Co-localization of active caspase-3 and DNA fragmentation (TUNEL) in normal and hyperthermia-induced abnormal mouse development.
AuthorsMirkes PE, Little SA, Umpierre CC
JournalTeratology
PubMed ID11283970
'BACKGROUND: Previous work has shown that caspase-3 activation and DNA fragmentation, two hallmarks of apoptosis, are induced in day 9 mouse embryos exposed to hyperthermia (43 degrees C); however, the methods used to assess caspase-3 activation (Western blot) and DNA fragmentation (gel electrophoresis) did not allow these apoptotic events to ... More
Cytochrome c-mediated apoptosis in cells lacking mitochondrial DNA. Signaling pathway involving release and caspase 3 activation is conserved.
AuthorsJiang S, Cai J, Wallace DC, Jones DP
JournalJ Biol Chem
PubMed ID10514472
'Mitochondria serve as a pivotal component of the apoptotic cell death machinery. However, cells that lack mitochondrial DNA (rho(0) cells) retain apparently normal apoptotic signaling. In the present study, we examined mitochondrial mechanisms of apoptosis in rho(0) osteosarcoma cells treated with staurosporine. Immunohistochemistry revealed that rho(0) cells maintained a normal ... More
Assembly of the nuclear transcription and processing machinery: Cajal bodies (coiled bodies) and transcriptosomes.
AuthorsGall JG, Bellini M, Wu Z, Murphy C
JournalMol Biol Cell
PubMed ID10588665
'We have examined the distribution of RNA transcription and processing factors in the amphibian oocyte nucleus or germinal vesicle. RNA polymerase I (pol I), pol II, and pol III occur in the Cajal bodies (coiled bodies) along with various components required for transcription and processing of the three classes of ... More
RNA antisense abrogation of MAT1 induces G1 phase arrest and triggers apoptosis in aortic smooth muscle cells.
AuthorsWu L, Chen P, Hwang JJ, Barsky LW, Weinberg KI, Jong A, Starnes VA
JournalJ Biol Chem
PubMed ID10026172
'The human MAT1 gene (ménage à trois 1) is an assembly factor and a targeting subunit of cyclin-dependent kinase (CDK)-activating kinase. The novel mechanisms by which MAT1 forms an active CDK-activating kinase and determines substrate specificity of CDK7-cyclin H are involved in the cell cycle, DNA repair, and transcription. Hyperplasia ... More
Apoptosis of epithelial cells and macrophages due to infection with the obligate intracellular pathogen Chlamydia psittaci.
AuthorsOjcius DM, Souque P, Perfettini JL, Dautry-Varsat A
JournalJ Immunol
PubMed ID9780196
'We have characterized the cytotoxic activity of the obligate intracellular bacterium Chlamydia psittaci, which resides within a membrane-bound vacuole during the 2-day infection cycle. We have established that infected epithelial cells and macrophages die through apoptosis, which is measurable within 1 day of infection and requires productive infection by the ... More
Endogenous glucocorticoids induced by a chemical stressor (ethanol) cause apoptosis in the spleen in B6C3F1 female mice.
AuthorsCollier SD, Wu WJ, Pruett SB
JournalToxicol Appl Pharmacol
PubMed ID9465277
'Stress-induced increases in glucocorticoid levels can cause-apoptosis in immature thymocytes, but it is not known if glucocorticoids at these levels can also cause apoptosis in peripheral lymphocytes. In the present study, mice were exposed to ethanol (EtOH) in a model designed to represent binge drinking. This induces a substantial stress ... More
The CNC basic leucine zipper factor, Nrf1, is essential for cell survival in response to oxidative stress-inducing agents. Role for Nrf1 in gamma-gcs(l) and gss expression in mouse fibroblasts.
AuthorsKwong M, Kan YW, Chan JY
JournalJ Biol Chem
PubMed ID10601325
'Nrf1 is a member of the CNC-basic leucine zipper (CNC-bZIP) family of transcription factors. CNC bZIP factors, together with small Maf proteins, bind as heterodimers to the NF-E2/AP-1 element. Similarity between the NF-E2/AP-1 element and the antioxidant response element identified in a number of promoters of genes involved in detoxification ... More
Simple, fast method of detection apoptosis in lymphoid cells.
'To facilitate the analysis of apoptotic cells, the present study proposes a new quantitative method based on the changes of light scatter properties of lymphoid cells undergoing apoptosis measured with a hematology analyzer. Peripheral blood lymphocytes from 40 chronic B-lymphocytic leukemia samples, five acute T-lymphoblastic leukemia samples, three healthy donors ... More
High-density polymerase-mediated incorporation of fluorochrome-labeled nucleotides.
AuthorsRamanathan A, Pape L, Schwartz DC
JournalAnal Biochem
PubMed ID15649370
'DNA-polymerase-mediated incorporation of different fluorochrome-labeled nucleotides (FdNTPs) was investigated with the goals of optimizing the high-density labeling of probes and exploring DNA sequencing strategies that rely on the controlled, sequential addition of such compounds. By systematically evaluating variables--including polymerase type, buffer conditions, and fluorochrome chemistries--a rational strategy for the sequential ... More
Comparative genomic hybridization with lissamine- and fluorescein-labeled nucleotides.
AuthorsRosenberg C, van Gijlswijk RP, Vos CB, Wiegant J, Cornelisse CJ, Tanke HJ, Raap AK
JournalCytometry
PubMed ID9701403
'Biotin deoxyuridine 5''-triphosphate (dUTP) and digoxigenin dUTP are the labels most commonly used in comparative genomic hybridization (CGH). The relative infrequent use of direct fluorochrome-labeled nucleotides in CGH is related to the lower sensitivity they provide. Here we report the evaluation of two fluorochrome-conjugated nucleotides that have not been previously ... More
Quantitation of fluorescent nucleotide incorporation by capillary gel electrophoresis and laser-induced fluorescence detection.
AuthorsCruickshank KA
JournalAnal Biochem
PubMed ID10094771
'A method has been developed by which enzymatically incorporated fluorophore-labeled nucleotide sites in nucleic acid can be quantitated by degradation of nanogram quantities of DNA followed by capillary gel electrophoretic analysis with fluorescence detection. In this way the differing relative labeling densities achieved using either C5-substituted dUTP''s or N4-substituted dCTP''s ... More
Oxygen stress: a regulator of apoptosis in yeast.
AuthorsMadeo F, Fröhlich E, Ligr M, Grey M, Sigrist SJ, Wolf DH, Fröhlich KU
JournalJ Cell Biol
PubMed ID10330404
Oxygen radicals are important components of metazoan apoptosis. We have found that apoptosis can be induced in the yeast Saccharomyces cerevisiae by depletion of glutathione or by low external doses of H2O2. Cycloheximide prevents apoptotic death revealing active participation of the cell. Yeast can also be triggered into apoptosis by ... More
Cyclosporin inhibition of apoptosis induced by mitochondrial complex I toxins.
AuthorsSeaton TA, Cooper JM, Schapira AH
JournalBrain Res
PubMed ID9795106
The cause of dopaminergic cell death in Parkinson's disease (PD) remains unknown, but may involve oxidative stress and mitochondrial complex I deficiency. Opening of the permeability transition pore and disruption of the mitochondrial transmembrane potential are known to be common events in the apoptotic pathway. Cyclosporin A and its non-immunosuppressant ... More
Apoptosis occurs more frequently in intraductal carcinoma than in infiltrating duct carcinoma of human breast cancer and correlates with altered p53 expression: detected by terminal-deoxynucleotidyl-transferase-mediated dUTP-FITC nick end labelling (TUNEL).
AuthorsHarn HJ, Shen KL, Yueh KC, Ho LI, Yu JC, Chiu SC, Lee WH
JournalHistopathology
PubMed ID9447384
AIMS: We examined the relationship between apoptosis and three different major stages of human breast carcinoma: intraductal carcinoma (DCIS), infiltrating duct carcinoma (IDC) and metastatic carcinoma in lymph nodes. We also determined the correlation between apoptosis and oestrogen receptor (ER), progesterone receptor (PR) and p53. METHODS AND RESULTS: The study ... More
Diffusion of macromolecules in agarose gels: comparison of linear and globular configurations.
AuthorsPluen A, Netti PA, Jain RK, Berk DA
JournalBiophys J
PubMed ID10388779
The diffusion coefficients (D) of different types of macromolecules (proteins, dextrans, polymer beads, and DNA) were measured by fluorescence recovery after photobleaching (FRAP) both in solution and in 2% agarose gels to compare transport properties of these macromolecules. Diffusion measurements were conducted with concentrations low enough to avoid macromolecular interactions. ... More
Direct imaging of DNA in living cells reveals the dynamics of chromosome formation.
AuthorsManders EM, Kimura H, Cook PR
JournalJ Cell Biol
PubMed ID10085283
Individual chromosomes are not directly visible within the interphase nuclei of most somatic cells; they can only be seen during mitosis. We have developed a method that allows DNA strands to be observed directly in living cells, and we use it to analyze how mitotic chromosomes form. A fluorescent analogue ... More
Evaluation of camera requirements for comparative genomic hybridization.
AuthorsTirkkonen M, Karhu R, Kallioniemi O, Isola J
JournalCytometry
PubMed ID8946148
Comparative genomic hybridization (CGH) is based on quantitative digital image analysis of fluorescence intensities from metaphase chromosomes. High-quality CCD cameras are commonly used for image acquisition, but the minimal requirements of CCD cameras have not been determined. We first evaluated minimal camera requirements by artificially reducing spatial and dynamic resolution ... More
Incorporation of reporter-labeled nucleotides by DNA polymerases.
AuthorsAnderson JP, Angerer B, Loeb LA
JournalBiotechniques
PubMed ID15727132
The incorporation of fluorescently labeled nucleotides into DNA by DNA polymerases has been used extensively for tagging genes and for labeling DNA. However, we lack studies comparing polymerase efficiencies for incorporating different fluorescently labeled nucleotides. We analyzed the incorporation of fluorescent deoxynucleoside triphosphates by 10 different DNA polymerases, representing a ... More
Analysis of apoptosis during hair follicle regression (catagen)
AuthorsLindner G, Botchkarev VA, Botchkareva NV, Ling G, van der Veen C, Paus R
JournalAm J Pathol
PubMed ID9403711
Keratinocyte apoptosis is a central element in the regulation of hair follicle regression (catagen), yet the exact location and the control of follicular keratinocyte apoptosis remain obscure. To generate an "apoptomap" of the hair follicle, we have studied selected apoptosis-associated parameters in the C57BL/6 mouse model for hair research during ... More
An evaluation of a new series of fluorescent dUTPs for fluorescence in situ hybridization.
AuthorsWiegant J, Verwoerd N, Mascheretti S, Bolk M, Tanke HJ, Raap AK
JournalJ Histochem Cytochem
PubMed ID8627009
Synthesis of fluorochrome-modified deoxyribonucleotides has been carried out mostly by linking the fluorochrome molecule to the C-5 position of dUTP via an allylamine spacer, similar to the modification of allylamine-dUTP with the haptens biotin and digoxigenin. Recently, a new series of fluorescent nucleotides has been prepared by using an alkynyl ... More
Single-step procedure for labeling DNA strand breaks with fluorescein- or BODIPY-conjugated deoxynucleotides: detection of apoptosis and bromodeoxyuridine incorporation.
AuthorsLi X, Traganos F, Melamed MR, Darzynkiewicz Z
JournalCytometry
PubMed ID7664627
The methods of in situ labeling of DNA strand breaks have been used to identify apoptotic cells and/or DNA replicating cells. While discrimination of apoptotic cells is based on the inherent presence of numerous DNA strand breaks in their chromatin, DNA proliferating cells can be discriminated by the selective DNA ... More
The apoptosis-necrosis paradox. Apoptogenic proteases activated after mitochondrial permeability transition determine the mode of cell death.
AuthorsHirsch T, Marchetti P, Susin SA, Dallaporta B, Zamzami N, Marzo I, Geuskens M, Kroemer G
JournalOncogene
PubMed ID9380409
Mitochondrial alterations including permeability transition (PT) constitute critical events of the apoptotic cascade and are under the control of Bcl-2 related gene products. Here we show that induction of PT is sufficient to activate CPP32-like proteases with DEVDase activity and the associated cleavage of the nuclear DEVDase substrate poly(ADP-ribose) polymerase ... More
Calcium, ATP and nuclear pore channel gating.
AuthorsBustamante JO, Michelette ER, Geibel JP, Dean DA, Hanover JA, McDonnell TJ
JournalPflugers Arch
PubMed ID10678739
Nuclear envelope (NE) cisternal Ca2+ and cytosolic ATP are required for nuclear-pore-complex-(NPC-) mediated transport of DNAs, RNAs, transcription factors and other large molecules. Isolated cardiomyocyte nuclei, capable of macromolecular transport (MMT), have intrinsic NPC ion channel behavior. The large ion conductance (gamma) activity of the NPC channel (NPCC) is blocked ... More
Inhibition of Fas/Fas ligand-mediated apoptotic cell death of lymphocytes in vitro by circulating anti-Fas ligand autoantibodies in patients with systemic lupus erythematosus.
AuthorsSuzuki N, Ichino M, Mihara S, Kaneko S, Sakane T
JournalArthritis Rheum
PubMed ID9485093
OBJECTIVE: The Fas/Fas ligand (FasL) system has been assigned a pivotal role in the establishment and maintenance of peripheral tolerance, and mice having defects in the Fas/FasL system are known to develop lupus-like symptoms. However, it remains unclear whether the Fas/FasL system is involved in the pathogenesis of systemic lupus ... More
Increase in cytosolic Ca2+ levels through the activation of non-selective cation channels induced by oxidative stress causes mitochondrial depolarization leading to apoptosis-like death in Leishmania donovani promastigotes.
AuthorsMukherjee SB, Das M, Sudhandiran G, Shaha C
JournalJ Biol Chem
PubMed ID11983701
Reactive oxygen species are important regulators of protozoal infection. Promastigotes of Leishmania donovani, the causative agent of Kala-azar, undergo an apoptosis-like death upon exposure to H2O2. The present study shows that upon activation of death response by H2O2, a dose- and time-dependent loss of mitochondrial membrane potential occurs. This loss ... More
Cell death in avian tibial dyschondroplasia.
AuthorsRath NC, Huff WE, Bayyari GR, Balog JM
JournalAvian Dis
PubMed ID9533083
Tibial dyschondroplasia (TD) is a local defect of growth plates in fast-growing poultry where the transitional zone cartilage fails to resorb and persists as an avascular plug that prevents endochondral bone formation. We compared the differences in the cartilages from normal and TD-affected growth plates using the reduction of MTS ... More
Effect of anti-neutrophil cytoplasmic antibodies on proteinase 3-induced apoptosis of human endothelial cells.
AuthorsTaekema-Roelvink ME, van Kooten C, Janssens MC, Heemskerk E, Daha MR
JournalScand J Immunol
PubMed ID9714408
Wegener's granulomatosis is characterized by crescentic necrotizing glomerulonephritis and systemic vasculitis. Both proteinase 3 (PR3) and anti-neutrophil cytoplasmic antibodies (ANCA), directed against this enzyme, are thought to play a pathogenic role. PR3 has been shown to cause detachment and cytolysis of human umbilical vein endothelial cells (HUVEC) in vitro and ... More