Prevalence of thrombophilia-associated genetic risk factors in blood donors of a regional hospital in southern Brazil

ABSTRACT Introduction: Thromboembolic events occur due to an imbalance in the hemostasis and some factors associated with this condition can be inherited. In order to evaluate the frequency of genotypes considered to be common hereditary risk factors for thrombophilia associated with venous thrombosis (g.1691G>A and g.20210G>A) and hyperhomocysteinemia (g.677C>T and g.1298A>C), samples from voluntary healthy blood donors at the Hospital de Clínicas de Porto Alegre were tested. Methods: We examined 325 blood samples from blood donors collected from October 2017 to July 2018. Blood was collected on filter paper and the DNA was extracted for single nucleotide polymorphisms (SNPs) analysis using the qualitative real time polymerase chain reaction. Results: The calculated frequencies of each genetic variant in heterozygosity were 4% for the FV gene (g.1691G> A), 4% for the F2 gene (g.20210G> A) and 42% and 39% for methylenetetrahydrofolate reductase (MTHFR), g.677C>T and g.1298A>C, respectively. Only the genetic variants of MTHFR were found in homozygosity, with frequencies of 14% and 6% (g.677C>T and g.1298A>C), respectively. Discussion: Altogether, these results describe the frequencies of genetic variants associated with venous thrombosis and hyperhomocysteinemia in the analyzed group and are important to enhance our current knowledge about the genetic profiles of Brazilian blood donors.

Quantification of peripheral blood CD34+ cells prior to stem cell harvesting by leukapheresis: a single center experience

ABSTRACT Background: Due to laboratory logistic issues, our center has traditionally scheduled peripheral blood stem cell harvests based on timing from the start of mobilization. This has proved to be useful in some cases, but also resulted in many fruitless harvests due to poor mobilization. In order to improve the efficiency of collections and compare the effectiveness of peripheral blood CD34+ cells as a predictor with data from other reports, this study analyzed the implementation of this routine. Methods: Peripheral blood and leukapheresis samples were quantified by flow cytometry and the association between these parameters was assessed. Results: Sixty-six consecutive leukapheresis samples were collected from 34 patients after the collection of peripheral blood samples for CD34+ quantification. A moderate positive correlation was observed between peripheral blood CD34+ cell count and total CD34+ cell count/kg (r = 0.596; p-value < 0.001). A multivariable regression model also confirmed this association and allowed the estimation that for every increase in five CD34+ cells/µL in the peripheral blood, a mean increase of 0.38 × 106 CD34+ cells/kg could be predicted. Demographic characteristics, baseline comorbidities and mobilization regimen did not influence final CD34+ cell count in this sample. Conclusions: As observed in other centers, quantification of peripheral blood CD34+ progenitor cells is a strong predictor of effectiveness to guide stem cell harvesting. Due to the results of this study, a modification in the peripheral blood stem cell harvesting logistics was implemented at our center in order to incorporate this routine.

Blood transfusion-related immunomodulation

The phenomenon of transfusion-related immunomodulation (TRIM) has been studied since the observation of a higher kidney allograft survival in patients who had received a higher number of transfusions. Conversely, it has been suggested as one of the possible causes related to the development of infections in patients with multiple blood transfusions and/or after a major surgery, and has been also associated with a decreased function of natural killer cells (NK) and antigen-presenting cells (APCs), reduced cell-mediated immunity, and increased regulatory T cells (Tregs). This review aimed to conceptualize TRIM and discuss some aspects related to its mechanisms and the prevention of immunomodulatory events.

Trombocitose essencial: o que é essencial saber / Essential thrombocytosis: what is vital to know

A trombocitose essencial (TE) faz parte do grupo de síndromes mieloproliferativas (SMP) cromossomo Philadelphia(Ph) negativas. Caracteriza-se pela hiperproliferação megacariocítica com consequente trombocitose periférica, favorecendo fenômenos trombo-hemorrágicos. Esta entidade estava esquecida até meados de 2005, quando as recentes publicações sobre as alterações moleculares na atividade da enzima tirosina quinase, JAK2, desencadeou um novo interesse sobre a patogenia, aspectos clínicos e terapêuticos da TE. A identificação das mutações de JAK2 e do gene MPL W515K, W515L e S505N impulsionou a nova proposta da Organização Mundial de Saúde (OMS) para reformular os critérios diagnósticos, reduzindo o número de plaquetas para 450x10(9)/L. O alicerce do tratamento são agentes redutores das contagens plaquetárias: hidroxiureia, anagrelide ou interferon associados à prevenção das complicações trombo-hemorrágicas. Não há um tratamento curativo para a TE, mas despontam perspectivas de que terapias alvo, bloqueadoras da mutação JAK2, possam incrementar o desfecho da doença. Inibidores de JAK2, específicos e inespecíficos, estão sendo estudados em fase I e II e parecem promissores num futuro próximo.

Essential thrombocythemia (ET) is an acquired myeloproliferative Philadelphia negative disorder characterized by megakaryocytic hyperproliferation and persistent peripheral thrombocytosis with a tendency of thrombosis and hemorrhages. This entity was forgotten until 2005, when the recent identification of somatic mutations such as JAK2V617F and MPL W515L/K triggered off interest in the molecular pathogenesis, clinical aspects and therapeutic approach of ET. The presence of molecular mutations changed the diagnostic criteria proposed by the World Health Organization, and nowadays the platelet count for which ET should be considered has dropped to 450 X 10(9) /L. Treatment is given according to risk stratification: in cases with high risk platelet reduction, therapy using drugs such as hydroxyurea, interferon or anagrelide is chosen. There is no drug known to cure ET and the current therapy is either to prevent thrombohemorrhagic events or reductions in the platelet count. The identification of the JAK2V617F mutation has opened an opportunity to develop new therapeutic target. JAK2 inhibitors are promising for the treatment of ET in the near future.

Talidomida e mieloma múltiplo: verificação dos efeitos terapêuticos através de parâmetros clínico e laboratoriais / Thalidomide and multiple myeloma: therapy evaluation using clinical and laboratorial parameters

Nas duas últimas décadas, houve uma mudança radical na terapia e na evolução do mieloma múltiplo(MM), neoplasia hematológica ainda considerada fatal. As pesquisas e investimentos em medicamentos que interferem com a fisiopatogenia e com o microambiente medular estão permitindo o controle e a regressão do clone plasmocitário maligno, mudando as perspectivas da doença. A idéia nova de usar uma droga velha, a talidomida, tem-se mostrado efetiva no MM. Em 1997, apostando nos efeitos imunomoduladores e antiangiogênicos da talidomida, foram iniciados ensaios clínicos para MM refratários. A partir daí, outras ações sobre o plasmócito e microambiente medular foram eficazes contra a doença, não somente em refratários ou recaídos, mas também como terapia de indução e/ou de manutenção da remissão. No Serviço de Hematologia do Hospital de Clínicas de Porto Alegre foram acompanhados 35 portadores de mieloma múltiplo, em uso de doses baixas (100 mg) de talidomida, pelas indicações: 13 - manutenção pós-TMO, 11 - pós-indução, 5 - recaída, 4 - refratariedade e 2 - terapia de indução. O estudo vigorou entre março/01 a dez/03. Os parâmetros avaliados foram: nível Hb, pico da imunoglobulina sérica ou urinária e o número de plasmócitos na medula óssea. As medidas foram tomadas pré-talidomida e após 3, 6 e 12 meses. A taxa de imunoglobulina foi o padrão ouro para avaliação de resposta. Os resultados: a dose terapêutica tolerada em 48 por cento dos pacientes foi 100 mg; 65 por cento dos tratados para induzir remissão (11 pacientes) apresentaram melhora entre 25 por cento-50 por cento no nível da imunoglobulina sérica; 87,5 por cento daqueles que usaram para manutenção de remissão (13 pós-TMO/ 11 pós-indução) mantiveram o mesmo plateau inicial.

Over the last two decades, we have seen a radical change intherapy and progression of multiple myeloma, a malignanthematologic disease that is still considered fatal. Recentinvestment and research on mechanisms that interfere in thephysiopathogenesis and bone marrow microenvironment areturning control and regression of the malignant plasma cellclone into something achievable, which may change expectationsrelated to this disease. The new idea of using an old drug,thalidomide, has shown to be effective in multiple myeloma. In1997, using the known effects of immunomodulation and antiangiogenesisof this drug, clinical trials were started in patients with unresponsive disease. Other therapeutic interventions inthe bone marrow microenvironment and plasma cells have beenadded and proved to be efficacious, not only as a therapy forrefractory patients, but also for induction and/or remissionmaintenance therapy. Thirty-five patients with multiple myelomawere treated with low-dose thalidomide (100 mg) and followedup. .... The study tookplace in the Hematology and Bone Marrow Transplantationservice of the Hospital de Clínicas de Porto Alegre, from March2001 to December 2003. Hemoglobin levels, serum or urineimmunoglobulin peaks and bone marrow plasma cell countswere evaluated. These parameters were assessed before startingwith the drug and after 3.6 and 12 months of usage. Theimmunoglobulin level was considered the gold standard toevaluate the response. The results showed that 100 mg was thetolerable dose for 51% of the patients. Sixty-five percent of thosewho used thalidomide for induction therapy showed a 25 to 50%improvement in immunoglobulin serum levels and 90% of thepatients on maintenance therapy (13 after bone marrowtransplantation, 11 after induction), sustained the sameimmunoglobulin levels of the initial plateau.

Leucemia Mielóide Agura: perfil de duas décadas do Serviço de Hematologia do Hospital das Clínicas de Porto Alegre - RS / Acute Myelogenous Leukemia: two decades overview - Hematology Service Hospital de Clínicas de Porto Alegre - RS

Nos últimos vinte anos, nosso serviço recebeu os pacientes com a suspeita de Leucemia Mielóide Aguda (LMA) provenientes de todas as regiões do estado. Entre março de 1980 e dezembro de 1999 analisamos 195 pacientes com idades superior a 12 e inferior a 70 anos, apresentando LMA "de novo" excetuando o subtipo M3. Na década de 80 foram registrados 102 pacientes: 47 homens e 55 mulheres. Destes, 84 receberam quimioterapia de indução com Citarabina e Daunorrubicina (esquema "7+3"), resultando no índice de remissão de 51 por cento (43/84). As médias de sobrevidas livre de doença e global foram dez meses em 35 por cento e 12 meses em 13 por cento respectivamente. De jan/90 a dez/93 houve 41 novos diagnósticos e todos foram submetidos à quimioterapia com esquema "7+3" atingindo a taxa de 66 por cento de remissão. As sobrevidas livre de doença e global foram estatisticamente (p<0.001) superiores com 17,6 meses em 41 por cento e 22 meses em 17,5 por cento dos pacientes. Em 1994 foi instituído novo protocolo orientado pelo grupo cooperativo brasileiro de estudos de leucemia (CBEL), preconizando a indução com Citarabina +Idarrubicina, seguido dos ciclos de consolidação e dois ciclos de intensificação com altas doses. Entre 1994-99 foram incluídos 52 pacientes neste protocolo, cujo índice de remissão foi 73 por cento (p=0,002). Em 50 por cento destes pacientes, a sobrevida livre de doença alcançou 23,3 meses, enquanto a sobrevida global foi de 26 meses em 25 por cento. As sobrevidas livre de doença e global, neste último período, tiveram diferenças estatisticamente significativas comparadas aos anos anteriores. Observamos uma melhora nas taxas de remissão e de sobrevida na última década, mesmo assim a sobrevida longa livre de eventos permanece inferior a 30 por cento. Nossos resultados são equivalentes aos demais centros nacionais que tratam LMA e corroboram os dados da literatura

We have treated many AML patients in the last twenty years. We followed, between March 1980 andDecember 1999, 195 patients with ages ranging from 12 to 70 years and presenting “de novo” AML,excluding the M3 subtype. In the eighties, 102 patientsare on record: 47 males and 55 females. Among these,84 received induction chemotherapy with Cytarabinplus Daunorrubicin (7+3), resulting in a 51% (43/84) remission rate. The average disease-free andoverall survival was 35% at ten months and 13% at 12 months. Forty-one new diagnoses were performedfrom January 1990 and December 1993, all patients were submitted to the “7+3” chemotherapy protocoland a 66% remission rate was obtained. The disease free and overall survival rates were statistically (p <0.001) higher with 41% of the patients at 7.6 months and 17.5% at 22 months. In 1994 a new protocol wasintroduced under the guidance of the Brazilian Cooperative Group on Leukemia Studies (GCBEL). Itrecommended induction with Cytarabin +Idarrubicin followed by consolidation and two intensification cycles with high doses of Cytarabin................................................... We observed a progressive improvement in remission and survival rates in the last decades, however the prolonged eventfree survival stayed below 30%. Our results are similar to the national centers that treat AML and inaccordance with the literature data.

Níveis de hematócrito e hemoglobina e estado nutricional em crianças de 7 a 12 anos em um bairro de baixa renda de Porto Alegre, RS / Levels of hematocrit and hemoglobin and nutritional status in children from 7 to 12 years old in a low income population district of Porto Alegre

O presente trabalho avaliamos 128 crianças de 7 a 12 anos quanto a níveis de hematócrito e hemoglobina, prevalência de valores baixos de hematócrito e hemoglobina, estado nutricional conforme critérios de Seoane Lathan. A populaçäo era de baixa renda com um salário médio mensal entre 1 e 2 salários mínimos, que correspondem a US$50 e US4100 respectivamente. Os níveis médios de hematócritos e hemoglobina encontrados foram de 36.37 (+/- 2.58) e 10.87 (+/- 1.28) respectivamente. A prevalência de hematócrito menor que 34% de hemoglobina menor que 11 g/dl foi de 10,9% e 53.9% respectivamente. Da populaçäo, 89,1% apresentavam algum tipo de desnutriçäo conforme critérios de Seoane Lathan, sendo a mais freqüente Desnutriçäo Crônica Atual