Micronucleated Cells Research Articles

DNA damage in oral epithelial cells of individuals chronically exposed to indoor radon (222Rn) in a hydrothermal area.

Hydrothermal areas are potentially hazardous to humans as volcanic gases such as radon (222Rn) are continuously released from soil diffuse degassing. Exposure to radon is estimated to be the second leading cause of lung cancer, but little is known about radon health-associated risks in hydrothermal regions. This cross-sectional study was designed to evaluate the DNA damage in the buccal epithelial cells of individuals chronically exposed to indoor radon in a volcanic area (Furnas volcano, Azores, Portugal) with a hydrothermal system. Buccal epithelial cells were collected from 33 individuals inhabiting the hydrothermal area (Ribeira Quente village) and from 49 individuals inhabiting a non-hydrothermal area (Ponta Delgada city). Indoor radon was measured with Ramon 2.2 detectors. Chromosome damage was measured by micronucleus cytome assay, and RAPD-PCR was used as a complementary tool to evaluate DNA damage, using three 10-mer primers (D11, F1 and F12). Indoor radon concentration correlated positively with the frequency of micronucleated cells (r s=0.325, p=0.003). Exposure to radon is a risk factor for the occurrence micronucleated cells in the inhabitants of the hydrothermal area (RR=1.71; 95% CI, 1.2-2.4; p=0.003). One RAPD-PCR primer (F12) produced differences in the banding pattern, a fact that can indicate its potential for detecting radon-induced specific genomic alterations. The observed association between chronic exposure to indoor radon and the occurrence of chromosome damage in human oral epithelial cells evidences the usefulness of biological surveillance to assess mutations involved in pre-carcinogenesis in hydrothermal areas, reinforcing the need for further studies with human populations living in these areas.

230 - Simultaneous Evaluations of GSH Levels, SOD Activity and the Genotoxic Damage in Mice Treated with Chromium(VI) and Catechins of Green Tea [(+)-Catechin and (-)-Epigallocatechin-3-Gallate

The compounds of Cr(VI) generates reactive oxygen species (ROS) during its reduction to Cr(III) leading to DNA damage. The green tea polyphenols can protect against the genotoxic damage induced by Cr(VI) by reducing frequencies of micronucleus MN[1]. In this study, the effect of catechins of green tea [(+)-catechin and epigallocatechin-3-gallate (EGCG)] on glutathione (GSH) levels and superoxidodismutasa (SOD) activity was evaluated simultaneously with the analysis of MN and apoptosis. Groups of five Hsd-ICR mice were divided and treated as follows: i) only vehicle; ii) (+)-catechin (20 and 40 mg/kg) or EGCG (10 mg/kg) by gavage; iii) CrO3 (20 mg/kg) intraperitoneally; and iv) (+)-catechin or EGCG in addition to CrO3. Evaluations were performed at 0, 24, 48 and 72 h after treatments. No significant changes in MN frequencies were observed when the catechins of green tea were administrated alone. However, the treatment with CrO3 showed an increase in MN, apoptosis and GSH levels, but the activity of SOD decreased. The treatment with (+)-catechin prior to administration of CrO3 showed no effects on MN and apoptosis. However, treatment with EGCG possibly presents both effects; prooxidant and antioxidant because the MN was increased at 24 h, but it was reduced at 48 h when EGCG was administrated prior to CrO3. Additionally, GSH levels decreased and SOD activity is recovered. The increase of apoptotic cells suggests that this may play a role in the elimination of micronucleated cells. Financial support was obtained from DGAPA-UNAM PAPIIT IN219216; IN217712; PAEP-2016; CONACYT 703847/584686. Reference [1] Maria del Carmen Garcia-Rodriguez, et al. 2013. Oxid Med Cell Longev, vol. 2013, Article ID 486419, 9 pag. doi:10.1155/2013/486419.

Potentially harmful elements in house dust from Estarreja, Portugal: characterization and genotoxicity of the bioaccessible fraction.

Due to their behavioral characteristics, young children are vulnerable to the ingestion of indoor dust, often contaminated with chemicals that are potentially harmful. Exposure to potentially harmful elements (PHEs) is currently exacerbated by their widespread use in several industrial, agricultural, domestic and technological applications. PHEs cause adverse health effects on immune and nervous systems and can lead to cancer development via genotoxic mechanisms. The present study is an integrated approach that aims at assessing the genotoxicity of bioaccessible PHEs following ingestion of contaminated house dust. A multidisciplinary methodology associating chemical characterization of five house dust samples, extraction of the bioaccessible PHEs in gastric extracts by the unified BARGE method, determination of the bioaccessible fraction and in vitro genotoxicity of gastric extracts in adenocarcinoma gastric human (AGS) cells was developed. The five gastric extracts induced dose-dependent genotoxicity in AGS cells. Copper (bioaccessible concentration up to 111mg/kg) was probably the prevalent PHE inducing primary DNA damage (up to 5.1-fold increase in tail DNA at 0.53g/l of gastric extract). Lead (bioaccessible concentration up to 245mg/kg) was the most prevalent PHE inducing chromosome-damaging effects (r=0.55; p<0.001 for micronucleated cells induction). The association of principal component analysis and Spearman's correlations was decisive to understand the chromosome-damaging properties of the bioaccessible PHEs in AGS cells. This methodology could be used on a larger-scale study to provide useful information for science-based decision-making in regulatory policies, and a better estimation of human exposure and associated health risks.

Effect of CYP3A4 genetic polymorphisms on the genotoxicity of 4,4′-methylene-bis(2-chloroaniline)-exposed workers

ObjectivesWe investigated the relationship between 4,4’-methylene-bis(2-chloroaniline) (MBOCA) exposure and micronucleus (MN) frequency, and how this association was affected by genetic polymorphism of the cytochrome P450 enzyme (CYP3A4).MethodsWe divided the study...

Genotoxicity, mutagenicity and cytotoxicity of carotenoids extracted from ionic liquid in multiples organs of Wistar rats

The ionic liquid or melted salt 1-Butyl-3-methylimidazolium is an alternative process to extract natural pigments, such as carotenoids. Lycopene represents 80–90% of total of carotenoids presents in tomatoes and it has been widely studied due its potent antioxidant action. The aim of this study was to evaluate genotoxicity, mutagenicity and cytotoxicity of carotenoids extracted from ionic liquid using experimental model in vivo. For this purpose, a total of 20 male Wistar rats were distributed into four groups (n=5), as follows: control group; received a corresponding amount of corn oil for 7days by intragastric gavage (i.g.), ionic liquid group, received 10mgkg−1 body weight for 7days by gavage; 10mg carotenoids group, received 10mgkg−1 bw dissolved in corn oil for 7days by gavage and 500mg carotenoids group, received 500mgkg−1 bw dissolved in corn oil for 7days by gavage. Rat liver treated with ionic liquid exhibited moderate histopathological changes randomly distributed in the parenchyma, such as cytoplasmic eosinophilia, apoptotic bodies, inflammatory infiltrate and focal necrosis. DNA damage was found in peripheral blood and liver cells of rats treated with ionic liquid or carotenoids at 500mg. An increase of micronucleated cells and 8-OhDG immunopositive cells were also detected in rats treated with carotenoids at 500mg. In summary, our results demonstrate that recommended dose for human daily intake of carotenoids extracted by ionic liquid did not induce genotoxicity, mutagenicity and cytotoxicity in multiple organs of rats.

Cyto- and genotoxic potential of water samples from polluted areas in Kosovo.

Reports on the state of the environment in Kosovo have emphasized that river and ground water quality is affected by pollution from untreated urban water as well as the waste water from the industry. One of the main contributors to this pollution is located in Obiliq (coal power plants). Prishtina-the capital city of Kosovo-is heavily influenced too. Furthermore, the pollutants combined together with those from heavy traffic are dissolved in Prishtina runoff water, which is discharged into the creek entering the river Sitnica together with urban waste water. The available data show the complex pollution with excessive quantities of nitrites, suspended materials, organic compounds, detergents, heavy metals, polychlorinated biphenyls, etc. In this study, the cytotoxic and genotoxic potential of water samples taken at these sites was tested in primary rat hepatocytes. The results obtained indicate that water samples collected in Prishtina and Obiliq had a significant cytotoxic potential in primary rat hepatocyte cultures even when diluted to 1%. The increased cytotoxicity, however, was not accompanied by an increased genotoxicity as measured by the percentage of micronucleated cells. Further investigations addressing the chemical composition of the samples and the identification of the toxicants responsible for the cytotoxic effects found will be carried out in a next step.

Evaluation of the Protective Effect of Brassica oleracea (L. var. acephala) in Rats with Surgically-Induced Gastroesophageal Reflux Disease

Background: Gastroesophageal reflux disease (GERD) is a common disorder that may result in esophageal cancer. Although proton pump inhibitors are the standard treatment for this illness, Brassica oleracea may provide new therapeutic possibilities. Objectives: We aimed to evaluate and compare the effects of the B. oleracea extract and the proton pump inhibitor omeprazole on esophageal complications arising from a surgically-induced GERD model in rats. In addition, we investigated possible associations between the frequency of DNA damage and esophageal histological alterations, as well as the genotoxic/anti-genotoxic and cytotoxic/anti-cytotoxic effects of B. oleracea and omeprazole. Methods: Rats with and without GERD were equally divided into groups to receive one of two the treatments, B. oleracea extract (500 mg/kg bw) or omeprazole (30 mg/kg bw), daily over the course of four weeks. A group of non-treated rats received water in the same circumstances. Micronucleus assay was used to assess DNA damage in blood and bone marrow cells. Results: Rats with GERD developed esophagitis and esophageal squamous cell carcinoma. B. oleracea and omeprazole-treated GERD rats presented significantly decreased inflammation in relation to non-treated GERD rats (P < 0.05). However, in rats without GERD, omeprazole significantly increased the frequencies of micronuclei and micronucleated cells as compared to the corresponding cell counts in non-treated rats (P = 0.04). Conclusions: B. oleracea demonstrated similar anti-inflammatory properties to omeprazole in rats with GERD. However, omeprazole also demonstrated genotoxic and cytotoxic effects in rats without GERD.

“Brazilian ginseng” (Pfaffia glomerata Spreng. Pedersen, Amaranthaceae) methanolic extract: cytogenotoxicity in animal and plant assays

Pfaffia glomerata (Spreng) Pedersen is a medicinal plant largely used as an adaptogenic herb. The species is naturally found in Africa and Americas and is the subject of pharmaceutical and commercial interest, mainly due to the accumulation of β-ecdysone in its roots. Brazil is considered the greatest supplier of P. glomerata in the world. Due to the morphological similarity of its roots to those of Panax ginseng (Korean ginseng), the species became known as the Brazilian ginseng. In this study, we performed the cytogenotoxic evaluation of P. glomerata root extract in rodent bone marrow micronucleus and Allium cepa assays. Taking into consideration the therapeutic dosage of P. glomerata, three methanolic extract dilutions were used in the Micronucleus assay. Each animal was treated with two consecutive doses with a 24-h interval. To assess the cytogenotoxicity of the extract, 2000 polychromatic erythrocytes per animal were analyzed. For the Allium cepa assay, onion seeds were exposed during 24h to six methanolic extract dilutions. Cell cycle and chromosomal alterations were quantified. The results of the Micronucleus assay showed genotoxic effects for all treatments investigated, considering the increase in the number of micronucleated cells. In the A. cepa assay the methanolic extract induced alterations in the cell cycle. Aneugenic and clastogenic effects were observed. The results indicate cytogenotoxic activity of P. glomerata extract in animal and plant cells. Thus, the ingestion of P. glomerata for medicinal purposes should be done carefully. The results also lead to the possibility of investigating the potential of the species in the research of prospective compounds for cancer treatment and for isolation of biologically active substances.

Multi-level biological responses in Ucides cordatus (Linnaeus, 1763) (Brachyura, Ucididae) as indicators of conservation status in mangrove areas from the western atlantic

There is a global lack of knowledge on tropical ecotoxicology, particularly in terms of mangrove areas. These areas often serve as nurseries or homes for several animal species, including Ucides cordatus (the uçá crab). This species is widely distributed, is part of the diet of human coastal communities, and is considered to be a sentinel species due to its sensitivity to toxic xenobiotics in natural environments. Sublethal damages to benthic populations reveal pre-pathological conditions, but discussions of the implications are scarce in the literature. In Brazil, the state of São Paulo offers an interesting scenario for ecotoxicology and population studies: it is easy to distinguish between mangroves that are well preserved and those which are significantly impacted by human activity. The objectives of this study were to provide the normal baseline values for the frequency of Micronucleated cells (MN‰) and for neutral red retention time (NRRT) in U. cordatus at pristine locations, as well to indicate the conservation status of different mangrove areas using a multi-level biological response approach in which these biomarkers and population indicators (condition factor and crab density) are applied in relation to environmental quality indicators (determined via information in the literature and solid waste volume). A mangrove area with no effects of impact (areas of reference or pristine areas) presented a mean value of MN‰<3 and NRRT>120min, values which were assumed as baseline values representing genetic and physiological normality. A significant correlation was found between NRRT and MN, with both showing similar and effective results for distinguishing between different mangrove areas according to conservation status. Furthermore, crab density was lower in more impacted mangrove areas, a finding which also reflects the effects of sublethal damage; this finding was not determined by condition factor measurements. Multi-level biological responses were able to reflect the conservation status of the mangrove areas studied using information on guideline values of MN‰, NRRT, and density of the uçá crab in order to categorize three levels of human impacts in mangrove areas: PNI (probable null impact); PLI (probable low impact); and PHI (probable high impact). Results confirm the success of U. cordatus species’ multi-level biological responses in diagnosing threats to mangrove areas. Therefore, this species represents an effective tool in studies on mangrove conservation statuses in the Western Atlantic.

Long-term genotoxic effects of immunosuppressive drugs on lymphocytes of kidney transplant recipients

Immunosuppressive therapy can prevent rejection after organ transplantation. However, increased cancer risk is a serious complication among patients undergoing such therapy. We have evaluated whether prolonged use of immunosuppressive drugs is genotoxic. DNA instability was assessed, using the comet and micronucleus assays, in blood lymphocytes of 76 kidney transplant patients. DNA damage detected by the comet assay increased with time after transplantation. The estimated glomerular filtration rate of the patients did not influence the incidence of DNA damage. No association between micronucleated mononucleated cells and time elapsed after transplantation was observed. Our results suggest that prolonged use of immunosuppressive drugs in kidney transplant patients can induce genetic instability.

Vitamin D insufficient levels during pregnancy and micronuclei frequency in peripheral blood T lymphocytes mothers and newborns (Rhea cohort, Crete)

Vitamin D deficiency is common among pregnant women and may be associated with several adverse health outcomes including cancer. Micronuclei frequency is a biomarker of early genetic effects and has been used to examine the association between genotoxic exposures and cancer. We examined maternal vitamin D levels during pregnancy in associations with micronuclei frequency in maternal blood and in cord blood. 173 mothers and 171 newborns born between 2007 and 2008 in Heraklion (Crete, Greece) were included in the study. Between 14th and 18th weeks of gestation we collected information on maternal diet using food frequency questionnaires (FFQs). We measured maternal serum concentrations of 25-hydroxyvitamin D [25(OH)D] between the first and second trimester of pregnancy. We estimated dietary vitamin D intake using information from FFQ. After delivery we collected cord blood and maternal peripheral blood. We used the cytokinesis-block micronucleus (CBMN) assay to assess the frequencies of micronucleated cells in binucleated T lymphocytes (MNBN). Maternal insufficient serum levels of 25(OH)D (<50nmol/L) during pregnancy were associated with increased MNBN frequency in cord blood [IRR=1.32 (95%CI: 1.00, 1.72)]. This increase was higher for newborns with birth weight above the third quartile [≥3.500kg; IRR=2.21 (1.26, 3.89)]. Similarly, low levels of dietary vitamin D were associated with increased MNBN frequency in cord blood [middle tertile IRR=1.08 (0.78, 1.47), lower tertile IRR=1.51 (1.06, 2.14)]. Insufficient levels of vitamin D were not associated with MNBN in mothers. Our results suggest that vitamin D deficiency during pregnancy increases genotoxic risks in newborns. The prevalence of vitamin D deficiency globally is high and it is important to further investigate whether vitamin D supplementation or similar interventions during pregnancy could prevent DNA damage at early stages of life.

Cytotoxic, mutagenicity, and genotoxicity effects of guanylhydrazone derivatives

Several studies have reported that guanylhydrazones display a variety of desirable biological properties, such as antihypertensive, antibacterial, and antimalarial behaviour. They furthermore promote anti-pneumocystosis and anti-trypanosomiasis, exhibit antitumor activity, and show significant cytotoxicity against cancer cell lines. In this work, we have evaluated the cytotoxicity, mutagenicity, and genotoxicity of two guanylhydrazones derivatives, (E)-2-[(2,3-dimethoxyphenyl) methylene] hydrazine carboxymidamide hydrochloride (2,3-DMeB) and (E)-2-[(3,4-dimethoxyphenyl) methylene] hydrazine carboxymidamide hydrochloride (3,4-DMeB), in different biological models. Both 2,3-DMeB and 3,4-DMeB induce weak cytotoxic and mutagenic effects in bacteria and yeast. The genotoxicity of these compounds was determined in a fibroblast cell line (V79) using alkaline comet assay, as well as a modified comet assay with bacterial enzymes formamidopyrimidine DNA-glycosylase (FPG) and endonuclease III (EndoIII). Both guanylhydrazone derivatives induced DNA damage. Treatment of V79 cells with EndoIII and FPG proteins demonstrated a significant effect of 2,3-DMeB and 3,4-DMeB with respect to oxidized bases. In addition, the derivatives induced a significant increase in the frequency of micronucleated cells at high doses. The antifungal and anti-trypanosomal properties of these guanylhydrazone derivatives were also evaluated, and the obtained results suggest that 2,3-DMeB is more effective than 3,4-DMeB. The biological activity of 2,3-DMeB and 3,4-DMeB may thus be related, at least in part, to their oxidative potential, as well as to their ability to interact with DNA. Considering the previously reported in vitro antitumor activity of guanylhydrazone derivatives in combination with the lack of acute toxicity and the fact that DNA damage is only observed at high doses should render both compounds good candidates for in vivo studies on antitumor activity.

Pure anatase and rutile + anatase nanoparticles differently affect wheat seedlings

TiO2-nanoparticles (TiO2-NPs) are increasingly released to the environment. The present work investigates the cytotoxicity, genotoxicity and uptake of TiO2-NPs in Triticum aestivum. Wheat seeds were exposed to 5–150 mg L−1 of anatase (ana) or rutile + anatase (rut + ana) TiO2-NPs for 5 d. After exposure, germination and growth rates were determined. Cytotoxic effects were evaluated by changes in the cell cycle dynamics and in the membrane integrity. Genotoxicity was assessed by ploidy mutations and DNA-damage, and by mitotic abnormalities. NP uptake was analyzed by Energy Dispersive X-ray Spectroscopy (EDS). Ana-TiO2 revealed higher toxicity regarding the rate of germination, but no negative effects were detected concerning growth. Although roots and shoots showed no EDS-detectable levels of Ti, despite cyto- and genotoxicity was observed in ana and rut + ana-NPs exposed roots. Cell cycle profile was formulation dependent with rut + ana presenting a higher capability to induce a cell cycle arrest at G0/G1. Both formulations induced genotoxic effects by increasing micronucleated cells: for rut + ana a dose-dependent response is evident and seems to be more genotoxic than ana at lower concentrations. Rut + ana also increased membrane permeability. The observed higher cytotoxicity of rut + ana may be explained by the higher photoactivity of this mixture. Overall, these data indicate that during germination, TiO2-NPs induce severe cyto/genotoxic effects, which are dependent on the TiO2-NP formulation.

DNA damage induced by occupational and environmental exposure to miscellaneous chemicals

Epidemiological studies for hazardous situations resulting from the risk of environmental and/or occupational exposure to miscellaneous chemicals present several difficulties. Biomonitoring of human populations can provide an early detection system for the initiation of cell dysregulation in the development of cancer, which would help develop an efficient prevention program. Recently, the cytokinesis-block micronucleus (CBMN) assay in lymphocyte cells has become an important tool for assessing DNA damage in exposed populations. This is the method of choice for population-based studies of occupational and/or environmental exposure to different agents. In this review, human populations exposed to coal, dyes, paints, organic solvents in a complex mixture, and others miscellaneous chemicals were analyzed. Data from 28 studies was evaluated in relation to the effect of complex mixture exposition on micronucleus (MN) frequency. Other biomarkers and the background factors were evaluated as well, such as gender, age, or smoking habit. Most of these studies (75%) showed a significant increase of micronucleated cells to exposed groups in relation to the control groups, besides chromosomal aberrations (CA), sister chromatid exchanging (SCE) and comet cells (comet assay). The studies from this review about miscellaneous chemicals exposures using CBMN assay have indicated some time and dose-dependent effects. Overall, the findings suggest that the responses resulting from exposure to complex mixtures are varied and complicated. However, they are also an important mechanism of DNA damage concerning disruption of metal ion homeostasis that may lead to oxidative stress, a state where increased formation of reactive oxygen species (ROS) overwhelms body antioxidant protection and subsequently could induce cancer.

Acute crack cocaine exposure induces genetic damage in multiple organs of rats.

Crack cocaine is a very toxic product derived from cocaine. The aim of this study was to evaluate genetic damage in multiple organs of rats following acute exposure to crack cocaine. A total of 20 Wistar rats were distributed into four groups (n = 5), as follows: 0, 4.5, 9, and 18mg/kg body weight (b.w.) of crack cocaine administered by intraperitonealroute (i.p.). All animals were killed 24h after intraperitoneal (i.p.) injection. The results showed that crack cocaine increased the number of micronucleated cells in bone marrow cells exposed to 18mg/kg crack cocaine (p < 0.05). Peripheral blood and liver cells presented genetic damage as depicted by single cell gel (comet) assay at 9 and 18mg/kg doses (p < 0.05). Immunohistochemistry data revealed significant increase in 8-hydroxy-20-deoxyguanosine (8-OHdG) immunoexpression in hepatocytes of animals exposed to crack cocaine at 9 and 18mg/kg (p < 0.05) when compared with negative controls. Taken together, our results demonstrate that crack cocaine is able to induce genomic damage in multiple organs of Wistar rats.

The Influence of C-Ions and X-rays on Human Umbilical Vein Endothelial Cells.

Damage to the endothelium of blood vessels, which may occur during radiotherapy, is discussed as a potential precursor to the development of cardiovascular disease. We thus chose human umbilical vein endothelial cells as a model system to examine the effect of low- and high-linear energy transfer (LET) radiation. Cells were exposed to 250 kV X-rays or carbon ions (C-ions) with the energies of either 9.8 MeV/u (LET = 170 keV/μm) or 91 MeV/u (LET = 28 keV/μm). Subculture of cells was performed regularly up to 46 days (~22 population doublings) post-irradiation. Immediately after exposure, cells were seeded for the colony forming assay. Additionally, at regular intervals, mitochondrial membrane potential (MMP) (JC-1 staining) and cellular senescence (senescence-associated β-galactosidase staining) were assessed. Cytogenetic damage was investigated by the micronucleus assay and the high-resolution multiplex fluorescence in situ hybridization (mFISH) technique. Analysis of radiation-induced damage shortly after exposure showed that C-ions are more effective than X-rays with respect to cell inactivation or the induction of cytogenetic damage (micronucleus assay) as observed in other cell systems. For 9.8 and 91 MeV/u C-ions, relative biological effectiveness values of 2.4 and 1.5 were obtained for cell inactivation. At the subsequent time points, the number of micronucleated cells decreased to the control level. Analysis of chromosomal damage by mFISH technique revealed aberrations frequently involving chromosome 13 irrespective of dose or radiation quality. Disruption of the MMP was seen only a few days after exposure to X-rays or C-ions. Cellular senescence was not altered by radiation at any time point investigated. Altogether, our data indicate that shortly after exposure C-ions were more effective in damaging endothelial cells than X-rays. However, late damage to endothelial cells was not found for the applied conditions and endpoints.

Human Palatal Neoplasm - A Cytopathological Approach

Background: Palatal neoplasm cases in human are very rare. More than 90% neoplasms of the palate is squamous cell type. Not only cytological pleomorphism but also nuclear anomalies are reported to be observed in oral squamous cell carcinoma (OSCC). But, site specific cytological pleomorphisms are not reported so far. Therefore, an attempt has undertaken to investigate the cytopathology of human palatal neoplasms, pattern of cervical lymph node (CLN) metastasis and to analyse the probable etiological risk factors associated with it in the present study. Methodology: In a hospital-based study, out of 136 oral cases, 9 palatal cases (6 male and 3 female) registered during May 2007 to May 2009 were included in this study. Detail case-history including the nature and types of addiction of each individual was recorded prior to the collection of samples. Two scraped exfoliated cytosmears were collected from the affected site on the pre cleaned-coded glass-slides. Collected cytosmears were immediately fixed in 1:3 aceto-alcohol (1 part of glacial acetic acid and 3 parts of ethyl alcohol). One set of smears was stained with Papanicolaou’s stain and the other was counter-stained with Giemsa’s stain for cytopathological analysis. American Joint Committee for Cancer Staging and End-Results Reporting (AJC) formulated TNM (Tumor-Node-Metastasis) System for staging of OSCC was followed. Result: Cytopathologically, a number of pleomorphic cytological atypias, such as, keratinized spindle cell (KSC), keratinized tadpole cell (KTC), keratinized strap (Antischkow) cell (KSC-A), large and small keratinized fiber cells (KFC), large and small keratinized round cells (KRC), micronucleated cell (MNC), plump keratinized squamous cell (PKSC) and non-keratinized malignant squamous cell (NMSC) were observed in such palatal neoplasms. Except NMSCs, all other cells were keratinized. Out of these, PKSC and MNC were well differentiated; KSC, KTC, KFC, KRC and KSCA were moderately differentiated and NMSC was poorly differentiated. Interestingly, in addition to the micronucleated cell (MNC), KSC-A was found to be a modal cytological atypia irrespective of age, site, sex and degree of pathogenicity which may be attributed to the addiction of tobacco in general and smoking in particular. Conclusion: Chewing and smoking of various form of tobacco and drinking of alcohol contribute a lot to the genesis of cytological pleomorphism in human palatal neoplasm. Typically atypical cells like KSC, KTC, KSC-A, KFC, KRC, PKSC, MNC and NMSCs although found less in number, modal occurrence of KSC-A along with the MNC in all the palatal neoplasm may be directly correlated with any form of tobacco smoking.

Buccal Micronuclei Assay as a Tool For Biomonitoring DNA Damage in Oral Lichen Planus.

The malignant transformation rate of Oral Lichen Planus (OLP) is between 0% and 5.8%. Oral lesions of lichen planus clinically presents itself multifocally, simulating the process of field cancerization in high risk malignancies. The Buccal MicroNucleus Cytome Assay (BMN Assay) provides a platform to identify the high risk individuals by evaluating the markers of nuclear damage at an earliest micro invasive phase. To evaluate DNA damage in exfoliated buccal mucosal cells in individuals with oral lichen planus lesions and thereby to delineate the high risk group. Buccal smears from 22 OLP and 10 control samples were stained in modified Feulgen-Rossenback reaction for micronuclei assay. Cytological evaluation of number of MicroNucleated cells (CMN), Total Number of Micronuclei (TMN) in micronucleated cells was done in both groups. Frequency of micronucleated cells (CMN) when compared among the study and control group, a mean value of 4.27 ± 1.80 and 0.90 ± 0.88 were obtained respectively. On comparing the total number of micronuclei in the micronucleated cells (TMN) between the study and control groups, a mean value of 5.38 ± 2.42 and 1.5 ± 0.88 were obtained respectively. There was a significant increase in the frequency of micronuclei and the micronucleated cells in the oral lichen planus as compared to normal individuals.

Biomonitoring with Micronuclei Test in Buccal Cells of Female Farmers and Children Exposed to Pesticides of Maneadero Agricultural Valley, Baja California, Mexico.

Feminization of the agricultural labor is common in Mexico; these women and their families are vulnerable to several health risks including genotoxicity. Previous papers have presented contradictory information with respect to indirect exposure to pesticides and DNA damage. We aimed to evaluate the genotoxic effect in buccal mucosa from female farmers and children, working in the agricultural valley of Maneadero, Baja California. Frequencies of micronucleated cells (MNc) and nuclear abnormalities (NA) in 2000 cells were obtained from the buccal mucosa of the study population (n = 144), divided in four groups: (1) farmers (n = 37), (2) unexposed (n = 35), (3) farmers' children (n = 34), and (4) unexposed children (n = 38). We compared frequencies of MNc and NA and fitted generalized linear models to investigate the interaction between these variables and exposition to pesticides. Differences were found between farmers and unexposed women in MNc (p < 0.0001), CC (p = 0.3376), and PN (p < 0.0001). With respect to exposed children, we found higher significant frequencies in MNc (p < 0.0001), LN (p < 0.0001), CC (p < 0.0001), and PN (p < 0.004) when compared to unexposed children. Therefore working as a farmer is a risk for genotoxic damage; more importantly indirectly exposed children were found to have genotoxic damage, which is of concern, since it could aid in future disturbances of their health.

Exfoliative Cytopathology of Human Lip Neoplasm

Objective: The objective of the present study is to investigate the cytopathology of human lip neoplasms, pattern of cervical lymph node (CLN) metastasis and to analyse the probable etiological risk factors associated with it. Methodology: In this hospital based case-control study, 22 subjects (11 cases of lip neoplasm and 11 healthy individuals as Control group) were included in this study. Scraped exfoliated cytosmears were collected from the affected site of the lip and smearing was done in the precleaned- coded glass-slides. Two such slides were prepared from each subject. The cytosmears were immediately fixed in aceto-alcohol (1part of glacial acetic acid:3 part of absolute ethyl alcohol) fixative. One set of the slide was stained with Papanicolaou’s stain and the other set wascounter-stained with Giemsa’s Solution for cytopathological analysis.The TNM (Tumor- Node-Metastasis) System for staging of lip cancer formulated by American Joint Committee for Cancer (AJCC) Staging and End Results Reporting-2010 was followed. Results: Lower lip is found to be the most common site in the genesis of lip neoplasm. Pleomorphic cytological atypias such as Micronucleated cell (MNC), Plump keratinized squamous cell (PKSC), Keratinized spindle cell (KSC), Keratinized spindle cell (KSC), Keratinized tadpole cell (KTC), Keratinized strap (Anitschkow) cell (KSC-A), Keratinized fiber cell (KFC), Keratinized round cell (KRC) and Non-keratinized malignant squamous cell (NMSC) with drastic modification were frequently observed in lip neoplasm. Unpredictable metastasis is a common feature in lip neoplasm cases. Conclusion: Excluding PKSC and MNC, presence of any other atypias in the cytosmearoflip neoplasm indicates the state of malignancy. Thus, exfoliative cytopathology has a potentiality in early detection of lip carcinoma in particular and oral cancer in general.