MOZ-TIF2, but not BCR-ABL, confers properties of leukemic stem cells to committed murine hematopoietic progenitors.

To better understand the origin of leukemic stem cells, we tested the hypothesis that all leukemia oncogenes could transform committed myeloid progenitor cells lacking the capacity for self-renewal, as has recently been reported for MLL-ENL. Flow-sorted populations of common myeloid progenitors and granulocyte-monocyte progenitors were transduced with the oncogenes MOZ-TIF2 and BCR-ABL, respectively. MOZ-TIF2-transduced progenitors could be serially replated in methylcellulose cultures and continuously propagated in liquid culture, and resulted in an acute myeloid leukemia in vivo that could be serially transplanted. In contrast, BCR-ABL transduction conferred none of these properties to hematopoietic progenitors. These data demonstrate that some, but not all, leukemia oncogenes can confer properties of leukemic stem cells to hematopoietic progenitors destined to undergo apoptotic cell death.

CT53518, a novel selective FLT3 antagonist for the treatment of acute myelogenous leukemia (AML).

Up to 30% of acute myelogenous leukemia (AML) patients harbor an activating internal tandem duplication (ITD) within the juxtamembrane domain of the FLT3 receptor, suggesting that it may be a target for kinase inhibitor therapy. For this purpose we have developed CT53518, a potent antagonist that inhibits FLT3, platelet-derived growth factor receptor (PDGFR), and c-Kit (IC(50) approximately 200 nM), while other tyrosine or serine/threonine kinases were not significantly inhibited. In Ba/F3 cells expressing different FLT3-ITD mutants, CT53518 inhibited IL-3-independent cell growth and FLT3-ITD autophosphorylation with an IC(50) of 10-100 nM. In human FLT3-ITD-positive AML cell lines, CT53518 induced apoptosis and inhibited FLT3-ITD phosphorylation, cellular proliferation, and signaling through the MAP kinase and PI3 kinase pathways. Therapeutic efficacy of CT53518 was demonstrated both in a nude mouse model and in a murine bone marrow transplant model of FLT3-ITD-induced disease.

PKC412 overcomes resistance to imatinib in a murine model of FIP1L1-PDGFRα-induced myeloproliferative disease.

FIP1L1-PDGFRalpha causes hypereosinophilic syndrome (HES) and is inhibited by the tyrosine kinase inhibitor imatinib (Gleevec). Imatinib is a potent inhibitor of ABL, ARG, PDGFRalpha, PDGFRbeta, and KIT and induces durable hematologic responses in HES patients. However, we observed relapse with resistance to imatinib as consequence of a T674I mutation in FIP1L1-PDGFRalpha, analogous to the imatinib-resistant T315I mutation in BCR-ABL. We developed a murine bone marrow transplant model of FIP1L1-PDGFRalpha-induced myeloproliferative disease to evaluate the efficacy of PKC412, an alternative inhibitor of PDGFRalpha, for the treatment of HES. PKC412 is effective for treatment of FIP1L1-PDGFRalpha-induced disease and of imatinib-induced resistance due to the T674I mutation. Our data establish PKC412 as molecularly targeted therapy for HES and other diseases expressing activated PDGFRalpha and demonstrate the potential of alternative kinase inhibitors to overcome resistance in target tyrosine kinases.

Plant extracts with anti-inflammatory properties--a new approach for characterization of their bioactive compounds and establishment of structure-antioxidant activity relationships.

Geranium robertianum L. (Geraniacea) and Uncaria tomentosa (Willd.) DC. (Rubiaceae) plant extracts, frequently used in traditional medicine for treatment of inflammatory and cancer diseases, were studied to identify potential bioactive compounds that may justify their therapeutic use and their underlying mechanisms of action. Since some of the pharmacological properties of these plant extracts may be linked to their antioxidant potential, the antioxidant activity, in relation to free radical scavenging, was measured by the ABTS/HRP and DPPH() assays, presenting U. tomentosa the higher activity. The antioxidant activity was also evaluated by scavenging of HOCl, the major strong oxidant produced by neutrophils and a potent pro-inflammatory agent. U. tomentosa was found to be a better protector against HOCl, which may justify its effectiveness against inflammatory diseases. SPE/LC-DAD was used for separation/purification purposes and ESI-MS/MS for identification/characterization of the major non-volatile components, mainly flavonoids and phenolic acids. The ESI-MS/MS methodology proposed can be used as a model procedure for identification/characterization of unknowns without the prerequisite for standard compounds analysis. The ESI-MS/MS data obtained were consistent with the antioxidant activity results and structure-activity relationships for the compounds identified were discussed.

Nurses- puerperal mothers interaction: searching for cultural care.

OBJECTIVE: To identify the importance of the cultural care dimension in the nurse-puerperal mother interaction. METHOD: Qualitative study based on content analysis. To collect data, 36 semi structured interviews were applied (21 nurses and 15 women who had recently given birth). The participants' free and informed consent was ensured and all ethical requirements were respected. Data analysis was supported by NVivo 10 software. RESULTS: The following categories were considered the most relevant: (i) nurses' attitudes that puerperal mothers consider crucial in health care provision; (ii) nurses' cultural competence; (iii) language resources used by nurses in their interaction with people from other cultures; (iv) the limitations that were observed during nurses-new mothers interactions. CONCLUSION: Evidence shows that, generally speaking, nurses claim to provide personalized health care assistance. However data revealed that the kind of interaction observed between nurses and women who had just given birth has not always been the most effective and that the former often show they lacked the appropriate cultural competence to deal with the latter's expectations and needs.

FLT3 internal tandem duplication mutations induce myeloproliferative or lymphoid disease in a transgenic mouse model.

Activating FMS-like tyrosine kinase 3 (FLT3) mutations have been identified in approximately 30% of patients with acute myelogenous leukemia (AML), and recently in a smaller subset of patients with acute lymphoblastic leukemia (ALL). To explore the in vivo consequences of an activating FLT3 internal tandem duplication mutation (FLT3-ITD), we created a transgenic mouse model in which FLT3-ITD was expressed under the control of the vav hematopoietic promoter. Five independent lines of vav-FLT3-ITD transgenic mice developed a myeloproliferative disease with high penetrance and a disease latency of 6-12 months. The phenotype was characterized by splenomegaly, megakaryocytic hyperplasia, and marked thrombocythemia, but without leukocytosis, polycythemia, or marrow fibrosis, displaying features reminiscent of the human disease essential thrombocythemia (ET). Clonal immature B- or T-lymphoid disease was observed in two additional founder mice, respectively, that could be secondarily transplanted to recipient mice that rapidly developed lymphoid disease. Treatment of these mice with the FLT3 tyrosine kinase inhibitor, PKC412, resulted in suppression of disease and a statistically significant prolongation of survival. These results demonstrate that FLT3-ITD is capable of inducing myeloproliferative as well as lymphoid disease, and indicate that small-molecule tyrosine kinase inhibitors may be an effective treatment for lymphoid malignancies in humans that are associated with activating mutations in FLT3.

Nurses- puerperal mothers interaction: searching for cultural care / Interacción enfermeras y madres: en la búsqueda de un cuidado cultural / Interação enfermeiros e puérperas: na procura de um cuidado cultural

ABSTRACT Objective: To identify the importance of the cultural care dimension in the nurse-puerperal mother interaction. Method: Qualitative study based on content analysis. To collect data, 36 semi structured interviews were applied (21 nurses and 15 women who had recently given birth). The participants' free and informed consent was ensured and all ethical requirements were respected. Data analysis was supported by NVivo 10 software. Results: The following categories were considered the most relevant: (i) nurses' attitudes that puerperal mothers consider crucial in health care provision; (ii) nurses' cultural competence; (iii) language resources used by nurses in their interaction with people from other cultures; (iv) the limitations that were observed during nurses-new mothers interactions. Conclusion: Evidence shows that, generally speaking, nurses claim to provide personalized health care assistance. However data revealed that the kind of interaction observed between nurses and women who had just given birth has not always been the most effective and that the former often show they lacked the appropriate cultural competence to deal with the latter's expectations and needs.

RESUMEN Objetivo: identificar la dimensión de la competencia cultural en la interacción entre enfermeras y mujeres que hayan dado a luz recientemente. Método: estudio cualitativo basado en el análisis de contenidos. En la recopilación de datos se utilizó la entrevista semiestructurada. Han participado 21 enfermeras y 15 madres en un total de 36 entrevistas. Se aseguró el consentimiento previo, libre e informado de todos los participantes y todos los requisitos éticos fueron respetados. El análisis de datos fue apoyado por el programa NVivo 10. Resultados: las siguientes categorías fueron destacadas: (i) actitudes de las enfermeras que las madres creen ser indispensables en el proceso de cuidar; ii) el ejercicio de la competencia cultural; iii) los recursos lingüísticos utilizados por las enfermeras en la interacción con personas de otras culturas; iv) restricciones y problemas observados en las interacciones entre enfermeras y madres. Conclusión: se concluye que, en general, las enfermeras afirman que cuidan de sus pacientes de manera personalizada. Todavía, los datos revelaron que las interacciones entre enfermeras y madres son a menudo poco eficaces y a las enfermeras les resulta difícil demostrar su competencia cultural.

RESUMO Objetivo: Identificar a dimensão do cuidado cultural na interação entre enfermeiras e puérperas. Método: Estudo qualitativo assente na análise de conteúdo. Na colheita de dados se utilizou a entrevista semiestruturada, a 21 enfermeiras e 15 puérperas num total de 36 entrevistadas. Se assegurou o consentimento livre e esclarecido bem como respeitaram-se todos os requisitos éticos. A análise de dados foi suportada pelo NVivo 10. Resultados: Destacaram-se as categorias: (i) Atitudes das enfermeiras que as puérperas reconhecem imprescindíveis no processo de cuidar; (ii) Exercitar a competência cultural; (iii) Recursos linguísticos utilizados pelas enfermeiras na interação com pessoas de outras culturas; (iv) Constrangimentos manifestados nas interações enfermeiras e puérperas. Conclusão: Conclui-se que apesar das enfermeiras, ao nível discursivo, afirmarem que cuidam de forma personalizada, os dados revelaram que nem sempre as enfermeiras foram eficazes nas interações com as puérperas, de forma a demonstrarem a competência cultural, num estadio considerado culturalmente consciente.

PML/RARalpha and FLT3-ITD induce an APL-like disease in a mouse model.

Acute promyelocytic leukemia (APL) cells invariably express aberrant fusion proteins involving the retinoic acid receptor alpha (RARalpha). The most common fusion partner is promyelocytic leukemia protein (PML), which is fused to RARalpha in the balanced reciprocal chromosomal translocation, t(15;17)(q22:q11). Expression of PML/RARalpha from the cathepsin G promoter in transgenic mice causes a nonfatal myeloproliferative syndrome in all mice; about 15% go on to develop APL after a long latent period, suggesting that additional mutations are required for the development of APL. A candidate target gene for a second mutation is FLT3, because it is mutated in approximately 40% of human APL cases. Activating mutations in FLT3, including internal tandem duplication (ITD) in the juxtamembrane domain, transform hematopoietic cell lines to factor independent growth. FLT3-ITDs also induce a myeloproliferative disease in a murine bone marrow transplant model, but are not sufficient to cause AML. Here, we test the hypothesis that PML/RARalpha can cooperate with FLT3-ITD to induce an APL-like disease in the mouse. Retroviral transduction of FLT3-ITD into bone marrow cells obtained from PML/RARalpha transgenic mice results in a short latency APL-like disease with complete penetrance. This disease resembles the APL-like disease that occurs with long latency in the PML/RARalpha transgenics, suggesting that activating mutations in FLT3 can functionally substitute for the additional mutations that occur during mouse APL progression. The leukemia is transplantable to secondary recipients and is ATRA responsive. These observations document cooperation between PML/RARalpha and FLT3-ITD in development of the murine APL phenotype.