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F Pacchierotti

Publications and source records attributed to F Pacchierotti.

48 records · Page 3Linked to original sources
PubMed1989

Cytotoxic effects of benzene on mouse germ cells determined by flow cytometry.

Flow cytometric (FCM) DNA content measurements were performed on testicular monocellular suspensions obtained from mice exposed per os to 0, 1, 2, 4, 6, and 7 ml/kg body weight of benzene in order to investigate its cytotoxic action on germ cells. The effects of benzene were measured 7, 14, 21, 28, and 70 d after treatment. Benzene had no effect on testis weight, but FCM analysis showed the relative percentages of some cell subpopulations (tetraploid and haploid cells) to be different from the control pattern, indicating the occurrence of some cytotoxic damage to differentiating spermatogonia. These data demonstrate that spermatogenesis is sensitive to benzene single exposures as evidenced by an altered cell ratio of testicular cell types.

Animals↗
PubMed1989

A cytogenetic approach to evaluate in vivo somatic aneuploidy. Effects of diethylstilboestrol on mouse bone marrow cells.

A cytogenetic approach to estimate in vivo mitotic nondisjunction is proposed, based on chromosomal counting of mouse bone marrow metaphases differentially stained by bromodeoxyuridine (BrdU) incorporation. The method allows the simultaneous assessment of cell cycle delay, aneuploidy and SCE induction. Male mice, implanted with agar-coated BrdU tablets, were injected i.p. with diethylstilboestrol-diphosphate (DES-dp) in the dose range 10-300 mg/kg and killed 18 or 24 h later. To investigate possible sex differences a group of female mice of the same strain and age was injected with 100 mg/kg. As positive controls six males were injected with 1.8 mg/kg of vinblastine (VBL) sulphate. The induction of cell cycle delay was estimated by the relative frequency of first, second and third mitoses after treatment. In spite of a large biological variability, a dose-dependent delay of cellular proliferation kinetics was observed in DES-treated male mice. Treatment with VBL strongly delayed cell cycle progression, according to its antimitotic activity. Hyperploidy was assessed by chromosome counting of second generation metaphases only. After VBL injection, 10.2% of second mitoses were hyperploid, which is a frequency significantly higher than the 0.2% seen in control mice. No significant effect was detected at any DES dose. SCE induction was estimated in the same cells. A significant increase over the control frequency was observed after 200 and 300 mg/kg of DES. By analysis of variance (MANOVA) the dose--effect relationship was fitted by a quadratic model. A sex difference was observed only for spontaneous frequency of SCE with females showing higher levels probably due to their lower weight and relatively higher BrdU in vivo concentration.

Aneuploidy↗
PubMed1988

Nitrilotriacetic acid (NTA) induces aneuploidy in Drosophila and mouse germ-line cells.

The ability of nitrilotriacetic acid (NTA) to induce aneuploidy was studied in the germ line of both Drosophila and the mouse. The Free Inverted X Chromosomes (FIX) genetic system, adopting a brooding scheme, was used to detect induced aneuploidy in Drosophila, and a cytogenetic method based on chromosomal counting in secondary spermatocytes was used in the mouse. In Drosophila a highly significant (P less than 0.001) increase of aneuploidies was produced by NTA (5 x 10(-2) M), which was greater than that produced by colchicine (7.5 x 10(-6) M) and 5-fluorodeoxyuridine (10(-4) M), which were used as positive controls. Brooding effects were observed with NTA, which produced a maximum induction of chromosomal gain in brood I, suggesting a possible stage-specific action during meiosis. The ability of NTA (275 mg/kg body weight) to induce meiotic aneuploidy (hyperhaploidy) also was confirmed in the mouse (P less than 0.001), where all the aneuploidies detected were attributable to treatment of the metaphase I stage.

Acetates↗
PubMed1988

Cytogenetic investigation of chemically-induced aneuploidy in mouse spermatocytes.

This paper discusses a test system in which mouse spermatocytes are analyzed for aneuploidy induction after mice are treated with various agents. Included in this report are methods and procedures of the assay, criteria for determination of aneuploidy induction, considerations for dose-response and stage-specific actions of agents that cause aneuploidy, and finally, advantages and disadvantages of this test system.

Aneuploidy↗
PubMed1988

Meiotic arrest and aneuploidy induced by vinblastine in mouse oocytes.

Young superovulated female mice were injected i.p. with single doses of vinblastine sulfate just before the onset of the first meiotic division. Secondary oocytes, fixed one by one on a slide, were cytogenetically scored. Evidence of the meiotic arresting activity of vinblastine was produced by the observation of increasing frequencies of M1-arrested oocytes and by the presence of undegenerated chromosome sets of first polar bodies. When the first meiotic division could be undertaken chromosome malsegregation occurred with high frequency, both in terms of aneuploidy and polyploidy. M1-blocked and polyploid oocytes have been interpreted as the consequence of irreversible damage to the spindle induced by vinblastine through its binding on tubulin low-affinity sites; this reaction, in fact, causes microtubule crystallization. According to this mechanism, dose-effect relationships of both phenomena show a threshold at 0.45 mg/kg. On the other hand, the incidence of aneuploid oocytes is correlated with meiotic delay, as detected by the delayed degeneration of polar bodies, and increases linearly with dose. Both phenomena are, therefore, stochastic and can be referred to the binding of the chemical on tubulin high-affinity sites, which is known to cause tubulin depolymerization in a colchicine-like way.

Aneuploidy↗
PubMed1987

Meiotic non-disjunction induced by fission neutrons relative to X-rays observed in mouse secondary spermatocytes. II. Dose-effect relationships after treatment of pachytene cells.

(C57B1/Cne X C3H/Cne)F1 male mice were irradiated with single acute doses of 0.4 MeV neutrons (from 0.11 to 0.72 Gy) or 250 kV X-rays (from 0.25 to 3 Gy) and sacrificed 5 days later. Chromosome preparations of secondary spermatocytes, irradiated at the stage of pachytene, were analysed and the incidence of hyper-haploidies and chromosome fragments was recorded. Data on numerical aberrations were fitted by highly significant linear relationships for both types of radiation. A relative biological effectiveness (RBE) value of 5.65 was estimated by the ratio between the slopes of the two regression lines. The same linear fitting was applied to frequencies of cells with fragments, even if in this case other types of functions could not be excluded. An RBE value was estimated in the same way as for numerical aberrations and yielded a comparable figure of 5.23. A significant correlation was also found between the incidence of numerical and structural aberrations, which points to the chromosome itself as the prevalent target for radiation-induced non-disjunction (ND). In addition, the highly significant linearity of the dose-effect relationship observed for the induction of aneuploidies suggests, as the simplest hypothesis, a single-hit mechanism of radiation action, possibly through pre-non-disjunctional damage to the centromeric region, rather than an indirect induction of segregational difficulties after primarily induced chromatid interchanges.

Aneuploidy↗
PubMed1987

Flow cytometric analysis of the effects of 0.4 MeV fission neutrons on mouse spermatogenesis.

(C57Bl/Cne X C3H/Cne)F1 male mice were irradiated with single acute doses of 0.4 MeV neutrons ranging from 0.05 to 2 Gy, and testis cell suspensions were prepared for cytometric analysis of the DNA content 2-70 days after irradiation. Various cell subpopulations could be identified in the control histogram including mature and immature spermatids, diploid spermatogonia and spermatocytes, tetraploid cells and cells in the S-phase. Variations in the relative proportions of different cell types were detected at each dose and time, reflecting lethal damage induced on specific spermatogenetic stages. The reduction of the number of elongated spermatids 28 days after irradiation was shown to be a particularly sensitive parameter for the cytometrical assessment of the radiosensitivity of differentiating gonia. A D0 value of 0.13 Gy was calculated and compared with data obtained after X-irradiation, using the same experimental protocol. In the latter case a biphasic curve was obtained over the dose range from 0.25 to 10 Gy, possibly reflecting the existence of some cell population heterogeneity. RBE values were estimated at different neutron doses relative to the radiosensitive component of the X-ray curve, and ranged from 3.3 to 4, in agreement with data in the literature. Genotoxic effects were monitored 7 days after irradiation by a dose-dependent increase of the coefficient of variation (CV) values of the round spermatid peak, reflecting the induction of numerical and structural chromosome aberrations, and 14 or 21 days after irradiation by the detection of diploid elongated spermatids, probably arising from a radiation-induced complete failure of the first or second meiotic division.

Animals↗
PubMed1984

Nondisjunction induced in mouse spermatogenesis by chloral hydrate, a metabolite of trichloroethylene.

The effects of chloral hydrate (CH), an in vivo metabolite of trichloroethylene, have been evaluated by cytogenetic observations of mouse secondary spermatocytes after ip treatment with 82.7, 165.4, or 413.5 mg/kg bw. Hyper-haploid metaphases have been scored to determine whether previous observations in various nonmammalian organisms about an effect of this drug on the mitotic spindle could be confirmed in mice. At each dose, the frequencies of hyper-haploid cells have been estimated to assess the response of pachytene, preleptotene, premeiotic, and staminal gonial cells. Significant increases above the control value have been observed particularly after treatment of actively dividing gonial cells, confirming the results obtained with the same batch of the drug in a parallel collaborative investigation with Aspergillus nidulans. Thus: a) chloral hydrate has been shown to be effective in inducing nondisjunction in a mammalian system; b) a prevalent action on the mitotic spindle has been confirmed and quantified; and c) the usefulness of parallel investigations with different methods is stressed, particularly to collect information about the mechanisms of induction of nondisjunction events.

Animals↗
PubMed1983

Reciprocal translocations in ageing mice and in mice with long-term low-level 239Pu contamination.

Single intravenous injections of 185 Bq monomeric 239Pu were given to male mice, and the frequency of primary spermatocytes with reciprocal translocations, determined 724 days after treatment, was not significantly different from that of age-matched untreated controls. These old animals showed significantly higher aberration frequencies than young adults. The data therefore show that for low initial activity and very long retention time the possible cytogenetic effects of incorporated nuclide does not change the age-related pattern of increase of spontaneous chromosome aberrations. Considerations of the main variables involved in the induction of cytogenetic effects of incorporated plutonium, based on literature data, indicate that the initial injected activity, the estimated total accumulated average organ dose, and the retention time interact in a complex way; as far as can be seen at present, the effects seem to be dependent mainly on the initial activity at short times after contamination, while the retention time appears to be predominant in the case of long-term observations.

Aging↗
PubMedDate not supplied

Flow cytometry and sizing for routine andrological analysis.

Flow fluorometry and Coulter type sizing analysis of sperm have been applied separately in order to improve human semen analysis. Different methods of sample preparation were evaluated and a protocol involving prestaining pepsin treatment of sperm samples is proposed for fluorometric analysis. The data obtained with fluorometry and sizing analysis result in different kinds of information: Coulter counting allows to automate sperm counting and fluorometry yields more detailed information about normozoospermia and oligozoospermia by determining the proportion of mature spermatozoa and immature germ-cells. These two methods, together with light microscopy, may help to explore the correlation of fertility and pathology of spermatozoa. The aim of these investigations is to yield the preconditions for simultaneous two-parameter analysis of DNA content and cellular size distributions.

DNA↗