The Experts below are selected from a list of 30 Experts worldwide ranked by ideXlab platform
J Nilsson - One of the best experts on this subject based on the ideXlab platform.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis.
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell-death in atherosclerotic plaques with a recent history (< 6 months) of rupture. Atherosclerotic plaques were obtained from patients with symptomatic ipsilateral carotid stenosis > 70% diameter reduction undergoing carotid endarterectomy. In situ tailing and Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and oncosis. The mean number of cells containing fragmented DNA in the plaques was 12.7 +/- 3.5% (n = 15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated from part of plaques, where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation, resulted in multiple ladders of 180-200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by oncosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques, oncosis appears to be a much more common mechanism for cell death.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell death in atherosclerotic plaques with a recent history ( 70% diameter reduction) undergoing carotid endarterectomy. In situ tailing Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and necrosis. The mean number of cell containing fragmented DNA in the plaques was 12.7 ± 3.5% (n=15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated form part of plaques where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation resulted in multiple ladders of 180–200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by necrosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques necrosis appears to be a much more common mechanism for cell death. This may have negative consequences for plaque stability since cell death by necrosis induces a stronger inflammatory reaction than apoptotic cell death.
M Crisby - One of the best experts on this subject based on the ideXlab platform.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis.
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell-death in atherosclerotic plaques with a recent history (< 6 months) of rupture. Atherosclerotic plaques were obtained from patients with symptomatic ipsilateral carotid stenosis > 70% diameter reduction undergoing carotid endarterectomy. In situ tailing and Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and oncosis. The mean number of cells containing fragmented DNA in the plaques was 12.7 +/- 3.5% (n = 15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated from part of plaques, where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation, resulted in multiple ladders of 180-200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by oncosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques, oncosis appears to be a much more common mechanism for cell death.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell death in atherosclerotic plaques with a recent history ( 70% diameter reduction) undergoing carotid endarterectomy. In situ tailing Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and necrosis. The mean number of cell containing fragmented DNA in the plaques was 12.7 ± 3.5% (n=15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated form part of plaques where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation resulted in multiple ladders of 180–200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by necrosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques necrosis appears to be a much more common mechanism for cell death. This may have negative consequences for plaque stability since cell death by necrosis induces a stronger inflammatory reaction than apoptotic cell death.
Gabor Huszar - One of the best experts on this subject based on the ideXlab platform.
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A new media without animal component for sperm cryopreservation: motility and various attributes affecting paternal contribution of sperm
Journal of Assisted Reproduction and Genetics, 2017Co-Authors: Akansha Tiwari, Leyla Sati, Merih Tekcan, William Murk, Jill Stronk, Gabor HuszarAbstract:Purpose Our aim was the development of a safe sperm cryopreservation New Media (NM), composed of consistent and reproducible components devoid of any animal origin, and evaluation of NM in terms of its effect on sperm structure and function as compared to regularly used yolk media (TYM) (Irvine Scientific). Methods We evaluated patient semen samples and cryopreserved them in duplicates in either NM or TYM. The samples were cryopreserved for either a short term of 1 week or long term of 1 month prior to thawing. The parameters investigated include sperm motility via computer-assisted semen analysis (CASA), sperm concentration, and sperm biomarkers that promote paternal contribution of spermatozoa to fertilization including hyaluronic acid binding, chromatin maturity, apoptotic markers, cytoplasmic retention, and sperm DNA integrity. Results As compared to TYM, NM was equally capable of sperm cryopreservation with both short-term and long-term storage in media, and after freeze-thaw and gradient processing of sperm. HA binding of sperm was comparable post thaw in both NM and yolk media. There are also no differences observed between the samples cryopreserved in NM or TYM in terms of their aniline blue staining, CK immunocytochemistry, caspase 3 immunostaining, or DNA Nick Translation. Conclusions NM has the advantage of being xeno-free, yet in preservation of sperm motility and other sperm attributes, the NM is as effective as the TYM.
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Methodology of Aniline Blue Staining of Chromatin and the Assessment of the Associated Nuclear and Cytoplasmic Attributes in Human Sperm
Methods of Molecular Biology, 2012Co-Authors: Leyla Sati, Gabor HuszarAbstract:In this chapter, the laboratory methods for detection of sperm biomarkers that are aimed at identifying arrested sperm development are summarized. These probes include sperm staining with aniline blue for persistent histones, representing a break in the histone-transition protein-protamine sequence, immunocytochemistry with cytoplasmic sperm proteins, highlighting cytoplasmic retention during spermiogenesis, DNA Nick Translation testing for DNA chain fragmentation due to various reasons, for instance low HspA2 chaperone protein levels, and consequential diminished DNA repair. Finally, we briefly provide references on our work on sperm hyaluronan binding, abnormal Tybergerg sperm morphology, and the increased levels of chromosomal aneuploidies in sperm with developmental arrest. A very interesting aspect of the biomarker field is the discovery (Sati et al, Reprod Biomed Online 16:570-579, 2008) that the various nuclear and cytoplasmic defects detected by the biomarkers are related, and may simultaneously occur within the same spermatozoa as evidenced by a combination of biomarkers, such as aniline blue staining (persistent histones) coupled with cytoplasmic retention, DNA fragmentation, Caspase-3, Tygerberg abnormal morphology, and increased levels of chromosomal aneuploidies. We show examples of this >80% overlap in staining patterns within the same spermatozoa.
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Semen Characteristics After Overnight Shipping: Preservation of Sperm Concentrations, HspA2 Ratios, CK Activity, Cytoplasmic Retention, Chromatin Maturity, DNA Integrity, and Sperm Shape
Journal of Andrology, 2004Co-Authors: Gabor Huszar, Ciler Celik-ozenci, Sevil Cayli, Tamas Kovacs, Lynne Vigue, Ertug KovanciAbstract:ABSTRACT: We tested several approaches that can be used topreserve sperm attributes and the objective biochemical markers ofsperm maturity and function for assessment in a remote centralizedlaboratory after overnight shipping of semen samples. Addition ofphenyl-methyl-sulfonyl-fluoride (PMSF) to a final concentration of 20mg/mL semen at 48C has preserved sperm concentrations andHspA2 isoform ratios, even at room temperature, simulating a ship-ping delay in moderate ambient temperatures. Regarding the attri-butes of individual spermatozoa, the patterns of CK-immunocyto-chemistry (demonstrates cytoplasmic retention in diminished-matu-rity spermatozoa); aniline blue staining pattern (tests chromatin ma-turity); sperm shape assessed by both Kruger strict morphology andcomputer assisted morphometry; and sperm DNA integrity, as testedby DNA Nick Translation, all remained unchanged. Thus, the PMSF-48C conditions preserved sperm concentrations and the cytoplasmicand nuclear biomarkers of sperm cellular maturity and function fornext-day analysis. This shipping method will facilitate the early de-tection of subtle changes in semen quality that can affect spermfunction, even when there has been no decline in sperm concentra-tions to signal possible toxic effects. Furthermore, sample preser-vation will enable investigators to evaluate semen for toxicologystudies and for diagnosis of male infertility from remote locations.Home collection of semen should enhance study participation, andsemen assessment in centralized laboratories will address concernsregarding interlaboratory variations and quality control.Key words: Reproductive toxicity, male infertility, cytoplasmic andnuclear biomarkers.J Androl 2004;25:593–604
V Kostulas - One of the best experts on this subject based on the ideXlab platform.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis.
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell-death in atherosclerotic plaques with a recent history (< 6 months) of rupture. Atherosclerotic plaques were obtained from patients with symptomatic ipsilateral carotid stenosis > 70% diameter reduction undergoing carotid endarterectomy. In situ tailing and Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and oncosis. The mean number of cells containing fragmented DNA in the plaques was 12.7 +/- 3.5% (n = 15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated from part of plaques, where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation, resulted in multiple ladders of 180-200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by oncosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques, oncosis appears to be a much more common mechanism for cell death.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell death in atherosclerotic plaques with a recent history ( 70% diameter reduction) undergoing carotid endarterectomy. In situ tailing Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and necrosis. The mean number of cell containing fragmented DNA in the plaques was 12.7 ± 3.5% (n=15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated form part of plaques where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation resulted in multiple ladders of 180–200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by necrosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques necrosis appears to be a much more common mechanism for cell death. This may have negative consequences for plaque stability since cell death by necrosis induces a stronger inflammatory reaction than apoptotic cell death.
B Kallin - One of the best experts on this subject based on the ideXlab platform.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis.
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell-death in atherosclerotic plaques with a recent history (< 6 months) of rupture. Atherosclerotic plaques were obtained from patients with symptomatic ipsilateral carotid stenosis > 70% diameter reduction undergoing carotid endarterectomy. In situ tailing and Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and oncosis. The mean number of cells containing fragmented DNA in the plaques was 12.7 +/- 3.5% (n = 15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated from part of plaques, where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation, resulted in multiple ladders of 180-200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by oncosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques, oncosis appears to be a much more common mechanism for cell death.
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Cell death in human atherosclerotic plaques involves both oncosis and apoptosis
Atherosclerosis, 1997Co-Authors: M Crisby, B Kallin, J Thyberg, B Zhivotovsky, S Orrenius, V Kostulas, J NilssonAbstract:The aim of the present study was to analyze the frequency and mechanism of cell death in atherosclerotic plaques with a recent history ( 70% diameter reduction) undergoing carotid endarterectomy. In situ tailing Nick Translation of fragmented DNA, agarose gel electrophoresis of plaque DNA and electron microscopy were used to identify cell death by apoptosis (programmed cell death) and necrosis. The mean number of cell containing fragmented DNA in the plaques was 12.7 ± 3.5% (n=15). Focal accumulations of cells with DNA fragmentation occurred in the fibrous cap, at sites of rupture, close to lipid deposits and necrosis and was always accompanied by the presence of inflammatory cells. Electrophoretic separation of DNA isolated form part of plaques where the presence of DNA fragmentation had previously been demonstrated by in situ DNA Nick Translation resulted in multiple ladders of 180–200 base pairs characteristic of apoptosis. Electron microscopic analysis revealed presence of cells with morphological signs of degeneration in a frequency even higher than that found by in situ Nick Translation. Some of these cells had a characteristic apoptotic appearance with condensed chromatin and cytoplasm, but the large majority of the cells had an ultrastructure typical for cells undergoing cell death by necrosis with membrane disruption and swollen, disintegrating organelles. Thus, although apoptosis clearly takes place in atherosclerotic plaques necrosis appears to be a much more common mechanism for cell death. This may have negative consequences for plaque stability since cell death by necrosis induces a stronger inflammatory reaction than apoptotic cell death.