Antimutagenic potential of curcumin on chromosomal aberrations in Allium cepa

Turmeric has long been used as a spice and food colouring agent in Asia. In the present investigation, the antimutagenic potential of curcumin was evaluated in Allium cepa root meristem cells. So far there is no report on the biological properties of curcumin in plant test systems. The root tip cells were treated with sodium azide at 200 and 300 μg/ml for 3 h and curcumin was given at 5, 10 and 20 μg/ml for 16 h, prior to sodium azide treatment. The tips were squashed after colchicine treatment and the cells were analyzed for chromosome aberration and mitotic index. Curcumin induces chromosomal aberration in Allium cepa root tip cells in an insignificant manner, when compared with untreated control. Sodium azide alone induces chromosomal aberrations significantly with increasing concentrations. The total number of aberrations was significantly reduced in root tip cells pretreated with curcumin. The study reveals that curcumin has antimutagenic potential against sodium azide induced chromosomal aberrations in Allium cepa root meristem cells. In addition, it showed mild cytotoxicity by reducing the percentage of mitotic index in all curcumin treated groups, but the mechanism of action remains unknown. The antimutagenic potential of curcumin is effective at 5 μg/ml in Allium cepa root meristem cells.

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.

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.

Identification of a high frequency of chromosomal rearrangements in the centromeric regions of prostate cancer patients

The aim of the present investigation was to study the major chromosomal aberrations （CA） like deletion, translocation, inversion and mosaic in prostate cancer patients of Tamilnadu, Southern India. Totally 45 blood samples were collected from various hospitals in Tamilnadu, Southern India. Equal numbers of normal healthy subjects were chosen after signing a consent form. Volunteers provided blood samples （5 ml） to establish leukocyte cultures. Cytogenetic studies were performed by using Giemsaanding technique and finally the results were ensured by spectral karyotyping （SKY） technique. In the present investigation, major CA like deletion, translocation, inversion and mosaic were identifed in experimental subjects. Results showed frequent CA in chromosomes 1, 3, 5, 6, 7, 9, 13, 16, 18 and X. In comparison with experimental subjects, the control subjects exhibited very low levels of major CA （P〈0.05）. In the present study, the high frequency of centromeric rearrangements indicates a potential role for mitotic irregularities associated with the centromere in prostate cancer tumorigenesis. Identification of chromosome alterations may be helpful in understanding the molecular basis of the disease in better manner.

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.

Studies on Ultra-dry Storage and Genetic Stability of Vegetable Seeds

[Objective] To study the long-term ultra-dry storage method and genetic stability of vegetable seeds.[Method] Seeds of Lycopersicum esculentum,Raphanus satuvus and Apium graveolen.were chosen as material.The changes of seed vigor,viability and genetic stability after ultra-storage were discussed by studying the seed potentiality,shoot length,germination percentage and the chromosome aberration rate of root tip cells.[Results] Maintaining the low moisture content,different vegetable species had different storage effects of the long-term storage seeds under normal temperature.The Lycopersicum esculentum and Raphanus satuvus seeds were more suitable to ultra-dry storage at normal temperature,and could keep good genetic stability,while the seeds of Apium graveolen had bad performance.[Conclusion] This study established the foundation of studying ultra-dry storage of vegetable seeds.

Copy number changes of target genes in chromosome 3q25.3-qter of esophageal squamous cell carcinoma: TP63is amplified in early carcinogenesis but down-regulated as disease progressed

AIM: By using comparative genomic hybridization, gain of 3q was found in 45-86% cases of esophageal squamous cell carcinoma (EC-SCC). Chromosome 3q25.3-qter is the minimal common region with several oncogenes found within this region. However, amplification patterns of these genes in EC-SCC have never been reported. The possible association of copy number changes of these genes with pathologic characteristics is still not clear. METHODS: Real-time quantitative PCR (Q-PCR) was performed to analyze the copy number changes of 13 candidate genes within this region in 60 primary tumors of EC-SCC, and possible association of copy number changes with pathologic characteristics was analyzed by statistics. Immunohistochemistry (IHC) study was also performed on another set of 111 primary tumors of EC-SCC to verify the association between TP63 expression change and lymph node metastasis status. RESULTS: The average copy numbers (±SE) per haploid genome of individual genes in 60 samples were (from centromere to telomere): SSR3: 4.19 (±0.69); CCNL1: 5.24 (±0.67); SMC4L1: 2.01 (±0.16); EVI1: 2.02 (±0.12); hTERC. 5.28 (±0.54); SKIL 2.71 (±0.14); EIF5A2. 1.95 (±0.12); ECT2: 9.18 (±1.68); PIK3CA: 8.13 (±1.17); EIF4G1: 1.07 (±0.05); 557: 3.07 (±0.25); TP63: 2.51 (±0.22); TFRC. 2.42 (±0.19). Four clusters of amplification were found: SSR3 and CCLN1 at 3q25.31; hTERC and SKIL at 3q26.2; ECT2 and PIK3CA at 3q26.31-q26.32; and 55T, TP63 and TFRC at 3q27.3-q29. Patients with lymph node metastasis had significantly lower copy number of TP63 in the primary tumor than those without lymph node metastasis. IHC study on tissue arrays also showed that patients with lymph node metastasis have significantly lower TP63 staining score in the primary tumor than those without lymph node metastasis. CONCLUSION: This study showed that different amplification patterns were seen among different genes within 3q25.3-qter in EC-SCC, and several novel candidate oncogenes (SSR3, SMC4L1, ECT2, and SST) were identified. TP63 is amplified in early stage of EC-SCC carcinogenesis but down-regulated in advanced stage of disease.

Mutagenic effects of chromium trioxide on root tip cells of Vicia faba

In this study on the mutagenic effects of different concentrations of chromium trioxide (CrO3) on Vicia faba root tip, micronucleus assay and chromosome aberration assay were used to determine the mitotic indexes, micronucleus rate and chromosome aberration rate of Vicia faba root tip cells. The results showed that the effects of CrO3 concentration on the mitotic indexes were complicated. CrO3 increases the micronucleus rate of Vicia faba root tip cells. It was found that within certain range of CrO3 concentration the micronucleus rate increased systematically with increased concentration of CrO3, but that the micronucleus rate decreased at higher level of CrO3 and that CrO3 also caused various types of chromosome aberration at a rate which increased systematically with increased concentration of CrO3. We concluded that CrO3 has significant mutagenic effect on Vicia faba root tip cells.

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.

Cytogenetic Effects of Pulsing Electromagnetic Field on Domestic Pig Lymphocytes in Vitro

The effects of pulsing electromagnetic fields(PEMFs)on cells are very important subjects in the field of bioelectromagnetics.In this experiment,the cytogenetic effects of PEMF on domestic pig lymphocytes were tested in vitro.Pig lymphocytes in RPMI 1640 medium were exposed to PEMFs of 100 kHz and 200 kHz for 12,24 and 48 hours.Chromosomal aberrations(aneuploidy,breaks,gaps,et al)were significantly increased in exposed cultures,and of these aberrations,56%chromosomal or chromatid breaks and 42%gaps induced by PEMFs were the points of pig chromosomal fragile sites.The baseline frequency of sister chromatid exchange(SCE)increased after exposing lymphocytes continuously to PEMFs of 100 kHz and 200 kHz for 48 hours.These results suggested that the exposure to PEMFs might induce a type of DNA lesion and chromosomal aberrations.

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.