Diagnostic screening of paroxysmal nocturnal hemoglobinuria: Prospective multicentric evaluation of the current medical indications.

BACKGROUND: Although consensus guidelines have been proposed in 2010 for the diagnostic screening of paroxysmal nocturnal hemoglobinuria (PNH) by flow cytometry (FCM), so far no study has investigated the efficiency of such medical indications in multicentric vs. reference laboratory settings. METHODS: Here we evaluate the efficiency of consensus medical indications for PNH testing in 3,938 peripheral blood samples submitted to FCM testing in 24 laboratories in Spain and one reference center in Brazil. RESULTS: Overall, diagnostic screening based on consensus medical indications was highly efficient (14% of PNH+ samples) both in the multicenter setting in Spain (10%) and the reference laboratory in Brazil (16%). The highest frequency of PNH+ cases was observed among patients screened because of bone marrow (BM) failure syndrome (33%), particularly among those with aplastic anemia (AA; 45%) and to a less extent also a myelodysplastic syndrome (MDS; 10%). Among the other individuals studied, the most efficient medical indications for PNH screening included: hemolytic anemia (19%), hemoglobinuria (48%) and unexplained cytopenias (9%). In contrast, only a minor fraction of the patients who had been submitted for PNH testing because of unexplained thrombosis in the absence of cytopenia, were positive (0.4%). CONCLUSIONS: In summary, our results demonstrate that the current medical indications for PNH screening by FCM are highly efficient, although improved screening algorithms are needed for patients presenting with thrombosis and normal blood cell counts. © 2016 International Clinical Cytometry Society.

Positive Impact of Eculizumab Therapy on Surgery for Budd-Chiari Syndrome in a Patient with Paroxysmal Nocturnal Hemoglobinuria and a Long-Term History of Thrombosis.

Paroxysmal nocturnal hemoglobinuria (PNH) is associated with severe end-organ damage and a high risk of thrombosis. Budd-Chiari syndrome, which develops after thrombotic occlusion of major hepatic blood vessels, is relatively common in PNH and has been associated with increased mortality. We report the case of a 46-year-old male with PNH who presented with Budd-Chiari syndrome associated with portal cavernoma, portal hypertension and hypersplenism. In September 2010, the patient suffered gastrointestinal bleeding, hematuria, and elevated plasma lactate dehydrogenase; he started eculizumab therapy with a good response. In October 2012, he developed upper gastrointestinal variceal bleeding and a splenorenal shunt was placed. At the time of writing, the patient remains stable and eculizumab continues to be effective. There is limited data on the use of eculizumab for prevention of hemolysis and its consequences in PNH patients undergoing surgery. Our findings provide evidence for the efficacy and safety of eculizumab in this setting.

Immunomodulatory effects of the intake of fermented milk with Lactobacillus casei DN114001 in lactating mothers and their children.

The healthy action of probiotics is not only due to their nutritional properties and their influence on the gastrointestinal environment, but also to their action on the immune system. The aim of the present study was to determine if 6 weeks of probiotic intake would be able to modulate the immune system in women who had recently delivered and were breast-feeding. The design consisted of a randomised, controlled and double-blind nutritional intervention study with parallel groups with a sample size of 104 women. The main variable is the T helper type 1/T helper type 2 (Th1/Th2) profile determined by measuring interferon-gamma (Th1) and IL-4 (Th2) values in peripheral blood by flow cytometry. The modifications of cytokines were evaluated in maternal milk by cytometric bead array in a flow cytometer and ELISA at three stages of breast-feeding: colostrum, early milk (10 d) and mature milk (45 d). Additionally, the anthropometry and infectious and allergic episodes in the newborn were followed up throughout the first 6 months of life. After the consumption of milk fermented with Lactobacillus casei during the puerperium, we observed a nonsignificant increase in T and B lymphocytes and a significant increase in natural killer cells. A decrease in the pro-inflammatory cytokine TNF-alpha in maternal milk and fewer gastrointestinal disturbances were also observed in the breast-fed child of the mothers who consumed L. casei. The intake of milk fermented with L. casei during the lactation period modestly contributes to the modulation of the mother's immunological response after delivery and decreases the incidence of gastrointestinal episodes in the breast-fed child.

Antiapoptotic proteins Bcl2 and BclX do not protect chronic myeloid leukemia cells from imatinib-mediated growth arrest

El imatinib (Glivec, Gleevec, STI571) es un inhibidor de la quinasa Bcr-Abl, y es el fármaco de más uso en leucemia mieloide crónica (LMC). El imatinib induce apoptosis en varias líneas celulares derivadas de LMC, entre ellas K562. Sin embargo, para obtener remisión hematológica es necesario el tratamiento continuado con imatinib, un hecho no consistente con un mecanismo de acción citotóxico in vivo del fármaco in vivo. En este trabajo hemos analizado un los efectos del imatinib en la proliferación y apoptosis de líneas celulares derivadas de K562 con expresión constitutiva de las proteínas antiapoptóticas Bcl2 y BclX. Hemos encontrado que la apoptosis mediada por imatinib era completamente abolida en las líneas celulares con expresión de Bcl2 y BclX. Sin embargo, el imatinib inhibía la proliferación, aunque este efecto fue menos severo que en las células parentales K562. Concluimos que, además de su efecto apoptótico, el imatinib actúa a través de un mecanismo independiente de la apoptosis para detener la proliferación

Imatinib (Glivec, Gleevec, STI571), a Bcr-Abl kinase inhibitor, is the most used drug in chronic myeloid leukemia. Imatinib induces apoptosis in a number of CML-derived cell lines, including K562. However, in order to achieve hematological remissions it is required chronic treatment with the drug, a fact inconsistent with a cytotoxic mechanism of imatinib in vivo. In this work we have analysed the effects of imatinib on the proliferation and apoptosis of K562-derived cell lines with constitutive expression of the anti-apoptotic genes Bcl2 and BclX. We found that imatinib-mediated apoptosis was completely abrogated in both Bcl2- and BclXcell lines. However, imatinib inhibited proliferation, although growth rate was higher than in parental K562. We conclude that, besides its apoptotic effect, imatinib acts through an apoptosis-independent mechanism to arrest cell growth

Renin expression in hematological malignancies and its role in the regulation of hematopoiesis.

It has been demonstrated that some myeloid blasts express renin, but normal bone marrow (BM) does not display this expression. The aim of the present work was to analyze the renin expression in different hematological malignancies and different myeloid cell lines. We investigated the expression of renin by RT-PCR in BM from patients with hematological malignancies (106 patients), in nine normal BM from healthy donors and in leukemic cell lines (K562, KU812, MEG-01, U-937 and HL60), as well in K562 cell line subjected to differentiation treatments. We have observed renin expression in cells from acute myeloid leukemia (AML), chronic myelogenous leukemia (CML) and acute lymphoblastic leukemia (ALL) cases. The highest frequency was observed in AML-non acute promyelocytic leukemia(APL) cases (47.2% of the cases). The disappearance of this expression was associated with the status of complete remission of AML. Renin is expressed in some myeloid human leukemia cell lines such as K562, KU812 and MEG-01. However, when K562 cells were treated with inducers of growth inhibition and/or differentiation, the expression did not disappear, indicating that renin expression is associated with a blastic phenotype rather than with cell proliferation. The obtained findings suggest that the renin expression could have a role on the disease development and could be used as an aberrant marker of leukemia.