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Ecotoxicology and Environmental Safety Jan 2020In the last few decades, tremendous increase in the use of wireless electronic gadgets, particularly the cell phones, has significantly enhanced the levels of...
In the last few decades, tremendous increase in the use of wireless electronic gadgets, particularly the cell phones, has significantly enhanced the levels of electromagnetic field radiations (EMF-r) in the environment. Therefore, it is pertinent to study the effect of these radiations on biological systems including plants. We investigated comparative cytotoxic and DNA damaging effects of 900 and 1800 MHz EMF-r in Allium cepa (onion) root meristematic cells in terms of mitotic index (MI), chromosomal aberrations (CAs) and single cell gel electrophoresis (comet assay). Onion bulbs were subjected to 900 and 1800 MHz (at power densities 261 ± 8.50 mW m and 332 ± 10.36 mW m, respectively) of EMF-r for 0.5 h, 1 h, 2 h, and 4 h. Root length declined by 13.2% and 12.3%, whereas root thickness was increased by 46.7% and 48.3% after 4 h exposure to 900 MHz and 1800 MHz, respectively. Cytogenetic studies exhibited clastogenic effect of EMF-r as depicted by increased CAs and MI. MI increased by 36% and 53% after 2 and 4 h exposure to 900 MHz EMF-r, whereas it increased by 41% and 67% in response to 1800 MHz EMF-r. Aberration index was increased by 41%-266% and 14%-257% during 0.5-4 h of exposure to 900 MHz and 1800 MHz, respectively, over the control. EMF-r exposure decreased % head DNA (DNAH) and increased % tail DNA (DNAT) and olive tail moment (OTM) at both 900 and 1800 EMF-r. In 4 h exposure treatments, head DNA (%) declined by 19% and 23% at 900 MHz and 1800 MHz, respectively. DNAT and OTM were increased by 2.3 and 3.7 fold upon exposure to 900 MHz EMF-r over that in the control, whereas 2.8 and 5.8 fold increase was observed in response to 1800 MHz EMF-r exposure for 4 h and the difference was statistically significant. The study concludes that EMF-r in the communication range (900 and 1800 MHz) adversely affect root meristems in plants and induce cytotoxic and DNA damage. EMF-r induced DNA damage was more pronounced at 1800 MHz than that at 900 MHz.
Topics: Cell Phone; Chromosome Aberrations; Comet Assay; DNA Damage; Dose-Response Relationship, Radiation; Electromagnetic Fields; Electromagnetic Radiation; Meristem; Mitotic Index; Onions; Time Factors
PubMed: 31698176
DOI: 10.1016/j.ecoenv.2019.109786 -
Environmental Science and Pollution... Dec 2022Phytotoxicity, cytotoxicity and genotoxicity of pesticides with various mechanisms of targeted activity were studied in a hydroponic culture of 2-day-old seedlings of...
Phytotoxicity, cytotoxicity and genotoxicity of pesticides with various mechanisms of targeted activity were studied in a hydroponic culture of 2-day-old seedlings of Triticum aestivum. All studied pesticides (with the exception of metribuzin) exhibited dose-dependent phytotoxicity (inhibited the growth of the main root and reduced the yield of root biomass). All studied pesticides did not affect mitotic index in the root apex meristem but did affect the duration of some phases of mitosis. Herbicides increased, while fungicides, on the contrary, decreased the duration of the cytokinesis phase. All pesticides (1 μg/mL) exhibited genotoxic activity: in the root apex meristem the number of cells with mitotic abnormalities was significantly higher than in the control variant (7-14 times). The genotoxic activity of metribuzin and tebuconazole was 2 times lower than for tribenuron-methyl, fenoxaprop-P-ethyl, epoxiconazole and azoxystrobin. The genotoxicity of the studied pesticides was combined: depending on the class of the pesticide, clastogenic or aneugenous effects dominated.
Topics: Meristem; Triticum; Herbicides; Fungicides, Industrial; Pesticides; Chromosome Aberrations; Mitotic Index; Plant Roots
PubMed: 35818017
DOI: 10.1007/s11356-022-21936-x -
Virchows Archiv : An International... Jan 2019Mitotic figure (MF) counting is important in the evaluation of meningioma grading. Nevertheless, mitosis assessment on hematoxylin and eosin (H&E)-stained slides may be... (Comparative Study)
Comparative Study
Mitotic figure (MF) counting is important in the evaluation of meningioma grading. Nevertheless, mitosis assessment on hematoxylin and eosin (H&E)-stained slides may be problematic because of technical factors and pathologist's experience. Phosphohistone H3 (PHH3) is a mitosis-specific antibody that has proven to facilitate mitotic count in various tumors. However, the antibody performance between PHH3 serine10 (S10) and serine28 (S28) has never been compared in these tumors before. In this study, 48 cases of meningioma (28 grade I, 14 grade II, 6 grade III) were evaluated using immunohistochemical stains for four commercially available PHH3 (S10) and S28 antibodies to identify MFs and validate PHH3 intra- and interobserver reproducibility and agreement. Two pathologists counted MFs on both H&E- and PHH3-stained slides. H&E and PHH3 MFs were highly correlated (Spearman's rho = 0.96 for PHH3 (S10)-Biocare, 0.96 for PHH3 (S10)-CST, 0.91 for PHH3 (S28)-Abcam, and 0.89 for PHH3 (S28)-Santa Cruz. The mean difference between an H&E and PHH3 mitotic count is 0.81 for PHH3 (S10)-Biocare, 0.95 for PHH3 (S10)-CST, - 0.97 for PHH3 (S28)-Abcam, and - 0.97 for PHH3 (S28)-Santa Cruz. For comparison among four PHH3 antibodies, PHH3 mitotic counts had both a good intra- and interobserver reproducibility (p > 0.05). Regarding to World Health Organization (WHO) grade, there was not a significant discrepancy in the stratification of tumor grades for all four PHH3 antibodies in terms of interobserver agreement. The Cohen's kappa coefficient (K) was 0.93 for PHH3 (S10)-Biocare, 0.82 for PHH3 (S10)-CST, 0.76 for PHH3 (S28)-Abcam, and 0.80 for PHH3 (S28)-Santa Cruz. Considering survival analyses, all five proliferation indices were univariately associated with recurrences. Increased PHH3 mitotic indices (MIs) were significantly associated with recurrence-free survival in univariate Cox proportional hazards regression analysis (p < 0.001) and remained an independent predictor in multivariate analysis (p < 0.05). The appropriate prognostic cutoff values for recurrence prediction were 5 or more per 10 high-power fields (HPFs) for PHH3 (S10) and 3 or more per 10 HPFs for PHH3 (S28).
Topics: Antibodies; Antibody Specificity; Biomarkers, Tumor; Histones; Humans; Immunohistochemistry; Meningeal Neoplasms; Meningioma; Mitosis; Mitotic Index; Neoplasm Grading; Observer Variation; Phosphorylation; Predictive Value of Tests; Progression-Free Survival; Reproducibility of Results; Risk Factors; Time Factors; Treatment Outcome
PubMed: 30267302
DOI: 10.1007/s00428-018-2458-2 -
Histopathology Feb 2023Mitotic count in breast cancer is an important prognostic marker. Unfortunately, substantial inter- and intraobserver variation exists when pathologists manually count...
Improving mitotic cell counting accuracy and efficiency using phosphohistone-H3 (PHH3) antibody counterstained with haematoxylin and eosin as part of breast cancer grading.
BACKGROUND
Mitotic count in breast cancer is an important prognostic marker. Unfortunately, substantial inter- and intraobserver variation exists when pathologists manually count mitotic figures. To alleviate this problem, we developed a new technique incorporating both haematoxylin and eosin (H&E) and phosphorylated histone H3 (PHH3), a marker highly specific to mitotic figures, and compared it to visual scoring of mitotic figures using H&E only.
METHODS
Two full-face sections from 97 cases were cut, one stained with H&E only, and the other was stained with PHH3 and counterstained with H&E (PHH3-H&E). Counting mitoses using PHH3-H&E was compared to traditional mitoses scoring using H&E in terms of reproducibility, scoring time, and the ability to detect mitosis hotspots. We assessed the agreement between manual and image analysis-assisted scoring of mitotic figures using H&E and PHH3-H&E-stained cells. The diagnostic performance of PHH3 in detecting mitotic figures in terms of sensitivity and specificity was measured. Finally, PHH3 replaced the mitosis score in a multivariate analysis to assess its significance.
RESULTS
Pathologists detected significantly higher mitotic figures using the PHH3-H&E (median ± SD, 20 ± 33) compared with H&E alone (median ± SD, 16 ± 25), P < 0.001. The concordance between pathologists in identifying mitotic figures was highest when using the dual PHH3-H&E technique; in addition, it highlighted mitotic figures at low power, allowing better agreement on choosing the hotspot area (k = 0.842) in comparison with standard H&E (k = 0.625). A better agreement between image analysis-assisted software and the human eye was observed for PHH3-stained mitotic figures. When the mitosis score was replaced with PHH3 in a Cox regression model with other grade components, PHH3 was an independent predictor of survival (hazard ratio [HR] 5.66, 95% confidence interval [CI] 1.92-16.69; P = 0.002), and even showed a more significant association with breast cancer-specific survival (BCSS) than mitosis (HR 3.63, 95% CI 1.49-8.86; P = 0.005) and Ki67 (P = 0.27).
CONCLUSION
Using PHH3-H&E-stained slides can reliably be used in routine scoring of mitotic figures and integrating both techniques will compensate for each other's limitations and improve diagnostic accuracy, quality, and precision.
Topics: Humans; Female; Eosine Yellowish-(YS); Mitotic Index; Breast Neoplasms; Hematoxylin; Reproducibility of Results; Biomarkers, Tumor; Immunohistochemistry; Mitosis; Antibodies; Phosphorylation
PubMed: 36349500
DOI: 10.1111/his.14837 -
PloS One 2021In this research, eight local mung bean (Vigna radiata) varieties were analyzed for their performance against two levels of CdCl2 solution (0.3 and 0.5 mM) alone and...
In this research, eight local mung bean (Vigna radiata) varieties were analyzed for their performance against two levels of CdCl2 solution (0.3 and 0.5 mM) alone and priming with gibberellic acid (GA3) (100 μM), salicylic acid (SA) (50 μM) and proline (5 mM) solution prior to Cd exposure. Mung bean seedlings were analyzed for disturbance in cytological, morphological, biochemical and enzymatic parameters under cadmium stress. For cytological studies, 48 h grown mung bean seedlings root tips were used to prepare slides and studied for percent mitotic index (MI%) and to calculate percent C-mitosis, laggard, sticky and fragmented chromosomes, pictures were captured by a Nikon camera (DS-Fi 1 Japan) attached with a microscope. One-week grown mung seedlings were studied for growth traits, malondialdehyde (MDA), protein, proline and antioxidant enzymes. ANOVA and DMR test of this research revealed that all the tested mung bean varieties and treatments were significantly different regarding mitotic index and number of chromosomal aberrations. Both the Cd treatments exhibited increased total chromosomal aberrations with different types and a maximum decrease in MI%. In pretreated samples, GA3, SA and proline serve as mitigating agents that reduce mutagenic effects of Cd in mung bean by increasing MI% and decreasing chromosomal aberrations as compared to non-pretreated samples. Both the Cd treatments showed a decrease in all growth traits. Total proteins were also found to be significantly reduced in a dose-dependent manner in all genotypes. Cd treatment increased the activities of all antioxidant enzymes tested. Cd caused oxidative damage as indicated by elevated levels of MDA content in treated samples in comparison to control. Proline content levels were also high in Cd treated seedlings indicating stress. Results demonstrated that pretreatment with phytohormones and proline before Cd were found to improve all morphological parameters, by altering antioxidant enzymes activities along with a decrease in MDA and proline contents as well. It was further noticed that the performance of GA3 was better at 0.3 mM Cd treatment while SA was found to be a good mitigating agent at 0.5 mM Cd stress in all tested mung bean varieties. This research concluded less deleterious effects of Cd on AZRI-2006 while more sensitivity to NM-51 towards Cd. Priming with phytohormones and proline is a user-friendly, economical, and simple mitigation strategy to reduce Cd toxicity in plants and get better yield from contaminated lands.
Topics: Acclimatization; Cadmium Chloride; DNA, Plant; Malondialdehyde; Mitotic Index; Oxidative Stress; Plant Growth Regulators; Plant Roots; Proline; Salicylic Acid; Seedlings; Vigna
PubMed: 34587203
DOI: 10.1371/journal.pone.0257924 -
Protoplasma Jul 2023Diterpenoid 3-epicaryoptin (CHO) is abundant in the leaves of Clerodendrum inerme, a traditionally used medicinal plant, and has insect antifeedant activities. Here, we...
Diterpenoid 3-epicaryoptin (CHO) is abundant in the leaves of Clerodendrum inerme, a traditionally used medicinal plant, and has insect antifeedant activities. Here, we aim to explore the cytogenotoxic effects of compound 3-epicaryoptin in Allium cepa root apical meristem cells. 3-epicaryoptin (concentrations of 100, 150, and 200 µg mL) and the standard compound colchicine (200 µg mL) were applied to A. cepa roots for 2, 4, and 4 + 16 h (4-h treatment followed by 16-h recovery). Cytogenotoxicity was analyzed by studying the root growth retardation (RGR), mitotic index (MI), and chromosomal aberrations. The result showed statistically significant (p < 0.01), concentration-dependent RGR effects of 3-epicaryoptin treatment compared with the negative control. A study of cell frequency in different phases of cell division observed a significant (p < 0.001) increase in the metaphase cell percentage (66.2 ± 0.58%, 150 µg mL), which subsequently caused an increase in the frequency of MI (12.29 ± 0.34%, 150 µg mL) at 4 h of 3-epicaryoptin treatment and that was comparable with the colchicine action. The cytological study revealed that the 3-epicaryoptin treatment could induce different types of chromosomal abnormalities, such as colchicine-like metaphase, vagrant chromosomes, sticky chromosomes, anaphase bridge, lagging chromosomes, multipolar anaphase-telophase, and an increased frequency of micronuclei and polyploid cells. These findings indicate that 3-epicaryoptin is cytogenotoxic, and thus, C. inerme should be used with caution in traditional medicine.
Topics: Meristem; Onions; Plant Roots; Mitosis; Mitotic Index; Chromosome Aberrations; DNA Damage
PubMed: 36735079
DOI: 10.1007/s00709-023-01838-6 -
Modern Pathology : An Official Journal... Sep 2020Pulmonary neuroendocrine neoplasms are classified by WHO as either typical or atypical carcinoids, large cell (LCNEC) or small cell (SCLC) neuroendocrine carcinoma based...
Pulmonary neuroendocrine neoplasms are classified by WHO as either typical or atypical carcinoids, large cell (LCNEC) or small cell (SCLC) neuroendocrine carcinoma based on mitotic count, morphology, and necrosis assessment. LCNEC with low mitotic count and sharing morphologic features with carcinoids are in a gray zone for classification and their rare prevalence and the paucity of studies precludes proper validation of the current grading system. In this study, we aim to investigate their clinicopathological and transcriptomic profiles. Lung resection specimens obtained from 18 patients diagnosed with carcinoids or LCNEC were selected. Four of them were characterized as borderline tumors based on a mitotic rate ranging between 10 and 30 mitoses per 2 mm. Comprehensive morphological and immunohistochemical (IHC) evaluation was performed and tumor-based transcriptomic profiles were analyzed through unsupervised clustering. Clustering analysis revealed two distinct molecular groups characterized by low (C1) and high (C2) proliferation. C1 was comprised of seven carcinoids and three borderline tumors, while C2 was comprised of seven LCNEC and one borderline tumor. Furthermore, patients in cluster C1 had a better recurrence-free survival compared with patients in cluster C2 (20% vs 75%). Histological features, IHC profile, and molecular analysis showed that three out of four borderline tumors showed features consistent with carcinoids. Therefore, our findings convey that the current diagnostic guidelines are suboptimal for classification of pulmonary neuroendocrine tumors with increased proliferative index and carcinoid-like morphology. These results support the emerging concept that neuroendocrine tumors with carcinoid-like features and mitotic count of <20 mitoses per 2 mm should be regarded as pulmonary carcinoids instead of LCNEC.
Topics: Aged; Biomarkers, Tumor; Carcinoid Tumor; Female; Gene Expression Regulation, Neoplastic; Humans; Lung; Lung Neoplasms; Male; Middle Aged; Mitosis; Mitotic Index; Retrospective Studies; Transcriptome
PubMed: 32291397
DOI: 10.1038/s41379-020-0538-8 -
The American Surgeon Nov 2016With the increasing popularity of sleeve gastrectomy, many stomach specimens are being evaluated. Understanding the significance and treatment for unexpected pathology...
With the increasing popularity of sleeve gastrectomy, many stomach specimens are being evaluated. Understanding the significance and treatment for unexpected pathology is important. This study examines the incidence of relevant histopathology of sleeve gastrectomy specimens. It evaluates previous data for each histopathology and provides recommendations for treatment. In this study, a retrospective review was performed for 241 patients who underwent sleeve gastrectomy from 2009 to 2014 at a single institution. Of the specimens, 122 had no significant histopathology, 91 had gastritis, 13 had lymphoid aggregates, 5 had hyperplasia, 3 had intestinal metaplasia, 3 had gastrointestinal stromal tumors (GISTs), and 3 had gastric polyps. Of the GISTs all had a low mitotic rate and the size of the tumor ranged from 1.5 to 4.5 cm. The findings of metaplasia may be a marker for increased risk of malignancy and may require additional surveillance. The findings of GIST may warrant interval imaging to survey for recurrence, though the likelihood of recurrence for the tumors in this study is less than 2 per cent based on previous studies.
Topics: Adenomatous Polyps; Adult; Aged; Female; Gastrectomy; Gastritis; Gastrointestinal Stromal Tumors; Humans; Hyperplasia; Lymphoid Tissue; Male; Metaplasia; Middle Aged; Mitotic Index; Retrospective Studies; Stomach Neoplasms; Tumor Burden; Young Adult
PubMed: 28206938
DOI: No ID Found -
Histopathology Apr 2020Proliferation assessment by the use of Ki67 is a crucial component in intrinsic subtyping of luminal breast cancers (BCs), but suffers from variability between...
Luminal A versus luminal B breast cancer: MammaTyper mRNA versus immunohistochemical subtyping with an emphasis on standardised Ki67 labelling-based or mitotic activity index-based proliferation assessment.
AIMS
Proliferation assessment by the use of Ki67 is a crucial component in intrinsic subtyping of luminal breast cancers (BCs), but suffers from variability between laboratories, observers, and methods. MammaTyper is a quantitative molecular tool that measures mRNA levels of ERBB2, ESR1, PGR and MKI67 in BC, and interprets the results according to the St Gallen 2013 consensus recommendations. We compared MammaTyper with immunohistochemistry (IHC)-based subtypes, with a focus on standardised proliferation assessment.
METHODS AND RESULTS
We analysed the agreement in assigning subtypes between MammaTyper and receptor IHC in 101 unifocal luminal HER2-negative early BCs of no special type. Two Ki67 counting protocols, Ki67-Global (Ki67-G) and Ki67-HotSpot (Ki67-H), recommended by the International Ki67 in BC Working Group, and the mitotic activity index (MAI) were used for proliferation assessment. The proportions of BCs identified as luminal A and as luminal B were 55% and 45% for MammaTyper, 55% and 45% for IHC + Ki67-G, 36% and 64% for IHC + Ki67-H, and 56% and 44% for IHC + MAI. The levels of agreement between MammaTyper-based and IHC-based subtyping were 84% (κ = 0.679) for IHC + Ki67-G, 72% (κ = 0.462) for IHC + Ki67-H, and 89% (κ = 0.779) for IHC + MAI.
CONCLUSIONS
High rates of agreement between mRNA-based and IHC-based intrinsic subtyping of luminal HER2-negative BC can be achieved. However, the agreement between IHC-based and MammaTyper-based luminal subtypes depends on the proliferation assessment method, and was highest when the MAI was used. Further comparative clinical studies are needed to determine which method is to be preferred, including analysis of cost-effectiveness.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Cell Proliferation; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Middle Aged; Mitotic Index; RNA, Messenger
PubMed: 31846096
DOI: 10.1111/his.14048 -
Journal of Medical Case Reports Jan 2016Atypical uterine leiomyomas show benign behavior. However, the distinction between leiomyomas and leiomyosarcomas may at times be problematic. We report a rare case of... (Review)
Review
BACKGROUND
Atypical uterine leiomyomas show benign behavior. However, the distinction between leiomyomas and leiomyosarcomas may at times be problematic. We report a rare case of atypical uterine leiomyoma. We try to investigate potential immunohistochemical parameters that could be essential to distinguish cases of malignant smooth muscle tumors and those of uncertain or borderline histology.
CASE PRESENTATION
A 56-year-old white ethnic Albanian woman from Kosovo presented with uterine bleeding because of uterine multiple leiomyomas. A hysterectomy with unilateral adnexectomy was performed. Her hysterectomy specimen contained multiple leiomyomas in submucosal, intramural and subserosal locations. The leiomyomas were well demarcated, firm and white with a whorled cut surface and one had foci of hemorrhage. Histology of most of the leiomyomas showed a whorled (fascicular) pattern of smooth muscle bundles separated by well-vascularized connective tissue. Smooth muscle cells were elongated with eosinophilic or occasional fibrillar cytoplasm and distinct cell membranes. Some of them developed areas of degeneration including hyaline change, with less than five mitotic figures per ten high power fields in most mitotically active areas, and no significant atypia. One leiomyoma was characterized by moderately to severely pleomorphic atypical tumor cells with low mitotic counts and no coagulative tumor cell necrosis. Immunohistochemistry showed strong immunoreactivity for vimentin, smooth muscle actin and desmin, while cyclin-dependent kinase inhibitor 2A (p16), and B-cell lymphoma 2 (bcl-2) showed focal immunoreactivity, estrogen and progesterone were positive, Ki-67 expressed a low proliferation index, whereas p21 and tumor suppressor gene p53 were negative.
CONCLUSIONS
The combination of evaluation of conventional morphologic criteria with cyclin-dependent kinase inhibitor 2A (p16), p21, progesterone, B-cell lymphoma 2, tumor suppressor gene p53 and Ki-67 expression may be of great value in the assessment of uterine smooth muscle tumors of uncertain or borderline histology.
Topics: Diagnosis, Differential; Female; Humans; Leiomyomatosis; Leiomyosarcoma; Middle Aged; Mitotic Index; Uterine Neoplasms
PubMed: 26801982
DOI: 10.1186/s13256-016-0800-3