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Biomedical subjects

M Adam

Publications and source records attributed to M Adam.

At least 145 records · Page 8Linked to original sources

Calcification of poly(2-hydroxyethyl methacrylate)-collagen composites implanted in rats.

Samples of the polyHEMA-collagen composites with varying collagen content have been implanted into the popliteal region of rats. Three, six and twelve months after the implantation, calcification of the implanted material was determined using a radioactive indicator. At the same time, the implants and surrounding tissue were examined histologically. The degree of calcification of the implants was dependent on the collagen content; it was more pronounced with a higher amount of collagen. The composites with 30% (w/w) or more collagen were biodegraded during the long-term implantation. It is suggested that the composites containing less than 20% (w/w) of fibrillar collagen are used for biomedical applications and that those with a higher collagen content for the in vitro studies.

Animals↗

The metabolism of [18F]6-fluoro-L-3,4-dihydroxyphenylalanine in the hooded rat.

The metabolism of the positron-emitting compound [18F]6-fluoro-L-3,4-dihydroxyphenylalanine (*F-DOPA) was studied in carbidopa-pretreated male hooded rats. Thirty minutes following carbidopa administration (5 mg/kg i.p.), animals received *F-DOPA (500 micrograms/kg; specific activity, 175-230 Ci/mol) as an intrajugular bolus. Blood samples were taken at various times between 5 and 90 min, and the plasma was analyzed by HPLC with gamma counting of fractions. *F-DOPA disappeared rapidly from plasma in concert with the formation of the 3-O-methylated metabolite, Me-*F-DOPA. Animals were killed from 5 to 120 min after injection, and the brains were rapidly dissected. The disappearance of *F-DOPA from both vermis and striatal samples was rapid. Me-*F-DOPA, the sole metabolite observed in the vermis, was the major labeled material in the striatum at greater than or equal to 20 min after injection. Fluorodopamine was an important metabolite in the striatum, making up 25% of total radioactivity at early intervals. Striatal samples also contained fluoro-3,4-dihydroxyphenylacetic acid, which constituted approximately 10% of the total radioactivity, and traces of two radiolabeled compounds, tentatively identified as fluorohomovanillic acid and fluoro-3-methoxytyramine.

3,4-Dihydroxyphenylacetic Acid↗

Externalization of membrane-bound activities during sheep reticulocyte maturation is temperature and ATP dependent.

During the maturation of sheep reticulocytes in vitro, there is release of material that can be pelleted from the cell-free incubation medium by centrifugation at 100,000 X g. This pellet contains activities that are derived from both the plasma membrane and lysosomes. No evidence was obtained for the presence of mitochondrial activities or cytosolic enzyme activities. The release of these activities is ATP and temperature dependent, since reduction of either results in a greater retention of the activities by the cells and a lesser amount in the 100,000 X g pellet. The pelleted material is vesicular in nature, and the production and (or) release of the material are reduced upon ATP depletion or lowering of the temperature. It is concluded that the externalization of specific membrane components is a normal metabolic process that occurs during reticulocyte maturation and represents a means by which reticulocytes shed specific types of membrane-associated functions that are known to decrease during reticulocytes maturation.

Adenosine Triphosphate↗

Effect of tolfenamic acid on the metabolism of the main connective tissue components in rats.

Tolfenamic acid (Clotam) has been used in the therapy of rheumatic diseases for some years. Regarding its chemical structure it belongs to the group of fenamates. The effect of tolfenamic acid on the synthesis of collagen and proteoglycans in granulation tissue, skin or cartilage of rat weanlings was tested and compared with the action of mefenamic acid. According to the results obtained, tolfenamic acid is a potent inhibitor of collagen as well as proteoglycan syntheses. The concentrations of the constituents of proteoglycans, i.e. protein core, link protein as well as glycosaminoglycans were decreased in the tissue after treatment with tolfenamic acid. In comparison with mefenamic acid, if the same doses were used (50 mg and 100 mg/kg of body weight/day in in vivo experiments and 10 mg/g wet tissue in in vitro experiments), tolfenamic acid exhibits more distinct inhibitory effect. A general inhibitory effect of tolfenamic acid on proteosynthesis is suggested.

Animals↗

Hepatitis-B as a prognostic discriminant in patients with primary liver cancer.

The prognostic significance of positive serologic determination of hepatitis-B was investigated in 137 patients with primary liver cancer seen over a 6-year period in Pretoria. The median survival time of the 137 patients was 93 days. Patients who were anicteric had a longer survival. Patients who showed positive results for hepatitis-B surface antigen (HBsAg) had a median survival time of 135 days, (39 patients) whereas patients who were HBsAg-negative had a median survival time of 96 days (66 patients). In 57 patients, one or more of the serologic determinations, HBsAg, HBeAg, anti-HBs, anti-HBe, and/or anti-HBc, were positive. Their median survival was 118 days. When HBsAg positivity (or negativity) is corrected for bilirubin, the median survival time remains a function of the bilirubin.

Adult↗

Evidence for a pool of non-recycling transferrin receptors in peripheral sheep reticulocytes.

Sheep reticulocytes from phlebotomized animals have a total transferrin binding potential that may exceed by an order of magnitude the surface binding capacity. Steady state uptake of transferrin at 37 degrees C is generally less than 50% of the total transferrin binding capacity. During long-term incubation of the reticulocytes, all transferrin binding ability is lost, the ability to internalize being lost most rapidly. The loss in ability to bind transferrin during long-term incubation is independent of the number of surface transferrin binding sites, since removal of surface receptors with pronase does not affect the rate of loss of the internal pool of receptors during long-term incubation. Moreover, after removing surface receptors with pronase, only a fraction of the original number of receptors is restored to the surface, despite the presence of a large pool of internal receptors. These data suggest that only a fraction of the internal pool of receptors is capable of recycling to the cell surface in sheep reticulocytes.

Animals↗

Monosaccharide composition of lipopolysaccharides from Campylobacter jejuni and Campylobacter coli.

The monosaccharide composition of the LPS from 5 Campylobacter jejuni strains and 7 Campylobacter coli strains has been studied. All LPS's contained KDO, heptose, glucosamine, glucose, and (with one exception) galactose. All C. jejuni and 3 C. coli LPS's contained greater than 1% galactosamine. 3-Amino-3.6-dideoxyglucose was present in all but one C. coli LPS and in only one C. jejuni LPS.

Campylobacter↗

Determination of larger urinary peptides in osteoarthrosis by high-performance liquid chromatography.

Urinary peptides larger than 4000 were fractionated by HPLC on reversed phase. Urines of osteoarthrotic patients were compared with those of healthy persons and rheumatoid patients. The profile of urinary peptides of patients with mild form of osteoarthrosis differed significantly from that of other groups studied. The results of patients with rheumatoid arthritis were similar to those of patients with activated or severe form of osteoarthrosis.

Adult↗

Poly(2-hydroxyethyl methacrylate)--collagen composites which promote muscle cell differentiation in vitro.

A new simple method has been developed which allows the mixing of poly(2-hydroxyethyl methacrylate)--polyHEMA--and fibrillar collagen in any desired ratio. PolyHEMA alone was shown to be an unsuitable cultivation substrate for primary cultures of chicken embryonic skeletal muscle cells. Composites containing polyHEMA and 50% (w/w) or more collagen supported myogenesis. Such layers, firmly adhered to the bottom of plastic Petri dishes, were mechanically stable and biologically active, thus favourably combining properties of both the original materials. It is suggested that polyHEMA-collagen composite layers may be used for cultivation of differentiating cells in vitro.

Animals↗

Electron microscopic evidence for externalization of the transferrin receptor in vesicular form in sheep reticulocytes.

Using ferritin-labeled protein A and colloidal gold-labeled anti-rabbit IgG, the fate of the sheep transferrin receptor has been followed microscopically during reticulocyte maturation in vitro. After a few minutes of incubation at 37 degrees C, the receptor is found on the cell surface or in simple vesicles of 100-200 nm, in which the receptor appears to line the limiting membrane of the vesicles. With time (60 min or longer), large multivesicular elements (MVEs) appear whose diameter may reach 1-1.5 micron. Inside these large MVEs are round bodies of approximately 50-nm diam that bear the receptor at their external surfaces. The limiting membrane of the large MVEs is relatively free from receptor. When the large MVEs fuse with the plasma membrane, their contents, the 50-nm bodies, are released into the medium. The 50-nm bodies appear to arise by budding from the limiting membrane of the intracellular vesicles. Removal of surface receptor with pronase does not prevent exocytosis of internalized receptor. It is proposed that the exocytosis of the approximately 50-nm bodies represents the mechanism by which the transferrin receptor is shed during reticulocyte maturation.

Animals↗

In vitro acylation of the transferrin receptor.

In vitro fatty acylation of the transferrin receptor with [3H]tetradecanoate or [3H]tetradecanoyl-CoA has been demonstrated for isolated sheep reticulocyte plasma membranes. Although less than 5% of the receptor was labeled in vitro, the acylated protein could be readily observed after sodium dodecyl sulfate-gel electrophoresis. The acylated transferrin receptor in the reticulocyte membrane was specifically precipitated with a monoclonal antibody and was absent from mature red cell membranes. Incorporation of fatty acid was dependent on ATP, and fatty acid was 5-10 times less effective as an acyl donor than the acyl-CoA derivative, pointing out the strong potential of this reagent for in vitro acylation of membrane proteins. During in vitro maturation of reticulocytes, the receptor is released in vesicles into the incubation medium. Using reticulocytes labeled with [3H]tetradecanoate, it can be shown that the 3H-labeled receptor is transferred from the cells to the vesicles without loss of acyl groups, suggesting that the vesiculation process does not involve deacylation.

Acyl Coenzyme A↗

Degenerated annulus fibrosus of the intervertebral disc contains collagen type II.

Collagen has been isolated and purified from the pronase digest of the outer annulus fibrosus of degenerated and unaffected intervertebral discs obtained at necropsy. The mixture of different collagen types has been purified and fractionated by 2 subsequent DEAE-cellulose chromatographic procedures according to established methods. Collagen chain polymers were then isolated by molecular sieving. Their reduction with 2-mercaptoethanol followed by agarose chromatography resulted in the separation of 3 major peaks corresponding to gamma, beta, and alpha chains. Cyanogen bromide (CNBr) peptides and aminoacid analyses revealed the identity of the isolated alpha-chain as alpha 1(III). Further support for this finding was obtained by immunofluorescence. It is suggested that the presence of collagen type III in the outer annulus fibrosus may be related to intervertebral disc prolapse.

Aged↗

Phosphorylation of the transferrin receptor in isolated sheep reticulocyte plasma membranes.

The transferrin receptor of sheep reticulocyte plasma membranes undergoes phosphorylation with [gamma-32P]ATP in isolated plasma membranes. The phosphorylation is stimulated by Mn2+, Co2+, and Mg2+, but not by Ca2+, Ba2+, Zn2+, Fe2+, or Cu2+. There is no detectable effect of cyclic nucleotides on the phosphorylation of the receptor. Transferrin and a monoclonal antibody against the transferrin receptor have no apparent effect on receptor phosphorylation in intact cells or isolated membranes. Immunoprecipitates of the receptor retain ability to phosphorylate the receptor. The phosphorylation appears to be at a serine residue which turns over with a half time of 20-30 min. ATP appears to be the best, but not the only substrate for receptor phosphorylation.

Amino Acids↗

The fate of the transferrin receptor during maturation of sheep reticulocytes in vitro.

The transferrin receptor of sheep reticulocytes is excised from the cell during in vitro maturation to erythrocytes. The excised receptor may be recovered from the medium by centrifugation at 100 000 X g. Loss of transferrin-binding activity parallels the loss of binding of anti-receptor antibody, as well as RNA content. The released receptor retains the molecular size of the receptor isolated from the plasma membranes (93 000 monomer, 186 000 dimer), has an identical iodotyrosyl peptide map, and is still capable of binding transferrin, as well as an antibody directed against the receptor. The receptor is released in a vesicular form. The major peptides of the vesicles are the receptor and an unidentified peptide of 70 000 whose iodotyrosyl peptide map is distinct from that of the receptor. Although the transferrin receptor has been shown to undergo posttranslational modification (phosphorylation, acylation, and glycosylation) in cultured cells, it has not been established whether any of the transformations are retained in nongrowing cells. In the present communication, it is shown that isolated reticulocyte plasma membranes are capable of receptor phosphorylation, a process previously shown only with intact, cultured cells. The phosphorylating activity is retained in immunoprecipitates of the receptor, but is absent in the vesicles released during maturation. No evidence has been obtained for an effect of either transferrin or an anti-receptor antibody on receptor phosphorylation in intact cells or isolated membranes.

Animals↗

Effect of glycosaminoglycan polysulphate on the metabolism of cartilage ribonucleic acid.

The effect of glycosaminoglycan polysulphate (GAGPS, Arteparon) on the metabolism of cartilage ribonucleic acid (RNA), collagen, and proteoglycan was studied. GAGPS stimulated the synthesis of collagen and the both components of proteoglycan. The effect of GAGPS on RNA was compared with those of chondroitin sulphate and heparin. All three investigated substances enhanced the RNA synthesis, GAGPS, however, had a comparable effect in concentrations ten times smaller.

Animals↗