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Biomedical subjects

G Quash

Publications and source records attributed to G Quash.

54 records · Page 3Linked to original sources
PubMed1984

In vitro studies on the entry of polyamines into normal red blood cells.

Polyamines are mainly transported in the blood by erythrocytes: Putrescine, spermidine and spermine can be taken up in vitro by red blood cells (RBC); their entry is greater in the presence of serum than in the presence of plasma, and spermine entry is lower than that observed for the two other polyamines. In the presence of serum, the affinity of RBC for spermidine is 30 fold greater than that for putrescine. The majority of RBC polyamines are present in the hemolysate and are not complexed to high molecular weight material. At + 4 degrees C the polyamine uptake is considerably reduced and for putrescine and spermine practically non existent, but it seems that it is internalization rather than binding which constitutes the dependent step. Though intracellular spermidine and spermine levels reflect differences in uptake rather than in outward flux across the cell membrane, the values of putrescine appear to be the resultant of influx and efflux. The presence of specific receptor sites for polyamines visualized by SEM on the surface of RBC using latex-putrescine spheres, confirms the results obtained with labelled polyamines. Therefore, only the understanding of the polyamine repartition inside the blood compartments would permit the clinical use of those molecules as non statistical tumor markers.

Biological Transport↗
PubMed1983

Differences in polyamine availability and insertion into fibronectins released from normal and transformed cells.

1. Fibronectin released from transformed rat kidney cells compared with that released from normal rat kidney cells shows a 50% increase in amino group availability. 2. No such changes were observed in thiol and carboxy group availability or in sialic acid content. 3. The increased amino group availability is not due to a greater polyamine content, which was about 0.04 pmol/mg of protein. 4. Transglutaminase mediated the insertion of spermidine into normal cell fibronectin with linear kinetics. With fibronectin from transformed cells (temperature-sensitive mutant or wild-type), kinetics typical of substrate inhibition were observed. 5. Immunochemical analysis with an anti-polyamine antiserum and an anti-(human fibronectin) antiserum showed that fibronectins from normal and transformed cells react differently. The significance of these results is discussed in the light of changes in the secondary structure between the two fibronectins.

Animals↗
PubMed1982

Diamine oxidase activity in human melanoma cell lines with different tumorigenicity in nude mice.

The activity of diamine oxidase (DO, EC 1.4.3.6.) which converts putrescine into gamma-aminobutyraldehyde in the degradative pathway of polyamine, was studied in 4 human melanoma cell lines, 2 of which produce tumours in greater than 80% of nude mice (M3Dau, M4Beu), whereas the other 2 induce tumours in less than 25% (M1Dor, M2GeB). The activity of DO in these cells varies with the growth rate: 24 h after seeding there is an initial increase in DO activity, followed by a steep decline during exponential growth. At 96 h, when cells reach saturation density, the activity of DO is significantly greater in the highly tumorigenic cell lines than in the poorly tumorigenic cell lines. Kinetic studies show that for the highly tumorigenic lines apparent Km values are 10.6 X 10(-6)M +/- 0.2 (M3Dau) and 14.2 X 10(-6) M +/- 0.6 (M4Beu), whereas for the poorly tumorigenic lines the values are 4.5 X 10(-6) M +/- 0.3. After transplantation into nude mice, the M1Dor cell line, which exhibits a low Km (app.) for DO of which had high Km (app.) value. Km (app.) determination of DO could be an approach for characterizing human melanoma cells differing in their tumorigenic potential in nude mice.

Amine Oxidase (Copper-Containing)↗
PubMed1980

[Immunochemical evidence for the specific binding of putrescine to plasma proteins including fibronectin].

Human plasma contains proteins capable of binding 14C putrescine by the action of Ca++ activated transglutaminase. These proteins have molecular weights from 32 to 220 K and above. One of these (with a molecular weight of 220 K) has been identified as fibronectin by the use of an antifibronectin antiserum. Evidence for a protein with a molecular weight identical to that of fibronectin has been obtained on PAGE analysis of the precipitate formed on incubating human serum with antipolyamine antiserum.

Calcium↗
PubMed1979

Diamine oxidase and polyamine oxidase activities in normal and transformed cells.

1. The activity of diamine oxidase (EC 1.4.3.6) in normal rat kidney cells and in normal rat kidney cells transformed by avian sarcoma virus (B77 strain) growing in tissue culture varies with the stage of growth. There is an initial stimulation of activity by 24h after seeding, followed by a steep decline during exponential growth (48-72h). Enzyme activity decreases even further as the cells reach saturation density (confluence) after 4 days in culture when the activity in normal rat kidney cells is twice as high as that in transformed cells. 2. Differences of about the same order of magnitude are observed between transformed human cells HeLa, HEp2 (a human epithelioid carcinoma) and normal human fibroblasts, in chicken cells between normal myeloblasts and leukaemic myeloblasts, and in rats between biopsy material from normal mammary tissue and 9,10-dimethylbenz[a]anthracene-induced mammary tumours. 3. Polyamine oxidase activity also varies with the growth of transformed rat kidney cells, but shows no significant variation with the growth of normal rat kidney cells between 24 and 96h after seeding. The activity in cells at confluence is from 3- to 5-fold lower in the transformed than in the normal rat kidney cells. 4. A similar 5-10-fold decrease in activity has been found in 9,10-dimethylbenz[a]anthracene-induced mammary tumours in rats and in human oesophageal tumours. 5. Possible reasons for these observations and the contribution of these two enzymes to cellular putrescine concentrations are discussed.

Amine Oxidase (Copper-Containing)↗
PubMed1979

Latex fetuin spheres as probes for influenza virus neuraminidase in productively and abortively infected cells.

Fetuin bound latex spheres do not adhere to the membranes of non-infected cells but adhere to those of cells productively infected by fowl plague virus (FPV Dobson strain). In contrast, asialo fetuin spheres do not attach to the membranes of productively infected cells. Moreover latex fetuin spheres incubated with extracts of productively infected cells and extensively washed are specifically enriched in neuraminidase activity without any trace of haemagglutinin. These observations suggest that viral neuraminidase in the membrane is the site of attachment of the sialic acid moieties of fetuin spheres. These neuraminidase sites are detectable when L cells are productively infected by a mammalian cell adapted mutant of the Dobson strain (FPV-B) but are not detectable on L cells abortively infected by wild type (FPV+). However, even in the abortive system, neuraminidase is synthesised de novo as shown by its labelling with 14C-glucosamine and by its isolation from labelled extracts of infected cells by latex fetuin spheres. These results show that misintegration of viral neuraminidase in the plasma membrane of L cells is a feature of abortive infection of these cells by the Dobson strain of FPV. However the relationship (if any) of this misintegration to abortive infection remains to be established.

Cell Line↗
PubMed1978

The use of a measles latex reagent for the determination of measles antibodies and in a specific test for multiple sclerosis.

Measles virus antigens covalently linked to latex spheres were used for measuring measles-specific antibodies in a direct agglutination test either in microtitre plates or as a rapid slide-agglutination test. The titres were compared to that obtained by conventional assays. The measles-latex spheres were also used as the antigen for a radio-immuno assay. By incorporating a 14C-radioactive marker into the measles-latex spheres their interaction with lymphocytes from multiple sclerosis and control patients was determined. Lymphocytes from multiple sclerosis patients reacted with a higher percentage of beads at low bead/lymphocyte ratios compared with controls, whereas the reverse was found when the ratio of beads was increased.

Antibodies, Viral↗
PubMed1978

The preparation of latex particles with covalently bound polyamines, IgG and measles agglutinins and their use in visual agglutination tests.

Carboxylated latex particles were substituted with side arms terminating in primary amine and hydrazine groups. The particles were coupled to aldehyde groups generated on glycoproteins which were treated with sodium periodate. Particles having the alipathic primary amine putrescine hapten as the sole substituent and particles linked to glycoproteins such as measles agglutinins and IgG were used to detect the presence of the corresponding antibodies or antigens in biological fluids by agglutination tests.

Agglutination Tests↗
PubMed1976

Modification of diamine oxidase activity in vitro by metabolites of asparagine and differences in asparagine decarboxylation in normal and virus-transformed baby hamster kidney cells.

1. The oxidation of putrescine in vitro by pig kidney diamine oxidase (EC 1.4.3.6) was increased in the presence of 2-oxosuccinamic acid and malonamic acid. 2. It was inhibited by 3-aminopropionamide, oxaloacetate and pyruvate. 3. 2-Oxosuccinamate was derived from asparagine in virus-transformed baby hamster kidney (BHK) cells growing in tissue culture. 4. Asparagine was decarboxylated more efficiently by transformed than by normal BHK cells. 5. In BHK cells transformed by polyoma virus (Py BHK), 2-oxosuccinamate is the most likely immediate precursor of the 14CO2 arising from [U-14C]asparagine, and there was some evidence for its formation in an asparagine-dependent clone of BHK cells before and after their transformation by hamster sarcoma virus (respectively Asn- and HSV Asn-). 6. The relationship between 2-oxosuccinamate and pyruvate and the possible roles of these two substances in controlling cellular diamine oxidase activity are discussed.

Alanine↗
PubMedDate not supplied

In vivo antitumor activity of 4-amino 4-methyl 2-pentyne 1-al, an inhibitor of aldehyde dehydrogenase.

4-amino-4-methyl-2-pentyne-1-al (AMPAL), a new irreversible inhibitor of aldehyde dehydrogenase (ALDH) has been assayed for its in vitro and in vivo antitumor activity. In vitro, AMPAL inhibits the proliferation and the ALDH activity of L1210 and RBL5 cell lines. In vivo, AMPAL significantly increases the mean survival time of mice i.p. grafted with leukemia (L1210, P815, MBL2, EL4, RBL5 cell lines) or carcinoma cells (Krebs cell line), without haematopoetic toxicity. No carcinostatic effect was observed against the P388 leukemia and the 3LL Lewis lung carcinoma. A possible relationship between the ALDH isoenzyme activity of the tumor and its sensitivity to AMPAL is discussed in the light of previous reports concerning the role of aldehydes in cell growth control.

Aldehyde Dehydrogenase↗