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In physiological conditions proteolytic remodeling of extracellular matrix (ECM) participates in the stem cells development. Signal derived from ECM may activate specifi c intracellular signaling pathways which involve focal adhesion kinase (FAK), a key component responsible for the fl ow of information to the cell. In our study we checked if there is a spatial and temporal relationship between activity of two matrix metalloproteinases (MMP-2 and MMP-9) and neural stem-cell development in gerbil hippocampus 1, 2 and 4 weeks after short-term global ischemia. Furthermore we also investigate the activation of FAK. Our results show that after the ischemia there is a signifi cant increase in the number of proliferating cells in DG which correlates with the activation of MMPs. Whereas in the damaged CA1 pyramidal layer MMP-2 and -9 activity decreases as a result of delayed death of neurons. In the same region we have also noticed a few immature neurons (NF-200 positive) 2 and 4 weeks after reperfusion, however they did not attain maturity. On the other hand we have noticed higher MMPs activity in the astrocytes surrounding the CA1 region. The diverse localization of MMP-2 and -9 activity in both cell types may be due to their different functions – fi rstly with the proliferation of newborn cells and secondly with the creation of the environment while the formation of post injury glial scar. Supported by MSRHE grants: 1266/P01/2006/31
Accumulating evidence indicate that global cerebral ischemia enhanced neurogenesis in adult brain. The mechanism(s) responsible for the arising a new neurons from progenitor cells are poorly understood. Recent in vitro studies indicate the involvement of metalloproteinases (MMPs) in the regulation of proliferation and differentiation of neural progenitor cells, by providing an environment, which is instructive and/or permissive to stem cells activation. To elucidate if MMPs participate in neurogenesis-associated processes after ischemic insult in vivo, we aimed to establish spatial and temporal relationships between neural stem cells development and the modulations of MMPs enzymatic activity in the adult hippocampal dentate gyrus as well as in the ischemia damaged CA1 sector. Our results show that postischemic acceleration of progenitor proliferation in dentate gyrus is accompanied by the increase of MMPs activity. It may indicate that activation of MMPs is likely to be involved in neurogenesis-associated processes. Contrary, the endogenous neurogenesis in the ischemia damaged CA1 pyramidal layer seems to be rather elusive. Despite the appearance of BrdUpositive cells and the newborn neurons they did not attain maturity. Simultaneously, the activity of MMPs in this area was markedly reduced. The increase of MMPs activity seen in the astrocytes in the neighboring structures ñ stratum oriens and stratum radiatum, probably plays a role in delayed tissue remodeling and delayed repair processes. In conclusion, our results show that MMPs may, at least in part, contribute to ischemia-induced neurogenesis in the dentate gyrus of the adult brain, along with other previously reported factors. This work was supported by MSHE grant 1266/P01/2006/31
Stroke resulting from cerebral ischemia or haemorrhage is a common cause of the death of neuronal cells and neurological dysfunction in humans. Finding therapeutics to improve outcome from brain ischemic injury has proved to be challenging. The efficacy of neuroprotective compounds identified in experimental brain ischemia models thus far have failed to successfully translate in clinical human trials. Recent experimental evidence indicates that inhibition of zinc-dependent histone deacetylases can protect neuronal and oligodendroglial cells from the damaging effects of ischemic insult, which may contribute to improved functional outcome. In this review we briefly highlight the current data supporting a beneficial role of histone deacetylation in experimental brain ischemia. We also discuss the molecular mechanism of neuroprotection.
The bioaccumulation ability of radionuclides 51Cr, 54Mn, 57Co, 60Co, 65Zn, 85Sr, 109Cd, 110mAg, 113Sn, 137Cs and 241Am in two red algae species from the southern Baltic Sea – Polysiphonia fucoides and Furcellaria lumbricalis – was determined under laboratory conditions. P. fucoides demonstrated better bioaccumulative properties towards most of the investigated radionuclides. As a result, P. fucoides can be recommended as a good bioindicator of radioactive environmental pollution. The bioaccumulation of radionuclides in F. lumbricalis was studied during an extended laboratory experiment. The initial extensive uptake of radioisotopes was followed by the rapid removal of cations; in general, concentrations tended to decrease with time. 137Cs displayed a different behaviour, its concentration in the algae increasing over time mainly due to its large ion radius; this is a factor that could be responsible for the stronger mechanical and chemical bonding of Cs+ and that could hamper the movement of ions in both directions.
The discovery of neurogenic response subsequent to brain injuries has led to the hypothesis that the expansion of the pool of endogenous progenitors could augment the regenerative capacity of the damaged areas. However, it occurred that endogenous spontaneous neurogenesis is insufficient for replacing the lost neurons and to achieve global repair, particularly in aging brain. Until today, a great effort has been made attempting to promote "reactive neurogenesis" more successful. It was found that small chemical molecules exert stimulation of neurogenesis and probably might help to induce neuronal endogenous cell replacement in various neurological diseases. In this review we briefly highlight the current data regarding effect of brain ischemia and age-related neurodegenerative diseases on neural stem cells in situ and potential therapeutic effect of their stimulation.
Brain ischemia resembles other brain injuries in producing enhanced neurogenesis in neuroproliferative regions of the rodent brain, including subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) of the dentate gyrus (DG) of the hippocampus. Newly-generated neurons would be incorporated in the hippocampal local circuitry and involved in brain repair. Organotypic culture of hippocampal slices (OHC) provides an alternative model of hippocampus in vivo. Moreover, exposure of the organotypic slices to oxygen and glucose deprivation (OGD) mimics cerebral ischemia. The aim of the present study was to investigate whether deprivation of oxygen and glucose might stimulate cell proliferation and neurogenesis in organotypic hippocampal slice culture. Furthermore, we evaluate whether the activity of matrix metalloproteinases (MMPs) in the OHC parallels the rate of cell proliferation and/or further differentiation. Cell death in the organotypic hippocampal slices was determined with propidium iodide (IP) staining. Stem cells proliferation was detected by using DNA replication marker – 5-Bromo 2-Deoxyuridine (BrdU) followed by immunoreaction with specific antibodies. Newly generated BrdU(+) cells were identified by an analysis of neural, glial and microglial markers expression – NF-200 NeuN, GFAP, ED1, respectively. In order to check the activity and localization of metalloproteinases, MMP-2 and MMP-9, we conducted in situ zymography in conjunction with immunohistochemistry. Exposing rat OHC for 40 min OGD followed by 24h of reoxygenation induces cell death in CA1 area with only negligible damage in DG. At 1 week cell death appears all over the slice in control conditions as well as after OGD. The stimulation of cell proliferation was observed 7 days after OGD exclusively in CA1. At the same time the number of BrdU(+) cells in DG remained on the level characteristic for control cultures. The majority of BrdU positive cells presents expression of microglial specific stain (ED1) pronounced particularly in CA1 at 3 days after OGD. However, some BrdU labeled nuclei were encapsulated by GFAP positive processes especially in CA1 region of the hippocampus (3 and 7 days after OGD). We do not notice coexpression of BrdU-positive cells with NeuN(+) mature neurons. The study suggests that slice cultures do not show neurogenesis for chosen cultivation period. Activation of MMPs was localized mainly in microglial cells and may be associated with their proliferation in situ. Supported by MSHE grant no 0154/B/P01/2010/38
Oligodendrocytes, the cells responsible for myelin formation and maintenance in CNS, are depleted in many acute and chronic conditions. The stem/progenitor cells stimulation or transplantation might be seriously considered as a long hopedfor therapeutic perspective. Better understanding of the mechanism(s) regulating the activation of the cell lineage from the endogenous progenitor reservoir might be helpful. Therefore an efficient source of donor cells for transplantation in humans is being craved for. In this study we show that the application of extracellular matrix component-laminin promotes oligogliogenesis from neural stem-like cells of human cord blood cells (HUCB-NSC). Although oligodendrocytes constitute a minor subpopulation of spontaneously differentiated HUCB-NSC, the manipulation of active compounds regulating the process of cell commitment results in a several fold increase in their number. Thus cells of the HUCB-NSC line could be considered as a potential source of glial cells, fulfilling the suitable candidate criteria for oligodendrocyte replacement therapy.
Celem pracy była ocena przydatności krajowych odmian jabłek i ocena jakości otrzymanych z nich napojów pod kątem przydatności do bezpośredniego spożycia oraz do produkcji napoju typu calvados. Dokonano oceny soków otrzymanych z jabłek oraz zagęszczonego soku jabłkowego (oznaczenie zawartości ekstraktu ogólnego, cukrów, kwasowości ogólnej); określono jakość win owocowych (poprzez oznaczenie mocy, zawartości ekstraktu, cukrów, substancji smakowo-zapachowych wpływających na jakość otrzymanych wyrobów, kwasowości ogólnej wyrażonej jako kwas jabłkowy, ocenę organoleptyczną wyrobów bezpośrednio po zakończeniu fermentacji i po okresie dojrzewania), a także oceniono przydatność cydrów do produkcji „calvadosu” (poprzez destylację i leżakowanie wytypowanych napojów typu cydr, ocenę walorów smakowo-zapachowych otrzymanych napojów, w tym: oznaczenie mocy, zawartości ekstraktu, ubocznych produktów fermentacji alkoholowej, substancji smakowo-zapachowych wpływających na jakość otrzymanych wyrobów, porównawczą ocenę organoleptyczną otrzymanych wyrobów z francuskimi calvadosami). Zastosowane w niniejszej pracy warunki fermentacji (10°C przez 40 dni i 15°C przez 28 dni) doprowadziły do uzyskania napojów typu cydr i calvados ocenianych jako dobre oraz zbliżone w ocenie organoleptycznej do wyrobów francuskich. Fermentacja w temperaturze 22°C (przez 21 dni) i 30°C (przez 12 dni) spowodowała powstanie produktów o słabszej charakterystyce sensorycznej oraz niższych zawartościach alkoholu etylowego. Ocena organoleptyczna cydrów otrzymanych podczas fermentacji spontanicznej wskazywała na niższą akceptację tych wyrobów przez degustatorów. Cydry otrzymane z polskich jabłek oraz zagęszczonych soków jabłkowych są odpowiednie, zarówno do bezpośredniego spożycia, jak i do produkcji calvadosu.
Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are considered the main regulators of the cell microenvironment which governs NSC development. The aim of our study was to investigate whether the selected ECM components (laminin, fi bronectin, collagen) infl uence the neural stem cells proliferation and differentiation. In addition we checked if MMPs are engaged in the above developmental processes. Methods: The cells of HUCB-NSC line were seeded on the ECM components-coated plates. The serum-free medium was applied 48 h before experiments. On the 4th, 8th and 14th day in culture, cell proliferation assay and in situ zymography were performed, followed by the immunocytochemistry with the specifi c neural markers. Results: Our results showed that fi bronectin stimulated cell proliferation as well as MMPs activity most intensively (~20% increase in the 2-week cultured cells). It was also the most potent factor in promoting the cell differentiation, mostly toward neurons. We also checked the involvement of the MMPs in cell development. For this purpose we used MMPs inhibitors: GM6001 and TIMPs as well as inhibitors for serine and furin proteases ñ Pefabloc and DecRVKR-CMK. We found that only inhibitors of MMPs infl uence stem cells developmental processes. GM6001 down-regulated cell proliferation (~30%) and differentiation into neurons (~20%). The decrease of cell proliferation was also observed in the presence of TIMP2. In contrast, TIMP1 accelerated cell divisions. Conclusion: Our results demonstrate that fi bronectin is the potent factor in promoting the cell proliferation, differentiation, and support the participation of MMPs in the mechanism(s) responsible for neural stem cells maturation. Supported by MSRHE grants: 1266/P01/2006/31 and N40101832 /0296
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