Philadelphia-negative myeloproliferative neoplasms as disorders marked by cytokine modulation

Background: Cytokines are key immune mediators in physiological and disease processes, whose increased levels have been associated with the physiopathology of hematopoietic malignancies, such as myeloproliferative neoplasms. Methods: This study examined the plasma cytokine profiles of patients with essential thrombocythemia, primary myelofibrosis, polycythemia vera and of healthy subjects, and analyzed correlations with JAK2 V617F status and clinical-hematological parameters. Results: The proinflammatory cytokine levels were increased in myeloproliferative neoplasm patients, and the presence of the JAK2 V617F mutation was associated with high IP-10 levels in primary myelofibrosis patients. Conclusions: Essential thrombocythemia, primary myelofibrosis, and polycythemia vera patients exhibited different patterns of cytokine production, as revealed by cytokine network correlations. Together, these findings suggest that augmented cytokine levels are associated with the physiopathology of myeloproliferative neoplasms.

Deregulation of apoptosis-related genes is associated with PRV1 overexpression and JAK2 V617F allele burden in Essential Thrombocythemia and Myelofibrosis

<p>Abstract</p> <p>Background</p> <p>Essential Thrombocythemia (ET) and Primary Myelofibrosis (PMF) are Chronic Myeloproliferative Neoplasms (MPN) characterized by clonal myeloproliferation/myeloaccumulation without cell maturation impairment. The JAK2 V617F mutation and <it>PRV1 </it>gene overexpression may contribute to MPN physiopathology. We hypothesized that deregulation of the apoptotic machinery may also play a role in the pathogenesis of ET and PMF. In this study we evaluated the apoptosis-related gene and protein expression of BCL2 family members in bone marrow CD34⁺hematopoietic stem cells (HSC) and peripheral blood leukocytes from ET and PMF patients. We also tested whether the gene expression results were correlated with JAK2 V617F allele burden percentage, <it>PRV1 </it>overexpression, and clinical and laboratory parameters.</p> <p>Results</p> <p>By real time PCR assay, we observed that <it>A1, MCL1, BIK and BID</it>, as well as <it>A1, BCLW </it>and <it>BAK </it>gene expression were increased in ET and PMF CD34⁺cells respectively, while pro-apoptotic <it>BAX </it>and anti-apoptotic <it>BCL2 </it>mRNA levels were found to be lower in ET and PMF CD34⁺cells respectively, in relation to controls. In patients' leukocytes, we detected an upregulation of anti-apoptotic genes <it>A1, BCL2, BCL-X_L</it>and <it>BCLW</it>. In contrast, pro-apoptotic <it>BID </it>and <it>BIM_EL</it>expression were downregulated in ET leukocytes. Increased BCL-X_Lprotein expression in PMF leukocytes and decreased BID protein expression in ET leukocytes were observed by Western Blot. In ET leukocytes, we found a correlation between JAK2 V617F allele burden and <it>BAX, BIK and BAD </it>gene expression and between <it>A1, BAX </it>and <it>BIK </it>and <it>PRV1 </it>gene expression. A negative correlation between <it>PRV1 </it>gene expression and platelet count was observed, as well as a positive correlation between <it>PRV1 </it>gene expression and splenomegaly.</p> <p>Conclusions</p> <p>Our results suggest the participation of intrinsic apoptosis pathway in the MPN physiopathology. In addition, <it>PRV1 </it>and JAK2 V617F allele burden were linked to deregulation of the apoptotic machinery.</p

Deregulated expression of apoptomirs and apoptosis-related genes and proteins in Primary Myelofibrosis and Essential Thrombocythemia

As Neoplasias Mieloproliferativas Crônicas (NMPC) Trombocitemia Essencial (TE), Mielofibrose (MF) - são desordens hematopoéticas resultantes da expansão clonal da célula tronco hematopoética alterada. Essas doenças caracterizam-se pela independência dos progenitores hematopoéticos aos estímulos dos fatores de crescimento e citocinas e pela proliferação exacerbada das células da linhagem mielóide com maturação preservada. Os mecanismos moleculares e celulares envolvidos na patogênese e progressão da TE e MF não foram ainda esclarecidos, mas o mieloacúmulo presente nessas doenças parece estar associado à alteração de proliferação e apoptose celular. Nesse contexto, os objetivos deste trabalho foram avaliar em células CD34+ da medula óssea e leucócitos dos pacientes com TE e MF: (1) a expressão de apoptomirs e de genes pró- e anti-apoptóticos pertencentes à via intrínseca e extrínseca da apoptose pela metodologia de RT-PCR em tempo real; (2) expressão de proteínas pró- e anti-apoptóticas por Western-blot; (3) o perfil de sensibilidade das células mononucleares à apoptose induzida por diferentes agentes apoptogênicos pela técnica de citometria de fluxo e (4) correlacionar os resultados obtidos com dados clínico-laboratoriais dos pacientes e com a expressão do gene PRV-1. Em TE, nas células CD34+, foi detectado aumento da expressão de a1, mcl-1, bid, bok e noxa e diminuição de bax em relação aos controles. Nos leucócitos, foi detectada a elevação da expressão de a1, bcl-2, bcl-w e bcl-xL, bad e bok e diminuição de bid e bimEL. Em comparação com o grupo controle, os pacientes com MF apresentaram maior expressão dos genes a1, bcl-w bak, bok e noxa e menor de bcl-2 nas células CD34+. Nos leucócitos dos pacientes com MF foi verificado aumento da expressão dos genes bcl-2, bcl-w e bcl-xL, bad e bok. O c-iap-1 apresentou maior expressão nas células CD34+ dos pacientes com MF e TE, enquanto que o c-iap-2 estava elevado nos leucócitos e células CD34+. Foi ainda detectada a expressão diferencial em TE e MF dos genes pertencentes a via de receptor de morte, como a maior expressão dos genes fas, fas-L, faim e dr4 nas células CD34+ dos pacientes e menor expressão do fas-L e trail nos leucócitos dos pacientes com TE e MF. Os resultados indicaram associação da expressão do gene PRV-1 com a mutação JAK2V617F e com a expressão dos genes a1, bcl-w, bik, bax, c-iap-2 e trail. Com relação à expressão proteica, BCL-XL estava aumentada e TRAIL diminuída em leucócitos de pacientes com MF enquanto que a molécula BID estava diminuída em leucócitos de pacientes com TE. Os miRNA26a, let 7d e miR15a estavam mais expressos nos leucócitos de pacientes com TE e MF. Em pacientes com TE o miR130b estava elevado e o mIR16 diminuído, enquanto que nos pacientes com MF o miR198 estava aumentado. Os linfócitos dos pacientes com TE e MF apresentaram maior resistência à apoptose estimulada por: actinomicina, etoposídeo, teniposídeo, citarabina e estaurosporina do que os linfócitos dos controles. Em conclusão, os dados obtidos sugerem a ligação da alteração do processo de apoptose celular, com a expressão do gene PRV-1 e status da mutação JAK2V617F. Esses resultados contribuem para o melhor entendimento da fisiopatologia das NMPC visto que associam a fisiopatologia da TE e MF com a resistência das células alteradas à apoptose. Essas informações serão úteis no futuro, possibilitando o desenho de novos alvos terapêuticos e a descrição de novos marcadores de prognóstico.Chronic Myeloproliferative Neoplasms (cMPN) - Essential Thrombocythemia (ET), Primary Myelofibrosis (PMF) and Polycythemia Vera (PV) - are clonal hematopoietic stem cell malignancies characterized by an accumulation of mature myeloid cells in bone marrow and peripheral blood. It seems that apoptotic machinery deregulation contribute to ET and PMF pathogenesis. Despite the advances in the molecular knowledge, the physiopathology of these diseases remains unknown. This study investigated cellular and molecular mechanisms involved in apoptosis process regulation in bone marrow haematopoietic progenitor CD34+ cells and leukocytes from ET and PMF patients. The specifics aims were: (1) to evaluate death receptors family members and Bcl-2 related genes expression as well apoptosis-related microRNAs by Real Time PCR; (2) apoptosis-related protein expression by Western Blot; (3) access mononuclear cells apoptosis resistance by flow cytometry and (4) to correlate the results with JAK2V617F mutation, PRV-1 gene expression and as well clinical data. In ET CD34+ cells, we found overexpression of a1, mcl-1, bid, bok e noxa and a decrease of bax compared to controls, while in leukocytes a1, bcl-2, bcl-w e bcl-xL, bad e bok expression was increased and bid and bimEL expression was lower than in controls. In PMF, a1, bcl-w bak, bok e noxa were overexpressed and bcl-2 downregulated in CD34+ cells. In PMF leukocytes bcl-2, bcl-w e bcl-xL, bad e bok mRNA levels were increased. c-iap-1 was increased in ET and PMF CD34+ cells and c-iap-2 expression elevated in ET and PMF CD34+ cells and leukocytes. Death receptor related genes showed overexpression of fas, fas-L, faim and dr4 in patients CD34+ cells and downregulation of fas-L and trail in ET and PMF leukocytes. We found differential expression of several genes between patients JAK2V617F positive and negative, as well we found correlation between gene expression and JAK2V617F allele burden, white blood cells and platelets count and splenomegaly. PRV-1 gene was overexpressed in ET and PMF leukocytes and showed correlation with JAK2V617F mutation and a1, bcl-w, bik, bax, c-iap-2 and trail gene expression. Regarding protein expression, BCL-XL was increased and TRAIL decreased in PMF leukocytes and BID was decreased in ET leukocytes. miRNA26a, let 7d e miR15a was overexpressed in ET and PMF leukocytes, while miR130b was increased only in ET and miR198 only in PMF. miR16 was downregulated in ET leukocytes comparing to controls. We also detected a resistance to apoptosis-inducers in ET and PMF lymphocytes and we observed correlation between apoptosis percentage and the expression of many studied genes. In conclusion, the results indicate the participations of Bcl-2 family genes and Death Receptor pathway genes, as well PRV-1 and JAK2V617F mutation in these disorders, which contribute to elucidate cMPN physiopathology and might lead to the discovery of new cMPN therapies and molecular markers

Frequência de consumo de alimentos fonte de ferro entre crianças de 6 a 59 meses atendidas pela Estratégia de Saúde da Família

Introduction: The introduction of adequate complementary feeding and the formation of healthy eating habits are major challenges in Brazil, where many important foods for children's development are not offered to them due to regional beliefs, taboos and cultures; as well as access and availability. Some of these foods act directly in the prevention of iron deficiency anemia, such as meat and offal, green leaves in general; beans and egg yolk. Objective: This study aimed to evaluate the frequency of consumption of iron source foods among children enrolled in the Family Health Strategy. Material and Methods: A cross-sectional study was conducted with 256 children aged 6-59 months enrolled in 10 units of the Family Health Strategies in a city in the state of Minas Gerais. A structured questionnaire was applied and anthropometric evaluation was performed. Mann-Whitney test and Pearson's chi-square test were used to compare the results according to the child's age, adopting p &lt;0.05. Results: The iron source foods most consumed were beans (79.7%), bakery products (78.9%) and flour for children (29.9%), while the other nine foods measured had a rare frequency of consumption. Older children had a higher frequency of consumption for leafy vegetables, red meats, poultry and eggs, while the younger ones (&lt;24 months) higher frequency of consumption for milk formulas, indicating the belated introduction of iron source foods. Conclusion: It has been identified low frequency of consumption of food source of iron for children, providing the risk for nutritional deficiency of iron, reinforcing the importance of family guidance regarding the diversification of the child in order to contribute to the promotion of health and prevention of deficiencies such as iron.Introdução: A introdução da alimentação complementar de forma adequada e a formação de hábitos alimentares saudáveis são grandes desafios no Brasil, onde muitos alimentos importantes para o desenvolvimento das crianças não lhe são ofertados devido a crenças, tabus e culturas regionais, bem como acesso e disponibilidade. Alguns desses alimentos atuam diretamente na prevenção da anemia ferropriva, como carnes e vísceras, folhas verdes em geral, feijões e gema de ovo. Objetivo: Avaliar a frequência de consumo de alimentos fonte de ferro entre crianças de 6 a 59 meses cadastradas na Estratégia de Saúde da Família. Material e Métodos: Trata-se de estudo transversal realizado com 256 crianças de 6 a 59 meses cadastradas na Estratégia de Saúde da Família de um município no interior do estado de Minas Gerais. Aplicou-se questionário estruturado sobre condições de vida e saúde e frequência alimentar e avaliaram-se dados antropométricos. Utilizaram-se testes de Mann-Whitney e qui-quadrado de Pearson na comparação dos resultados segundo faixa etária da criança, adotando-se p&lt;0,05. Resultados: Os alimentos fonte de ferro mais consumidos foram feijões (79,7%), produtos de padaria (78,9%) e farinhas infantis (29,9%), sendo o consumo dos outros nove alimentos avaliados classificados como raro. Crianças mais velhas apresentaram maior frequência de consumo para vegetais folhosos, carnes vermelhas, aves e ovos, sendo que nas crianças com menos de 24 meses as fórmulas lácteas apresentaram maior frequência, demonstrando introdução tardia dos alimentos fonte de ferro. Conclusão: Foi identificada baixa frequência de consumo dos alimentos fonte de ferro pelas crianças, oportunizando a carência nutricional de ferro, reforçando a importância da orientação da família quanto à diversificação da alimentação da criança de forma a contribuir para a promoção de saúde e prevenção de deficiências como a de ferro. &nbsp

Interaction between FAK/αB-crystalline is important for viability of the glioblastoma cells

Abstract Glioblastoma multiforme is a tumor of the central nervous system. Focal Adhesion Kinase (FAK) and αB-crystalline are two proteins involved in glioblastoma development. In this study, we investigated whether the FAK/αB-crystalline interaction is important for glioblastoma cells, we aimed to investigate the interaction of these two proteins in the glioblastoma multiforme cell line U87-MG. Two peptides named FP01 peptide (derived from αB-crystalline) and FP02 peptide (derived from FAK) were synthesized for this study. Treatment of U87-MG with the peptides FP01 and FP02 in the concentration at 50 µM reduced the viability cellular to around 41% and 51%, respectively. Morphological alterations in the cells treated with the peptides when compared to the control were observed. This study suggests that the interaction between FAK and αB-crystalline is important for the viability of glioblastoma cells

Risk factors associated with neonatal thrombosis: scope review protocol

Scoping review of the indexed literature on neonatal thrombosis and the associated risk factors

Bullying in elderly people

OBJECTIVE: To report the case of a patient who suffered bullying in a long-term care facilities. METHOD: The needed data was obtained by a medical chart review, interview with the patient, and literature review. RESULTS: The reported case and the data reviewed bring to focus violence suffered by elderly people, especially, about the bullying that is a form of violence too little researched in elderly people. There is a need of further researches about the topic, focusing on prevention and early detection.</p

Apoptosis- and cell cycle-related genes methylation profile in myeloproliferative neoplasms

Department of Clinical, Toxicological and Bromatological Analyses, School of Pharmaceutical Sciences of Ribeirão Preto , University of São Paulo , Ribeirão Preto , SP , BrazilDepartment of Pharmacy, School of Pharmacy , Federal University of Juiz de Fora, Governador Valadares Campus , Governador Valadares , MG , BrazilEuryclides de Jesus Zerbini Transplant Hospital , São Paulo , SP , BrazilDepartment of Internal Medicine , Ribeirão Preto Medical School, University of São Paulo , Ribeirão Preto , SP , BrazilDivision of Hematology , Federal University of São Paulo , São Paulo , SP , BrazilResearcher of the National Council for Scientific and Technological Development (CNPq) , Brasília , DF , BrazilFleury Institute , São Paulo , SP , BrazilDivision of Hematology , Federal University of São Paulo , São Paulo , SP , BrazilWeb of Scienc