Mice and primary neuronal cultures. Hq, Apaf1-/-, and Bax-/- mice were maintained and genotyped as described previously (Fortin et al., 2001; Klein et al., 2002). Cortical neurons and cerebellar granule neurons (CGNs) were cultured from cortices of E15.5 mice and cerebellums of postnatal day 7 mice, respectively, as described previously (Fortin et al., 2001). Recombinant adenoviral vectors carrying human AIF or LacZ expression cassettes were prepared and used at 50 multiplicity of infection as described previously (Cregan et al., 2002).. Camptothecin treatment and cell viability assays. Neurons were treated with 10 μm camptothecin with or without 10 μm Boc-aspartyl (Ome)-fluoromethylketone (BAF) (Enzyme System Products, Livermore, CA) after 2 d in vitro (DIV). Cell survival was measured by the following: live/dead staining (Molecular Probes, Eugene, OR), Hoechst staining, MTT assay (Cell Titer kit; Promega, Madison, WI), terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end ...

TY - JOUR. T1 - Molecular cloning of a human cDNA encoding a novel protein, DAD1, whose defect causes apoptotic cell death in hamster BHK21 cells. AU - Nakashima, Torahiko. AU - Sekiguchi, Takeshi. AU - Kuraoka, Akio. AU - Fukushima, Kohtarou. AU - Shibata, Yosaburo. AU - Komiyama, Sohtaro. AU - Nishimoto, Takeharu. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 1993/10. Y1 - 1993/10. N2 - The tsBN7 cell line, one of the mutant lines temperature sensitive for growth which have been isolated from the BHK21 cell line, was found to die by apoptosis following a shift to the nonpermissive temperature. The induced apoptosis was inhibited by a protein synthesis inhibitor, cycloheximide, but not by the bcl-2-encoded protein. By DNA-mediated gene transfer, we cloned a cDNA that complements the tsBN7 mutation. It encodes a novel hydrophobic protein, designated DAD1, which is well conserved (100% identical amino acids between humans and hamsters). By comparing the base sequences ...

Introduction Internucleosomal cleavage of DNA is a hallmark of apoptosis. DNA cleavage in apoptotic cells can be detected in situ in fixed cells or tissue sections using the terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) method. TUNEL is highly selective for the detection of apoptotic cells but not necrotic cells or cells with DNA strand breaks resulting from irradiation or drug treatment. In the TUNEL assay, TdT enzyme catalyzes the addition of labeled dUTP to the 3 ends of cleaved DNA fragments. Hapten-tagged dUTP (e.g. digoxigenin-dUTP or biotin-dUTP) can be detected using secondary reagents (e.g. anti-digoxigenin antibodies or streptavi-din) for fluorescence or colorimetric detection. Alternatively, fluorescent dye-conjugated dUTP can be used for direct detection of fragmented DNA by fluorescence microscopy or flow cytometry. The TUNEL Andy Fluor™ 594 Apoptosis Detection Kit contains dUTP conjugated to biotin and Streptavidin conjugated to bright and ...

The purpose of this study is to determine if a preventive treatment with curcumin can protect intestinal epithelial cells from inflammatory damage induced by IFNγ. To achieve this goal we have used a human intestinal epithelial cell line (HT29) treated with IFNγ to undergo apoptotic changes that can reproduce the damage of intestinal epithelia exposed to inflammatory cytokines. In this model, we measured the effect of curcumin (curcuminoid from Curcuma Longa) added as a pre-treatment at different time intervals before stimulation with IFNγ. Curcumin administration to HT29 culture before the inflammatory stimulus IFNγ reduced the cell apoptosis rate. This effect gradually declined with the reduction of the curcumin pre-incubation time. This anti-apoptotic action by curcumin pre-treatment was paralleled by a reduction of secreted IL7 in the HT29 culture media, while there was no relevant change in the other cytokine levels. Even though curcumin pre-administration did not impact the activation of the

TY - JOUR. T1 - Mifepristone regulates expression of apoptosis related genes Fas and FasL in mouse endometrium. AU - Gao, F.. AU - Xu, F. H.. AU - Zhou, X. C.. AU - Han, X. B.. AU - Liu, Y. X.. PY - 2001/7/3. Y1 - 2001/7/3. N2 - AIM: To investigate the anti-implantation mechanism of mifepriston. METHODS: In situ hybridization and immunohistochemistry were applied to determine mRNA and protein. RESULTS: After mifepriston injection, the number of implantation sites were obviously reduced, mifepriston could inhibit the embryo implantation in mouse. The expression of apoptosis related genes, Fas and FasL, in mouse endometrium was also decreased after mifepriston treatment. CONCLUSION: The expression of apoptosis related genes Fas and FasL is regulated by mifepriston and the inhibitory effect of mifepriston on the embryo implantation may be mediated by action on the Fas/FasL system.. AB - AIM: To investigate the anti-implantation mechanism of mifepriston. METHODS: In situ hybridization and ...

title: Galpha12 Protects Vascular Endothelial Cells from Serum Withdrawal-Induced Apoptosis through Regulation of miR-155, doi: 10.3349/ymj.2016.57.1.247, category: Article

High-density lipoproteins (HDL) levels have been shown to be inversely correlated with coronary heart disease, but the mechanisms of the direct protective effect of HDL on endothelial cells are not fully understood. The apoptosis of endothelial cells induced by cytokines and/or oxidized low-density …

Vascular Endothelial Growth Inhibitor (VEGI) is an endothelial cell autocrine factor and a member of the tumor necrosis family of ligands. VEGI is able to specifically inhibit endothelial cell growth and is an efficient inhibitor of angiogenesis. The molecular mechanisms of VEGI activity on endothelial cells remain undefined. Here we focused on two important steps in the signal transduction of VEGI. We first determined a role of NF-κB in VEGI-induced apoptosis. We found that inhibition of the NF-κB pathway resulted in an increased apoptotic potential of VEGI. We conclude that the NF-κB pathway plays a role in suppressing the apoptotic potential of VEGI. We next investigated the receptor responsible for VEGI-induced endothelial cell apoptosis. DR3 is a receptor for VEGI and thus we first focused on confirming if DR3 is the receptor responsible for VEGI-mediated endothelial cell death. We determined VEGI had diminished apoptotic activity in endothelial cells that are depleted of DR3 by siRNA. ...

HOTAIR was known to enhance radioresistance in several cancers. However, the function of HOTAIR on radioresistance involving the regulation of HIF-1α in cervical cancer has not been reported. BALB/c nude mice were injected subcutaneously with HeLa cells and irradiated by X-ray. The tumor volume was measured and the expression of HOTAIR in tumors was detected by quantitative real-time PCR. Western blot was performed to detect the protein level of HIF-1α. MTT (3-(4,5-Dimethylthiazol-2-yl) 22,5-diphenyltetrazolium bromide) assay and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to examine the cell viability and cell apoptosis of HeLa cells and C33A cells exposed to radiation. Radiotherapy inhibited the tumor growth in mice bearing HeLa cells. Radiotherapy reduced the expression of HOTAIR and HIF-1α in tumor tissues and HeLa cells or C33A cells. HOTAIR overexpression abrogated the effect of radiation on the cell viability and cell apoptosis of HeLa and C33A cells

Methods and Findings: Studies were performed using wild type (WT) and A2 knockout (A2-/-) mice exposed to Oxygen Induced Retinopathy (OIR). Neuronal injury and apoptosis were assessed using immunohistochemistry, TUNEL (terminal deoxynucleotidyl transferase dUTP nick end) labeling and Western blotting. Electroretinography (ERG) was used to assess retinal function. Neuro-glial injury in WT ROP mice was evident by TUNEL labeling, retinal thinning, decreases in number of rod bipolar cells and glial cell activation as compared with room air controls. Significant reduction in numbers of TUNEL positive cells, inhibition of retinal thinning, preservation of the rod bipolar cells and prevention of glial activation were observed in the A2-/- retinas. Retinal function was markedly impaired in the WT OIR mice as shown by decreases in amplitude of the b-wave of the ERG. This defect was significantly reduced in A2-/- mice. Levels of the pro-apoptotic proteins p53, cleaved caspase 9, cytochrome C and the ...

Pure ginsenoside standards (saponins Rh2, PD, and PT), along with an Rh2-enhanced North American ginseng (Panax quinquefolius) leaf extract (LFRh2), were tested for cytotoxic activity in cultured THP-1 leukemia cells. Thermal treatment of ginseng leaf resulted in production of both Rh2 and Rg3 content that was confirmed by liquid chromatography-mass spectrometry (LC-MS). Flow cytometry of cells stained with annexin V-fluorescein isothiocyanate and propidium iodide showed that the LFRh2 significantly (p ≤ 0.05) increased apoptosis (18% ± 0.4%) after 23 h at a concentration that inhibited cell viability by 50% (LC50 (72 h) = 52 μmg/mL. In comparison, a similar significant (p ≤ 0.05) increase in apoptotic cell numbers occurred at 41 h of exposure for pure ginsenoside standards, PD (LC50 (72 h) = 13 μg/mL), PT (LC50 (72 h) = 19 μg/mL), and Rh2 (LC 50 (72 h) = 15 μg/mL). Although no further increase in apoptosis was observed in THP-1 cells after exposure to increasing concentrations of LFRh2 ...

Acute ultraviolet (UV) B exposure causes photokeratitis and induces apoptosis in corneal cells. Geranylgeranylacetone (GGA) is an acyclic polyisoprenoid that induces expression of heat shock protein (HSP)70, a soluble intracellular chaperone protein expressed in various tissues, protecting cells against stress conditions. We examined whether induction of HSP70 has therapeutic effects on UV-photokeratitis in mice. C57 BL/6 mice were divided into four groups, GGA-treated (500 mg/kg/mouse) and UVB-exposed (400 mJ/cm2), GGA-untreated UVB-exposed (400 mJ/cm2), GGA-treated (500 mg/kg/mouse) but not exposed and naive controls. Eyeballs were collected 24 h after irradiation, and corneas were stained with hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). HSP70, reactive oxygen species (ROS) production, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and protein kinase B (Akt) expression were also evaluated. Irradiated corneal

TY - JOUR. T1 - FADD/MORT1 is a common mediator of CD95 (Fas/APO-1) and tumor necrosis factor receptor-induced apoptosis. AU - Chinnaiyan, Arul M.. AU - Tepper, Clifford G. AU - Seldin, Michael F. AU - ORourke, Karen. AU - Kischkel, Frank C.. AU - Hellbardt, Stefan. AU - Krammer, Peter H.. AU - Peter, Marcus E.. AU - Dixit, Vishva M.. PY - 1996/3/1. Y1 - 1996/3/1. N2 - CD95 (Fas/APO-1) and tumor necrosis factor receptor-1 (TNFR-1) are related molecules that signal apoptosis. Recently, a number of novel binding proteins have been proposed to mediate the signaling of these death receptors. Here we report that an N-terminal truncation of one of these candidate signal transducers, FADD/MORT1, abrogates CD95-induced apoptosis, ceramide generation, and activation of the cell death protease Yama/CPP32. In addition, this dominant-negative derivative of FADD (FADD-DN) blocked TNF- induced apoptosis while not affecting NF-κB activation. FADD-DN bound both receptors, and in the case of CD95, it disrupted ...

In this paper, the use of (123)I-Annexin V for the detection of farnesyltransferase inhibitor (FTI)-induced apoptosis in tumour-bearing athymic mice is described. In vitro binding assays on LoVo cells show time- and dosage-dependent (125)I-Annexin V binding upon treatment with Tipifarnib (Zarnestra,

Paclitaxel is used to treat breast cancers, but the mechanisms by which it induces apoptosis are poorly understood. Consequently, we have studied the role of the FoxO transcription factors in determining cellular response to paclitaxel. Western blotting revealed that in a panel of nine breast cancer cell lines expression of ... read more FoxO1a and FoxO3a correlated with the expression of the pro-apoptotic FoxO target Bim, which was associated with paclitaxel-induced apoptosis. In MCF-7 cells, which were paclitaxel-sensitive, the already high basal levels of FoxO3a and Bim protein increased dramatically after drug treatment, as did Bim mRNA, which correlated with apoptosis induction. This was not observed in MDA-231 cells, which expressed low levels of FoxOs and Bim. Gene reporter experiments demonstrated that in MCF-7 cells maximal induction of Bim promoter was dependent on a FoxO binding site, suggesting that FoxO3a is responsible for the transcriptional up-regulation of Bim. Gene silencing ...

Objectives : We investigated the effect of Haein-tang(Hairen-tang) on short-term memory and apoptosis in dentate gyrus of the gerbils with transient global ischemia. Methods : For the induction of cerebral ischemia model in mice, common carotid arteries of gerbils were occluded with aneurysm clips for 5 min. One day after operation, Haein-tang(Hairen-tang) was administrated orally injected once a day for 15 consecutive days. Gerbils were randomly divided into four group(n=10 in each group): sham-operation group, ischemia-induction group, ischemia-induction and 50 mg/kg Haein-tang(Hairen-tang)-treated group, ischemia-induction and 100 mg/kg Haein-tang(Hairen-tang)-treated group, and ischemia-induction and 200 mg/kg Haein-tang(Hairen-tang)-treated group. The effect of Haein-tang(Hairen-tang) on memory function was investigated by using step-down avoidance task. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) staining and immunohistochemistry ...

We used cytoplasmic extracts from chicken DU249 cells at various stages along the apoptotic pathway to analyse the events of apoptotic exe-cution. So-called S/M extracts from morphologically normal committed-stage cells induce apoptotic morphology and DNA cleavage in substrate nuclei. These apoptotic changes appear to require the function of multiple caspases (cysteine aspar-tases, a specialized class of proteases) acting in parallel. Extracts from execution-stage apoptotic cells induce apoptotic events in added nuclei in a caspase-independent manner. Biochemical frac-tionation of these extracts reveals that a column fraction enriched in endogenous active caspases is un-able to induce DNA fragmentation or chromatin condensation in substrate nuclei, whereas a caspase-depleted fraction induces both changes. Execution-stage extracts contain an ICAD/DFF45-inhibitable nuclease resembling CAD, plus another activity that is required for the apoptotic chromatin condensation. Committed-stage S/M ...

As a process, apoptosis integrates a multitude of pro- and anti-cell death signals to a single decision for life or death. This type of all-or-none response to quantitative inputs is common in biological phenomena such as cell proliferation and differentiation and is often associated with a network topology containing positive feedback loops (28). While the ability of caspases to self-cleave provides one level of such feedback activity (17), we find that this feedback extends to cross activation between both the extrinsic and intrinsic pathways, with specific activation of either extrinsic or intrinsic apoptosis leading to near simultaneous activation of both caspase-8 and -9 in all apoptotic cells tested in this experiment. This supports a model wherein, although initiation of caspases is segregated by distinct pathways, an apoptotic signal, once having reached a threshold level, leads to a simultaneous activation across both arms of the apoptotic network. Several signaling mechanisms ...

In previous study we showed that caspase-2 plays the role of an apical caspase in cell death induction by taxanes in breast cancer cells. This study deals with the role of other caspases. We tested breast cancer cell lines SK-BR-3 (functional caspase-3) and MCF-7 (nonfunctional caspase-3). Using western blot analysis we demonstrated the activation of initiator caspase-8 and -9 as well as executioner caspase-6 and -7 in both tested cell lines after application of taxanes (paclitaxel, SB-T-1216) at death-inducing concentrations. Caspase-3 activation was also found in SK-BR-3 cells. Employing specific siRNAs after taxane application, suppression of caspase-3 expression significantly increased the number of surviving SK-BR-3 cells. Inhibition of caspase-7 expression also increased the number of surviving SK-BR-3 and MCF-7 cells. On the other hand, suppression of caspase-8 and caspase-9 expression had no significant effect on cell survival. However, caspase-9 seemed to be involved in the activation of

TY - JOUR. T1 - Calcineurin Aβ Gene Targeting Predisposes the Myocardium to Acute Ischemia-Induced Apoptosis and Dysfunction. AU - Bueno, Orlando F.. AU - Lips, Daniel J.. AU - Kaiser, Robert A.. AU - Wilkins, Benjamin J.. AU - Dai, Yan Shan. AU - Glascock, Betty J.. AU - Klevitsky, Raisa. AU - Hewett, Timothy E.. AU - Kimball, Thomas R.. AU - Aronow, Bruce J.. AU - Doevendans, Pieter A.. AU - Molkentin, Jeffery D.. N1 - Copyright: Copyright 2008 Elsevier B.V., All rights reserved.. PY - 2004/1/9. Y1 - 2004/1/9. N2 - Cardiovascular disease is the leading cause of mortality and morbidity within the industrialized nations of the world, with coronary heart disease (CHD) accounting for as much as 66% of these deaths. Acute myocardial infarction is a typical sequelae associated with long-standing coronary heart disease resulting in large scale loss of ventricular myocardium through both apoptotic and necrotic cell death. In this study, we investigated the role that the calcium calmodulin-activated ...

TY - JOUR. T1 - Nitrite generation and antioxidant effects during neutrophil apoptosis. AU - Misso, Neil L.A.. AU - Peacock, Craig D.. AU - Neil Watkins, D.. AU - Thompson, Philip J.. PY - 2000/3/15. Y1 - 2000/3/15. N2 - Neutrophil apoptosis is important for the resolution of airway inflammation in a number of lung diseases. Inflammatory mediators, endogenous reactive oxygen and nitrogen species, and intracellular and extracellular antioxidants may all influence neutrophil apoptosis. This study investigated the involvement of these factors during apoptosis of neutrophils cultured in vitro. Neutrophils undergoing spontaneous apoptosis in culture as assessed by annexin V binding generated significant amounts of nitrite. Incubation with agonistic anti-Fas monoclonal antibody or tumor necrosis factor-α (TNF-α) enhanced neutrophil apoptosis at 6 h, although it decreased nitrite accumulation. Although granulocyte-macrophage colony-stimulating factor significantly reduced neutrophil apoptosis, this ...

Caspase 3 is an essential death factor for the Fas-mediated cell death, and its inactivation in cells is initiated by an interaction with p21 on mitochondria or with IAP family member ILP. Survivin is also a member of IAP family and is specifically expressed during embryogenesis and in tumor cells and suppresses cell death signaling. In our current study, we demonstrated that Survivin translocation into the nucleus is dependent on Fas stimulation and cell proliferation. Survivin also interacts with the cell cycle regulator Cdk4, leading to Cdk2/Cyclin E activation and Rb phosphorylation. As a result of Survivin/Cdk4 complex formation, p21 is released from its complex with Cdk4 and interacts with mitochondrial procaspase 3 to suppress Fas-mediated cell death. Here, we propose that Survivin supports procaspase 3/p21 complex formation as a result of interaction with Cdk4 resulting in suppression of cell death signaling.

TY - JOUR. T1 - Pretreatment with low nitric oxide protects osteoblasts from high nitric oxide-induced apoptotic insults through regulation of c-Jun N-terminal kinase/c-Jun-mediated Bcl-2 gene expression and protein translocation. AU - Tai, Yu-Ting. AU - Cherng, Yih-Giun. AU - Chang, Chia Chen. AU - Hwang, Yi Ping. AU - Chen, Jui Tai. AU - Chen, Ruei-Ming. PY - 2007/5. Y1 - 2007/5. N2 - Nitric oxide (NO) can regulate osteoblast activity. In this study, we evaluated the effects of pretreatment with a low concentration of NO on osteoblast injuries induced by a high level of NO and its possible molecular mechanisms. Exposure of osteoblasts to 0.3 mM sodium nitroprusside (SNP), an NO donor, slightly increased cellular NO levels without affecting cell viability. SNP at 2 mM greatly increased the levels of cellular NO and reactive oxygen species, and induced osteoblast death. Thus, osteoblasts were treated with 0.3 and 2 mM SNP as the sources of low and high NO, respectively. Exposure of osteoblasts ...

Because defects in apoptosis programs, for example, high expression of antiapoptotic molecules, can cause resistance to treatment regimens including radiotherapy (8), current attempts to improve the outcome of glioblastoma patients depend on strategies to increase apoptosis sensitivity. In this study, we identify a new proapoptotic role of NF-κB in γ-irradiation-mediated apoptosis of glioblastoma cells by showing, for the first time, that NF-κB is critically required for Smac mimetic-triggered radiosensitization. This conclusion is supported by several independent pieces of evidence. First, BV6 and γ-irradiation cooperate to trigger apoptosis in glioblastoma cells. This interaction is highly synergistic in A172 glioblastoma cells (CI , 0.3). Second, BV6 stimulates NF-κB activation, which is required for the potentiation of γ-irradiation-induced apoptosis because genetic inhibition of NF-κB by overexpression of the dominant-negative superrepressor IκBα-SR significantly reduces BV6- and ...

TY - JOUR. T1 - Nitric oxide-GAPDH-Siah. T2 - a novel cell death cascade.. AU - Hara, Makoto R.. AU - Snyder, Solomon H. PY - 2006/7. Y1 - 2006/7. N2 - 1. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an extremely abundant glycolytic enzyme, and exemplifies the class of proteins with multiple, seemingly unrelated functions. Recent studies indicate that it is a major intracellular messenger mediating apoptotic cell death. This paper reviews the GAPDH cell death cascade and discusses its clinical relevance. 2. A wide range of apoptotic stimuli activate NO formation, which S-nitrosylates GAPDH. The S-nitrosylation abolishes catalytic activity and confers upon GAPDH the ability to bind to Siah, an E3-ubiquitin-ligase, which translocates GAPDH to the nucleus. In the nucleus, GAPDH stabilizes the rapidly turning over Siah, enabling it to degrade selected target proteins and affect apoptosis. 3. The cytotoxicity of mutant Huntingtin (mHtt) requires nuclear translocation which appears to be ...

3H]Thymidine Incorporation Studies. Thymidine incorporation studies were performed with cells grown in 35-mm culture dishes. The cells were labeled by the addition of methyl-[3H]thymidine (1 μCi/ml) to the medium, and the reactions were stopped after 1 h. The medium was drawn off, and the cells were rinsed twice with ice-cold phosphate-buffered saline (PBS). The cultures were then fixed with ice-cold 5% trichloracetic acid overnight at 4°C, after which the cells were extracted as previously described (Guo and Reiners, 2000). A second replicate set of nonfixed dishes was treated with 0.25% trypsin-EDTA to estimate cell numbers. [3H]Thymidine was detected by scintillation counting and expressed as disintegration per minute per 103 cells.. Assessment of Apoptosis. The effects of HET0016 to induced apoptosis in 9L cells were examined by fluorescence-activated cell sorting analysis after the cells were labeled with an annexin V-fluorescein isothiocyanate (FITC) antibody (Sigma Chemical, St. Louis, ...

Previous studies suggested that dithio-carbamates are potent apoptosis and anti-apoptosis inducing agents in various cancer cells. Here, the anti-proliferative and apoptosis inducing effects of a new derivative (2-NDC) from the dithio-carbamate family was examined in human leukemia K562 cells. We use thiazolyl blue tetrazolium bromide (MTT) to measure viability and cell growth inhibition. The 2-NDC showed effects on viability in a dose and time-dependent manner, inhibiting proliferation at concentrations of 10-30 M after 24-48 hours of treatment and increasing values after 72 hours at 40-120 M. The cytotoxic effect of the compound was calculated with an IC50 of 30 M after 24-hour. Apoptosis induction was confirmed by acridine orange-ethidium bromide (AO/EtBr) staining, DNA fragmentation assay, flow cytometric assessment and also caspase-3 activation assay. Furthermore, enzymes level such as superoxide dismutase (SOD) and catalase (CAT) involved in oxidative stress were evaluated. The results of this

Our results uncovered an unexpected role for MPO to influence the fate of neutrophils and consequently the duration of inflammation. By suppressing the constitutive cell death program, MPO prolonged the life span of neutrophils, thereby delaying the resolution of inflammation. These actions were specific for MPO, because other azurophilic granule constituents lactoferrin and elastase failed to affect neutrophil apoptosis.. Consistent with the commitment of neutrophils to apoptosis, MPO at clinically relevant concentrations delayed, rather than blocked apoptosis, resulting in prolonged survival of human neutrophils in vitro. We confirmed that increasing plasma concentrations of MPO in rats to levels comparable to those detected in patients with inflammatory vascular diseases20,21 was sufficient to retard the apoptotic machinery in neutrophils as assayed ex vivo. Furthermore, MPO also suppressed apoptosis in neutrophils that had emigrated into the airways and delayed resolution of inflammation in ...

TY - JOUR. T1 - The N-terminal helix of Bcl-xL targets mitochondria. AU - McNally, Melanie A.. AU - Soane, Lucian. AU - Roelofs, Brian A.. AU - Hartman, Adam L.. AU - Hardwick, J Marie. PY - 2013/3. Y1 - 2013/3. N2 - Anti- and pro-apoptotic Bcl-2 family members regulate the mitochondrial phase of apoptotic cell death. The mitochondrial targeting mechanisms of Bcl-2 family proteins are tightly regulated. Known outer mitochondrial membrane targeting sequences include the C-terminal tail and central helical hairpin. Bcl-xL also localizes to the inner mitochondrial membrane, but these targeting sequences are unknown. Here we investigate the possibility that the N-terminus of Bcl-xL also contains mitochondrial targeting information. Amino acid residues 1-28 of Bcl-xL fused to EGFP are sufficient to target mitochondria. Although positive charges and helical propensity are required for targeting, similar to import sequences the N-terminus is not sufficient for efficient mitochondrial import.. AB - ...

The effects of the acute phase proteins AGP and AAT on early (2-hour) and late (24-hour) apoptosis and inflammation after renal I/R were investigated. Both AGP and AAT administered at reperfusion decrease early as well as late apoptosis, as reflected by renal internucleosomal DNA cleavage, TUNEL histology, and caspase-like activities. In line, we previously demonstrated that abrogating acute early apoptosis with selective antiapoptotic agents prevents subsequent inflammation as well as secondary apoptosis caused by inflammation, whereas antiapoptotic treatment that is initiated after the onset of apoptosis does not reduce I/R-induced inflammation.1 In contrast, in the present study, treatment after 2 hours of reperfusion inhibited inflammation, which is supported by reports of anti-inflammatory effects mediated by AGP and AAT.12 13 Early primary19 20 as well as late secondary2 21 apoptosis after I/R has been reported to be caused by various means. The present results, showing that AGP and (to a ...

TY - JOUR. T1 - Response of Apoptosis Related Proteins to Running in Fluoride-Exposed Mice. AU - Canning, M.. AU - Zohoori, Fatemeh. AU - Valentine, Ruth. AU - Khan, Z.. AU - Ahmed, A.. AU - Amaral, A.. AU - Azevedo, Liane. AU - Buzalaf, Maria. AU - Fabricio, M.. AU - Fernandes, M.. AU - Maguire, Anne. PY - 2017. Y1 - 2017. M3 - Meeting Abstract. VL - 96A. JO - Journal of Dental Research. JF - Journal of Dental Research. SN - 0022-0345. ER - ...

An in vitro ischemia model was established and the effect of the metabolic inhibitors cycloheximide (CHX) and actinomycin D (ActD) on apoptosis in astrocytes under ischemia studied. CHX decreased by 75% the number of cells dying after 6 hr of ischemia compared with control cultures. TdT-mediated dUTP nick end labelling (TUNEL) staining of comparable cultures was reduced by 40%. ActD decreased cell death by 60% compared with controls. The number of TUNEL-positive cells was reduced by 38%. The nuclear shrinkage in TUNEL-positive astrocytes in control cultures did not occur in ActD-treated astrocytes, indicating that nuclear shrinkage and DNA fragmentation during apoptosis are two unrelated processes. Expression of bcl-2 (alpha and beta), bax, and Ice in astrocytes under similar ischemic conditions, as measured by quantitative reverse transcription-polymerase chain reaction, indicated that ischemia down-regulated bcl-2 (alpha and beta) and bax. Ice was initially down-regulated from 0 to 4 hr, ...

AIMS--To compare in situ end-labelling (ISEL) of apoptosis in lung carcinoma with quantitative and semiquantitative light microscopic assessment and ultrastructural observations. METHODS--ISEL of apoptosis was evaluated in 42 lung carcinomas (24 squamous cell carcinomas, 12 adenocarcinomas and six small cell carcinomas). Results were correlated semiquantitatively with the extent of apoptosis in haematoxylin and eosin stained sections, with apoptotic indices and with ultrastructural observations (nine cases). RESULTS--In each tumour type the extent of apoptosis identified by ISEL correlated with that observed on light and electron microscopy. Tumour cells undergoing apoptosis showed either uniform nuclear staining with a surrounding halo or peripheral nuclear membrane staining. The latter pattern was more prominent in small cell carcinoma and correlated ultrastructurally with early apoptosis. A variable proportion of apoptotic cells and apoptotic bodies were unlabelled. Necrotic tumour cells ...

TY - JOUR. T1 - Increased cardiomyocyte apoptosis and changes in proapoptotic and antiapoptotic genes bax and bcl-2 during left ventricular adaptations to chronic pressure overload in the rat. AU - Condorelli, Gianluigi. AU - Morisco, Carmine. AU - Stassi, Giorgio. AU - Notte, Antonella. AU - Farina, Felicia. AU - Sgaramella, Giuseppe. AU - De Rienzo, Assunta. AU - Roncarati, Roberta. AU - Trimarco, Bruno. AU - Lembo, Giuseppe. PY - 1999/6/15. Y1 - 1999/6/15. N2 - Background - Left ventricular hypertrophy (LVH) represents both an adaptive response to increased cardiac work load and a precursor state of heart failure. Recent evidence linked cardiac myocyte death by apoptosis with LVH and heart failure. It remained unclear, however, whether apoptosis participated in the transition from LVH to left ventricular dysfunction (LVD). Methods and Results - Cardiac myocyte apoptotic events and changes in apoptosis-specific genes were studied in a rat model of chronic pressure overload induced by ...

Animals and plants diverged over one billion years ago and evolved unique mechanisms for many cellular processes, including cell death. One of the most well-studied cell death programmes in animals, apoptosis, involves gradual cell dismantling and engulfment of cellular fragments, apoptotic bodies, through phagocytosis. However, rigid cell walls prevent plant cell fragmentation and thus apoptosis is not applicable for executing cell death in plants. Furthermore, plants are devoid of the key components of apoptotic machinery, including phagocytosis as well as caspases and Bcl-2 family proteins. Nevertheless, the concept of plant

When hypertonicity is imposed with sufficient intensity and acuteness, cells die. Here we investigated the cellular pathways involved in death using a cell line derived from renal epithelium. We found that hypertonicity rapidly induced activation of an intrinsic cell death pathway- release of cytochrome c and activation of caspase-3 and caspase-9-and an extrinsic pathway-activation of caspase-8. Likewise, a lysosomal pathway of cell death characterized by partial lysosomal rupture and release of cathepsin B from lysosomes to the cytosol was also activated. Relationships among the pathways were examined using specific inhibitors. Caspase inhibitors did not affect cathepsin B release into the cytosol by hypertonicity. In addition, cathepsin B inhibitors and caspase inhibitors did not affect hyper-tonicity-induced cytochrome c release, suggesting that the three pathways were independently activated. Combined inhibition of caspases and cathepsin B conferred significantly more protection from ...

Differentiation refers to the ability of cancer cells to revert to their normal counterparts, and its induction represents an important noncytotoxic therapy for leukemia, and also breast, prostate, and other solid malignancies. Flavonoids are a group of differentiation-inducing chemicals with a potentially lower toxicology profile than retinoids. Flavonoid-rich polyphenol fractions from the pomegranate (Punica granatum) fruit exert anti-proliferative, anti-invasive, anti-eicosanoid, and pro-apoptotic actions in breast and prostate cancer cells and anti-angiogenic activities in vitro and in vivo.. The results highlight an important, previously unknown, mechanism of the cancer preventive and suppressive potential of pomegranate fermented juice and pericarp extracts.. Journal of Medicinal Food, Volume: 7 Issue 1: July 7, 2004 ...

TY - JOUR. T1 - Reduced wall compliance suppresses Akt-dependent apoptosis protection stimulated by pulse perfusion. AU - Li, Manxiang. AU - Chiou, Kuan Rau. AU - Bugayenko, Artem. AU - Irani, Kaikobad. AU - Kass, David A.. PY - 2005/9/16. Y1 - 2005/9/16. N2 - Reduced arterial compliance and increased pulse pressure are common and major risk factors for cardiovascular disease. Here, we reveal a novel mechanism whereby loss of wall distensibility blunts endothelial cell protection to oxidant stress-induced apoptosis. Bovine aortic endothelial cells cultured in compliant or stiff silastic tubes were pulse perfused by arterial pressure/flow waveforms generated by a servo-pump. Pulse perfusion induced time-dependent Akt activation peaking ,6-fold after 2 hours in compliant tubes and a similar time course but half the magnitude in stiff tubes. This was accompanied by quantitatively similar disparities in phosphoinositide-3 kinase activation and in Akt-stimulated suppressors of apoptosis: glycogen ...

Worldwide Market Reports added Latest Research Report titled Global Apoptosis Assays Market Is Expected To Reach Around USD 6.96 Billion By 2025″ to its Large Report database. The report, the global Apoptosis Assays Market, demonstrates an all-encompassing investigation of the global Apoptosis Assays Market that appraises the market magnitude and evaluates the market assessment above the predicted time. The ruling players of the Apoptosis Assays Market are visible inside the report in company with thorough details referring to their revenue break-up, synopsis of the business, latest developments, product segmentation, and so on. The operative market players in the global Apoptosis Assays Market are further discussed in the report.. This Research will help you grow your Business: [Download free Sample PDF of This Research Report]. Some of the Major Apoptosis Assays Market Players Are:. Merck KGaA, Thermo Fischer Scientific, Bio-Rad Laboratories, GE Healthcare, Geno Technology, BioTek ...

The basal machinery controlling PCD appears to have been substantially conserved throughout metazoan evolution. The mammalian bcl‐2 gene family encodes cell death regulator proteins that exhibit structural and functional homology with the product of the death‐suppressing ced‐9 gene of C.elegans. Likewise, the ced‐3 killer gene of C.elegans encodes a member of the conserved caspase family of cysteine proteases, members of which play critical proapoptotic roles in mammalian apoptosis.. However, despite this conservation of the basal machinery between the nematode and mammals, neither ced‐9/bcl‐2 nor caspase homologues has yet been identified in the fruit fly, D.melanogaster. Indeed, the proapoptotic machinery so far defined in Drosophila via genetic screening comprises three genes, rpr, hid and grim that share little if any homology with known components of the mammalian death machinery and none at all with the nematode. The only clear link thus far between the apoptotic machinery in ...

In this paper we used a multiparametric approach to analyze extensively the events occurring during apoptotic cell death of thymocytes, and furthermore, we asked whether alterations in mitochondrial structure and function are occurring in early stages of apoptosis. A multiparametric quantitative analysis was performed on normal or apoptotic thymocytes emerging from a few-hour culture performed in culture medium or in the presence of dexamethasone. Simultaneous detection of light scattering properties, integrity of plasma membrane (trypan blue exclusion), chromatin condensation (AO/EB staining of entire cells or PI staining of nuclei), and DNA fragmentation (in situ nick-translation in apoptotic cells) allowed a precise analysis of the preapoptotic and apoptotic stages. Moreover a thorough study of mitochondrial transmembrane potential (delta psi m) assessed following in a time course study the uptake by apoptotic cells of the cationic lipophilic dye DiOC6(3) or the J-aggregate-forming cation ...

Successful wound closure is mainly the result of two cellular processes: migration and proliferation. Apoptosis has also been suggested to play a role in the mechanisms of wound healing. The fast calcium wave (FCW), triggered immediately after a wound is produced, has been proposed to be involved in determining healing responses in epithelia. We have explored the effects of the reversible inhibition of FCW on the apoptotic and proliferative responses of healing bovine corneal endothelial (BCE) cells in culture. The most important findings of this study are that caspase-dependent apoptosis occurs during the healing process, that the amount of apoptosis has a linear dependence on the migrated distance, and that FCW inhibition greatly increases the apoptotic index. We have further been able to establish that FCW plays a role in the control of cell proliferation during BCE wound healing. These results indicate that one of the main roles of the wave is to inhibit an excessive apoptotic response of the

CCAR1; cell division cycle and apoptosis regulator 1; cell division cycle and apoptosis regulator protein 1; CARP 1; CARP1; FLJ10590; death inducer with SAP domain; cell cycle and apoptosis regulatory protein 1; MGC44628; RP11-437A18.1; novel protein similar to vertebrate cell division cycle and apoptosis regulator 1 (CCAR1 ...

Cell apoptosis assay PaTu8988 cell apoptosis was Inhibitors,Modulators,Libraries detected by the Annexin V Apoptosis Detection Kit in accordance on the producers protocol. Briefly, 1 million cells with indicated therapies were stained with FITC Annexin V and PI. Each early and late apoptotic cells had been sorted by fluorescence activated cell sorting. Cell morphologic examination A total of 4 104 PaTu8988 cells were seeded on glass cover slips during the six very well plate and taken care of using the indicated concentration of SAHA for 48 h. Cells were fixed and stained with Wright Giemsa stain. The slides had been photographed using oil microscopy. In vitro tube formation assay or vasculogenic mimicry assay The tumor cell formation of capillary structure in vitro was examined as we previously described.. Cellular immuno fluorescence staining PaTu8988 cells were seeded on glass cover slips in merely six nicely plates and taken care of with described dosage of SAHA for 48 h. Cells about the ...

The development and growth of aneurysms involve the complex events of arterial wall cells remodeling (38). Previous studies have demonstrated that apoptosis is involved in the pathogenesis of aneurysms (6,7). However, the molecular mechanisms underlying the stimulation of apoptosis in the mouse IA model remains unknown.. To investigate the apoptotic events of IA mice model, the present study investigated the mRNA expression of caspases associated with the mitochondrial apoptotic pathway, including caspase-3, −8 and −9. The activation of caspases serves an essential role during the process of apoptosis (39), which may be caused by an extrinsic or intrinsic pathway, which lead to a terminal common pathway (40). Caspase-8 activation is involved in the extrinsic pathway and caspase-9 is involved in the intrinsic pathway. In the mitochondrial pathway, the release of Cyt-c constitutes an apoptotic complex, which subsequently results in the activation of caspase-9. The activation of caspase-8 and ...

The nuoG gene of Mycobacterium tuberculosis (Mtb) has the ability to inhibit host cell apoptosis. This ability is a virulence factor and does not exist in facultative pathogenic and non-pathogenic mycobacterial species. NuoG is part of the NDH-1 complex, and this study addressed the potential link between the role of NuoG in apoptosis inhibition and the biochemical activity of the NDH-1 complex. Different mycobacterial species were tested for their NDH-1 activities. Among the bacteria tested were bacteria transformed with the Mtb nuoG plasmid, or with the almost entire NDH-1 coding region. Surprisingly, the levels of NDH-1 activity did not correlate with apoptosis levels, suggesting a potential independent, novel mechanism by which NuoG inhibits host cell apoptosis. ...

Abstract: PURPOSE: The clinical use of the zoledronic acid has been increasing recently, and especially for the treatment of bone metastases. The synergistic effects of zoledronic acid with other chemotherapeutic agents have been shown. However it is not known whether a similar synergistic apoptotic effects exist for a combination therapy of zoledronic acid with radiation. METHODS: The MCF-7 human breast cancer cell lines were treated with 10 micrometer, 50 micrometer or 100 micrometer of zoledronic acid, irradiated with 5 Gy, and they were also treated with a combination of both treatments. The results of the synergistic effect on apoptosis were identified by performing XTT assay, DNA fragmentation assay, FACS analysis and western blot analysis at 24, 48, 72 hours after treatment. RESULTS: Zoledronic acid afftects anti-proliferative and apoptotic effects on MCF-7 breast cancer cell line in a dose-dependent manner. The synergistic cytotoxic effect of zoledronic acid and radiation was noted. The ...

Apoptotic signaling is altered at many loci in cancer cells. Although many tumors develop resistance to cytochrome c-induced apoptosis, we have discovered that breast cancer cells exhibit a unique hypersensitivity to cytochrome c-induced apoptosis. Interestingly, this sensitivity is not due to changes in core apoptosomal proteins ( 44) but is due to a PHAPI-mediated posttranslational event that enhances the recruitment of caspase-9 to the Apaf-1 CARD. These findings have the potential to affect breast cancer chemotherapy through the development of apoptosome activators or cytochrome c mimetics as shown, in principle, by the fact that malignant mammary epithelial cells could be more easily killed by cytosolic cytochrome c than their normal counterparts.. PHAPI-mediated increase in caspase activation in breast cancer. Whereas many inhibitory signaling pathways converge on the apoptosome, there are very few physiologic/pathologic examples of enhanced apoptosome activation. Our data suggest that ...

In this study we investigated the presence of placental apoptosis in pregnancies with intrauterine meconium passage. Placental tissue samples at term were obtained from 15 normal and 15 pregnancies with intrauterine meconium passage. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method was applied to confirm apoptosis in all placental samples. The mean apoptotic ratio was significantly higher in placentas of pregnancies with intrauterine meconium passage when compared with normal ones (0.37 = 0.06% and 0.29 +/- 0.07%, respectively; p < 0.01). The mean apoptotic ratios in trophoblastic and stromal cells were significantly higher in placentas of pregnancies with intrauterine meconium passage than normal placentas (0.47 +/- 0.10% and 0.36 +/- 0.11% [p < 0.05] and 0.29 +/- 0.07% and 0.22 +/- 0.07% [p < 0.05], respectively). Increased placental apoptosis in pregnancies with intrauterine meconium passage could be a causative factor in the pathophysiology of ...

Background: Captopril is an angiotensin-converting enzyme inhibitor with sulphydryl groups in its chemical structure. It is commonly used as an antihypertensive drug. The occurrence of pemphigus vulgaris has repeatedly been reported in patients receiving captopril. The capacity of captopril and pemphigus serum to induce acantholysis, in vivo or in vitro, has been demonstrated experimentally. Objectives: To show that captopril and pemphigus serum, acting by a biochemical and immunological mechanism, respectively, trigger apoptosis. Methods: Human keratinocyte cells were treated with 15 mmol L)1 captopril or with pemphigus serum. DNA was extracted and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling method was used to detect apoptosis. Results: DNA fragmentation occurred after 72 h of treatment. Increased expression of p53, c-myc and inducible nitric oxide (NO) synthase (iNOS) mRNA were observed by polymerase chain reaction (PCR) in the treated cells ...

Protein and mRNA expression of Sirt1 is significantly reduced by I/R. Cardiac-specific Sirt1-/- mice exhibited a significant increase (44±5% versus 15±5%; P=0.01) in the size of myocardial infarction/area at risk. In transgenic mice with cardiac-specific overexpression of Sirt1, both myocardial infarction/area at risk (15±4% versus 36±8%; P=0.004) and terminal deoxynucleotidyl transferase dUTP nick end labeling-positive nuclei (4±3% versus 10±1%; P,0.003) were significantly reduced compared with nontransgenic mice. In Langendorff-perfused hearts, the functional recovery during reperfusion was significantly greater in transgenic mice with cardiac-specific overexpression of Sirt1 than in nontransgenic mice. Sirt1 positively regulates expression of prosurvival molecules, including manganese superoxide dismutase, thioredoxin-1, and Bcl-xL, whereas it negatively regulates the proapoptotic molecules Bax and cleaved caspase-3. The level of oxidative stress after I/R, as evaluated by ...

TY - JOUR. T1 - Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines. AU - Nakagawa, Yoshihito. AU - Akao, Yukihiro. AU - Morikawa, Hiroshi. AU - Hirata, Ichiro. AU - Katsu, Kenichi. AU - Naoe, Tomoki. AU - Ohishi, Nobuko. AU - Yagi, Kunio. N1 - Funding Information: The DLD-1 and COLO201 colon cancer cell lines used in this study were kindly supplied by the Health Science Research Resources Bank of the Japan Health Sciences Foundation. This work was supported in part by a Grant-in-Aid for Scientific Research by the Ministry of Education, Science, Sports and Culture of Japan.. PY - 2002/3/29. Y1 - 2002/3/29. N2 - Exposure of three colon cancer cell lines, SW480, DLD-1, and COLO201, to arsenic trioxide in the medium induced a marked concentration-dependent suppression of cell growth. The intracellular contents of reduced glutathione (GSH) in these cell lines tended to be inversely correlated with the sensitivity of the cells to arsenic trioxide. Among the ...

In contrast to apoptosis induced by some anticancer drugs as well as other stimuli, such as UV irradiation, anti-CD3 stimulation, and mitogen treatment (54, 55, 56, 57, 58, 59), Tet induced T cell apoptosis through a mechanism independent of Fas/FasL interaction. This conclusion was supported by several observations. First, the expression of Fas/FasL mRNA was not enhanced by Tet treatment (our unpublished observations). Second, Tet did not increase Fas/FasL expression on the T cell surface (Fig. 5⇑). Third, both Fas-sensitive and Fas-resistant T cells showed comparable susceptibility to Tet-induced apoptosis (Fig. 6⇑). Lastly, Tet-induced activation of caspase-3 appeared to have similar kinetics and intensity in Fas-sensitive and Fas-resistant T cells (Fig. 8⇑). This observation suggests that Tet can activate the T cell death machinery without the participation of cell surface Fas/FasL molecules. Treatment with Tet might be useful in autoimmune disorders caused by mutations in Fas or FasL ...

Cell Lines and Reagents. UM-22A, UM-22B, and 1483 are human HNSCC cell lines (Lin et al., 2007a) and were grown in Dulbeccos modified Eagles medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and antibiotics. ABT-737 and A-793844 were provided by Abbott Laboratories (Abbott Park, IL), dissolved in DMSO, and stored at -80°C as 10 mM stocks. Cisplatin was obtained from the University of Pittsburgh Cancer Institute Pharmacy, and etoposide was from Sigma (St. Louis, MO). Lipofectamine 2000 reagent was obtained from Invitrogen (Carlsbad, CA), and Annexin V-fluorescein isothiocyanate apoptosis detection kits were from BD Biosciences (San Jose, CA). Anti-Bcl-2 antibody was from Dako (Glostrup, Denmark), and antibodies against Bcl-XL, Bax, Noxa, and Mcl-1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against PARP and Bak were obtained from Cell Signaling Technology (Danvers, MA), and anti-caspase-3 was from Assay Designs (Ann Arbor, MI). ...

Multiple sclerosis is one of the autoimmune diseases in the central nervous system. Multiple sclerosis occurs through multiple mechanisms, and it is also mediated in part by an apoptotic mechanism. Swimming exercise has been recommended for the prevention and treatment of chronic diseases. In the present study, we investigated the effects of swimming exercise on short-term memory in relation with apoptotic neuronal cell death in the hippocampus following induction of multiple sclerosis. For this study, step-down avoidance task, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, immunohistochemistry for caspase-3 were performed. The animal model of multiple sclerosis was made by bilateral intracerebral ventricle injection of ethidium bromide. The rats in the swimming exercise groups were forced to swim for 30 min once daily for 14 consecutive days, starting 3 days after induction of multiple sclerosis. In the present results, short-term memory was deteriorated in ...

Molecular and intra-cellular mechanisms involved in the regulation of apoptosis processes in endometrial cells are poorly understood and documented. We have investigated the possibility that Akt survival pathway might be involved in the regulation of apoptosis in the uterus during the estrous cycle. Rats with regular estrous cycle (4 days) were killed at different days of estrous cycle (diestrus, proestrus, estrus and metestrus). Uteri were collected and fixed for immunohistochemical staining (IHC) and apoptotic cell death detection by [TdT]-mediated deoxyuridinetriphosphate nick end-labelling (TUNEL) or endometrial protein extracts collected for Western analysis. TUNEL analysis revealed that apoptosis was mainly found at estrus compared to other day of estrous cycle. TUNEL positive cells were apparent in luminal epithelial cells only. No apoptotic cells were observed at proestrus. In contrast, proliferation was maximal at proestrus as confirmed with the expression of CDC47/MCM7 (a cell proliferation

It has been known that apoptotic morphological changes are observed in cell death caused by ER stress (Imaizumi et al., 2001). Caspases are activated to transmit apoptotic signals transcending the difference in species (Alnemri et al., 1996). In rodents, caspase-12 mediates apoptosis specifically in response to ER stress (Nakagawa et al., 2000). Although human caspase-12 gene is transcribed into mRNA, mature caspase-12 protein would not be produced, because the gene is interrupted by frame shift and premature stop codon (Fischer et al., 2002). Furthermore, it contains amino acid substitution in the critical site, which leads to loss of function in several caspases (Fischer et al., 2002). Thus, human caspase-12 does not seem to function in ER stress-induced apoptosis, and some other caspases with similar structure might substitute functionally for caspase-12 in humans. The caspase-12 gene is located within a region where caspase-1/ICE subfamily genes cluster (caspases 1, 4, 5, 12 in human and ...

TY - JOUR. T1 - Hydrogen peroxide signaling is required for glucocorticoid-induced apoptosis in lymphoma cells. AU - Tome, Margaret E.. AU - Jaramillo, Melba C.. AU - Briehl, Margaret M.. PY - 2011/12/1. Y1 - 2011/12/1. N2 - Glucocorticoid-induced apoptosis is exploited clinically for the treatment of hematologic malignancies. Determining the required molecular events for glucocorticoid-induced apoptosis will identify resistance mechanisms and suggest strategies for overcoming resistance. In this study, we found that glucocorticoid treatment of WEHI7.2 murine thymic lymphoma cells increased the steady-state [H 2O 2] and oxidized the intracellular redox environment before cytochrome c release. Removal of glucocorticoids after the H 2O 2 increase resulted in a 30% clonogenicity; treatment with PEG-CAT increased clonogenicity to 65%. Human leukemia cell lines also showed increased H 2O 2 in response to glucocorticoids and attenuated apoptosis after PEG-CAT treatment. WEHI7.2 cells that overexpress ...

TY - JOUR. T1 - Gene therapy and neurotrophic factor treatment for amyotrophic lateral sclerosis. AU - Abe, Koji. AU - Manabe, Yasuhiro. AU - Murakami, Tetsuro. PY - 2001/12/1. Y1 - 2001/12/1. N2 - Although excitotoxic and oxidative stress play important roles in spinal neuron death, the exact mechanisms are not fully understood. We examined cell damage of primary culture of 11-day-old rat spinal cord by addition of glutamate, nitric oxide (NO) or peroxynitrite (PN) with detection of terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick end labeling (TUNEL). With addition of glutamate, NOC18 (a slow NO releaser) or PN, TUNEL positive nuclei were found in spinal large motor neurons from 24 h, and the positive cell proportion greatly increased at 48 h in contrast to the vehicle. The present results suggest that both excitotoxic and oxidative stress play important role in the apoptotic pathway in cultured rat spinal neurons. To examine a possible protective effect of exogenous ...

To reconstruct the mechanisms for the vasoobliteration that transforms diabetic retinopathy into an ischemic retinopathy, we compared the occurrence of cell death in situ in retinal microvessels of diabetic and nondiabetic individuals. Trypsin digests and sections prepared from the retinas of seven patients (age 67 +/- 7 yr) with .9 +/- 4 yr of diabetes and eight age- and sex-matched nondiabetic controls were studied with the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) reaction which detects preferentially apoptotic DNA fragmentation. The count of total TUNEL+ nuclei was significantly greater in the microvessels of diabetic (13 +/- 12 per one-sixth of retina) than control subjects (1.3 +/- 1.4, P = 0.0016), as were the counts of TUNEL+ pericytes and endothelial cells (P , 0.006). The neural retinas from both diabetic and nondiabetic subjects were uniformly TUNEL-. Retinal microvessels of rats with short duration of experimental diabetes or galactosemia and ...

Critically ill patients are at higher risk having acute lung injury (ALI) and more often in need of parenteral nutrition. We sought to study whether preconditioning with representative of lipid emulsions for one week could benefit rats from ALI. Using a lipopolysaccharide (LPS)-induced ALI rat model and techniques such as polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. PGE2 production in the serum was highest in the LPS group, followed with Intralipid group, and the PGE2 level of these two groups was significantly (P | 0.05) higher than the rest. Intralipid conditioning caused significantly less production of LTB4 than the LPS, Clinoleic, or Omegaven group. In contrast to Intralipid, rats pretreated with Clinoleic or Omegaven significantly decreased their production of inflammatory mediators (IL-1 β, IL-6 and TNF-α), had less apoptosis in the lung tissues, and Omegaven greatly improved liver

TY - JOUR. T1 - An inducible autocrine cascade regulates rat hepatocyte proliferation and apoptosis responses to tumor necrosis factor-α. AU - Cosgrove, Benjamin D.. AU - Cheng, Connie. AU - Pritchard, Justin R.. AU - Stolz, Donna B.. AU - Lauffenburger, Douglas A.. AU - Griffith, Linda G.. PY - 2008/7/1. Y1 - 2008/7/1. N2 - Tumor necrosis factor-α (TNF) is an inflammatory cytokine that induces context-dependent proliferation, survival, and apoptosis responses in hepatocytes. TNF stimulates and enhances growth factor-mediated hepatocyte proliferation and survival following partial hepatectomy, but also acts in concert with other inflammatory cytokines of the innate immune response during viral infection to induce apoptosis in hepatocytes. In other epithelial cell types, TNF has recently been shown to stimulate autocrine release of transforming growth factor-α (TGF-α) and interleukin-1 (IL-1) family ligands. Here, we examine the role of these autocrine ligands in modulating TNF-induced ...

Purpose: : In the intrinsic apoptotic pathway, Bcl-2 family member protein Bax triggers cytochrome C release from mitochondria. Thus, it leads to an activation of the downstream caspase cascade ultimately resulting in apoptosis. The aim of the study was to investigate kinetics and up-regulation of this central pro-apoptotic factor in human corneal endothelial cells. Methods: : Human corneal endothelial cells (HCECs, 100% confluence) were incubated with the intrinsic apoptotic inducer Etoposide (a topoisomerase II inhibitor) in increasing concentrations (1.25, 2.5, 5.0 µg/ml). Bax protein level was determined within twelve hours using cell-based ELISA. These results were confirmed by Western blot. The rate of apoptotic HCECs was measured 6h, 12h, 24h and 48h after incubation with Etoposide by flow cytometric assay using Annexin V and Propidium Iodide. Results: : We detected a positive correlation between increasing Bax protein level and apoptotic HCECs. Specifically, HCECs showed an early ...

Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 µg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. In the present findings we showed that low concentration of DC (|2.0 µg/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 µg/mL) significantly (p | 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Further, the FasL-induced apoptotic features

The Food and Drug Administration (FDA) issued a warning a few years ago that pregnant women taking the selective serotonin reuptake inhibitor (SSRI) antidepressant paroxetine risk giving birth to infants with major birth defects, including heart abnormalities Now comes word that the same drug (sold as Paxil, Paxil CR, Seroxat, Pexeva, and generic paroxetine hydrochloride) carries another danger that could keep babies from being born in the first place. A new study just published in the online edition of the journal Fertility and Sterility concludes as many as fifty percent of all men taking the antidepressant could have damaged sperm and compromised fertility.. New York Presbyterian Hospital and Weill Cornell Medical Center researchers followed 35 healthy male volunteers who took paroxetine for five weeks. Then sperm samples from the men were studied using an assay called terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to evaluate whether there were missing pieces of genetic ...

The Food and Drug Administration (FDA) issued a warning a few years ago that pregnant women taking the selective serotonin reuptake inhibitor (SSRI) antidepressant paroxetine risk giving birth to infants with major birth defects, including heart abnormalities Now comes word that the same drug (sold as Paxil, Paxil CR, Seroxat, Pexeva, and generic paroxetine hydrochloride) carries another danger that could keep babies from being born in the first place. A new study just published in the online edition of the journal Fertility and Sterility concludes as many as fifty percent of all men taking the antidepressant could have damaged sperm and compromised fertility.. New York Presbyterian Hospital and Weill Cornell Medical Center researchers followed 35 healthy male volunteers who took paroxetine for five weeks. Then sperm samples from the men were studied using an assay called terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to evaluate whether there were missing pieces of genetic ...

Intracisternal administration of p-n-Octyphenol into neonatal rats causes hyperactivity with the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling(TUNEL)-positive cells in the mesencephalon where immunoreactivity for tyrosine hydroxylase is reduced by the chemical ...

We have shown previously that cruciferous vegetable constituent benzyl isothiocyanate (BITC) suppresses viability of cultured MCF-7 and MDA-MB-231 human breast cancer cells and retards mammary cancer development in MMTV-neu mice by causing apoptosis, but the mechanism of cell death is not fully understood. We now show that whereas p53 is dispensable for BITC-induced cell death, proapoptotic response to this promising chemopreventive agent is mediated by suppression of X-linked inhibitor of apoptosis (XIAP) protein expression. The BITC treatment increased levels of total and Ser15-phosphorylated p53 protein in MCF-7 cells, but the proapoptotic response to this agent was maintained even after knockdown of the p53 protein level. Exposure of MCF-7 and MDA-MB-231 cells to BITC resulted in a marked decrease in protein level of XIAP as early as 8 hours after treatment. Ectopic expression of XIAP conferred statistically significant protection against BITC-mediated cytoplasmic histone-associated ...

This project is designed to investigate the possible role of apoptosis as a mode of cell death in irradiated and tamoxifen-treated breast cancer cells and to study the potential for using these manipulations to enhance cell killing and, thus, improve radiation therapy of breast cancer. To date, six human breast cancer cell lines and nine human non-breast cancer lines have been treated in vitro with ionizing radiation or tamoxifen, and the ability of the cells to undergo apoptosis has been evaluated using gel electrophoresis. None of the breast lines and only three of the other lines show radiation-induced apoptosis with this assay. Expression of p53 and bcl-2 has been investigated in most of the lines. Studies with four breast cancer lines have shown that high dose tamoxifen or estradiol slows growth or causes cell death, and in the two lines studied to date this treatment increases radiation sensitivity. These data suggest that breast cancer cells may contain strong anti-apoptotic mechanisms or have

The present study may have important pathological and therapeutic implications because overgrowth of VSMCs is a pivotal etiologic factor in the development of atherosclerosis and restenosis after angioplasty.26-28 To date, inhibiting VSMC proliferation is among the most effective strategies for preventing their overgrowth and controlling neointimal thickening.14 Previous studies have shown that targeting Ras with negative regulators or blocking the Ras downstream pathways is able to effectively attenuate restenosis from balloon catheterization.14,15,29-33 Our recent studies have demonstrated that rMfn-2 is a powerful endogenous Ras inhibitor and that somatic gene transfer of rMfn-2 profoundly inhibits rat VSMC proliferation and balloon injury-induced neointima thickening in vivo by inhibiting the Ras-Raf-MEK-ERK/MAPK signaling pathway.17. In addition to inhibition of cell proliferation, growing evidence has indicated that apoptosis also plays an essential role in the control of neointimal ...

TY - JOUR. T1 - Assessment of secondary necrosis of Jurkat cells using a new microscopic system and double staining method with annexin V and propidium iodide.. AU - Honda, O.. AU - Kuroda, Masahiro. AU - Joja, I.. AU - Asaumi, Jun-Ichi. AU - Takeda, Yoshihiro. AU - Akaki, S.. AU - Togami, I.. AU - Kanazawa, Susumu. AU - Kawasaki, S.. AU - Hiraki, Y.. PY - 2000/2. Y1 - 2000/2. N2 - Using a new system developed by us for acquiring microscopic images automatically, we compared the morphological changes that apoptotic cells undergo with changes in the staining pattern of annexin V-enhanced green fluorescent protein (AV-EGFP) and propidium iodide (PI) in individual cells. Jurkat cells were treated with 5 mM CaCl2 alone, anti-Fas antibody and heating at 42 degrees C for 30 min or 46 degrees C for 60 min, and then were incubated in medium with 5 mM CaCl2. Time-lapse DNA fragmentation analysis and morphological observation revealed that the anti-Fas antibody and heating at 42 degrees C for 30 min ...

Ezrin belongs to the ERM (ezrin-radixin-moesin) protein family and has been demonstrated to regulate early steps of Fas receptor signalling in lymphoid cells, but its contribution to TRAIL-induced cell death regulation in adherent cancer cells remains unknown. In this study we report that regulation of FasL and TRAIL-induced cell death by ezrin is cell type depen-dant. Ezrin is a positive regulator of apoptosis in T-lymphoma cell line Jurkat, but a negative regulator in colon cancer cells. Using ezrin phosphorylation or actin-binding mutants, we provide evidence that negative regulation of death receptor-induced apoptosis by ezrin occurs in a cytoskeleton-and DISC-independent manner, in colon cancer cells. Remarkably, inhibition of apoptosis induced by these ligands was found to be tightly associated with regulation of ezrin phosphorylation on serine 66, the tumor suppressor gene WWOX and activation of PKA. Deficiency in WWOX expression in the liver cancer SK-HEP1 or the pancreatic Mia PaCa-2 cell lines

Bis-(2,3-dibromo-4,5-dihydroxy-phenyl)-methane (BDDPM) is a bromophenol 1st isolated from 0. The migrated amount of HUVECs. The info proven in the graphs will be the mean SD beliefs of at least three specific tests. ** 0.01 control. 2.3. BDDPM Inhibits Vessel Sprouting in Vitro Following, we utilized spheroid capillary sprouting assay to review the result of BDDPM … Continue reading Bis-(2,3-dibromo-4,5-dihydroxy-phenyl)-methane (BDDPM) is a bromophenol 1st isolated from 0. The migrated. ...

X-linked inhibitor of apoptosis protein (XIAP), survivin, and BRUCE are members of the inhibitor-of-apoptosis protein (IAP) family known for their inhibitory effects on caspase activity and dysregulation of these molecules has widely been shown to cause embryonic defects and to promote tumorigenesis in human. Besides the anti-apoptotic functions, recent discoveries have revealed that XIAP, survivin, and BRUCE also exhibit regulatory functions for autophagy in cells. As the role of autophagy in human diseases has already been discussed extensively in different reviews; in this review, we will discuss the emerging autophagic role of XIAP, survivin, and BRUCE in cancer cells. We also provide an update on the anti-apoptotic functions and the roles in maintaining DNA integrity of these molecules. Second mitochondria-derived activator of caspases (Smac) is a pro-apoptotic protein and IAPs are the molecular targets of various Smac mimetics currently under clinical trials. Better understanding on the functions

TY - JOUR. T1 - Angiopoietin-1 protects endothelial cells from hypoxia-induced apoptosis via inhibition of phosphatase and tensin homologue deleted from chromosome ten. AU - Lee, Sae Won. AU - Youn, Seock Won. AU - Kim, Tae Youn. AU - Suh, Jung Won. AU - Koh, Gou Young. AU - Kwon, Yoo Wook. AU - Chae, In Ho. AU - Park, Young Bae. AU - Kim, Hyo Soo. PY - 2009/2. Y1 - 2009/2. N2 - Background and Objectives: Angiopoietin-1 (Ang1) is a regulator of blood vessel growth and maturation, and prevents radiation-induced or serum deprivation-induced apoptosis. Phosphatase and tensin homologue deleted from chromosome ten (PTEN), a well-known tumor suppressor, regulates cell cycle arrest and apoptosis. Hypoxia induces apoptosis by increasing the expression of PTEN. We hypothesized that Angl may regulate PTEN expression and, thus, reduce endothelial apoptosis under hypoxia in vitro and in vivo. Materials and Methods: In vitro, human umbilical vein endothelial cells (HUVECs) were treated with Ang1, and ...

The occurrence of in vivo apoptosis was investigated in lymph node sections obtained from HIV-infected persons at different stages of disease. The degree of apoptosis in lymph nodes from HIV-infected individuals was compared with that observed in lymph nodes obtained from HIV-negative individuals. Apoptosis was readily detected in lymph nodes obtained from both HIV-negative and HIV-positive persons; however, the degree of apoptosis in lymph nodes obtained from HIV-positive persons was three to four times higher than that observed in the lymph nodes obtained from HIV-negative persons. In contrast to HIV-negative lymph nodes in which apoptosis was confined largely to germinal centers, in HIV-positive lymph nodes all functional compartments of the lymph node (i.e., cortex, paracortex, and sinuses) were extensively involved by this phenomenon. Furthermore, a significant correlation was observed between intensity of apoptosis and degree of activation of the lymphoid tissue associated with HIV ...

TFAR19(TF-1 cell apoptosis related gene 19) is a novel apoptosis-related gene cloned from human leukemia cell line TF-1 cells undergoing apoptosis in 1999 (accession number AF014955 in GenBank). The human TFAR19 encodes a protein which shares significant homology to the corresponding proteins of species ranging from yeast to mice. TFAR 19 exhibits a ubiquitous expression pattern and its expression is upregulated in tumor cells undergoing apoptosis. Overexpression of TFAR 19 could enhance apoptosis of some tumor cells induced by growth factor withdrawal or serum deprivation. But the exact mechanism of TFAR 19 is unclear. Mitochondria not only provides energy for the cell, but also plays a critical rate on cell death or survival. The release of apoptosis promoting factor, such as cytochrome c from mitochondria, resulted by the damage of mitochondrial membrane integrity, is the key factor controlling apoptosis. The permeability transition pore (FIT) of mitochondria is a protein complex located ...

Apoptosis or programmed cell death plays a critical role in both inflammatory and immune responses. Recent evidence demonstrates that control of leukocyte apoptosis is one of the most striking immune system-related roles of melatonin. For this reason, this study evaluated the protective effects of melatonin on human leukocyte apoptosis induced by sustained cytosolic calcium increases. Such protective effects are likely mediated by melatonins free-radical scavenging actions. Treatments with the specific inhibitor of cytosolic calcium re-uptake, thapsigargin (TG), and/or the calcium-mobilizing agonist, N-formyl-methionyl-leucyl-phenylalanine (FMLP), induced intracellular reactive oxygen species (ROS) production, caspase activation as well as DNA fragmentation in human leukocytes. Also, TG- and/or FMLP-induced apoptosis was dependent on both cytosolic calcium increases and calcium uptake into mitochondria, because when cells were preincubated with the cytosolic calcium chelator, dimethyl BAPTA, and the

Apoptosis is programed cell death characterized by certain cellular changes and regulated by various gene products including Bcl-2 and caspase-1. The marijuana cannabinoid, Delta9tetrahydrocannabinol (THC), has been reported to suppress in culture the proliferation of splenocytes and increase the release of IL-1 from macrophages; however, the mechanisms of these effects remain unclear. Because cannabinoids have also been reported to induce apoptosis and because the release of IL-1 and suppression of lymphoproliferation are related to apoptosis, we tested for the induction of apoptosis by THC in murine immune cell cultures. Splenocytes cultured with Con A for up to 24 hr showed evidence of DNA fragmentation determined by gel electrophoresis, terminal deoxynucleotide transferase-mediated dUTP-fluorescein nick end labeling and 3H-thymidine labeling and THC (15-30 microM) treatment increased fragmentation under these conditions. Resident peritoneal macrophages cultured with lipopolysaccharides ...

Breast cancer is the most common cancer types among women. Recent researches have focused on determining the efficiency of alternative molecules and miRNAs in breast cancer treatment. The aim of this study was to determine the effect of usnic acid response-miR-185-5p on proliferation in the breast cancer cell and to determine its relationship with apoptosis pathway. The cell proliferation and cell apoptosis rate were significantly increased following the ectopic expression of miR-185-5p in BT-474 cells. Furthermore, the results of cell cycle assay performed by flow cytometry revealed that the transfection with miR-185-5p induced G1/S phase arrest. The apoptosis-related genes expression analysis was performed by qRT-PCR and the direct target of miR-185-5p in BT-474 cells was identified by western blot and luciferase reporter assay. Our data showed that miR-185-5p can cause significant changes in apoptosis-related genes expression levels, suggesting that cell proliferation was suppressed by miR-185-5p via

Full Text - The rapid and efficient clearance of apoptotic germ cells (GCs) by Sertoli cells (SCs) is important for spermatogenesis. High mitochondrial activity in phagocytes is critical for continued clearance of apoptotic cells. However, the underlying molecular mechanism is poorly understood. Glycogen synthase kinase-3α (GSK3α) is a protein kinase that participates in the regulation of mitochondrial activity. Immunohistochemistry evidenced the predominant presence of the Ser21 phosphorylation GSK3α (inactivation) signal in SCs. Heat shock-induced apoptosis of GCs and dephosphorylation of GSK3α in SCs is a perfect model to investigate the role of GSK3α in phagocytic action. The number of apoptotic GCs was significantly lower in GSK3α inhibitor pre-treated mice with HS compared to normal control. In vitro phagocytosis assays shown that the phagocytic activity in GSK3α activated SCs was downregulated, while GSK3α inhibitor supplementation restored this process. Moreover, GSK3α activation

Programmed cell death (PCD) and cell type involving dying cells in the developing mouse and human fetal kidneys were investigated using the modified TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method in combination with immunohistochemistry. We identified many TUNEL-positive apoptotic cells mainly in the nephrogenic region in the developing mouse and human fetal kidneys. Tissue-fixed macrophages were widely distributed throughout the developing mouse and human fetal kidneys, including the nephrogenic zone. Most of the apoptotic cells in the nephrogenic zone were phagocytosed by tissue-fixed macrophages, suggesting that tissue-fixed macrophages play an important role in the process of PCD in the developing mouse kidney and human fetal kidney.

ethyl ahcohol, respectively. After 2 hours, their blood were sampled with their consents. Fraction of lymphocytes was isolated by density gradient method with Histopaque-1077 (Sigma) and irradiated with dose from 0.5 to 5 Gy. After 60 hour incubation, the cells were harvested and analysed by flow cytometry. Cell viability was decreased by dose dependent manner. Cell viability of beer group was reduced about 15% compared with control group. Apoptosis in soju group was reduced about 20% compared with control group. Apoptosis of beer and control groups are similar. Necrosis of soju group significantly increased about 35% compared with control group. Early apoptosis of beer group was increased compared with control group. Early apoptosis of soju group was decreased about 25% compared with control group. Late apoptosis of beer and control group was increased by dose dependent manner. Late apoptosis of soju group was increased about 20-30% compared with control group. Late apoptosis of soju was ...

Background:Apoptosis or programmed cell death has been shown to play an important role in the progression from polyps to carcinomas. Fas/APO-1 is a cell surface protein that can induce apoptosis in...

We have found that GLNPs treatment selectively induced apoptosis in OSCC compared to the normal fibroblast cells. This selective effect in OSCC is achieved through enhanced reactive oxygen species (ROS) generation followed by Bax mitochondrial translocation and caspase 3 activation. Further, GLNPs induced withdrawal of epigenetic transcription repression through concurrent downregulation of the polycomb group proteins (PcG) Bmi 1 and EZH2 along with their successive targets, UbH2AK119 and H3K27me3, which have immense therapeutic implications in the treatment of OSCC. Last, we have established that GLNPs regulate EZH2expression through proteasomal mediated degradation and 3PK inhibition; 3PK protein was found physically linked with EZH2 protein and its promoter region (-1107 to -1002). This event indicates that 3PK might play some crucial role in EZH2 expression and epigenetic control of OSCC. Moreover, the formulation showed improved biodistribution, aqueous dispersibility and enhanced ...

TY - JOUR. T1 - Melatonin induces apoptosis through a caspase-dependent but reactive oxygen species-independent mechanism in human leukemia Molt-3 cells. AU - Perdomo, Juan. AU - Cabrera, Javier. AU - Estévez, Francisco. AU - Loro, Juan. AU - Reiter, Russel J.. AU - Quintana, José. PY - 2013/9/1. Y1 - 2013/9/1. N2 - Melatonin is a naturally occurring indoleamine synthesized in the pineal gland that exhibits an extensive repertoire of biological activities. An increasing number of studies indicate that melatonin protects normal cells, while it reducing cancer cell proliferation. In this study, we investigated the effect of melatonin on the growth of the human leukemia cells and found that it efficiently reduced the number of cells in a concentration- and timedependent manner. Thus, incubation with the indoleamine increased the percentage of cells with a hypodiploid DNA content, augmented the number of annexin V-positive cells, and also provoked ultrastructural changes that are features of ...

Methods Male wild type mice were randomly divided into control and PD150606 groups. Mice were subjected to myocardial ischemia by occlusion of the left anterior descending coronary artery for 45 min and reperfusion for 3 h (I/R). Terminal deoxynucleotidyl transferase d-UTP nick end labeling (TUNEL) staining was performed using an In Situ Cell Death Detection kit on paraffin heart sections (5 mm). Hoechst 33342 was used as a counter-stain. Adult mouse cardiomyocytes were isolated and cultured, then subjected to ischemia for 1 h, and reperfusion for 3 h. The survival of cardiomyocytes, activity of calpain and caspase-3, cytoplasmic DNA fragments and cytochrome c concentrations were determined. Results Compared to control, the numbers of TUNEL-positive nuclei were significantly increased in the peri-infarct area in I/R mice (p,0.05). Compared to normal cultured cardiomyocytes, the survival of the cells significantly decreased, however the activation of calpain and caspase-3, and cytoplasmic DNA ...

TY - JOUR. T1 - Inhibition of human neuroblastoma cell growth by CAY10404, a highly selective Cox-2 inhibitor. AU - Parashar, Bhupesh. AU - Shankar, Sai Latha. AU - OGuin, Kathleen. AU - Butler, James. AU - Vikram, Bhadrasain. AU - Shafit-Zagardo, Bridget. PY - 2005/1/1. Y1 - 2005/1/1. N2 - Neuroblastomas constitute about 10% of childhood cancers and are responsible for 15% of pediatric cancer mortality. We evaluated the efficacy and the mechanism of cell death induced by CAY10404, a selective cyclooxygenase-2 (Cox-2) inhibitor in four human neuroblastoma cell lines (SH-EP, SH-SY5Y, SK-N-MC and MSN). Treatment with CAY10404 in the range of 15-115 μM revealed a dose-dependent decrease in cell number and an average IC50(inhibitory concentration 50%) of 60 μM. About 20-30% of the cells were terminal deoxynucleotidyltransferase-mediated UTP nick-end-labeling (TUNEL) positive 48 h after treatment. Western blot analysis of CAY10404-treated cells showed poly(ADP-ribose) polymerase (PARP) cleavage ...

Pancreas ductal adenocarcinoma (PDAC) has the most dismal prognosis among all human cancers since it is highly resistant to chemotherapy, radiotherapy and immunotherapy. The anticipated consequence of all therapies is induction of tumor apoptosis. The highly resistance nature of PDACs to all therapies suggests that the intrinsic tumor cell factors, likely the deregulated apoptosis pathway, are key mechanisms underlying PDAC non-response to these therapies, rather than the therapeutic agents themselves. The aim of this study is to test the hypothesis that epigenetic dysregulation of apoptosis mediators underlies PDAC resistance to gemcitabine, the standard chemotherapy for human PDAC. PDAC cells were analyzed for apoptosis sensitivity in the presence of a selective epigenetic inhibitor. The epigenetic regulation of apoptosis regulators was determined by Western Blotting and quantitative PCR. The specific epigenetic modification of apoptosis regulator promoter chromatin was determined by chromatin

Apoptosis is a controlled form of cellular demolition, catalyzed by a family of cysteine proteases called caspases. In response to diverse proapoptotic stimuli, caspase-9 is recruited and activated within an oligomeric complex called the apoptosome. The apoptosome drives autocatalytic processing of caspase-9, triggering a proteolytic caspase cascade that results in the biochemical and morphological changes characteristic of cell death. It is unclear why caspase-9 undergoes autocatalytic processing following apoptosome recruitment, because interdomain processing is dispensable for caspase-9 activity. A study has shed light on this issue by demonstrating that caspase-9 processing within the apoptosome promotes its displacement from the complex, leading to inactivation of this protease. Thus, autoprocessing of caspase-9 within the apoptosome serves as a molecular timer that limits the proteolytic activity of this complex through displacement of bound caspase-9 molecules. This timer mechanism may ...

TY - JOUR. T1 - RACK1 downregulates levels of the pro-apoptotic protein Fem1b in apoptosis-resistant colon cancer cells. AU - Subauste, M. Cecilia. AU - Ventura-Holman, T.. AU - Du, L.. AU - Subauste, Jose S.. AU - Chan, Shing Leng. AU - Yu, Victor C.. AU - Maher, Joseph F.. PY - 2009/12/1. Y1 - 2009/12/1. N2 - Evasion of apoptosis plays an important role in colon cancer progression. Following loss of the Apc tumor suppressor gene in mice, the gene encoding Fem1b is upregulated early in neoplastic intestinal epithelium. Fem1b is a pro-apoptotic protein that interacts with Fas, TNFR1 and apaf-1, and increased expression of Fem1b induces apoptosis of cancer cells. Fem1b is a homolog of FeM-1, a protein in Caenorhabditis elegans that is negatively regulated by ubiquitination and proteasomal degradation. To study Fem1b regulation in colon cancer progression, we used apoptotis-sensitive SW480 cells, derived from a primary colon cancer, and their isogenic, apoptosis-resistant counterparts sW620 cells, ...

Nitric oxide-releasing non steroidal anti-inflammatory drugs (NO-NSAIDs) are a promising class of compounds that cause cell cycle perturbations and induce apoptosis in cell lines from different tumors. We investigated the activity of a recently developed NO-NSAID (NCX 4040) in bladder cancer cell lines (HT1376 and MCR). Cells were treated with different drug concentrations for different exposure times. Cytostatic and cytocidal activity was tested by SRB assay and apoptosis was evaluated by TUNEL analysis, ANNEXIN V assay and fluorescence microscopy. To further investigate the cell death-inducing mechanisms of NCX 4040, we analyzed gp-170, caspase expression and mitochondrial membrane potential (Delta Psi) depolarization. NCX 4040 showed a striking cytocidal activity in both cell lines, reaching LC(50) at a 10-mu M and 50-mu M concentrations in HT1376 and in MCR cells, respectively, after an exposure of only 6 h followed by an 18-h washout. Apoptosis was triggered in up to 90% of cells and was ...

We have recently shown that the co-expression and interaction of the receptor for advanced glycation end products (RAGE) and its ligand S100B post myocardial infarction play a role in myocyte apoptosis. In other cell types, the S100B/RAGE interaction triggers signaling pathways including NF-kB activation and translocation, MAP kinases, and p53. To determine the signaling pathways modulated by the S100B/RAGE interaction contributing to myocyte apoptosis, rat neonatal cardiac myocyte cultures transfected with a full-length cDNA of the RAGE gene, a dominant-negative cytoplasmic deletion mutant of RAGE, or vector control were treated with nanomolar (1-1000) concentrations of recombinant bovine brain S100B for 48 hrs. 1 μM S100B induced vector alone-transfected myocyte apoptosis, as evidenced by increased terminal deoxynucleotidyltransferase-mediated UTP end labeling (TUNEL) [12.4±0.7% vs. 5.6±0.3% (untreated) of total cells p,0.05], DNA fragmentation, cytochrome C release from the mytochondria to ...