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  • 1
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 41 (1995), S. 0 
    ISSN: 1365-3083
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Bovine serum amyloid-A (SAA) was purified from acute-phase high density iipoprotein (HDL) by affinity chromatography and subsequent gel filtration chromatography. The identity of the isolated protein was checked by Western blotting following SDS-urea-PAGE using antisera raised against the purified protein fraction (SAA) and Amyloid A (AA). The antiserum raised against the purified SAA stained Congo red positive regions in the kidney of an AA-amyloidotic cow and reacted on Western blot with an AA-related protein of approximately 14kDa. Moreover, it immunostained two to three bands, of approximately 14kDa, present in serum from diseased cows, proportionally to the serum SAA concentration as measured by ELISA.Isoelectric focusing of the purified bovine SAA fraction revealed three major (pI 5.5, 6.0, 6.4) and three minor (pi 4.8, 5.0, 7.3) isoforms and two-dimensional SDS-urea-PAGE confirmed the identity of the major isoforms. Isoelectric focusing of SAA isolated from sera, obtained from cows affected with different diseases, showed a variable ratio of the isoforms. In SAA isolated from serum obtained from acow suffering from spontaneous AA-amyloidosis only one isoform (pI 4.8) was detectable.It is concluded that the results give first evidence for the existence of multiple isoforms of bovine SAA, occurring in different plasma concentration ratios during different diseases.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 37 (1993), S. 0 
    ISSN: 1365-3083
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Tumour necrosis factor-α (TNF-α) is one of the cytokines that stimulate the production of serum amyloid A (SAA), the precursor of AA amyloid. The role of TNF-α in amyloidogenesis was investigated in experimental hamsters using purified recombinant human TNF-α (rhTNF-α) and rhTNF-α analogue different from the normal molecule by two amino acid substitutions. Daily injections of 1 μg rhTNF-α resulted in elevated SAA levels but even in the presence of amyloid enhancing factor (AEF) no amyloid was deposited, indicating that apart from the AEF and one particular SAA stimulating factor an additional factor is needed to result in amyloid deposition. This factor is generated by repeated injections of E. coli lipopolysaccharide (LPS).A single intraperitoneal injection of 12,5 μg or more of rhTNF-ä followed by seven daily subcutaneous injections of LPS resulted in enhanced amyloid deposition. Heat denaturation of rhTNF-α did abolish its AEF activity. The rhTNF-α analogue, having one-fifth of the cytotoxic activity of the normal rhTNF-α, showed a similar reduction in its SAA-inducing capacity and its amyloidogenicity. This suggests the AEF activity to be closely related to TNF-α activity. However, poly(I)poly(C) (a potent inducer of IL-6) also showed AEF activity, suggesting that not a single cytokine but rather a certain combination of different cytokines could be decisive in AA amyloidogenesis.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    ISSN: 1365-3083
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Cells of the mononuclear phagocytic system (MPS) were described as playing a decisive role in amyloidogenesis. A relationship between the amyloid enhancing factor (AEF) and MPS cells was suggested and recently AEF activity was attributed to a serine esterase (SE) of leucocytic origin. In the present study, no correlation was found between the SE content and AEF activity in either peritoneal cell lysates or AEF preparations of different origin. Furthermore, pretreatment of fibril AEF (FAEF) with the SE inhibitor phenylmethylsulphonyl fluoride (PMSF) did not affect its activity in the hamster. Blockade of the MPS by dextran sulphate did not inhibit deposition of amyloid after intravenous injection of FAEF but amyloid deposition was inhibited when FAEF was administered intraperitoneally. These results suggest that MPS cells could be involved in transport of AEF, but that phagocytic activity of MPS cells is not essential in AA-amyloid fibrillogenesis. It is concluded that these results are not consistent with the previously suggested nature of the AEF or with the proposed central role of the MPS in amyloidogenesis.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 39 (1994), S. 0 
    ISSN: 1365-3083
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: The nearly complete amino acid sequence of casein-induced hepatic hamster AA is described. Hamster AA appeared to be C-terminally ragged as found in other species. Furthermore, the N-terminus was (partially) truncated. Hamster hepatic AA sequence showed high homology with, but was not identical to hamster SAA-isotype sequences deduced earlier from mRNA in hepatic and extra-hepatic tisstie. Therefore, hamster hepatic AA is derived from a novel member of the SAA gene family. However, the origin of hepatic AA-amyloid remains unclear.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 5
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 31 (1990), S. 0 
    ISSN: 1365-3083
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: High-density lipoprotein (HDL) apolipoprotein was separated from hamster serum by cholesteryl hemisuccinate atlinity chromatography (CHAC) in comparison with the density-gradient ultracentrifugation (DGUC), The apolipoprotein recovery from scrum by CHAC was 70% and by DGUC 80%, This disadvantage is compensated for by the ease of purification by CHAC, a method particularly suited for the processing of large amounts of serum.From the acute-phase HDL CHAC fraction, apo SAA was isolated by gel filtration. Using isoelectrofocusing, two-dimensional gel electrophoresis, und titration curve, four isotypes of hamster apo SAA were identified and characterized In the acute-phase serum, one of the isotypes was predominant (apo SAA1), In serum of amyloidotic animals, the relative contribution of apo SAAl was considerably lower, suggesting selective removal of the latter during amyloidogencsis and possibly its deposition in hamster AA amyloid. Furthermore, the affinity chromatography method was modified with gradient elution of affinity-bound material. By this method HDL apolipoprotein was separated into three subclasses. Apo SAA was shown to associate with two different subclasses. In acute-phase serum most of the apo SAA1 was found in the subclass with the lowest affinity for the cholesteryl beads, whereas thc latter was depressed in amyloidotic serum, suggesting that the amyloidogenicity ofa particular apo SAA isotype is determined by its cholesteryl-binding properties.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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