The effects of experimental administration of low doses of zearalenone on the histology of ovaries in pre-pubertal bitches.

The experiment involved 30 clinically healthy Beagle bitches aged approximately 70 days with an initial body weight of approximately 8 kg. The animals were randomly divided into two experimental groups (EI and EII) and a control group of 10 animals each. Group EI was administered 50 microg of body weight zearalenone/kg per os for 42 days, group EII received 75 microg of body weight zearalenone/kg per os for 42 days, and the control group was administered placeboper os for 42 days. The bitches were ovariectomized at the end of the treatment period for anatomopathological examination. At the same time, peripheral blood samples were collected for endocrinological analyses (17beta-estradiol and progesterone). Administartion of zearalenone particularly higher doses, resulted in the hyperestrogenism degeneration and atrophy of ovarian cells and tissues with accompanying edema and blood extravasation, leading to increased 17beta-estradiol concentrations and an insignificant decrease in progesterone levels.

Polymorphisms of DNA repair genes and risk of squamous cell carcinoma of the head and neck in young adults.

Squamous cell carcinoma of the head and neck (HNSCC) most frequently arise in the epithelial tissues of the upper aerodigestive tract. Patients with HNSCC, aged <45 years are categorized as young adults (YA). They are characterized by more severe form of this disease and often lack of classical, causative risk factors (tobacco smoking, alcohol abusing) in comparison to older (typical) patients (OP). The study purpose was to establish an anticipated protective role of DNA repair genes polymorphisms against cancer-causing agents. It was assumed that the polymorphisms in these genes may have a significant role in the etiology of HNSCC in YA. Studies were carried out on three groups: YA group with HNSCC (n = 90), young healthy group without cancer (YH, n = 160) and OP with HNSCC (n = 205). Three polymorphisms in DNA repair genes were analyzed: XPD ex23: A35931C, XRCC1 ex10: G28152A, and XRCC3 ex7: C18067T. The choice of these genes was connected with their involvement in three different DNA repair pathways. Genotyping was carried out by polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP) technique. Statistical analysis included: calculation of odds ratio (ORs), 95 % confidence intervals (CIs) and p value. There was no significant difference in the distribution of XPD genotypes in YA compared to OP or YH. The XRCC1 AA genotype variant was observed less frequently in HNSCC YA (4.7 %) than in YH and in OP group (17.1 and 10.8 %, respectively). XRCC3 CT genotype variant was observed more frequently in HNSCC YA (61.8 %) than in YH (36.3 %) and this result is statistically significant. This variant was associated with the borderline increased risk of HNSCC development in an early age, however, a similar tendency was not observed in case of double mutated TT variant. The established differences of genotypes distribution do not seem to differentiate substantially YA and OP in head and neck cancer risk.

The effect of low-dose experimental zearalenone intoxication on the immunoexpression of estrogen receptors in the ovaries of pre-pubertal bitches.

Zearalenone is an estrogenic mycotoxin that often contaminates plant material used in the production of feeds for companion animals. Small daily doses of ingested zearalenone--a competitive substrate modulating the activity of enzymes participating in estrogen biosynthesis at the pre-receptor level--can induce subclinical symptoms of hyperestrogenism in bitches. The objective of this study was to determine the effects of low zearalenone doses on the presence of estrogen receptors in the ovaries of pre-pubertal Beagle bitches. The bitches were divided into three groups of 10 animals each: experimental group I--50 microg zearalenone/kg body weight administered once daily per os; experimental group II--75 microg zearalenone/kg body weight administered once daily per os; control group--placebo containing no ZEN administered per os. The animals were ovariorectomized at the end of the experiment, at 112 days of age. Estrogen receptors were detected in ovarian specimens by immunohistochemical methods. The results revealed an absence of estrogen receptors alpha in all groups. In both experimental groups a decrease in the positive response of estrogen receptors beta in specified structures of ovaries was observed. Very low alpha-zearalenol levels probably attested to the slowing down (hypostimulation) of the biotransformation process. Overall, zearalenone intoxication led to hyperestrogenism during a specific developmental stage of pre-pubertal bitches. As regards hormesis, the threshold dose of zearalenone (adaptive capability) was exceeded in the ovaries of experimental group II animals. The results obtained in both experimental groups suggest that long-term exposure to low-dose zearalenone intoxication decreased the degree of estrogen receptors beta staining in particular structures of ovaries in the experimental bitches, which initiated epigenetic modification mechanisms that inhibited ovarian development.

Polymorphisms of the DNA repair genes XRCC1 and ERCC4 are not associated with smoking- and drinking-dependent larynx cancer in a Polish population.

BACKGROUND: Tobacco smoking and alcohol drinking generate oxidative DNA damage and may contribute to larynx carcinogenesis. The X-ray repair cross complementing 1 (XRCC1) and excision repair cross-complementing rodent repair deficiency, complementation group 4 (ERCC4(XPF)) genes are important components of DNA excision repair systems, which repair DNA damage induced by various factors, including tobacco smoking and alcohol. AIM: To investigate the association between the genotypes of the XRCC1-Arg399Gln (rs25487) and ERCC4-Arg415Gln (rs1800067) polymorphisms and smoking- and drinking-related larynx cancer in a Polish population. METHODS: The polymorphisms were determined by PCR-RFLP method in 253 patients with squamous cell carcinoma of the larynx and 253 sex- and age-matched controls. RESULTS: We did not find any association between the investigated polymorphisms and larynx carcinoma, dependent on either smoking or drinking status. No association was found between these polymorphisms and larynx cancer grade, stage or age at diagnosis. CONCLUSIONS: The results indicated that Arg399Gln polymorphism of XRCC1 gene and Arg415Gln polymorphism of ERCC4 gene may not be associated with smoking- and drinking-related larynx cancer in Polish population.

Factors determining sensitivity of prepubertal gilts to hormonal influence of zearalenone.

Among large husbandry animals, swine are the most predisposed to zearalenone (ZEA) intoxication, mainly because cereal is an important component of their diet. Studies performed so far (in vivo, in vitro) suggest that ZEA and its metabolites, which may appear due to ZEA biotransformation (especially alpha-zearalenole; alpha-ZOL), can modify signaling cascades of endogenous sex steroids, through either receptor or non-receptor mechanisms. Of all age groups of swine, immature gilts are particularly predisposed to zearalenone intoxication, as manifested by the occurrence of genital tract tissue dysfunction on exposure to ZEA. The intensity of the adverse effects observed at either systemic or local level in gilts, when compared to sexually mature swine females, suggest that specific age-dependent physiological conditions may exist, which determine the high sensitivity of gilts to exogenous estrogen-like compounds, including ZEA.

The impact of zearalenone on the level of the selected estrogens in blood serum of sexually immature gilts.

The aim of the study was to evaluate the influence of low dose (LOEL - lowest observed effect level) of zearalenone (200 microg/kg b.w.), applied per os for 7 days (short-term intoxication), on sexual behavior, concentration of the examined xenobiotic and its metabolite and selected estrogens in sexually immature gilts: ovariohysterectomised (group D1) and intact (group D2) animals. Clinical signs of oestrus (reddening, oedema and hyperaemia of the vulva and serorhoea from the reproductive tract--lack of standing reflex) were obserwed in group D1 on day 6 and in group D2 on day 4 of the experiment. Laboratory analyses of blood plasma were carried out determine the presence of zearalenone and alpha-zearalenole. They revealed an increase in the level of alpha-zearalenol before the oestrus, decrease in total amount of both examined substances on day when the oestrus appeared and increase in the level of both examined xenobiotics in the post oestrus period together with the higher share of zearalenone. Medium concentrations of estrone and estradiol within the borders of method determination in the majority of periods examined. Higher levels of estrone (32.0 pg/ml) were found on day 4, in the group D2 and estradiol (6.5 pg/ml) on day 6 in the D2 group. The presents study revealed that zearalenone applied per os at LOEL dose causes the incidence of apparent sexual readiness (without standing reflex) in sexually immature gilts with the somatically immature reproductive system.

Preliminary results of monitoring research on zearalenone presence in blood of women with neoplastic lesions in reproductive system.

The aim of the monitoring of zearalenone presence in the blood plasma of women with neoplastic lesions in the reproductive tract was to asses whether the phytosteride is noted in the patients blood and whether the correlation exists between its presence and the incidence of particular neoplasm. The presence of zearalenone or its metabolite--alpha-zearalenole, was noted in 13.51% of the examined women. In 60% of the patients with the confirmed presence of the xenobiotic it was noted in the lowest concentrations. These patients had neoplastic lesions of Carcinoma corpus uteri type.

Zearalenone applied per os provides adverse effects in structure of chosen parts of bitch reproductive system.

Both earlier investigations and complete formulas of pet food for dogs show that professionally prepared pet food is based in 30% on the material of the plant origin. Our own investigations prove that animal feeding stuffs of the plant origin are very often vectors of many undesirable substances (including micotoxins) that are unfavourable for carnivores. Zearalenone is a micotoxin, which frequently occurs among them. It was revealed in pet food in quantities from 5.0-299.5 microg/kg. It is a very dangerous xenobiotic because of its quick and easy absorption in the organism after per os application in monogastric animals. It is also a sterid and despite its plant origin it binds estrogen receptors in the reproductive tract and causes its hypo- and dysfunction. It is clearly visible in young, growing organisms and those during the menopause. In bitches, dysfunctions of the reproduction system, such as: endometritis-pyometra syndrome, prolonged oestrus, ovarian cysts and others are often found. It is supposed that specific for this species hormonal regulation of the reproduction processes based on the long progesterone and prolactin cycles, and on high sensibility to estrogens plays an important role in the etiopathogenesis of these dysfunctions. The application of hormones in bitches in therapeutic and biotechnical (contraception) purposes are the other factors, which should be taken into account in this aspect. There are some suggestions that therapeutic management, unfortunately connected with mistakes in the medical art, is often a cause of pathological condition in a reproductive organ in bitches. However, another reason of these disorders may be that bitches take zearalenone with the commercial pet food. The long-lasting intake of the feed with a low dose of this micotoxin may be the factor, which complicates hormonal regulation of the reproduction processes and is the cause of many disorders. Our own pilotage investigations showed that application of zearalenone at a dose of 200 microg/kg b.w. for 7 days leads to apoptotic-like changes in granule cells in the bitch reproductive system, atresia of follicular cells and oedema and hyperplasia of the oviductal and uterine cells. They could also indicate a follicular phase of the oestrus cycle in bitch. However, it did not take place.

Association of arylamine N-acetyltransferase (NAT1 and NAT2) genotypes with urinary bladder cancer risk.

Arylamines are known bladder carcinogens deriving from tobacco smoke and environmental pollution. Arylamines are metabolised by NAT1 and NAT2 polymorphic enzymes in reactions of carcinogen activation and detoxification. We analysed genetic polymorphisms in both NAT1 and NAT2 genes in 56 bladder cancer patients and 320 healthy patients. Peripheral blood lymphocytes were collected from each subject and genotyped for NAT1 (six alleles) and NAT2 (four alleles) by PCR-RFLP. A weak association between NAT1 and NAT2 genotypes and bladder cancer risk was found when the genotypes were estimated separately (odds ratio OR 1.2, 95%CI 0.7-2.0, and OR 1.3, 95%CI 0.7-1.9, respectively). Almost all NAT1 genotypes possessing at least one "risk" *10 allele were more frequent in the bladder cancer group than in the control group. There was also an increased frequency of "risk" genotypes along with increased cigarette smoking in bladder cancer patients. The coincidence of NAT1-fast/NAT2-slow appears as a potential risk factor for urinary bladder cancer (OR 1.5, 0.8-3.0), as compared with the other genotype combinations.

The protective effect of vitamins C and E against B(a)P-induced genotoxicity in human lymphocytes.

Polycyclic aromatic hydrocarbons (PAH) are a well-characterized group of mutagens and carcinogens. Benzo(a)pyrene [B(a)P], the best known compound in the group, exerts its genotoxic activity following metabolic activation, when it acquires the properties of an electrophilic reagent that is capable of interacting with DNA. Reactive oxygen species (ROS) can remerge during the PAH metabolic activation. Because of their antioxidant activity, vitamins C and E are thought to act as antimutagenic agents. We designed an in vitro protocol to study the potential protective effect of vitamins C and E toward B(a)P-induced DNA damage. In this study, we examined peripheral blood lymphocytes obtained from healthy nonsmoking female volunteers, 22 to 25 years of age. The cells were exposed in vitro to 1 microM B(a)P in the presence of 40 microM or 100 microM of vitamin C or, alternatively, to 30 microM or 100 microM of vitamin E. The B(a)P-induced DNA damage and repair were estimated as the generation and removal of single-strand DNA breaks measured by the alkaline version of the single-cell gel electrophoresis (comet) assay. The protective effect of vitamins C and E was demonstrated when the vitamins were applied simultaneously with or after the B(a)P. The background level of DNA damage in the presence of vitamins C and E was lower than in the system without the vitamins. The experiments were conducted according to various protocol schemes of the vitamin treatment and the results offer additional evidence of the antigenotoxic activity of vitamins C and E. The vitamin activity does not appear to be connected with the steps in metabolic activation or DNA repair. It seems that both vitamins act as competitors of DNA molecule in reaction with the reactive oxygen species.