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EN
Biological studies rely on measurements performed on large populations, which allow the statistic evaluation of gathered data. On the other hand, the need to precisely localize observed processes and the facts occurring in the microscale results in the need of high resolution qualitative imaging. These two approaches are complementary and without anyone of them no valuable experimental data can be gathered from studies of fluorescent dyes. Both approaches are based on common origin of laser-induced fluorescence, but due to the method of analysis they split into the quantitative (laser scanning cytometry) and qualitative (confocal microscopy) fields of research.
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EN
In this study the contribution of the ERK1/2 pathway to sphingosine-induced death and morphological changes of the actin cytoskeleton in glioma C6 cells was investigated. Surprisingly, the level of ERK1/2 phosphorylation does not change after incubation of cells with sphingosine. Despite this, sphingosine induces rounding and detachment of cells without formation of apoptotic bodies. To shed light on this process, a specific inhibitor of ERK1/2 phosphorylation, U0126, was used. Cells incubated simultaneously with sphingosine and U0126 not only detached, but also exhibited formation of apoptotic-like blebs. These data suggest that during sphingosine-induced glioma C6 cell death apoptotic blebbing is dependent on ERK1/2 signalling and occurs only when ERK1/2 activity is decreased or abolished.
EN
Apoptosis, or programmed cell death (PCD), involves a complex network of biochemical pathways that normally ensure a homeostatic balance between cellular proliferation and turnover in nearly all tissues. Apoptosis is essential for the body, as its deregulation can lead to several diseases. It plays a major role in a variety of physiological events, including embryonic development, tissue renewal, hormone-induced tissue atrophy, removal of inflammatory cells, and the evolution of granulation tissue into scar tissue. It also has an essential role in wound repair. The various cellular and biochemical mechanisms involved in apoptosis are not fully understood. However, there are two major pathways, the extrinsic pathway (receptor-mediated apoptotic pathway) and the intrinsic pathway (mitochondria-mediated apoptotic pathway), which are both well established. The key component in both is the activation of the caspase cascade. Caspases belong to the family of proteases that ultimately, by cleaving a set of proteins, cause disassembly of the cell. Although the caspase-mediated proteolytic cascade represents a central point in the apoptotic response, its initiation is tightly regulated by a variety of other factors. Among them, Bcl-2 family proteins, TNF and p53 play pivotal roles in the regulation of caspase activation and in the regulation of apoptosis. This review summarizes the established concepts in apoptosis as a physiological cell suicide program, highlighting the recent and significant advances in its study.
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PL
Zanieczyszczenia gleby metalami ciężkimi mają toksyczne działanie na rośliny, zwierzęta oraz człowieka. Metalofity, rośliny odporne na metale ciężkie, kolonizujące tereny metalonośne, są wykorzystywane do fitoremediacji, czyli oczyszczania gleb z metali ciężkich. Wykorzystanie roślinnych kultur komórkowych do badań nad toksycznością metali i tolerancją komórek na ich działanie jest stosunkowo nową techniką. W pracy zostały przedstawione możliwości wykorzystania roślinnych kultur zawiesinowych w badaniach nad wpływem metali ciężkich na metabolizm komórek oraz metody oszacowania ich toksycznego wpływu. Zaprezentowane zostały techniki otrzymywania kultur zawiesinowych, oceny żywotności komórek, akumulacji metali ciężkich w komórkach. W ocenie toksyczności metali stosuje się także badania nad programowaną śmiercią komórki (PCD), co pozwala oszacować reakcję komórek na ich wysokie stężenia. Zostały przedyskutowane mechanizmy tolerancji komórek na metale ciężkie. Kultury zawiesinowe są dobrym modelem do badań tolerancji na metale, ponieważ pozwalają zbadać ich wpływ na pojedyncze komórki w jednolitych, stałych warunkach.
EN
Soil pollutants exert toxic effects on plants, animals and humans. Metallophytes, plants tolerant to heavy metals colonizing polluted areas, are being used to phytoremediation - cleaning up soil contaminated with heavy metals. The use of plant cells in vitro cultures to study heavy metal toxicity and tolerance is a relatively new approach in research of metal toxicity. In this paper the usefulness of plant suspension cultures to study the impact of heavy metals on cells is presented alongside with the methods of obtaining suspension cultures, evaluation of cell viability, metal accumulation and detection of programmed cell death (PCD). The mechanisms by which cells of plant species tolerant to heavy metals develop resistance to metal toxicity are discussed. Cell suspension cultures appear to be a good model to study tolerance to heavy metals because they allow to estimate metal impact to a single cell in stable uniform conditions.
EN
The use of soybean in human and animal nutrition is limited because of high content of bioactive compounds: enzyme inhibitors, polyphenols, goitrogens, phytates, saponins, sugars, and agglutinins. The damage of intestinal mucosa structure was previously observed in animals fed soybean supplemented diets. Hence, the objectives of the presented study were to compare intensity of epithelium remodeling processes in different intestinal segments, and to evaluate the influence of the 1% of soybean dietary supplementation on the processes in intestinal mucosa. The experiment was performed on 30 Wistar rats fed AIN-93 based diets. Animals were divided randomly into three groups: control (CTRL), with 1% of raw soybean (RS) and with 1% of soaked and boiled soybean (BS). The samples of: duodenum (DUO), proximal jejunum (PROX), mid-jejunum (MID), distal-jejunum (DIST) and ileum (ILE) were collected. The following processes in these samples were evaluated: mitosis (Ki-67), apoptosis (Cpp32), autophagy (MAP I LC3) and DNA damage (p53). Present data show that modification of soybean by soaking and subsequent boiling markedly influences the enterocyte turnover in the small intestine mucosa. Increased mitotic ratio in the intestine of rats fed with boiled soybean masks the negative effects of soybean on the small intestine structure.
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