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Biomedical subjects

L B Sandberg

Publications and source records attributed to L B Sandberg.

At least 37 records · Page 2Linked to original sources
PubMed1989

Uniqueness of dietary olive oil in stimulating aortic prostacyclin production in post-weanling rats.

Groups of weanling Sprague-Dawley rats developed from conception through gestation, and weanling periods on a formulated diet fed to the dams were continued on the same diet until sacrificed at 30 days of age. The diet groups consisted of control (5% corn oil, w/w) and experimental (15%, w/w) olive, safflower (hi-oleic and hi-linoleic), soy oil, and lard. The object of the study was to identify the effect of high and low fat content and differing proportions of polyunsaturated:saturated (P:S) and mono:polyunsaturated (M:P) fatty acids on arachidonate stimulated aortic prostacyclin (PGI2) production (measured as 6-keto-PGF1 alpha). Neither the amounts of dietary fat or wide ranging P:S or M:P fatty acid ratio levels (P:S or M:P) affected PGI2 production. PGI2 production was, however, markedly enhanced (2x) in aortic segments from rats raised on diets containing olive oil. The unique stimulation of aortic PGI2 production by the olive oil diet suggests an effect of the extraordinarily high M:P fatty acid ratio or, alternatively, of a still-to-be identified substance(s) in this ancient food.

6-Ketoprostaglandin F1 alpha↗
PubMed1986

Quantitation of elastin through measurement of its pentapeptide content.

Digestion of insoluble porcine elastin with thermolysin produces a number of discrete small peptides. That present in highest concentration is the pentapeptide valyl-glycyl-valyl-prolyl-glycine (VGVPG) derived from the portion of the polymer containing extensive repeats of this sequence. Among eukaryotes, this sequence appears to be found only in elastin and its precursor tropoelastin. In the pig this is represented by peptide W4 of a tropoelastin tryptic digest (Sandberg, L.B., et al. Path. Biol. 33, 266-274, 1985). Quantitation of this peptide by HPLC separation, monitoring its absorption at 212 nm, offers a simple reliable means of measuring purified insoluble elastin as well as non-purified elastin in fat-free tissue samples. Digestion times and incubation temperatures are discussed. The method is sensitive enough to accurately quantitate elastin at the 2 to 3 microgram level.

Amino Acid Sequence↗
PubMed1986

Classification of malnutrition by statistical analysis of quantitative two-dimensional gel electrophoresis of plasma proteins.

An attempt to use the relative concentrations of major plasma proteins for clinical assessment of severe malnutrition is described. Quantitative two-dimensional gel electrophoresis was used to measure the concentrations of 24 major proteins in small aliquots of plasma obtained from children, aged 0 to 3 years, who were patients and outpatients in Liberian hospitals. Fifteen had a clinical diagnosis of kwashiorkor, 36 were diagnosed with marasmus, and 18 were controls. There were also 5 controls from the United States. The individuals were placed in six groups; kwashiorkor, kwashiorkor who died during treatment, marasmus, marasmus who died, Liberian controls, and U.S. controls. The amount of protein in each spot in the two-dimensional gels was estimated by measuring bound stain using a laser scanner and computerized image analysis. We found very low serum transferrin levels in malnourishment, in agreement with reports from other investigators. All of the data for 24 protein variables were pooled for factor analysis; the mean factor scores for each group differed, with the kwashiorkor groups furthest from the controls. Results of discriminant analysis using the amounts of different numbers of protein variables (3 to 24) were compared for posterior assignment of individuals to groups. The validity of the method was tested by analysis of plasma aliquots obtained from patients following initiation of therapy and which were not a part of the training set. Predictive performance (prognosis of patient survival) depended upon the number of protein variables used. Although artifactual fitting of the data is expected to contribute to performance as the number of variables is increased, use of as many as 7 variables may be justified, even with our small patient groups. Possible use of these results for development of a practical clinical test is discussed.

Blood Protein Electrophoresis↗
PubMed1985

Lysyl oxidase activity in lungs of copper-deficient hamsters.

Lysyl oxidase, the enzyme responsible for mediating crosslink formation in collagen and elastin, requires copper for its activity. In this study, lysyl oxidase activity and insoluble elastin content were unchanged in lungs from copper-deficient hamsters compared to controls. The lack of dramatic diminution in lysyl oxidase activity in animals who demonstrate significant structural alterations in the lung suggests that other mechanisms in addition to inhibition of crosslink formation are operative in this model.

Amino Acid Oxidoreductases↗
PubMed1985

Elastin covalent structure as determined by solid phase amino acid sequencing.

The amino acid sequences of 16 large tryptic fragments of aortic tropoelastin have been determined establishing the presence of several repeating structures: GVP, GGVP, PGVGV, PGVGVA, and AGVPGFGVG. The methodologies for achieving these results by solid phase sequencing are reviewed and also the possible biologic significance of the unusual primary structures of elastin are discussed.

Amino Acid Sequence↗
PubMed1984

Quantitation of elastin production in cultured vascular smooth muscle cells by a sensitive and specific enzyme-linked immunoassay.

An enzyme-linked immunoassay (ELISA) procedure has been developed to quantitate the amount of elastin produced by cultured porcine aortic smooth muscle cells. ELISA was used to determine both titer and specificity of antisera raised in rabbits against porcine aortic alpha-elastin conjugated with key-hole limpet hemocyanin. Under optimum conditions (1: 3000 dilution of antiserum, 20 ng alpha-elastin per assay well), sensitivity averaged 60 ng/ml). Specificity was confirmed by immunoprecipitation of [125I]-tropoelastin, radial immunodiffusion, Western blotting and lack of cross-reactivity with serum proteins or collagen. Extensive cross-reactivity was found with both human alpha-elastin and porcine beta-elastin, while porcine tropoelastin was able to compete with 80% of the alpha-elastin determinants. Affinity of anti-porcine antisera for sheep alpha- elastin was significantly lower. When the ELISA was made specific for tropoelastin by coating wells with 60 ng of this antigen, a time-dependent and serum-dependent rate of production of tropoelastin was observed in the culture medium of primary and secondary cultures of smooth muscle cells. Comparison of elastin production in cultures of porcine smooth muscle cells suggests that porcine aortic elastin production varies as a function of cell density and phase of growth.

Animals↗
PubMed1983

The influences of sample distribution and age on reference intervals for adult males.

This investigation was designed to improve reference information and to evaluate the influences of sample distribution and age on the derived reference intervals. Specimens from 127 men were collected after a 12- to 14-hour fast and analyzed by 60 different laboratory procedures. Differences in the reference intervals derived, using three separate statistical methods, appeared to be unimportant clinically, but the percentile method was preferred because it required no assumptions concerning the frequency distribution. Relationships between age and analyte concentrations were examined by linear regression analysis, and the analytes were placed in one of three groups, according to the significance of this relationship: greatest significance (P less than or equal to 0.01), 18 analytes; intermediate significance (0.01 less than or equal to P less than or equal to 0.05), 12 analytes; and least significance (P greater than 0.05), 30 analytes. The age-related changes for each analyte were evaluated according to analytic variation, population dispersion, and clinical relevance. Age-dependent reference intervals for adult males are recommended for albumin, cholesterol, phosphorus, and sedimentation rate.

Adult↗
PubMed1982

Structure of the elastic fiber: an overview.

Intense research efforts over the past 18 yr have probed deeply into the structure of the elastic fiber. This began with the elucidation of the demosine crosslinks in elastin and the description of the elastin precursor, tropoelastin, derived from copper-deficient animals. Characterization of the precursor material indicates that it is a single polypeptide chain of approximately 800 amino acid residues containing lysine residues in clusters destined to form the desmosine crosslinks. The molecule contains large areas of hydrophobic sequence interspersed with shorter stretches of polyalanine and the lysines. The shorter structures may be folded into alpha-helices. The larger hydrophobic areas appear to form a unique structure known as the beta spiral which possesses elastometric properties. Inside the hydrophobic areas repeating sequences such as the pentapeptide pro-gly-val-gly-val have been observed the exact significance of which is not appreciated, but it appears to be well-conserved between species. Recent studies in the molecular biology of this protein have indicated that it is synthesized on the rough ER with a short leader sequence of about 25 residues. This is lost before the tropoelastin is exported. Diversity in sequence studies in these leaders suggest that there may be two elastins, type A and B, which vary with the maturation of the animal.

Amino Acid Sequence↗
PubMed1982

A copper-deficient, zinc-supplemented diet produces emphysema in pigs.

A mild form of emphysema was produced in pigs raised on a copper-deficient, zinc-supplemented diet. The copper-requiring enzyme, lysyl oxidase, catalyzes the cross-linking of tropoelastin into mature elastin. Zinc further inhibits the activity of lysyl oxidase. Lungs from animals raised on copper-deficient, zinc-supplemented diets of demonstrate perforations in alveolar walls and diminished amounts of elastin bronchi and pulmonary arteries. Mean linear intercepts are greater and alveolar internal surface areas are less than those in control animals, fulfilling the generally accepted definition of emphysema. Physiologic confirmation is provided by a leftward shift of the saline volume-pressure curves when compared with those in control animals. Ultrastructurally, the alveolar walls are effaced and pores of Kohn are enlarged. There are areas in which elastin is absent leaving remnant microfibrils, and there are other changes consistent with active elastin synthesis. Biochemical data demonstrate no difference in elastin content as micrograms/ml of fat-free dry weight but do demonstrate increased collagen content in experimental animal lungs compared with that in control lungs. Ultrastructural similarities to enzyme-induced models of emphysema suggest the presence of elastin degradation in our model. We speculate that although the copper-deficient, zinc-supplemented state may stimulate protein synthesis in general, elastin is being degraded by endogenous means, but collagen is not.

Air Pressure↗
PubMed1982

The synthesis of elastin, collagen, and glycosaminoglycans by high density primary cultures of neonatal rat aortic smooth muscle. An ultrastructural and biochemical study.

Primary cultures of neonatal rat aortic smooth muscle cells inoculated at high densities (1 X 10(6) cells/25 cm2 Falcon flask) with adequate nutrient media and pH control grow rapidly and form multilayers of cells with typical "hill and valley" organization. After 10 days growth insoluble elastin formation could be visualized by phase contrast microscopy as small particles which grew rapidly to become larger irregular refractile aggregates and later coalesced to form larger aggregates and small fibres. With light and electronmicroscopy, elastin was the predominant matrix protein formed, with the "hill regions" of cultures containing abundant elastin aggregates and some collagen. In 2-week-old cultures differentiation could be observed within the cell multilayer. The older deeper cells contained more protein synthesis organelles and myofilaments and were in close association with large often coalescing elastin aggregates; compared to younger more superficial cells which contained more free polyribosomes less myofilaments, and were associated with fewer and small elastin aggregates. In older cultures this differentiation was not apparent; the cells contained many myofilaments, dense bodies, and lysosomes. Elastin aggregates and newly formed elastic fibres were abundant in the matrix. Quantitative analysis of insoluble elastin formation in the cell layer during the 4-week culture period indicated continuous biosynthesis and deposition which paralleled that of desmosine formation. Amino-acid analysis of a hot alkali insoluble residue (regarded as elastin) from 30-day-old cultures gave a profile identical with neonatal rat aortic elastin in vivo. Insoluble collagen formation in the cell layer tended to plateau after the log phase of growth was completed (10 days). Proteoglycans were found predominantly in the supernatant media. Glycosaminoglycan analysis revealed a profile of dermatan sulphate (32%), chondroitin 4-sulphate (43%), keratan and heparan sulphate (30%), with only a trace of hyaluronic acid. This study indicates that primary cultures of neonatal rat aortic smooth muscle cells remain differentiated in culture and have the unique capacity to continue to synthesize and deposit large amounts (mg) of insoluble elastin which aggregate and from elastic fibres in vitro.

Animals↗
PubMed1981

In vitro studies of elastin metabolism.

Neonatal pig and rat aortas were studied for their ability to synthesize elastin in an in vitro situation. Smooth muscle cells from the rat aorta produced excellent multilayered cultures and produced soluble elastin (tropoelastin), insoluble elastin, and small amounts of collagen. BAPN proved to be toxic to these cells, adversely affecting the level of extracellular protein production. Tissue minces from pig aorta continued to synthesize elastin for two hours after removal. However, a 24 hour study indicated that elastin synthesis had almost completely shut down and that collagen synthesis continued in an apparently normal fashion. It is concluded that in vitro elastin synthesis is an extremely sensitive process easily altered by culture conditions and the addition of extraneous substances such as BAPN, and also highly influenced by the past history of the smooth muscle cells involved.

Animals↗