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A contribution of B cells and autoantibodies has been demonstrated in MS leading to interest in the use of such autoantibodies as diagnostic or prognostic markers and as a basis for immunomodulatory therapy. ELISA and Western blotting fail to detect reactivity against epitopes displayed by native antigens expressed on myelin sheats. We describe a cell-based assay that specifi cally identifi es serum antibodies directed against myelin autoantigens: MBP, PLP and MOG. The method detects antibody binding to recombinant antigens in their native conformation on MBP, PLP or MOG transfected mammalian (hamster ovary) cells. 36 patients with relapsing-remitting MS diagnosed according to criteria of Mc Donald were recruited. Age 38.2 and duration of the disease 7.1. Serum anti-MBP, anti-PLP and anti MOG IgG autoantibodies were detected in MS patients and 35 healthy donors by FACS analysis. Compared with healthy controls the titers of IgG autoantibodies directed against membrane-bound recombinant myelin antigens were most signifi cantly increased for PLP, no quite signifi cant for MBP and not signifi cant for MOG. The titers of anti-MBP antibodies were low in contrast to high titers of anti-MOG antibodies in both groups suggesting a nonspecifi c binding. The cell-based assay detection of autoantibodies directed against recombinant myelin antigens could be a useful tool providing the serological markers in diagnosis and progression of MS. Indeed, it could allow to obtain molecular characteristics of disease in each patient in term of an antibody response against certain myelin and non-myelin antigens. We have shown that in RRMS patients elevated level of serum antibodies against PLP is signifi cant, what should be considered in search for specific immunomodulatory therapy in MS.
There are some hypotheses that oxidative damage to DNA secondary to infl ammation may contribute to irreversible alterations in MS plaques. To test this assumption, we have estimated the level of a DNA oxidative marker, the level of a purine oxidation product, the 8-oxo-2-deoxyguanosine (8-oxo-2dG) in lymphocytes of MS patients. Material and methods: Peripheral blood was collected from 28 patients with clinically defi nite MS aged from 19 to 46 years. The duration of MS was 4.2 ± 3.1 years. The mean of EDSS was 2.0 ± 0.8. In MRI study 14 cases were gadolinium positive and 14 negative. Thirty healthy volunteers make up the control group. DNA was isolated from peripheral blood lymphocytes and then hydrolyzed to nucleosides using P1 nuclease. In order to determine 8-oxo-2dG level, the nucleoside mixture was applied to the HPLC/ UV system, coupled to an electrochemical detector. Results and discussion: The mean level of 8-oxo 2dG in MS patients was 19.6 ± 35.1 and, compared to that established for control subjects (12.3 ± 7.2), showed no statistically signifi cant differences. The comparison of 8-oxo-2dG in subgroups of patients divided according to duration of the disease showed the higher number of cases with DNA damage in patients of the subgroup with shorter duration of the disease. The mean level of 8-oxo-2dG in gadolinium positive MS cases was 17.3 ± 13.1 and in gadolinium negative ones it was 9.6 ± 4.0. The difference was signifi cant. Conclusions: (1) Oxidative damage to DNA is not a general feature in MS patients but it may frequently appear in the early period of the disease; (2) The higher level of oxidative marker of DNA damage in MS, noted in active period of the disease, testifi es to the relationship between the studied variable and MS process.
The question as to the role of immunological mechanisms in neuronal death of extrapyramidal cell systems in Parkinson`s disease is till now not fully resolved. One of the approaches includes an examination of circulating blood cells. In our studies consisting of 24 patients the peripheral blood was studied before and after medication with L-DOPA compounds. Patients with Parkinson`s disease demonstrated an increase of lymphocyte Cd95/CD3 as well as a considerable number of cells dead by apoptotic processes. After treatment with L-DOPA both the percentage of CD95/CD3, acknowledged as an antigen marker characteristic for apoptotic cells as well as the number of cells dead by apoptotic processes were decreased. These findings thus indicate that levodopa treatment in Parkinson`s disease has an impact on apoptotic processes in this instance, and this should be taken into consideration as a positive event in the pathomechanism effected by this treatment.
Introduction: The scientists look for etiopatogenic factors of multiple sclerosis among viruses and bacteria. Genetic and immunologic agents are being taken into consideration. It is more than likely that there are numerous genes that take part in a development of the disease. The influence of various factors at the same time should be considered on account of possible multifactorial etiology of the disease. Aim and material: The aim of the studies was to analyse of MSRV pol, gag, env sequences, TCRB genes and karyotypes in patients with multiple sclerosis. The experimental material was peripheral blood lymphocytes from 130 MS patients and 50 healthy individuals. Methods: Classical (GTG) and molecular cytogenetic techniques (FISH) were used in the experiments. Results: 1) MSRV pol, gag and env sequences were found in both MS patients and controls. 2) The copy number of MSRV sequences was significantly greater in MS patients than in normal individuals. 3) The number of spontaneous micronuclei was significantly greater in MS patients compared to control. 4) Various chromosomal aberrations including translocations between chromosomes 7 and 14 were observed in patients with MS. 5) Translocation of constant and variable TCRB regions, deletion of constant TCRB region at 7 chromosome, duplication of constant TCRB region at chromosome 10, amplification of constant and variable TCRB regions in MS patients with aberrant chromosomes 7 and 14 were also found. Conclusions: 1) Evident difference in MSRV pol, gag and env copy number between MS patients and control suggests that MSRV may play some role in the etiology of multiple sclerosis (latent viral infection). 2) The presence of chromosome aberrations and high amount of micronuclei in MS patients shows that the instability in MS genome often occurs.
A small number (1-5%) of Alzheimer's disease (AD) cases associated with the early-onset form of the disease (EOAD) appears to be transmitted as a pure genetic, autosomal dominant trait. To date, three genes responsible for familial EOAD have been identified in the human genome: amyloid precursor protein (APP), presenilin 1 (PS1), and presenilin 2 (PS2). Mutations in these genes account for a significant frac­tion (18 to 50%) of familial cases of early onset AD. The mutations affect APP pro­cessing causing increased production of the toxic Aβ42 peptide. According to the "amyloid cascade hypothesis", aggregation of the Aβ42 peptide in brain is a primary event in AD pathogenesis. In our study of twenty AD patients with a positive family history of dementia, 15% (3 of 20) of the cases could be explained by coding sequence mutations in the PS1 gene. Although a frequency of PS1 mutations is less than 2% in the whole population of AD patients, their detection has a significant diagnostic value for both genetic counseling and treatment in families with AD.
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