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Evaluation of shRNA-mediated gene silencing by electroporation in LPB fibrosarcoma cells

References Rols MP, Delteil C, Golzio M, Dumond P, Cros S, Teissie J. In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol 1998; 16(2): 168-71. Wells JM, Li LH, Sen A, Jahreis GP, Hui SW. Electroporation-enhanced gene delivery in mammary tumors. Gene Ther 2000; 7(7): 541-7. Bettan M, Ivanov MA, Mir LM, Boissiere F, Delaere P, Scherman D. Efficient DNA electrotransfer into tumors. Bioelectrochemistry 2000; 52(1): 83

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Expansive growth of two glioblastoma stem-like cell lines is mediated by bFGF and not by EGF

Abstract

Background. Patient-derived glioblastoma (GBM) stem-like cells (GSCs) represent a valuable model for basic and therapeutic research. GSCs are usually propagated in serum-free Neural Basal medium supplemented with bFGF and EGF. Yet, the exact influence of these growth factors on GSCs is still unclear. Recently it was suggested that GBM stemlike cells with amplified EGFR should be cultured in stem cell medium without EGF, as the presence of EGF induced rapid loss of EGFR amplification. However, patient biopsies are usually taken into culture before their genomic profiles are defined. Thus, an important question remains whether GBM cells without EGFR amplification also can be cultured in stem cell medium without EGF.

Meterials and methods. To address this question, we used two heterogeneous glioblastoma GSC lines (NCH421k and NCH644) that lack EGFR amplification.

Results. Although both cell lines showed very low EGFR expression under standard growth conditions, bFGF stimulation induced higher expression of EGFR in NCH644. In both cell lines, expression of the stem cell markers nestin and CD133 was higher upon stimulation with bFGF compared to EGF. Importantly, bFGF stimulated the growth of both cell lines, whereas EGF had no effect. We verified that the growth stimulation by bFGF was either mediated by proliferation (NCH421k) or resistance to apoptosis (NCH644).

Conclusions. We demonstrate that GSC cultures without EGFR amplification can be maintained and expanded with bFGF, while the addition of EGF has no significant effect and therefore can be omitted.

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Atrial Fibrillation and Acute Myocardial Infarction – An Inflammation-Mediated Association

ABSTRACT

Atrial fibrillation (AF) is an increasingly widespread healthcare problem. AF can frequently present as a complication in acute coronary syndromes (ACS), especially in ST-elevation acute myocardial infarction (AMI), in which case it is the most frequent supraventricular rhythm disturbance with an estimated incidence of 6.8-21%. The presence of AF in ACS heralds worse outcomes in comparison to subjects in sinus rhythm, and several studies have shown that in AMI patients, both new-onset and pre-existing AF are associated with a higher risk of major adverse cardiovascular and cerebrovascular events during hospitalization. The cause of newonset AF in AMI is multifactorial. Although still incompletely understood, the mechanisms involved in the development of AF in acute myocardial ischemic events include the neurohormonal activation of the sympathetic nervous system that accompanies the AMI, ischemic involvement of the atrial myocytes, ventricular dysfunction, and atrial overload. The identification of patients at risk for AF is of great significance as it may lead to prompt therapeutic interventions and closer follow-up, thus improving prognosis and decreasing cardiovascular and cerebrovascular events. The present manuscript aims to summarize the current research findings related to new-onset AF in AMI patients, as well as the predictors and prognostic impact of this comorbid association.

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Focused transhepatic electroporation mediated by hypersaline infusion through the portal vein in rat model. Preliminary results on differential conductivity

Abstract

Background

Spread hepatic tumours are not suitable for treatment either by surgery or conventional ablation methods. The aim of this study was to evaluate feasibility and safety of selectively increasing the healthy hepatic conductivity by the hypersaline infusion (HI) through the portal vein. We hypothesize this will allow simultaneous safe treatment of all nodules by irreversible electroporation (IRE) when applied in a transhepatic fashion.

Material and methods

Sprague Dawley (Group A, n = 10) and Athymic rats with implanted hepatic tumour (Group B, n = 8) were employed. HI was performed (NaCl 20%, 3.8 mL/Kg) by trans-splenic puncture. Deionized serum (40 mL/Kg) and furosemide (2 mL/Kg) were simultaneously infused through the jugular vein to compensate hypernatremia. Changes in conductivity were monitored in the hepatic and tumour tissue. The period in which hepatic conductivity was higher than tumour conductivity was defined as the therapeutic window (TW). Animals were monitored during 1-month follow-up. The animals were sacrificed and selective samples were used for histological analysis.

Results

The overall survival rate was 82.4% after the HI protocol. The mean maximum hepatic conductivity after HI was 2.7 and 3.5 times higher than the baseline value, in group A and B, respectively. The mean maximum hepatic conductivity after HI was 1.4 times higher than tumour tissue in group B creating a TW to implement selective IRE.

Conclusions

HI through the portal vein is safe when the hypersaline overload is compensated with deionized serum and it may provide a TW for focused IRE treatment on tumour nodules.

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Testing of mechanisms of action of rituximab and clinical results in high-risk patients with aggressive CD20+ lymphoma

monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement mediated lysis. Blood 2000; 95 : 3900-8. Winkler U, Jensen M, Manzke O, Schulz H, Diehl V, Engert A. Cytokine-release syndrome in patients with B-cell chronic lymphocytic leukemia and high lymphocyte counts after treatment with an anti-CD20 monoclonal antibody (rituximab, IDEC-C2B8). Blood 1999; 94 : 2217-24. Shipp MA, Harrington DP, Anderson JR, Armitage JO, Bonadonna G, Brittinger G, et al. A predictive model for aggressive non

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Electrochemotherapy by pulsed electromagnetic field treatment (PEMF) in mouse melanoma B16F10 in vivo

clinic, was chosen in the study to test the application of induced electroporation mediated with magnetic field. The stock solution of the chemotherapeutic drug used in the study, CDDP (5 mg/mL, Cysplatyl, Aventis Laboratory, Paris, France) was dissolved in aqua pro injection and frozen in aliquots of 1 mL. In order that each animal received a dose of 80 μg of CDDP, a fresh solution at appropriate concentration of CDDP (1 mg/mL) was prepared in 0.9% sodium chloride solution daily before each experiment. Mouse tumor model Female C57B1/6 mice were purchased from

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p38 MAPK regulates the expression of ether à go-go potassium channel in human osteosarcoma cells

Prever A,Vassallo E, et al. High-dose chemotherapy in the treatment of relapsed osteosarcoma: an Italian sarcoma group study. J Clin Oncol 2002; 20: 2150-6. 19. Marina N, Gorlick R. Immune approaches to treating osteosarcoma. CancerBiol Ther 2009; 8: 981-3. 20. He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B. A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 1998; 95: 2509-14. 21. Gao YS, Mei J, Tong TL, Hu M, Xue HM, Cai XS. Inhibitory effects of VEGFsiRNA mediated

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Coronary Plaque Geometry and Thoracic Fat Distribution in Patients with Acute Chest Pain – a CT Angiography Study

Cardiovasc Imaging. 2010;3:1104-1112. doi: 10.1016/j.jcmg.2010.07.014. 8. Mazurek T, Zhang L, Zalewski A, et al. Human epicardial adipose tissue is a source of inflammatory mediators. Circulation. 2003;108:2460-2466. doi: 10.1161/01.CIR.0000099542.57313.C5. 9. Mahabadi AA, Massaro JM, Rosito GA, et al. Association of pericardial fat, intrathoracic fat, and visceral abdominal fat with cardiovascular disease burden: the Framingham Heart Study. Eur Heart J. 2009;30:850-856. doi: 10.1093/eurheartj/ehn573. 10. Ding J, Hsu FC, Harris TB, et al. The association

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Lucifer Yellow uptake by CHO cells exposed to magnetic and electric pulses

conductivity on electropermeabilization and survival of cells in vitro. Bioelectrochemistry 2001; 54 : 107-15. Kotnik T, Pucihar G, Miklavčič D. Induced transmembrane voltage and its correlation with electroporation-mediated molecular transport. J Membrane Biol 2010; 236 : 3-13. Pucihar G, Krmelj J, Reberšek M, Batista Napotnik T, Miklavčič D. Equivalent pulse parameters for electroporation. IEEE T Biomed Eng 2011; 58 : 3279-88. Rols MP, Femenia P, Teissié J. Long-lived macropinocytosis

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Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

genotoxins. Mutat Res-Gen Tox En 2008; 653 : 63-9. Zhou B-BS, Elledge SJ. The DNA damage response: putting checkpoints in perspective. Nature 2000; 408 : 433-9. Sionov RV, Haupt Y. The cellular response to p53: the decision between life and death. Oncogene 1999; 18 : 6145-57. Waldman T, Kinzler KW, Vogelstein B. P21 is necessary for the P53-mediated G1 arrest in human cancer cells. Cancer Res 1995; 55 : 5187-90. Vogelstein B, Lane D, Levine AJ. Surfing the p

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