Induction of Chromosomal Aberrations by Propoxur in Mouse Bone Marrow Cells

Propoxur is a widely used dithiocarbamate insecticide. In this study, the clastogenic effect of propoxur has been evaluated using chromosomal aberration assay in mouse bone marrow cells. Single i. p. administration of propoxur, at 25 mg/kg b.wt., a maximum tolerated dose (MTD) and 12 .5mg/kg b.wt (50% of MTD) have significantly induced different types of aberrations after 24 h of treatment. The aberrations were dose and time dependent and reached a maximum after 24 h of exposure. The sresult suggest a genotoxic potential of propoxur.

Evaluation of the hepatoprotective and antioxidant effects of Tegillarca granosa flesh body extract against potassium bromide toxicity via targeting the histomorphometry,chromosomal and expressions of TGF-β1,VEGF and COX-2 genes in rats

The hepatotoxic effect of potassium bromide(KBr)on rat liver tissues were determined,as well as the potential protective effect of Tegillaraca granosa(T.granosa)flesh body extract.Twenty adult male albino rats were equally distributed into four groups;Group(I)treated with physiological saline(control group),Group(II)was orally gavaged by 200 mg/kg of T.granosa body extract day after day,Group(III)was intoxicated by KBr(150 mg/kg bwt day after day orally)and finally,Group(IV)was given a combination of T.granosa flesh body extract plus KBr with similar doses in the second and third groups.At the end of one month,blood,liver tissue and bone marrow samples were collected to be used for the required laboratory examinations.In response to KBr toxicity,there was a significant increase in serum antioxidant biomarkers,which was accompanied by a significant change in hepatocyte ultrastructure and a significant change in carbohydrate and protein levels within the liver organ.In addition,KBr intoxication resulted in a substantial increase in the incidence of chromosomal aberrations such as holes,splits,deletions,fragments,ploidy,and ring chromosomes,as well as significant upregulation of TGF-1,VEGF,and COX-2 gene expression.The hepatotoxic effect of KBr was counteracted by treatment with T.granosa flesh body extract.T.granosa flesh body extract has a curative antioxidant and numerous protective effects against KBr hepatotoxicity.

Acute toxicity and chromosomal aberration toxicity of insect wax and its policosanol

An acute toxicity test in Sprague Dawley(SD)rats and a chromosomal aberration toxicity test in Chinese hamster lung(CHL)fibroblasts were conducted to promote the application of insect wax and its policosanol.Results of oral acute toxicity test showed that the LD;values of insect wax and its policosanol were higher than 5000 mg/kg.The rats showed no obvious toxic symptoms and survived,suggesting that these substances were not toxic.The chromosomal aberration rates of the CHL cells incubated with insect wax for 3-6 h under metabolic activation conditions and for 3-6 h and 24 h under non-metabolic activation conditions were lower than 5%.No significant difference was found compared with the negative control group.Also,no significant difference in chromosomal aberration rates was found between each dose group of policosanol under metabolic or non-metabolic activation conditions and the negative control group.Therefore,the chromosomal aberration rates of CHL cells treated with insect wax and its policosanol were negative regardless of metabolic activation conditions.These results indicate the absence of acute toxicity and potential chromosomal aberration in the tested dose range of insect wax and its policosanol.

The Level and Distribution of Chromosomal Aberration of Tomato Seeds at Different Penetration Depths of Carbon Ions

The relationship between the penetration depth and the level and distribution of chromosomal aberration of the root tip cells were investigated by exposure of the superposed tomato seeds to 80 MeV/u carbon ions. The results showed that on the entrance of the beam the chromosomal aberration level was low. Damage such as breaks aud gaps were dominant. At the Bragg peak~ the chromosomal aberration level was high. The yields of dicentrics~ rings and disintegrated small chromosomes increased but the yields of breaks and gaps decreased. These results are consistent with the distribution of tile physical depth dose profile of carbon ions. It is effective to deposit the Bragg peak on the seeds to induce hereditary aberration in the mutation breeding with heavy ions.

Effects of CO_2 laser on chromosomal aberration in the root tip cells of rice

Seeds of japonica rice cv.Zhenuo 2 with twodifferent physiological states(dry seeds withwater content 13% and wet seeds soaked in thewater for 36 h)were irradiated by COlaser infour different power-densities and durations re-spectively.The treatment irradiated with 200GY ofCo γ-rays was considered as control.The flesh root tips were cut and fixed inCarnoy’s fluid for cytological examination.

Genotoxic Effects of PAH Containing Sludge Extracts in Chinese Hamster Ovary Cell Cultures

Objective Many studies have been conducted in order to evaluate the genotoxicity of chemicals and waste materials, which utilized in vivo test protocols. The use of animals for routine toxicity testing is now questioned by a growing segment of society. Methods Keeping the above fact in mind, we have conducted in the present study the genotoxicity evaluation of oily sludge samples generated from a petroleum refinery and petrochemical industry and ETP sludge from petroleum refinery using DNA damage, chromosomal aberration, p53 protein induction and apoptosis in short term in vitro mammalian Chinese Hamster Ovary cell cultures. Results It is evident from the results that the oily sludge compounds derived from petroleum refinery and petrochemical industry could cause DNA damage, chromosomal aberration, p53 protein accumulation and apoptotic cell death on exposure to oily sludge extracts in the presence of metabolic activation system (S-9 mix), however, ETP sludge extract could not cause significant genotoxicity in comparison to oily sludge extract and negative control. Conclusion The effect may be attributed to polycyclic aromatic hydrocarbons present in the samples as evidenced from GC-MS.

In vitro Antigenotoxicity of Ulva rigida C. Agardh (Chlorophyceae) Extract against Induction of Chromosome Aberration, Sister Chromatid Exchange and Micronuclei by Mutagenic Agent MMC

Objective To determine the in vitro possible clastogenic and cytotoxic activities of Ulva rigida crude extracts （URE）, and identify their antigenotoxic and protective effects on chemotherapeutic agent mitomycine-C （MMC）. Methods Anti-clastogenic and anti-genotoxic activities of Ulva rigida crude extracts （URE） were studied using chromosome aberration （CA）, sister chromatid exchange （SCE）, and micronuclei （MN） tests in human lymphocytes cultured in vitro. Results The chromosome aberration, sister chromatid exchange or micronuclei tests showed that URE at concentrations of 10, 20, and 40 lag/mL had no clastogenic activity in human lymphocyte cell culture. Three doses of URE significantly decreased the number of chromosomal aberrations and the frequencies of SCE and MN when compared with the culture treated with MMC （P〈0.0001）. Conclusion Although URE itself is not a clastogenic or cytotoxic substance, it possesses strong antigenotoxic, anti-clastogenic, and protective effects on MMC in vitro.

Evaluation of Cytogenetic Effects of Isoproturon on the Root Meristem Cells of Allium sativum

Objective\ To validate the use of Allium sativum as a reliable test model for genotoxicity, isoproturon, a substituted phenylalkylurea herbicide, was evaluated on the root meristem cells by this plant system. Method\ Test concentrations were selected by determining EC50 and root tips were exposed to various concentrations for 6 or 24 hr. EC50 concentration was calculated to be 70.8 ppm for the root growth. In addition to root growth retardation exposure to isoproturon induced morpholoogical changes like discolouration and stiffness of roots. Results\ Exposure to various experimental concentrations of isoproturon (35-280 ppm), including EC50, significantly and dose-dependently inhibited the mitotic index and induced chromosome breaks/mitotic aberrations at 6 or 24 hr. Conclusion\ The frequency of aberrations was found to be decreased in a dose dependant manner at 24 or 48 hr post exposure, however in comparison of control cells the frequency of aberrations was considerably high which indicates genotoxicity potentials of isoproturon. Further, present study also suggests that Allium sativum is a sensitive, efficient, and reliable test system for measuring the genotoxicity potential of environmental chemicals.

ANEUPLOIDY INDUCTION BY THE WATER EXTRACT OF TRIPTERYGIUM HYPOGLAUCUM(LEVEL) HUTCH IN MOUSE BONE MARROW CELLS

The aneuploldy inducing activity of a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH) were investigated by means of thhree cytogenetic end points, namely C-mitotic (CM) effects, micronucleus (MN) and parallel chromosome structural aberration (CA) analyses in vivo. The experiments were performed on mouse bone marrow cells. THH showed similar gentoxic effects to colchicine (COL) in CM, MN and CA analyses; positive CM effects were observed accompanied with increases of mitotic index and frequencies of C-mltotic cells as well as decreased frequencies of anaphase in all of the THH-treated groups. The compound showed a positive MN response in bone marrow polychromatic erythrocytes but was negative in CA analyses. The preliminary results suggested that THH is an aneuploldy inducer in mouse bone marrow cells under present experiment conditions.

Genotoxicity of Pesticide Waste Contaminated Soil and Its Leachate

Objective Improper land disposal of hazardous waste can result in leaching of hazardous constituents which may contaminate ground and surface water leading to adverse impact on human health and environment consequences. The present study utilized mammalian cell culture for the genotoxicity assessment of waste and its leachate. Methods Genotoxic potential and chemical analysis of pesticide derived tarry waste contaminated soil extract and its leachate was assessed using in vitro human lymphocyte cultures and GC-MS. Results The investigation revealed that the soil extract could cause significant to highly significant genotoxicity in the form of DNA strand break at 25 mL (P<0.01), 50 mL, 100 mL and 200 mL (P<0.001) and chromosomal aberration at 25 mL (P<0.01) and 50 mL and 100 mL (P<0.001). The leachate could cause significant DNA strand break and chromosomal aberration only at 100 mL and 200 mL (P<0.01) dose levels. Conclusion The genotoxicity observed is attributed to carbaril and tetra methyl naphthyl carbamate, the major ingredients of the extracts, as revealed by GC-MS.

Combination of Cytogenetic Analysis and Molecular Screening in Patients with de novo Acute Myeloid Leukemia

Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations in leukemia,we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction(RT-PCR) and fluorescence in situ hybridization(FISH),and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML.Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients,and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH.The PCR products in some suspected cases were tested by two-directional sequencing.The results showed that except unqualified samples,fusion genes were detected by multiplex RT-PCR in 211 of 474 patients(44.51%),including AML1-ETO,CBFβ-MYH11,PML-RARα,PLZF-RARα,NPM-RARα,MLL rearrangements,BCR-ABL,DEK-CAN,SET-CAN,TEL-PDGFR,TLS-ERG,AML1-MDS1(EVI-1).In 373 patients,who took both multiplex RT-PCR and karyotype analysis,the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373(42.89%) and 179/373(47.98%) respectively,and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373(57.90%).The PCR results from 11 cases 'normal' in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing.It is concluded that karyotype studies remain the cornerstone for genetic testing;conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML,especially for the cryptic or submicroscopic aberrations.Once a genetic marker has been identified by combined analysis,it could be used to monitor residual disease during/after chemotherapy,by quantitative RT-PCR and/or FISH.

Mathematical Modeling of Carcinogenesis Based on Chromosome Aberration Data

Objective： The progression of human cancer is characterized by the accumulation of genetic instability. An increasing number of experimental genetic molecular techniques have been used to detect chromosome aberrations. Previous studies on chromosome abnormalities often focused on identifying the frequent loci of chromosome alterations, but rarely addressed the issue of interrelationship of chromosomal abnormalities. In the last few years, several mathematical models have been employed to construct models of carcinogenesis, in an attempt to identify the time order and cause-and-effect relationship of chromosome aberrations. The principles and applications of these models are reviewed and compared in this paper. Mathematical modeling of carcinogenesis can contribute to our understanding of the molecular genetics of tumor development, and identification of cancer related genes, thus leading to improved clinical practice of cancer.

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.

Anticlastogenic Effect of Redistilled Cow's Urine Distillate in Human Peripheral Lymphocytes Challenged With Manganese Dioxide and Hexavalent Chromium

Objective To study the anticlastogenic effect of redistilled cow＇s urine distillate （RCUD） in human peripheral lymphocytes （HLC） challenged with manganese dioxide and hexavalent chromium. Methods The anticlastogenic activity of redistilled cow＇s urine distillate was studied in human polymorphonuclear leukocytes （HPNLs） and human peripheral lymphocytes in vitro challenged with manganese dioxide and hexavalent chromium as established genotoxicants and clastogens which could cause induction of DNA strand break, chromosomal aberration and micronucleus. Three different levels of RCUD： 1 μL/mL, 50 μL/mL and 100 μL/mL, were used in the study. Results Manganese dioxide and hexavalent chromium caused statistically significant DNA strand break, chromosomal aberration and micronucleus formation, which could be protected by redisfilled cow＇s urine distillate. Conclusion The redistilled cow＇s urine distillate posseses strong anfigenotoxic and antielastogenic properties against HPNLs and HLC treated with Cr^＋6 and MnO2. This property is mainly due to the antioxidants present in RCUD.

Modulatory Effect of Distillate of Ocimum sanctum Leaf Extract (Tulsi) on Human Lymphocytes Against Genotoxicants

Objective To study the modulatory effect of distillate of Ocimum sanctum （traditionally known as Tulsi） leaf extract （DTLE） on genotoxicants. Methods In the present investigation, we studied the antigenotoxic and anticlastogenic effect of distillate of Tulsi leaf extract on （i） human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C （MMC） and hexavalent chromium （Cr^＋6） and （ii） human peripheral lymphocytes （in vitro） with or without metabolic activation against mitomycin C （MMC）, hexavalent chromium （Cr^＋6） and B[a]P by evaluating chromosomal aberration （CA） and micronucleus assay （MN）. Three different doses of DTLE, 50 μL/mL, 100 μL/mL, and 200 μL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break, chromosomal aberration and micronucleus emergence. The following positive controls were used for inducing genotoxicity and clastogenicity MMC （0.29 μmol/L） for DNA strand break, chromosomal aberration and 0.51 μmol/L for micronucleus assay; Potassium dichromate （Cr^＋6） 600 μmol/L for DNA strand break and 5 μmol/L for chromosomal aberration and micronucleus assay; Benzo[a]pyrene （30 μmol/L） for chromosomal aberration and 40 μmol/L for micronucleus assay. The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS. Results Mitomycin C （MMC） and hexavalent chromium （Cr^＋6） induced statistically significant DNA strand break of respectively 69% and 71% （P〈0.001） as revealed by fluorometric analysis of DNA unwinding. Furthermore, the damage could be protected with DTLE （50 μL/mL, 100 μL/mL, and 200 μL/mL） on simultaneous treatment. Chromosomal aberration and micronucleus formation induced by MMC, Cr^＋6 and B[a]P were significantly protected （P〈0.001） by DTLE with and without metabolic activation. Conclusion Distillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol, luteolin and apigenin as identified by LC-MS. These active ingredients may have the protective effect against genotoxicants.

Cytogenetic and andrological status and ICSI-results in couples with severe male factor infertility

Aim: To pursue whether cytogenetic aberrations correlate with specific spermatological or hormonal abnormalities.Methods: 305 infertile couples were investigated. All male partners were referred to a complete andrological work-up with physical examination, determination of hormones, HIV testing and semen analysis. Cytogenetic analysis wascarded out in both partners by means of standard techniques using cultured lymphocytes from peripheral blood. Re-suits: Among the 305 couples, 10 men (3.2%) and 10 women (3.2%) showed constitutional chromosomal aber-rations, including reciprocal translocations (n = 7), Robertsonian translocations (n = 3), inversions (n = 3), otherstructural aberrations (n = 4) and sex chromosome aberrations (n = 3). In addition to the impaired sperm count inmost of the patients, a tendency to an increased proportion of spermatozoa with acrosome defect was observed. Con-clusion: Chromosomal aberrations may contribute to the low fertilization and pregnancy rates in the infertile couples.(Asian J Androl 2000 Dec; 2: 293-296)

In vitro Safety Evaluation and Anticlastogenic Effect of BacoMind^(TM) on Human Lymphocytes

Objective BacoMind^TM （BM） is a standardized extract of Bacopa monnieri, which belongs to the family Scrophulariaceae and is a creeping annual plant found throughout the Indian subcontinent. It has been used by Ayurvedic medicinal practitioners in India for almost 3000 years and is classified as a medharasayana, a substance which improves memory and intellect. With the widespread traditional use as well as scientific validation of Bacopa monnieri for nootropic activity, a bioactive-rich unique phytochemical composition-BacoMind^TM was developed from B. monnieri for use as a cognition and memory enhancing agent. The present study aimed to investigate the in vitro toxicity of this formulation of BacoMind^TM on human lymphocytes and to rule out its possible contribution to mutagenicity. Methods In the present investigation the active ingredients present in BM were identified and quantified by high performance liquid chromatography （HPLC） and high performance thin-layer chromatography （HPTLC）. Antioxidant and anticlastogenic properties of BM were studied in vitro with and without metabolic activation. Doses of BM were chosen on the basis of mitotic index （MI） and cytokinesis-block proliferation index （CBPI）. Clastogenicity assays were performed at 31.2 μg/mL, 62.5 μg/mL, and 125 μg/mL, while the Salmonella reverse mutation assay （Ames test） was performed at doses of 61.72, 185.18, 555.55, 1666.67, and 5000.00 μg/plate. Results HPLC and HPTLC analysis of BM revealed the presence of bacoside A3. bacopaside Ⅰ, bacopaside Ⅱ, jujubogenin isomer of bacopasaponin C, bacosine, luteolin, apigenin, bacosine, and β-sitosterol D glucoside. BM demonstrated significant antioxidant activity. The number of chromosomal aberrations and the frequency of micronuclei induced by BM were not statistically significant up to a dose of 62.5 μg/mL. A subsequent dose of 125 μg/mL prior to metabolic activation induced mild clastogenicity, but it was found to be biologically insignificant as this effect was not seen post metabolic activation. BM also demonstrated a dose-dependent protection against the clastogens used in this study using the above tests for clastogenicity. Maximum protection was observed in presence of metabolic activation. Moreover, BM demonstrated no mutagenic effect on the tested strains, as observed in the Ames test. Conclusion BM protected human lymphocytes against various clastogens. BM also exhibited high antioxidant activity which might be responsible for the observed protective effects against the clastogens since the used clastogens are known to induce their clastogenic effects via production of oxidative radicals.

Evaluation of the in vitro and in vivo Genotoxicity of Almond Skins

Objective It aims to study potential genotoxicity of almond skins.Methods A bacterial reverse mutation assay was performed on S.typhimurium strains TA97,TA98,TA100,TA102,and TA1535 in the absence or presence of S-9 mixture at a dose range of 312.5 to 5 000 μg/plate.A micronucleus test and a mammalian bone marrow chromosome aberration tests were performed in Swiss Albino （CD-1） mice at doses of 625,1 250,and 2 500 mg/kg bw used.Results Almond skins exerted no mutagenic activity in various bacterial strains of Salmonella typhimurium in either the absence or the presence of metabolic activation at all doses tested.Various doses of almond skins did not affect the proportions of immature to total erythrocytes,the number of micronuclei in the immature erythrocytes,or the number of structural and numerical chromosomal aberrations of Swiss albino mice.Conclusion Almond skins are not genotoxic under the conditions of the in vitro bacterial reverse mutation assay and two in vivo tests-micronucleus test and mammalian bone marrow chromosome aberration test,which supports the safety of almond skins for dietary consumption.

Effects of Zn^（2＋） on root growth，cell division，and nucleoli of Allium cepa L．

The effects of different concentrations of Zn￣(2+)ion on root growth,cell division,and nucleoli of Allium cepa were studied. The test Zn￣(2+) ion concentration was made up from zinc sulphate (ZnSO4. 7H2O) ranging from 10￣(-7) to 10￣(-2) mol/L. The solutions were prepared in tap water (pH =6. 5).The results indicated that Zn￣(2+) could obviously inhibit root growth at concentrations from 10￣(-4)to 10￣(-2) mol/L.Roots treated with zinc sulphate showed the presence of c-mitosis, anaphase bridges,including sticky and fluidized bridges (at 10￣(-3) to 10￣(-2) mol/L) , chromosome stickiness, irregularly shaped nuclei, broken nuclei and micronuclei. A toxicity effect was also observed on the nucleoli using silver staining technique after 48h of treatment with 10￣(-4)to 10￣(-2) mol/L Zn￣(2+), e. g,the nucleolar particulate material scattered around the nucleoli in the nucleus of root tip cells.

A study on the genetic noxiousness of trinitrotoluene

In this paper, the mutagencity of TNT was studied for mice and fruit flies. It was indicated that TNT had a great deal of mutagenic effect on the somatic ce1ls and sexual cells of the animals used as the subjects of the experiment, and calculated that TNT would have a genetic harmfulness to human beings.