Ixolaris, a tissue factor inhibitor, blocks primary tumor growth and angiogenesis in a glioblastoma model

Background: The expression levels of the clotting initiator protein Tissue Factor (TF) correlate with vessel density and the histological malignancy grade of glioma patients. Increased procoagulant tonus in high grade tumors (glioblastomas) also indicates a potential role for TF in progression of this disease, and suggests that anticoagulants could be used as adjuvants for its treatment. Objectives: We hypothesized that blocking of TF activity with the tick anticoagulant Ixolaris might interfere with glioblastoma progression. Methods and results: TF was identified in U87-MG cells by flow-cytometric and functional assays (extrinsic tenase). In addition, flow-cytometric analysis demonstrated the exposure of phosphatidylserine in the surface of U87-MG cells, which supported the assembly of intrinsic tenase (FIXa/FVIIIa/FX) and prothrombinase (FVa/FXa/prothrombin) complexes, accounting for the production of FXa and thrombin, respectively. Ixolaris effectively blocked the in vitro TF-dependent procoagulant activity of the U87-MG human glioblastoma cell line and attenuated multimolecular coagulation complexes assembly. Notably, Ixolaris inhibited the in vivo tumorigenic potential of U87-MG cells in nude mice, without observable bleeding. This inhibitory effect of Ixolaris on tumor growth was associated with downregulation of VEGF and reduced tumor vascularization. Conclusion: Our results suggest that Ixolaris might be a promising agent for anti-tumor therapy in humans.

Labile Plasma Iron Generation After Intravenous Iron is Time-dependent and Transitory in Patients Undergoing Chronic Hemodialysis

Iron supplementation in hemodialysis patients is fundamental to erythropoiesis, but may cause harmful effects. We measured oxidative stress using labile plasma iron (LPI) after parenteral iron replacement in chronic hemodialysis patients. Intravenous iron saccharate (100 mg) was administered in patients undergoing chronic hemodialysis (N = 20). LPI was measured by an oxidant-sensitive fluorescent probe at the beginning of dialysis session (T0), at 10 min (T1), 20 min (T2), and 30 min (T3) after the infusion of iron and at the subsequent session; P < 0.05 was significant. The LPI values were significantly raised according to the time of administration and were transitory: -0.02 +/- 0.20 mu mol/L at the beginning of the first session, 0.01 +/- 0.26 mu mol/L at T0, 0.03 +/- 0.23 mu mol/L at T1, 0.09 +/- 0.28 mmol/L at T2, 0.18 +/- 0.52 mmol/L at T3, and -0.02 +/- 0.16 mmol/L (P = 0.001 to 0.041) at the beginning of the second session. The LPI level in patients without iron supplementation was -0.06 +/- 0.16 mmol/L. Correlations of LPI according to time were T1, T2, and T3 vs. serum iron (P = 0.01, P = 0.007, and P = 0.0025, respectively), and T2 and T3 vs. transferrin saturation (P = 0.001 and P = 0.0003, respectively). LPI generation after intravenous saccharate administration is time-dependent and transitorily detected during hemodialysis. The LPI increment had a positive correlation to iron and transferrin saturation.

A PPCCIPS: a Política Pública da Coleta Complementar Itinerante e Programada de Sangue, sob a luz da teoria do ciclo da política pública

This study aimed to focus on aspects of public administration concerning the implementation of the public policy of complementary blood collection by the itinerant and scheduled PPCCIPS services either trough off local unity or mobile unit blood collection operations, which are managed by the State Institute of Hematology Arthur de Siqueira Cavalcanti - HEMORIO. The case study method was used in that public health institutional field, in search for a better understanding of responsibilities and management related to collection, serology, fractionation, storage and distribution of blood supply to almost all public hospitals and clinics, summed up to agreements with the single health system of the State of Rio de Janeiro. Bibliographic references, documentary and field data obtained through interviews and systematic observation in the public servants of HEMORIO workplaces, were treated by the analysis of the content method and the results of this research revealed the complexity of those services, and needs in outstanding aspects of infrastructure, equipment, logistics and personnel, which are critical for the achievement of the increased public collection of blood in the Rio de Janeiro State, endorsing the suggestions for the implementation of PPCCIPS in HEMORIO. The main point found in this research results concern the immanent ethical commitment of that public service personnel, including staff members and low ranking members as well, perceived due to a brief philosophical overlook on that personnel¿s attitudes. An important strategic aspect was revealed by the need for excellence of midia communications and education programs to implement the community involvement in the whole process. Final reflections point out that personnel posture is considered vital for the quality of the expected care of the technical activities and also for the quality of its final products release to the local public, fluminense, which is the irreplaceable human blood, and their derivatives. Despite the author¿s effort in this dissertation there is much more to be studied on that crucial theme.

Hematological analysis of Micropogonias Furnieri, Desmarest, 1823, Scianidae, from two estuaries of Baixada Santista, São paulo Brazil

Alterações hematológicas em peixes são consideradas uma importante ferramenta para avaliar processos patológicos decorrentes da exposição a poluentes ambientais. Micropogonias furnieri (Desmarest, 1823) (corvina) é comumente encontrada em regiões estuarinas e eventualmente está exposta a inúmeros contaminantes. No presente estudo foi avaliado o quadro hematológico de indivíduos de M. furnieri coletados na Baixada Santista: o Sistema Estuarino de Santos, considerado poluído, e o estuário do Rio Itanhaém (controle). Foram avaliados o número de Eritrócitos (Er), o Hematócrito (Ht), a taxa de Hemoglobina (Hb), o Volume Corpuscular Médio (VCM) e a Concentração de Hemoglobina Corpuscular Média (CHCM). Nos peixes coletados no Sistema Estuarino de Santos, os níveis de Ht foram significativamente menores, enquanto os níveis de CHCM e Hb foram significativamente mais altos, indicando que os prováveis efeitos estejam atribuídos aos diferentes níveis de contaminação encontrados nos estuários.

Efeito de um extrato de Ganoderma lucidum (Reishi) na marcação de constituintes sanguíneos com tecnécio-99M e na sobrevivência de Escherichia coli

Clinical evaluations have been made possible with radiobiocomplexes marked with tecnecium-99m (99mTc). Natural or synthetic drugs are able to interfere in the marking of blood structures with 99m Tc. Also, the toxicity of several natural products has been described. The aim of this study was evaluating the effect of an extract of Ganoderma lucidum (Reishi) in the marking of blood constituents with 98m Tc and in the survival of Escherichia coli. Blood samples from Wistar rats were treated with reishi extract. Radiomarking procedure was performed. Samples of plasma (P), blood cells (CS), and insoluble (FI) and soluble (FS) fractions of P and CS were separated and the radioactivity was counted to determine radioactivity percentages (%ATI). Escherichia coli AB1157 cultures were treated with stannous chloride in the presence and absence of the reishi extract. Blood samples and bacterial cultures treated with NaCl 0.9% were used as controls. Data indicated that the reishi extract has significantly altered (p<0,05) the %ATI of P, CS, FI-P, FS-P, FI-CS e FS-CS, as well as it has increased survival of bacterial cultures treated with stannous chloride. Our results suggest that the Reishi extract would be able to present a redox/ chelant action by altering blood constituent marking with 99mTc and by protecting bacterial cultures against stannous chloride-induced oxydating lesions. The study had a multidisciplinary character, with the participation of the following areas of knowledge: Biophysics, Radiobiology, Botanics, Phytotherapy, and Hematology

Metacromasia para identificação de basófilos sanguíneos em surubim híbrido: contribuição metodológica

Padua S.B. & Ishikawa M.M. [Metachromasia for blood basophils identification in hybrid surubim catfish: methodological contribution]. Metacromasia para identificacao de basofilos sanguineos em surubim hibrido: contribuicao metodologica. Revista Brasileira de Medicina Veterinaria, 33(3):147-150, 2011. Centro de Aquicultura da Unesp/Jaboticabal Rod. Paulo Donato Castellane, s/n Bairro Rural, Jaboticabal, 14884-900, SP. Brasil. E-mail: santiagopadua@live.comDifferent protocols of fixation and hydrolysis of blood smears of hybrid surubim catfish for metachromasia to marking blood basophils were evaluated. For this reason, methanol, acid-alcohol and formalin vapor were tested as fixatives. For hydrolysis of blood smears, HCl, citric acid and 2-Mercaptoethanol + Urea + NaCl solutions was evaluated. After procedures, the blood smears was stained with toluidine blue (0.025%) diluted in McIlvaine buffer (pH 4). The different protocols for fixation and hydrolysis of blood smears for metachromasia influenced the quality of the reaction. Hydrolysis with 2-Mercaptoethanol + Urea + NaCl solutions provided negative results when using methanol and acid-alcohol as fixatives. The fixation with acid-alcohol associated with citric acid hydrolysis provides highest quality reason.

HEMATOLOGICAL ASSESSMENT IN FOUR BRAZILIAN TELEOST FISH WITH PARASITIC INFECTIONS, COLLECTED IN FEEFISHING FROM FRANCA, São Paulo, BRAZIL

The effects of parasitic infections in condition factor, hematocrit, hemoglobin, mean corpuscular hemoglobin concentration (MCHC), and leucocytes and thrombocytes distribution in Piaractus mesopotamicus, Leporinus macrocephalus, hybrid tambacu (P. mesopotamicus x Colossoma macropomum and Brycon amazonicus collected in feefishing from Franca, São Paulo, Brazil were evaluated. Parasitized tambacu and L. macrocephalus had higher (p<0.05) condition factor than unparasitized fish. However, the contrary occurred in P. mesopotamicus and B. amazonicus. Changes in the hematocrit, hemoglobin and MCHC were not related to parasitism. Parasitic infections did not cause effect on leucocytes and thrombocytes percentage (p>0.05) of tambacu. In P. mesopotamicus parasitized by Monogenea Anacanthorus penilabiatus and dinoflagellate Piscinoodinium pillulare, increase in monocytes and decrease in thrombocytes percentage (p<0.05) were found. However, the same parasitic association in L. macrocephalus caused a decrease in lymphocytes percentage accompanied by increase in neutrophils percentage (p<0.05). In B. amazonicus, infection by Ichthyophthirius multifiliis, P. pillulare and monogeneans caused increase in neutrophils percentage.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

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

Tempo de migração dos macrófagos em matrinxã, Brycon amazonicus, por meio da técnica de inoculação de leveduras Saccharomyces cerevisiae

Na aquicultura são utilizados análises da ativação e incremento da migração de macrófagos, com intuito de verificar a capacidade imunológica inespecífica dos peixes frente a um desafio. Neste sentido, o objetivo deste estudo foi determinar o tempo de migração de monócitos/macrófagos para a cavidade peritoneal em matrinxã, Brycon amazonicus, por meio da técnica de inoculação de leveduras Saccharomyces cerevisiae, e verificar as possíveis alterações dos parâmetros hematológicos após o estímulo. Foram utilizados 30 matrinxãs com peso médio de 101,55 ± 24,50 g e comprimento médio de 19,75 ± 1,72 cm. Os tempos de inoculação utilizados foram 2, 4, 8 e 12 horas, sendo utilizados 6 animais por tempo. Após os períodos de incubação (2, 4, 8 e 12 horas), os exemplares foram anestesiados e alíquotas de sangue foram coletadas por punção do vaso caudal, para a análise: número total de células, contagem diferencial e total dos leucócitos e contagem total de trombócitos, hematócrito, taxa de hemoglobina e índices hematimétricos (VCM, HCM e CHCM). Os resultados mostram que a capacidade fagocítica do macrófago não apresentou diferenças significativas entre os tempos experimentais. Com relação ao índice fagocítico, o tempo de 2 horas representa o tempo em que os macrófagos fagocitaram maior número de leveduras com diferenças significativas em relação aos outros tempos experimentais, indicando que este tempo (2 horas) de incubação foi suficiente para a migração e ativação máxima dos macrófagos da cavidade peritoneal, da espécie estudada. Os valores do número de eritrócitos apresentaram diferenças entre os tempos de incubação. Entretanto, os valores dos outros parâmetros hematológicos não apresentaram diferenças significativas.

Influência de diferentes níveis de vitamina C na ração sobre parâmetros hematológicos de alevinos de Piaractus mesopotamicus Holmberg (Osteichthyes, Characidae)

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

Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta

Beta thalassemia arises as a consequence of the reduction (β+, β++, βsilent) or absence (β0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) and codon 39 (CAG®TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our country

Avaliação dos marcadores celulares por citometria de fluxo nos portadores de leucemia mieloide aguda atendidos no Hemocentro do Rio Grande do Norte-Hemonorte

The acute myeloid leukemia (AML) is a disease in which malignant myeloblasts expand, build up and suppress normal hematopoietic activity would represent a major diagnostic challenge. With the advent of immunophenotyping by flow cytometry, the diagnosis of these tumors have become more faithful, facilitating the treatment and monitoring of patients. The objectives of this study: diagnosis and classification of AML based on immunophenotyping by flow cytometry with a panel of AcMo specific for acute leukemias, set the frequency of AML in samples from patients with acute leukemias sent to the Department of Hematology Blood Center of Rio Grande do Norte - HEMONORTE, establish standards of antigen expression for different subtypes of acute leukemia and its correlation with the newly diagnosed cases refractory to treatment and recurrence of the disease, standardization of methods for detection and labeling of surface antigens by flow cytometry and intracytoplasmic flow, and observe the frequency of acute leukemia with aberrant phenotypes rare. During the study, 351 were diagnosed acute leukemia, and 179 (51%) classified as AML and 172 (49%) and ALL, which were excluded from the present work. Of the 179 AML, 92 (51.4%) were female and 87 (48.6%) were male, with ages ranging from 3 to 95 years of ag, with higher incidence in individuals in the age group of 41 to 65. Splenomegaly was the clinical finding more present, a total of 147 cases (82.1%), followed by hepatomegaly present in 132 cases (73.7%). The hemorrhagic events were observed in 55 cases (30.7%). Lymphadenopathy in turn was detected in 20 of 179 cases (11.2%). In order to classify subtypes of AML, we used a large panel of monoclonal antibodies, obtaining the following results: AML M0, 02 (1.1%) AML M1, 40 (22.3) AML M2, 60 (33.5) AML M3, 22 (12.3%) AML M4, 10 (5.6) AML M5, 13 (7.3%) AML M6 06 (3.4%) and AML M7 01 (0.6%). We observed some cases with aberrant expression of some antigens such as CD7, CD4, CD19, CD3, CD5 and TdT, CD 7 was present in 30 (16.8%), CD4 in 5 (2.8%), the CD 3 in 5 (2.8%), the CD19 in 3 (1.7%), the CD5 in 3 (1.7%) and TDT was in 7 (3.9%) cases of AML .the CD8 and CD79a was present in only a 1 case.

Effects of physical and chemical immobilization on hematologic and biochemical variables in captive brown brocket deer (Mazama gouazoubira)

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

Respostas hematológicas em tuvira após anestesia com diferentes concentrações de óleo de cravo

The hematological responses in tuvira (Gymnotus aff. inaequilabiatus) after clove oil anesthesia at different concentrations were evaluated in this study. Fish (n = 25) with average length of 26.8 +/- 2.4 cm and weighing 94.4 +/- 23.6 g were separated in five groups composed with five animals each and submitted individually to respective concentrations of clove oil: 40, 80, 120, 160 and 200 mg L-1. After anesthetic recuperation, the blood samples collection were performed, as well as complete hemogram. The results were submitted to analysis of variance, Tukey test (P<0.05) and Sperman's correlation analysis. There was an increase (P<0.05) on hematocrit, red blood cells count, thrombocytes count and in the values of neutrophils with increasing of clove oil concentrations. In addition, positive correlation (P<0.05) were observed on total leukocytes, as well as negative linear correlation (P<0.05) for mean corpuscular hemoglobin concentration (MCHC). The clove oil anesthesia on tuvira cause hematological changes with an effect dose-dependent. These changes should be considered mainly in future studies designed to establish the reference values of hematologic variables for this specie.

Estudo eritroleucométrico e proteinograma sérico do sangue do cordão umbilical e jugular de eqüinos ao nascimento e de suas respectivas mães

Colheram-se amostras de sangue do cordão umbilical (SCU) e do sangue circulante de cinco eqüinos neonatos, imediatamente após o nascimento, e o sangue da própria mãe, utilizando-se um sistema a vácuo. O material foi submetido à contagem global de hemácias e leucócitos e à determinação do volume globular e da concentração de hemoglobina; à contagem diferencial de leucócitos em esfregaços sangüíneos; e ao cálculo dos índices eritrocitométricos. Foram realizadas a dosagem de proteínas séricas totais e a eletroforese das proteínas séricas em gel de agarose. Não houve diferenças significativas entre os parâmetros do SCU e do sangue da jugular dos potros. No SCU dos potros observaram-se valores mais elevados para contagem global de hemácias (9,75x10(6)/µl), dosagem de hemoglobina (14,65g/dl) e concentração de hemoglobina corpuscular média (37,23g/dl); e valores menores para volume corpuscular médio (40,50fl), proteína total (4,37g/dl), alfa-globulinas (0,65g/dl), beta-globulinas (1,10g/dl), gama-globulinas (0g/dl) e contagens global (5,40 x 10³/µl) e diferencial de leucócitos, exceto contagem de neutrófilos bastonetes e monócitos, quando comparados com os valores obtidos no sangue de suas mães.