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Biomedical subjects

M Eulitz

Publications and source records attributed to M Eulitz.

At least 55 records · Page 3Linked to original sources
PubMed1986

Amyloid kidney stones of uremic patients consist of beta 2-microglobulin fragments.

Urinary stones with amyloid structure, obtained from uremic patients, were analyzed according to molecular weight, amino acid sequence, and antigenic content. A major protein of approximately 7 kD, designated AB protein, was isolated by size exclusion using HPLC in 60% formic acid. AB protein reacted in immunodiffusion only with an antiserum to beta 2-microglobulin, with beta 2m spurring over AB protein. N-terminal amino acid sequence analysis defined two fragments homologous to beta 2m. One fragment commenced with Ile at position 7 and the other with Ser at position 20, with a cleavage point subsequent to a lysyl residue in both. It is concluded that beta 2m is a precursor of urinary amyloid stones and intratubular concretions of patients with preterminal and terminal renal failure; limited proteolysis is involved in AB amyloid generation.

Amino Acid Sequence↗
PubMed1986

The primary structure of bdellin B-3 from the leech Hirudo medicinalis. Bdellin B-3 is a compact proteinase inhibitor of a "non-classical" Kazal type. It is present in the leech in a high molecular mass form.

A proteinase inhibitor was isolated from extracts of the leech Hirudo medicinalis by gel filtration and anion exchange chromatography. This inhibitor is similar to the bdellins in that it blocks the activity of trypsin, plasmin and sperm acrosin but has a molecular mass, as estimated by SDS polyacrylamide electrophoresis, of about 20 kDa, whereas the bdellins have molecular masses in the range 5-6 kDa. It is therefore designated as high-molecular mass bdellin B-3 (HMB). The amino-acid sequence of the inhibitor was elucidated as far as position 56. This revealed that the molecule consists of a bdellin B-3 moiety, corresponding to the N-terminal 46 residues, which is then extended at the C-terminus by a polypeptide chain of the composition Asx15, Glx25, Gly6, Val, His26-27 and Lys4. It has been formerly concluded from a partial amino-acid sequence that bdellin B-3 is a Kazal-type inhibitor. However, the complete sequence of bdellin B-3, represented by the N-terminal 46 residues of HMB, discloses that bdellin B-3 is a non-classical Kazal-type inhibitor when the number of amino-acid residues between half-cystines are considered. Presuming that formation of disulfide bridges principally follows the same pattern as in classical Kazal-type inhibitors the bdellin B-3 molecule was modeled based on the known three-dimensional structure of the third ovomucoid domains. This showed that a compact arrangement of the peptide chain of bdellin B-3 is conceivable.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1985

Amyloid fibrils derived from V-region together with C-region fragments from a lambda II-immunoglobulin light chain (HAR).

Amyloid fibril proteins were isolated from the spleen of a patient with IgD(lambda)-plasmocytoma by extraction and gel filtration in 5M guanidine hydrochloride. The molecular mass of the predominant polypeptide chain was approximately 5000 Da. Its complete amino-acid sequence was elucidated by stepwise automated degradation of the carboxymethylated polypeptide chain and by structural studies of tryptic and thermolysinolytic cleavage products. The length of the polypeptide chain was 58 to 59 residues and it was homologous to the amino acids in positions 8 through 65 of the variable part of an lambda-type immunoglobulin light chain, which was most closely related to the lambda II subgroup. The N-terminal sequence of this amyloid fibril protein proved to be heterogeneous, indicating cleavage after the amino acids in positions 7 and 8. Peptides from the constant part of the lambda-chain were unexpectedly found in the tryptic digest of the denatured amyloid protein HAR. One polypeptide derived from the constant region was separated from the main component by high performance liquid chromatography. Its amino-acid sequence commenced at position 111 and could be traced in 41 steps. In this case, at least two constant region fragments were shown to be constituents of the amyloid fibril protein. The association of fragments from the variable as well as the constant region is discussed with respect to amyloid formation.

Amino Acid Sequence↗
PubMed1983

Antilymphocytic antibodies and marrow transplantation. VI. Absence of immunosuppression in vivo after injection of monoclonal antibodies blocking graft-versus-host reactions and humoral antibody formation in vitro.

The in vivo and in vitro effectiveness of several monoclonal antimouse T and B cell antibodies, of anti-Th-1 and of Iak serum, as well as of ATG were compared. The parameters were prolongation of skin graft survival, prevention of graft-versus-host disease (GVHD), antibody and primary and secondary plaque formation against sheep redblood cells (RBCs), and T cell depletion of lymphoid tissues. In general, in vitro effectiveness of the monoclonal antibodies exceeded their in vivo effectiveness. Skin graft survival was prolonged by ATG, but not by monoclonal anti-T, or anti-T plus anti-B antibody. GVHD was prevented by in vitro incubation of donor bone marrow with monoclonal anti-Th-1, but in vivo treatment of marrow donors was ineffective. Treatment with ATG was successful. Anti Iak antibody blocked plaque formation by spleen cells incubated with sheep RBCs, but had no effect on secondary plaque formation when given in vivo. Neither was there any in vivo effect of anti-Iak or anti-Th-1 on antisheep RBC agglutinin formation. ATG was effective in both of these assays, although its cytotoxic and complement-fixing titer did not exceed that of anti-Th-1 or anti-Iak. Although anti-Th-1 was cleared more rapidly from the serum of mice expressing the corresponding Th-1 alloantigen, than from mice with the noncorresponding alloantigen and although anti-Th-1 was shown to bind to the T cell areas of the lymphoid tissue, it did not--unlike ATG--deplete these areas of T cells. Possible reasons for the difference in effectiveness of in vitro and in vivo application of these monoclonal antibodies are discussed.

Animals↗
PubMed1983

Identification of amyloid A protein in a sporadic Muckle-Wells syndrome. N-terminal amino acid sequence after isolation from formalin-fixed tissue.

The amyloid of a patient (WAL) with a sporadic Muckle-Wells syndrome was analyzed in tissue sections and after it had been isolated from formalin-fixed tissue. The predominant amyloid fibril protein was the amyloid A (AA) type determined by the following criteria. (a) Only antiserum against protein AA gave a strong specific reaction, whereas the other antisera with specificity against amyloid of immunoglobulin origin, i.e., A-lambda and A-kappa, did not stain using the indirect immunoperoxidase technique on formalin-fixed, paraffin-embedded tissue sections. (b) In immunodiffusion, the predominant amyloid fibril protein from WAL isolated in pure form from formalin-fixed tissues precipitated in a line of identity with anti-AA and a purified and chemically identified protein AA from another patient. (c) Amyloid fibril protein from WAL had a molecular weight of 8000 to 9000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis and, thus, is in the same range as the commonly found protein AA. (d) The N-terminal amino acid sequence analysis was that of protein AA. In addition to the pure amyloid fibril protein AA (WAL), proteins of higher molecular weights with AA-antigenic determinants were also isolated. These proteins may represent protein AA in a complex form or protein AA linked to other proteins. Since an inflammation is the most likely cause of amyloidosis complicating the Muckle-Wells syndrome an antiinflammatory therapy (as in other AA-type amyloidoses) is recommended.

Adult↗
PubMed1982

Monoclonal antibodies against T-cell antigens studied by immunohistochemistry.

Fifteen monoclonal antibodies against different T-cell antigens were studied by immunohistochemistry in thymus, fetal thymus, fetal liver, palatine tonsils, and a few T-cell lymphomas. OKT 9 was identified as reacting with hemopoietic stem or precursor cells in fetal liver as well as with early B-determined lymphocytes in tonsillar germinal centres. OKT 10 labelled lymphocytes in thymus and surprisingly also the cytoplasm of some tonsillar cells with plasma-cell like appearance. OKT 6 and MAS 036 b reacted only with thymic cells. OKT 4, OKT 5, OKT 8,8-11, labelled thymic cells- and portions of interfollicular cells in tonsils. OKT 3, NEI 016, NEI 015, and T 28 stained a majority of thymic cells and of tonsillar interfollicular lymphocytes. IFH-M 203, NEI 012 and 4-11 were positive with the majority of T-lymphocytes in tonsils but labelled only a few thymic cells.

Animals↗
PubMed1982

Primary structure of the variable part of an amyloidogenic Bence-Jones Protein (Mev.). An unusual insertion in the third hypervariable region of a human kappa-immunoglobulin light chain.

The complete amino acid sequence of the variable region of Bence-Jones protein Mev. from a patient suffering from multiple myeloma and generalized amyloidosis is presented. The amino acid sequence of the Bence-Jones protein Mev. is related to other human kappa-immunoglobulin L-chains of subgroup I. With valine established in Position 191 of the constant region, it is of the Inv (3) allotype. Two types of the Bence-Jones protein Mev. were found, one beginning with the typical N-terminal aspartic acid, and another lacking the N-terminal tripeptide and commencing with methionine in Position 4. A unique insertion of glutamic acid after Position 95 was found in the Bence-Jones protein. This is the position where the V- and J-gene segments join. The J-region of Bence-Jones protein Mev. exhibits some marked differences to the five J-regions recently established by nucleic acid sequencing. This suggests, that there must be considerable polymorphism in human kappa-J-genes. The amyloid fibril protein from the same patient (A Mev.) has also been sequenced up to Position 27. It was found to be identical to the sequence of Bence-Jones protein Mev. commencing with aspartic acid. The molecular mass of the amyloid fibril protein was found to be between 11 000 and 12 000 Da as estimated by gelfiltration and dodecyl sulfate-polyacrylamide electrophoresis.

Amino Acid Sequence↗
PubMed1982

Complete amino acid sequence of the ADP/ATP carrier from beef heart mitochondria.

The complete primary structure of the ADP/ATP carrier from beef heart mitochondria is described. Cyanogen bromide cleaves the protein into a long, N-terminally blocked fragment with Mr 22,000 (CB1) and several small peptides. The primary information was derived from liquid-phase sequencing of tryptic peptides obtained from the maleylated carrier protein and the citraconylated CB1 fragment, as well as from cleavage products with Staphylococcus aureus protease. The multitude of thermolysinolytic, tryptic, chymotryptic and peptic peptides was sequenced by manual methods. They rendered overlaps and further supported already known partial sequences. Also, the bridge between the published acidolytic C-terminal fragment A2 was thus obtained.

Amino Acid Sequence↗
PubMed1981

Tritium radiolabeling of antibodies to high specific activity with N-succinimidyl [2,3-3H] propionate: use in detecting monoclonal antibodies.

A simple method has been established for tritium labeling immunoglobulins. This label avoids certain disadvantages of 125iodine such as high radiation hazard and requirement for frequent labeling owing to the short half-life of this isotope. High specific activities (range 1.8-9.0 cpm/mu g protein) were obtained with no loss of functional activity as demonstrated by passive hemagglutination. The labeled conjugate is especially suitable for detection of activity in hybridoma supernatants by indirect binding assay. Adapted to microtiter plates, the method is suitable for large numbers of samples, e.g. up to 800 supernatants may be examined daily. The utility and advantages of the binding assay with respect to reproducibility, sensitivity, binding kinetics, easy and rapid performance are described.

Animals↗
PubMed1980

Idiopathic amyloidosis in the stone marten (Martes foina): identification of amyloid fibril proteins in tissue sections using the immunoperoxidase technique.

Amyloid fibril proteins isolated from a spleen of a wild stone marten (Martes foina, Exleben) with idiopathic amyloidosis show resemblance to protein AA by amino acid analysis. An antiserum directed against these proteins can be used to identify the marten's amyloid in formalin-fixed tissue paraffin-embedded sections using the immunoperoxidase method.

Amyloid↗
PubMed1980

Structure of the elastase-cathepsin G inhibitor of the leech Hirudo medicinalis.

The leech Hirudo medicinalis contains three different groups of proteinase inhibitor proteins, the thrombin-specific hirudin, the bdellins directed against trypsin, plasmin and acrosin, and the eglins which were discovered only recently. We are interested in the eglins mainly for two reasons: (i) They form strong complexes with the granulocytic elastase and cathepsin G with Ki values close to 1 x 10(-10) mol/l. Due to this property they are potential candidates for the therapeutic treatment of various diseases. (ii) Although the eglins do not contain a disulfide bridge to stabilize the tertiary structure, they are highly resistant to denaturation by acidification and by heat as well as to proteolytic degradation.

Amino Acid Sequence↗
PubMed1979

[The primary structure of the Bence-Jones protein Kue. The amino acid sequence of the variable part of a human L-chain of the kappa-type (author's transl)].

The complete primary structure of the variable region of Bence-Jones protein Kue was elucitated with the aid of a few tryptic peptides, as well as one chymotryptic and one BNPS-scatol fragment. As a consequence of the evident homologies to other proteins this protein belongs to subgroup k/I. Protein Kue has some amino acid exchanges in certain positions in common with other proteins, probably giving rise to a new sub-subgroup. The constant region shows no amino acid exchanges in comparison with other human kappa L-chains. With valine covering position 191, protein Kue should be grouped per definition as an allotype Km (3).

Amino Acid Sequence↗
PubMed1978

Amino-acid sequence of toxin I from Anemonia sulcata.

Toxin I from Anemonia sulcata, a major component of the sea anemone venom, consists of 46 amino acid residues which are linked by three disulfide bridges. The [14C]carboxymethylated polypeptide was sequenced to position 29 by automated Edman degradation. The remaining sequence was determined from cyanogen bromide peptides and from tryptic peptides of the citraconylated [14C]carboxymethylated toxin. Toxin I is homologous to toxin II from Anemonia sulcata and to anthopleurin A, a toxin from the sea anemone Anthopleura xanthogrammica. These toxins constitute a new class of polypeptide toxins. No significant homologies exist with toxin III from Anemonia sulcata nor with known sequences of neurotoxins or cardiotoxins of various origin.

Amino Acid Sequence↗
PubMed1978

Demonstration of the Fc-receptor of blood cells by soluble peroxidase-anti-peroxidase (PAP) complexes.

The Fc-receptor of normal human leukocytes, of CLL-cells, and of hematopoietic cell lines was demonstrated with soluble peroxidase-anti-peroxidase (PAP) complexes. In about 9% of normal lymphocytes an almost continuous, strong labeling of the cell membrane was established. Some of these lymphocytes were characterized by a peculiar uniform fine structure. The percentage of PAP-labeled monocytes was in the range of 25%, neutrophils nearly 100%, eosinophils 0%, CLL-cells 10%. Labeled portions of the membrane were interiorized from monocytes. The lymphoid cell-line Daudi established from a Burkitt's lymphoma appeared almost negative, the cell line K562 established from a myeloid leukemia in 75% of the cells strongly positive. PAP-labeling was not influenced by preincubation with trypsine or with neuraminidase; it was negative when PAP-F(ab)2 was used. Results of PAP-labeling were not always in agreement with EA-rosettes or with agg-Ig.

Binding Sites↗
PubMed1978

Improved preparation and structural elucidation of the tryptophan cleavage reagent 2-(2'-nitro-phenylsulfenyl)-3-methyl-3-bromoindolenine (BNPS-skatole).

Methods are described for a high yield preparation (overall yield greater than 90%) of analytically pure 2-(2'-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine (BNPS-skatole). The chromatographic and spectroscopic data are given. The yield of tryptophan cleavage is considerably dependent on the degree of purity of the BNPS-skatole.

Chemical Phenomena↗
PubMed1976

Immunohistochemical identification of T- and B-lymphocytes delineated by the unlabeled antibody enzyme method. I. Anatomical distribution of theta-positive and Ig-positive cells in lymphoid organs of mice.

The unlabeled antibody enzyme method was used to delineate the anatomical distribution of lymphocytes positive with a MBtheta and a-MIg in tissue sections of thymus, spleen, lymph nodes and Peyer's patches of mice. The known T- and B-cell areas were established. Moreover, individual B-cells are detected in T-cell regions, especially in the thymus medulla, and in the periarteriolar section of spleen white pulp. Similarly, individual T-cells occur in B-cell areas, namely in the marginal zone of spleen white pulp, in the medullar cores of lymph nodes, and in germinal centers.

Animals↗
PubMed1975

[Quantitative 125-I-autoradiography of individual cells (author's transl)].

Iodine 125, an emitter of beta-radiation with an energy lying between that of tritium and carbon-14, is investigated for its applicability in quantitative autoradiography. Absorption and geometric factors of radiation are elucidated. From this, appropriate measuring conditions are derived. The simultaneous exposure of radioactive standard sources permits the evaluation of absolute amounts of radioactivity. Standard cells with labelled membranes are a suitable source of reference taking into account the physical properties of the isotope. Sheep red blood cells are examined for their suitability as standard cells after enzymatic radioiodination. The absolute number of antigenic substances on the surface of single cells is obtained by determining the specific activity of the labelled antibody molecules, and by measuring the silver grain densities of the cells under investigation and of the standard cells. The radioactivity per standard cell can be assessed by conventional procedures. The new method is applied to the quantification of membrane-bound immunoglobulin molecules of the IgG-type on single human lymphocytes. The determination of an immunologic saturation of the labelled antibody is essential for this purpose. On the lymphocytes of a normal person and of a patient with chronic lymphatic leukaemia quite different amounts of immunoglobulins have been evaluated.

Absorption↗