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Biomedical subjects

B R Olsen

Publications and source records attributed to B R Olsen.

At least 199 records · Page 11Linked to original sources
PubMed1980

Purification and characterization of the amino-terminal propeptide of Pro alpha 1(I) chains from embryonic chick tendon procollagen.

A peptide with an apparent molecular weight of 23,000 was isolated from the medium of cultured chick embryo tendons. Comparison of tryptic peptides derived from the medium peptide and from the amino-terminal, bacterial collagenase resistant portion of Type I procollagen indicated that the medium peptide represented the amino-terminal precursor-specific region of the pro alpha 1 (I) chain of procollagen. This conclusion was supported by the demonstration that antibodies against the medium peptide reacted with Type I procollagen in a radioimmune assay but did not react with a peptide derived from the carboxy-terminal propeptide of Type I procollagen. In addition, the reaction with Type I procollagen was inhibited with the purified amino-terminal, collagenase-resistant portion of pro alpha 1 (I) chains. Finally, amino acid sequencing demonstrated that the amino propeptide of dermatosparactic calf pN alpha 1 (I) chains and the medium peptide have similar amino-terminal sequences. Carbohydrate analysis established the presence of one residue of N-acetylglucosamine and a trace of mannose and galactose.

Amino Acid Sequence↗
PubMed1979

Butvar B-98 as a thin support film.

Support films prepared from butvar B-98 resin are mechanically stable, electron transparent, and possess minimum intrinsic structure. A simple procedure for routine preparation of support films using this resin is provided.

Microscopy, Electron↗
PubMed1978

Immunohistologic analysis of fetal and dermatosparactic calf and sheep skin with antisera to procollagen and collagen type I.

Immunofluorescent analysis of bovine and sheep skin using antibodies to the biosynthetic precursor pn-collagen type I, demonstrated that the localization of pn-collagen is restricted to the stratum papillare in adult skin. In contrast, antibodies against pn-collagen type I stained the whole dermis in skin obtained from dermatosparactic cattle and sheep. Electron microscopy of dermatosparactic sheep skin stained with ferritin-labeled anti-pn-collagen type I antibodies further demonstrated that pn-collagen molecules are the structural components of the unusual, twisted collagen fïbers observed in dermatosparactic skin.

Animals↗
PubMed1977

Purification and characterization of a peptide from the carboxy-terminal region of chick tendon procollagen type I.

A disulfide-bonded peptide with a molecular weight of about 100 000 was isolated from the medium of cultured chick embryo tendons. It was shown to be a trimer with two types of subunits in a 2:1 ratio, and tryptic fingerprinting and immunological evidence indicated that it was derived from the carboxy-terminal-precursor-specific region of procollagen. Amino acid analysis after reduction and alkylation indicated that the trimer contains about 30 residues of half-cystine involved in intrachain as well as interchain disulfide bonding. The interchain bonds could be reduced and alkylated under nondenaturing conditions. Carbohydrate analysis showed that each of the three peptide chains in the trimer contains about two residues of N-acetylglucosamine and about ten residues of mannose. This suggests the presence of one or two oligosaccharide units per chain.

Acetylglucosamine↗
PubMed1976

Characterization of the amino-terminal segment in type III procollagen.

Native type III collagen and procollagen were prepared from fetal bovine skin. Examination of the cleavage products produced by digestion with tadpole collagenase demonstrated that the three palpha1(III) chains of type III procollagen were linked together by disulfide bonds occurring at both the amino-terminal and carboxy-terminal portions of the molecule. Type III collagen contained interchain disulfide bonds only in the carboxy-terminal region of the molecule. After digestion of procollagen with bacterial collagenase an amino-terminal, triple-stranded peptide fragment was isolated. The reduced and alkylated chain constituents of this fragment had molecular weights of about 21 000. After digestion of procollagen with cyanogen bromide a related triple-stranded fragment was isolated. The chains of the cyanogen bromide fragment had a molecular weight of about 27 000. When the collagenase-derived peptide was fully reduced and alkylated, it became susceptible to further digestion with bacterial collagenase. This treatment released a fragment of about 97 amino acid residues which contained 12 cystein residues and had an amino acid composition typical for globular proteins. A second, non-helical fragment of about 48 amino acid residues contained three cysteines. This latter fragment is formed from sequences that overlap the amino-terminal region in the collagen alpha1(III) chain by 20 amino acids and possesses an antigenic determinant specific for the alpha1(III) chain. The collagenase-sensitive region exposed by reduction comprised about 33 amino acid residues. It was recovered as a mixture of small peptides. These results indicate that the amino-terminal region of type III procollagen has the same type of structure as the homologous region of type I procollagen. It consists of a globular, a collagen-like and a non-helical domain. Interchain disulfide bonding and the occurrence of cysteines in the non-helical domain are, however, unique for type III procollagen.

Amino Acid Sequence↗
PubMed1976

Medullary carcinoma of the thyroid. Diagnostic problems.

Light microscopy is usually considered sufficient for the diagnosis of medullary carcinoma of the thyroid (MCT). As stromal amyloid is not always present and the tumor may exhibit great variation in growth pattern, light microscopy alone, however, may lead to misinterpretations. Of 1670 thyroid carcinomas registered durging a 15-year period in Norway, 42 were originally interpreted as MCT. The slides were reviewed and the diagnosis maintained in 33 cases only. Twenty-four additional cases were found by reviewing histopathologic slides from neoplasms originally registered as other types of thyroid tumors. Of 57 cases of MCT, stromal amyloid was demonstrated in 43. Calcitonin measurements and electron microscopy, even on formalin-fixed material, were valuable aids in establishing the correct diagnosis, though none of these methods are unequivocal. Different aspects of the problems concerning the diagnosis of MCT are discussed through the detailed presentation of five patients.

Aged↗
PubMed1976

Segment-long-spacing aggregates and isolation of COOH-terminal peptides from type I procollagen.

Type I procollagen secreted by matrix-free cells from chick embryo tendons was purified by DEAE-cellulose chromatography. Electron microscopy of segment-long-spacing aggregates of the procollagen demonstrated the presence of both NH2-terminal and COOH-terminal extensions not found in collagen. The procollagen was digested with bacterial collagenase and the COOH-terminal fragments were isolated by gel filtration and polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Analysis of tryptic peptides demonstrated that the COOH-terminal extensions on the pro alpha 1 and pro alpha 2 chains had different primary structures.

Amino Acid Sequence↗
PubMed1976

Ultrastructure of parietal cells before and after proximal gastric vagotomy in duodenal ulcer patients.

In order to examine whether the reduced acid secretion observed after proximal gastric vagotomy (PGV) is accompanied by intracellular changes in parietal cells, the ultrastructure of parietal cells before and after PGV has been studied in four duodenal ulcer patients. A stereological analysis of electron micrographs shows that after PGV there is a reduction in the volume density of microvilli and the surface density of canalicular membranes. Ten days after PGV there is a temporary increase in the surface density of tubulovesicular membranes. However, 3 months after the operation most of this membrane material has been removed so that the surface density of tubulovesicles at this late stage is only insignificantly higher than before the operation. The relative volume of dense bodies is more than doubled 3 months after PGV, and the observed removal of tubulo-vesicular membranes between the 10 days and the 3 months stage may therefore be due to lysosomal activity.

Cell Membrane↗
PubMed1975

Two improved methods for preparing ferritin-protein conjugates for electron microscopy.

Two improved procedures were developed for activating ferritin so that the ferritin could be covalently linked to antibodies. One procedure involved use of a water-soluble carbodiimide and N-hydroxysuccinimide to prepare ferritin-containing activated esters. The other involved activation of the ferritin with excess glutaraldehyde. The ferritin-antibody conjugates prepared with the two procedures were shown to have a number of properties which made them suitable for locating antigenic components in cells.

Animals↗
PubMed1975

Further characterization of embryonic tendon fibroblasts and the use of immunoferritin techniques to study collagen biosynthesis.

Morphological studies were carried out on fibroblasts from chick embryo tendons, cells which have been used in a number of recent studies on collagen biosynthesis. The cells were relatively rich in endoplasmic reticulum and contained a well-developed Golgi complex comprised of small vesicles, stacked membranes, and large vacuoles. Techniques were then devised for preparing cell fragments which were penetrated by ferritin-antibody conjuates but which retained the essential morphological features of the cells. Finally, the new procedures were employed to develop further information as to how collagen is synthesized. As reported elsewhere, preliminary studies with ferritin-labeled antibodies showed that prolyl hydroxylase was found in the endoplasmic reticulum of freshly isolated fibroblasts and that procollagen is found in both the cisternae of the endoplasmic reticulum and the large Golgi vacuoles. In the experiments described here, the cells were manipulated so that amino acids continued to be incorporated into polypeptide chains but assembly of the molecule was not completed because hydroxylation of prolyl and lysyl residues was prevented. The results indicated that these manipulations produced no change in the distribution of prolyl hydroxylase. Examination of the cells with ferritin conjugated to antibodies which reacted with protocollagen, the unhydroxylated form of procollagen, demonstrated that protocollagen was retained in the cisternae of the endoplasmic reticulum during inhibition of the prolyl and lysyl hydroxylases. Assays for prolyl hydroxylase with an immunologic technique demonstrated that although the enzyme is found within the endoplasmic reticulum, it is not secreted along with procollagen. The observations provided further evidence for a special role for prolyl hydroxylase in the control of collagen biosynthesis.

Animals↗
PubMed1975

Location of procollagen in chick corneal and tendon fibroblasts with ferritin-conjugated antibodies.

Three distinct antiprocollagen preparations were characterized and used in immunocytochemical staining of chick embryo corneal and tendon cells. The several ferritin-conjugated antibody preparations permitted similar location of procollagen in the cisternae of the rough endoplasmic reticulum and in Golgi elements in both cell types. The ability to demonstrate and interpret specific ferritin staining was dependent on the extent of membrane breakage in each of those organelles, coupled with adequate retention of cell morphology. Corneal fibroblasts appeared to suffer more extensive intracellular membrane damage under controlled conditions of homogenization than tendon fibroblasts, facilitating the identification of procollagen in Golgi vacuoles of these cells. None of the labeled material appeared to by cytoplasmic in origin since ferritin was observed in the cytoplasm only in the vicinity of Golgi elements that were extensively broken. This study extends previous immunological evidence for the presence of procollagen in the Golgi complex and calls attention to the problems to be encountered in locating the antigen in small Golgi vesicles and lamellae.

Animals↗
PubMed1974

Ferritin-conjugated antibodies used for labeling of organelles involved in the cellular synthesis and transport of procollagen.

An improved procedure was used to conjugate ferritin to antibodies specific for the NH(2)-terminal extensions on the precursor form of collagen known as procollagen. The ferritin-antibody conjugates were then used to determine the localization of procollagen in fibroblasts isolated from chick-embryo tendons. Procollagen was found in the cisternae of the endoplasmic reticulum, indicating that the protein passes into this compartment early in its biosynthesis. Specific labeling of large Golgi vacuoles was also observed, suggesting that this compartment is also involved in the secretory process. When cells were incubated with colchicine so that the secretion of procollagen was delayed, there was an increase of large, smooth-surfaced vacuoles in the cells and these vacuoles were labeled with the ferritin-antibody conjugate.

Animals↗