The possibility of early diagnosis of colorectal cancer through expression of Lncrna UCA1, Lncrna LINC00970, and Wnt genes; an in-vitro study

Colorectal cancer is the 4thcause of cancer death; with considering the growth process of this cancer and the necessity of early diagnosis, the purpose of the research is to state the LncRNA 00970, LncRNA UCAI,and the Wntgene before and after the treatment by 5-Azacytidine epigenetic medicine, to reach the biomarker in the very first steps of colorectal cancer. In this experiment, the human colon cancer cell line (HT29) treated with different concentrations of 5-aza-2'-deoxycytidine (5-aza-dC) was utilized to induce DNA demethylation; Quantitative PCR (qPCR) was used to measure LncRNA UCA1and LncRNA LINC00970 and Wntexpression. There was a significant relationship between the expression of LncRNA 00970, LncRNA UCAI,and the Wntgene and its effects on colorectal (p < 0.05). The Wntgene was treated by 1 and 10 of 5-Azacytidine epigenetic medicine, which then experienced decreases. In LncRNA UCAI and LncRNA00970 in dose 1 micromolar of 5-Azacytidine had decrement and increment of expressionrespectively that explains their efficiency but in treatment by dose 10 mM of this medicine, no significant LncRNA expression difference was detected, 5-azacitidine has a direct impact on its target genes and LncRNAs.Therefore, it can be used in the early diagnosis of colorectal cancer.

Pioderma gangrenoso ampollar como forma de presentación de leucemia mieloide agudo: informe de un caso / Acute myeloid leukemia presented in the form of bullous pyoderma gangrenosum: a case report

El pioderma gangrenoso ampollar es una variedad infrecuente de pioderma gangrenoso, que se asocia en el 50-70% de los casos con trastornos oncohematológicos. Se comunica el caso de una paciente de 59 años, que consultó por fiebre y ampollas purpúricas de rápida progresión, con compromiso cutáneo mucoso. Con sospecha de una enfermedad neutrofílica, ampollar, o infección por gérmenes oportunistas, se realizó biopsia de piel para estudio histopatológico, inmunofluorescencia directa y cultivo. Los cultivos y la inmunofluorescencia directa fueron negativos, y la anatomía patológica reveló un denso infiltrado inflamatorio con predominio neutrofílico en dermis. Ante el diagnóstico de pioderma gangrenoso ampollar, se realizó una punción-aspiración de médula ósea cuyo resultado fue compatible con leucemia mieloide aguda. Se instauró tratamiento con corticosteroides sistémicos, a pesar de lo cual la paciente evolucionó desfavorablemente y falleció a los 15 días de su ingreso hospitalario. Este caso ilustra la asociación de esta enfermedad cutánea con trastornos oncohematológicos y el mal pronóstico que esto implica a corto plazo. (AU)

Bullous pyoderma gangrenosum is an infrequent type of pyoderma gangrenosum, associated with onco hematological diseases in 50-70% of cases. We present the case of a 59-year-old patient with fever and mucocutaneous hemorrhagic bullous of rapid progression. A biopsy for histopathology, direct immunofluorescence (DIF) and skin culture was made, considering the possibility of neutrophilic dermatoses, bullous dermatosis or an opportunistic infection. The results of both the culture and the DIF were negative. The histopathological examination of the specimen revealed a dense dermal polymorphic infiltrate composed primarily of neutrophils. Considering bullous pyoderma gangrenosum as a potential diagnosis, a bone-marrow biopsy was performed. This study revealed an acute myeloid leukemia. Although systemic corticosteroid therapy was begun, the patient presented an unfavorable evolution that led to her death 15 days after her admission at the hospital. This case shows the association between bullous pyoderma gangrenosum and onco hematological diseases. In addition, it highlights the poor prognosis related to these diseases in the short term. (AU)

5-Azacytidine restores interleukin 6-increased production in mesenchymal stromal cells from myelodysplastic patients

Introduction: Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematological diseases. In addition to defects in hematologic progenitor and stem cells, dysfunctions in the bone marrow microenvironment (BMM) participate in the MDS pathogenesis. Furthermore, the immune response is deregulated by the pro-inflammatory response prevailing in low-risk MDS, while immunosuppression predominates in high-risk MDS. Mesenchymal stromal cells (MSC), part of the BMM, are characterized by plastic adherent growth and multipotentiality. They exhibit immunomodulatory properties and sustain hematopoiesis. There is conflicting evidence regarding their status in MDS. The aim of this study was to characterize MDS-MSC and evaluate the effect of 5-Azacytidine. Methods: The MSC from MDS patients and controls were cultured and characterized according to the International Society of Cell Therapy recommendations. Immunomodulatory properties were assessed by studying the MSD cytokine production, using the cytometric bead array. We evaluated the effect of 5-Azacytidine on the MSC cytokine production. Results: We included 35 MDS patients and 22 controls. The MSC from patients and controls were cultured and characterized. The MSC from patients showed morphological differences, but there were no differences in immunophenotype or multipotentiality. The interleukin 6 (IL-6) was the main MSC secreted cytokine. The MDS-MSC produced higher levels of IL-6, IL-17, interferon gamma, or interferon γ (INF-γ), and tumor necrosis factor alpha (TNF-α). The in vitro 5-Azacytidine treatment induced a significant decrease in the IL-6 production by MDS-MSC. Conclusions: The MDS-MSC show an increased production of pro-inflammatory cytokines. The in vitro treatment with 5-Azacytidine lead to a significant reduction in the IL-6 production by the MDS-MSC, restoring the IL-6 levels to those found in controls. The MSC produced inflammatory cytokines involved in the MDS pathogenesis, representing a potential future therapeutic target. Moreover, 5-Azacytidine may have a stromal effect, modulating the immune response in MDS.

Targeted therapy with a selective BCL-2 inhibitor in older patients with acute myeloid leukemia

ABSTRACT Background: Older patients with acute myeloid leukemia are particularly difficult to treat, as they have a high risk of comorbidities, poor performance status and less tolerability to chemotherapy, as well as a more aggressive disease biology, responsible for the resistance to treatment. There is a need to explore novel therapeutic agents that are more effective and tolerable. Venetoclax, a BCL-2 inhibitor is a promising agent, as BCL-2 overexpression is present in 84% of acute myeloid leukemia patients at diagnosis and 95% of patients at relapse and has been associated with leukemia cell survival, chemotherapy resistance and poor prognosis. Objective: To review the available data about venetoclax in acute myeloid leukemia and how it can influence the treatment in older patients. Methods: Using the Pubmed database, we selected 29 articles published within the last 15 years, considering preclinical and clinical trials and review studies that combined venetoclax with acute myeloid leukemia. Results: Venetoclax has demonstrated promising results in preclinical and clinical trials, especially in patients with poor prognosis and the IDH mutation, with an excellent side-effect profile. However, resistance seems to develop rapidly with venetoclax monotherapy, because of antiapoptotic escape mechanisms. Conclusions: While the results with the use of venetoclax seem encouraging, it is not likely that targeting a single pathway will result in long-term disease control. The solution includes the use of combined therapy to block resistance mechanisms and enhance apoptosis, by reducing MCL-1, increasing BIM or inhibiting the complex IV in the mitochondria.

Trasplante de médula ósea alogénico en un paciente con fusariosis diseminada y leucemia mieloide aguda / Allogeneic hematopoietic cell transplantation in a patient with disseminated fusariosis and acute myeloid leukemia

La infección diseminada por Fusarium se ha convertido en un problema creciente en las personas con neoplasias hematológicas malignas, principalmente en pacientes con leucemias agudas; se describen cada vez más casos en aquellos sometidos a un trasplante de médula ósea. No existe un tratamiento óptimo establecido para la fusariosis diseminada. La mortalidad global comunicada de esta infección oscila entre el 50 y el 80%. Se presenta a continuación el caso de un paciente de sexo masculino de 29 años, con diagnóstico de leucemia mieloide aguda, que presenta como complicación una fusariosis diseminada, y logra sobrellevar un trasplante alogénico de médula ósea en el Hospital Italiano de San Justo (Argentina) de forma exitosa. (AU)

Disseminated fusariosis has become an increasing problem in people with hematopoietic neoplasms, mainly in patients affected by acute leukemias, and even more in those who undergo hematopoietic cell transplantation. There is not an optimal treatment for disseminated fusariosis. The global mortality described in the literature is between 50% and 80%. We introduce a case of a 29 year old patient with diagnosis of acute myeloid leukemia complicated with disseminated fusariosis, who copes with an allogeneic hematopoietic cell transplantation with a successful outcome in the "Hospital Italiano de San Justo" (Argentina). (AU)

Successful treatment of post-transplant relapsed acute myeloid leukemia with FLT3 internal tandem duplication using the combination of induction chemotherapy, donor lymphocyte infusion, sorafenib and azacitidine. Report of three cases / Tratamento bem-sucedido de leucemia mieloide aguda recorrente após transplante com duplicação interna em tandem FLT3 usando combinação de indução por quimioterapia, infusão de linfócitos de doador, soferanib e azacitidina. Relato de três casos

ABSTRACT Acute myeloid leukemia is a hematopoietic stem cell neoplastic disease associated with high morbidity and mortality. The presence of FLT3 internal tandem duplication mutations leads to high rates of relapse and decreased overall survival. Patients with FLT3 internal tandem duplication are normally treated with hematopoietic stem cell transplantation in first complete remission. Nevertheless, the incidence of post-transplant relapse is considerable in this group of patients, and the management of this clinical condition is challenging. The report describes the outcomes of patients with FLT3 internal tandem duplication positive acute myeloid leukemia who relapsed after allogeneic hematopoietic stem cell transplantation and were treated with the combination of re-induction chemotherapy, donor lymphocyte infusion, sorafenib and azacitidine. Three cases are described and all patients achieved prolonged complete remission with the combined therapy. The combination of induction chemotherapy followed by donor lymphocyte infusion, and the maintenance with azacitidine and sorafenib can be effective approaches in the treatment of post-hematopoietic stem cell transplant and relapsed FLT3 internal tandem duplication positive acute myeloid leukemia patients. This strategy should be further explored in the context of clinical trials.

RESUMO A leucemia mieloide aguda é uma doença neoplásica de células-tronco hematopoiéticas com alta morbimortalidade. A presença de mutações de duplicação em tandem de FLT3 leva a altas taxas de recorrência e a menor sobrevida global. Os pacientes com duplicação em tandem de FLT3 são normalmente tratados com transplante de células-tronco hematopoiéticas na primeira remissão completa. No entanto, a incidência de recidiva pós-transplante é considerável neste grupo de pacientes, e a conduta, nestes casos, é um desafio. O relato descreve os resultados do tratamento de pacientes com leucemia mieloide aguda positiva e duplicação em tandem de FLT3 que recidivaram depois do transplante alogênico de células-tronco hematopoiéticas e que foram tratados com combinação de quimioterapia de reindução, infusão de linfócitos de doador, sorafenib e azacitidina. São descritos três casos, e todos os pacientes apresentaram remissão completa prolongada com a terapia combinada. A combinação de quimioterapia de indução, seguida de infusão de linfócitos do doador, e a manutenção com azacitidina e sorafenib podem ser abordagens eficazes no tratamento da recorrência pós-transplante em pacientes com leucemia mieloide aguda e duplicação em tandem de FLT3. Essa estratégia deve ser mais explorada no contexto de ensaios clínicos.

Eficacia y seguridad del uso de azacitidina en el tratamiento de síndrome mielodisplásico de bajo riesgo o intermedio-i con alto requerimiento transfusional con falla al tratamiento con primera línea / Efficacy and safety of the use of azacitidine in the treatment of low-risk or intermediate-i myelodysplastic syndrome with high transfusion requirement with failure of first-line treatment

INTRODUCCIÓN: Antecedentes: El presente dictamen expone la evaluación de tecnología de la eficacia y seguridad de azacitidna en el tratamiento de síndrome mielodisplásico de bajo riesgo o internedio-i con alto requerimientp transfusional con falla al tratamientop con primera linea. Aspectos Generales: En el sistema de clasificación de cánceres hematológicos de la Organización Mundial de la Salçud, los síndromes mielodisplásicos representam una de las cinco categorais principales de neoplasias mieloides. Es un trastorno hematológico caracteriazado por hematopoyesis ineficaz, una o más citopenias, y con frecuencia médula ósea hipercelular, además de una predisposición inherente a transformarse en leucemia mieloide aguda. Tecnologia Sanitaria de Interés: La Azacitidina es un cucleósido de pirimidina, análogo de citidina. Inhibe la ADN metiltransferada, que es la enzima responsable de metilar el ADN recientemente sintetizado, la hipometilación resultante en el ADN conlleva cambios en la transcripción y expressión de algunos genes. METODOLOGIA: Estrategia de Búsqueda: Se realizó una busqueda de literatura publicada sobre Azacitidina en el tratamiento de Síndrome Mielodisplásico de riesgo bajo o intermedio-I en la bases de datos: Medline y Tripdatabase. Adicionalmente, se realizaron búsquedas en los portales web de entidades que realizan revisiones sistemáticas, evaluaciones de tecnologías sanitarias y guías de práctica clínica: The Cochrane Library, National Institute for Health and Care Excelence (NICE) del Reino Unido, national Guideline of Clearinghouse, y National Comprehensive Cancer Network (NCCN) de los Estados Unidos, y The Scottish Intgercollegiate Network (SIGN) de Escocia. RESULTADOS: Sinopsis de la evidencia: Se realizó la búsqueda y la revisión de la evidencia acorde a lo estipulado en la pregunta PICO formulada. Así se consideraron los estudios que tuvieran como intervención azacitidina en el tratamiento de síndrome mielodisplásico de bajo riesgo o intermedio-i con alto requerimiento transfusional con falla al tratamiento con primera línea. CONCLUSIONES: La presente evaluación de tecnología tuvo por objeitvo la evaluación de la eficacia y seguridad del uso de azacitidina en el tratamiento de síndrome mielodisplásico de bajo riesgo o intermedio-i con alto requerimiento transfusional con falla al tratamiento con primera línea. No se ha encontrado en la presente evaluación de tecnología sanitaria evidencia consistente que establezca cual es el beneficio neto atribuible al uso de azacitidina por sobre la mejor terapia de soporte en pacientes con síndrome mielodisplásico, con IPSS de riesgo bajo o intermedio-1, alto requerimiento transfusional, y resistencia o falla a los agentes estimulantes de hematopoyesis. Considerando que la principales guías recomiendan en estos pacientes a la terapia de soporte, las transfusiones, los agentes estimulantes de eritropoyesis, inmunosupresores (en candidatos apropiados), y lenalodomida en pacientes con deleción 5a; se debe mantener estas recomendaciones de tratamiento hasta que futuros ensayos clínicos que comparen azacitidina versus terapia de soporte en esta población, informen concluyentemente sobre la eficacia y seguridad de su uso. El Instituto de Evaluación de Tecnologías e Investigación - IETSI no aprueba el uso de azacitidina como una alternativa de tratamiento para pacientes con síndrome mielodisplásico, con IPSS de riesgo bajo o intermedio-1, alto requerimiento transfusional, y resistencia o falla a los agentes estimulantes de hematopoyesis.

Caracterización clínica de los pacientes con síndrome mielodisplásico / Clinical characterization of patients with myelodysplastic syndrome

Objetivo: Describir las características clínicas, sociodemográficas y calidad de vida de los pacientes con síndrome mielodisplasico (MDS). Diseño: Estudio descriptivo de corte transversal. Marco de referencia: Hospital San José de Bogotá (institución de cuarto nivel). Población: Treinta y nueve pacientes con diagnóstico de MDS, mayores de 18 años, obtenidos en el departamento de estadística. Mediciones principales: Frecuencia de variables clínicas, sociodemográficas y calidad de vida medida por la escala FACT-G. Resultados: La edad promedio fue de 74 años, con un predominio en hombres. El diagnóstico más frecuente fue MDS tipo displasia mutilinaje, y la citopenia más prevalente fue anemia. La mayoría de los pacientes fue clasificado como muy bajo riesgo o bajo riesgo por la escala IPSS-R y WPSS, y bajo riesgo y riesgo intermedio -1 por la escala IPSS y MDACC. La progresión a leucemia mieloide aguda se presentó en 7.69%, mientras que la mortalidad en 15.38%. Existió una mayor frecuencia de muerte y transformación a leucemia en los pacientes clasificados como de alto riesgo. La calidad de vida fue inferior en los pacientes con anemia y fue superior en el MDS-(del5q). Conclusiones: Las características de la población estudiada son similares a las reportadas en la literatura (predominio en hombres, subtipo de MDS más frecuente, citopenia predominante y distribución de riesgo). Existieron diferencias con respecto a la edad, frecuencia de comorbilidades y su relación con mortalidad. La calidad de vida se correlacionó con el subtipo de MDS y citopenias. (Acta Med Colomb 2016; 41: 36-41).

Objective: To describe the clinical and socio-demographic characteristics and quality of life of patients with myelodysplastic syndrome (MDS). Design: a descriptive cross-sectional study. Framework: Hospital San José de Bogota (fourth level institution). Population: Thirty-nine patients diagnosed with MDS, older than 18 years, obtained in the statistics department. Main measurements: frequency of clinical and sociodemographic variables and quality of life measured by FACT-G scale. Results: The mean age was 74 years, with male predominance. The most frequent diagnosis was type mutilineage MDS dysplasia, and the most prevalent cytopenia was anemia. Most patients were classified as very low risk or low risk by IPSS-R and WPSS scale and low-risk and intermediate risk -1by IPSS and MDACC scale. Progression to acute myeloid leukemia occurred in 7.69%, while mortality occurred in 15.38%. There was a higher incidence of death and transformation to leukemia in patients classified as high risk. The quality of life was lower in patients with anemia and was higher in the MDS-(del 5q). Conclusions: The characteristics of the study population are similar to those reported in the literature (predominantly in men, subtype of MDS more frequent, predominant cytopenia and risk distribution). There were differences with respect to age, frequency of comorbidities and their relationship to mortality. Quality of life was correlated with the subtype of MDS and cytopenias. (Acta Med Colomb 2016; 41: 36-41).

Déficit adquirido de factor X en leucemia mielomonocítica crónica: reporte de un caso / Isolated factor X deficiency in chronic myelomonocytic leukemia: Report of one case

Bleeding disorders are commonly associated with hemato-oncologic diseases. We report a 68 years old male with a chronic myelomonocytic leukemia derived from a long lasting mielodysplastic syndrome that did not respond to treatment with Azacitidine. The patient was hospitalized due to tonic clonic seizures. A CAT scan showed a hematoma in the frontal lobe. A new assessment of hemostasis revealed an isolated deficiency of Factor X. We speculate that this deficit could be secondary to consumption due to the chronic Myelomonocytic Leukemia.

Azacitidine and lenalidomide as an alternative treatment for refractory acute myeloid leukemia: a case report / Azacitidina e lenalidomida como alternativa de tratamento para leucemia mieloide aguda refratária: um relato de caso

CONTEXT: Refractory acute myeloid leukemia (AML) is a difficult disease to control with second or third-line chemotherapy regimens. In this report, we describe using azacitidine in combination with lenalidomide as salvage therapy. CASE REPORT: 52-year-old female was diagnosed with refractory AML and high-risk cytogenetics: complex monosomal karyotype consisting of t (3, 3) in association with monosomy 7 and del 5q. Morphological remission associated with maintenance of the cytogenetic abnormality of chromosome 3 and disappearance of the abnormalities relating to chromosomes 5 and 7 was achieved after three cycles of combination therapy with azacitidine and lenalidomide. CONCLUSION: Azacitidine plus lenalidomide can be a therapeutic option for patients with refractory AML, as illustrated in this case. .

CONTEXTO: A leucemia mieloide aguda (LMA) refratária é considerada doença de difícil controle com regime quimioterápico de segunda ou terceira linha. Neste relato, é descrito o uso de azacitidina em combinação com lenalidomida como esquema de resgate. RELATO DE CASO: Paciente de 52 anos, do sexo feminino, com o diagnóstico de LMA refratária de alto risco citogenético, apresentava cariótipo complexo e monossômico, com t (3, 3), associado à monosomia do 7 e del 5q. Destaca-se que, após três ciclos da terapia combinada com azacitidina e lenalidomida, houve remissão morfológica, com manutenção da anormalidade citogenética relacionada ao cromossomo 3 e desaparecimento da anormalidade relacionada aos cromossomos 5 e 7. CONCLUSÃO: Azacitidina e lenalidomida podem ser opção terapêutica para pacientes com LMA refratária, como demonstrado neste caso. .

Stability of XIST repression in relation to genomic imprinting following global genome demethylation in a human cell line

DNA methylation is essential in X chromosome inactivation and genomic imprinting, maintaining repression of XIST in the active X chromosome and monoallelic repression of imprinted genes. Disruption of the DNA methyltransferase genes DNMT1 and DNMT3B in the HCT116 cell line (DKO cells) leads to global DNA hypomethylation and biallelic expression of the imprinted gene IGF2 but does not lead to reactivation of XIST expression, suggesting that XIST repression is due to a more stable epigenetic mark than imprinting. To test this hypothesis, we induced acute hypomethylation in HCT116 cells by 5-aza-2′-deoxycytidine (5-aza-CdR) treatment (HCT116-5-aza-CdR) and compared that to DKO cells, evaluating DNA methylation by microarray and monitoring the expression of XIST and imprinted genes IGF2, H19, and PEG10. Whereas imprinted genes showed biallelic expression in HCT116-5-aza-CdR and DKO cells, the XIST locus was hypomethylated and weakly expressed only under acute hypomethylation conditions, indicating the importance of XIST repression in the active X to cell survival. Given that DNMT3A is the only active DNMT in DKO cells, it may be responsible for ensuring the repression of XIST in those cells. Taken together, our data suggest that XIST repression is more tightly controlled than genomic imprinting and, at least in part, is due to DNMT3A.

Uso tópico de 5-azacitidina melhora a cicatrização de feridas cutâneas de roedores por meio do sistema ativina/folistatina / Topical 5-azacytidine accelerates skin wound healing in rats

O desenvolvimento de métodos que tem por objetivo acelerar e melhorar a qualidade do processo de cicatrização de feridas tem impacto positivo na condução de distúrbios de cicatrização associados a inúmeras condições médicas. Neste estudo, avaliamos os efeitos moleculares, celulares e clínicos da aplicação tópica de 5-azacitidina na cicatrização de feridas em ratos. De acordo com estudos pregressos, a 5-azacitidina reduz a expressão de folistatina, que é um regulador negativo das ativinas. Estas, por sua vez, promovem o crescimento de células em diferentes tecidos, incluindo a pele. Ratos Wistar machos com oito semanas de vida foram submetidos a um ferimento cutâneo com punch de oito milímetros na região dorsal. A seguir os ratos foram aleatoriamente separados em grupo controle (veículo) ou submetidos à aplicação tópica de 5-azacitidina (10 mM), uma vez por dia por até 12 dias, iniciando-se no terceiro dia após a lesão. A documentação fotográfica e coleta de amostras ocorreram nos dias 5, 9 e 15. O emprego desta droga resultou em aceleração da cicatrização da ferida, (99,7±7,0% versus 71,2±2,8% no dia 15, p <0,01). Este resultado clínico foi acompanhado pela redução de aproximadamente três vezes na expressão protéica de folistatina. O exame histológico da pele revelou re-epitelização eficiente com aumento da expressão de queratinócitos e aumento significativo na expressão do gene de TGF-β além da diminuição significativa de citocinas, tais como TNF-α e IL-10. Analisamos também a proliferação celular na lesão de pele através do método de incorporação de BrdU. O número de células positivas para BrdU aumentou significativamente quando comparado ao controle. No entanto, quando folistatina exógena foi aplicada na pele em paralelo ao tratamento tópico de 5-azacitidina a maioria dos benefícios do medicamento foi perdida.

The development of new methods aimed at improving wound healing may have an impact on the outcomes of a number of medical conditions. Here we evaluate the molecular and clinical effects of topical 5-azacytidine, a compound used in myelodysplasia, on the wound healing in rats. According to previous studies, 5-Azacytidine decreases the expression of follistatin 1, which is a negative regulator of activins. Activins, in turn, promote cell growth in different tissues, including the skin. Eight-week old male Wistar rats were submitted to an 8 mm punchwound in the dorsal region. After three days, rats were randomly assigned to either control or topical application of a solution containing 5-azacytidine (10mM), once a day. Photo documentation and collection of samples occurred at days 5, 9 and 15. Overall, 5-azacytidine resulted on a significant acceleration of complete wound healing (99.7% ±0.7.0 vs. 71.2%±2.8 on days 15; n=10; p<0,01). This was accompanied by an up to 3-fold reduction in follistatin expression. Histological examination of the skin revealed efficient reepithelization with increase in gene expression of TGF-β and keratinocytes markers, involucrin and citokeratin, besides the significant decrease of cytokines such as TNF-α and IL-10. In addition, we analyzed cell proliferation in injured skin employing the BrdU incorporation method. The treatment with 5-azacytidine led to a progressive increase of BrdU positive cells. Finally when recombinant follistatin was employed in the skin in parallel to topical 5-azacytidine most of the benefits of the drug were lost. Thus, 5-azacytidine acts, at least in part, through the follistatin/activin pathway to improve wound healing in rats. This study belongs to online research process Caring in Nursing and Health.

Diferenciação de células-tronco mesenquimais derivadas do tecido adiposo em cardiomiócitos / Differentiation of adipose tissue-derived mesenchymal stem cells into cardiomyocytes

FUNDAMENTO: O potencial de renovação e proliferação dos cardiomiócitos, in vivo, é pequeno, e por isso, o músculo cardíaco apresenta limitada capacidade de repor células perdidas. Na tentativa de minimizar os danos oriundos de lesões hipóxico-isquêmicas e daquelas que acometem o sistema de condução do coração, a terapia celular com células-tronco mesenquimais (MSC) vem sendo utilizada, inclusive com cardiomiócitos diferenciados a partir de MSC. OBJETIVO: O presente trabalho comparou três protocolos distintos de indução de diferenciação objetivando a sugestão de um método viável para a diferenciação de maior número de células funcionais que expressem fenótipo cardiomiogênico. MÉTODOS: Culturas de MSC obtidas de tecido adiposo de ratos jovens da linhagem Lewis transgênicos para proteína verde fluorescente (GFP) foram submetidos a três diferentes meios de diferenciação cardiogênica: Planat-Bérnard, 5-azacitidina e meio Planat-Bérnard + 5-azacitidina e observadas quanto a expressão de marcadores celulares cardíacos. RESULTADOS: Nos três protolocos utilizados observou-se formação da proteína alfa-actinina sarcomérica no citoesqueleto das células submetidas à diferenciação, expressão de conexina 43 na membrana nuclear e citoplasmática e formação de gap junctions, necessárias para a propagação do impulso elétrico no miocárdio, contudo, em nenhum protocolo foi observada contração espontânea das células submetidas à diferenciação cardiogênica. CONCLUSÃO: A indução com 5-azacitidina proporcionou diferenciação celular cadiomiogênica efetiva e similar à encontrada com o meio Planat-Bénard e, por ser um protocolo mais simples, rápido e com menor custo torna-se o método de eleição.

BACKGROUND: Cardiomyocytes have small potential for renovation and proliferation in vivo. Consequently, the heart muscle has limited capacity of self-renewal. Mesenchymal stem cells (MSC) therapy, as well as MSC differentiated into cardiomyocytes, has been used in the attempt to minimize the effects of ischemic-hypoxic lesions and those affecting the electrical conduction system of the heart. OBJECTIVE: The present study compared three distinct protocols for induced differentiation of MSC into cardiomyocytes aimed at finding a viable method for producing a large number of functional cells expressing cardiomyogenic phenotype. METHODS: Mesenchymal stem cells were obtained from the adipose tissue of young transgenic Lewis rats expressing green fluorescent protein (GFP), and submitted to three distinct differentiation-inducing media: 1) Planat-Bérnard, 2) 5-azacytidine, and 3) Planat-Bérnard + 5-azacytidine; further, these cells were identified based on the expression of cardiac cell markers. RESULTS: All three protocols detected the expression of sarcomeric-alpha-actinin protein in the exoskeleton of cells, expression of connexin-43 in the nuclear and cytoplasmic membrane, and formation of gap junctions, which are necessary for electrical impulse propagation in the myocardium. However, no spontaneous cell contraction was observed with any of the tested protocols. CONCLUSION: Induction with 5-azacytidine provided an effective cadiomyogenic cellular differentiation similar to that obtained with Planat-Bénard media. Therefore, 5-azacytidine was the method of choice for being the simplest, fastest and lowest-cost protocol for cell differentiation.

Epigenetic therapy in allogeneic hematopoietic stem cell transplantation

DNA methylation and other epigenetic phenomena appear to be relevant in the pathogenesis of several malignant disorders. DNA methyltransferases add methyl groups to cytosine-phosphate-guanine (CpG) islandsleading to gene promoter silencing. The DNA methyltransferases inhibitors azacitidine and decitabine have anti-tumor activity against a broad range of malignancies, but have been investigated mostly in myelodysplastic syndrome. In addition, these agents have immunomodulatory effects that are under investigation in the allogeneic stem cell transplantation scenario. Both drugs have been used in the perioperative period of allogeneic transplantations with varying degrees of success. It has been hypothesized that low dose azacitidine may increase the graftversus-leukemia effect and have a role in the maintenance of remission after allogeneic transplantation for myeloid leukemias. It is also intriguing that this favorable effect might occur while mitigating graft-versus-host disease. Here we present a review of the rapidly growing field of epigenetic manipulation using hypomethylating agents in allogeneic transplantation.

Cytogenetic effect of 5-azacytidine in patients with hematological malignancies

BACKGROUND: Recently, the importance of cytogenetics has grown in the diagnosis, prognosis and treatment of leukemias and myelodysplastic syndromes. 5-azacytidine is a drug that has well-known cytogenetical effects and is approved in the treatment of myelodysplastic syndromes. To date, no studies have been performed to evaluate the impact of 5-azacytidine on the chromosomes of patients with hematological neoplasias. This study aimed to investigate the effects of 5-azacytidine on chromosomes of patients with different hematological malignancies using G-band analyses to identify possible cytogenetical alterations. METHODS: The peripheral blood of 18 patients with hematological malignancies and 18 controls was collected in heparinized tubes. 5-azacytidine was added, at a final concentration of 10-5M, to cultures 7 hours prior to harvest. RESULTS: Uncoiled centromeric/pericentromeric heterochromatin of chromosomes-1, 9 and 16 occurred more frequently in the patients than in controls. This higher frequency of uncoiled heterochromatin was statistically significant (p-value = 0.004) for chromosome-9. Conversely, we observed that the fragile site at 19q13 was more frequent in controls (p-value = 0.0468). CONCLUSIONS: The results of this study suggest that satellite sequences, located in the heterochromatin of chromosome-9, are hypomethylated in hematological malignancies. This hypomethylation may contribute to the disease, activating transposable elements and/or promoting genomic instability, enabling the loss of heterozygosity of important tumor suppressor genes. An investigation of the 19q13 region may help to understand whether or not the predominant occurrence of the fragile site at 19q13 in controls is due to hypermethylation of this region.

La telomerasa: una enzima con múltiples aplicaciones en la investigación del cáncer / Telomerase: an enzyme with multiple applications in cancer research

The telomerase is a ribonucleoprotein enzyme to which multiple functions have been attributed, the most important of these is the maintenance of the telomere which is related with cellular immortalization and cancer. 85 of human tumors have telomerase activity, that in normal cells goes undetected. These characteristics make the telomerase an attractive target for chemotherapy.

Genetic and cytological characterization of a developmental mutant of Aspergillus nidulans induced by 5-azacytidine

An analysis of a new medusa mutant of Aspergillus nidulans obtained by 5-azacytidine-treatment and named B116 is provided. The B116 mutant was phenotypically characterized by the production of conidiophores with reduced pigmentation and vesicles bearing multiple tiers of sterigmata. A single nuclear gene located on chromosome I is responsible for phenotypical changes in the mutant. The 5-azacytidine-altered locus, designated medA102, is recessive in heterozygous diploid and the medusa mutant is a Dp(II,I) duplication bearer that renders the strain mitotically unstable.