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Background: There are many studies on the morphology of the liver and its blood vessels in experimental animals, but such studies are lacking in the mole rat (Spalax leucodon). The aim of this paper was a detailed basic study on the topography, morphology, vascular and biliary branching systems of the liver in the mole rat.Materials and Methods: Coloured gelatine and mixture of coloured lead oxide and linseed oil were injection contrast masses used to obtain vascular and biliary branching pattern in the liver. It was revealed that the liver of the mole rat had five lobes (left, quadrate, right medial, right lateral and caudate lobes).Results: The left, undivided lobe was the largest lobe of the liver. The quadrate lobe was divided into two components by a deep notch. The gallbladder, of cylindrical shape, was present and attached to the quadrate lobe. The common bile duct was formed by the union of the left and right hepatic ducts. The pancreatic duct joined the common bile duct before it entered the duodenum. In the present study, only the right medial lobe and quadrate lobe always showed a single lobar artery, portal and hepatic veins. The left lobe showed four lobar arteries, portal and hepatic veins. The caudate lobe with its two processes and the right lateral and medial lobes had different arterial and portal blood supply as well as hepatic and biliary drainage of these lobes. The intrahepatic branches of the proper hepatic artery ran parallel to the branches of the common portal vein in the same lobes of the liver.Conclusions: The results of this study are significant for comparative studies among different species of rodents and other experimental animals. Morphology, vasculature and biliary tract of the liver in the mole rat were similar to that of other experimental animals and identified differences may be related to the adaptation to the mode of life and diet of this rodent. [ABSTRACT FROM AUTHOR]

Diabetes is one of the main health concerns, especially in developed countries. During the last few decades, the percentage of diabetic persons is constantly increasing. Although the genetic factors have a strong influence in the development of diabetes, environmental influence (physical inactivity, inadequate nutrition leading to obesity) also have an important impact. The main objective of this investigation was to evaluate the possible influence of organic sprouted whole wheat grain flourless bread Tonus® (product of Trivit, Becej, Serbia) on DNA damage at various stages of progression through type 2 diabetes mellitus (T2DM). In addition to control (non-diabetic) subjects we analyzed obese, pre-diabetic and diabetic patients, for a total of four experimental groups. All subjects used to eat Mediterranean diet for at least two years before being included in our study. In each of four groups we had five persons practicing the Mediterranean diet but instead of bread they consumed flourless bread Tonus®. The DNA damage was evaluated on peripheral blood mononuclear cells by alkaline single cell gel electrophoresis (Comet) assay at the very beginning (before starting the Tonus® bread diet), and after exactly the three months of consumption of Tonus® bread. Statistical analysis revealed that only in patients with type 2 diabetes, Tonus® bread intake led to decreased DNA damage compared to the level of DNA damage of these patients before they started Tonus® bread diet. We assume that decrease of body weight and hyperinsulinemia caused by Tonus® bread in the diet might be one of the main causes of decreased DNA damage.

Ephedrine, a natural alkaloid from plants of the genus Ephedra, has a chemical structure similar to catecholamines. It is well established that catecholamines (adraneline, noradrenaline and dopamine) cause genotoxic and mutagenic effects. Therefore, the objectives of this investigation were to examine weather ephedrine can exhibit genotoxic effects on isolated human lymphocytes in the Comet assay. Dose-response of human lymphocytes was determined at the concentration range of ephedrine from 0.0005 µM to 500 µM. Three concentrations of ephedrine (1, 50 and 300 µM) which had acceptable cell viability (over 90%) were used for further experiments with inhibitors of DNA reparation (cytosine arabinoside and hydroxyurea). The obtained results showed that ephedrine did not induce DNA damage in isolated human lymphocytes. However, co-treatment of the negative control with DNA repair inhibitors caused a slight but significant increase of DNA damage, due to an endogenous DNA damage. Interestingly, cells treated with ephedrine and DNA repair inhibitors did not express increased DNA damage. On the basis of the obtained results it can be concluded that ephedrine did not exhibit genotoxic effects on isolated human lymphocytes. This result is in accordance with previous investigations showing negative genotoxicological results for ephedrine using bacterial gene mutation test-systems and in vitro cytogenetic analysis. [ABSTRACT FROM AUTHOR]

Lactoferrin (LTF) is a glycoprotein, a member of transferrin gene family which plays an important role in immune mechanisms in the mammary glands of cows. The amount of lactoferrin increases during inflammatory processes and viral infections. The aim of this investigation was to monitor the distribution of lactoferrin gene genotypes and its connection to milk quality and the occurrence of mammary gland diseases in 46 Holstein-Freisian cows of different age (2-7 years) on a farm near Belgrade. DNA was isolated from blood samples, and the polymorphism of lactoferrin gene was deterimined by PCR-RFLP method using the restriction enzyme Eco RI. We found two alelic forms of this gene in cows included in these experiments (A and B) and two genotypes (AA and AB) in a ratio 71.7% to 28.3%. The genotype BB was not found in this sample. In order to determine the degree of differences between genotypes we used discriminant analysis which has shown that there is a statistically significant difference between genotypes AA and AB with respect to productive parameters. When analysed separately, the only parameter which differed significantly (p=0.021) between two genotypes was total milk production. Individuals with observed genotypes are most similar for the amount of milk fat (p=0.271). There is no statistically significant difference in the number of somatic cells in milk samples between the examined genotypes. [ABSTRACT FROM AUTHOR]

Systemic oxidative stress stemming from increased free radical production and reduced antioxidant capacity are common characteristics of obese individuals. Using hydrogen peroxide (H2O2) to induce DNA damage in vitro, in peripheral blood mononuclear cells (PBMCs) from obese subjects and controls, the DNA protective ability of dihidroqercetin (DHQ) and biochaga (B) alone or in combination, were evaluated. The effects of DHQ and B were estimated under two experimental conditions: pre-treatment, where cells were pre-incubated with the substances prior to H2O2 exposure; and post-treatment when cells were first exposed to H2 H2O2, and further treated with the compounds. DNA damage was evaluated using the comet assay. The results of pre- and post-treatment showed a significant decrease in DNA damage produced by H2O2 in the obese group. This decrease was not significant in control group probably due to a small number of subjects in this pilot study. More prominent attenuation was noted in the pre-treatment with DHQ (250 μg/ml). Analysis of antioxidant properties revealed that DHQ's remarkable reducing power, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, and potent∙OH scavenging properties may contribute to strong attenuation of H2O2-induced DNA damage. Also, B showed strong reducing power, DPPH, and ∙OH scavenging ability, while reducing power and DPPH scavenger effects were increased in the presence of DHQ. Conclusively, DHQ and B may reduce H2O2-induced DNA damage in PBMCs from obese subjects when challenged in vitro, and could be valuable tools in future research against oxidative damage-related conditions., (© The Author(s) 2024. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Thymol is a natural essential oil derived from the plant Thymus vulgaris L. It is known to be beneficial for human and animal health and has been used in beekeeping practice against Varroa mite for years. In this study, the genotoxic and antigenotoxic potential of thymol were evaluated on the honey bee ( Apis mellifera L.) continuous cell line AmE-711 for the first time. Using the Comet assay, three increasing concentrations (10, 100, and 1000 µg/mL) of thymol were tested. Negative control (non-treated cells) and positive control (cells treated with 100 µM H 2 O 2 ) were also included. The absence of thymol cytotoxicity was confirmed with the Trypan blue exclusion test. Thymol in the concentration of 10 µg/mL did not increase DNA damage in AmE-711 honey bee cells, while 100 and 1000 µg/mL concentrations showed genotoxic effects. For testing the antigenotoxic effect, all concentrations of thymol were mixed and incubated with H 2 O 2 . The antigenotoxic effect against was absent at all concentrations (10, 100, 1000 μg/mL) tested. Moreover, thymol enhanced the H 2 O 2 -induced DNA migration in the Comet assay. The obtained results indicate genotoxic effects of thymol on cultured honey bee cells suggesting its careful application in beekeeping practice to avoid possible negative effects on honey bees.

Diabetes, a chronic group of medical disorders characterized byhyperglycemia, has become a global pandemic. Some hormones may influence the course and outcome of diabetes, especially if they potentiate the formation of reactive oxygen species (ROS). There is a close relationship between thyroid disorders and diabetes. The main objective of this investigation was to find out whether peripheral blood mononuclear cells (PBMCs) are more prone to DNA damage by triiodothyronine (T 3 ) (0.1, 1 and 10 μM) at various stages of progression through diabetes (obese, prediabetics, and type 2 diabetes mellitus-T2DM persons). In addition, some biochemical parameters of oxidative stress (catalase-CAT, thiobarbituric acid reactive substances-TBARS) and lactate dehydrogenase (LDH) were evaluated. PBMCs from prediabetic and diabetic patients exhibited increased sensitivity for T 3 regarding elevated level of DNA damage, inhibition of catalase, and increase of TBARS and LDH. PBMCs from obese patients reacted in the same manner, except for DNA damage. The results of this study should contribute to a better understanding of the role of thyroid hormones in the progression of T2DM.

The study was designed to evaluate antigenotoxic effect of methanol Teucrium arduini and Teucrium flavum extracts against mitomycin C (MMC)-induced chromosome and DNA damage in vitro . Cytokinesis-block micronucleus (CBMN) and comet assays were used to investigate effect of plant extracts in different concentrations (125, 250, 500 and 1000 µg/mL) on human peripheral blood lymphocytes (PBLs). The obtained results showed that the all tested concentrations of T. arduini and the highest concentration of T. flavum significantly reduced the MMC-induced micronucleus (MN) frequency in comparison to positive control (only MMC). There were significantly negative correlations between the extracts concentrations and MN frequencies (Pearson, r = -0.905, p  = 0.0001 for T. arduini ; r = -0.861, p  = 0.0001 for T. flavum ). The extracts of both plants further lowered the MMC-decreased nuclear division index (NDI) in a dose dependent-manner (Pearson, r = -0.837, p  = 0.001 for T. arduini ; r = -0.598, p  = 0.040 for T. flavum ), but significantly only in the highest concentration (1000 µg/mL). Comet assay showed that extracts reduced MMC-increased genetic damage index (GDI), significantly in the concentrations of 500 and 1000 μg/mL, in comparison with positive control. Based on our results, it can be concluded that methanol T. arduini and T. flavum extracts possess protective proapoptotic and antigenotoxic effect which is indication of their medicinal relevance and use in treatment.

The alkaline comet assay, or single cell gel electrophoresis, is one of the most popular methods for assessing DNA damage in human population. One of the open issues concerning this assay is the identification of those factors that can explain the large inter-individual and inter-laboratory variation. International collaborative initiatives such as the hCOMET project - a COST Action launched in 2016 - represent a valuable tool to meet this challenge. The aims of hCOMET were to establish reference values for the level of DNA damage in humans, to investigate the effect of host factors, lifestyle and exposure to genotoxic agents, and to compare different sources of assay variability. A database of 19,320 subjects was generated, pooling data from 105 studies run by 44 laboratories in 26 countries between 1999 and 2019. A mixed random effect log-linear model, in parallel with a classic meta-analysis, was applied to take into account the extensive heterogeneity of data, due to descriptor, specimen and protocol variability. As a result of this analysis interquartile intervals of DNA strand breaks (which includes alkali-labile sites) were reported for tail intensity, tail length, and tail moment (comet assay descriptors). A small variation by age was reported in some datasets, suggesting higher DNA damage in oldest age-classes, while no effect could be shown for sex or smoking habit, although the lack of data on heavy smokers has still to be considered. Finally, highly significant differences in DNA damage were found for most exposures investigated in specific studies. In conclusion, these data, which confirm that DNA damage measured by the comet assay is an excellent biomarker of exposure in several conditions, may contribute to improving the quality of study design and to the standardization of results of the comet assay in human populations., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

The review summarizes literature data on the DNA-binding, DNA-protecting and DNA-damaging activities of a range of natural human endogenous and exogenous compounds. Small natural organic molecules bind DNA in a site-specific mode, by arranging tight touch with the structure of the major and minor grooves, as well as individual bases in the local duplex DNA. Polyphenols are the best-studied exogenous compounds from this point of view. Many of them demonstrate hormetic effects, producing both beneficial and damaging effects. An attempt to establish the dependence of DNA damage or DNA protection on the concentration of the compound turned out to be successful for some polyphenols, daidzein, genistein and resveratrol, which were DNA protecting in low concentrations and DNA damaging in high concentrations. There was no evident dependence on concentration for quercetin and kaempferol. Probably, the DNA-protecting effect is associated with the affinity to DNA. Caffeine and theophylline are DNA binders; at the same time, they favor DNA repair. Although most alkaloids damage DNA, berberine can protect DNA against damage. Among the endogenous compounds, hormones belonging to the amine class, thyroid and steroid hormones appear to bind DNA and produce some DNA damage. Thus, natural compounds continue to reveal beneficial or adverse effects on genome integrity and provide a promising source of therapeutic activities., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Diabetes represents one of the major health concerns, especially in developed countries. Some hormones such as the stress hormone adrenaline can induce reactive oxygen species (ROS) and may worsen the diabetes. Therefore, the main aim of the investigation was to find out whether peripheral blood mononuclear cells (PBMCs) from normal persons have less DNA damage induced by adrenaline (0.1, 1 and 10 μM) in comparison to PBMCs from obese, prediabetic and diabetic patients. Also, the biochemical parameters of oxidative stress (TBARS, catalase) and lactate dehydrogenase were monitored. It was observed that higher concentrations of adrenaline (1 and 10 μM) induced DNA damage in the obese, prediabetic and diabetic groups. In healthy individuals only the highest concentration of adrenaline caused significant increase in the DNA damage. In summary, total comet score (TCS) comparison has shown significant differences between groups, and DNA damaging effects of adrenaline were most evident in diabetic patients. The results of the biochemical analysis also demonstrate that adrenaline exerts most obvious effects in diabetic individuals which is manifested as significant change of parameters of oxidative stress. In summary, the obtained results demonstrated that diabetics are more sensitive to genotoxic effects of adrenaline and this effect probably resulted from decreased antioxidative defence mechanisms in various stages of progression through diabetes. Therefore, these results could contribute to a better understanding of a role of endocrine factors to damage of cellular biomolecules which could be useful in finding novel therapeutic approaches and lifestyle changes with an aim to lower the possibility of diabetes complications., (Copyright © 2019. Published by Elsevier B.V.)

Harmful effects of elevated levels of catecholamines are mediated by various mechanisms, including gene transcription and formation of oxidation products. The aim of this study was to see whether the molecular mechanisms underlying the damaging action of adrenaline on DNA are mediated by reactive oxygen species (ROS). To do that, we exposed human whole blood cells to 10 μmol L-1adrenaline or 50 μmol L-1H2O2(used as positive control) that were separately pre-treated or post-treated with 500 μmol L-1of quercetin, a scavenger of free radicals. Quercetin significantly reduced DNA damage in both pre- and post-treatment protocols, which suggests that adrenaline mainly acts via the production of ROS. This mechanism is also supported by gradual lowering of adrenaline and H2O2-induced DNA damage 15, 30, 45, and 60 min after treatment. Our results clearly show that DNA repair mechanisms are rather effective against ROS-mediated DNA damage induced by adrenaline.

This study is aimed at analysing biochemical and genetic endpoints of toxic effects after administration of adrenaline. For this purpose, the study was carried out on Wistar rats and three doses of adrenaline were used: 0.75 mg/kg, 1.5 mg/kg, and 3 mg/kg body weight. To achieve these aims, we investigated the effects of adrenaline on catalase (CAT), Cu, Zn-superoxide dismutase (SOD), malondialdehyde (MDA), nitrite (NO 2 -), carbonyl groups (PCC), and nitrotyrosine (3-NT). Total activity of lactate dehydrogenase (LDH), its relative distribution (LDH 1 -LDH 5 ) activity, level of acute phase proteins (APPs), and genotoxic effect were also evaluated. The obtained results revealed that all doses of adrenaline induced a significant rise in CAT activity, MDA level, PCC, NO 2 - , and 3-NT and a significant decrease in SOD activity compared to control. Adrenaline exerted an increase in total activity of LDH, LDH 1 , and LDH 2 isoenzymes. Further study showed that adrenaline significantly decreased serum albumin level and albumin-globulin ratio, while the level of APPs ( α 1 -acid glycoprotein and haptoglobulin) is increased. The micronucleus test revealed a genotoxic effect of adrenaline at higher concentrations (1.5 mg/kg and 3 mg/kg body weight) compared to untreated rats. It can be concluded that adrenaline exerts oxidative and nitrative stress in rats, increased damage to lipids and proteins, and damage of cardiomyocytes and cytogenetic damage. Obtained results may contribute to better understanding of the toxicity of adrenaline with aims to preventing its harmful effects.

The success of antioxidant therapy in hyperthyroidism implies that disease is mediated by oxidative stress, which is known as one of the causing agents of ageing, degenerative diseases, and cancer. The main objective of our study was to determine possible protective effects of methanolic extract of N. rtanjensis in triiodothyronine (T3)-induced DNA breaks of human lymphocytes under in vitro conditions, based upon plant antioxidant capacity related to its phytochemical profile, mainly its polyphenolic complex. The total phenolic and flavonoid content and the antioxidant activity using in vitro 1,1-dyphenyl-2- picrylhydrazyl reagent (DPPH) was determined in methanolic extracts of plant leaves and flowers. The phenolic compound content of 62.73±1.80mg of GaA/g, exhibited solid antioxidant activity (IC 50 = 112.59±0.95μg/ml). The antigenotoxic activity of 0.2, 0.5 and 1.0mg/ml N. rtanjensis methanol extracts mixture with 100µM of T3 was studied in human lymphocytes in vitro using the Comet assay. It is supposed that the antigenotoxicity of N. rtanjensis methanol extracts was caused by high presence of chlorogenic acid, rosmarinic acid and rutin, all known as efficient antioxidant bioactive compounds, which were determined by ultrahigh-pressure liquid chromatograph with MS/MS Mass Spectroscopy (UHPLC/-HESI-MS / MS).

Catechol groups can be involved in redox cycling accompanied by generation of reactive oxygen species (ROS) which may lead to oxidative damage of cellular macromolecules including DNA. The objective of this investigation was to evaluate possible genotoxic effects of a natural catecholamine adrenaline in cultured human lymphocytes using cytogenetic (sister chromatid exchange and micronuclei) and the single cell gel electrophoresis (Comet) assay. In cytogenetic tests, six experimental concentrations of adrenaline were used in a range from 0.01-500 μM. There were no indications of genotoxic effects of adrenaline in sister chromatid exchange and micronucleus tests. However, at four highest concentrations of adrenaline (5 μM, 50 μM, 150 μM and 300 μM) we observed a decreased mitotic index and cell-cycle delay. In addition, in the Comet assay we used adrenaline in a range from 0.0005-500 μM, at two treatment times: 15 min or 60 min. In contrast to cytogenetic analysis, there was a dose-dependent increase of DNA damage detected in the Comet assay. These effects were significantly reduced by concomitant treatment with quercetin or catalase. Therefore, the obtained results indicate that adrenaline may exhibit genotoxic effects in cultured human lymphocytes, most likely due to production of reactive oxygen species., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The thyroid hormones change the rate of basal metabolism, modulating the consumption of oxygen and causing production of reactive oxygen species, which leads to the development of oxidative stress and DNA strand breaks. Olive (Olea europaea L.) leaf contains many potentially bioactive compounds, making it one of the most potent natural antioxidants. The objective of this study was to evaluate the genotoxicity of L-thyroxine and to investigate antioxidative and antigenotoxic potential of the standardized oleuropein-rich dry olive leaf extract (DOLE) against hydrogen peroxide and L-thyroxine-induced DNA damage in human peripheral blood leukocytes by using the comet assay. Various concentrations of the extract were tested with both DNA damage inducers, under two different experimental conditions, pretreatment and posttreatment. Results indicate that L-thyroxine exhibited genotoxic effect and that DOLE displayed protective effect against thyroxine-induced genotoxicity. The number of cells with DNA damage, was significantly reduced, in both pretreated and posttreated samples (P < 0.05). Comparing the beneficial effect of all tested concentrations of DOLE, in both experimental protocols, it appears that extract was more effective in reducing DNA damage in the pretreatment, exhibiting protective role against L-thyroxine effect. This feature of DOLE can be explained by its capacity to act as potent free radical scavenger.

The aim of this investigation was to evaluate the possible protective activity of Agaricus brasiliensis (=A. blazei sensu Murrill) ethanol extract against thymol-induced DNA damage in human lymphocytes. Before we studied the possible interaction of thymol and A. brasiliensis extract, each component was tested in the comet assay. Thymol significantly increased DNA damage in human lymphocytes at higher concentrations (20, 50, 100, 150, and 200 µg/mL), whereas no genotoxic effect of A. brasiliensis ethanol extract was observed. In simultaneous treatment with thymol (200 µg/mL) and A. brasiliensis ethanol extract (50, 100, 150, and 200 µg/mL), the latter failed to reduce a thymol-induced DNA damaging effect regardless of the applied concentrations. To confirm that thymol induces DNA damage via reactive oxygen species, we performed cotreatment with quercetin. Cotreatment with quercetin (100 and 500 µmol/L) significantly reduced DNA damage caused by thymol (200 µg/mL), indicating that thymol exhibits genotoxicity mainly through induction of reactive oxygen species.

Excessive release of stress hormone adrenaline is accompanied by generation of reactive oxygen species which may cause disruption of DNA integrity leading to cancer and age-related disorders. Phenolic-rich plant product dry olive leaf extract (DOLE) is known to modulate effects of various oxidants in human cells. The aim was to evaluate the effect of commercial DOLE against adrenaline induced DNA damage in human leukocytes by using comet assay. Peripheral blood leukocytes from 6 healthy subjects were treated in vitro with three final concentrations of DOLE (0.125, 0.5, and 1mg/mL) for 30 min at 37°C under two different protocols, pretreatment and post-treatment. Protective effect of DOLE was assessed from its ability to attenuate formation of DNA lesions induced by adrenaline. Compared to cells exposed only to adrenaline, DOLE displayed significant reduction (P<0.001) of DNA damage at all three concentrations and under both experimental protocols. Pearson correlation analysis revealed a significant positive association between DOLE concentration and leukocytes DNA damage (P<0.05). Antigenotoxic effect of the extract was more pronounced at smaller concentrations. Post-treatment with 0.125 mg/mL DOLE was the most effective against adrenaline genotoxicity. Results indicate genoprotective and antioxidant properties in dry olive leaf extract, strongly supporting further explorations of its underlying mechanisms of action., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Chromosomal alterations are a feature of both aging and Alzheimer's disease (AD). This study examined if premature centromere division (PCD), a chromosomal instability indicator increased in AD, is correlated with aging or, instead, represents a de novo chromosomal alteration due to accelerating aging in AD. PCD in peripheral blood lymphocytes was determined in sporadic AD patients and gender and age-matched unaffected controls. Metaphase nuclei were analyzed for chromosomes showing PCD, X chromosomes with PCD (PCD,X), and acrocentric chromosomes showing PCD. AD patients, regardless of age, demonstrated increased PCD on any chromosome and PCD on acrocentric chromosomes in both genders, whereas an increase in frequency of PCD,X was expressed only in women. This cytogenetic analysis suggests that PCD is a feature of AD, rather than an epiphenomenon of chronological aging, and may be useful as a physiological biomarker that can be used for disease diagnosis.

Premature centromere division (PCD) represents a loss of control over the sequential separation and segregation of chromosome centromeres. Although first described in aging women, PCD on the X chromosome (PCD,X) is markedly elevated in peripheral blood lymphocytes of individuals suffering from Alzheimer disease (AD). The present study evaluated PCD,X, using a fluorescent in situ hybridization method, in interphase nuclei of frontal cerebral cortex neurons from sporadic AD patients and age-matched controls. The average frequency of PCD,X in AD patients (8.60 +/- 1.20%) was almost three times higher (p < 0.01) than in the control group (2.96 +/- 1.20). However, consistent with previous studies, no mitotic cells were found in neurons in either AD or control brain, suggesting an intrinsic inability of post-mitotic neurons to divide. In view of the fact that it has been well-documented that neurons in AD can re-enter into the cell division cycle, the findings presented here of increased PCD advance the hypothesis that deregulation of the cell cycle may contribute to neuronal degeneration and subsequent cognitive deficits in AD.

Chromosomal involvement is a legitimate, yet not well understood, feature of Alzheimer disease (AD). Firstly, AD affects more women than men. Secondly, the amyloid-β protein precursor genetic mutations, responsible for a cohort of familial AD cases, reside on chromosome 21, the same chromosome responsible for the developmental disorder Down's syndrome. Thirdly, lymphocytes from AD patients display a novel chromosomal phenotype, namely premature centromere separation (PCS). Other documented morphological phenomena associated with AD include the occurrence of micronuclei, aneuploidy, binucleation, telomere instability, and cell cycle re-entry protein expression. Based on these events, here we present a novel hypothesis that the time dimension of cell cycle re-entry in AD is highly regulated by centromere cohesion dynamics. In view of the fact that neurons can re-enter the cell division cycle, our hypothesis predicts that alterations in the signaling pathway leading to premature cell death in neurons is a consequence of altered regulation of the separation of centromeres as a function of time. It is well known that centromeres in the metaphase-anaphase transition separate in a non-random, sequential order. This sequence has been shown to be deregulated in aging cells, various tumors, syndromes of chromosome instability, following certain chemical inductions, as well as in AD. Over time, premature chromosome separation is both a result of, and a driving force behind, further cohesion impairment, activation of cyclin dependent kinases, and mitotic catastrophe, a vicious circle resulting in cellular degeneration and death.

Premature centromere division (PCD) of the chromosome 18 was analyzed by using fluorescent in situ hybridization (FISH) on interphase peripheral blood lymphocytes isolated from six sporadic Alzheimer disease (AD) patients and six healthy elderly controls. Results of FISH analysis revealed that chromosome 18 expressed PCD in 5.18% interphase nuclei of AD patients, and in 2.59% interphase nuclei of age-matched controls (p<0.05). Our study also showed that hypoploidy and hyperploidy frequency for chromosome 18 exhibited a statistically significant increase in the AD group compared to the control one. The increase in spontaneous aneuploidy of chromosome 18 in AD patients which is correlated with PCD shows that deregulation of the time of centromere separation can be considered as a manifestation of chromosome instability leading to aneuploidy.

Thyroid hormones enhance the metabolic rate and the aerobic metabolism favoring oxidative stress, which is accompanied by induction of damage to cellular macromolecules including the DNA. The aim of the present study was to investigate the ability of thyroxine to induce sister chromatid exchange and micronuclei, and to modulate cell-cycle kinetics in cultured human lymphocytes. Eight experimental concentrations of thyroxine were used, ranging from 2 x 10(-9) to 0.5 x 10(-4)M. Treatment with thyroxine increased the frequency of SCE per cell at the higher concentrations (1.5 x 10(-6), 0.5 x 10(-5), 1.5 x 10(-5) and 0.5 x 10(-4)M). On the other hand, there were no significant aneugenic and/or clastogenic effects observed in the cytokinesis-block micronucleus assay. The results show that thyroxine acted as a relatively weak clastogen compared with the positive control N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). In addition to the genotoxic effects, two high concentrations of thyroxine decreased the mitotic index and caused cell-cycle delay. In conclusion, thyroxine exhibited weak clastogenic effects only at high concentrations. Therefore, effects in humans might appear in cases of acute thyroxine overdose.

Insulin is an anabolic hormone that may facilitate development of malignant diseases in various susceptible tissues due to stimulation of mitotic divisions. In this work, an evaluation of mitogenic and genotoxic effects of human recombinant insulin has been performed in cultures of human peripheral blood lymphocytes. Genotoxic effects were studied by the following test systems: (1) in vitro SCE test, and (2) cytokinesis-blocked micronucleus assay. The obtained results indicate that insulin stimulates mitotic division at an optimal concentration of 10(-8) M. On the other hand, insulin has not exhibited genotoxic properties under experimental conditions in this investigation.

The purpose of this study was to determine possible genotoxic and cytotoxic (or mitogenic) effects of high concentrations of oxytocin, active component of Syntocinon in cultures of human peripheral blood lymphocytes. Two test systems were used: (1) analysis of numerical and structural chromosome aberrations, and (2) the in vitro sister chromatid exchange (SCE) test. On the basis of the results obtained it can be concluded that oxytocin does not express any genotoxical properties. Furthermore, the mitotic index did not change significantly.