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Biomedical subjects

W G Thilly

Publications and source records attributed to W G Thilly.

At least 127 records · Page 7Linked to original sources
PubMed1983

High yields from microcarrier cultures by medium perfusion.

A culture perfusion system is described for the growth of anchorage-dependent cells on microcarriers. The critical component of this system is a column separator, which removes medium while allowing the microcarriers to remain in the culture. Amino acids and ammonia were analysed during cell growth of the perfusion culture. None of the amino acids was completely utilized. The accumulation of ammonia to 2.3 mM was observed and may be responsible for, or coincident with, events limiting further cell growth. It is suggested that oxygen deprivation and growth inhibitor accumulation, rather than nutrient depletion, are the major factors in limiting even higher cell yields.

Amino Acids↗
PubMed1983

General approach to the biological analysis of complex mixtures.

The study of potential health effects of combustion effluents involves identifying the substances present and estimating the probable health hazards of each. Unfortunately, this second step cannot be done by using present techniques. Approximations of health hazards by bacterial and human cell assays are being used to set priorities for further biological studies and to suggest needs for modifications of combustion systems. The assumptions underlying this approximation are discussed, and several examples of combustion effluents are reviewed.

Air Pollutants↗
PubMed1982

MDCK microcarrier cultured: seeding density effects and amino acid utilization.

The growth of Madin Darby canine kidney cells on microcarriers was studied at different cell seeding densities. Maximum growth was attained when a cell-to-bead ratio of 7:1 was used. Under these conditions an initial concentration of above 3 X 10(6) cells/ml resulted in a yield of over 2 X 10(6) cell/ml in 2 d. The amino acid utilization of cells from Dulbecco's modified Eagle medium was studied under the above conditions. Eight amino acids (arg, cys, gln, ile, leu, met, ser, and val) showed rapid and near complete depletion from the medium. Five amino acids (his, lys, phe, thr, and tyr) showed limited depletion. Two amino acids (ala and gly) were released into the medium by the cells.

Amino Acids↗
PubMed1982

Selection of mitotic Chinese hamster ovary cells from microcarriers. Cell cycle-dependent induction of mutation by 5-bromo-2'-deoxyuridine and ethyl methanesulfonate.

Large quantities of mitotic cells may be collected by mitotic detachment from a population of Chinese hamster ovary cells growing on positively charged dextran microcarriers in suspension culture. Exponentially growing cells are treated for 2.5 h with colcemid and mitotic cells are detached from the microcarriers by increasing the stirring speed. A yield of 4-6% of the total population is obtained and, of the cells collected, 85-95% are arrested in metaphase. Using this means to synchronize cells we have determined the cell cycle dependence of the toxic and mutagenic effects of 5-bromo-2'-deoxyuridine (BUdR) and ethyl methanesulfonate (EMS). Mutation was measured at two independent loci: resistance to 6-thioguanine and resistance to ouabain. Both mutagens were more toxic during S phase as compared to G1 or G2 or mitosis. BUdR induced significant mutation only during S phase. The maximum induction of 6-thioguanine resistance was observed in cultures treated 10 h after plating of mitotic cells (2 h into S phase), while the maximum induction of ouabain resistance was observed in cultures treated 10-12 h after plating of mitotic cells (2-4 h into S phase). EMS induced significant mutation at all points in the cell cycle. Mutation induction reached a minimum during S phase but the magnitude of difference between any two points in the cell cycle was found to be less than two-fold.

Animals↗
PubMed1982

Mutation assay at the thymidine kinase locus in diploid human lymphoblasts.

A thymidine kinase heterozygote was isolated from a diploid human lymphoblast line which forms colonies with high efficiency in microtiter dishes. We show that this cell line, called TK6, can be mutated from a TK+/- to TK-/- state by diverse mutagens, including ethyl methanesulfonate, butyl methanesulfonate, nitrosomethylurea, UV light, ICR-191, 4-nitroquinoline oxide, fluorodeoxyuridine, benzo[a]pyrene and aflatoxin B1. We report here the experiments required to demonstrate the applicability of this new line in quantitative assays of mutation in human cells. Mitotic recombination between the centromere and the tk locus could not be induced by either dimethylsulfoxide or phorbol-12-myristate-13-acetate.

Cell Line↗
PubMed1982

Mutagenicity of N-nitroso bile acid conjugates in Salmonella typhimurium and diploid human lymphoblasts.

Two N-nitroso bile acid conjugates, N-nitrosotaurocholic acid and N-nitrosoglycocholic acid, were tested for mutagenicity by forward mutation assay in Salmonella typhimurium TM 677 and in diploid human lymphoblasts, line TK6. N-Nitrosoglycocholic acid and N-nitrosotaurocholic acid showed similar concentration-response curves in the bacterial assay with statistically significant mutant fractions observed at about 0.12 mM. Nonnitrosated parent compounds were nonmutagenic. However, in the human cell assay, N-nitrosotaurocholic acid gave statistically significant mutant fractions only at 0.4 mM, but N-nitrosoglycocholic acid was mutagenic at 0.05 microM, some 9000 times more potent. Experiments with quantitative Ames' S. typhimurium reversion assays indicated mutagenesis via base substitution.

Cell Line↗
PubMed1981

Radiosterilization of rat liver microsome containing postmitochondrial supernatant for mutation assays.

Gamma-irradiation was effectively employed to sterilize rat liver postmitochondrial supernatant (PMS), which is required for the metabolic activation of soots and soot-derived polycyclic aromatic hydrocarbons to mutagens. When a known number of Bacillus subtilis spores were added to the PMS and gamma-irradiated at -80 degrees C, a 2-Mrad dose resulted in a 7.5 log kill of the spores. A dose of 3 Mrads was selected as a sufficient effective sterilizing dose and had no significant effect upon the ability of gamma-irradiated PMS to metabolically activate diesel soot and two diesel soot components, benzo(a)pyrene and fluoranthene to mutagens in a Salmonella typhimurium 8-azaguanine resistance forward mutation assay. Three Mrads of gamma-irradiation also had no effect upon the ability of PMS to activate benzo[a]pyrene to a mutagen for the human lymphoblasts. However, gamma-irradiation did reduce the ability of PMS to activate dimethylnitrosamine to a mutagen for S typhimurium.

Animals↗
PubMed1980

Comparison of toxicity and mutagenicity of methylnitrosourea, methylnitronitrosoguanidine and ICR-191 among human lymphoblast lines.

The toxic and mutagenic effects of the alkylating agents methylnitrosourea (MNU) and methylnitronitrosoguanidine (MNNG) and of the frameshift mutagen, ICR-191 were compared among 3 human diploid lymphoblast lines, MIT-2, WI-L2 and GM 130. The MIT-2 and WI-L2 lines were both sensitive to the toxic and mutagenic effects of all 3 agents tested. The WI-L2 line was more sensitive to the toxic effects of MNU and MNNG than the MIT-2 line, while it was somewhat less sensitive to the mutagenic effects of these alkylating agents. The GM 130 line was strikingly resistant to both the toxic and mutagenic effects of the alkylating agents. The order of sensitivity to the toxic effect of ICR-191 was MIT-2 greater than WI-L2 greater than GM 130, while the order of sensitivity to the mutagenic effects of this frameshift mutagen was GM 130 greater than MIT-2 greater than WI-L2. These results point to the importance of accounting possible variations in mutability among individuals when extrapolating from any single mutagenicity assay for human risk assessment.

Aminacrine↗
PubMed1980

Mutagenicity of hydroxamic acids and the probable involvement of carbamoylation.

A series of hydroxamic acids (aceto-, propiono-, benzo-, and p-nitrobenzo-) and seven derivatives of these were examined for biological activity using Salmonella typhimurium. Acylation to yield O-acetyl and O-benzoyl derivatives markedly enhanced toxic properties and was necessary for mutagenic activity for all but p-nitrobenzohydroxamic acid. The dose necessary to produce a minimum significant mutagenic response varied from 21 microM for O-benzoyl benzohydroxamate to 430 microM for O-acetyl acetohydroxamate. These two compounds were also tested with human lymphoblasts to which they were toxic at 100 microM but not mutagenic. O-Acetyl benzohydroxamate, a mutagen, was prepared wih a 14C label in the carbonyl carbon atom of the benzoyl group and was shown to form an adduct in vitro with DNA and polyguanylic acid. The level of binding was 1 mol of 14C per 5 X 10(4) mol of DNA phosphate and 1 mol of 14C per 10(5) mol of polyguanylic acid phosphate.

Cells, Cultured↗
PubMed1979

Human interferon production with diploid fibroblast cells grown on microcarriers.

A variety of diploid human fibroblast lines have been successfully grown to high densities (greater than 10(6) cell/ml) on recently developed microcarriers. Interferon induction using poly I.poly C and a superinduction procedure resulted in yields greater than 10,000 units/ml with one cell line. A direct comparison of microcarrier cultures to roller bottle cultures showed equivalent interferon yields on a per cell basis and some apparent differences relating to optimum inducer concentrations and kinetics of interferon accumulation.

Cell Line↗
PubMed1979

Mutation of human cells by kerosene soot.

The polycyclic aromatic hydrocarbon fraction of a kerosene soot induced forward mutation in human diploid lymphoblasts when coincubated with Sprague-Dawley rat liver postmitochondrial supernatant. Two components of the kerosene soot extract, benzo[a]pyrene (BP) and cyclopenta[cd]pyrene (CP), were also tested. TP was not mutagenic at the concentration found in the soot extract, although it was active at higher concentrations. The amount of CP present could account for approximately 8% of the total mutation observed with the soot. The results were compared to data obtained previously in a similar mutation assay in Salmonella typhimurium. The protocol described permits the facile assay of mutation at the hgprt locus in human lymphoblasts; such mutation is induced by compounds of complex mixtures requiring mixed-function oxygenase activity for metabolism to genetically active derivatives.

Animals↗