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To assess the dependence on age of the expression of apoptosis regulatory proteins in the human semitendinosus muscle, we measured the expression levels of several apoptosis-related genes, including apoptosis-inducing factor (AIF), Bax, Bcl-2, caspase-3 and heat shock protein 70 (HSP70), using RT-PCR, immunohistochemistry and TUNEL assays. We found that the DNA fragmentation was proportional to the age of the tissues sample donors. The expression levels of AIF were significantly elevated (by 10 to 25%) in semitendinosus tissue samples from older individuals, but the Bax, Bcl-2, caspase-3 and HSP 70 levels remained almost constant. This data suggests that the morphological and functional changes observed in aged human semitendinosus muscle correlates with the apoptosis of muscle cells through the induction of AIF.
The temperature dependence of the activity and structure of the enzyme carbonic anhydrase was studied. The Arrhenius plot shows a jump which is seen usually in proteins with more than one subunit or with one subunit but more than one domain. Since carbonic anhydrase has only one subunit with one domain, the fine conformational changes of the protein motifs could only be detected through circular dichroism polarimetry. It seems that the jump in Arrhenius plot is a result of some slight structural changes in the secondary and tertiary structures of the enzyme.
Ophthalmic diseases, especially retinal degeneration belong to prominent causes of disability in developed countries. Thus, special attention has been focused on research aimed on establishing new protocols of efficient stem cell (SC)-based therapy of these disorders. The aim of this study was to determine and optimize a new strategy of SC-based therapy of selectively damaged retina after sodium iodate (NaIO3) administration in C57BL/6J mice. First, we sought to assess the regenerative mechanisms triggered after acute chemical injury of retinal pigment epithelium and neurosensory retina induced by NaIO3, in mice. The intravenous injection of NaIO3 provides a useful model for the study of retinal degeneration since it mimics some retinal degenerative diseases in humans, e.g., gyrate atrophy, retinitis pigmentosa or age-related macular degeneration. We evaluated the kinetics of morphological and functional changes within mice retinas injured with NaIO3 via: (1) morphological study; (2) evaluation of expression of selected neurotrophins (NTs) in injured retina; (3) visualization of proliferating and apoptotic cells; (4) electrophysiology. Our findings revealed that massive destruction of the tissue was associated with irreversible retinal dysfunction, whereas moderate retinal injury triggered regenerative mechanisms that restore bioelectrical function of the damaged retina. Next, we performed intravitreal transplantation of murine GFP+Lin- cells on the 1st day since NaIO3 administration. We analyzed number and localization of intravitreally injected GFP+Lin- cells within recipients’ retinas as well as the retinal functional changes (electroretinography). By employing stem/progenitor cell-based therapy we achieved noticeable improvement in retinal function, particularly in the case of only partial primary destruction of retinal tissue. Furthermore we investigated the neuroprotective effects of different NTs, administered intravitreally via genetically modified SCs into degenerating retinal tissue. The synthetic viral vectors based on lentivirus (LVs) backbone was used to deliver NT genes into mesenchymal stem cells isolated from bone marrow. Then, we conducted multipart analysis based on functional tests, e.g., electroretinography as well as histological, immunohistochemical, morphometric, and molecular biology studies. We found that specific, exogenously administered NTs, such as NT4/5 could effectively stimulate photoreceptor survival in the degenerating retinas. Our findings reveal that the proposed therapeutic strategy could be recommended as adjuvant therapy supporting endogenous regeneration of acute retinal damage. However, further more extensive studies are needed before the introduction of this kind of therapy into patients.
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Pathophysiology of ageing

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Ageing is characterized by a gradual decline in organ functional reserves which reduces the ability to maintain homeostasis under conditions of stress. Introduction of cell culture and molecular biology techniques has provided new experimental tools for the analysis of ageing at the molecular level. During ageing progressive degeneration of cells and loss of regenerative capacity are enhanced and with time the alterations caused by them ultimately lead to death. In this paper the current knowledge of the mechanisms of ageing is summarized.
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