Significant Differences In Mitotic Index Research Articles

Higher toxicity of nano-scale TiO2 and dose-dependent genotoxicity of nano-scale SiO2 on the cytology and seedling development of broad bean Vicia faba

Since nanotechnology entered the field of agriculture, its safety impact on crops has been a high priority interest. Here, we aimed to evaluate the effect of two different types of nanoparticles (n-), n-SiO2 and n-TiO2, on the above- and below-ground growth and the root-tip cell mitosis of broad beans (Vicia faba L.), one of the major carbohydrate food sources as well as an ecotoxicological model plant. Seeds were soaked in n-SiO2 and n-TiO2 each at different concentrations (25, 50 and 75 mg/L) for 24 h. Nano-TiO2 decreased vigor index, reflecting shorter shoots at all concentrations studied. By contrast, germination percentage and root length were not affected by any treatments. Cytological analysis suggested no significant difference in mitotic index (index for cell division activity) from the control. However, total chromosomal aberrations (%) were increased dose-dependently by n-SiO2 and dose-independently by n-TiO2. Also, different types of chromosomal abnormalities were induced by the nanomaterials; n-SiO2 induced bridges at 50 and 75 mg/L, whereas n-TiO2 induced breaks at 50 mg/L. In addition, cells in prophase were more frequently observed and those in anaphase less frequently seen with decreasing n-SiO2 concentrations. We concluded that n-TiO2 was more toxic than n-SiO2 for broad bean chromosomes and early plant development at the concentrations studied. Finally, our review indicates the lack of evidence of germination enhancement by n-TiO2 in Poaceae, a large monocotyledon family, which may require further attention.

THE CYTOPROTECTIVE ACTIVITY OF AQUEOUS GREEN TEA EXTRACT AGAINST METRONIDAZOLE AND TINIDAZOLE GENOTOXIC EFFECT

Objective: The study was designed to evaluate the genoprotective effect of green tea extract against genotoxicity induced by metronidazole and tinidazole.Methods: A total of 36 mice were used, for each experiment. The animals were allocated into six groups: Group I - negative control administered distilled water; Group II - healthy mice treated with metronidazole alone, Group III - healthy mice treated with tinidazole alone, Group IV - healthy mice administered green tea extract alone, Group V - healthy mice administered of metronidazole anddd then green tea extract was administered, and Group VI - healthy mice administered of tinidazole and then green tea extract was administered.Results: It has been found that there are significant differences in mitotic index and micronucleus (MN) appearance between the Groups II, IV, and V (7.76±0.8, 11.92±1.14, and 8.36±0.57 and 6.92±0.5, 10.18±1.19, and 7.52±1.05) in bone marrow cells and spleen cells for mitotic index, respectively (6.75±0.4, 5.5±0.41, and 5.92±0.68), in bone marrow for MN appearance. Furthermore, it has been found that there are significant differences in mitotic index and MN appearance between the Groups III, IV, and VI (4.36±0.88, 11.92±1.14, and 5.36±0.55 and 4.08±0.35, 10.18±1.19, and 5.08±0.35) in bone marrow cells and spleen cells for mitotic index, respectively (8.43±0.96, 5.5±0.41, and 6.84±0.66), in bone marrow for MN appearance.Conclusion: The aqueous extract of green tea has a protective effect against the genotoxic effects induced by either metronidazole or tinidazole in mice.

THE CYTOPROTECTIVE ACTIVITY OF AQUEOUS GREEN TEA EXTRACT AGAINST METRONIDAZOLE AND TINIDAZOLE GENOTOXIC EFFECT

Objective: The study was designed to evaluate the genoprotective effect of green tea extract against genotoxicity induced by metronidazole and tinidazole.Methods: A total of 36 mice were used, for each experiment. The animals were allocated into six groups: Group I - negative control administered distilled water; Group II - healthy mice treated with metronidazole alone, Group III - healthy mice treated with tinidazole alone, Group IV - healthy mice administered green tea extract alone, Group V - healthy mice administered of metronidazole anddd then green tea extract was administered, and Group VI - healthy mice administered of tinidazole and then green tea extract was administered.Results: It has been found that there are significant differences in mitotic index and micronucleus (MN) appearance between the Groups II, IV, and V (7.76±0.8, 11.92±1.14, and 8.36±0.57 and 6.92±0.5, 10.18±1.19, and 7.52±1.05) in bone marrow cells and spleen cells for mitotic index, respectively (6.75±0.4, 5.5±0.41, and 5.92±0.68), in bone marrow for MN appearance. Furthermore, it has been found that there are significant differences in mitotic index and MN appearance between the Groups III, IV, and VI (4.36±0.88, 11.92±1.14, and 5.36±0.55 and 4.08±0.35, 10.18±1.19, and 5.08±0.35) in bone marrow cells and spleen cells for mitotic index, respectively (8.43±0.96, 5.5±0.41, and 6.84±0.66), in bone marrow for MN appearance.Conclusion: The aqueous extract of green tea has a protective effect against the genotoxic effects induced by either metronidazole or tinidazole in mice.

Avaliação da atividade genotóxica, citotóxica e antimicrobiana da infusão das folhas de Spondias purpurea L.

O uso de plantas medicinais cada vez mais é mostrado como uma fonte alternativa para tratamento de doenças. Este trabalho visou determinar o nível genotóxico e citotóxico da infusão das folhas de Spondias purpurea L., investigar o potencial antimicrobiano e conhecer o perfil cromatográfico de compostos presentes na infusão. Foram realizados os testes Allium cepa e Artemia salina para avaliar a genotoxicidade e citotoxicidade, respectivamente. Os métodos de difusão em disco e concentração inibitória mínima (CIM) foram usados para determinar a atividade antibacteriana nas bactérias Staphylococcus epidermidis, S. aureus, Escherichia coli, Salmonella enterica sorotipo Choleraesuis, Pseudomonas aeruginosa e Klebsiella pneumoniae. No teste de A. cepa, a análise estatística não indicou diferenças significativas do índice mitótico entre os tratamentos e o controle. O teste de A. salina mostrou que a infusão apresentou toxicidade frente aos náuplios a partir da dose 10µg/mL. Em relação à atividade antibacteriana, houve presença de halos de inibição contra as bactérias S. aureus e E. coli, e quando submetidos ao CIM, a infusão apresentou atividade antibacteriana contra todas as bactérias testadas. Sendo assim, a partir desse estudo pode-se inferir que a infusão (chá) das folhas da planta S. purpurea se mostrou como uma fonte medicinal não genotóxica para o teste A. cepa, bem como uma possível fonte antitumoral indicada pelo teste A. salina, além de uma promissora fonte antibacteriana.

Evaluation of Peritoneal Macrophages Migration Inhibition and Lymphocytes Mitotic Index in Shigella Dysenteriae Type1 Immunized Guinea Pigs

Background and objective: To determine the performance and functional activity of macrophage and lymphocytes after immunization with phenol killed Shigella dysenteriae type 1Materials and Methods: phenol killed Shigella dysenteriae type 1 (16 103bacteria /ml ) was injected subcutaneously(S/C) in 10 guinea pigs at a dose of 0.5 ml .At the 3rd week animals given a 1st booster dose of (1 ml) of killed bacteria. At 5th week, a 2nd booster dose (1ml) was given (S/C) . Five animals were used as control group and injected with phosphate buffer saline . After isolation of peritoneal macrophage and Blood Lymphocytes, macrophage migration inhibition (MMIT) and lymphocytes, transformation (LTT) tests have been applied.Results: The level of inhibition of migration decreased gradually, while reducing the concentration of the PHA antigen. Migration inhibition coefficient was (0.477 0.268) when PHA antigen (400 g / ml) , (0.610 0.181) when PHA (40 g / ml), (0.641 0.145) when PHA antigen (4 g / ml). No significant difference in migration inhibition coefficient was detected between immunized groups (p value =0.118245) at different concentration of PHA antigen. The control group did not show any migration inhibition for macrophages at different concentrations of PHA antigen. Significant difference was detected between control groups (p value =0.033035) at different concentration of PHA antigen. Significant difference (p value =0.00759 ) , (p value =0.017049) , (p value =0.000278) was detected between immunized and control groups at 4 g / ml, 40 g / ml, 400 g / ml, concentration of PHA antigen respectively . Lymphocyte transformation test showed a clear response of the lymphocytes toward PHA. The mitotic index (Ml) of the lymphocytes exposed to the (0.1mg/ml) PHA antigen was(11.56 3.22 )and for (5mg/ml) PHA was (41.49 26.298 )and for control was (1.18 0.516). Significant difference in mitotic index was detected between groups (p value =0.003554) according to the dose of PHA antigen.Conclusions: MMIT and LTT can be used effectively to evaluate and monitoring of immunological responsiveness to shigella dysenteriae type 1 antigen and both MMIT and LTT proportionally associated with pre sensitization dose and duration of exposure to antigen.

Proliferative behavior of vaginal fibroblasts from women with pelvic organ prolapse

ObjectivePelvic organ prolapse (POP) significantly impacts quality of life of women, especially with advancing age. Cell proliferation is a critical parameter in both normal and pathophysiological processes. We sought to examine fibroblast proliferation in premenopausal women with and without POP and menopausal women with POP, and examine whether TGF-β1, a fibroblast mitogen, could stimulate proliferation in vaginal fibroblasts from these populations. Study designVaginal wall biopsies were obtained from asymptomatic women (controls) and women with POP (cases). Fibroblasts were cultured from these tissues. Vaginal fibroblasts were treated with or without TGF-β1. Cell proliferation rate (mitotic index) was measured with time-lapse dark-field microscopy. Cell mitosis was counted with ImageJ software after analysis of time-lapse images as Quick time movies. ResultsThere was no significant difference in mitotic index throughout different time points of observation between premenopausal controls and cases of similar ages. However, a significant difference in mitotic index was seen between premenopausal and menopausal cases (p=0.01), with the menopausal group exhibiting significantly lower mitotic indices. When treated with different doses of TGF-β1, premenopausal control fibroblast proliferation increased with 5ng/ml of TGF-β1 compared to non-treated fibroblasts (p=0.04). TGF-β1 stimulation did not affect fibroblasts from either premenopausal or menopausal cases. ConclusionsVaginal fibroblast proliferation decreases with age and this association does not appear to be affected by the presence of pelvic organ prolapse. TGF-β1 stimulation increased cell proliferation of premenopausal control fibroblasts. In contrast, there was no response seen in fibroblasts from premenopausal and menopausal cases.

Genotoxicity Studies of Radish Grown on Industrial and Domestic Waste Water Using Alium cepa Root Tip Assay

In absence of adequate water supply local vegetable vendors in Mumbai use sewage and industrial waste water for agricultural purpose. This sewage and industrial waste water contains various organic and inorganic compounds which have direct health effects on human life. The response of Alium cepa genetic material can be used to evaluate the effect of potential genotoxic and cytoxic substances in the environment. In the present study, we have used Alium cepa root tip assay to evaluate physiological and genotoxic effects of waste water on vegetables, particularly radish grown along the areas of railway track. Samples were collected from three different sites in Mumbai region and analyzed for their heavy metal content. Similarly, effect of contaminated waste water on the external morphology of onion roots was observed using root length inhibition assay. Genotoxicity was also determined by observing the Mitotic Index (MI) and Chromosomal Aberrations (CA). Significant differences in MI and root length inhibition with increasing concentrations of different samples were observed. The diverse CA were increased significantly (p<0.05) with increase in concentration. Apart from this, bioaccumulation was also studied in soil and radish samples where concentration of heavy metals like Cd, Pb was significantly high. Consuming such radishes grown on untreated water poses health hazards to human population. Frequent and continual use of untreated waste water leads to bioaccumulation in radish as well as makes soil unfit for agricultural use.

Risk evaluation of industrial wastewater on plants using onion(Allium cepa l) chromosome aberration assay

Rapid strides in industrialization coupled with incomplete sewage treatment system have led to indiscriminate discharge of potentially mutagenic wastewater into agricultural lands and water bodies. The response of Onion plant genetic material to the presence of potential cytotoxic and genotoxic substances in the environment was used to evaluate the toxicity of chemical effluent collected from a chemical industry in Enugu, Nigeria. The Allium test procedure was used. The concentrations of heavy metals (Cu, Co, Cd, Pb) were also determined. The data obtained showed that the mitotic index (MI) decreased as effluent concentration and treatment durations increased. Significant differences in MI at different concentrations and treatment durations were observed. Conversely, percentage of abnormal dividing cells significantly (P<0.05) increased as the concentration and treatment durations increased. Diverse chromosomal aberrations and cytotoxic problems were observed. These ranged from a mild C-mitotic effect at lower concentrations to vacuolated nucleues and even cytokinetic failure at higher concentrations. The adverse effects of this untreated wastewater on the ecosystem of which one of the most vulnerable are plants are discussed.

Liver Regeneration and Hemodynamics in Pigs With Mesocaval Shunt

A surgical technique using a mesocaval shunt and downstream ligation of the superior mesenteric vein has been recently proposed to overcome the size limitations that restrict the use of partial liver grafts. We designed an experimental study in pigs to evaluate the capacities of liver regeneration and hemodynamic changes after completion of this procedure. Liver regeneration after left hepatectomy was compared between two groups of five pigs, with or without mesocaval shunt, sacrificed 11 to 14 days after surgery. A third group of five animals was used for hemodynamic studies. Liver regeneration in study animals was 45.3% of controls. This was obtained despite a reduction of the venous inflow to 15.6% of the control, resulting in a net decrease of the total blood inflow to 56% of the control, despite a compensatory increase in the arterial inflow. There was no significant difference in mitotic index, hepatocellular size, and glycogen content between study and control animals. Our experimental study confirms that the regenerative capacities of the pig liver are largely preserved despite the dramatic reduction of the venous blood inflow, reduced to its gastroduodenosplenopancreatic component. This lends further support to the hypothesis that the gastroduodenosplenopancreatic blood is enriched in hepatotrophic factors, likely to originate from the pancreas and duodenum.

Morphometric Analysis of the Epithelium of Mammary Fibroadenomas During the Proliferative and Secretory Phases of the Menstrual Cycle.

Fibroadenoma of the breast is a benign neoplasia with a mixture of glandular and mesenchymal elements of high incidence and is mainly present in young women. This aspect and the presence of estrogen and progesterone receptors reinforce its hormone dependency. Recent studies have demonstrated that the proliferative activity of the mammary epithelium is significantly increased during the luteal phase. In order to assess whether fibroadenomas behave like the normal breast under the cyclic action of hormones we counted the number of mitoses and calculated the nuclear volume of epithelial cells in both the proliferative and luteal phases. We selected 20 women with clinically diagnosed fibroadenomas, 10 in the proliferative phase and 10 in the secretory phase. Our results showed no significant differences in mitotic index or nuclear volume between the proliferative and the luteal phases of fibroadenomas. We therefore conclude that the epithelium of fibroadenomas behaves like a true neoplasia whose regulation may occur via paracrine mechanisms, which in turn would explain its self-limited growth.

Reduction in apoptosis relative to mitosis in histologically normal epithelium accompanies fibrocystic change and carcinoma of the premenopausal human breast

The aims of this study in 227 premenopausal women were (a) to determine the mitotic index (MI), the thymidine labelling index (LI), and the apoptotic index (AI) within the epithelial cells of histologically 'normal' human breast biopsy material removed away from the site of either a fibroadenoma or a carcinoma; and (b) to relate differences in the kinetic indices of the 'normal' epithelium to the pathology in the same breast diagnosed as fibroadenoma alone (125 patients), fibroadenoma with accompanying mild fibrocystic change (79 patients), or carcinoma (23 patients). Ratios of the average indices (AI/MI, AI/LI, MI/LI) were also calculated to minimize uncertainties related to the total cell population counted, the denominator in the LI, MI, and AI. All indices and ratios of indices were corrected for age, averaged over the cycle, and expressed as log-transformed values for analysis. Significant (P less than 0.001) reductions in AI and in apoptosis relative to mitosis (reduced AI/MI) were found in 'normal' epithelium from breasts having fibrocystic change (AI = 0.17 +/- 0.02; AI/MI = 1.01 +/- 0.18) and carcinoma (AI = 0.19 +/- 0.04; AI/MI = 0.88 +/- 0.29), compared with breast with fibroadenoma alone (AI = 0.27 +/- 0.03; AI/MI = 1.29 +/- 0.39). In the absence of significant differences in MI and LI between the 'normal' tissue groups, this finding raises the possibility that reduced epithelial cell apoptosis might be causally associated with the development of fibrocystic change and with an increased risk of development of carcinoma.(ABSTRACT TRUNCATED AT 250 WORDS)

Prostaglandins and the colonic epithelium: Effects of misoprostol on crypt size, cell production, and cell migration in the dog

Colonic epithelial cell division, cell migration, and cell transit were investigated in dogs given 300 μg · kg−1 · day−1 of the prostaglandin E1 analogue, misoprostol, for 11 weeks. The animals were then injected with [3H]thymidine and killed at timed intervals. The distribution of labeled and mitotic cells within the crypts was determined by scoring autoradiographs. There were no significant differences in mitotic index or labeling index between the two groups. The data were pooled and converted to give crypt cell production rates of 51.1 ± 11.1 (control) and 58.24 ± 8.6 cells per crypt per hour (test). However, the crypt length and cell population were slightly, but significantly, greater in the misoprostol-treated group (P < 0.01). The movement of the wave of labeled cells with time after injection was used to calculate the median cell migration rates, which were 23.50 ± 3.03 cell positions per day (control) and 18.30 ± 2.56 (test). Thus, misoprostol had no significant effect on either the cell migration rate or the transit rates.

Analyses for in vitro growth conditions, cytokinetics, and sister chromatid exchanges in lymphocytes cultured from the Sprague-Dawley rat.

Peripheral blood samples from Sprague-Dawley rats gave successful lymphocyte growth in GIBCO: 1A, RPMI 1640, and Eagle's minimum essential medium (MEM) culture media. Various growth conditions, cytokinetics, and sister chromatic exchange (SCE) induction were studied using reconstituted GIBCO 1A only. Neither methoxyflurane anesthesia of the rats before sampling nor washing of the cells with phosphate buffered saline affected the mitotic index. Cultures treated with [3H]thymidine showed the lymphocytes entering into DNA synthesis after approximately 24 h. The time at which BUdR (5-bromo-2' deoxyuridine) was added, i.e. 0 vs. 24 h incubation, had minimal effect on the mitotic index of cultures harvested at 48 h. However, when harvest was extended to 72 h, mitotic activity was greater in the cultures treated with BUdR at 24 h. No significant differences in mitotic index and the number of average lymphocyte division were detected in cultures exposed to 0.3 to 0.5 microgram/ml BUdR at 24 h and harvested at 72 h. Although SCE frequencies increased in the presence of BUdR, the baseline level of SCEs was estimated to be 5 to 6/cell. Average generation time of the lymphocytes dividing between 48 and 72 h was 16.5 h. Because of its simplicity of culture and the reproducible nature of its in vitro growth kinetics, the Sprague-Dawley rat lymphocyte is a suitable model for cytogenetic investigations.

Mechanism of size regulation in mouse embryo aggregates

A detailed comparison of the postimplantation development of normal and double-sized mouse embryos, produced by aggregating two 8-cell stage eggs, revealed that size regulation occurred in the double embryos between 5 days, 16 h post coitum (p.c.) and 6 days, 16 h p.c. Size regulation occurred simultaneously in all tissues, suggesting that a single regulatory mechanism may control size in the early embryo. Size regulation appeared to be brought about by alteration in cell cycle length. There was no obvious increase in cell death in the double embryos nor an increase in the non-dividing cell population. However, colcemid treatment revealed a significant difference in mitotic index between double and control embryos over the period of size regulation. Control embryos showed a proliferative burst around 6 days, 8 h p.c. which did not occur in the double embryos. It is not yet clear whether this control of proliferative activity in double embryos is exerted by the embryo itself or by the uterine environment. Histological analysis also suggested that proamniotic cavity formation, which occurs before size regulation, was dependent on total cell number and not on the number of cell cycles undergone since fertilization. Proamniotic cavity formation was observed to occur at different times but at similar cell numbers in double, control and half embryos.

Estradiol-17β- progesterone and 5α-dihydrotestosterone receptors of uterine myometrium and myoma in the human subject

Steroid receptors in human uterine myomas and the corresponding myometria were characterized and investigated to clarify their relation to tumor growth. The 248,800 g supernatant of tissue homogenates was used, in which steroid receptors were characterized and determined by Scatchard plot analysis using dextran coated charcoal at 4°C. In cytosols of both myoma and moymetrium, the estrogen receptors (ER) were sedimented at approximately the 7S, 5S and 4S regions, and the estradiol-17β(E 2)-ER complex had a dissociation constant ( K D ) = 4.5 × 10 −10M. Progesterone receptors (PR) were sedimented at approximately the 7 S and 4–5 S regions. The K D of the progesterone-PR complex was 1.5 × 10 −9M and the K D of the R5020-PR complex was 5.0 × 10 −10M. Androgen receptors (AR) were sedimented at approximately the 6–7 S and 5 S regions and dihydrotestosterone (DHT)-AR complex had a K D = 4.0 × 10 −10M. All of steroid-7 S binding in the cytosol were easily degraded during 5–20% sucrose gradient centrifugation. Ligand specificity studies characterized the specificity of receptors. There was no significant difference in steroid receptor levels (expressed as fmol/mg protein) between myoma and the corresponding myometrium; however if the receptor level was expressed as fmol/μDNA, the estrogen receptor level in the myometrium was higher than that in the myoma ( P < 0.05). There was no significant difference in mitotic index between the two tissues.