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Biomedical subjects

S E Bloom

Publications and source records attributed to S E Bloom.

At least 55 records · Page 3Linked to original sources
PubMed1986

Correlation between induction of xenobiotic metabolism and DNA damage from chemical carcinogens in the chick embryo in vivo.

Chick embryo liver and lung have high levels of basal mixed-function oxidase (MFO) enzyme activities during embryonic development. These activities are differentially inducible by 3,4,3',4'-tetrachlorobiphenyl (TCB) and phenobarbital (PB) in liver but are not inducible in lung. Nucleated chick embryo erythrocytes have no detectable basal or inducible MFO activities. Following induction by TCB or PB, embryos were exposed to the direct-acting carcinogen methyl methane-sulfonate (MMS) or the indirect-acting carcinogens aflatoxin B1 (AFB1) or 7,12-dimethylbenz[a]anthracene (DMBA). DNA damage was measured in liver, lung and blood by the DNA alkaline elution technique. Maximum DNA damage by MMS in liver, lung and blood and by AFB1 and DMBA in liver and lung occurred approximately 3 h after exposure. DNA damage was rapidly repaired in liver and lung, but was not repaired in blood. AFB1 and DMBA caused no detectable DNA damage in blood, even following PB or TCB induction. MMS-induced DNA damage was unaffected by induction in all groups with the exception that PB decreased DNA damage in liver at the low dose of MMS. AFB1- and DMBA-induced DNA damage was increased in liver and decreased in lung following induction by PB and TCB in 14-day embryos. In contrast, AFB1-induced DNA damage was increased by PB and decreased by TCB in 7-day embryonic liver. These results suggest that MFO induction can increase or decrease DNA damage in vivo, but that the direction and magnitude of change in DNA damage can vary depending on the inducer, carcinogen, tissue and the developmental age being investigated.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1985

Embryonic exposure to aflatoxin-B1: mutagenicity and influence on development and immunity.

Chick embryos were mutagenized in ovo in order to study developmentally related alterations in immune functions in survivors of this prenatal toxicant insult. In this experimental system, a single exposure of 6-day chick embryos to 0.1 microgram aflatoxin-B1 (AF-B1) in 10 microliters of acetone was employed, and the control embryos received 10 microliters of solvent alone. This dosage of AF-B1 administered to 6-day embryos was found to increase the incidence of sister chromatid exchanges in blood cells approximately fivefold above the baseline observed in solvent controls. A second sham control, where no solvent was administered, was included in some experiments. The cell cycle times in blood increased slightly during the initial exposure to AF-B1. However, a majority of the AF-B1 and acetone exposed embryos survived and hatched without incident. Losses occurred mainly in the latter part of embryogenesis. After hatching, no significant differences were observed in body weight between different treatment groups up to 26 weeks of age and no change in primary humoral immunity was detected. In contrast, two parameters of cell-mediated immunity, graft vs host (GvH), and cutaneous basophil hypersensitivity (CBH) reactions were both depressed as a result of exposure to AF-B1. The AF-B1 treatment group was significantly reduced in the GvH reaction compared with sham-treated controls. In the CBH assay, AF-B1-exposed chicks showed reduced immunity compared with acetone controls. These results suggest that long-term selective immune depression can occur following embryonic exposure to AF-B1.

Aflatoxin B1↗
PubMed1985

The ribosomal RNA gene cluster in aneuploid chickens: evidence for increased gene dosage and regulation of gene expression.

In the chicken, the nucleolus organizer regions, or sites of the genes encoding 18S, 5.8S, and 28S ribosomal RNA (rRNA), map to one pair of microchromosomes that can be identified by silver nitrate cytochemistry. This nucleolar organizer chromosome also contains the major histocompatibility complex. Chickens aneuploid for this chromosome have been identified and reproduced for over seven generations. Crossing two trisomic parents results in the production of viable disomic, trisomic, and tetrasomic progeny, showing two, three, and four nucleoli and nucleolar organizers per cell, respectively. A molecular analysis of rRNA genes was undertaken to establish the gene copy numbers in the aneuploid genotypes, and to determine if elevated numbers of rRNA genes are stably maintained and inherited over multiple generations. Gene copy numbers were determined using hybridization analysis of erythrocyte DNA obtained from individuals comprising a family which segregated disomic, trisomic, and tetrasomic genotypes. The values obtained were 290, 420, and 570 rDNA repeats per cell for disomic, trisomic, and tetrasomic animals, respectively. These results provide molecular confirmation of the two aneuploid states and show that elevated gene copy numbers have been maintained over multiple generations. Fibroblasts derived from disomic and tetrasomic embryos were found to grow at similar rates in culture, and mature rRNA levels in chicken embryo fibroblasts from disomic, trisomic and tetrasomic embryos were also found to have similar levels of mature rRNA. Therefore, despite the increase in rDNA content, the level of rRNA is regulated to diploid amounts in aneuploid fibroblasts.

Aneuploidy↗
PubMed1984

Evaluation of the genotoxic and embryotoxic potential of chlorpyrifos and its metabolites in vivo and in vitro.

The genotoxicity and embryotoxicity of chlorpyrifos (CPF) and two metabolites were evaluated using the chick embryo, Chinese hamster ovary cells, and by examining blastocysts from superovulated cows crossed to chlorpyrifos-treated bulls. Chlorpyrifos and metabolites were dissolved in acetone and administered to 3-day embryos by the air cell method. The LD50 was 1,500 micrograms/embryo when mortality was checked through and including 17 days of development. The metabolites were more embryotoxic than the parent compound, CPF. Chlorpyrifos and metabolites did not increase the sister chromatid exchange (SCE) frequency above background at any dosage in the 3-day chick embryo assay. Similarly, none of these compounds increased SCE frequencies in three-point dosage tests (1, 10, 100 micrograms/ml) using Chinese hamster ovary cells. Controls in these assays consisted of the solvent carrier acetone (7.0 +/- 2.5 SCE/cell) and 8.6 micrograms/ml methyl methane sulfonate (30.5 +/- 7.4 SCE/cell). Studies of bovine blastocysts obtained from superovulated cows crossed with Dursban 44 treated bulls did not reveal evidence of chromosome aberrations or developmental anomalies associated with pesticide application. However, reproductive performance of breeders may be subnormal as a result of severe poisoning. This underscores the limitations of short-term assays and emphasizes the need to perform thorough toxicological assays of a chemical according to actual usage patterns in the species of concern.

Animals↗
PubMed1984

Correlation between mixed-function oxidase enzyme induction and aflatoxin B1-induced unscheduled DNA synthesis in the chick embryo, in vivo.

The unscheduled DNA synthesis (UDS) technique has been adapted for use in the chick embryo, in vivo, to determine the relationship between induction of the mixed-function oxidase (MFO) enzyme system and genetic damage from an indirect-acting mutagen-carcinogen. Embryos were injected at 6 days of incubation (DI) with either phenobarbital (PB), a specific inducer of P-450-associated enzyme activities, or 3,4,3',4'-tetrachlorobiphenyl (TCB), a specific inducer of P1-450-associated enzyme activities. Aflatoxin B1 (AFB1) was injected 24 hr later (7 DI), followed by a 5-hr continuous 3H-thymidine exposure. The livers were removed, prepared for autoradiography, and hepatocytes were scored for an increase in grains/nucleus, indicative of UDS. Aflatoxin B1 caused a dose-related increase in UDS in all control and induction groups. Phenobarbital-induced embryos had an increased UDS response while TCB-induced embryos had a decreased UDS response, relative to noninduced embryos, for each dosage of AFB1. This suggests that the genotoxicity of an indirect-acting mutagen-carcinogen can be either increased or decreased, in vivo, depending on the inducer used. The chick embryo provides an excellent system for studying the effect of MFO induction on the genotoxicity of promutagen-carcinogens in a developing system.

Aflatoxin B1↗
PubMed1984

Sister chromatid exchange studies in the chick embryo and neonate: actions of mutagens in a developing system.

The embryonic and neonatal periods represent times when the disease process may be initiated as a result of exposure to environmental mutagens and teratogens. We are using the chick embryo/neonate as an experimental system to detect and study the genotoxicity of environmental chemicals in developing tissues and the resultant biological alterations in survivors of perinatal chemical exposure. In vivo bromodeoxyuridine (BrdUrd) labeling of replicating DNA has been employed to measure basal and induced sister chromatid exchanges (SCEs), a candidate cytogenetic endpoint in genetic toxicology testing. Additionally, SCE induction studies with model promutagens have permitted the detection and study of components of the developing mixed-function oxidase (MFO) system of the liver and other organs. The relationship between specific MFO enzyme induction and SCE generation by promutagens has been studied in ovo and using in vitro assays. The in vivo SCE induction potential of 53 compounds, including known mutagens and nonmutagens, was evaluated in the early chick embryo. About 90% of the mutagens induced SCEs; all nonmutagens failed to induce SCE above baseline. Clastogens such as bleomycin did not induce SCE but did cause massive chromosome damage that was easily detected. Gentian violet (GV) is a direct-acting clastogen that did not show any SCE induction. This agrees with the findings from in vitro mutation assays that incorporate rat liver S-9 preparations. Potency for inducing SCE in the chick embryo correlates well with true mutagenic potency, DNA inhibition, and to some extent with carcinogenic activity. Indirect-acting mutagen-carcinogens induced SCEs and also unscheduled DNA synthesis (UDS) in embryonic cells. Biochemical studies revealed that aryl hydrocarbon hydroxylase (AHH) activity develops in the early embryonic liver as it is first formed at 4-5 da of incubation. The level of AHH activity is sufficient to account for the dramatic SCE response. Enhanced SCE induction occurred in older stage embryos, correlating with the increased basal AHH level and enhanced induction capacity of the liver. Modulation of the MFO enzyme system with specific inducers resulted in altered SCE and UDS responses in vivo and in vitro using a chick microsome/chinese hamster ovary (CHO) mammalian cell assay. Viable embryos and neonates have been obtained following exposure to SCE-inducing levels of the mutagen-carcinogen aflatoxin B1 applied at either 6 da or 12 da of development.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

Development of basal and induced aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity in the chicken embryo in ovo.

The development of the hepatic microsomal mixed-function oxidase system was studied to determine the basal level of embryonic enzyme activity and the inducibility of this system throughout growth and differentiation. Chicken embryo livers were assayed for basal and inducible hepatic aryl hydrocarbon hydroxylase (AHHase; designated elsewhere as AHH) activity from the first appearance of the liver as a discrete organ at 5 days of incubation (DI) through day 10 after hatching. In addition, whole-embryo and viscera preparations were assayed at 3 and 4 DI. Basal AHHase activity was equal to or greater than adult levels from 3 DI through hatching in all preparations (approximately 0.3-0.5 nmol/min per mg). A 3-fold increase in basal activity above adult values occurred at hatching. The onset of inducibility in chicken embryo liver between 5 and 6 DI was concomitant with hepatocyte differentiation. A developmental profile of 24-hr 3,4,3', 4'-tetrachlorobiphenyl-induced AHHase activity showed 15- to 30-fold induction over controls from 7 DI through day 10 after hatching, with a maximum of 15 nmol/min per mg at 14 DI and day 1 after hatching, a specific activity greater than 50% greater than maximal induction in the adult. Embryonic AHHase activity was also induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin, 3-methylcholanthrene, beta-naphthoflavone, and sodium phenobarbital. Induction kinetics throughout embryonic development were similar to those reported for the adult chicken and other animals. These findings demonstrate development of a mixed-function oxidase system in very early embryogenesis and then in the liver as it differentiates. Liver AHHase activity is inducible throughout development and perinatally but such activity is under strict developmental regulation. The chicken embryo has adult levels of AHHase activity which would be sufficient to achieve metabolic activation of promutagens/carcinogens before and after hepatocyte differentiation.

Animals↗
PubMed1983

Hematological toxicology following embryonic exposure to aflatoxin-B1.

The influence of embryonic exposure to aflatoxin-B1 (AF-B1) upon the erythroid system of the maturing chicken was examined using a variety of assays. Since the chick embryo is known to possess mixed-function oxidase activity, this animal serves as an excellent model system for studies of human fetal toxicology. AF-B1 (0.1 microgram) was administered to either 6- or 12-day embryos by the air sac method. This level of AF-B1 was highly mutagenic and was found to induce an average of 10.6 sister chromatid exchanges (SCEs) per cell compared with 1.8 SCEs per cell for the acetone control solvent. Despite selection against treated embryos through acute and chronic embryonic toxicity, hatched chicks from AF-B1 treatment groups exhibited erythroid anemia when compared to the acetone controls. Cell count, hematocrit, and hemoglobin concentration were all significantly reduced in the 12-day AF-B1 treatment groups compared with controls. Both sexes were equally affected. While the number of peripheral erythrocytes was reduced following exposure to AF-B1, the differentiation status of erythrocytes was apparently unaltered. Mean cell volume, percentage of circulating reticulocytes, and incidence of an erythroid differentiation marker, chicken fetal antigen, were parameters in which no treatment effects were observed. An apparent maturation effect was noted since adult hematocrits were similar between control and treatment groups. Possible explanations for this age effect are discussed. The ability to detect significant posthatch erythroid toxicity following embryonic exposure to mutagenic levels of AF-B1 suggests the importance of this general approach to perinatal carcinogenic evaluation.

Aflatoxin B1↗
PubMed1982

The longevity of chemically induced sister chromatid exchanges in Chinese hamster ovary cells.

The persistence of the lesions leading to sister chromatid exchanges (SCEs) following acute exposure of Chinese hamster ovary (CHO) cells to direct-acting chemical mutagens was measured. The results revealed that these lesions (and consequently SCEs) are rapidly eliminated from cells. SCE levels fell to near control values by the third or fourth day (six and eight cell cycles, respectively) following exposure of CHO cells to quinacrine mustard (QM) and mitomycin C (MMC). In contrast, cells exposed to methyl methanesulfonate (MMS) showed a small but significant increase in SCE level over control up to and including the fifth day following exposure (approximately ten cell cycles), suggesting that the behavior of these lesions in cells is influenced, at least in part, by the mechanism by which a specific agent interacts with DNA. The possibility that the decline in SCE level was due to the loss of cell populations with high numbers of exchanges was eliminated by the assessment of cloning efficiencies of treated and control cultures.

Animals↗
PubMed1981

Sister-chromatid exchanges: a report of the GENE-TOX program.

The effects of a number of chemicals on sister-chromatid exchange (SCE) frequencies in in vivo and in vitro systems are reviewed. Standardized protocols for future SCE testing in important systems, as well as for evaluation of test results, are presented. Data reported thus far suggest that SCE analysis may prove useful, especially at a secondary level, as a test of mutagenic carcinogens. Strengths and limitations of SCE analysis are summarized as a guide for future evaluation and use of this procedure.

Alkylating Agents↗
PubMed1980

Differential induction of sister chromatid exchanges by indirect-acting mutagen-carcinogens at early and late stages of embryonic development.

To examine the development of drug-metabolizing enzyme systems in the early chick embryo, the procarcinogens, aflatoxin B1 (AF-B1) and 2-acetylamino-fluorene (2-AAF), and the direct-acting carcinogen, ethyl methanesulfonate (EMS) (positive control), were given to embryos; the sister-chromatid-exchange (SCE) technique was used as an indicator of conversion to active mutagenic metabolites. Chick embryos at two stages of incubation (3-day and 6-day) were exposed to the same graded series of dosages of the compounds for a period of 22 hours. All three mutagens increased the frequency of SCE above the control rate of 1,8 SCEs/cell. While a dose-dependent increase in SCE was obtained for both procarcinogens at each age, the mean SCE frequency was significantly higher in the 6-day embryos for each dosage given. In contrast, the direct-acting mutagen, EMS,. gave a reduced level of SCEs at the older age. These results suggest that the ability of early chick embryos to activate promutagens to forms capable of inducing SCE increases as development advances from three to six days of incubation (DI). In the 6-day embryo, the metabolic conversion is enhanced, resulting in a significant increase in the mutagenicity of the test chemicals.

2-Acetylaminofluorene↗
PubMed1980

The effects of short- and long-term exposure of chick embryos to neutral red on the frequency of sister-chromatid exchange.

The chick embryo was used to study the effects of neutral red (NR) on the frequency of sister-chromatid exchange (SCE) in specific tissues exposed to this mutagen for short and long periods as development proceeded. In short-term trials, aqueous NR at doses of 10, 25 and 100 micrograms was injected in 3-day and 6-day embryos. In each case, embryos were also treated with 5-bromodeoxyuridine (BrdU) for a 24-h period (two cell cycles) and harvested at 4 days and 7 days, resp. A long-term exposure (about 8 cell cycles) was achieved by exposing embryos to NR from day 3 to day 7 of incubation. At a NR dose of 25 micrograms, the chronic exposure resulted in a doubling of the rate of SCE (11.4/cell) over that observed in embryos exposed for only 24 h at either days 3-4 (6.0/cell) or days 6-7 (6.0/cell). At 100 micrograms of NR, the same relationship held with SCE rates of 14.2/cell for the chronic exposure versus rates of 8.0/cell (3-4 days) and 6.9/cell (6-7 days). At 10 micrograms of NR, no such accumulation of SCE occurred upon long-term treatment. These results show an enhanced SCE response upon growth of embryonic cells in the presence of NR for several days. This may be the result of the persistence of past lesions with the addition of more lesions upon continued exposure to NR.

Animals↗
PubMed1979

Further study of the genetic toxicity of gentian violet.

The genetic toxicity of gentian violet was studied with the Ames and the Rosenkranz bacterial assays as well as the cytogenetic assays (Chinese hamster ovary cells in vitro in the presence of rat-liver S-9 fractions, the chicken-embryo and mouse-bone-marrow cells in vivo). Gentian violet was found to be toxic but not mutagenic in the Ames assay. However, it was active in the Rosenkranz assay causing reparable DNA damage. The presence of S-9 in the in vitro cytogenetic assay and in the bacterial assays showed that the activity of gentian violet could be reduced or eliminated. In the in vivo assays, gentian violet was not clastogenic and failed to induce sister-chromatid exchanges. However, gentian violet proved to be highly toxic to growing chick embryos at high dosage and depressed mitotic activities in mouse bone marrow after prolonged treatment. Our study suggested that gentian violet can be inactivated by the liver detoxification system. However, it is potentially hazardous to cells that are exposed to the dye directly (e.g. skin epithelium and cell lining of the gastrointestinal tract).

Animals↗
PubMed1977

Distribution of F-bodies, heterochromatin, and nucleolar organizers in the genome of the central mudminnow, Umbra limi.

Due to the very limited amount of knowledge available on the cytochemistry and architecture of fish chromosomes, an extensive banding study was carried out on chromosomes of the central mudminnow, Umbra limi. Through the use of fluorescent staining, C-banding, and silver staining, the chromosomes were characterized, and an idiogram was constructed showing the location of C-band heterochromatin, F-bodies, and the nucleolar-organizer regions. G-banding was attempted but was unsuccessful. Results of fluorescent staining with 33258 Hoechst and quinacrine HC1 raised some interesting questions concerning the cytochemistry of U. limi chromosomes, as well as the specificity of these stains.

Animals↗
PubMed1976

An improved technique for selective silver staining of nucleolar organizer regions in human chromosomes.

A reliable technique for staining human chromosomal nucleolar organizers (NOR's) with silver solutions is described. The NOR's can be selectively stained dark brown by silver solutions leaving the chromosome arms unstained and available for counterstaining with orcein or Giemsa dyes. Unequivocal identification of chromosome pairs bearing NOR's can be achieved using fluorescent banding techniques followed by silver staining. The silver staining procedure for NOR's was simplified and standardized through control of the chemical and physical conditions during silver impregnation and developing.

Chromosomes↗
PubMed1976

Sister chromatid differentiation and exchanges in adult mudminnows (Umbra limi) after in vivo exposure to 5-bromodeoxyuridine.

An in vivo system for the detection of sister chromatid exchange (SCE) in the central mudminnow, Umbra limi, is presented. Sister chromatid differential (SCD) and SCE were demonstrated by fluorescent and Giemsa procedures 5 to 6 days after the fish were injected with 500 mug/g of BrdU. The exchange rate was found to be 2.64 SCEs metaphase in the intestines and 2.42 SCEs/metaphase in the gills. SCE analysis in U. limi should be a useful tool for measuring the mutagenicity of water-borne chemicals.

Animals↗