Distribution by Scientific Domains
Distribution within Medical Sciences

Terms modified by MTT

  • mtt method
  • mtt reduction

  • Selected Abstracts

    Bifunctional Eu3+ -doped Gd2O3 nanoparticles as a luminescent and T1 contrast agent for stem cell labeling

    Zhilong Shi
    Abstract Magnetic resonance tracking of stem cells has recently become an emerging application for investigating cell,tissue interactions and guiding the development of effective stem cell therapies for regeneration of damaged tissues and organs. In this work, anionic Eu3+ -doped Gd2O3 hybrid nanoparticles were applied as a contrast agent both for fluorescence microscopy and T1 -weighted MRI. The nanoparticles were synthesized through the polyol method and further modified with citric acid to obtain anionic nanoparticles. These nanoparticles were internalized into human mesenchymal stem cells (hMSCs) as confirmed by confocal laser scanning microscopy and quantified by inductively coupled plasma,mass spectrometry. MTT assay of the labeled cells showed that the nanoparticles did not possess significant cytotoxicity. In addition, the osteogenic, adipogenic and chondrogenic differentiation of the hMSCs was not influenced by the labeling process. With MRI, the in vitro detection threshold of cells after incubation with nanoparticles at a Gd concentration of 0.5,mMfor 2,h was estimated to be about 10 000 cells. The results from this study indicate that the biocompatible anionic Gd2O3 nanoparticles doped with Eu3+ show promise both as a luminescent and T1 contrast agent for use in visualizing hMSCs. Copyright © 2010 John Wiley & Sons, Ltd. [source]

    Effect of temperature and storage media on human periodontal ligament fibroblast viability

    Beatriz Dulcineia Mendes Souza
    The purpose of this study was to compare the effectiveness of several storage media to preserve cultured periodontal ligament fibroblasts (PDLF) under different temperatures. The media tested were: sterile Hank's balanced salt solution (sHBSS), non-sterile HBSS (nHBSS), skimmed milk, Save-A-Tooth®, Minimum Essential Medium (MEM) and water (negative control). MEM at 37°C was used as positive control. PDLF were obtained from explants of extracted healthy human teeth. Plates containing confluent PDLF were soaked in the various media for 3, 6, 24, 48 and 72 h at 37°C and 20°C. After incubation, viability of the cells was determined using the tetrazolium salt-based colorimetric (MTT) assay and the Trypan Blue exclusion test after 6, 24, 48 and 72 h of incubation at 20°C. The results were analyzed statistically using Kruskal,Wallis, Scheffé and Mann,Whitney (, = 5%) tests. Results from the MTT assay at 37°C and 20°C showed that skimmed milk was the best storage medium for up to 24 and 48 h, respectively, followed by nHBSS and sHBSS. Results from the Trypan Blue exclusion test showed that the best storage media were milk, sHBSS and nHBSS, with no statistical differences, for any time period. The Save-A-Tooth® had a detrimental effect on cells after 24 h. The influence of temperature on the effectiveness of the storage media tested showed at 20°C a decreasing order of efficacy as follows: milk > sHBSS and nHBSS > MEM > Save-A-Tooth® > water while at 37°C it was: MEM > nHBSS > milk > sHBSS > Save-A-Tooth® > water. In conclusion, incubation temperature altered the effectiveness of the storage media and skimmed milk at 20°C was better than HBSS in maintaining PDLF viability. [source]

    Synthesis and screening of substituted 1,4-naphthoquinones (NPQs) as antifilarial agents

    Nisha Mathew
    Abstract Eleven amino-substituted 1,4-naphthoquinones were synthesized via the reaction of 1,4-naphthoquinone with different primary and secondary mono- and diamines in the presence of dichloromethane ethanol (1:2) solvent at room temperature. All compounds were purified by flash column chromatography, characterized by TLC, HPLC, 13C-NMR, 1H-NMR, and FT-IR spectral analysis and were evaluated in vitro for antifilarial activity using adult bovine filarial worm Setaria digitata by assessing worm motility and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction. Seven of the 11 compounds had macrofilaricidal activity with compounds 9 (2-[(1,3-dimethylbutyl) amino] naphthalene-1,4-dione) and 11 (2-(4-methylpiperazin-1-yl) naphthalene-1,4-dione) having maximum activity (ED50 values of 0.91 and 1.2,µM, respectively, at 48,h). The effect of different substitutions on antifilarial activity is discussed. Drug Dev Res 2009. © 2009 Wiley-Liss, Inc. [source]

    Enhanced sensitivity to DNA damage induced by cooking oil fumes in human OGG1 deficient cells

    Mei Wu
    Abstract Cooking oil fumes (COFs) have been implicated as an important nonsmoking risk factor of lung cancer in Chinese women. However, the molecular mechanism of COFs-induced carcinogenicity remains unknown. To understand the molecular basis underlying COFs-induced cytotoxicity and genotoxicity as well as the roles of hOGG1 in the repair of COFs-induced DNA damage, a human lung cancer cell line with hOGG1 deficiency, A549-R was established by using a ribozyme gene targeting technique that specifically knockdowned hOGG1 in A549 lung adenocarcinoma cells. MTT and comet assays were employed to examine cell viability and DNA damage/repair, respectively, in A549-R and A549 cell lines treated with COF condensate (COFC). RT-PCR and Western blot results showed that the expression of hOGG1 in A549-R cell line was significantly decreased compared with that in A549 cell line. The concentration of COFC that inhibited cell growth by 50% (the IC50) in the A549-R cell line was much lower than that in the A549 cell line, and more COFC-induced DNA damage was detected in the A549-R cell line. The time course study of DNA repair demonstrated delayed repair kinetics in the A549-R cell line, suggesting a decreased cellular damage repair capacity. Our results showed that hOGG1 deficiency enhanced cellular sensitivity to DNA damage caused by COFC. The results further indicate that hOGG1 plays an important role in repairing COF-induced DNA damage. Our study suggests that COFs may lead to DNA damage that is subjected to hOGG1 -mediated repair pathways, and oxidative DNA damage may be involved in COF-induced carcinogenesis. Environ. Mol. Mutagen., 2008. © 2008 Wiley-Liss, Inc. [source]

    Cytotoxicity assessment of gliotoxin and penicillic acid in Tetrahymena pyriformis

    C. Gräbsch
    Abstract Various studies have documented the associations between mold exposure and effects on health. Mycotoxins, which occur in spores and mold fragments, can be involved in processes that have pathological effects, such as adynamia of the immune system, recurrent infections of the respiratory tract, or asthma. Using Tetrahymena pyriformis, a single-cell organism well established as a suitable model for human respiratory epithelium-cell functionalities, we investigated dose,response relationships of the mycotoxins gliotoxin and penicillic acid. Our study focused on the viability (cell count, MTT assay), energy levels (adenosine-5,-triphosphate content), energy-providing processes (MTT reduction per cell), and cell respiration (oxygen consumption). Both mycotoxins acted as cytotoxins in a dose-dependent manner. Gliotoxin had a stronger inhibitory effect (EC50 0.38 ,M) than did penicillic acid (EC50 343.19 ,M). The energy-providing processes were not inhibited or were only weakly inhibited under the influence of gliotoxin, whereas penicillic acid caused stimulation of the physiological parameters. Summarizing the results, it is clear that the two investigated mycotoxins must have different modes of action. They are not only different in the strength of their toxic effects but also in a variety of physiological aspects. In addition, T. pyriformis showed differences in its ability to overcome the negative effects of particular mycotoxin exposures. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 111,117, 2006. [source]

    Characteristics of okadaic acid,induced cytotoxic effects in CHO K1 cells

    C. Huynh-Delerme
    Abstract This article reports the results of investigations into the process of cell death induced in the Chinese hamster ovary cell K1 subclone (CHO K1) by okadaic acid (OA), a hydrophobic polyether produced by marine dinoflagellates. The IC50 was about 13 nM OA after 24 h of treatment, as determined using neutral red. With the MTT assay, the IC50 was 25 nM, although in this case 25% of the initial staining was still observed at 100 nM. Hoechst staining showed that mitotic figures accumulated at 12 nM OA after a 24- or 48-h treatment. In experiments limited to a 3-day treatment without changing the medium, CHO K1 cells were engaged in the death process at 50 nM OA after about 20 h and at 10 nM OA after 48 h. In many cells nuclear fragmentation that resulted in the apparent appearance of vesicles correlated with increasing cellular volume. But additional cell fragmentation was not observed with any treatment, and the chromatin material seemed to progressively disappear inside the cells. DNA fragmentation was analyzed by electrophoresis and with the TUNEL technique. With both techniques, the DNA was fragmented by 48 h in both 25 and 50 nM OA. Electrophoresis showed that both adherent and nonadherent cells were affected. Annexin-positive/ propidium iodide (PI),negative cells were rarely observed after OA treatment. Some were seen under the scanning cytometer after 20 h at 50 nM OA or after 48 h at 10 nM OA, but they were never detected by flow cytometry. Most of the time scanning cytometry showed either unstained cells or PI-positive (annexin-positive or -negative) cells (48 h, 50 nM, or 72 h, 10 nM). Flow cytometry cytograms showed two cell subpopulations: one composed of a majority of smaller cells, the other of larger cells. The larger cells markedly decreased with time and OA treatment (50 and 100 nM). Stained-cell counting showed that all cells that stained were both annexin- and PI positive and that most PI-positive cells were smaller. Ki67 antigen labeling showed the proliferative activity of CHO K1 cultures but also demonstrated the loss of this activity in smaller cells treated with 50 nM OA for 48 h. We concluded that in our culture conditions the main OA target within CHO K1 cultures was dividing cells. Our results suggest that cells with disturbed metaphase,anaphase enter apoptosis, leading to necrotic daughter cells. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 383,394, 2003 [source]

    Anthracene photoinduced toxicity to plhc-1 cell line (Poeciliopsis lucida) and the role of lipid peroxidation in toxicity

    Jonghoon Choi
    Abstract Many polycyclic aromatic hydrocarbons (PAHs) are acutely toxic to fish and other aquatic organisms in the presence of solar ultraviolet radiation (SUVR) of environmentally realistic intensities. In the present study, the photoinduced toxicity of a PAH (anthracene; ANT) to topminnow hepatoma cell line (PLHC-1) was assessed. After the toxicity was characterized, the role of lipid peroxidation in PAH photoinduced toxicity was examined by measuring lipid peroxidation products and by assessing the effect of lipid peroxidation antagonist (Trolox) treatment. In cytotoxicity tests using two assays (MTT, neutral red), the SUVR/ANT treatment elicited toxicity to PLHC-1 cells in a concentration- and SUVR (exposure duration and intensity)-dependent pattern. As found in previous organism-level studies, no significant cytotoxicity was observed in the cells exposed either to fluorescent light/ANT or to SUVR only. The SUVR/ANT treatment elicited the lipid peroxidation process and Trolox pretreatment significantly reduced SUVR/ANT-induced cell mortality. Microscopic observation showed that Trolox pretreatment relieved the SUVR/ANT-inflicted damage, such as cell shrinkage and membrane disruption. Together with a recent finding in our lab that increased production of superoxide anion and a lipid peroxidation product (malondialdehyde) was found in SUVR/ANT-treated fish microsomes, the present study suggests that reactive oxygen radical-induced lipid peroxidation is an important factor in PAH photoinduced toxicity to fish. [source]

    Cytotoxicity of settling particulate matter and sediments of the Neckar River (Germany) during a winter flood

    Henner Hollert
    Abstract To investigate the cytotoxic and genotoxic potentials of settling particulate matter (SPM) carried by the Neckar River, a well-studied model for a lock-regulated river in central Europe, during a flood, acute cytotoxicity was investigated using the fibroblast-like fish cell line RTG-2 with the neutral red retention, the succinic acid dehydrogenase (MTT), and the lactatedehydro-genase (LDH) release assays as well as microscopic inspection as endpoints. Genotoxicity of water, pore water, sediments, and SPM were assessed using the Ames test. Different extraction methods (Soxhlet extraction with solvents of variable polarity as well as a fluid/fluid extraction according to pH) in addition to a supplementation of biotests with S9 fractions from the liver of ,-naphthoflavone/phenobarbital-induced rats allowed a further characterization of the biological damage. Both sediments and SPM extracts caused cytotoxic effects in RTG-2 cells. Cytotoxicity was found to increase significantly with polarity of extracting solvents (NR50 = effective concentration for 50% cell death in the neutral red test: 80 [65], 100 [70], 180 [220], and 225 [270] mg/ml for ethanol, acetone, dichloromethane, and n -hexane extracts, respectively, if measured with [without] S9 supplementation). Following extraction according to pH, cytotoxicity could be attributed mainly to neutral substances (NR50: 80 and 218 mg dry SPM/ml test medium for the neutral and the acid fractions, respectively), whereas the slightly acid and basic fractions already showed little or no cytotoxicity. Samples taken during the period of flood rise showed the highest cytotoxic activities. Cytotoxicity was significantly enhanced by the addition of S9 preparations. In contrast, no genotoxic activity was found in native surface waters, pore waters, and SPM. [source]

    Iron enhances endothelial cell activation in response to Cytomegalovirus or Chlamydia pneumoniae infection

    A. E. R. Kartikasari
    Abstract Background, Chronic inflammation has been implemented in the pathogenesis of inflammatory diseases like atherosclerosis. Several pathogens like Chlamydia pneumoniae (Cp) and cytomegalovirus (CMV) result in inflammation and thereby are potentially artherogenic. Those infections could trigger endothelial activation, the starting point of the atherogenic inflammatory cascade. Considering the role of iron in a wide range of infection processes, the presence of iron may complicate infection-mediated endothelial activation. Materials and methods, Endothelial intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and endothelial selectin (E-selectin) expression were measured using flow cytometry, as an indication of endothelial activation. Cytotoxicity was monitored using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Immunostaining was applied to measure Cp and CMV infectivity to endothelial cells. Results, An increased number of infected endothelial cells in a monolayer population leads to a raised expression of adhesion molecules of the whole cell population, suggesting paracrine interactions. Iron additively up-regulated Cp-induced VCAM-1 expression, whereas synergistically potentiated Cp-induced ICAM-1 expression. Together with CMV, iron also enhanced ICAM-1 and VCAM-1 expression. These iron effects were observed without modulation of the initial infectivity of both microorganisms. Moreover, the effects of iron could be reversed by intracellular iron chelation or radical scavenging, conforming modulating effects of iron on endothelial activation after infections. Conclusions, Endothelial response towards chronic infections depends on intracellular iron levels. Iron status in populations positive for Cp or CMV infections should be considered as a potential determinant for the development of atherosclerosis. [source]

    Activation of p53 signalling in acetylsalicylic acid-induced apoptosis in OC2 human oral cancer cells

    C.-C. Ho
    Abstract Background, Nonsteroidal anti-inflammatory drugs (NSAIDs) such as acetylsalicylic acid (ASA, aspirin) are well known chemotherapeutic agents of cancers; however, the signalling molecules involved remain unclear. The aim of this study was to investigate the possible existence of a putative p53-dependent pathway underlying the ASA-induced apoptosis in OC2 cells, a human oral cancer cell line. Materials and methods, The methyl tetrazolium (MTT) assay was employed to quantify differences in cell viability. DNA ladder formation on agarose electrophoresis was used as apoptosis assay. The expression levels of several master regulatory molecules controlling various signal pathways were monitored using the immunoblotting techniques. Flow cytometry was used to confirm the effect of ASA on cell cycle. Patterns of changes in expression were scanned and analyzed using the NIH image 1·56 software (NIH, Bethesda, MD, USA). All the data were analyzed by anova. Results, Acetylsalicylic acid reduced cell viability and presence of internucleosomal DNA fragmentation. In the meanwhile, phosphorylation of p53 at serine 15, accumulation of p53 and increased the expression of its downstream target genes, p21 and Bax induced by ASA. The expression of cyclooxygenase-2 was suppressed. Disruption of p53-murine double minute-2 (MDM2) complex formation resulted in increasing the expression of MDM2 60-kDa cleavage fragment. Inhibited the activation of p42/p44 mitogen-activated protein kinase (MAPK) by PD98059, a specific inhibitor of extracellular regulatory kinase (ERK), significantly decreased cell viability and enhanced the expression of p53 induced by ASA. The result of the cell-cycle analysis showed that ASA and PD98059 induced the cell cycle arrested at the G0/G1 phase and resulted in apoptosis. Conclusion, Nonsteroidal anti-inflammatory drug-inhibited cyclooxygenase is not the only or even the most important mechanism of inhibition. Our study presents evidences that activation of p53 signalling involved in apoptosis induced by ASA. Furthermore, the apoptotic effect was enhanced by blocking the activation of p42/p44 MAPK in response to treatment with ASA, thus indicating a negative role for p42/p44 MAPK. [source]

    Mesothelin, a possible target for immunotherapy, is expressed in primary AML cells

    Daniel Steinbach
    Abstract Background:, Mesothelin is a promising candidate for tumor-specific therapy because of its limited expression in normal tissues and high expression in several cancers. The expression of the protein mesothelin in hematological malignancies has not yet been analyzed. SS1(dsFv)PE38 is a recombinant anti-mesothelin immunotoxin which is undergoing clinical evaluation in patients with mesothelin-expressing tumors. Methods and Results:, In this study we show that the mesothelin protein is expressed in leukemic cells from children with acute myeloid leukemia (AML). This finding was confirmed by western blot, immunocytochemistry and real time polymerase chain reaction (PCR). Despite the expression of mesothelin, the patient samples were not sensitive to immunotoxin SS1(dsFv)PE38 in MTT assays. Conclusions:, Primary AML cells express mesothelin but SS1(dsFv)PE38 is not active in killing these cells. Other approaches that utilize mesothelin as a target might be more effective and should be tested against AML cells. [source]

    Synthesis, Cytotoxicity and Antibacterial Studies of p -Methoxybenzyl-Substituted and Benzyl-Substituted N-Heterocyclic Carbene,Silver Complexes

    Siddappa Patil
    Abstract p -Methoxybenzyl-substituted and benzyl-substituted N-heterocyclic carbene (NHC) [(3a,c) and (6a,c)] precursors were synthesised from the reaction of 1H -imidazole (1a), 4,5-dichloro-1H -imidazole (1b), and 1H -benzimidazole (1c) with p -methoxybenzyl bromide (2) and benzyl bromide (5). These NHC precursors were then treated with silver(I) acetate to yield the NHC,silver complexes [1,3-bis(4-methoxybenzyl)imidazol-2-ylidene]silver(I) acetate (4a), [4,5-dichloro-1,3-bis(4-methoxybenzyl)imidazol-2-ylidene]silver(I) acetate (4b), [1,3-bis(4-methoxybenzyl)benzimidazol-2-ylidene]silver(I) acetate (4c), (1,3-dibenzylimidazol-2-ylidene)silver(I) acetate (7a), (1,3-dibenzyl-4,5-dichloroimidazol-2-ylidene)silver(I) acetate (7b), and (1,3-dibenzylbenzimidazol-2-ylidene)silver(I) acetate (7c), respectively. The NHC precursor 3c, four NHC,silver complexes 4c and 7a,c were characterised by single-crystal X-ray diffraction method. The preliminary antibacterial activity of all the compounds was studied against Gram-negative bacteria Escherichia coli, and Gram-positive bacteria Staphylococcus aureus using the Kirby,Bauer disk-diffusion method. Almost all the NHC,silver complexes have shown high antibacterial activity compared to the NHC precursors. In addition, the NHC,silver complexes had their cytotoxicity investigated through MTT-based preliminary in vitro testing on the Caki-1 cell lines in order to determine their IC50 values. NHC,silver complexes 4a,c and 7a,c were found to have IC50 values of 7.3 (+/,6), 12.7(+/,3), 25.2 (+/,5), 2.5 (+/,3), 10.8 (+/,4) and 12.5 (+/,4) ,M respectively on the Caki-1 cell line. [source]

    [Tris(pyrazolyl)methane]ruthenium Complexes Capable of Inhibiting Cancer Cell Growth

    Jesse M. Walker
    Abstract The [tris(pyrazolyl)methane]ruthenium complexes [(,3 -tpm)RuCl(solv)2]PF6 [tpm = tris(pyrazolyl)methane; solv = MeCN, dmso] and [(,3 -tpm)RuCl(LL)]PF6 [LL = ,2 -dppe, ,2 -dppp, ,2 -dppb, (PMePh2)2] have been prepared, characterized and screened in vitro for their antiproliferative properties against the MCF-7 (breast) and HeLa (cervical) cancer cell lines by using the MTT assay. Although the MeCN and dmso complexes showed no activity under the conditions used, the phosphane complexes exhibited remarkable cytotoxic behaviour. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009) [source]

    Synthesis and Cytotoxicity Studies of Fluorinated Derivatives of Vanadocene Y

    Brendan Gleeson
    Abstract From the reaction of 6-(2-fluoro-4-methoxyphenyl)fulvene (1a), 6-(3-fluoro-4-methoxyphenyl)fulvene (1b) and 6-[4-(trifluoromethoxy)phenyl]fulvene (1c) with LiBEt3H, lithiated cyclopentadienide intermediates (2a,c) were synthesised. These intermediates were then transmetallated to vanadium with VCl4 to yield the benzyl-substituted vanadocenes bis[(2-fluoro-4-methoxybenzyl)cyclopentadienyl]vanadium(IV) dichloride (3a), bis[(3-fluoro-4-methoxybenzyl)cyclopentadienyl]vanadium(IV) dichloride (3b), and bis[(4-trifluoromethoxybenzyl)cyclopentadienyl]vanadium(IV) dichloride (3c). The three vanadocenes 3a,c were characterised by single-crystal X-ray diffraction. All three vanadocenes had their cytotoxicity investigated through MTT-based preliminary in-vitro testing on the LLC-PK and Caki-1 cell lines in order to determine their IC50 values. Vanadocenes 3a,c were found to have IC50 values of 6.0 (+/,4), 35 (+/,7) and 13 (+/,3) ,M on the LLC-PK cell line and IC50 values of 78 (+/,11), 18 (+/,16) and 2.2 (+/,0.5) ,M on the Caki-1 cell line respectively. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009) [source]

    In Vitro Characteristics of Surface-Modified Biphasic Calcium Phosphate/Poly(L -Lactide) Biocomposite

    Weizhong Yang
    Abstract Surface-modified biphasic calcium phosphate (BCP)/poly(L -lactide) (PLLA) biocomposite is shown to have improved microstructure and mechanical properties compared to the unmodified system. In vitro biodegradation and bioactivity of the composite are investigated in simulate body fluid for up to four weeks. Weight changes of the samples and the pH changes of the SBF are recorded. Surface properties of the composite after immersion are characterized by XRD, SEM and EDX analyses. Cyto-compatibility was determined by MTT assay with L929 mouse fibroblasts. The difference of the degradation behavior between modified BCP/PLLA and the reference unmodified composite are investigated, and mBCP/PLLA composite is proved to be a better as a scaffold material. The surface formed bio-apatite layer after immersion shows the excellent bioactivity of the mBCP/PLLA composite. L929 cells show a high growth rate and proliferation, demonstrating the good cytocompatibility of mBCP/PLLA composite. [source]

    Cytotoxicity and Cell Cycle Effects of Bare and Poly(vinyl alcohol)-Coated Iron Oxide Nanoparticles in Mouse Fibroblasts

    Morteza Mahmoudi
    Super-paramagnetic iron oxide nanoparticles (SPIONs) are recognized as powerful biocompatible materials for use in various biomedical applications, such as drug delivery, magnetic-resonance imaging, cell/protein separation, hyperthermia and transfection. This study investigates the impact of high concentrations of SPIONs on cytotoxicity and cell-cycle effects. The interactions of surface-saturated (via interactions with cell medium) bare SPIONs and those coated with poly(vinyl alcohol) (PVA) with adhesive mouse fibroblast cells (L929) are investigated using an MTT assay. The two SPION formulations are synthesized using a co-precipitation method. The bare and coated magnetic nanoparticles with passivated surfaces both result in changes in cell morphology, possibly due to clustering through their magnetostatic effect. At concentrations ranging up to 80,×,10,3,M, cells exposed to the PVA-coated nanoparticles demonstrate high cell viability without necrosis and apoptosis. In contrast, significant apoptosis is observed in cells exposed to bare SPIONs at a concentration of 80,×,10,3,M. Nanoparticle exposure (20,80,×,10,3,M) leads to variations in both apoptosis and cell cycle, possibly due to irreversible DNA damage and repair of oxidative DNA lesions, respectively. Additionally, the formation of vacuoles within the cells and granular cells indicates autophagy cell death rather than either apoptosis or necrosis. [source]

    Dehydroepiandrosterone inhibits the proliferation and induces the death of HPV-positive and HPV-negative cervical cancer cells through an androgen- and estrogen-receptor independent mechanism

    FEBS JOURNAL, Issue 19 2009
    Roma A. Girón
    Dehydroepiandrosterone (DHEA) has a protective role against epithelial-derived carcinomas; however, the mechanisms remain unknown. We determined the effect of DHEA on cell proliferation, the cell cycle and cell death in three cell lines derived from human uterine cervical cancers infected or not with human papilloma virus (HPV). We also determined whether DHEA effects are mediated by estrogen and androgen receptors. Proliferation of C33A (HPV-negative), CASKI (HPV16-positive) and HeLa (HPV18-positive) cells was evaluated by violet crystal staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction. Flow cytometry was used to evaluate the phases of the cell cycle, and cell death was detected using a commercially available carboxyfluorescein apoptosis detection kit that determines caspase activation. DNA fragmentation was determined using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Flutamide and ICI 182,780 were used to inhibit androgen and estrogen receptors, respectively, and letrozol was used to inhibit the conversion of DHEA to estradiol. Our results show that DHEA inhibited cell proliferation in a dose-dependent manner in the three cell lines; the DHEA IC50 doses were 50, 60 and 70 ,m for C33A, CASKI and HeLa cells, respectively. The antiproliferative effect was not abrogated by inhibitors of androgen and estrogen receptors or by an inhibitor of the conversion of testosterone to estradiol, and this effect was associated with an increase in necrotic cell death in HPV-negative cells and apoptosis in HPV-positive cells. These results suggest that DHEA strongly inhibits the proliferation of cervical cancer cells, but its effect is not mediated by androgen or estrogen receptor pathways. DHEA could therefore be used as an alternative in the treatment of cervical cancer. [source]

    Antitumor and antifungal activities in endophytic fungi isolated from pharmaceutical plants Taxus mairei, Cephalataxus fortunei and Torreya grandis

    Yaojian Huang
    Abstract The purpose of this work was to screen the endophytic fungi having antitumor or antifungal activity, which were isolated from the inner barks of three kinds of pharmaceutical plants, Taxus mairei, Cephalataxus fortunei and Torreya grandis, collected from Fujian province, China. Antitumor activity was studied by the MTT assay and antifungal activity was determined by observing fungal growth inhibition. 13.4% of endophytic fungi fermentation broths displayed cytotoxic activity on HL-60 cells at and below a dilution of 1:50, and 6.4% on KB cells. 52.3% of endophytic fungi fermentation broths displayed growth inhibition on at least one pathogenic fungi, such as Neurospora sp., Trichoderma sp. and Fusarium sp. Among all endophytic fungi isolated, the genus Paecilomyces sp. has the highest positive rate of antitumor and antifungal activity. These results indicate that endophytic fungi could be a promising source for antitumor and antifungal bioactive agents. [source]

    The study of cell-mediated immune response in recurrent vulvovaginal candidiasis

    U. Nawrot
    Abstract The aim of this work was to examine in vitro the ability of cells from patients with recurrent vulvovaginal candidiasis (RVVC) to cell-mediated immune response. Peripheral blood mononuclear cells (PBMC) and whole blood cells (WBC) of 37 RVVC patients in acute infection and 14 in remission were examined for the ability to proliferation and cytokines production (IFN, TNF, IL-6). As a control, a group of 25 healthy women were examined. The cells were stimulated with Candida antigen (HKCA), LPS and PHA. To indicate the level of cytokines, the following cell-lines were used: A549 for IFN, WEHI 164 for TNF and 7TD1 for IL-6. The proliferation/death of cells was determined by colorimetric test using MTT. Distinct suppression of cell-mediated immune response (CMI) was shown in all patients comparing to the control. Greatest suppression was found in the acute phase of the disease. The ability of cells to proliferate and produce IFN increases only in remission. The data seem to suggest that in this phase of disease, the ability of cell-mediated immune response is restored. It was also indicated that IFN may take part in protection against Candida infection. [source]

    The effect of modulation of , -glutamyl transpeptidase and nitric oxide synthase activity on GSH homeostasis in HepG2 cells

    Inga Kwiecie
    Abstract High glutathione (GSH) level and elevated , -glutamyl transpeptidase (,GT) activity are hallmarks of tumor cells. Toxicity of drugs and radiation to the cells is largely dependent on the level of thiols. In the present studies, we attempted to inhibit ,GT activity in human hepatoblastoma (HepG2) cells to examine whether the administration of ,GT inhibitors, acivicin (AC) and 1,2,3,4-tetrahydroisoquinoline (TIQ) influences cell proliferation and enhances cytostatic action of doxorubicin (DOX) and cisplatin (CP) on HepG2 cells. The effects of these inhibitors were determined by 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), BrdU and lactate dehydrogenase (LDH) tests and by estimation of GSH level. Additionally, we investigated the changes in caspase-3 activity, which is a marker of apoptosis. The obtained results showed that the ,GT inhibitors introduced to the medium alone elicited cytotoxic effect, which was accompanied by an increase in GSH level in the cells. TIQ concomitantly increased caspase-3 activity. Doxorubicin and CP proved to be cytotoxic, and both inhibitors augmented this effect. As well DOX as CP radically decreased GSH levels, whereas ,GT inhibitors had diverse effects. Therefore, the obtained results confirm that ,GT inhibitors can enhance pharmacological action of DOX and CP, which may permit clinicians to decrease their doses thereby alleviating side effects. Aminoguanidine (nitric oxide synthase inhibitor) given alone was little cytotoxic to HepG2 cells, while its introduction to the medium together with DOX and CP significantly increased their cytotoxicity. Aminoguanidine on its own did not show any effect on GSH level in HepG2 cells, but markedly and significantly elevated its concentration when added in combination with CP but not with DOX. This indicates that when CP was used as a cytostatic, GSH level rose after treatment with its combination with both AC and aminoguanidine. [source]

    Enhancement of neuronal outward delayed rectifier K+ current by human monocyte-derived macrophages

    GLIA, Issue 14 2009
    Dehui Hu
    Abstract Macrophages are critical cells in mediating the pathology of neurodegenerative disorders and enhancement of neuronal outward potassium (K+) current has implicated in neuronal apoptosis. To understand how activated macrophages induce neuronal dysfunction and injury, we studied the effects of lipopolysaccharide (LPS)-stimulated human monocytes-derived macrophage (MDM) on neuronal outward delayed rectifier K+ current (IK) and resultant change on neuronal viability in primary rat hippocampal neuronal culture. Bath application of LPS-stimulated MDM-conditioned media (MCM) enhanced neuronal IK in a concentration-dependentmanner, whereas non-stimulated MCM failed to alter neuronal IK. The enhancement of neuronal IK was repeated in a macrophage-neuronal co-culture system. The link of stimulated MCM (MCM(+))-associated enhancement of IK to MCM(+)-induced neuronal injury, as detected by PI/DAPI (propidium iodide/4,,6-diamidino-2-phenylindol) staining and MTT assay, was demonstrated by experimental results showing that addition of IK blocker tetraethylammonium to the culture protected hippocampal neurons from MCM(+)-associated challenge. Further investigation revealed elevated levels of Kv 1.3 and Kv 1.5 channel expression in hippocampal neurons after addition of MCM(+) to the culture. These results suggest that during brain inflammation macrophages, through their capacity of releasing bioactive molecules, induce neuronal injury by enhancing neuronal IK and that modulation of Kv channels is a new approach to neuroprotection. © 2009 Wiley-Liss, Inc. [source]

    Anticancer effect of photodynamic therapy with hexenyl ester of 5-aminolevulinic acid in oral squamous cell carcinoma

    Yeon-Hee Moon MS
    Abstract Background. Five-aminolaevulinic acid (ALA) and its derivatives act as precursors of the photosensitizer protoporphyrin IX (PpIX). In this study, the effect of photodynamic therapy (PDT) with hexenyl ester of ALA (ALA-hx) was examined in a human oral squamous cell carcinoma, YD10B cells. Methods. PpIX accumulation and mRNA expression of coproporphyrinogen oxidase (CPO) by ALA and ALA-hx was examined. Cell viability was examined by MTT assay and the molecular mechanism was investigated. Results. The PpIX synthesis and mRNA expression of CPO was much higher in the cells treated with ALA-hx than ALA. At the concentration that PDT with ALA did not affect cell growth, ALA-hx PDT effectively produced reactive oxygen species (ROS) and suppressed cell growth. Growth inhibition by ALA-hx PDT was due to mitochondrial-dependent apoptosis. Conclusion. Our results suggest that ALA-hx PDT effectively induced apoptosis of YD-10B cells and can be considered as a therapeutic alternative for oral cancer. © 2009 Wiley Periodicals, Inc. Head Neck, 2010 [source]

    Prediction of poor survival by cyclooxygenase-2 in patients with T4 nasopharyngeal cancer treated by radiation therapy: Clinical and in vitro studies

    Wen-Cheng Chen MD
    Abstract Background. This study was undertaken to determine the status of cyclooxygenase-2 (COX-2) in nasopharyngeal cancer (NPC) in Taiwanese patients and its relationship to survival after radiotherapy (RT). In addition, the effect of NS-398, a potent selective COX-2 inhibitor, was tested in vitro alone and in combination with radiation on NPC-BM1 human NPC cells as a prelude to using this drug along with RT in the treatment of patients with NPC. Methods. Thirty-seven patients diagnosed with T4N0,3M0 NPC were enrolled into this study. COX-2 expression was determined by immunohistochemical staining of formalin-fixed, paraffin-embedded tumor tissue. Patient survival was the clinical end point. The effects of COX-2 expression on cell survival and radioresistance was tested in vitro using the selective COX-2 inhibitor NS-398 in conjunction with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide (MTT) and clonogenic assays. Results. COX-2 immunoreactivity was detected in 62% of NPC tumors, and expression levels were high in 43%. Survival analysis showed the 5-year overall survival rates for patients who had high COX-2 expression was 27% compared with 60% for those with low/absent expression (p = .047). Pattern of failure analysis showed no significant difference between high and low COX-2 expression in locoregional failure (27% vs 25%, p = .91). However, patients with N0 to N1 disease and high COX-2 expression had a significantly higher incidence of distant metastasis compared with patients with stage N0 to N1 disease and low COX-2 expression (83% vs 15%, p = .004). This difference was not observed in patients with N2 to N3 disease. This difference contributed to worse survival of patients whose tumors had high COX-2 expression levels. The selective COX-2 inhibitor NS-398 was directly cytotoxic to NPC-BM1 cells in vitro, as judged in an MTT assay (viable cells decreased from 92% to 76%, 52%, and 22%, with increases of NS-398 from 20 to 40, 60, and 80 ,M, respectively). Radiation-induced cell death was also increased by treatment with NS-398. At a 10% survival level, 40 ,M NS-398 increased radiation cytotoxicity by a factor of 1.37, whereas 60 ,M increased it by a factor of 4.9. Conclusions. COX-2 overexpression is a predictor for poor survival for advanced stage NPC. In vitro, NS-398 radiosensitizes the NPC-BM1 cell line, providing a basis for testing the combination of COX-2 inhibitors with radiation in the treatment of patients with NPC. © 2005 Wiley Periodicals, Inc. Head Neck27: XXX,XXX, 2005 [source]

    Lipoxin A4 inhibited hepatocyte growth factor-induced invasion of human hepatoma cells

    Xiao-Yan Zhou
    Aim:, Inflammation is a critical component of tumor progression. Lipoxin A4 (LXA4) has been approved for potent anti-inflammatory properties. Recently, it was reported that LXA4 repressed the expression and activity of cyclooxygenase-2 (COX-2), which is essential for invasion. However, there are few reports dealing with its effects on cancer. To explore whether LXA4 regulate invasion, the effects of LXA4 and its receptor agonist BML-111 on hepatocyte growth factor (HGF)-induced invasion of hepatoma cells and the possible mechanisms were researched. Methods:, Lipoxin A4 receptor (ALX) expression in HepG2 cells were measured through reverse transcription polymerase chain reaction and western blot. Cytotoxicity of LXA4 and BML-111 to HepG2 cells was detected by MTT and (3H)-TdR incorporation assay. Cell migration and invasion assays were performed using a Boyden chemotaxis chamber. COX-2 expression was detected by real-time polymerase chain reaction and western blot, respectively. Moreover, the expressions of matrix metalloproteinases (MMP)-2, MMP-9, I,B, and nuclear factor-,B (NF-,B) p65 were observed via western blot, and NF-,B transcriptional activity was tested by transfections and luciferase activities assay. Results:, ALX expression was detected in HepG2 cells, and suitable concentrations of LXA4 and BML-111 had no cytotoxicity to cells. LXA4 and BML-111 inhibited HGF-induced migration and invasion; downregulated COX-2, MMP-2 and -9; restrained HGF-induced I,B, degradation, NF-,B translocation and the transcriptional activity of NF-,B in HepG2 cells. Furthermore, exogenous PGE2 could reverse the inhibitory effects of LXA4 also BML-111 on HGF-induced invasion and migration partially. Conclusion:, LXA4 inhibited HGF-induced invasion of HepG2 cells through NF-,B/COX-2 signaling pathway partially. [source]

    Potentiation of isoniazid-induced liver toxicity by rifampicin in a combinational therapy of antitubercular drugs (rifampicin, isoniazid and pyrazinamide) in Wistar rats: A toxicity profile study

    HEPATOLOGY RESEARCH, Issue 10 2007
    Sheikh Abdullah Tasduq
    Aim:, Biochemical characterization of long-term toxic manifestations of anti-tubercular (anti-TB) drugs , rifampicin (RIF), isoniazid (INH) and pyrazinamide (PZA) , individually and in two combinations: (i) RIF + INH, and (ii) RIF + INH + PZA in Wistar rats. Methods:, Animals received anti-TB drugs , alone or in combination , once daily p.o. for up to 90 days (doses, in mg/kg: RIF, 250; INH, 50; PZA, 100). Assays for alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin (serum) and lipid peroxidation (LPO), glutathione (GSH), glutathione peroxidase (GPx), catalase, Na+K+-ATPase and CYP 2E1 (liver) were performed to assess liver toxicity. Clinical biochemistry was done by commercial kits. Determinations were made at 0, 15, 30 and 90 days of treatment schedule. Results:, Anti-TB drugs-treated animals showed abnormal rises or falls (>1.5,2 fold) in the serum/liver parameters. Mild hyperlipidemia, hypercholesterolemia and hyperuricemia were the other pathologies. Of all the treated groups, INHalone or in combination with other drugs produced a progressive enhancement of toxicity over 15,90 days. The in vivo results were further supported by in vitro results (MTT assay, GSH and LPO) in primary cultures of rat hepatocyte. Results indicated that anti-TB drugs in combination: (i) caused membrane damage resulting in leakage of ALT, ALP and bilirubin; (ii) caused imbalance in endogenous enzymatic oxidant,antioxidant defense via increased lipid peroxidation and in glutathione homeostasis; and (iii) enhanced the CYP 2E1-mediated bioactivation mechanism. Conclusion:, Toxicity manifestations seemed to be heptocytic injury targeted at hepatocytes, bile ducts or sinusoidal cells related to hepatitis and primary biliary cholestasis. [source]

    Alpha-fetoprotein-specific transfer factors downregulate alpha-fetoprotein expression and specifically induce apoptosis in Bel7402 alpha-fetoprotein-positive hepatocarcinoma cells

    Hui Zhang
    Aim:, To investigate the mechanisms of AFP-specific transfer factors (AFP-TF) in induced Bel7402 cells apoptosis. Further, we investigate the interaction between AFP-TF and AFP in the apoptosis. Methods:, Bel7402 and HepG2 AFP-positive hepatocarcinoma cell lines, SK-Hep-1 AFP-negative hepatocarcinoma cell line and Changliver normal liver cell line are used. Cell viability is evaluated by MTT assay and apoptosis is measured by Hoechst33342 staining and TUNEL assay. FACS is used to analyze the cell cycle. AFP expression is examined by RT-PCR, Western blotting and immunocytochemistry. The interaction between AFP-TF and AFP in the apoptosis is investigated by addition of AFP in cultures or AFP transfection in Bel7402 cells prior to AFP-TF treatment. Mitochondrial membrane potential (,,m) and intracellular Ca2+ concentration are respectively measured by Rhodamine123 and Fluo-3 AM Ester. Western blotting detects the involvement of several apoptosis-related proteins. Finally, caspase-3 and Caspase-9 activity are respectively examined. Results:, AFP-TF can induce apoptosis in Bel7402 and HepG2 AFP-positive hepatocarcinoma cells, but not SK-Hep-1 and Changliver cells. AFP-mRNA level changes little in apoptotic Bel7402 cells; while AFP expression is downregulated and uniformly dispersed throughout the whole cell. Addition of exogenous AFP or overexpression of intracellular AFP can reduce such apoptotic effect. Besides, apoptotic Bel7402 cells show a disruption of ,,m, an immediate elevation of Ca2+ concentration, a prominently decreased ratio of bcl-2 to bax, a release of cytochrome c from mitochondria to cytosol, and ultimately an activation of caspase-9 and caspase-3. Conclusion:, AFP-TF induced Bel7402 cells apoptosis is mitochondrial-dependent and is mediated by the interaction of AFP-TF with intracellular AFP. [source]

    Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y

    S. A. Killough
    Killough SA, Lundy FT, Irwin CR. Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y. International Endodontic Journal, 43, 835,842, 2010. Abstract Aim, To investigate whether dental pulp fibroblasts express neuropeptide Y (NPY) and NPY-Y1 in vitro and to determine the effects of the cytokines including interlukin-1, (IL-1,), TGF- ,1, substance P and NPY on the expression of NPY Y1. Methodology, Three primary fibroblast cell strains were obtained from freshly extracted human third molar teeth. RT-PCR was utilized to detect expression of NPY and mRNA expression. Membrane protein samples were isolated, and protein expression was determined by Western blotting. Radioimmunoassay was used to quantify NPY expression in healthy (n = 35) and carious (n = 39) whole pulp samples, and the student's t -test was used to test for statistical significance. In addition, the 3-(4,5-Dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assay fibroblast cell growth. Results, mRNA transcripts were found in all three fibroblast cell populations with the cytokines having a stimulatory effect on its expression (P < 0.05). NPY mRNA was not detected in the cell strains. NPY-Y1 receptor protein expression was visualized by Western blotting, and there was no effect of IL-1, or TGF- ,1 on its expression. The mean concentration of NPY-Ir determined by radioimmunoassay in non-carious teeth was 19.40 ng g,1 (±17.03 SD) compared to 29.95 ng g,1 (±20.99 SD) in carious teeth (P < 0.05). Conclusion, Human dental pulp fibroblasts express, but do not synthesize, NPY, demonstrating that the fibroblast is a target cell for NPY. The effect of proinflammatory cytokines suggests that fibroblasts play a neuroimmunomodulatory role in the pulpal response to dental caries and injury. [source]

    The cytotoxic effects of resin-based sealers on dental pulp stem cells

    O. Trubiani
    Trubiani O, Caputi S, Di Iorio D, D'Amario M, Paludi M, Giancola R, Di Nardo Di Maio F, De Angelis F, D'Arcangelo C. The cytotoxic effects of resin-based sealers on dental pulp stem cells. International Endodontic Journal. Abstract Aim, To evaluate the effect of four current resin-based adhesives on expanded ex vivo human dental pulp mesenchymal stem cells (DP-MSCs). Methodology, Dental pulp mesenchymal stem cells were derived from dental pulps of ten donors. After in vitro isolation, dental pulp stem cells were analysed using flow cytometry. The immunophenotype of DP-MSCs disclosed the homogeneous expression of the mesenchymal-related antigens CD29, CD44, CD73, CD90, CD105, CD166. DP-MSCs were exposed to four different commercially available bonding systems (CMF Bond, Prime&Bond NT, Clearfil S3 Bond, XP Bond), and after 24, 48 and 72 h of incubation the morphological features and the cell growth were analysed. Moreover, the cell viability was evaluated at the same times by MTT assay. Data were statistically analysed using a two-way anova and Holm,Sidak method (, set at 0.05). Results, Significant differences were observed between the four groups when comparing DP-MSCs appearance. DP-MSCs survived and proliferated without inhibition in the presence of CMF Bond adhesive. On the contrary, microscopic evaluation of the other three groups revealed extensive cytotoxic effects from the dentine bonding agents. The MTT assay revealed no statistically significant differences in cell viability after 72 h between the control group and CMF Bond group. All the other experimental groups had statistically lower optical density values. Conclusions, CMF Bond adhesive allowed human dental pulp stem cells to survive and proliferate. All of the other dentine bonding agents had extensive cytotoxic effects. [source]

    The biocompatibility of modified experimental Portland cements with potential for use in dentistry

    J. Camilleri
    Abstract Aim, To evaluate the biocompatibility of a group of new potential dental materials and their eluants by assessing cell viability. Methodology, Calcium sulpho-aluminate cement (CSA), calcium fluoro-aluminate cement (CFA) and glass,ionomer cement (GIC; Ketac Molar), used as the control, were tested for biocompatibility. Using a direct test method cell viability was measured quantitatively using alamarBlueÔ dye, and an indirect test method where cells were grown on material elutions and cell viability was assessed using methyltetrazolium (MTT) assay as recommended by ISO 10 993-Part 5 for in vitro testing. Statistical analysis was performed by analysis of variance and Tukey multi-comparison test method. Results, Elution collected from the prototype cements and the GIC cured for 1 and 7 days allowed high cell activity after 24 h cell exposure, which reduced after 48 h when compared to the nontoxic glass,ionomer control, but increased significantly after 72 h cell contact. Elutions collected after 28 days revealed reduced cell activity at all cell exposure times. Cells placed in direct contact with the prototype materials showed reduced cell activity when compared with the control. Conclusions, Cell growth was poor when seeded in direct contact with the prototype cements. GIC encouraged cell growth after 1 day of contact. The eluted species for all the cements tested exhibited adequate cell viability in the early ages with reduced cell activity at 28 days. Changes in the production of calcium hydroxide as a by-product of cement hydration affect the material biocompatibility adversely. [source]

    Vorinostat increases carboplatin and paclitaxel activity in non-small cell lung cancer cells

    Taofeek K. Owonikoko
    Abstract We observed a 53% response rate in non-small cell lung cancer (NSCLC) patients treated with vorinostat plus paclitaxel/carboplatin in a Phase I trial. Studies were undertaken to investigate the mechanism (s) underlying this activity. Growth inhibition was assessed in NSCLC cells by MTT assay after 72 hr of continuous drug exposure. Vorinostat (1 ,M) inhibited growth by: 17% ± 7% in A549, 28% ± 6% in 128-88T, 39% ± 8% in Calu1 and 41% ± 7% in 201T cells. Vorinostat addition to carboplatin or paclitaxel led to significantly greater growth inhibition than chemotherapy alone in all 4 cell lines. Vorinostat (1 ,M) synergistically increased the growth inhibitory effects of carboplatin/paclitaxel in 128-88T cells. When colony formation was measured after drug withdrawal, vorinostat significantly increased the effects of carboplatin but not paclitaxel. The % colony formation was control 100%; 1 ,M vorinostat, 83% ± 10%; 5 ,M carboplatin, 41% ± 11%; carboplatin/vorinostat, 8% ± 4%; 2 nM paclitaxel, 53% ± 11%; paclitaxel/vorinostat, 46% ± 21%. In A549 and 128-88T, vorinostat potentiated carboplatin induction of gamma-H2AX (a DNA damage marker) and increased ,-tubulin acetylation (a marker for stabilized mictrotubules). In A549, combination of vorinostat with paclitaxel resulted in a synergistic increase in ,-tubulin acetylation, which reversed upon drug washout. We conclude that vorinostat interacts favorably with carboplatin and paclitaxel in NSCLC cells, which may explain the provocative response observed in our clinical trial. This likely involves a vorinostat-mediated irreversible increase in DNA damage in the case of carboplatin and a reversible increase in microtubule stability in the case of paclitaxel. [source]