Imunidade celular em caninos neonatos - do nascimento ao 45° dia de idade

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

Non-specific immune parameters and physiological response of Nile tilapia fed beta-glucan and vitamin C for different periods and submitted to stress and bacterial challenge

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

Alterações hematológicas em cães e gatos sob estresse

The leukocyte count interpretation on the blood helps on understanding about the possible dysfunction showed by the animal. In general, an abnormal leukogram allows the identification of pathological process. A lot of events lead to an increase in the number of neutrophils, among then, inflammatory process, infection and stress. Dependent on the stress mediator that the animal is submited, the leukocyte alteration will be different. Is important to know how the stress acts on the animal organism, changing the hematological parameters, in order to avoid mistakes on diseases‟ diagnosis. It‟s known that after a stress situation, it‟s expected that stress leukogram appears on the hemogram of the animal. However there are different kinds of stress with different responses. It‟s believed that when the animal is under an acute stress situation, a clinical picture of physiologic leukocytosis occurs which is mediated by the epinephrine. This is characterized by neutrophilia, lymphocytosis, monocytosis and eosinofilia. The physiologic leukocytosis is a transient alteration in the leukogram that occurs within minutes of the stimulus and due to fleeting effect of catecolamines it resolves within 20 to 30 minutes. However, if the animal is under chronicle stress, as solitude, it „s expected to find the stress leukogram on the leukogram, that can be noted especially in dogs. The stress leukogram is characterized by leukocytosis, neutrophilia, lymphopenia, monocytosis and eosinopenia. This kind of stress is induced by the glucocorticoids. It´s important to understand that stress leukogram takes time to occur. The variation on time depends on the animal species, although, in general, the glucocorticoides have a peak effects among 4 to 8 hours, which can last from 24 hours to 2 or 3 days

Leucograma e perfil bioquímico sérico de cabras com mastite experimental causada por Staphylococcus aureus e Streptococcus agalactiae

Pós-graduação em Medicina Veterinária - FCAV

Perfil hemostático e hematológico de equinos com compactação de cólon maior submetidos ao tratamento medicamentoso

Pós-graduação em Medicina Veterinária - FCAV

Evaluation of zebu nellore cattle blood samples using the Cell-Dyn 3500 hematology analyzer

The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Cell-dyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510) and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson's correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semi-automated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

The immune system is limited by oxidative stress: dietary selenium promotes optimal antioxidative status and greatest immune defense in pacu Piaractus mesopotamicus

Reactive oxygen species (ROS) are reactive molecules containing oxygen, that form as byproducts of aerobic metabolism, including immune system processes. Too much ROS may cause oxidative stress. In this study, we examined whether it can also limit the production of immune system compounds. To assess the relationship between antioxidant status and immunity we evaluated the effect of dietary supplementation with organic selenium, given at various levels for 10 days, on the antioxidant and immune system of the pacu fish (Piaractus mesopotamicus). Fish fed a diet containing 0.6 mg Se-yeast kg(-1) showed significant improvement in antioxidant status, as well as in hematological and immunological profiles. Specifically, they had the highest counts for catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), red blood cells, and thrombocytes; the highest leukocyte count (particularly for monocytes); and the highest serum lysozyme activity. There was also a positive correlation between GPx and lysozyme in this group of fish. These findings indicate that short-term supplementation with 0.6 mg Se-yeast kg(-1) reestablished the antioxidative status, allowing the production of innate components which can boost immunity without the risk of oxidative stress. This study shows a relationship between oxidative stress and immunity, and, from a practical perspective, shows that improving immunity and health in pacu through the administration of selenium could improve their growth performance.

Leukograms of healthy Holstein calves within the first month of life

To establish reference values and to assess the influence of age on the leukograms of healthy Holstein calves, blood samples were obtained from 300 animals. These samples were distributed equally (n=20) among 15 experimental groups according to age: birth to 8 hours, 9 to 16 hours, 17 to 24 hours, 2 days, 3 days, 4 days, 5 days, 6 to 7 days, 8 to 9 days, 10 to 11 days, 12 to 13 days, 14 to 15 days, 16 to 20 days, 21 to 25 days and 26 to 30 days of age. The maximum numbers of leukocytes (9,305.0/mL), segmented neutrophils (6,551.2/mL) and total neutrophils (6,678.3/mL) were noted within the first 8 hours of life, while band neutrophils peaked in number (133.3/mL) between 9 and 16 hours after birth. Meanwhile, the maximum total lymphocyte (4,992.1/µL) and typical lymphocyte (4,686.1/µL) counts occurred between 21 and 25 days, whereas atypical lymphocytes (388.5/µL) reached their maximum number between 26 and 30 days, demonstrating an inversion of the neutrophil:lymphocyte ratio over time. Thus, the influence of age on the leukocyte count of the evaluated calves was verified. The release of endogenous corticosteroids during labor or at birth may contribute to this variation in leukograms with age.

Impact of FLT3 Internal Tandem Duplication on the Outcome of Related and Unrelated Hematopoietic Transplantation for Adult Acute Myeloid Leukemia in First Remission: A Retrospective Analysis

Purpose Patients with acute myeloid leukemia (AML) and FLT3/internal tandem duplication (FLT3/ITD) have poor prognosis if treated with chemotherapy only. Whether this alteration also affects outcome after allogeneic hematopoietic stem-cell transplantation (HSCT) remains uncertain. Patients and Methods We analyzed 206 patients who underwent HLA-identical sibling and matched unrelated HSCTs reported to the European Group for Blood and Marrow Transplantation with a diagnosis of AML with normal cytogenetics and data on FLT3/ITD (present: n = 120, 58%; absent: n = 86, 42%). Transplantations were performed in first complete remission (CR) after myeloablative conditioning. Results Compared with FLT3/ITD-negative patients, FLT3/ITD-positive patients had higher median leukocyte count at diagnosis (59 v 21 x 10(9)/L; P < .001) and shorter interval from CR to transplantation (87 v 99 days; P = .04). Other characteristics were similar in the two groups. At 2 years, relapse incidence (RI; +/- standard deviation) was higher (30% +/- 5% v 16% +/- 5%; P = .006) and leukemia-free survival (LFS) lower (58% +/- 5% v 71% +/- 6%; P = .04) in FLT3/ITD-positive compared with FLT3/ITD-negative patients. In multivariate analyses, FLT3/ITD led to increased RI (hazard ratio [HR], 3.4; 95% CI, 1.46 to 7.94; P = .005), as did older age, female sex, shorter interval between CR and transplantation, and higher number of chemotherapy courses before achieving CR. FLT3/ITD positivity was associated with decreased LFS (HR, 0.37; 95% CI, 0.19 to 0.73; P = .002), along with older age and higher number of chemotherapy courses before achieving CR. Conclusion FLT3/ITD adversely affected the outcome of HSCT in the same direction it does after chemotherapy; despite this, more than half of the patients harboring this mutation who received transplants were alive and leukemia free at 2 years. To further improve the results, use of FLT3 inhibitors before or after HSCT deserves investigation.

The protective effect of Canova homeopathic medicine in cyclophosphamide-treated non-human primates

Background: Canova activates macrophages and indirectly induces lymphocyte proliferation. Here we evaluated the effects of Canova in cyclophosphamide-treated non-human primates. Methods: Twelve Cebus apella were evaluated. Four animals were treated with Canova only. Eight animals were treated with two doses of cyclophosphamide (50 mg/kg) and four of these animals received Canova. Body weight, biochemistry and hematologic analyses were performed for 40 days. Micronucleus and comet assays were performed for the evaluation of DNA damage. Results: We observed that cyclophosphamide induced abnormal WBC count in all animals. However, the group treated with cyclophosphamide plus Canova presented a higher leukocyte count than that which received only cyclophosphamide. Cyclophosphamide induced micronucleus and DNA damage in all animals. The frequency of these alterations was significantly lower in the Canova group than in the group without this medicine. Conclusions: Our results demonstrated that Canova treatment minimizes cyclophosphamide myelotoxicity in C. apella. (C) 2012 Elsevier Ltd. All rights reserved.

Is waist-to-height ratio a useful indicator of cardio-metabolic risk in 6-10-year-old children?

Background: Childhood obesity is a public health problem worldwide. Visceral obesity, particularly associated with cardio-metabolic risk, has been assessed by body mass index (BMI) and waist circumference, but both methods use sex-and age-specific percentile tables and are influenced by sexual maturity. Waist-to-height ratio (WHtR) is easier to obtain, does not involve tables and can be used to diagnose visceral obesity, even in normal-weight individuals. This study aims to compare the WHtR to the 2007 World Health Organization (WHO) reference for BMI in screening for the presence of cardio-metabolic and inflammatory risk factors in 6–10-year-old children. Methods: A cross-sectional study was undertaken with 175 subjects selected from the Reference Center for the Treatment of Children and Adolescents in Campos, Rio de Janeiro, Brazil. The subjects were classified according to the 2007 WHO standard as normal-weight (BMI z score > −1 and < 1) or overweight/obese (BMI z score ≥ 1). Systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting glycemia, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), Homeostatic Model Assessment – Insulin Resistance (HOMA-IR), leukocyte count and ultrasensitive C-reactive protein (CRP) were also analyzed. Results: There were significant correlations between WHtR and BMI z score (r = 0.88, p < 0.0001), SBP (r = 0.51, p < 0.0001), DBP (r = 0.49, p < 0.0001), LDL (r = 0.25, p < 0.0008, HDL (r = −0.28, p < 0.0002), TG (r = 0.26, p < 0.0006), HOMA-IR (r = 0.83, p < 0.0001) and CRP (r = 0.51, p < 0.0001). WHtR and BMI areas under the curve were similar for all the cardio-metabolic parameters. A WHtR cut-off value of > 0.47 was sensitive for screening insulin resistance and any one of the cardio-metabolic parameters. Conclusions: The WHtR was as sensitive as the 2007 WHO BMI in screening for metabolic risk factors in 6-10-year-old children. The public health message “keep your waist to less than half your height” can be effective in reducing cardio-metabolic risk because most of these risk factors are already present at a cut point of WHtR ≥ 0.5. However, as this is the first study to correlate the WHtR with inflammatory markers, we recommend further exploration of the use of WHtR in this age group and other population-based samples.

Untersuchungen zum Einfluss des HMG-CoA-Reduktase-Inhibitors Lovastatin auf die Toxizität ionisierender Strahlung sowie des Anthrazyklinderivats Doxorubicin im Mausmodell

Hintergund: HMG-CoA-Reduktase-Inhibitoren (Statine) sind klinisch etablierte Cholesterinsenker. Über die Inhibition der intrinsischen Cholesterinbiosynthese hinaus zeigen sie sogenannte pleiotrope biologische Effekte. Ein Großteil dieser Wirkungen wird auf die Inhibition kleiner Ras homologer GTPasen (Rho GTPasen) zurückgeführt. In vitro schützt das Statinderivat Lovastatin (Lova) primäre humane Endothelzellen vor der Zytotoxizität von ionisierender Strahlung (IR) und dem Krebsmedikament Doxorubicin (Doxo). Zielsetzung: Die Relevanz dieser Befunde für ein in vivo Mausmodell sollte in der vorliegenden Arbeit überprüft werden. Dafür wurden BALB/c-Mäuse mit IR oder Doxo behandelt und der Einfluss einer Kobehandlung mit Lova auf verschiedene Toxizitätsendpunkte untersucht (24 h nach einer einzelnen hohen Dosis IR (i), 14 Tage nach zwei geringen Dosen IR (ii), 48 h nach einer einzelnen hohen Dosis Doxo (iii), sowie 8 Tage nach drei niedrigen Dosen Doxo (iv)). Eine mögliche gleichzeitige Protektion von Tumorzellen durch die Statingabe wurde in einem Xenotransplantationsexperiment überprüft (v), in dem das gleiche Behandlungsschema wie bei iv angewendet wurde. Ergebnisse: Es konnte gezeigt werden, dass eine Statinbehandlung Normalgewebe vor Doxo- und IR-induzierter Toxizität schützt, ohne gleichzeitig protektiv auf transformierte Zellen zu wirken. Dieser Effekt ist wahrscheinlich von einer Inhibition der kleinen GTPasen Rac1 und RhoA abhängig und einer daraus folgenden Modifizierung der DNA-Schadensantwort. i: Die Statinvorbehandlung der Mäuse hatte keinen Einfluss auf die Bildung von initialen IR-induzierten DNA-Doppelstrangbrüchen (DSB) in der Leber. Die Lova-Behandlung wirkte sich jedoch auf IR-induzierte Stressantworten aus, was sich in einer Minderung der Expression von Inflammations- und Fibrosesurrogatmarkern in Leber und Darm widerspiegelte. ii: In der Lunge der Tiere wurde ein Anstieg von molekularen Inflammations- und Fibrosesurrogatmarkern detektiert, der bei Statinkobehandlung ausblieb. Zudem verhinderte die Kobehandlung mit Lova eine IR-induzierte Abnahme der Thrombozytenzahl, ohne sich auf die durch IR verringerte Leukozytenzahl im Blut auszuwirken. iii: Die Verabreichung einer hohen Dosis Doxo induzierte DSB-Formation in der Leber. Die Statinvorbehandlung reduzierte deren Menge um ca. 50 %. Dieser genoprotektive Effekt war unabhängig von der Entstehung reaktiver Sauerstoffspezies sowie einer Änderung des Doxo-Imports oder Exports. Die Expression von proinflammatorischen und profibrotischen Genen fiel besonders in der Leber und im Herzen durch die Lova-Kobehandlung geringer aus, als in der nur mit Doxo behandelten Gruppe. Zudem verringerte Lova die durch Doxo induzierte Hochregulation von für den AP1-Komplex kodierenden Genen sowie von Zellzykluskontrollfaktoren. Die Lova-Vorbehandlung führte darüber hinaus im Herzen zu einem reduzierten mRNA-Spiegel der Topoisomerasen II α und β. iv: Es konnten schwere Herz- und Leberschäden detektiert werden (gemessen an Gldh-, Gpt- sowie cTn-I-Serumkonzentrationen), die bei einer Kobehandlung mit dem Statin nicht auftraten. Die Lova-Kobehandlung verhinderte außerdem eine durch die Doxo-Behandlung verringerte Leukozytenzahl. Molekulare Marker für frühe fibrotische Ereignisse, sowie für Inflammation und Hypertrophie waren in der Leber und im Herzen nach der Doxo-Behandlung erhöht. Das Statin war auch hier in der Lage, diese toxischen Wirkungen des Anthrazyklins zu mindern. Auch die Doxo-induzierte Expression von Surrogatmarkern für Zellantworten auf oxidativen Stress wurde in der Leber abgeschwächt. In der Leber und im Herzen wiesen die mit Doxo behandelten Tiere höhere mRNA Spiegel von an Zellzykluskontrolle beteiligten Faktoren sowie von DNA-Reparatur und Fremdstoffmetabolismus assoziierten Genen auf. Am stärksten wurde die Expression von Topoisomerase II alpha - ein molekularer Marker für Zellproliferation und bedeutsame Zielstruktur von Doxo - in der Leber hochreguliert. Die Statin-Kobehandlung verhinderte all diese Doxo-induzierten Expressionsänderungen. Im Gegensatz zur Leber wurde die Top2a-mRNA Menge im Herzen durch die Doxo-Applikation reduziert. Auch hier bewirkte die Kobehandlung mit dem Statin, dass die Expression nahe dem Kontrollniveau blieb. v: Die Kobehandlung mit Lova führte zu keinem Schutz der Tumorzellen vor Doxo, sondern erhöhte sogar dessen antineoplastisches Potential.rnFazit: Die Erkenntnisse aus vorhergegangenen in vitro Versuchen konnten zum großen Teil auf die in vivo Situation im Mausmodell übertragen werden. Sie stehen im Einklang mit Ergebnissen anderer Gruppen, welche die Inhibition kleiner GTPasen mit einer geringeren, durch zytotoxische Substanzen induzierten, Inflammation und Fibrose korrelieren konnten. Eine Kobehandlung mit Lova während einer Krebstherapie erscheint somit als vielversprechende Möglichkeit Doxo- oder IR-induzierte Nebenwirkungen auf Normalgewebe zu mildern.

Predicting adverse events in children with fever and chemotherapy-induced neutropenia: the prospective multicenter SPOG 2003 FN study

PURPOSE To develop a score predicting the risk of adverse events (AEs) in pediatric patients with cancer who experience fever and neutropenia (FN) and to evaluate its performance. PATIENTS AND METHODS Pediatric patients with cancer presenting with FN induced by nonmyeloablative chemotherapy were observed in a prospective multicenter study. A score predicting the risk of future AEs (ie, serious medical complication, microbiologically defined infection, radiologically confirmed pneumonia) was developed from a multivariate mixed logistic regression model. Its cross-validated predictive performance was compared with that of published risk prediction rules. Results An AE was reported in 122 (29%) of 423 FN episodes. In 57 episodes (13%), the first AE was known only after reassessment after 8 to 24 hours of inpatient management. Predicting AE at reassessment was better than prediction at presentation with FN. A differential leukocyte count did not increase the predictive performance. The score predicting future AE in 358 episodes without known AE at reassessment used the following four variables: preceding chemotherapy more intensive than acute lymphoblastic leukemia maintenance (weight = 4), hemoglobin > or = 90 g/L (weight = 5), leukocyte count less than 0.3 G/L (weight = 3), and platelet count less than 50 G/L (weight = 3). A score (sum of weights) > or = 9 predicted future AEs. The cross-validated performance of this score exceeded the performance of published risk prediction rules. At an overall sensitivity of 92%, 35% of the episodes were classified as low risk, with a specificity of 45% and a negative predictive value of 93%. CONCLUSION This score, based on four routinely accessible characteristics, accurately identifies pediatric patients with cancer with FN at risk for AEs after reassessment.

Clinical Characteristics of Children with Viral Single- and Co-Infections and a Petechial Rash

BACKGROUND:: Children with petechial rash are more likely to undergo invasive diagnostics, to be treated with antibiotics for potential bacterial infection and to be hospitalized. However, viruses have also been associated with petechial rash. Nonetheless, a systematic analysis of viral infections with modern available techniques as quantitative real time polymerase chain reaction (q-PCR) in the context of petechial rash is lacking. The purpose of this pediatric study was to prospectively uncover viral pathogens that may promote the emergence of petechiae and to analyze the correlation with the clinical characteristics and course. METHODS:: We conducted a prospective study in children (0 to 18 years) presenting with petechiae and signs or symptoms of infection at the emergency department between November 2009 and March 2012. In nasopharyngeal aspirates the following viruses were analyzed by q-PCR: Cytomegalovirus, Epstein-Barr virus, parvovirus B19, Influenza A and B, parainfluenza viruses, human respiratory syncytial virus A and B, human metapneumovirus, rhinovirus, enterovirus, adenovirus, human coronavirus OC43, 229E, NL63 and human bocavirus. RESULTS:: A viral pathogen was identified in 67% of the analyzed 58 cases with petechial rash. Virus positive patients showed a significantly higher incidence of lower respiratory tract infections. Forty-one percent were viral co-infections, which were significantly younger than virus negative patients, had a higher leukocyte count and were hospitalized for a longer time. CONCLUSIONS:: A petechial rash is frequently associated viral single- and co-infections and can rapidly be identified via q-PCR.

Successful lower extremity angioplasty improves brachial artery flow-mediated dilation in patients with peripheral arterial disease

INTRODUCTION: Peripheral arterial disease (PAD) is associated with systemic impaired flow-mediated dilation (FMD) and increased risk for cardiovascular events. Decreased FMD may be caused by a decrease in arterial shear stress due to claudication and inflammation due to muscle ischemia and reperfusion. We assumed that endovascular revascularization of lower limb arterial obstructions ameliorates FMD and lowers inflammation through improvement of peripheral perfusion. METHODS: The study was a prospective, open, randomized, controlled, single-center follow-up evaluation assessing the effect of endovascular revascularization on brachial artery reactivity (FMD) measured by ultrasound, white blood cell (WBC) count, high-sensitive C-reactive protein (hs-CRP), and fibrinogen. We investigated 33 patients (23 men) with chronic and stable PAD (Rutherford 2 to 3) due to femoropopliteal obstruction. Variables were assessed at baseline and after 4 weeks in 17 patients (group A) who underwent endovascular revascularization and best medical treatment, and in 16 patients (group B) who received best medical treatment only. RESULTS: FMD did not differ between group A and B (4.96% +/- 1.86% vs 4.60% +/- 2.95%; P = .87) at baseline. It significantly improved after revascularization in group A (6.44% +/- 2.88%; P = .02) compared with group B at 4 weeks of follow-up (4.53% +/- 3.17%; P = .92), where it remained unchanged. The baseline ankle-brachial index (ABI) was similar for group A and B (0.63 +/- 0.15 vs 0.66 +/- 0.10; P = .36). At 4 weeks of follow-up, ABI was significantly increased in group A (1.05 +/- 0.15; P = .0004) but remained unchanged in group B (0.62 +/- 0.1). WBC counts of the two groups were comparable at baseline (group A: 7.6 +/- 2.26 x 10(6)/mL and group B: 7.8 +/- 2.02 x 10(6)/mL, P = .81). In group A, the leukocyte count significantly decreased after angioplasty from 7.6 +/- 2.26 to 6.89 +/- 1.35 x 10(6)/mL (P = .03). For group B, WBC count did not differ significantly compared with baseline (7.76 +/- 2.64 x 10(6)/mL; P = .94). No effects were observed on hs-CRP or fibrinogen from endovascular therapy. CONCLUSION: Endovascular revascularization with reestablishment of peripheral arterial perfusion improves FMD and reduces WBC count in patients with claudication. Revascularization may therefore have clinical implications beyond relief of symptoms, for example, reducing oxidative stress caused by repeated muscle ischemia or increased shear stress due to improved ambulatory activity.