Prolonged venous bleeding due to traditional treatment with leech bite: a case report

<p>Abstract</p> <p>Introduction</p> <p>The medicinal leech, <it>Hirudo medicinalis</it>, has been used in the treatment of many diseases for thousands of years. In Turkey, it is used most commonly in the management of venous diseases of lower extremities.</p> <p>Case presentation</p> <p>A 25-year-old Turkish woman presented to our emergency room with bleeding from her left leg. She had been treated for varicose veins in her lower extremities with leeches about 24 hours before admission to the emergency room. The bleeding was controlled by applying pressure with sterile gauze upon the wound, and she was discharged. She returned after four hours having started bleeding again. Hemostasis was achieved by vein ligation under local anesthesia.</p> <p>Conclusions</p> <p>Leech bite should be evaluated as a special injury. Prolonged bleeding can be seen after leech bites. In such cases, hemostasis either with local pressure or ligation of the bleeding vessel is mandatory.</p

Nitric Oxide And Hypoxia Response In Pluripotent Stem Cells

The expansion of pluripotent cells (ESCs and iPSCs) under conditions that maintain their pluripotency is necessary to implement a cell therapy program. Previously, we have described that low nitric oxide (NO) donor diethylenetriamine/nitric oxide adduct (DETA-NO) added to the culture medium, promote the expansion of these cell types. The molecular mechanisms are not yet known. We present evidences that ESC and iPSCs in normoxia in presence of low NO triggers a similar response to hypoxia, thus maintaining the pluripotency. We have studied the stability of HIF-1α (Hypoxia Inducible Factor) in presence of low NO. Because of the close relationship between hypoxia, metabolism, mitochondrial function and pluripotency we have analyzed by q RT-PCR the expression of genes involved in the glucose metabolism such as: HK2, LDHA and PDK1; besides other HIF-1α target gene. We further analyzed the expression of genes involved in mitochondrial biogenesis such as PGC1α, TFAM and NRF1 and we have observed that low NO maintains the same pattern of expression that in hypoxia. The study of the mitochondrial membrane potential using Mito-Tracker dye showed that NO decrease the mitochondrial function. We will analyze other metabolic parameters, to determinate if low NO regulates mitochondrial function and mimics Hypoxia Response. The knowledge of the role of NO in the Hypoxia Response and the mechanism that helps to maintain self-renewal in pluripotent cells in normoxia, can help to the design of culture media where NO could be optimal for stem cell expansion in the performance of future cell therapies

Protección de la Heparina a las células B- pancreáticas frente a radicales libres de Oxígeno.

La diabetes autoinmune tipo 1 se caracteriza por la invasión de células mononucleares en los islotes pancreáticos, destruyendo así las células beta productoras de insulina. Se ha visto que in vivo la destrucción autoinmune de los islotes se asocia con la producción de heparanasa, la cual degrada heparán sulfato (molécula imprescindible para la supervivencia de los islotes) y se permite la entrada de las células inmunitarias que atacarán los islotes beta pancreáticos. Se han obtenido resultados mediante la adición de concentraciones conocidas de heparina, la cual confiere una protección extra frente a radicales libres de oxígeno y como consecuencia hay una disminución de la mortalidad celular.A través de tres líneas celulares distintas se ha llevado a cabo el experimento. Primeramente con Rinm5F productora de insulina y somatostatina, con células Ins capaces de responder al estímulo de glucosa produciendo y secretando insulina y finalmente con fibroblastos. El experimento se basa en dejar crecer las cepas celulares en un número determinado de flacs según el tratamiento. Se añade heparina a una concentración conocida y establecida anteriormente y al día siguiente con una concentración exacta de agua oxigenada (aporta los radicales libres de oxígeno) se la añadimos al cultivo. Recogemos las células y gracias al ioduro de propidio con concentración 1 mg/ml marcamos las células muertas para así poder comprobar el porcentaje de supervivencia celular que le ha conferido la heparina a cada línea celular. Procedemos a realizar el contaje con el citómetro que indica el % de muerte celular, ya que el ioduro de propidio es un agente intercalante que se une a los ácidos nucleicos. Esta molécula fluorescente se utiliza para evaluar la viabilidad celular o el contenido de ADN en las células. Se puede utilizar para diferenciar células necróticas, apoptóticas o vivas.Los resultados obtenidos con las líneas celulares Rinm5F y las Ins nos muestran que a bajas concentraciones de agua oxigenada y con heparina, efectivamente hay protección ya que la muerte celular se reduce de un 10 a un 15%. En cambio con los fibroblastos no vemos protección a ninguna de las concentraciones establecidas, resultado que ya esperábamos en nuestra hipótesis inicial. Estos resultados sólo son el inicio de un largo estudio, ya que  primero se ajustan las concentraciones a trabajar con radicales libres de oxígeno, pero en un  futuro el estudio se realizará también con radicales de nitrógeno

Prolonged survival of patients with angioimmunoblastic T-cell lymphoma after high-dose chemotherapy and autologous stem cell transplantation: the GELTAMO experience

Abstract OBJECTIVES: Angioimmunoblastic T-cell lymphoma (AIL) is a rare lymphoma with a poor prognosis and no standard treatment. Here, we report our experiences with 19 patients treated with high-dose chemotherapy and autologous stem cell transplantation (HDC/ASCT) within the GELTAMO co-operative group between 1992 and 2004. METHODS: The median age at transplantation was 46 yr. Fifteen patients underwent the procedure as front-line therapy and four patients as salvage therapy. Most patients received peripheral stem cells (90%) coupled with BEAM or BEAC as conditioning regimen (79%). RESULTS: A 79% of patients achieved complete response, 5% partial response and 16% failed the procedure. After a median follow-up of 25 months, eight patients died (seven of progressive disease and secondary neoplasia), while actuarial overall survival and progression-free survival at 3 yr was 60% and 55%. Prognostic factors associated with a poor outcome included bone marrow involvement, transplantation in refractory disease state, attributing more than one factor of the age-adjusted-International Prognostic Index, Pretransplant peripheral T-cell lymphoma (PTCL) Score or Prognostic Index for PTCL. CONCLUSIONS: More than half of the patients with AIL that display unfavourable prognostic factors at diagnosis or relapse would be expected to be alive and disease-free after 3 yr when treated with HDC/ASCT. Patients who are transplanted in a refractory disease state do not benefit from this procedure

Relación entre muerte celular y mantenimiento de la pluripotencialidad de células mES en protocolos de diferenciación con óxido nítrico

En este proyecto se pretende estudiar la relación entre la muerte celular y la diferenciación durante el desarrollo usando como modelo experimental células madre embrionarias de ratón (mESC). Se ha descrito que las altas concentraciones de DETA-NO (500uM) provocan la diferenciación de estas mESC hacia endodermo (Mora-Castilla et al., 2010), pero en estos protocolos no todas consiguen sobrevivir y un alto porcentaje de células muere. Es por ello que se quiere analizar la relación entre ambos eventos. Se estudiará, por un lado, si el bloqueo de la apoptosis con inhibidores de caspasas tiene efectos sobre la diferenciación; y por otro lado si es probable que exista una liberación de moléculas al medio por parte de las células apoptóticas que contribuya a la pérdida de la pluripotencialidad de sus vecinas. Para ello se ha analizado el mantenimiento del estado pluripotente en dos tipos de ensayos. Por un lado se ha estudiado el efecto de inhibidores de caspasas en tratamientos con DETA-NO en dos líneas de mESC (D3 y R1/E), y por otro lado se ha evaluado el efecto del reciclaje de medios de cultivo en la línea celular D3. Para medir el mantenimiento de la pluripotencialidad se han empleado distintas técnicas de biología molecular (extracción de RNA, PCR, qPCR, Western Blotting…) y técnicas de microscopía. Con ello se han buscado diferencias de expresión de los marcadores de pluripotencia (Nanog y Oct4) y marcadores de endodermo (Pdx1, Cxcr4), mesodermo (Brachyury) y ectodermo (Zic1). Se ha demostrado que el uso de inhibidores de caspasas en tratamientos con DETA-NO bloquea la regulación a la baja de Nanog, tanto en niveles de RNAm como de proteína, y disminuye la expresión de los marcadores de diferenciación. Por otro lado, el uso de medios de cultivo reciclados procedentes de tratamientos anteriores con y sin DETA-NO, suplementados y con el factor inhibidor de la diferenciación LIF (Leukemia Inhibitory Factor) estimula la diferenciación de las células. Los resultados obtenidos apuntan a que podría existir una relación entre la muerte celular y la diferenciación, ya que al inhibir la muerte por apoptosis se favorece el mantenimiento del estado pluripotente . Además, el uso de medios reciclados ayuda a la diferenciación de las células incluso en presencia de LIF. Por ello se cree que células apoptóticas podrían estar secretando sustancias al medio que las células vecinas estarían utilizando como señales para diferenciarse

Characteristics and outcome of adult patients with acute promyelocytic leukemia and increased body mass index treated with the PETHEMA Protocols

Objective The obesity/overweight may have an influence on APL outcomes. Methods This is the biggest multicentre analysis on 1320 APL patients treated with AIDA-induction and risk-adapted consolidation between 1996 and 2012. Patients body mass index (BMI) was classified as underweight (= 30 kg/m(2)) according to the World Health Organization (WHO) criteria. Results and conclusions Relationship between male gender, older age, and other known laboratory abnormalities in overweight/obese patients was significant. The induction mortality rate was significantly higher in APL with BMI >= 25 vs BMI = 25 had a trend to lower OS (74% vs 80%; P = .06). However, in the multivariate analysis, BMI did not retain the independent predictive value (P = .46). There was no higher incidence of differentiation syndrome with BMI >= 25, but there was a trend in obese. There was no difference in relapse rate according to the BMI. In summary, overweight/obesity does not represent an independent risk factor for APL outcomes. The influence of obesity in APL patients treated with chemotherapy-free regimens remains to be established

A precision medicine test predicts clinical response after idarubicin and cytarabine induction therapy in AML patients

Complete remission (CR) after induction therapy is the first treatment goal in acute myeloid leukemia (AML) patients and has prognostic impact. Our purpose is to determine the correlation between the observed CR/CRi rate after idarubicin (IDA) and cytarabine (CYT) 3 + 7 induction and the leukemic chemosensitivity measured by an ex vivo test of drug activity. Bone marrow samples from adult patients with newly diagnosed AML were included in this study. Whole bone marrow samples were incubated for 48 h in well plates containing IDA, CYT, or their combination. Pharmacological response parameters were estimated using population pharmacodynamic models. Patients attaining a CR/CRi with up to two induction cycles of 3 + 7 were classified as responders and the remaining as resistant. A total of 123 patients fulfilled the inclusion criteria and were evaluable for correlation analyses. The strongest clinical predictors were the area under the curve of the concentration response curves of CYT and IDA. The overall accuracy achieved using MaxSpSe criteria to define positivity was 81%, predicting better responder (93%) than non-responder patients (60%). The ex vivo test provides better yet similar information than cytogenetics, but can be provided before treatment representing a valuable in-time addition. After validation in an external cohort, this novel ex vivo test could be useful to select AML patients for 3 + 7 regimen vs. alternative schedules