{"paper_id":"113b4aa4-c009-4614-9e8d-59a7d8351544","body_text":"Genotoxicity, DNA damage and sperm defects induced by vinblastine | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Genotoxicity, DNA damage and sperm defects induced by vinblastine Maha A. Fahmy, Entesar Hassan, Ayman A. Farghaly, Zeinab M. Hassan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1943872/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Background Genotoxicity studies of chemotherapeutic drugs is of special need. Secondary tumors may develop many years after treatment as a result of chemo genotoxicity. The effect of chemo on meiotic chromosomes and sperm defects is another complication associated with chemo treatment. In this study the genotoxicity of vinblastine (VB) was estimated in both somatic and germ cells. Materials 85 mice were taken. 4 single doses of VB at 3, 4.5, 6 and 10 mg/kg and 3 successive doses at 3, 4.5 and 6 mg/kg were taken for estimation of chromosomal aberrations (CAs). 4 single doses of VB were involved in estimating the DNA fragmentation, and comet assay. Samples were taken 24 h after the last treatment. For sperm abnormalities mice were injected with 3 successive doses of VB at 3, 4.5, and 6 mg/kg and samples were taken 35 days after the 1st injection. Results The results demonstrated a significant frequency of DNA fragmentation in spleen cells and in the percentage of CAs in bone marrow. Numerical and structural aberrations were recorded with a pronounced number of polyploidy metaphases. VB also induced a significant percentage of CAs in spermatocytes in the form of univalent. Sperm defects in the form of coiled tail, absence of acrosome and shapeless head and a significant DNA damage in the testes were recorded. Conclusion VB is genotoxic in somatic and germ cells. Sperm defects induced by VB are of serious concern to future generations and may affect the fertility of cancer survivors. Vinblastine Mitotic inhibitor Chromosomal aberrations DNA damage Sperm abnormalities Figures Figure 1 Figure 2 Figure 3 Introduction Vinblastine (VB) is a very important clinic alkaloid extracted naturally from a periwinkle plant which is a flowering herb from Catharanthus roseus [ 1 ]. It is a chemotherapeutic medication used separately or in combination with other cytotoxic drugs (bleomycin, doxorubicin, and methotrexate…) for the treatment of a wide range of human neoplasms e.g. Hodgkin and non-Hodgkin’s lymphoma, T-cell lymphoma, testicular and germ cell tumors, bladder, lung (Non-small cell lung cancer), head and neck cancers. It is also used to treat melanoma, soft tissue sarcoma, Kaposi’s sarcoma, gestational trophoblastic and some blood disorders ( e.g. histiocytosis X) [ 2 , 3 ]. In some cases it exerts an immunosuppressive effects. VB is one of the World Health Organization List of Essential Medications. It is a member of the Vinca alkaloid family which are anticancer agents that include the natural product vinblastine, vincristine, and the semi-synthetic product vin-orelbine [ 4 ]. Vinca alkaloids have the potential to induce cytotoxic and genotoxic effects, oxidative DNA damage and micronuclei formation [ 2 , 5 ]. Vinca alkaloids exert their function as mitotic spindle inhibitors [ 6 ].They are antimitotic drugs that inhibit microtubules and are cell cycle phase-specific ´S and M phase poisons´ [ 7 ]. VB is potentially mutagenic and animal studies have demonstrated degenerative changes in germ cells [ 8 ], and reported an inhibition of angiogenic action [ 9 ]. A major problem with cancer chemotherapy is its severe toxic effect on non-target tissues (normal cells), so the cancer patients not only suffer from an immediate side effects of cancer medications but are also subjected to an increased risk of developing secondary tumors later in life [ 10 ]. Recent studies in chemotherapy approaches claim at developing medications that eradicate cancer cells efficiently and selectively. So it is of special need to study the side effects of anticancer agents in order to improve the conventional chemotherapy by protecting healthy tissues. The genotoxic reports concerning VB are controversial, limited and inconclusive. The aim of the present work is to determine the genotoxic effects of VB using different mutagenic biomarkers: chromosomal aberration analysis and DNA damage in both somatic and germ cells, and also assessment of sperm defects induced in male mice. Materials And Methods Chemicals Common trade name of the chemotherapeutic drug is vinblastine sulfate, velban and others. Vinblastine was obtained from Sigma Aldrich (MO,USA). Drug Bank Accession Number: DB00570. All other chemicals were purchased from ADWICC (Cairo, Egypt). Animals Male white Swiss mice ( Mus musculus ), aged 9 to 12 weeks, were used in all experiments. Animals were obtained from a closed random-bred colony at the National Research Centre (Giza, Egypt). Mice used for each experiment were of similar age (±1 week) and weight (± 2 g). Animals were housed in polycarbonate boxes with steel wire tops and bedded with wood shavings. Ambient temperature was controlled at 22°C ± 3°C with a relative humidity of 50% ± 15% and a 12 h light/dark photoperiod. Food and water were provided ad libitum . The experiments were conducted according to the Animal Research Ethical Committee Guidelines of the National Research Centre, Egypt. The Approval Certificate is under number: 19 163. Experimental design In these experiments a total of 85 mice were taken as follows: 40 mice for chromosomal aberrations in bone marrow and mouse spermatocytes, 25 mice for DNA fragmentation and alkaline comet assay and 20 for morphological sperm abnormalities. In each of these testes, mice were subdivided into groups (5 /group). The main groups represented chromosomal aberration analysis: Group I: Negative control, Groups II-V, mice were i.p injected with a single dose of VB (3, 4.5, 6 and 10 mg/kg), Groups VI-VIII in which mice received repeated i.p injections (three successive days) of VB at the dose levels 3, 4.5 and 6 mg/kg. In all experiments samples were taken 24h after the last treatment. Analysis for DNA damage, showed five groups: Animals were treated with a single i.p injection of VB at the dose levels 3, 4. 5, 6 and 10 mg/kg, in addition to the control group. Samples were taken 24h after injection. Analysis of sperm abnormalities showed four groups as follows: Negative control group and three treated groups with VB (3, 4.5 and 6 mg/kg, 3 injections) and samples were taken 35 days after the 1 st injection. Different doses were taken to cover all doses required for different types of human cancerous. Cytogenetic analysis Chromosomal aberration assay in mouse bone marrow and spermatocytes Mitotic and meiotic chromosomes were prepared from bone marrow and testis of the same animal, respectively. Bone marrow chromosomes were prepared according to the technique described by Diab et al. [11]. In brief, mouse bone- marrow cells were collected from both femurs, cells were incubated in hypotonic solution (KCL 0.075 M) for 20 min at 37°C, and then centrifuged. The cell pellets were suspended in a fixative (methanol/glacial acetic acid 3:1). This step was repeated at least twice, then the cells were suspended in a few drops of fixative and spread onto frozen slides, air-dried, stained with 10% Giemsa for 30 min, washed, and air dried again. Spermatocyte chromosomes were prepared from the testes according to the protocol described by Evans et al.[12]with some modifications [13]. Briefly, the testis was removed and squashed into a petri dish containing an isotonic solution of 2.2% trisodium citrate. Then the cell suspension was centrifuged for 5 minutes at 1500 rpm. The cell pellet was incubated in a hypotonic solution of 1.1% trisodium citrate for 20 minutes at 37C◦ followed by centrifugation. The cell pellet was washed twice by a freshly prepared fixative. A few drops of the fixative cell suspension were dropped in clean microscopic slides, air dried and stained with 10% Giemsa stain. One hundred well-spread metaphases were analyzed per mouse describing different kinds of chromosome abnormalities (CAs) in bone marrow and mouse spermatocytes. Scoring for CAs was performed under 2000× magnification with a light microscope. DNA fragmentation assay in mouse spleen cells 1. DNA gel electrophoresis laddering assay Apoptotic DNA fragmentation was qualitatively analyzed by detecting the laddering pattern of nuclear DNA as described according to Lu et al. [14]. Briefly, spleen tissues were homogenized, washed in PBS, and lysed in 0.5 ml of DNA extraction buffer (50 mM Tris–HCl, 10 mM EDTA. 0.5% Triton, and 100 μg/ml proteinase K, pH 8.0) for overnight at 37 °C. The lysate was then incubated with100 μg/ml DNase-free RNase for 2h at 37 °C, followed by three extractions of an equal volume of phenol/chloroform (1:1 v/v) and a subsequent re-extraction with chloroform by centrifuging at 15,000 rpm for 5 min at 4 °C. The extracted DNA was precipitated in two volume of ice-cold 100% ethanol with 1/10 volume of 3 M sodium acetate, pH 5.2 at −20 °C for 1h, followed by centrifuging at 15,000 rpm for 15 min at 4 °C. After washing with 70% ethanol, the DNA pellet was air-dried and dissolved in 10 mM Tris–HCl/1 mM EDTA, pH 8.0. The DNA was then electrophoresed on 1.5% agarose gel and stained with ethidium bromide in Tris/acetate/EDTA (TAE) buffer (pH 8.5, 2 mM EDTA, and 40 mM Tris–acetate). A 100-bp DNA ladder (Invitrogen, USA) was included as a molecular size marker and DNA fragments were visualized and photographed by exposing the gels to ultraviolet trans-illumination. 2. Diphenylamine reaction procedure Animal spleen tissues were used to determine the quantitative profile of the DNA fragmentation. Spleen samples were collected immediately after sacrificing the animals. The tissues were lysed in 0.5 ml of lysis buffer containing, 10 mM tris-HCl (pH 8), 1 mM EDTA, 0.2% triton X-100, centrifuged at 10 000 rpm (Eppendorf) for 20 min at 4°C. The pellets were re-suspended in 0.5 ml of lysis buffer. To the pellets (P) and the supernatants (S), 0.5 ml of 25% tri-chloroacetic acid (TCA) was added and incubated at 4°C for 24 h. The samples were then centrifuged for 20 min at 10 000 rpm (Eppendorf) at 4°C and the pellets were suspended in 80 ml of 5% TCA, followed by incubation at 83°C for 20 min. Subsequently, to each sample 160 ml of Diphenyl Amine (DPA) solution [150 mg DPA in 10 ml glacial acetic acid, 150 ml of sulfuric acid and 50 ml acetaldehyde (16 mg:ml)] was added and incubated at room temperature for 24h [15]. The proportion of fragmented DNA was calculated from absorbance reading at 600 nm wavelength using the formula: % Fragmented DNA = [OD(S)/[OD(S) + OD(P)] X 100 (OD: optical density, S: supernatants, P: pellets) Comet Assay in the testes Comet assay was performed referring to the protocol developed by Blasiak et al. [16] with minor modifications. Cells from testes of each treatment were mixed with low-melting-point agarose (ratio of1:10v/v), then pipetted to precoated slides with normal-melting-point agarose. The slides were kept flat at 4°C for 30 min in dark environment. The third layer of low melting point agarose was then pipetted on slides, left to solidify for 30 min at 4°C. The slides were transferred to pre-chilled lysis solution, kept for 60min at 4°C. After that, slides were immersed in freshly prepared alkaline unwinding solution at room temperature in the dark for 60 min. Slides were subjected to an electrophoresis run at 0.8 V/cm, 300mAmps at 4°C for 30 min. The slides were rinsed in neutralizing solution followed by immersion in 70% ethanol and then air-dried. Ethidium bromide was used for slides stain and then visualized by using Zeiss epifluorescence microscope (510–560 nm, barrier filter 590 nm) with a magnification of ×400. 100 cells per animal were scored then analyzed with DNA damage analysis software (Comet Score, TriTek corp., Sumerduck, VA22742). The nonoverlapping cells were randomly selected and were visually assigned a score on an arbitrary scale of 0–3 (i.e., class 0 = no detectable DNA damage and no tail; class 1 = tail with a length less than the diameter of the nucleus; class 2 = tail with length between 1× and 2× the nuclear diameter; and class 3 = tail longer than 2× the diameter of the nucleus) based on perceived comet tail length migration and relative proportion of DNA in the nucleus [17, 18]. Sperm shape abnormalities Sperm were prepared according to the recommended method of Wyrobek and Bruce [19] with some modifications recorded by Fahmy et al. [20] and smears were stained with 1% Eosin Y. A total of 1000 sperm were counted per animal (5000/each treatment), and different types of sperm abnormalities were scored (Head & Tail abnormalities). Sperm preparations were examined by light microscopy at 1000× magnification. Data analysis Data were analyzed using computerized software SPSS (Statistical Package of Social Science, version 20, Armonk, New York: IBM Corp). The data were checked for normality and the homogeneity of the variance using the Kolmogorov-Smirnov's test and Levene's test, respectively. The differences among groups with normal distribution were analyzed by one-way analysis of variance (ANOVA) followed by the Tukey HSD test. The results were regarded as significant when the P-value was less than or equal to 0.05. Results Chromosomal aberration analysis in bone marrow cells Table 1 presents the frequency of chromosomal aberrations (CAs) induced in mice bone marrow cells after treatment with VB. The results indicated a significant percentage of CAs with a dose-related relationship in both single and repeated treatments. With respect to the types of aberrations polyploidy metaphases represented the major percentage of aberrant cells followed by breakage and gap. The frequency of polyploidy reached 7.8% and 11.6% of the total counted metaphases after treatment with a single dose of 10 mg/kg and repeated doses of 6 mg/kg VB respectively. DNA damage in spleen tissues Rate of DNA fragmentation in spleen tissues of male mice exposed to different doses of vinblastine (3, 4.5, 6 and 10 mg) is presented in Table 2 and Figure 2. The results revealed that the dose 3 mg/kg VB had relatively similar rate of DNA fragmentation compared to control group. In contrast, the DNA fragmentation rate values were increased significantly and dose-dependent in spleen samples of mice exposed to other doses of VB compared to control group. Moreover, male mice exposed to 6 and 10 mg of VB exhibited higher DNA fragmentation compared to those exposed to 3 and 4.5 mg/kg VB. The highest DNA fragmentation was found in mice exposed to 10 mg/kg VB. Chromosomal aberration analysis in mouse spermatocytes Table 3 illustrates the percentage of different types of CAs induced in mouse spermatocytes after VB treatment. A significant and dose-dependent increase in the frequency of CAs was recorded. Repeated dose treatment gave a higher incidence of CAs in comparing with the single dose. The majority of aberrations were in the form of univalent (X-Y and Autosomal univalent) with the superiority of X-Y univalent in comparison with autosomal univalent. The maximum percentage of CAs (13.2 %) was reached after single dose treatment at 10 mg /kg VB. Alkaline comet assay in testes tissues The DNA damage in the testes tissues of male mice exposed to a single dose of vinblastine (VB) at 3, 4.5, 6 and 10 mg/kg is summarized in Table 4 and Figure 3. Group of mice exposed to the low dose of VB (3 mg) showed relatively similar rate of DNA damage compared to control group. The values of DNA damage were increased significantly in other doses of VB compared to control group. Dose-related relationship was recorded. Additionally, the highest DNA damage of class 3 (tail longer than 2× the diameter of the nucleus) was showed in mice exposed to 10 mg/kg VB. Morphological sperm abnormalities The results in Table 5 demonstrated a significant (p˂ 0.05) and dose-dependent increase in the frequency of sperm abnormalities in comparison with the negative control. The maximum percentage of sperm defects reached 11.06% after treatment with the dose of 6 mg/kg vs 3.04% for the control. With respect to the types of sperm abnormalities, Coiled tail sperm represents the major percentage. Shapeless head sperm and head without hook were also recorded. In the highest tested dose, sperm tail defects represent 7.66 % while the head abnormalities represent 3.4% of the total counted sperm. Discussion Chemotherapy represents the main therapeutic choice in treating cancer. The majority of chemotherapies have no selectivity to cancer cells and affect healthy body tissues where the cells are constantly growing and dividing. This fact can be explained by the side effects associated with the course of chemotherapeutic regimens. Useful drugs without side effects do not yet exist, so it is of a great importance to estimate the side effects of chemotherapy and the data must be available for patients, doctors and the pharmaceutical companies to reach the safe use of chemotherapy. However, in cancer survivors, a second malignancy frequently occurs after chemotherapy, which warrants detailed genotoxicity testing of the chemotherapeutic agents [ 5 ]. Herein, a comprehensive study was performed to evaluate the genotoxic effects of vinblastine (VB) using several endpoints as biomarkers: Chromosomal aberrations in bone marrow and mice spermatocytes, DNA fragmentation in somatic and germ cells and sperm abnormalities. VB is one of the Vinca alkaloid family which represents antitumor agents from plants. From Vinca alkaloids, VB and vincristine (VC) have been well established in the treatment of cancer. The results showed that VB induced a significant percentage of chromosomal aberrations in somatic cells (bone marrow) which was dose-dependent in both single and repeated treatments. The frequency of chromosomal aberrations reached about 4-folds increase after treatment with a single dose of 10 mg/kg and about 6-folds increase after 3 successive doses at 6 mg/kg of VB as compared with the negative control. A pronounced number of polyploidy was recorded at all doses and also a number of metaphases with fragments (numerical and structural aberrations). Vinca alkaloids appear to exert their antitumor effect by binding to tubulin proteins within the cells. Because these proteins are essential contractile proteins of the mitotic spindle of dividing cells, this binding leads to mitotic arrest [ 7 ] which may explain the appearance of a high percentage of polyploidy after VB treatment. Generally, in the absence of an intact mitotic spindle, the chromosomes may clump in an unusual grouping or may disperse throughout the cytoplasm [ 21 ]. Previous studies with somatic cell cultures revealed the presence of aneuploidy and polyploidy after VB treatment [ 22 , 23 ]. Due to their action as mitotic spindle poisons, vinca alkaloids are regarded as aneugens [ 24 ]. Vinca alkaloids are also cell cycle phase-specific ´S and M´ phase poisons. The present study also demonstrated DNA damage in spleen cells which also recorded a dose-dependent relationship. The effect of VB in somatic cells is coincided well with the previous results of other authors: micronuclei and chromosomal aberrations were significantly induced in mouse bone marrow after treatment with VB [ 4 , 25 , 26 ] and in cultured human lymphocytes after VC treatment [ 27 ]. However, the induction of micronuclei may reflect chromosomal damage or loss of whole chromosomes from the daughter nuclei. Oxidative DNA damage in cultured human lymphocytes and an increase in the cells mitotic index ( p < 0.05) were also demonstrated after VB and VC treatment [ 2 ]. They also induced chromosomal mutations in in vivo and in cultured cancer cells [ 28 , 29 ]. Moreover, VB is found to affect nucleic acids, amino acids, proteins and purine synthesis [ 30 ]. Tyagi et al. [ 31 ] made a spectroscopic study showed that plant alkaloid binds with DNA via A-T and G-C base pairs along with the phosphate back bone of the helix. In addition, the docking studies have indicated the interaction of VB with the adenine base of DNA helix via hydrogen bonding. Conversely, other studies showed lack of mutagenic activity of Vinca alkaloids [ 32 , 33 ]. Generally, aneugenic agents are expected to have a low level of clastogencity as a result of their poisoning effect on the mitotic spindle [ 34 ]. Chromosomal aberration analysis was also examined in mouse spermatocytes. The current results indicated a significant percentage of chromosomal aberrations (p ˂ 0.05) after single and repeated treatments with the presence of dose-relationship. The maximum percentage of aberrations reached approximately 3.5 and 3.2-folds increase as compared to the negative control after single dose of 10 mg/kg and repeated doses of 6 mg/kg VB respectively. The majority of aberrations were X-Y and Autosomal univalent (gonosomal univalency) and fragments. Induction of significant percentages of aberrant primary spermatocytes (P ≤ 0.01) was also recorded by Palo et al. [ 8 ] with atypical bivalents. Moreover, the current results indicated DNA damage in the testes as measured by alkaline comet assay and evidenced by comet tail length. Such effect may be as a result of oxidative stress exerted by VB in the form of reactive oxygen and nitrogen species (ROS/RNS) and to its depleted activity on the antioxidant enzymes: SOD, CAT and GPx. VB was also reported to induce a disturbance in the intracellular mediator’s ´H 2 O 2 and Ca 2+ level´ [ 2 , 35 ]. In addition, it was demonstrated to alter calcium homeostasis via mitochondrial membranes leading to cytotoxicity and chromosome instability. Inducing of apoptosis is considered one of its mechanisms of action. The present results are also concerned with measuring the morphological sperm abnormalities after VB treatment (at different doses). Different categories of abnormal sperm were noticed and indicated the transmission of VB-induced cytogenotoxic effects from spermatocyte to sperm. Jagetia et al. [ 36 ] demonstrated that different doses of VB can affect mouse spermatogenesis as measured by DNA flow cytometry. Previous reports also showed significant percentages of aberrant spermatogonial metaphases and chromosomal aberrations in VB-treated mice. VB also significantly increased the percentages of aberrant primary spermatocytes and the morphological sperm defects as indicated by Palo et al. [ 8 ]. This result in addition to the results of the current work emphasizes the ability of VB to cause genetic defects that may be passed on to future generations. The cancer survivors who are treated with chemotherapy before or during their reproductive years have faced serious problem. Such effect must be taken into consideration when choosing the chemotherapeutic regimens which may jeopardize the genetic health of offspring [ 37 ]. Infertility is another risk problem that faces cancer survivors. Therapeutic modalities such as radiation therapy and chemotherapy are highly effective in treating cancer, but their gonadotoxic side effects can severely impair fertility in an agent- and dose-dependent way. Spermatogenesis for example in long-term cancer survivors has elucidated evidence of persistent azoospermia or severe oligozoospermia in up to 24% of patients [ 38 ]. This effect warrants several years after chemotherapy to recover. The alkylating agents such as cyclophosphamide and isophosphamide caused permanent azoospermia in 80–90% of cases [ 39 ]. Azoospermia was detected follow the use of VB in testicular cancer patients and it may be reversible within 2–3 years [ 40 ]. In the present study VB induced a high percentage of sperm defects (tail and head defects) with a pronounced appearance of coiled tail sperm. Also the absence of acrosome and amorphous or misshape head sperm was recorded. These defects might affect the ability of the sperm to reach and penetrate an egg. Coiling of the sperm tail may limit or stop its motility and also acrosome plays a crucial role in its function. Sperm motility and acrosome reaction are considered key functions in the control of reproduction and also they are essential for spermatozoa to become fertile [ 41 ]. The sperm morphology was reported to be genetically controlled by numerous autosomal and sex-linked genes [ 42 ] and the formation of a normal sperm head involves intricate synchronous morphological and biochemical steps [ 43 ]. Oxidative stress, high levels of lipid peroxidation and oxidative DNA damage accompanied by mutation were considered the major causes of male infertility. Mutations that influence sperm quality include conditions that affect the morphological appearance of spermatozoa and their competence of fertilization [ 44 ]. The current results emphasize the positive correlation between cytogenetic damage in germ cells and sperm abnormalities. Conclusion The present work demonstrated that vinblastine has a genotoxic effect evidenced by DNA damage and chromosomal aberrations (CAs) induced in both somatic and germ cells of male mice. In bone marrow cells CA analysis revealed a pronounced number of polyploidy metaphases which reflect the effect of VB in the mitotic spindle. CAs induced by VB in spermatocytes were transmitted to sperm causing several defect categories. Such defects are expected to affect the fertility of the cancer survivors or might be result in malformation in the future offspring. The results of the present work must be taken into consideration while using VB in the chemotherapeutic regimens. 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Mutation Research/Genetic Toxicology and Environmental Mutagenesis 471: 29-36. https://doi.org/10.1016/s1383-5718(00)00115-7 Morales-Ramírez P, Vallarino-Kelly T, Cruz-Vallejo V (2004) Kinetics of micronucleated polychromatic erythrocyte (MN-PCE) induction in vivo by aneuploidogens, Mutation Research/Genetic Toxicology and Environmental Mutagenesis 565: 79-87. https://doi.org/10.1016/j.mrgentox.2004.09.006 Jones R, Richards J, Beer C (1966) Biochemical studies with the vinca alkaloids: ii. Effect of vinblastine on the biosynthesis of nucleic acids and their precursors in rat thymus cells. Cancer Research 26: 882-887. Tyagi G, Charak S, Mehrotra R (2012) Binding of an indole alkaloid, vinblastine to double stranded DNA: A spectroscopic insight in to nature and strength of interaction. Journal of Photochemistry and Photobiology B: Biology 108: 48-52. https://doi.org/10.1016/j.jphotobiol.2011.12.009 Gundy S, Baki M, Bodrogi I (1989) Vinblastine, cisplatin and bleomycin (VPB) adjuvant therapy does not induce dose-dependent damage in human chromosomes. Neoplasma 36: 457-464. González-Cid M, Cuello MT, Larripa I (1997) Mitotic arrest and anaphase aberrations induced by vinorelbine in hamster cells in vitro. Anti-cancer drugs 8: 529-532. https://doi.org/10.1097/00001813-199706000-00017 Warr TJ, Parry EM, Parry J (1993) A comparison of two in vitro mammalian cell cytogenetic assays for the detection of mitotic aneuploidy using 10 known or suspected aneugens. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 287: 29-46. https://doi.org/10.1016/0027-5107(93)90143-4 Rtibi K, Grami D, Selmi S, Amri M, Sebai H, Marzouki L (2017) Vinblastine, an anticancer drug, causes constipation and oxidative stress as well as others disruptions in intestinal tract in rat. Toxicology Reports 4: 221-225. https://doi.org/10.1016/j.toxrep.2017.04.006 Jagetia GC, Krishnamurthy H, Jyothi P (1996) Evaluation of cytotoxic effects of different doses of vinblastine on mouse spermatogenesis by flow cytometry. Toxicology 112: 227-236. https://doi.org/10.1016/0300-483X(96)03402-6 Wyrobek AJ, Schmid TE, Marchetti F (2005) Relative susceptibilities of male germ cells to genetic defects induced by cancer chemotherapies. JNCI Monographs 31-35. https://doi.org/10.1093/jncimonographs/lgi001 lopez Andreu JA, Fernandez PJ, Tortajada JF, Navarro I, Rodriguez-Ineba A, Muro MD, Romeu A (2000) Persistent altered spermatogenesis in long-term childhood cancer survivors. Pediatric Hematology and Oncology 17: 21-30. https://doi.org/10.1080/088800100276631 Pryzant RM, Meistrich ML, Wilson G, Brown B, McLaughlin P (1993) Long-term reduction in sperm count after chemotherapy with and without radiation therapy for non-Hodgkin's lymphomas. Journal of Clinical Oncology 11: 239-247. https://doi.org/10.1200/JCO.1993.11.2.239 Fosså SD, Theodorsen L, Norman N, Aabyholm T (1990) Recovery of impaired pretreatment spermatogenesis in testicular cancer. Fertility and Sterility 54: 493-496. https://doi.org/10.1016/s0015-0282(16)53768-6 Visconti P, Westbrook V, Chertihin O, Demarco I, Sleight S, Diekman A (2002) Novel signaling pathways involved in sperm acquisition of fertilizing capacity. Journal of Reproductive Immunology 53: 133-150. https://doi.org/10.1016/s0165-0378(01)00103-6 Krzanowska H (1976) Inheritance of sperm head abnormality types in mice–the role of the Y chromosome. Genetics Research 28: 189-198. https://doi.org/10.1017/S0016672300016864 Rattner J (1972) Nuclear shaping in marsupial spermatids. J Ultrastruct Res 40: 498-512. https://doi.org/10.1016/s0022-5320(72)80038-8 Aitken RJ, Baker MA (2020) The role of genetics and oxidative stress in the etiology of male infertility-A unifying hypothesis? Front. Endocrinol 11: 581838. https://doi.org/10.3389/fendo.2020.581838 Tables Table1. Frequency of chromosomal aberrations induced in mouse bone marrow cells after treatment with vinblastine. Treatment and doses Total abnormal metaphases No and (%) of metaphases with different types of chromosome aberrations No. Mean(%) ± SE Gap Fragment and/or Break Polyploidy I. Control (Non- treated) 16 3.20 ± 0.37 a 7(1.40) 9(1.80) - II. Vinblastine a-single dose 3 mg/kg 32 6.40 ± 0.60 b 6(1.20) 10(2.0) 16(3.20) 4.5 mg/kg 41 8.20 ± 0.58 b,c 5(1.0) 12(2.40) 24(4.80) 6 mg/kg 51 10.20 ± 0.37 c,d 6(1.20) 20(4.0) 25(5.0) 10 mg /kg 60 12.0 ± 0.95 d,e 8(1.60) 13(2.60) 39(7.80) b- 3 successive dose treatment 3 mg/kg 65 13.0± 0.95 e 8(1.60) 12(2.40) 45(9.0) - 4.5 mg/kg 83 16.60 ± 0.93 f 11(2.20) 22(4.40) 50(10.0) - 6 mg/kg 97 19.40 ± 0.56 g 14(2.80) 25(5.0) 58(11.60) A total of 500 cells were analyzed (5 mice per group; 100 cells/mouse). One way ANOVA–Tukey’s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another. Table2. DNA fragmentation detected in spleen tissues of mice exposed to different doses vinblastine. Treatment and doses DNA Fragmentation % (M ± SEM) Change Inhibition % (-ve) control 9.5±0.71 a 0.00 0.00 Vinblastine 3 mg 10.2±0.49 ab 0.70 95.24 4.5 mg 12.4±0.47 c 2.90 80.27 6 mg 18.6±0.67 dc 9.10 38.09 10 mg 24.2±0.88 d 14.7 29.28 Means with different superscripts ( a, b,c,d ) between groups in the same column are significantly different at P<0.05. Table3. Frequency of chromosomal aberrations induced in mouse spermatocytes after treatment with vinblastine. Treatment and doses Total abnormal metaphases No and (%) of metaphases with different types of chromosome aberration No. Mean(%) ± SE X-Y univalent Autosomal univalent X-y u. + A-u. Fragment and/or Break Chain IV Polyp. I. Control (Non-treated) 19 3.80 ± 0.37 a 17(3.40) 2(0.40) - - - - II. Vinblastine a- Single dose 3 mg/kg 40 8.0 ± 0.77 b 25(5.0) 12(2.40) - 1(0.20) - 2(0.40) 4.5 mg/kg 44 8.80 ± 0.37 b,c 28(5.60) 12(2.40) - 1(0.20) - 3(0.60) 6 mg/kg 52 10.40 ± 0.93 c,d 23(4.60) 20(4.0) 1(0.20) 3(0.60) 1(0.20) 4(0.80) 10 mg /kg 66 13.20 ± 1.16 e 45(9.0) 17(3.40) 2(0.40) 2(0.20) - - b - 3 successive dose treatment 3 mg/kg 51 10.20 ± 0.86 b,c,d 27(5.40) 18(3.60) - 4(0.40) 1(0.20) 1(1.0) 4.5 mg/kg 55 11.0 ± 0.83 c,d,e 41(8.20) 8(1.60) - 5(1.0) - 1(1.0) 6 mg/kg 61 12.20 ± 0.37 d,e 30(6.0) 18(3.60) - 6(1.20) - 7(1.40) A total of 500 cells were analyzed (5 mice per group; 100 cells/mouse). X-Y u: X-Y univalent, A-u: Autosomal univalent, Polyp: Polyploidy. One way ANOVA–Tukey’s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another. Table 4. Visual score of DNA damage in the testes of male mice exposed to different doses of vinblastine. Treatment No of samples No. of cells Class ** DNA damaged cells % (Mean ± SE) Analyzed * Comets 0 1 2 3 I- (-ve) control 5 500 36 464 25 11 0 7.21±1.07 a II- Vinblastine 3 mg/kg 5 500 39 461 23 16 0 7.83±0.93 b 4.5 mg/kg 5 500 51 449 28 17 6 10.24±1.24 c 6 mg/kg 5 500 83 417 31 28 24 16.61±1.50 cd 10 mg/kg 5 500 107 393 35 31 41 21.43±1.21 d *: Number of cells examined per a group, ** : Class 0= no tail; 1= tail length < diameter of nucleus; 2= tail length between 1×, and 2× the diameter of nucleus; and 3= tail length > 2× the diameter of nucleus. Data are presented as Mean ± SE. Mean values within tissue with unlike superscript letters were significantly different ( P <0.05). Table5. Percentage of sperm abnormalities induced in male mice after treatment with vinblastine Experimental groups and doses Total abnormal sperm No. and (%)of different types of sperm abnormalities Head abnormalities Tail abnormalities No. Mean (%)±SE Amorphous Without hook Triangle Banana Coiled tail I- Control (-ve) 152 3.04 ± 0.19 a 16(0.32) 4(0.08) 2(0.04) 1(0.02) 20(0.40) II- Vinblastine 3 mg/kg 472 9.44 ±0.13 b 71(1.42) 69(1.38) 18(0.36) 23(0.46) 291(5.82) 4.5 mg/kg 520 10.40 ± 0.73 b,c 85(1.70) 67(1.34) 19(0.38) 19(0.38) 330(6.60) 6mg/kg 553 11.06±0.14 c 54(1.08) 87(1.74) 18(0.36) 11(0.22) 383(7.66) Total number of examined sperms 5000 per each treatment (1000 /mouse, 5 mice/group) .One way ANOVA–Tukey’s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another at p<0.05. Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 12 Aug, 2022 Reviewers invited by journal 12 Aug, 2022 Editor assigned by journal 09 Aug, 2022 First submitted to journal 08 Aug, 2022 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-1943872\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":128477877,\"identity\":\"7a82d3f8-ed6d-4981-b4d5-46f841e07e83\",\"order_by\":0,\"name\":\"Maha A. Fahmy\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"National Research Centre\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Maha\",\"middleName\":\"A.\",\"lastName\":\"Fahmy\",\"suffix\":\"\"},{\"id\":128477878,\"identity\":\"7c9b3e67-ad10-4bfd-a05c-3abdd47b444d\",\"order_by\":1,\"name\":\"Entesar Hassan\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8UlEQVRIiWNgGAWjYBCDBAhVwcBgQFgxM7KWMyRrYWwjQgu/dP/Bhz9q7uXx9x9/+Lhw3mF5c/bmAww/Krbh1CI55zCzMc+x4mKJGznGxjO3HTbc2XMsgbHnzG2cWgxuJLNJM7AlJDbc4GGT5t12mHHDjRwDZsY23FrsbySz//zxLyFx/vnjz3/zzjlsT1CLgUQyGwNvW0LihgMJZsy8DYcTCWqRuJFsLM3bl5C4EegXaZ5j6ckbzhxLOIjPL/wzEh9+/PEtIXHe+eMPP/PUWNtuON588MGPCtxa0EEzmDxAtHogqCNF8SgYBaNgFIwQAACJv1y+eG58DAAAAABJRU5ErkJggg==\",\"orcid\":\"https://orcid.org/0000-0001-7999-4797\",\"institution\":\"National Research Centre\",\"correspondingAuthor\":true,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Entesar\",\"middleName\":\"\",\"lastName\":\"Hassan\",\"suffix\":\"\"},{\"id\":128477879,\"identity\":\"7b648922-a736-44ae-a168-93739dbd414b\",\"order_by\":2,\"name\":\"Ayman A. Farghaly\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"National Research Centre\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Ayman\",\"middleName\":\"A.\",\"lastName\":\"Farghaly\",\"suffix\":\"\"},{\"id\":128477880,\"identity\":\"771357db-e07c-462b-842a-dcb66598b170\",\"order_by\":3,\"name\":\"Zeinab M. Hassan\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"National Research Centre\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Zeinab\",\"middleName\":\"M.\",\"lastName\":\"Hassan\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2022-08-09 07:03:03\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-1943872/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-1943872/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":25339510,\"identity\":\"3122752c-3e74-453b-bb9b-018dd05cc751\",\"added_by\":\"auto\",\"created_at\":\"2022-08-17 20:36:26\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":10448,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eRepresents the chemical structure of vinblastine: C\\u003csub\\u003e46\\u003c/sub\\u003eH\\u003csub\\u003e58\\u003c/sub\\u003eN\\u003csub\\u003e4\\u003c/sub\\u003eO\\u003csub\\u003e9 \\u003c/sub\\u003e(PubChem).\\u003c/p\\u003e\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-1943872/v1/95ab56253b67c3f7092f2129.png\"},{\"id\":25339511,\"identity\":\"fa23d4f8-f813-4682-b752-3a96d62fee03\",\"added_by\":\"auto\",\"created_at\":\"2022-08-17 20:36:26\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":79933,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDNA fragmentation detected with agarose gel of DNA extracted from spleen tissues of mice exposed to different doses of vinblastine.\\u0026nbsp;Lane M represents DNA marker. Lane 1 represents negative control group. Lane 2, 3, 4 and 5 represent mice treated with the doses 3, 4.5, 6, and 10 mg/kg vinblastine respectively.\\u003c/p\\u003e\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-1943872/v1/24ff5fce92428e508ac07aa2.png\"},{\"id\":25339509,\"identity\":\"29b28a78-9cc6-45e2-ada7-93225a41cd0b\",\"added_by\":\"auto\",\"created_at\":\"2022-08-17 20:36:26\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":93095,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eVisual score of normal DNA (class 0) (a), cell with DNA damage (classes 1 and 2) (b), cell with DNA damage (class 3) (c), using comet assay in mice testes tissue collected from\\u0026nbsp;vinblastine treated groups.\\u003c/p\\u003e\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-1943872/v1/d730312082e490a61d8e73de.png\"},{\"id\":25339512,\"identity\":\"51005abb-f375-4b80-8fda-66e9b52323eb\",\"added_by\":\"auto\",\"created_at\":\"2022-08-17 20:36:29\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":541293,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-1943872/v1/77c1ad7c-edcc-42b6-adf3-7fcf64790285.pdf\"}],\"financialInterests\":\"\",\"formattedTitle\":\"Genotoxicity, DNA damage and sperm defects induced by vinblastine\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eVinblastine (VB) is a very important clinic alkaloid extracted naturally from a periwinkle plant which is a flowering herb from \\u003cem\\u003eCatharanthus roseus\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. It is a chemotherapeutic medication used separately or in combination with other cytotoxic drugs (bleomycin, doxorubicin, and methotrexate\\u0026hellip;) for the treatment of a wide range of human neoplasms \\u003cem\\u003ee.g.\\u003c/em\\u003e Hodgkin and non-Hodgkin\\u0026rsquo;s lymphoma, T-cell lymphoma, testicular and germ cell tumors, bladder, lung (Non-small cell lung cancer), head and neck cancers. It is also used to treat melanoma, soft tissue sarcoma, Kaposi\\u0026rsquo;s sarcoma, gestational trophoblastic and some blood disorders (\\u003cem\\u003ee.g.\\u003c/em\\u003e histiocytosis X) [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e]. In some cases it exerts an immunosuppressive effects. VB is one of the World Health Organization List of Essential Medications. It is a member of the Vinca alkaloid family which are anticancer agents that include the natural product vinblastine, vincristine, and the semi-synthetic product vin-orelbine [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. Vinca alkaloids have the potential to induce cytotoxic and genotoxic effects, oxidative DNA damage and micronuclei formation [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. Vinca alkaloids exert their function as mitotic spindle inhibitors [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e].They are antimitotic drugs that inhibit microtubules and are cell cycle phase-specific \\u0026acute;S and M phase poisons\\u0026acute; [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e]. VB is potentially mutagenic and animal studies have demonstrated degenerative changes in germ cells [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e], and reported an inhibition of angiogenic action [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eA major problem with cancer chemotherapy is its severe toxic effect on non-target tissues (normal cells), so the cancer patients not only suffer from an immediate side effects of cancer medications but are also subjected to an increased risk of developing secondary tumors later in life [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e]. Recent studies in chemotherapy approaches claim at developing medications that eradicate cancer cells efficiently and selectively. So it is of special need to study the side effects of anticancer agents in order to improve the conventional chemotherapy by protecting healthy tissues. The genotoxic reports concerning VB are controversial, limited and inconclusive. The aim of the present work is to determine the genotoxic effects of VB using different mutagenic biomarkers: chromosomal aberration analysis and DNA damage in both somatic and germ cells, and also assessment of sperm defects induced in male mice.\\u003c/p\\u003e\"},{\"header\":\"Materials And Methods\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eChemicals\\u0026nbsp;\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eCommon trade name of the chemotherapeutic drug is vinblastine sulfate, velban and others.\\u0026nbsp;Vinblastine\\u0026nbsp;was obtained from Sigma Aldrich (MO,USA).\\u0026nbsp;Drug Bank Accession Number: DB00570. All other chemicals were purchased from ADWICC (Cairo, Egypt).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAnimals\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMale white Swiss mice (\\u003cem\\u003eMus musculus\\u003c/em\\u003e), aged 9 to 12 weeks, were used in all experiments. Animals were obtained from a closed random-bred colony at the National Research Centre (Giza, Egypt). Mice used for each experiment were of similar age (\\u0026plusmn;1 week) and weight (\\u0026plusmn; 2 g). Animals were housed in polycarbonate boxes with steel wire tops and bedded with wood shavings. Ambient temperature was controlled at 22\\u0026deg;C \\u0026plusmn; 3\\u0026deg;C with a relative humidity of 50% \\u0026plusmn; 15% and a 12 h light/dark photoperiod. Food and water were provided \\u003cem\\u003ead libitum\\u003c/em\\u003e. The experiments were conducted according to the Animal Research Ethical Committee Guidelines of the National Research Centre, Egypt. The Approval Certificate is under number: 19 163.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eExperimental design\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eIn these experiments a total of 85 mice were taken as follows: 40 mice for chromosomal aberrations in bone marrow and mouse spermatocytes, 25 mice for DNA fragmentation and alkaline comet assay and 20 for morphological sperm abnormalities. In each of these testes, mice were subdivided into groups (5 /group). \\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThe main groups represented chromosomal aberration analysis: Group I: Negative control, Groups II-V, mice were i.p injected with a single dose of VB (3, 4.5, 6 and 10 mg/kg), Groups VI-VIII in which mice received repeated i.p injections (three successive days) of VB at the dose levels 3, 4.5 and 6 mg/kg. In all experiments samples were taken 24h after the last treatment.\\u003c/p\\u003e\\n\\u003cp\\u003eAnalysis for DNA damage, showed five groups: Animals were treated with a single i.p injection of VB at the dose levels 3, 4. 5, 6 and 10 mg/kg, in addition to the control group. Samples were taken 24h after injection.\\u003c/p\\u003e\\n\\u003cp\\u003eAnalysis of sperm abnormalities showed four groups as follows: Negative control group and three treated groups with VB (3, 4.5 and 6 mg/kg, 3 injections) and samples were taken 35 days after the 1\\u003csup\\u003est\\u003c/sup\\u003e injection. Different doses were taken to cover all doses required for different types of human cancerous.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCytogenetic analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eChromosomal aberration assay in mouse bone marrow and spermatocytes\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMitotic and meiotic chromosomes were prepared from bone marrow and testis of the same animal, respectively. Bone marrow chromosomes were prepared according to the technique described by Diab et al. [11]. In brief, mouse bone- marrow cells were collected from both femurs, cells were incubated in hypotonic solution (KCL 0.075 M) for 20 min at 37\\u0026deg;C, and then centrifuged. The cell pellets were suspended in a fixative (methanol/glacial acetic acid 3:1). This step was repeated at least twice, then the cells were suspended in a few drops of fixative and spread onto frozen slides, air-dried, stained with 10% Giemsa for 30 min, washed, and air dried again.\\u003c/p\\u003e\\n\\u003cp\\u003eSpermatocyte chromosomes were prepared from the testes according to the protocol described by Evans et al.[12]with some modifications [13]. Briefly, the testis was removed and squashed into a petri dish containing an isotonic solution of 2.2% trisodium citrate. Then the cell suspension was centrifuged for 5 minutes at 1500 rpm. The cell pellet was incubated in a hypotonic solution of 1.1% trisodium citrate for 20 minutes at 37C◦ followed by centrifugation. The cell pellet was washed twice by a freshly prepared fixative. A few drops of the fixative cell suspension were dropped in clean microscopic slides, air dried and stained with 10% Giemsa stain. \\u0026nbsp;One hundred well-spread metaphases were analyzed per mouse describing different kinds of chromosome abnormalities (CAs) in bone marrow and mouse spermatocytes. Scoring for CAs was performed under 2000\\u0026times; magnification with a light microscope.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eDNA fragmentation assay in mouse spleen cells\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e1. DNA gel electrophoresis laddering assay\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eApoptotic DNA fragmentation was qualitatively analyzed by detecting the laddering pattern of nuclear DNA as described according to Lu et al. [14]. Briefly, spleen tissues were homogenized, washed in PBS, and lysed in 0.5 ml of DNA extraction buffer (50 mM Tris\\u0026ndash;HCl, 10 mM EDTA. 0.5% Triton, and 100 \\u0026mu;g/ml proteinase K, pH 8.0) for overnight at 37 \\u0026deg;C. The lysate was then incubated with100 \\u0026mu;g/ml DNase-free RNase for 2h at 37 \\u0026deg;C, followed by three extractions of an equal volume of phenol/chloroform (1:1 v/v) and a subsequent re-extraction with chloroform by centrifuging at 15,000 rpm for 5 min at 4 \\u0026deg;C. The extracted DNA was precipitated in two volume of ice-cold 100% ethanol with 1/10 volume of 3 M sodium acetate, pH 5.2 at \\u0026minus;20 \\u0026deg;C for 1h, followed by centrifuging at 15,000 rpm for 15 min at 4 \\u0026deg;C. After washing with 70% ethanol, the DNA pellet was air-dried and dissolved in 10 mM Tris\\u0026ndash;HCl/1 mM EDTA, pH 8.0. The DNA was then electrophoresed on 1.5% agarose gel and stained with ethidium bromide in Tris/acetate/EDTA (TAE) buffer (pH 8.5, 2 mM EDTA, and 40 mM Tris\\u0026ndash;acetate). A 100-bp DNA ladder (Invitrogen, USA) was included as a molecular size marker and DNA fragments were visualized and photographed by exposing the gels to ultraviolet trans-illumination.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2. Diphenylamine reaction procedure\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAnimal spleen tissues were used to determine the quantitative profile of the DNA fragmentation. Spleen samples were collected immediately after sacrificing the animals. The tissues were lysed in 0.5 ml of lysis buffer containing, 10 mM tris-HCl (pH 8), 1 mM EDTA, 0.2% triton X-100, centrifuged at 10 000 rpm (Eppendorf) for 20 min at 4\\u0026deg;C. The pellets were re-suspended in 0.5 ml of lysis buffer. To the pellets (P) and the supernatants (S), 0.5 ml of 25% tri-chloroacetic acid (TCA) was added and incubated at 4\\u0026deg;C for 24 h. The samples were then centrifuged for 20 min at 10 000 rpm (Eppendorf) at 4\\u0026deg;C and the pellets were suspended in 80 ml of 5% TCA, followed by incubation at 83\\u0026deg;C for 20 min. Subsequently, to each sample 160 ml of Diphenyl Amine (DPA) solution [150 mg DPA in 10 ml glacial acetic acid, 150 ml of sulfuric acid and 50 ml acetaldehyde (16 mg:ml)] was added and incubated at room temperature for 24h [15]. The proportion of fragmented DNA was calculated from absorbance reading at 600 nm wavelength using the formula:\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e% Fragmented DNA = [OD(S)/[OD(S) + OD(P)] X 100\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;(OD: optical density, S: supernatants, P: pellets)\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eComet Assay in the testes\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eComet assay was performed referring to the protocol developed by Blasiak et al. [16] with minor modifications. Cells from testes of each treatment were mixed with low-melting-point agarose (ratio of1:10v/v), then pipetted to precoated slides with normal-melting-point agarose. The slides were kept flat at 4\\u0026deg;C for 30 min in dark environment. The third layer of low melting point agarose was then pipetted on slides, left to solidify for 30 min at 4\\u0026deg;C. The slides were transferred to pre-chilled lysis solution, kept for 60min at 4\\u0026deg;C. After that, slides were immersed in freshly prepared alkaline unwinding solution at room temperature in the dark for 60 min. Slides were subjected to an electrophoresis run at 0.8 V/cm, 300mAmps at 4\\u0026deg;C for 30 min. The slides were rinsed in neutralizing solution followed by immersion in 70% ethanol and then air-dried. Ethidium bromide was used for slides stain and then visualized by using Zeiss epifluorescence microscope (510\\u0026ndash;560 nm, barrier filter 590 nm) with a magnification of \\u0026times;400. 100 cells per animal were scored then analyzed with DNA damage analysis software (Comet Score, TriTek corp., Sumerduck, VA22742). The nonoverlapping cells were randomly selected and were visually assigned a score on an arbitrary scale of 0\\u0026ndash;3 (i.e., class 0 = no detectable DNA damage and no tail; class 1 = tail with a length less than the diameter of the nucleus; class 2 = tail with length between 1\\u0026times; and 2\\u0026times; the nuclear diameter; and class 3 = tail longer than 2\\u0026times; the diameter of the nucleus) based on perceived comet tail length migration and relative proportion of DNA in the nucleus [17, 18].\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;\\u003cstrong\\u003eSperm shape abnormalities\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eSperm were prepared according to the recommended method of Wyrobek and Bruce [19] with some modifications recorded by Fahmy et al. [20] and smears were stained with 1% Eosin Y. A total of 1000 sperm were counted per animal (5000/each treatment), and different types of sperm abnormalities were scored (Head \\u0026amp; Tail abnormalities). Sperm preparations were examined by light microscopy at 1000\\u0026times; magnification.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eData were analyzed using computerized software SPSS (Statistical Package of Social Science, version 20, Armonk, New York: IBM Corp). The data were checked for normality and the homogeneity of the variance using the Kolmogorov-Smirnov\\u0026apos;s test and Levene\\u0026apos;s test, respectively. The differences among groups with normal distribution were analyzed by one-way analysis of variance (ANOVA) followed by the Tukey HSD test. The results were regarded as significant when the P-value was less than or equal to 0.05.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eChromosomal aberration analysis in bone marrow cells\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eTable 1 presents the frequency of chromosomal aberrations (CAs) induced in mice bone marrow cells after treatment with VB. The results indicated a significant percentage of CAs with a dose-related relationship in both single and repeated treatments. With respect to the types of aberrations polyploidy metaphases represented the major percentage of aberrant cells followed by breakage and gap. The frequency of polyploidy reached 7.8% and 11.6% of the total counted metaphases after treatment with a single dose of 10 mg/kg and repeated doses of 6 mg/kg VB respectively.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eDNA damage in spleen tissues \\u0026nbsp;\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eRate of DNA fragmentation in spleen tissues of male mice exposed to different doses of vinblastine (3, 4.5, 6 and 10 mg) is presented in Table 2 and Figure 2. The results revealed that the dose 3 mg/kg VB had relatively similar rate of DNA fragmentation compared to control group. In contrast, the DNA fragmentation rate values were increased significantly and dose-dependent in spleen samples of mice exposed to other doses of VB compared to control group. Moreover, male mice exposed to 6 and 10 mg of VB exhibited higher DNA fragmentation compared to those exposed to 3 and 4.5 mg/kg VB. The highest DNA fragmentation was found in mice exposed to 10 mg/kg VB.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eChromosomal aberration analysis in mouse spermatocytes\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eTable 3 illustrates the percentage of different types of CAs induced in mouse spermatocytes after VB treatment. A significant and dose-dependent increase in the frequency of CAs was recorded. Repeated dose treatment gave a higher incidence of CAs in comparing with the single dose. The majority of aberrations were in the form of univalent (X-Y and Autosomal univalent) with the superiority of X-Y univalent in comparison with autosomal univalent. The maximum percentage of CAs (13.2 %) was reached after single dose treatment at 10 mg /kg VB.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAlkaline comet assay in testes tissues \\u0026nbsp; \\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe DNA damage in the testes tissues of male mice exposed to a single dose of vinblastine (VB) at 3, 4.5, 6 and 10 mg/kg is summarized in Table 4 and Figure 3.\\u0026nbsp;Group of mice exposed to the low dose of\\u0026nbsp;VB (3 mg) showed relatively similar rate of DNA damage compared to control group. The values of\\u0026nbsp;DNA damage were increased significantly in\\u0026nbsp;other doses of VB compared to control group. Dose-related relationship was recorded. Additionally, the highest DNA damage of class 3 (tail longer than 2\\u0026times; the diameter of the nucleus) was showed in mice exposed to\\u0026nbsp;10 mg/kg VB.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eMorphological sperm abnormalities\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe results in Table 5 demonstrated a significant (p˂ 0.05) and dose-dependent increase in the frequency of sperm abnormalities in comparison with the negative control. The maximum percentage of sperm defects reached 11.06% after treatment with the dose of 6 mg/kg \\u003cem\\u003evs\\u0026nbsp;\\u003c/em\\u003e3.04% for the control. With respect to the types of sperm abnormalities, Coiled tail sperm represents the major percentage. Shapeless head sperm and head without hook were also recorded. In the highest tested dose, sperm tail defects represent 7.66 % while the head abnormalities represent 3.4% of the total counted sperm.\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eChemotherapy represents the main therapeutic choice in treating cancer. The majority of chemotherapies have no selectivity to cancer cells and affect healthy body tissues where the cells are constantly growing and dividing. This fact can be explained by the side effects associated with the course of chemotherapeutic regimens. Useful drugs without side effects do not yet exist, so it is of a great importance to estimate the side effects of chemotherapy and the data must be available for patients, doctors and the pharmaceutical companies to reach the safe use of chemotherapy. However, in cancer survivors, a second malignancy frequently occurs after chemotherapy, which warrants detailed genotoxicity testing of the chemotherapeutic agents [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. Herein, a comprehensive study was performed to evaluate the genotoxic effects of vinblastine (VB) using several endpoints as biomarkers: Chromosomal aberrations in bone marrow and mice spermatocytes, DNA fragmentation in somatic and germ cells and sperm abnormalities. VB is one of the Vinca alkaloid family which represents antitumor agents from plants. From Vinca alkaloids, VB and vincristine (VC) have been well established in the treatment of cancer. The results showed that VB induced a significant percentage of chromosomal aberrations in somatic cells (bone marrow) which was dose-dependent in both single and repeated treatments. The frequency of chromosomal aberrations reached about 4-folds increase after treatment with a single dose of 10 mg/kg and about 6-folds increase after 3 successive doses at 6 mg/kg of VB as compared with the negative control. A pronounced number of polyploidy was recorded at all doses and also a number of metaphases with fragments (numerical and structural aberrations). Vinca alkaloids appear to exert their antitumor effect by binding to tubulin proteins within the cells. Because these proteins are essential contractile proteins of the mitotic spindle of dividing cells, this binding leads to mitotic arrest [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e] which may explain the appearance of a high percentage of polyploidy after VB treatment. Generally, in the absence of an intact mitotic spindle, the chromosomes may clump in an unusual grouping or may disperse throughout the cytoplasm [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. Previous studies with somatic cell cultures revealed the presence of aneuploidy and polyploidy after VB treatment [\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. Due to their action as mitotic spindle poisons, vinca alkaloids are regarded as aneugens [\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e]. Vinca alkaloids are also cell cycle phase-specific \\u0026acute;S and M\\u0026acute; phase poisons. The present study also demonstrated DNA damage in spleen cells which also recorded a dose-dependent relationship. The effect of VB in somatic cells is coincided well with the previous results of other authors: micronuclei and chromosomal aberrations were significantly induced in mouse bone marrow after treatment with VB [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e] and in cultured human lymphocytes after VC treatment [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. However, the induction of micronuclei may reflect chromosomal damage or loss of whole chromosomes from the daughter nuclei. Oxidative DNA damage in cultured human lymphocytes and an increase in the cells mitotic index (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05) were also demonstrated after VB and VC treatment [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. They also induced chromosomal mutations in \\u003cem\\u003ein vivo\\u003c/em\\u003e and in cultured cancer cells [\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e]. Moreover, VB is found to affect nucleic acids, amino acids, proteins and purine synthesis [\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e]. Tyagi et al. [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e] made a spectroscopic study showed that plant alkaloid binds with DNA \\u003cem\\u003evia\\u003c/em\\u003e A-T and G-C base pairs along with the phosphate back bone of the helix. In addition, the docking studies have indicated the interaction of VB with the adenine base of DNA helix \\u003cem\\u003evia\\u003c/em\\u003e hydrogen bonding. Conversely, other studies showed lack of mutagenic activity of Vinca alkaloids [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e]. Generally, aneugenic agents are expected to have a low level of clastogencity as a result of their poisoning effect on the mitotic spindle [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eChromosomal aberration analysis was also examined in mouse spermatocytes. The current results indicated a significant percentage of chromosomal aberrations (p ˂ 0.05) after single and repeated treatments with the presence of dose-relationship. The maximum percentage of aberrations reached approximately 3.5 and 3.2-folds increase as compared to the negative control after single dose of 10 mg/kg and repeated doses of 6 mg/kg VB respectively. The majority of aberrations were X-Y and Autosomal univalent (gonosomal univalency) and fragments. Induction of significant percentages of aberrant primary spermatocytes (P\\u0026thinsp;\\u0026le;\\u0026thinsp;0.01) was also recorded by Palo et al. [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e] with atypical bivalents. Moreover, the current results indicated DNA damage in the testes as measured by alkaline comet assay and evidenced by comet tail length. Such effect may be as a result of oxidative stress exerted by VB in the form of reactive oxygen and nitrogen species (ROS/RNS) and to its depleted activity on the antioxidant enzymes: SOD, CAT and GPx. VB was also reported to induce a disturbance in the intracellular mediator\\u0026rsquo;s \\u0026acute;H\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003e and Ca\\u003csup\\u003e2+\\u003c/sup\\u003elevel\\u0026acute; [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e]. In addition, it was demonstrated to alter calcium homeostasis \\u003cem\\u003evia\\u003c/em\\u003e mitochondrial membranes leading to cytotoxicity and chromosome instability. Inducing of apoptosis is considered one of its mechanisms of action. The present results are also concerned with measuring the morphological sperm abnormalities after VB treatment (at different doses). Different categories of abnormal sperm were noticed and indicated the transmission of VB-induced cytogenotoxic effects from spermatocyte to sperm. Jagetia et al. [\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e] demonstrated that different doses of VB can affect mouse spermatogenesis as measured by DNA flow cytometry. Previous reports also showed significant percentages of aberrant spermatogonial metaphases and chromosomal aberrations in VB-treated mice. VB also significantly increased the percentages of aberrant primary spermatocytes and the morphological sperm defects as indicated by Palo et al. [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e]. This result in addition to the results of the current work emphasizes the ability of VB to cause genetic defects that may be passed on to future generations. The cancer survivors who are treated with chemotherapy before or during their reproductive years have faced serious problem. Such effect must be taken into consideration when choosing the chemotherapeutic regimens which may jeopardize the genetic health of offspring [\\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e]. Infertility is another risk problem that faces cancer survivors. Therapeutic modalities such as radiation therapy and chemotherapy are highly effective in treating cancer, but their gonadotoxic side effects can severely impair fertility in an agent- and dose-dependent way. Spermatogenesis for example in long-term cancer survivors has elucidated evidence of persistent azoospermia or severe oligozoospermia in up to 24% of patients [\\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e]. This effect warrants several years after chemotherapy to recover. The alkylating agents such as cyclophosphamide and isophosphamide caused permanent azoospermia in 80\\u0026ndash;90% of cases [\\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e39\\u003c/span\\u003e]. Azoospermia was detected follow the use of VB in testicular cancer patients and it may be reversible within 2\\u0026ndash;3 years [\\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e40\\u003c/span\\u003e]. In the present study VB induced a high percentage of sperm defects (tail and head defects) with a pronounced appearance of coiled tail sperm. Also the absence of acrosome and amorphous or misshape head sperm was recorded. These defects might affect the ability of the sperm to reach and penetrate an egg. Coiling of the sperm tail may limit or stop its motility and also acrosome plays a crucial role in its function. Sperm motility and acrosome reaction are considered key functions in the control of reproduction and also they are essential for spermatozoa to become fertile [\\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e41\\u003c/span\\u003e]. The sperm morphology was reported to be genetically controlled by numerous autosomal and sex-linked genes [\\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e] and the formation of a normal sperm head involves intricate synchronous morphological and biochemical steps [\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e]. Oxidative stress, high levels of lipid peroxidation and oxidative DNA damage accompanied by mutation were considered the major causes of male infertility. Mutations that influence sperm quality include conditions that affect the morphological appearance of spermatozoa and their competence of fertilization [\\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e44\\u003c/span\\u003e]. The current results emphasize the positive correlation between cytogenetic damage in germ cells and sperm abnormalities.\\u003c/p\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eThe present work demonstrated that vinblastine has a genotoxic effect evidenced by DNA damage and chromosomal aberrations (CAs) induced in both somatic and germ cells of male mice. In bone marrow cells CA analysis revealed a pronounced number of polyploidy metaphases which reflect the effect of VB in the mitotic spindle. CAs induced by VB in spermatocytes were transmitted to sperm causing several defect categories. Such defects are expected to affect the fertility of the cancer survivors or might be result in malformation in the future offspring. The results of the present work must be taken into consideration while using VB in the chemotherapeutic regimens.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis work is a part of the in house project of National Research Centre (NRC), Cairo, Egypt which is under the NO: 12060167. NRC provided all necessary facilities to complete this work. The authors would like to declare that the financer had no role in the idea, practical work, discussion and publication of the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of interest:\\u0026nbsp;\\u003c/strong\\u003eThe authors declare that they have no competing interests.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthical approval\\u003c/strong\\u003e: All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All experimental procedures were carried out following the guiding principles for the care and use of laboratory animals approved by the NRC, Dokki-Giza (Egypt). The Approval Certificate is under the number: 19-163.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n \\u003cli\\u003eFacchini PJ, De Luca V (2008) Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants. The Plant Journal 54: 763-784. https://doi.org/10.1111/j.1365-313X.2008.03438.x\\u003c/li\\u003e\\n \\u003cli\\u003eMhaidat N, Alzoubi K, Khabour O, Alawneh K, Raffee L, Alsatari E, Hussein E, Bani-Hani K (2016) Assessment of genotoxicity of vincristine, vinblastine and vinorelbine in human cultured lymphocytes: a comparative study. Balkan Journal of Medical Genetics 19: 13-20. https://doi.org/10.1515/bjmg-2016-0002\\u003c/li\\u003e\\n \\u003cli\\u003eEslami A, Mathur AD, Jha KK, Wang H (2018) Acute liver failure secondary to ABVD use. Case Reports . https://doi.org/10.1136/\\u003cem\\u003ebcr\\u003c/em\\u003e-\\u003cem\\u003e2018\\u003c/em\\u003e-\\u003cem\\u003e225474\\u003c/em\\u003e\\u003c/li\\u003e\\n \\u003cli\\u003eMoudi M, Go R, Yien CYS, Nazre M (2013) Vinca alkaloids. International Journal of Preventive Medicine 4: 1231- 1235.\\u003c/li\\u003e\\n \\u003cli\\u003eChoudhury RC, Palo AK, Padhy A (2004) Cytogenetic consequences of vinblastine treatment in mouse bone marrow. Chemotherapy 50 : 171-177. https://doi.org/10.1159/000080690\\u003c/li\\u003e\\n \\u003cli\\u003eDuffin J (2000) Poisoning the spindle: serendipity and discovery of the anti-tumor properties of the vinca alkaloids. Canadian Bulletin of Medical History 17: 155-19. https://doi.org/10.3138/cbmh.17.1.155\\u003c/li\\u003e\\n \\u003cli\\u003evan Leeuwen IM, Rao B, Sachweh MC, La\\u0026iacute; S (2012) An evaluation of small-molecule p53 activators as chemoprotectants ameliorating adverse effects of anticancer drugs in normal cells. Cell Cycle 11: 1851-1861. https://doi.org/10.4161/cc.20254\\u003c/li\\u003e\\n \\u003cli\\u003ePalo AK, Pandit RS, Choudhury CR (2011) Vinblastine-induced cytogenotoxicity in spermatogonia and its transmission in the germline cells of Swiss mice. J Environ Pathol Toxicol Oncol Actions 30: 113-21. https://doi.org/10.1615/jenvironpatholtoxicoloncol.v30.i2.30\\u003c/li\\u003e\\n \\u003cli\\u003eRibatti D, Guidolin D, Conconi MT, Nico B, Baiguera S, Parnigotto PP, Vacca A, Nussdorfer GG (2003) Vinblastine inhibits the angiogenic response induced by adrenomedullin in vitro and in vivo. Oncogene 22: 6458-6461. https://doi.org/10.1038/sj.onc.1206789\\u003c/li\\u003e\\n \\u003cli\\u003eFahmy MA, Abd-Alla HI, Hassan EE, Hassan ZM, Heba-tollah MS (2020) Genotoxicity and sperm defects induced by 5-FU in male mice and the possible protective role of Pentas lanceolata-iridoids. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 503145: 850\\u0026ndash;851. https://doi.org/10.1016/j.mrgentox.2020.503145\\u003c/li\\u003e\\n \\u003cli\\u003eDiab KA, Fahmy MA, Hassan ZM, Hassan EM, Salama AB, Omara EA (2018) Genotoxicity of carbon tetrachloride and the protective role of essential oil of \\u003cem\\u003eSalvia officinalis\\u003c/em\\u003e L. in mice using chromosomal aberration, micronuclei formation, and comet assay. Environ Sci Pollut Res Int 25: 1621-1636. https://doi.org/10.1007/s11356-017-0601-2\\u003c/li\\u003e\\n \\u003cli\\u003eEvans E, Breckon G, Ford C (1964) An air-drying method for meiotic preparations from mammalian testes. Cytogenetics 3: 289-294. https://doi.org/10.1159/000129818\\u003c/li\\u003e\\n \\u003cli\\u003eHassan NH, Fahmy MA, Farghaly AA, Hassan EE (2006) Antimutagenic effect of selenium and vitamins against the genotoxicity induced by cobalt chloride in mice. 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New York, Plenum Press. https://doi.org/10.1073/pnas.72.11.4425\\u003c/li\\u003e\\n \\u003cli\\u003eFahmy MA, Hassan NHA, El-Fiky SA, Elalfy HG (2015) A mixture of honey bee products ameliorates the genotoxic side effects of cyclophosphamide. Asian Pacific Journal of Tropical Disease 5: 638-644.\\u003c/li\\u003e\\n \\u003cli\\u003eChaudhari RM, Gavit C (2015) Effect of Vincristine on some biochemical parameters in male Albino rat. Int J of Life Sciences A3: 72-76.\\u003c/li\\u003e\\n \\u003cli\\u003eLiang JC, Satya-Prakash K (1985) Induction of aneuploidy by mitotic arrestants in mouse bone marrow. Mutation Research/Genetic Toxicology 155: 61-70. https://doi.org/10.1016/0165-1218(85)90026-6\\u003c/li\\u003e\\n \\u003cli\\u003eManca A, Bassani B, Russo A, Pacchierotti F (1990) Origin of aneuploidy in relation to disturbances of cell-cycle progression. I. Effects of vinblastine on mouse bone marrow cells. 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Jin L, Lou J, He J (2008) Studying the genotoxicity of vincristine on human lymphocytes using comet assay, micronucleus assay and TCR gene mutation test in vitro. Toxicology 252: 113-117. https://doi.org/10.1016/j.tox.2008.07.057\\u003c/li\\u003e\\n \\u003cli\\u003eYamada T, Odawara K, Kaneko H (2000) Concurrent detection of gene mutations and chromosome aberrations induced by five chemicals in a CHL/IU cell line incorporating a gpt shuttle vector. 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Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 287: 29-46. https://doi.org/10.1016/0027-5107(93)90143-4\\u003c/li\\u003e\\n \\u003cli\\u003eRtibi K, Grami D, Selmi S, Amri M, Sebai H, Marzouki L (2017) Vinblastine, an anticancer drug, causes constipation and oxidative stress as well as others disruptions in intestinal tract in rat. Toxicology Reports 4: 221-225. https://doi.org/10.1016/j.toxrep.2017.04.006\\u003c/li\\u003e\\n \\u003cli\\u003eJagetia GC, Krishnamurthy H, Jyothi P (1996) Evaluation of cytotoxic effects of different doses of vinblastine on mouse spermatogenesis by flow cytometry. Toxicology 112: 227-236. https://doi.org/10.1016/0300-483X(96)03402-6\\u003c/li\\u003e\\n \\u003cli\\u003eWyrobek AJ, Schmid TE, Marchetti F (2005) Relative susceptibilities of male germ cells to genetic defects induced by cancer chemotherapies. 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Fertility and Sterility 54: 493-496. https://doi.org/10.1016/s0015-0282(16)53768-6\\u003c/li\\u003e\\n \\u003cli\\u003eVisconti P, Westbrook V, Chertihin O, Demarco I, Sleight S, Diekman A (2002) Novel signaling pathways involved in sperm acquisition of fertilizing capacity. Journal of Reproductive Immunology 53: 133-150. https://doi.org/10.1016/s0165-0378(01)00103-6\\u003c/li\\u003e\\n \\u003cli\\u003eKrzanowska H (1976) Inheritance of sperm head abnormality types in mice\\u0026ndash;the role of the Y chromosome. Genetics Research 28: 189-198. https://doi.org/10.1017/S0016672300016864\\u003c/li\\u003e\\n \\u003cli\\u003eRattner J (1972) Nuclear shaping in marsupial spermatids. J Ultrastruct Res 40: 498-512. https://doi.org/10.1016/s0022-5320(72)80038-8\\u003c/li\\u003e\\n \\u003cli\\u003eAitken RJ, Baker MA (2020) The role of genetics and oxidative stress in the etiology of male infertility-A unifying hypothesis? Front. Endocrinol 11: 581838. https://doi.org/10.3389/fendo.2020.581838\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"},{\"header\":\"Tables\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003eTable1.\\u0026nbsp;Frequency of chromosomal aberrations induced in mouse bone marrow cells after treatment with vinblastine.\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellpadding=\\\"0\\\" cellspacing=\\\"0\\\" width=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"2\\\" style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTreatment and doses\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"2\\\" style=\\\"width: 23.5266%;\\\" valign=\\\"top\\\" width=\\\"16.38830897703549%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTotal abnormal metaphases\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"3\\\" style=\\\"width: 42.9678%;\\\" valign=\\\"top\\\" width=\\\"29.958246346555324%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eNo and (%) of metaphases with different types of chromosome aberrations\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"5.614973262032086%\\\"\\u003e\\n \\u003cp\\u003eNo.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"15.37433155080214%\\\"\\u003e\\n \\u003cp\\u003eMean(%) \\u0026plusmn; SE\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"8.823529411764707%\\\"\\u003e\\n \\u003cp\\u003eGap\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"16.0427807486631%\\\"\\u003e\\n \\u003cp\\u003eFragment \\u0026nbsp; \\u0026nbsp; and/or Break\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"13.502673796791443%\\\"\\u003e\\n \\u003cp\\u003ePolyploidy\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003eI. \\u0026nbsp;Control (Non- treated)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e3.20 \\u0026plusmn; 0.37\\u003csup\\u003e\\u0026nbsp;a\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e7(1.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e9(1.80)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003eII. Vinblastine\\u003c/p\\u003e\\n \\u003cp\\u003ea-single dose\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e3 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e32\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e6.40 \\u0026plusmn; 0.60\\u003csup\\u003e\\u0026nbsp;b\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e6(1.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e10(2.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e16(3.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e4.5 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e41\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e8.20 \\u0026plusmn; 0.58\\u003csup\\u003eb,c\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e5(1.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e12(2.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e24(4.80)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e6 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e51\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e10.20 \\u0026plusmn; 0.37\\u003csup\\u003ec,d\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e6(1.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e20(4.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e25(5.0)\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e10 mg /kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e60\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e12.0 \\u0026plusmn; 0.95\\u003csup\\u003ed,e\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e8(1.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e13(2.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e39(7.80)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003eb- 3 successive dose treatment\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e3 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e65\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e13.0\\u0026plusmn; 0.95\\u003csup\\u003e\\u0026nbsp;e\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e8(1.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e12(2.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e45(9.0)\\u003c/p\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e4.5 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e83\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e16.60 \\u0026plusmn; 0.93\\u003csup\\u003e\\u0026nbsp;f\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e11(2.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e22(4.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e50(10.0)\\u003c/p\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 24.5128%;\\\" valign=\\\"top\\\" width=\\\"21.920668058455114%\\\"\\u003e\\n \\u003cp\\u003e6 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 6.3395%;\\\" valign=\\\"top\\\" width=\\\"4.384133611691023%\\\"\\u003e\\n \\u003cp\\u003e97\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.1871%;\\\" valign=\\\"top\\\" width=\\\"12.004175365344468%\\\"\\u003e\\n \\u003cp\\u003e19.40 \\u0026plusmn; 0.56\\u003csup\\u003e\\u0026nbsp;g\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 9.8615%;\\\" valign=\\\"top\\\" width=\\\"6.8893528183716075%\\\"\\u003e\\n \\u003cp\\u003e14(2.80)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 17.8915%;\\\" valign=\\\"top\\\" width=\\\"12.526096033402922%\\\"\\u003e\\n \\u003cp\\u003e25(5.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 15.074%;\\\" valign=\\\"top\\\" width=\\\"10.542797494780793%\\\"\\u003e\\n \\u003cp\\u003e58(11.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eA total of 500 cells were analyzed (5 mice per group; 100 cells/mouse). One way ANOVA\\u0026ndash;Tukey\\u0026rsquo;s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Table2. DNA fragmentation detected in spleen tissues of mice exposed to different doses vinblastine.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellpadding=\\\"0\\\" cellspacing=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTreatment and doses\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003eDNA Fragmentation %\\u003c/p\\u003e\\n \\u003cp\\u003e(M \\u0026nbsp; \\u0026nbsp; \\u0026plusmn; SEM)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eChange\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eInhibition %\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003e(-ve) control\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003e9.5\\u0026plusmn;0.71\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e0.00\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e0.00\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003eVinblastine\\u003c/p\\u003e\\n \\u003cp\\u003e3 mg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003e10.2\\u0026plusmn;0.49\\u003csup\\u003eab\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e0.70\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e95.24\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003e4.5 mg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003e12.4\\u0026plusmn;0.47\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e2.90\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e80.27\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003e6 mg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003e18.6\\u0026plusmn;0.67\\u003csup\\u003edc\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e9.10\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e38.09\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"31.987577639751553%\\\"\\u003e\\n \\u003cp\\u003e10 mg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"27.950310559006212%\\\"\\u003e\\n \\u003cp\\u003e24.2\\u0026plusmn;0.88\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"16.770186335403725%\\\"\\u003e\\n \\u003cp\\u003e14.7\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"23.29192546583851%\\\"\\u003e\\n \\u003cp\\u003e29.28\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eMeans with different superscripts (\\u003csup\\u003ea, b,c,d\\u003c/sup\\u003e) between groups in the same column are significantly different at P\\u0026lt;0.05.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eTable3.\\u0026nbsp;Frequency of chromosomal aberrations induced in mouse spermatocytes after treatment with vinblastine.\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellpadding=\\\"0\\\" cellspacing=\\\"0\\\" width=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"2\\\" width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTreatment and doses\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"2\\\" width=\\\"20.689655172413794%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTotal abnormal metaphases\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"6\\\" width=\\\"56.03448275862069%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eNo and (%) of metaphases with different types of chromosome aberration\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"7.865168539325842%\\\"\\u003e\\n \\u003cp\\u003eNo.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.10112359550562%\\\"\\u003e\\n \\u003cp\\u003eMean(%) \\u0026plusmn; SE\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.48314606741573%\\\"\\u003e\\n \\u003cp\\u003eX-Y univalent\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.606741573033707%\\\"\\u003e\\n \\u003cp\\u003eAutosomal univalent\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.112359550561798%\\\"\\u003e\\n \\u003cp\\u003eX-y u.\\u003c/p\\u003e\\n \\u003cp\\u003e+\\u003c/p\\u003e\\n \\u003cp\\u003eA-u.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"13.48314606741573%\\\"\\u003e\\n \\u003cp\\u003eFragment \\u0026nbsp; \\u0026nbsp; and/or Break\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.112359550561798%\\\"\\u003e\\n \\u003cp\\u003eChain IV\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.235955056179776%\\\"\\u003e\\n \\u003cp\\u003ePolyp.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003eI. \\u0026nbsp;Control (Non-treated)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e19\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e3.80 \\u0026plusmn; 0.37\\u003csup\\u003e\\u0026nbsp;a\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e17(3.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e2(0.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003eII. \\u0026nbsp; Vinblastine\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp; \\u0026nbsp; \\u0026nbsp;a- Single dose\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e3 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e40\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e8.0 \\u0026plusmn; 0.77\\u003csup\\u003e\\u0026nbsp;b\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e25(5.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e12(2.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e1(0.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e2(0.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e4.5 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e44\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e8.80 \\u0026plusmn; 0.37\\u003csup\\u003eb,c\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e28(5.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e12(2.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e1(0.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e3(0.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e6 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e52\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e10.40 \\u0026plusmn; 0.93\\u003csup\\u003ec,d\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e23(4.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e20(4.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e1(0.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e3(0.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e1(0.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e4(0.80)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e10 mg /kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e66\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e13.20 \\u0026plusmn; 1.16\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e45(9.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e17(3.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e2(0.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e2(0.20)\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003eb - 3 successive dose treatment\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp; \\u0026nbsp;3 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e51\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e10.20 \\u0026plusmn; 0.86\\u003csup\\u003eb,c,d\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e27(5.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e18(3.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e4(0.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e1(0.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e1(1.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e4.5 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e55\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e11.0 \\u0026plusmn; 0.83\\u003csup\\u003ec,d,e\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e41(8.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e8(1.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e5(1.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e1(1.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"23.275862068965516%\\\"\\u003e\\n \\u003cp\\u003e6 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"6.0344827586206895%\\\"\\u003e\\n \\u003cp\\u003e61\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"14.655172413793103%\\\"\\u003e\\n \\u003cp\\u003e12.20 \\u0026plusmn; 0.37\\u003csup\\u003ed,e\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e30(6.0)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.206896551724139%\\\"\\u003e\\n \\u003cp\\u003e18(3.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10.344827586206897%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e6(1.20)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"7.758620689655173%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"8.620689655172415%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e7(1.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eA total of 500 cells were analyzed (5 mice per group; 100 cells/mouse). X-Y u: X-Y univalent, A-u: Autosomal univalent, Polyp: Polyploidy. One way ANOVA\\u0026ndash;Tukey\\u0026rsquo;s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Table 4. Visual score of DNA damage in the testes of male mice exposed to different doses of vinblastine.\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellpadding=\\\"0\\\" cellspacing=\\\"0\\\" width=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"2\\\" valign=\\\"top\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003eTreatment\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd rowspan=\\\"2\\\" valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eNo of samples\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"2\\\" valign=\\\"top\\\" width=\\\"21.818181818181817%\\\"\\u003e\\n \\u003cp\\u003eNo. of cells\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"4\\\" valign=\\\"top\\\" width=\\\"22.727272727272727%\\\"\\u003e\\n \\u003cp\\u003eClass\\u003csup\\u003e**\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd rowspan=\\\"2\\\" valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003eDNA damaged cells %\\u003c/p\\u003e\\n \\u003cp\\u003e(Mean \\u0026plusmn; SE)\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"26.53061224489796%\\\"\\u003e\\n \\u003cp\\u003eAnalyzed\\u003csup\\u003e*\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"22.448979591836736%\\\"\\u003e\\n \\u003cp\\u003eComets\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"14.285714285714286%\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.244897959183673%\\\"\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.244897959183673%\\\"\\u003e\\n \\u003cp\\u003e2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.244897959183673%\\\"\\u003e\\n \\u003cp\\u003e3\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003eI- (-ve) control\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.818181818181818%\\\"\\u003e\\n \\u003cp\\u003e500\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e36\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e464\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e11\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e7.21\\u0026plusmn;1.07\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003eII- Vinblastine\\u003c/p\\u003e\\n \\u003cp\\u003e3\\u0026nbsp;mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.818181818181818%\\\"\\u003e\\n \\u003cp\\u003e500\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e39\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e461\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e23\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e7.83\\u0026plusmn;0.93\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003e4.5\\u0026nbsp;mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.818181818181818%\\\"\\u003e\\n \\u003cp\\u003e500\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e51\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e449\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e28\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e17\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e6\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e10.24\\u0026plusmn;1.24\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003e6\\u0026nbsp;mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.818181818181818%\\\"\\u003e\\n \\u003cp\\u003e500\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e83\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e417\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e31\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e28\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e24\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e16.61\\u0026plusmn;1.50 \\u003csup\\u003ecd\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" width=\\\"24.545454545454547%\\\"\\u003e\\n \\u003cp\\u003e10\\u0026nbsp;mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.818181818181818%\\\"\\u003e\\n \\u003cp\\u003e500\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e107\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e393\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e35\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e31\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e41\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e21.43\\u0026plusmn;1.21\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003e*: Number of cells examined per a group, \\u003csup\\u003e**\\u003c/sup\\u003e: Class 0= no tail; 1= tail length \\u0026lt; diameter of nucleus; 2= tail length between 1\\u0026times;, and 2\\u0026times; the diameter of nucleus; and 3= tail length \\u0026gt; 2\\u0026times; the diameter of nucleus. Data are presented as Mean \\u0026plusmn; SE. Mean values within tissue with unlike superscript letters were significantly different (\\u003cem\\u003eP\\u003c/em\\u003e\\u0026lt;0.05).\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eTable5.\\u0026nbsp;Percentage of sperm abnormalities induced in male mice after treatment with vinblastine\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellpadding=\\\"0\\\" cellspacing=\\\"0\\\" width=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"4\\\" valign=\\\"top\\\" width=\\\"16.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eExperimental groups and doses\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"2\\\" rowspan=\\\"3\\\" valign=\\\"top\\\" width=\\\"20.90909090909091%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTotal abnormal sperm\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"5\\\" valign=\\\"top\\\" width=\\\"62.72727272727273%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eNo. and (%)of different types of sperm abnormalities\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd colspan=\\\"4\\\" rowspan=\\\"2\\\" valign=\\\"top\\\" width=\\\"68.1159420289855%\\\"\\u003e\\n \\u003cp\\u003eHead abnormalities\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"31.884057971014492%\\\"\\u003e\\n \\u003cp\\u003eTail abnormalities\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"100%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"6.521739130434782%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eNo.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"18.47826086956522%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eMean (%)\\u0026plusmn;SE\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"15.217391304347826%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eAmorphous\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.956521739130435%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eWithout hook\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.956521739130435%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eTriangle\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"11.956521739130435%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eBanana\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"23.91304347826087%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eCoiled tail\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"16.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eI- Control (-ve)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e152\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"15.454545454545455%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e3.04 \\u0026plusmn; 0.19\\u003csup\\u003e\\u0026nbsp;a\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.727272727272727%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e16(0.32)\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e4(0.08)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e2(0.04)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e1(0.02)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e20(0.40)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"16.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003eII- Vinblastine\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e3 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e472\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"15.454545454545455%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e9.44 \\u0026plusmn;0.13\\u003csup\\u003e\\u0026nbsp;b\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"12.727272727272727%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e71(1.42)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e69(1.38)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e18(0.36)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e23(0.46)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e291(5.82)\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"16.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e4.5 mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e520\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"15.454545454545455%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e10.40 \\u0026plusmn; 0.73\\u003csup\\u003eb,c\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.727272727272727%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e85(1.70)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e67(1.34)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e19(0.38)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e19(0.38)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e330(6.60)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"16.363636363636363%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e6mg/kg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"5.454545454545454%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e553\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"15.454545454545455%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e11.06\\u0026plusmn;0.14\\u003csup\\u003e\\u0026nbsp;c\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"12.727272727272727%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e54(1.08)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e87(1.74)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e18(0.36)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"10%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e11(0.22)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" width=\\\"20%\\\"\\u003e\\n \\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n \\u003cp\\u003e383(7.66)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eTotal number of examined sperms 5000 per each treatment (1000 /mouse, 5 mice/group) .One way ANOVA\\u0026ndash;Tukey\\u0026rsquo;s multiple comparisons test was used. The values having different superscript letters in each column are significantly different from one another at p\\u0026lt;0.05.\\u003c/p\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":true,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"molecular-biology-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"mole\",\"sideBox\":\"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)\",\"snPcode\":\"11033\",\"submissionUrl\":\"https://submission.nature.com/new-submission/11033/3\",\"title\":\"Molecular Biology Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"Vinblastine, Mitotic inhibitor, Chromosomal aberrations, DNA damage, Sperm abnormalities\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-1943872/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-1943872/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"Background\\n\\nGenotoxicity studies of chemotherapeutic drugs is of special need. Secondary tumors may develop many years after treatment as a result of chemo genotoxicity. The effect of chemo on meiotic chromosomes and sperm defects is another complication associated with chemo treatment. In this study the genotoxicity of vinblastine (VB) was estimated in both somatic and germ cells.\\n\\nMaterials\\n\\n85 mice were taken. 4 single doses of VB at 3, 4.5, 6 and 10 mg/kg and 3 successive doses at 3, 4.5 and 6 mg/kg were taken for estimation of chromosomal aberrations (CAs). 4 single doses of VB were involved in estimating the DNA fragmentation, and comet assay. Samples were taken 24 h after the last treatment. For sperm abnormalities mice were injected with 3 successive doses of VB at 3, 4.5, and 6 mg/kg and samples were taken 35 days after the 1st injection.\\n\\nResults\\n\\nThe results demonstrated a significant frequency of DNA fragmentation in spleen cells and in the percentage of CAs in bone marrow. Numerical and structural aberrations were recorded with a pronounced number of polyploidy metaphases. VB also induced a significant percentage of CAs in spermatocytes in the form of univalent. Sperm defects in the form of coiled tail, absence of acrosome and shapeless head and a significant DNA damage in the testes were recorded.\\n\\nConclusion\\n\\nVB is genotoxic in somatic and germ cells. Sperm defects induced by VB are of serious concern to future generations and may affect the fertility of cancer survivors.\",\"manuscriptTitle\":\"Genotoxicity, DNA damage and sperm defects induced by vinblastine\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2022-08-17 20:36:24\",\"doi\":\"10.21203/rs.3.rs-1943872/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewerAgreed\",\"content\":\"\",\"date\":\"2022-08-12T13:04:46+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2022-08-12T11:45:57+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2022-08-09T15:17:10+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Molecular Biology Reports\",\"date\":\"2022-08-09T03:01:49+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"molecular-biology-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"mole\",\"sideBox\":\"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)\",\"snPcode\":\"11033\",\"submissionUrl\":\"https://submission.nature.com/new-submission/11033/3\",\"title\":\"Molecular Biology Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"50afe7e5-6ffa-4e59-bbd4-c1b8d5eeaf5d\",\"owner\":[],\"postedDate\":\"August 17th, 2022\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2022-10-27T07:41:39+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2022-08-17 20:36:24\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-1943872\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-1943872\",\"identity\":\"rs-1943872\",\"version\":[\"v1\"]},\"buildId\":\"WrCJVZZCHTDjtuVLN7oU0\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}