T-cell receptors, which play important roles in host defense. These results support the idea that sleep loss can lead to alterations in molecular processes that result in perturbation of cellular immunity, induction of inflammatory responses, and homeostatic imbalance. Moreover, expression of multiple genes was downregulated following PW and upregulated after SR compared with PW, suggesting an attempt of the body to re-establish internal homeostasis. In silico validation of alterations in the expression of CETN3, DNAJC, and CEACAM genes confirmed previous findings related to the molecular effects of sleep deprivation. Thus, the present findings confirm that the effects of sleep loss are not restricted to the brain and can occur intensely in peripheral tissues.

High Rates of Detection of Respiratory Viruses in Tonsillar Tissues from Children with Chronic Adenotonsillar Disease

Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR) in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05), and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05) in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.