Two death pathways induced by sorafenib in myeloma cells: puma-mediated apoptosis and necroptosis

Purpose. Sorafenib is a multikinase inhibitor that targets the MAPK pathway and is currently used for the treatment of hepatocellular and renal carcinoma. Recently, it has been shown that sorafenib is also cytotoxic to multiple myeloma (MM) cells. Here, we have further analyzed the mechanism of sorafenib-induced death in MM cells. Methods. Cell death induced by sorafenib in MM cell lines and in plasma cells from MM patients was evaluated by analysis of gene expression by RT-MLPA and quantitative PCR, protein levels and functionality by Western blot and flow cytometry and gene silencing with siRNA. Results. Cell death was characterized by phosphatidylserine exposure, ΔΨm loss, cytochrome c release and caspase activation, hallmarks of apoptosis. DL50 at 24 h ranged from 6 to 10 µM. Ex vivo treatment with 20 µM sorafenib induced apoptosis in around 80 % myeloma cells from six multiple myeloma patients. Sorafenib induced caspase-dependent degradation of Bcl-xL and Mcl-1 proteins, destabilizing the mitochondria and speeding up the development of apoptosis. Sorafenib treatment increased levels of Puma at mRNA and protein level and gene silencing with siRNA confirmed a relevant role for Puma in the induction of apoptosis. Co-treatment with the pan-caspase inhibitor Z-VAD-fmk prevented cell death to a variable degree depending on the cell line. In RPMI 8226 cells, Z-VAD-fmk prevented most of sorafenib-induced death. However, death in MM.1S was only prevented by co-incubation with both Z-VAD-fmk and the RIP1K inhibitor necrostatin-1, indicating that under conditions of inefficient caspase activation, sorafenib induces death by necroptosis. Conclusion. Our results demonstrate a key role for Puma in the triggering of sorafenib-induced apoptosis and that this drug can also induce death by necroptosis in multiple myeloma cells (AU)

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Two death pathways induced by sorafenib in myeloma cells: Puma-mediated apoptosis and necroptosis.

PURPOSE: Sorafenib is a multikinase inhibitor that targets the MAPK pathway and is currently used for the treatment of hepatocellular and renal carcinoma. Recently, it has been shown that sorafenib is also cytotoxic to multiple myeloma (MM) cells. Here, we have further analyzed the mechanism of sorafenib-induced death in MM cells. METHODS: Cell death induced by sorafenib in MM cell lines and in plasma cells from MM patients was evaluated by analysis of gene expression by RT-MLPA and quantitative PCR, protein levels and functionality by Western blot and flow cytometry and gene silencing with siRNA. RESULTS: Cell death was characterized by phosphatidylserine exposure, ΔΨm loss, cytochrome c release and caspase activation, hallmarks of apoptosis. DL50 at 24 h ranged from 6 to 10 µM. Ex vivo treatment with 20 µM sorafenib induced apoptosis in around 80 % myeloma cells from six multiple myeloma patients. Sorafenib induced caspase-dependent degradation of Bcl-xL and Mcl-1 proteins, destabilizing the mitochondria and speeding up the development of apoptosis. Sorafenib treatment increased levels of Puma at mRNA and protein level and gene silencing with siRNA confirmed a relevant role for Puma in the induction of apoptosis. Co-treatment with the pan-caspase inhibitor Z-VAD-fmk prevented cell death to a variable degree depending on the cell line. In RPMI 8226 cells, Z-VAD-fmk prevented most of sorafenib-induced death. However, death in MM.1S was only prevented by co-incubation with both Z-VAD-fmk and the RIP1K inhibitor necrostatin-1, indicating that under conditions of inefficient caspase activation, sorafenib induces death by necroptosis. CONCLUSION: Our results demonstrate a key role for Puma in the triggering of sorafenib-induced apoptosis and that this drug can also induce death by necroptosis in multiple myeloma cells.

Efectos hemodinámicos de dos gases anestésicos distintos sobre las fases anhepática y de reperfusión del transplante hepático / Hemodynamic effects of two different anesthetic agents during anhepatic and reperfusion stages of liver transplantation

OBJETIVOS: La enfermedad hepática crónica en fases terminales se caracteriza por un estado hiperdinámico. Hemos analizado los efectos hemodinámicos del sevoflurano y desflurano durante las fases anhepática y de reperfusión del trasplante hepático. PACIENTES Y METODO: Estudiamos 58 pacientes sometidos a trasplante a los cuales se les administró como anestésico de mantenimiento sevoflurano o desflurane. Las determinaciones hemodinámicas fueron: basal (postinducción) (A), 5 minutos antes de pinzas la porta (B), 10 minutos después de pinzar la cava (C1), 5 minutos antes de revascularizar (C), 5 minutos tras revascularizar el injerto (D1) y a los 30 minutos de despinzar (D2). Los parámetros hemodinámicos analizados fueron los siguientes: frecuencia cardíaca (FC), presión arterial media (PAM), presión venosa central (PVC), presión de la arteria pulmonar media (PAPM), presión capilar pulmonar (PCP), gasto cardíaco (GC), índice cardíaco (IC), índices de resistencias vasculares sistémicas y pulmonares (IRVS, IRVP), transporte de oxígeno (DO2), consumo de oxígeno (VO2) y saturación en sangre venosa mixta (SVO2). RESULTADOS: La FC, PAM; PVC, PAPM, PCP, DO2 y SVO2 las diferencias no fueron significativas desde el punto de vista estadístico entre ambos gases anestésicos. El GC y el IC presentaron valores más altos en todo momento con el desflurano que con el sevoflurano, siendo las diferencias estadísticamente significativas (p < 0,05) en B. C2; D1. Los IRVS siguieron una tendencia descendente desde el comienzo de la intervención. En todo momento el sevoflurano mantuvo unos valores más elevados que el desflurano, siendo las diferencias muy significativas (p < 0,01) en B y C2, en D2 significativas (p < 0,05). Con respecto a los IRVP, con el sevoflurano se mantuvieron más elevados. cuyas diferencias fueron significativas (p < 0,05) en B. En el grupo del desflurano, el V02 tras revascularizar el injerto, tendió a elevarse y sus diferencias fueron significativas (p < 0,05) a los 5 minutos de la fase neohepática. CONCLUSIONES: En nuestra experiencia, el desflurano mantiene un GC y un IC con valores más elevados que el sevoflurano, antes de la fase anhepática que permanecen tras la revascularización del injerto. Ello puede ser la consecuencia de que los IRVS son menores con el desflurano, produciendo, por tanto, mayor vasodilatación periférica en la fase anhepática. Sin embargo, no existen variaciones entre ambos anestésicos respecto a la SVO2 lo que nos hace sospechar que con un buen manejo anestésico no existirían muchas diferencias prácticas entre los dos grupos (AU)

Successful lipid-complexed amphotericin B treatment of Candida arthritis in a lymphoma patient.

Fungal arthritis is uncommon but has been increasingly diagnosed over recent years, particularly in patients with immunodeficiency due for instance to hematological malignancies. Candida albicans is the most frequent causative agent, and the knee is the joint most often involved. Amphotericin B is the drug of choice, but is associated with significant toxicity. Recently developed lipid formulations of amphotericin B have been found as effective and less toxic than the conventional formulation. We report a new case of Candida arthritis that occurred after chemotherapy for nonHodgkin's lymphoma and was successfully treated with lipid-complexed amphotericin B.

Hemorheological profile in chronic venous insufficiency after surgery.

In recent years, there have been rheological abnormalities reported in chronic venous insufficiency (CVI), mainly an increase of erythrocyte aggregability (EA), which probably take part in the pathophysiology of the disease. The aim of this study was to analyze the hemorheological profile after stripping in 45 patients suffering from CVI. Follow-up included laboratory tests on the 7th, 60th and 180th day after surgery. EA was assessed with a photometric aggregometer (MA1, Myrenne) in stasis and low shear (3 s(-1)). The results show an increase of EA on the 7th day after surgery (p<0.001). Two and 6 months later, EA values returned to those found prior to surgery. The plasma fibrinogen level changes in a way parallel to EA. The association between rheological disturbances and thrombogenesis is well known, so the hyperaggregability found supports the antithrombotic prophylaxis in the early postsurgical period. On the other hand, the hemorheological abnormalities persist after stripping, so postsurgical treatment to inhibit EA may be beneficial.

[Hypercoagulability status previous to total hip and knee arthroplasty: the contribution of rheumatoid arthritis]. / Estado hipercoagulativo previo a la artroplastia total de cadera y rodilla: contribución de la artritis reumatoide.

PURPOSE: Starting from a status hypercoagulability previous to substitutive hip and knee surgery, the aim of this work was to investigate the influence of different osteoarthropatic pictures for which arthroplasty is indicated in the activation of the clotting cascade, rheumatoid arthritis (RA) being one of such pictures. PATIENTS AND METHODS: Of 79 patients suitable for prosthetic surgery of hip (53) and knee (26), the preoperative values of several markers, namely, D dimers (D-D), thrombin-antithrombin (TAT) complex, and F1 + 2 prothrombin fragment (F1 + F2) were assessed by enzymoimmunoasay. The mean age of the patients was 65.5 years, and their sex distribution was 50 women and 29 men. The indications for arthroplasty were as follows: osteoarthrosis (62), aseptic necrosis (11), RA (9), articular gout (2), previous fracture (2), more than one diagnosis overlapped in some cases. The results attained were compared with a control group comprised of 33 subjects (16 women and 17 men) with mean age similar to the patient's group (68.06 years). RESULTS: The D-D values in the patients suitable for hip arthroplasty and the TAT values in patients suitable for both types of surgery were significantly higher than those found in the control group (p = 0.012 and 0.01, respectively). The preoperative TAT levels of the RA patients were significantly higher (p = 0.025) than those found in the patients with the other surgical indications. CONCLUSIONS: Previously to the performance of arthroplasty, the patients show hypercoagulative marker values higher than those of age-matched controls. The significant rising of TAT found in RA patients is concordant with the literature, and this fact makes it advisable to include RA among the pathologic situations associated with hypercoagulability, as this is a common indication for substitutive hip and knee surgery with high risk of venous thromboembolic disease.

Fatal myelofibrosis following fludarabine administration in a patient with indolent lymphoma.

We report a case of fulminant myelofibrosis after administration of fludarabine in a patient diagnosed as having refractory low-grade lymphoma, progressing fatally. Myelofibrosis in the setting of an indolent lymphoma is very rare; this fact, and the short period between drug administration and fibrosis suggest an etiopathogenic link, although this potential and severe adverse effect of fludarabine has not been previously reported in the literature.

Incidence and clinical manifestations of activated protein C resistance and factor V Leiden in young patients with venous thromboembolic disease in Spain.

In order to evaluate the actual incidence and clinical repercussion of activated protein C resistance (APCR) in our area, we performed a coagulation and thrombophillic study on 65 young patients diagnosed with deep vein thrombosis and 53 controls. Family and genetic study was carried out in APC-resistant patients. We found APCR in 26.15% of patients and the 77.7% of these and their relative were heterozygous for factor V Leiden. There's a clear relationship between phenotype APCR and thrombosis, and also between factor V Leiden and thrombosis.

[Comparative study of 2 antithrombotic regimens in 75 patients with coronary endoprostheses]. / Estudio comparativo de dos pautas antitrombóticas en 75 pacientes portadores de endoprótesis coronaria.

PURPOSE: To compare systemic anticoagulation with antiaggregation in patients with coronary stent, with regard to subacute occlusion, mean hospital staying and haemorrhagic complications. PATIENTS AND METHODS: Seventy-five patients with coronary stent were treated with one of two different antithrombotic protocols. A group comprised of 34 patients (group A) received sodium heparin and acenocoumarin, plus acetylsalicylic acid (325 mg) and dipyridamole (225 mg). The remaining 41 patients (group B) were given antiplatelet agents, namely ticlopidine (125-250 mg) and aspirin (125 mg). RESULTS: One case of group A (2.9%) showed thrombosis due to stent occlusion. No thrombotic complications were seen in the patients with antiplatelet drugs. Haemorrhagic complications were present in 11 group A patients (32.3%), and blood transfusion was necessary in 3 of them. Hemorrhage was present in 9 cases of group B (21.8%), and none of them needed blood transfusion. The mean number of days to achieve INR > 2 was 3.06 (1-11) in group and 2.02 (1-5) in group B. CONCLUSIONS: Antiplatelet regimes appear as a good choice in coronary stent, in spite of the fact that the primary indication seems that of group A.