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Environmental Toxicology and Chemistry Mar 2021Per- and polyfluoroalkyl substances (PFAS) have emerged as contaminants of global concern. Among several PFAS, perfluorooctanoic acid (PFOA) and perfluorooctane...
Per- and polyfluoroalkyl substances (PFAS) have emerged as contaminants of global concern. Among several PFAS, perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) are persistent and bioaccumulative compounds. We investigated the cyto-genotoxic potential of PFOS to Allium cepa root meristem cells. The A. cepa root tips were exposed to 6 different concentrations (1-100 mg L ) of PFOS for 48 h. Reduction in mitotic index and chromosomal aberrations was measured as genotoxic endpoints in meristematic root cells. Exposure to PFOS significantly affected cell division by reducing the miotic index at higher concentrations (>10 mg L ). The median effect concentration of PFOS to elicit cytotoxicity based on the mitotic index was 43.2 mg L . Exposure to PFOS significantly increased chromosomal aberrations at concentrations >25 mg L . The common aberrations were micronuclei, vagrant cells, and multipolar anaphase. The alkaline comet assay revealed a genotoxic potential of PFOS with increased tail DNA percentage at concentrations >25 mg L . To our knowledge, this is the first study to report the cyto-genotoxic potential of PFOS in higher plants. Environ Toxicol Chem 2021;40:792-798. © 2020 SETAC.
Topics: Alkanesulfonic Acids; Chromosome Aberrations; DNA Damage; Fluorocarbons; Meristem; Mitotic Index; Onions; Plant Roots
PubMed: 33074584
DOI: 10.1002/etc.4905 -
Zhonghua Bing Li Xue Za Zhi = Chinese... May 2021To study the clinicopathological features of adrenocortical oncocytic tumors (ACOT) and to compare the diagnostic values of Lin-Weiss-Bisceglia (LWB) score and Helsinki...
To study the clinicopathological features of adrenocortical oncocytic tumors (ACOT) and to compare the diagnostic values of Lin-Weiss-Bisceglia (LWB) score and Helsinki score. Forty-four cases of ACOT diagnosed at Beijing Friendship Hospital, China from March 2008 to July 2019 were histologically analyzed to evaluate their malignant potential (benign versus malignant) according to two scoring criteria. Immunohistochemical studies (EnVision method) were also used. There were 23 males and 21 females with an average age of 46 years. Histologically, the tumor cells were arranged in trabecular, chrysanthemum-shaped, glandular and microcapsule structures, while clear cells were rare or absent. Most of the tumor cells were moderately atypical, and intranuclear inclusion bodies were conspicuous. Immunohistochemical staining showed that tumor cells were positive for Melan A, inhibin, Syn and calretinin. The average proliferation index was 3% in benign ACOT, about 5% in ACOT of malignant potential, and>20% in malignant ACOT. According to the LWB score, 61.4% (27/44) of the tumors were on the left side and had multiple lesions. The percentage of benign ACOT was 59.1% (26/44), malignant potential 6.8% (3/44), malignant 34.1% (15/44), respectively. Among the 15 malignant ACOT, the mitotic figures>5/50 HPF were found in 13 cases, necrosis in 11 cases and capsule invasion in 10 cases. According to the Helsinki score, 65.9% (29/44) of the tumors were benign, and 34.1% (15/44) were malignant. There was no significant difference between the two scoring standards (0.05). During the follow-up of 9 to 144 months, 31 patients survived without disease and 13 patients relapsed or had metastasis. ACOT more likely be benign than malignant. The left side is more common. Malignant tumors are prone to recurrence and metastasis. The morphological parameters (high mitotic index, necrosis, and capsular invasion) in the LWB scoring standards combined with immunohistochemical parameters (Ki-67) in the Helsinki score are helpful for the diagnosis of malignant ACOT and are important predictors of poor prognosis.
Topics: Adrenal Cortex Neoplasms; Biomarkers, Tumor; China; Female; Humans; Male; Middle Aged; Mitotic Index; Necrosis; Neoplasms, Glandular and Epithelial
PubMed: 33915658
DOI: 10.3760/cma.j.cn112151-20210209-00142 -
Anticancer Research Jan 2021We aimed to develop a rapid, simple procedure and an algorithm for quantitative analysis and classification of the metastatic risk of gastrointestinal stromal tumours...
AIM
We aimed to develop a rapid, simple procedure and an algorithm for quantitative analysis and classification of the metastatic risk of gastrointestinal stromal tumours (GIST) for clinical use.
MATERIALS AND METHODS
Eighteen specimens from laparoscopic local gastrectomy were assessed by flow cytometry. We devised a new risk classification for GIST by combining flow cytometry parameters with tumour size and evaluated whether the combined parameters correlated with the modified Fletcher risk classification.
RESULTS
We found a significant correlation between clinical prognostic factors (mitotic count and Ki-67 labelling index) and the flow cytometry parameters DNA ploidy, DNA index and S-phase fraction. The combined parameters established from tumour size and the flow cytometry parameters showed a high correlation with the modified Fletcher risk classification (p=0.0064). Flow cytometry had to be performed for approximately 10 minutes to determine the metastatic risk.
CONCLUSION
Rapid flow cytometry parameters can classify risk without the need for histological analysis.
Topics: Aged; Biomarkers; DNA, Neoplasm; Female; Flow Cytometry; Gastrointestinal Stromal Tumors; Humans; Ki-67 Antigen; Male; Middle Aged; Mitotic Index; Ploidies; Prognosis; Reproducibility of Results; Tumor Burden
PubMed: 33419806
DOI: 10.21873/anticanres.14758 -
Ecotoxicology and Environmental Safety Dec 2015Rare earth elements such as lanthanum (La) have been used as agricultural inputs in some countries in order to enhance yield and improve crop quality. However, little is...
Rare earth elements such as lanthanum (La) have been used as agricultural inputs in some countries in order to enhance yield and improve crop quality. However, little is known about the effect of La on the growth and structure of soybean, which is an important food and feed crop worldwide. In this study, bioaccumulation of La and its effects on the growth and mitotic index of soybean was evaluated. Soybean plants were exposed to increasing concentrations of La (0, 5, 10, 20, 40, 80, and 160 µM) in nutrient solution for 28 days. Plant response to La was evaluated in terms of plant growth, nutritional characteristics, photosynthetic rate, chlorophyll content, mitotic index, modifications in the ultrastructure of roots and leaves, and La mapping in root and shoot tissues. The results showed that the roots of soybean plants can accumulate sixty-fold more La than shoots. La deposition occurred mainly in cell walls and in crystals dispersed in the root cortex and in the mesophyll. When La was applied, it resulted in increased contents of some essential nutrients (i.e., Ca, P, K, and Mn), while Cu and Fe levels decreased. Moreover, low La concentrations stimulated the photosynthetic rate and total chlorophyll content and lead to a higher incidence of binucleate cells, resulting in a slight increase in roots and shoot biomass. At higher La levels, soybean growth was reduced. This was caused by ultrastructural modifications in the cell wall, thylakoids and chloroplasts, and the appearance of c-metaphases.
Topics: Cell Wall; Chlorophyll; Chloroplasts; Lanthanum; Mitotic Index; Photosynthesis; Plant Leaves; Plant Roots; Plant Shoots; Glycine max
PubMed: 26232040
DOI: 10.1016/j.ecoenv.2015.07.020 -
Archives of Biochemistry and Biophysics Dec 2020β-Sitosterol (βSito) is the most abundant phytosterol found in vegetable oils, grains such as wheat, beans, and corn, and in many phytosterol-enriched foods. It is...
β-Sitosterol (βSito) is the most abundant phytosterol found in vegetable oils, grains such as wheat, beans, and corn, and in many phytosterol-enriched foods. It is prone to oxidation by reactive oxygen species, such as ozone, leading to the formation of oxyphytosterols. A better understanding regarding the biological effects and mechanism of action of oxyphytosterols is required since the beneficial and adverse side effects of these compounds on human health remain highly controversial. In this work, we investigated the biological effects of β-Secosterol (βSec), a new oxyphytosterol generated by the reaction of βSito with ozone. Treatment of HepG2 cells with βSito or βSec (0.1-100 μM) for 24, 48, and 72 h induced a dose-dependent reduction of cell viability in the MTT assay, with βSec showing higher efficacy than βSito. However, βSec presented a lower potency than βSito, showing IC = 37.32 μM, higher than βSito (IC = 0.23 μM) at 48 h. Cell cycle analyses by flow cytometry showed a slight decrease of G0/G1 phase with βSito 0.5 μM, but a significant cell cycle arrest at the G0/G1 phase in the treatment for 48 h with βSec 20 μM (62.69 ± 2.15%, p < 0.05) and βSec 40 μM (66.96 ± 5.39%, p < 0.0001) when compared to control (56.97 ± 2.60%). No suggestion of apoptosis was indicated by flow cytometry data. Also, βSec (20 and 40 μM) reduced the mitotic index. In the laser scanning confocal microscopy analysis no alterations in cell morphology were observed with βSito (0.5 μM). Nevertheless, round-shaped cells, abnormal nuclear morphology with shrinkage, and formation of microtubules clusters were observed in the treatment with βSec, indicating a disruption in the microtubules network organization. N-acetyl-l-cysteine was not able to inhibit any of these cellular effects, indicating a lack of involvement of oxidative stress in the mechanism of action of βSec. Although not further investigated in this study, it was discussed the hypothesis that covalent adduct formation with lysine residues of proteins, could play an important role in the biological effects elicited by βSec. Elucidation of the primary cellular processes induced by βSec provides the essential knowledge to be aware of its potential adverse side effects or therapeutic use of this oxyphytosterol.
Topics: Acetylcysteine; Cell Nucleus; Cell Survival; G1 Phase Cell Cycle Checkpoints; Hep G2 Cells; Humans; Microtubules; Mitotic Index; Oxidative Stress; Ozone; Sitosterols
PubMed: 33130087
DOI: 10.1016/j.abb.2020.108654 -
Environmental Science and Pollution... Jun 2020Imazalil (IMZ), a fungicide containing imidazole group, is extensively used for the prevention and treatment of fungal diseases in plants. Current study was performed to...
Imazalil (IMZ), a fungicide containing imidazole group, is extensively used for the prevention and treatment of fungal diseases in plants. Current study was performed to examine cyto-genotoxic potential of IMZ on Allium cepa roots by following Allium ana-telophase and single cell gel electrophoresis (comet) assays. The concentration which reduced the growth of the root tips of IMZ by 50% compared to the negative control group (EC) was found to be 1 μg/mL by Allium root growth inhibition test. 0.5, 1, and 2 μg/mL concentrations of IMZ were exposed to Allium roots for intervals of 24, 48, 72, and 96 h. 10 μg/mL of methyl methane sulfonate (MMS) and distilled water were used as control groups, both positive and negative. Statistical analysis was performed by using one-way ANOVA with Duncan's multiple comparison tests at p ≤ 0.05 and Pearson correlation test at p = 0.01. IMZ showed cytotoxic effect by statistically decreasing root growth and mitotic index (MI) and also genotoxic effect by statistically increasing chromosomal aberrations (CAs) and DNA damage compared to the negative control group. With these cyto-genotoxic effects, it should be used carefully and further cyto-genotoxic mechanisms should be investigated along with other toxicity tests.
Topics: Chromosome Aberrations; Cytogenetic Analysis; DNA Damage; Fungicides, Industrial; Humans; Imidazoles; Meristem; Mitotic Index; Onions; Plant Roots
PubMed: 32242316
DOI: 10.1007/s11356-020-08553-2 -
Scientific Reports Sep 2022Trifloxystrobin (TFS) is a strobilurin-type fungicide that should be investigated due to its risks to non-targeted organisms. The goal of this study was to assess the...
Trifloxystrobin (TFS) is a strobilurin-type fungicide that should be investigated due to its risks to non-targeted organisms. The goal of this study was to assess the susceptibility of Allium cepa L. to TFS in a multi-pronged approach. For 72 h, 0.2 g/L, 0.4 g/L and 0.8 g/L doses of TFS were administered to A. cepa bulbs and the control group was treated with tap water. The toxic effects of TFS were tested, considering physiological, cytogenetic, biochemical and anatomical analyses. TFS delayed growth by reducing the rooting ratio, root elongation and weight increase. Following TFS treatments, mitotic index (MI) scores decreased, while the formation of micronucleus (MN) and chromosomal aberrations (CAs) ascended. CAs types induced by TFS were listed according to their frequency as fragment, vagrant chromosome, sticky chromosome, uneven distribution of chromatin, bridge, nucleus with vacuoles, reverse polarization and irregular mitosis. TFS provoked an increment in superoxide dismutase (SOD) and catalase (CAT) enzyme activities as well as an accumulation of malondialdehyde (MDA). Meristematic cells of A. cepa roots treated with TFS had various anatomical damages, including damaged epidermis, flattened cell nucleus, damaged cortex and thickness in the cortex cell wall. All damages arising from TFS treatments exhibited dose-dependency. The findings of the present study revealed the serious toxicity of TFS in a non-targeted plant. It should not be neglected to evaluate the potential hazards of TFS with different toxicity tests.
Topics: Acetates; Allium; Antioxidants; Chromosome Aberrations; Fungicides, Industrial; Imines; Malondialdehyde; Meristem; Mitotic Index; Onions; Plant Roots; Strobilurins
PubMed: 36076029
DOI: 10.1038/s41598-022-19571-0 -
Modern Pathology : An Official Journal... Jul 2021Ki67, a nuclear proliferation-related protein, is heavily used in anatomic pathology but has not become a companion diagnostic or a standard-of-care biomarker due to...
Ki67, a nuclear proliferation-related protein, is heavily used in anatomic pathology but has not become a companion diagnostic or a standard-of-care biomarker due to analytic variability in both assay protocols and interpretation. The International Ki67 Working Group in breast cancer has published and has ongoing efforts in the standardization of the interpretation of Ki67, but they have not yet assessed technical issues of assay production representing multiple sources of variation, including antibody clones, antibody formats, staining platforms, and operators. The goal of this work is to address these issues with a new standardization tool. We have developed a cell line microarray system in which mixes of human Karpas 299 or Jurkat cells (Ki67) with Sf9 (Spodoptera frugiperda) (Ki67) cells are present in incremental standardized ratios. To validate the tool, six different antibodies, including both ready-to-use and concentrate formats from six vendors, were used to measure Ki67 proliferation indices using IHC protocols for manual (bench-top) and automated platforms. The assays were performed by three different laboratories at Yale and analyzed using two image analysis software packages, including QuPath and Visiopharm. Results showed statistically significant differences in Ki67 reactivity between each antibody clone. However, subsets of Ki67 assays using three clones performed in three different labs show no significant differences. This work shows the need for analytic standardization of the Ki67 assay and provides a new tool to do so. We show here how a cell line standardization system can be used to normalize the staining variability in proliferation indices between different antibody clones in a triple negative breast cancer cohort. We believe that this cell line standardization array has the potential to improve reproducibility among Ki67 assays and laboratories, which is critical for establishing Ki67 as a standard-of-care assay.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cell Line, Tumor; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Middle Aged; Mitotic Index; Triple Negative Breast Neoplasms
PubMed: 33536573
DOI: 10.1038/s41379-021-00745-6 -
Drug and Chemical Toxicology Jul 2019Paraben is a phenolic derivative of benzoic acid extensively used as preservatives in food, pharmaceutical, and cosmetic industries due to its antimicrobial...
Paraben is a phenolic derivative of benzoic acid extensively used as preservatives in food, pharmaceutical, and cosmetic industries due to its antimicrobial characteristics. The objective of this study was to evaluate the in vitro genotoxic effects of paraben in human lymphocyte cultures. Cells were analyzed by cytokinesis-block micronucleus (CBMN), chromosome aberration (CA), sister chromatid exchange (SCE), and comet tests. For CBMN, CA, and SCE assays, the human lymphocytes were isolated from healthy donors and incubated with 500, 250, 100, and 50 µg/mL of paraben for 24 and 48 h, and for comet assay, cells were exposed to 1000, 750, 500, and 250 µg/mL of paraben for an hour. Results showed that numbers of MN and SCEs were not significant in the cells exposed to paraben when compared to the solvent control. However, 500 and 250 µg/mL of paraben induced the CA after 24 h. Also, we observed a significant decrease in the cytokinesis-block proliferation index in cells exposed 250-500 µg/mL paraben for 24 h, and 100, 250, and 500 µg/mL for 48 h. The mitotic index was also decreased at all concentrations and periods. However, the proliferation index was statistically decreased at all concentrations after 48 h treatments. Only the highest concentration of paraben caused DNA migration (mean tail length) in human lymphocytes analyzed by Comet assay. Taken together, results indicated that paraben had cytotoxic effects and caused genotoxicity by affecting directly chromosomes and DNA in human lymphocyte cells in vitro, and may have genotoxic potential for human.
Topics: Cells, Cultured; Chromosome Aberrations; Comet Assay; DNA Damage; Dose-Response Relationship, Drug; Healthy Volunteers; Humans; Lymphocytes; Micronuclei, Chromosome-Defective; Micronucleus Tests; Mitotic Index; Mutagens; Parabens; Sister Chromatid Exchange
PubMed: 29281926
DOI: 10.1080/01480545.2017.1414834 -
Journal of Environmental Science and... 2022Studies assessing the toxicity of glyphosate and 2,4-dichlorophenoxyacetic acid mixture are scarce. The aim of this study was to evaluate the cytotoxicity and...
Studies assessing the toxicity of glyphosate and 2,4-dichlorophenoxyacetic acid mixture are scarce. The aim of this study was to evaluate the cytotoxicity and genotoxicity of the mixture of these herbicides using . Roots were exposed to glyphosate (1.56 and 11.66 mg mL), 2,4-D (0.28 and 17.5 mg mL) and mixture for 24 h, based on the average concentration applied in the field and the acute reference dose (ARfD) established in Brazil. Both isolated and associated herbicides induced a significative decrease in mitotic index (MI) ( < 0.0001) in all tested concentrations. Regarding the genotoxicity results, 2,4-D and the mixture showed, at concentrations applied in the field, a significative increase of chromosomal anomalies (CA) index compared to control ( < 0.0001) and glyphosate ( = 0.024 and = 0.0002, respectively). All tested groups from the ARfD showed a significative difference compared to the control group ( < 0.0001), as well as glyphosate and 2,4-D isolated compared to the mixture ( = 0.0005 and < 0.0001, respectively). The most observed CA were apoptotic bodies, giant cells, and nuclear erosions. We emphasize the need for further studies assessing the toxicity of these herbicides' mixture due to the distinct effects caused in different organisms.
Topics: 2,4-Dichlorophenoxyacetic Acid; Biological Assay; Chromosome Aberrations; DNA Damage; Glycine; Herbicides; Mitotic Index; Onions; Plant Roots; Glyphosate
PubMed: 35112655
DOI: 10.1080/03601234.2022.2034432