Cytogenetic variability of lymphocytes from phenotypically normal men: influence of age, smoking, season, and sample storage.

A cytogenetic study was conducted on cultured lymphocytes from a group of 60 male volunteers to determine the baseline of chromosomal aberrations in nonchemical workers. Only males were included in the study to avoid any sex effects on the results. Blood samples were collected from each man every 13 w (quarterly) over a period of 12 m. A single batch of culture medium was used for the entire study. The influence of storing the blood samples prior to culture, donor's age, cigarette smoking, and seasonal variation on lymphocyte mitotic index and chromosomal aberration yield was analyzed. A significant decrease in mitotic activity was observed in cultures from samples stored for 3 d at room temperature (22 +/- 1 degree C). Storing of samples at refrigerator temperature (4 +/- 1 degree C) for up to 3 d prior to culture did not affect lymphocyte growth. Although the mitotic index was found to be inversely proportional to the age of the donors, a significant influence of age on total cytogenetic aberrations was not detectable. A group of 15 smokers appeared to have higher number of chromosomal aberrations; however, the difference in mean mitotic activity between lymphocytes of the two groups was not statistically significant. No detectable seasonal influence was found on any chromosomal aberration category except in the number of chromatid gaps. The mitotic indices of the first quarter cultures, on the other hand, showed significant differences from the other three quarters. The chromosomal aberration baseline of the group was not strikingly different from the ones reported by other investigators.(ABSTRACT TRUNCATED AT 250 WORDS)

Supernumerary chromosomal elements in lymphocytes cultured from phenotypically normal human adult males.

In a group of phenotypically normal men there were approximately 0.24% of metaphase lymphocytes with extra chromosomal elements along with the regular 46 chromosomes. They ranged in size from small acrocentric-acentric elements to elements longer than any chromosome arm. These elements have been referred to as supernumerary chromosomal elements. No significant effects due to donor's age, smoking history, season, storage of blood samples prior to culture, or culture medium, were found either in the frequency of supernumerary elements per cell or in the frequency of cells with supernumerary elements. Furthermore, the same subject did not consistently exhibit supernumerary elements. Furthermore, the same subject did not consistently exhibit supernumerary elements when sampled during four successive quarters of the year. Some of these elements in pairs were identified by G-banding technique as translocation chromosomes bearing long arms of chromosome number 2 and presumptive short arms of chromosome 8, acentric long arms of chromosome 4, and iso-acentric chromosomes for the long arms of chromosome 5. Presumably, more than one type of cytogenetic event occurred in their formation. Circumstantial evidence has been presented to show that the means of elimination of these supernumerary elements is a process of chromosomal disintegration.

The incidence of spontaneous cytogenetic aberrations in lymphocytes cultured from normal humans for 48 and 72 h.

Spontaneously occurring cytogenetic aberrations were compared in 48- and 72-h human lymphocyte cultures from a group of 20 normal male volunteers. Mitotic indices for both cultures times were also evaluated. BUdR (5-bromo, 2-deoxyuridine) labelled metaphases from three men were utilized to determine the cell cycle kinetics in the medium used. There was no statistically significant difference between 48- and 72-h cultures among the majority of aberration categories considered. Possibly, the lymphocyte populations at the two different culture times maintained a consistent yield level. Mitotic indices in 72-h cultures were invariably higher than those in 48-h cultures. An overwhelming number of first division metaphases were observed at 48 h of incubation; however, approximately 22% of the lymphocytes did not enter into mitosis until 72 h in culture. A number of difficulties with regard to utilizing BUdR-labelled first division metaphases for cytogenetic analyses are discussed.

Evaluation of culture media for effects on cell cycle kinetics and incidence of chromosomal aberrations in human blood cells.

Peripheral blood samples from 17 apparently healthy male volunteers were set up in duplicate cultures using three commercially available media: Eagle's MEM, RPMI 1640, and TC 199. BUdR (5-bromo,2-deoxyuridine) (10 micrograms/mL) was added to one of the cultures from each person in each medium after 24 h of culture initiation. All cultures were harvested at 72 h of incubation in the presence of colcemid. RPMI 1640 stimulated the highest mitotic activity in both BUdR-treated and untreated cultures. Higher numbers of first division metaphases corresponded with the higher frequency of chromosome-type aberrations in cultures with Eagle's MEM as compared with RPMI 1640 media. On the other hand, higher numbers of chromatid-type aberrations were present in cultures with TC 199 as compared with those with Eagle's MEM. When the chromosome- and chromatid-type aberration data were pooled to score total cytogenetic abnormalities, an influence of the medium was demonstrable. While cultures with Eagle's MEM and TC 199 had the greater number of first division cells, third of subsequent division cells were most prevalent in RPMI 1640 cultures. It is inferred that the length of the cell cycle, the mitotic index, and to some degree the incidence of spontaneous cytogenetic abnormalities are variable attributes of culture media.

A fertility study of male employees engaged in the manufacture of glycerine.

The fertility status of male employees engaged in the production of chlorinated three-carbon compounds at Dow Chemical Company's Texas Division was investigated. Males who had not worked in the production of industrial chemicals for at least five years prior to the study period served as controls. Data were obtained from a reproductive medical history, from hormone determinations, from a physician's examination, and from a semen analysis. Sperm counts and percent normal sperm forms were major study variables. Comparisons were made between the control and the study groups and within the study group only for duration of exposure, strength of exposure and a combination of the two. Specific chemical exposure subgroups within the study group were also compared. Although the participation rate for the exposed group was relatively low (64%), results indicated no detrimental effect on fertility from exposure to chlorinated three-carbon compounds at levels occurring in the specified work environment.

Vinyl chloride cytogenetics.

This report presents cytogenetic findings from a group of 209 workers employed for up to 28 years in the manufacture of vinyl chloride monomer at the Texas Division of Dow Chemical U.S.A. Cytogenetic evaluation results from this group were compared to results found in examination of individuals being considered for employment. Statistical analyses were performed on a group basis for chromatid aberrations, chromosome aberrations and proportion of abnormal cells; no statistical difference of significance was found between the two groups. Comparison of these results with reported studies suggests that the level of cytogenetic aberrations in vinyl chloride workers is probably related to the length and level of exposure, and that risk of adverse genetic effect can be avoided in controlled, minimal-exposure environments.