Chromosome-damaging action of isoniazid and thiacetazone on human lymphocyte cultures in vivo

Summary Antitubercular drugs in general are given in various combinations, one being isoniazid and thiacetazone. In the present study, was evaluated the in vivo chromosome-damaging effects of a combination of these two drugs in 72 h lymphocyte cultures.Chromosome aberrations were significantly increased in the patients treated with INH and thiacetazone as compared with two types of controls: (1) tuberculosis patients before starting the drug treatment and (2) individuals from the general population. The most frequently observed aberrations were chromatid breaks and gaps.It has been shown that individually, isoniazid may not be clastogenic on human chromosomes in therapeutic doses. The effects of thiacetazone on human chromosomes are not known. Consequently, the enhancement in chromosomal aberrations in the drug-exposed patients may be due to a synergistic effect of isoniazid and thiacetazone or to the clastogenic effects of thiacetazone alone.     

Marijuana exposure in vivo and human lymphocyte chromosomes.

Sequential chromosome examinations of peripheral lymphocte cultures were carried out on 21 adult male volunteers who smoked natural blend marijuana cigarettes containing about 1%, 2%, or no delta9-THC. For a limited number of subjects, blood samples from a single venipuncture were cultured independently in two cytogenetic laboratories, and later the slides were exchaged for re-analysis. There were significant differences between laboratories in the absolute break frequencies recorded. These inter-laboratory differences were demonstrated for both techniques of cell culture and metaphase analysis. Neither laboratory found a statistically significant increase in break frequencies asssociated with marijuana smoking. The present study, therefore, failed to detect a measurable effect of marijuana smoking on chromosomal aberrations in subjects experienced in the use of the drug.     

Combined action of gamma irradiation and 5-bromodeoxyuridine on human chromosomes

A combined effect has been studied of 5-bromo-deoxyuridine (BDU) and gamma-radiation on human lymphocytes at the G0 stage. BDU is shown to induce chromatid aberrations irrespective of its radiation. The frequency of chromatid aberrations rises with the BDU concentration. BDU modifies the effect of gamma-radiation on human lymphocytes at the G0 stage, increasing the number of chromosome aberrations. BDU slows down the lymphocyte cell cycle.     

The Clastogenic effect of Pyrimethamine (Daraprim) on human chromosomes in lymphocyte cultures

In this study the clastogenic effect of pyrimethamine (Daraprim), a folic acid antagonist used for the treatment of toxoplasmosis and malaria on human chromosomes, was investigated. Pyrimethamine was added to in vitro lymphocyte cultures at six different concentrations: 0.05 (normal therapeutic dose), 0.1, 0.2, 0.4, 0.8, and 1.6 mg/ml. No proliferation was observed in any of the cultures containing 1.6 mg/ml pyrimethamine. The results of the cytogenetic evaluations show that the frequency of breaks and gaps increase significantly in dose-dependent manner. Thus, pyrimethamine has a clastogenic effect on human chromosomes.Abbreviations PHAM Phytohemagglutinin M - M Metaphase - G Gap - B break - R Rearrangement - NCA Number Chromosome Abnormalities - FA Folic acid - SCE Sister Chromatid Exchange     

Random position of human heterochromatin-bearing chromosomes in first and third mitoses of lymphocyte cultures

Summary The position of chromosomes 1, 9, and 16 in first and third mitoses of lymphocyte cultures was measured in BUdR-labelled air-dried chromosome preparations. No significant deviation from a random distance could be found between the two homologues of each chromosome, either in the first or in the third mitoses after phytohemagglutinin stimulation. Colchicine treatment also had no influence.     

Replication pattern of the X and Y chromosomes in partially synchronized human lymphocyte cultures

The replication pattern of the X and Y chromosomes at the beginning of the synthetic phase was studied in human lymphocyte cultures partially synchronized by the addition of 5-fluoro-2-deoxyuridine (FUdR). The data were evaluated statistically by an analysis of the distribution of silver grain counts over the X and Y chromosomes. —In cells from normal females, one of the X chromosomes began replication later than any other chromosomes of the complement. The short arm of the late replicating X chromosome started replication earlier than the long arm. The telomeric region of the short arm was a preferential site of DNA synthesis at the beginning of replication. —In partially synchronized lymphocyte cultures from a patient with the XXY syndrome, the Y chromosome started replication together with the late replicating X chromosome. The Y chromosome most frequently replicated synchronously with the short arm of the X. The centromeric region of the Y chromosome initiated synthesis before the telomeric region and appeared to replicate synchronously with the telomeric region of the short arm of the X. These findings are discussed with reference to the pairing of the X and Y chromosomes at meiosis.Supported in part by the National Institute of Health Research Grant HD-01979 and National Foundation Birth Defects Research Grant CRCS-40. Dr. Knight was a predoctoral fellow under National Institute of Health Training Program HD-00049-09.     

Action of amsidine on the evolution of lymphocyte cultures and on the structure of human metaphase chromosomes

Chromosomic alterations can be observed after the effect of a antimitotic amsidine on metaphases of cultured human lymphocytes: gaps, breaks, exchange figures, deletions, rings. Moreover we notice, among these cells, particular lines of big sized lymphoplasmocytoid cells, whose rate is abnormally high when compared to reference cultures.     

Effect of isoniazid on structure of human metaphase chromosomes]

Isoniazid gives rise to chromomerization of human bone marrow metaphase chromosomes in vitro or in vivo. The discontinuous appearance of the chromatids seems to be related to their progressive spiralization during the mitotic division. The medicinal substance tested would have the property of revealing the distribution of heterogenous chromatine by showing down its condensation procedure.     

Antigenotoxic effect of lipoic acid against mitomycin-C in human lymphocyte cultures

Antitumor agents are used in therapy against many forms of human cancer. One of these is mitomycin-C (MMC). As with many agents, it can interact with biological molecules and can induce genetic hazards in non-tumor cells. One of the possible approaches to protect DNA from this damage is to supply antioxidants that can remove free radicals produced by antitumor agents. Lipoic acid (LA) is known as one of the most powerful antioxidants. The aim of this study was to investigate antigenotoxic effects of LA against MMC induced chromosomal aberrations (CA), sister chromatid exchanges (SCE) and micronucleus (MN) formation in human lymphocytes. Lymphocytes were treated with 0.2 μg MMC/heparinized mL for 48 h. Three different concentrations (0.5, 1, 2 μg/mL) of LA were used together with MMC in three different applications; 1 h pre-treatment, simultaneous treatment and 1 h post-treatment. A negative, a positive and a solvent control were also included. In all the cultures treated with MMC + LA, the frequency of abnormal cells and CA/cell significantly decreased compared to MMC. Statistically significant reduction was also observed in SCE/cell and MN frequencies in all treatments. These results demonstrated anticlastogenic and antimutagenic effects of LA against MMC induced genotoxicity. LA showed the most efficient effect during 1 h pretreatment. On the other hand, MMC + LA treatments induced significant reduction in mitotic index than that of MMC treatment alone. These results are encouraging that LA can be a possible chemopreventive agent in tumorigenesis in both cancer patients and in health care persons handling anti-cancer drugs.     

Proliferative activity of lymphocyte cultures due to the mutagenic action of thiophosphamide in vivo and in vitro

Thiophosphamide is capable of inhibiting the cell cycle after the treatment of rabbit blood lymphocytes in vivo and in vitro. To assess the proliferative activity of the cells during cultivation, use was made of the index of the mean division number which was experienced by the cells after mutagenic exposure prior to fixation. As the mutagen dose was raised in vivo and in vitro, the mean division number decreased linearly, namely by approximately the same value with an equivalent dose variation. This indicates that thiophosphamide produces the same cytotoxic action both in vivo and in vitro.     

Time-dependent AluI action on human chromosomes

We have analyzed the pattern of AluI digestion over time on human chromosomes in order to monitor the evolution of the in situ enzyme action. Short treatments followed by Giemsa staining produce a G-like banding effect, whereas longer treatments produce a C-like banding pattern. However, when Propidium iodide staining is used, it reveals a uniform bright fluorescence after short AluI digestions and C bands when longer treatments are developed. We propose that C banding is the result of a uniform DNA removal in non centromeric regions taking place after a critical time point, the initial G like banding being produced by changes in the DNA-proteins interactions.     

Effect of elevated temperatures on human lymphocyte chromosomes

The influence of elevated temperatures (38-41 degrees C) on chromosomes of human lymphocytes on different phases of the cell cycle was studied. A high thermosensitivity of chromosomes was demonstrated during (S + G2)-phases of the cell cycle. There was a significant increase in the number of aberrant cells at t greater than 38.5%. The main types of chromosome aberrations were chromatid and chromosome deletions. Cells with 3-5 aberrations and induction of chromosome aberrations due to breaks in the centromere region were noticed at high temperatures (40-41 degrees).     

Expression of fragile sites in human sperm and lymphocyte chromosomes

Summary Sperm and lymphocyte chromosome studies in a normal, fertile male have shown a high degree of coincidence between chromosome lesions and fragile sites in both types of cells. In this donor we also found that some fragile sites expressed in sperm chromosomes coincided with those expressed in lymphocyte chromosomes. These results indicate that the chromosome lesions expressed in sperm do not occur at random and that they are not technical artifacts. The fragility expression in sperm chromosomes could reflect in vivo conditions. The presence in some sperm metaphases of acentric fragments suggests that chromosome fragility can result in the loss of chromosome fragments or give rise to de novo structural rearrangements. However, the incidence of sperm with chromosomal abnormalities observed in this man was within the normal range.