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  • 1
    Abstract: Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, but the underlying mechanism(s) remain(s) to be elucidated. We examined ionomycin-induced cell death in LCLC 103H cells, derived from a human large cell lung carcinoma. We detected hallmarks of apoptosis such as membrane blebbing, nuclear condensation, DNA ladder formation, caspase activation, and poly-(ADP-ribose)polymerase cleavage. Apoptosis was prevented by preincubation of the cells with the calpain inhibitor acetyl-calpastatin 27-peptide and the caspase inhibitor Z-DEVD-fmk, implicating both the calpains and caspases in the apoptotic process. The apoptotic events correlated in a calpastatin-inhibitable manner with Bid and Bcl-2 decrease and with activation of caspases-9, -3, and -7. In vitro both ubiquitous calpains cleaved recombinant Bcl-2, Bid, and Bcl-x(L) at single sites truncating their N-terminal regions. Binding studies revealed diminished interactions of calpain-truncated Bcl-2 and Bid with immobilized intact Bcl-2 family proteins. Moreover, calpain-cleaved Bcl-2 and Bid induced cytochrome c release from isolated mitochondria. We conclude that ionomycin-induced calpain activation promotes decrease of Bcl-2 proteins thereby triggering the intrinsic apoptotic pathway.
    Type of Publication: Journal article published
    PubMed ID: 12000759
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  • 2
    ISSN: 0920-9964
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have investigated T-cell antigen receptor constant β chain genes (Tcr C β) and immunoglobulin (Ig) heavy chain switch region genes of HLA-DR-typed patients with membranous nephropathy (MN) employing DNA restriction fragment length polymorphism (RFLP) analysis. When a Tcr C β probe in conjunction with the restriction endonuclease BgI II was used, a significant increase in the frequency of a 10.0;9.2 kb heterozygous RFLP phenotype was found in MN (75.0 % versus 42.1 in controls; P=0.002). When Sst I-restricted DNA from MN patients was hybridized with a DNA probe homologous to the switch region flanking the Ig C µ heavy chain gene (S µ), there was a significant decrease in the frequency of the 2.1; 2.6 kb heterozygous RFLP phenotype in MN (24.0% versus 54.6% in controls; P=0.004). These results suggest that Tcr beta and Ig heavy chain loci, as well as HLA antigens, may be important in the pathogenesis of MN.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Narcolepsy has a 98% association with the DR2-Dw2/DQw1 haplotype. To establish if a disease-specific allele is present in narcolepsy, a cDNA library was made from a B-cell line from a DR2,4/DQw1,3 narcoleptic. Clones encoding the two expressed DR2 β chains, along with DQw1 α and β chains, were isolated and completely sequenced. The coding regions of these four genes were similar to published nucleotide and protein sequences from corresponding healthy controls, with some minor exceptions. The 3′ untranslated region of one of the DR2 β genes in the narcoleptic was extended by 42 bp. Complete sequences were not available for DQw1.2 α or β from healthy individuals, but first domain nucleotide sequences showed only a single nonproductive difference in DQα. Partial protein sequences of both DQ α and β from published data were identical. Although the effects of minor differences cannot be ruled out completely, it is concluded that there are probably no narcolepsy-specific DR β or DQ α/β sequences, and that the alleles found in narcolepsy are representative of those found in the healthy population.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2277
    Keywords: Antibodies, and loss of renal grafts ; Non-HLA antibodies, loss of renal grafts ; Rejection, multiple, kidneys, novel antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using an endothelial/epithelial hybrid cell line, three different non-HLA antibody types have been identified by flow cytometry in patients who have rapidly rejected multiple renal allografts. These antibodies may be classified as anti-endothelial-monocyte, anti-activated endothelial cell, or anti-epithelial cell.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2277
    Keywords: HLA ; Kidney ; Transplantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Transplantation of kidneys bearing HLA antigens to which recipients have previously been exposed is generally avoided, and such prudence is a well-documented means of preventing early graft loss. Prior exposure and subsequent reactions can, however, take a wide variety of forms, and blanket avoidance may prevent many deserving patients from being transplanted. In our region, operating through a single tissue-typing laboratory, we follow a consistent policy of allowing retransplantation with kidneys bearing previous mismatches, provided no relevant antibody response has been detected. Twenty-one of 34 such transplants remain functioning at time periods ranging from 7 months to 7 years. Four were lost due to rejection within the 1st month, and the remaining 9 functioned for periods ranging from 2 months to 8 years. Three were lost for reasons other than rejection. Our antibody screening policy and our criteria for a negative crossmatch results in the exclusion of two-thirds of all repeat mismatch transplantations. The results indicate that in the remaining third, transplantation can be performed across a repeat mismatch with excellent long-term results, provided our defined crossmatch policy is adhered to strictly.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2277
    Keywords: Pancreas, foetal, immunology ; Foetal pancreas, immunology ; Immunology, foetal pancreas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression of MHC class I and class II antigens by the human foetal pancreas (HFP) during the first trimester is poorly documented. Using immunohistochemical techniques, we analysed 37 HFPs aged 8–13 gestatitional weeks (gw) and compared the results with those of 9 HFPs aged 14–16 gw. In all of the specimens, the ductal cells were class I- and class II-negative. Islets and endothelial cells expressed class I but were class II-. Interstitial class II+ cells included macrophages, B lymphocytes and dendritic-like cells that were negative for macrophage markers. While the frequency of class II+ cells in the HFP remained constant from 8 to 13 gw, a threefold increase was observed from the end of the 13th gw to the 16th gw. In conclusion, the lower density of interstitial class II+ cells in HFPs aged 8–13 gw indicates that immunomodulation is likely to be more successful in this age group.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2277
    Keywords: MHC regulation, human fetal pancreas ; Fetal pancreas, human, MHC regulation ; Gamma-interferon, MHC regulation, human fetal pancreas ; Pancreas, human fetal, MHC regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunological data on the human fetal pancreas (HFP) are mainly confined to its constitutive expression of the MHC antigens. However, cytokines, such as gamma-interferon (g-IFN), released by lymphocytes during immune reactions, can induce or upregulate the expression of MHC products in allografts and alter their immunological behaviour. We investigated the effects of g-IFN on fresh and cultured HFPs aged 9–16 gestational weeks (gw). Following g-IFN stimulation of fresh HFPs, there was class I hyperexpression by the ductal cells, and some of the ductal, endothelial and islet cells also became class II+. Conventional tissue culture (5% CO2 in air at 37°C) reduced the number of interstitial class II+ cells within the HFP after 1 week but was associated with de novo class I expression by some of the ductal cells. Remarkably, the changes in major histocompatibility complex (MHC) antigen expression by the ductal cells occurred earlier and were markedly enhanced when the HFPs were cultured beforehand. The number of interstitial class II+ cells in fresh and cultured HFPs was not influenced by g-IFN. The significance of these observations with regard to clinical HFP transplantation is discussed.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2277
    Keywords: HLA class I and II antibodies, screening ; Antibodies, HLA,screening ; Screening, antibodies, HLA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Screening of potential transplant recipients for antibodies that can cause graft rejection is an essential part of the pre-transplant monitoring carried out by tissue typing laboratories. This is a time-consuming process and the rapid reporting of results is dependent on the maintenance of frozen cell panels. The usual procedure of screening against a panel of random cells takes up to 6 weeks. In this study we have used flow cytometric analysis of pooled chronic lymphatic leukaemia (CLL) cells to detect antibodies directed against HLA antigens. We show that FACS screening of pooled cells can accurately and rapidly detect these antibodies and that the method is suitable for routine use. An estimate of the degree of patient panel reactivity can be determined within a few hours. In addition, the technique is more sensitive than those conventionally used, an advantage that may be of importance in preventing graft damage.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2277
    Keywords: Flow cytometry, alloantibodies, kidney transplantation ; Alloantibodies, kidney transplantation, flow cytometry ; Kidney transplantation, alloantibodies, flow cytometry ; Screening, alloantibodies, kidney transplantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Flow cytometric screening of sera using pooled chronic lymphocytic leukaemia (CLL) cells has previously been reported as a quick method for detecting HLA antibodies of the IgG class. In this study we investigated the sensitivity of this method in the detection of IgG and IgM alloantibodies, and its performance in serum screening when compared to conventional microlymphocytotoxic screening. Results indicate that flow cytometric screening is more sensitive in the measurement of IgG alloantibodies by up to five doubling dilutions, whereas the converse is true for IgM. IgM autoantibodies were found not to be detectable by flow cytometry. By testing a large number of sera by both methods in parallel, we have found that a significant proportion of sera exhibiting no activity or IgM activity alone on cytotoxic screening contain IgG antibodies detectable with a pool of CLL cells on the flow cytometer.
    Type of Medium: Electronic Resource
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