IN VITRO GENOTOXICITY OF THREE PLANTS FROM ASTERACEAE FAMILY IN HUMAN LYMPHOCYTES CULTURES

Background: Onopordum carduiforme, Centaurea verutum, and Achillea santolina are medicinal plants grown in Syria and commonly used in traditional medicine. Such as antibacterial, antioxidant and anticancer properties. However, the genotoxic effects of these plants have not been studied. Aim and objective: the aim of this study was to evaluate the genotoxic effects of hydroethanolic extracts of these plants on human lymphocyte cultures model by evaluating the cell proliferation, determination of mitotic index (MI), and their effects on chromosomes. Methods: the hydroethanolic extracts of the aerial parts of the three plants were extracted using an Ultrasonic bath. Then the genotoxic effects of hydroethanolic extracts of these plants on human lymphocyte cultures was conducting by determination of mitotic index (MI). Results: the results showed that all three plants decreased non-significantly the mean of mitotic index in comparison with negative control (normal MI) (p>0.05) at concentrations (1, 3, 5 mg/ml) and the mitotic index values ranged was between (2.25±0.07 and 3.3±0.28). However, C. verutum showed the lowest mitotic index (3±0.14 at 1 mg/ml) and (2.25±0.07 at 5 mg/ml), and did not induce chromatid or chromosome breaks or gaps. Conclusion: these preliminary results on cytotoxicity and mutagenicity of these plants provide valuable information about the safety of using them in alternative medicine. Peer Review History: Received: 11 November 2021; Revised: 13 December; Accepted: 28 December, Available online: 15 January 2022 Academic Editor: Dr. A.A. Mgbahurike, University of Port Harcourt, Nigeria, [email protected] UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Ahmad Najib, Universitas Muslim Indonesia, Makassar, Indonesia, [email protected] Dr. Dennis Amaechi, MrsFoluBabade Mini Estate , Flat 5 by Old Soldiers Quarter, Sabongari/Bwari, Abuja- Federal Capital Territory, Nigeria. [email protected] Dr. Sangeetha Arullappan, Universiti Tunku Abdul Rahman, Malaysia, [email protected] Similar Articles: A STUDY ON DIFFERENT PLANTS OF APOCYNACEAE FAMILY AND THEIR MEDICINAL USES STUDY LITERATION OF CHEMICAL CONTENTS OF SOME PLANTS THAT POTENTIALLY AS THE SOLAR SOWS EXPLORING THE ANTIPARASITIC ACTIVITY OF MEDICINAL PLANTS

Role of L-Ornithine in Mitigation of Salt Stress In Allium Cepa L.

Effects of L-ornithine (150 mg/l) on the germination, seedling growth, mitotic index, chromosome aberrations and micronucleus frequency of Allium cepa L. bulbs germinated at 0.125 M salinity were studied. The radicle number of the group III bulbs germinated in the medium with ornithine alone as compared to ones of the group I (control) bulbs which germinated in distilled water medium. But, their germination percentage, radicle length and fresh weight were statistically the same as ones of the group I bulbs. Besides, the micronucleus frequency and chromosomal abnormalities in the root-tip meristematic cells of the group III bulbs showed increased germination compared to ones of the group I bulbs. However, their mitotic index statistically showed the same value as the group I bulbs. Salt stress significantly inhibited the germination and seedling growth of A. cepa bulbs. Moreover, it reduced the mitotic index in the root-meristem cells of the bulbs and fairly increased the number of chromosome aberrations and micronucleus frequency. On the other hand, the inhibitive effect of salt on the germination, seedling growth, mitotic index and micronucleus frequency was dramatically alleviated in varying degrees by ornithine application. But, it was ineffective in reducing the detrimental effect of salinity on the chromosome aberrations. The germination percentage, radicle lenght, radicle number, fresh weight, mitotic index, micronucleus frequency and chromosomal aberrations of the group II seedlings grown in 0.125 M salinity were 27%, 13.5 mm, 18.4, 7.1 g, 5.5, 18.3 and 60.8%, respectively while these values became 68%, 16.4 mm, 16.4, 10.5 g, 15.6, 7.6 and 74.8% in the group IV seedlings treated with L-ornithine. Bangladesh J. Bot. 50(4): 1165-1171, 2021 (December)

Summary of four scientific studies on Arsenicum album high dilution effect against Arsenic intoxication in mice

Background: Groundwater arsenic affects millions of people in about 20 countries. In West Bengal (India) and Bangladesh alone over 100 million people are exposed. The arsenic concentration in contaminated groundwater in Bangladesh was above the maximum permissible level of 0.05 mg/l as recommended by WHO for developing countries [1]. Drinking water is not the only source of poisoning. In arsenic contaminated areas, crops, vegetables, cereals, poultry, cattle, etc, also contain traces of arsenic. Chronic arsenic intoxication has been associated with several diseases such as melanosis, leuco-melanosis, hyperkeratosis, oedema, skin cancer… Cazin et al [2], have demonstrated the effect of high dilutions of arsenic compounds. They noted increased arsenic elimination from blood through urine and faeces in intoxicated rats. According to these research, the aim of Khuda Buksh studies [3-4-5] was to investigate whether high dilution Arsenicum album have any effect on arsenic accumulation in different tissues and to understand also how this high dilution could produce a protective effect on all the different organs. Methodology: Firstly, the effect of Arsenicum album 30 cH on the amount of arsenic accumulation was determined by spectrophotometric analysis in four tissues namely liver, kidney and testis in mice intoxicated by arsenic. The protective effect in chronic and acute arsenic intoxicated mice of Arsenicum Album 6cH, 30cH and 200cH has been evaluated using not only the activities of enzymatic and biomarker toxicity (aspartate amino transferase (AST), alanine amino transferase (ALT), acid phosphatase (AcP), alkaline phosphatase (AlkP), lipid peroxidation (LPO) and reduced glutathione (GSH)) but also the cytogenetical parameters (chromosome aberrations (CA), mitotic index (MI), sperm head anomaly (SHA) etc., ). Because, it is well demonstrated that these enzymes biomarkers reflect the degree of hepatotoxicity and oxidative stress caused by arsenic intoxication. Results: Compared to controls, Arsenicum album 30cH induced a significant decrease in accumulation of arsenic in 4 tissues namely liver, spleen, kidney and testis in intoxicated mice. In addition, both Arsenicum album 6cH, 30cH and 200cH reduced chromosome aberrations, sperm head abnormality frequencies and activities of acid and alkaline phosphatases, aspartate and alanine aminotransferases and lipid peroxidation, while mitotic index and activities of glutathione, catalase and succinate dehydrogenase were increased compared to controls. Conclusion: Altogether, theses results provide evidence of protective potentials of the Arsenicum album dilution against acute and chronic arsenic intoxication in mice. They also offer a new hypothesis that the mechanism of the homeopathic dilution could act through regulation of expression of certain genes. This explanation seems to be plausible because all biomarker tests are regulated by specific genetic regulatory mechanisms [6]. keywords: Arsenicum album, arsenic intoxication, enzymatic and biomarker toxicity. References: [1] WHO. WHO Guidelines for Drinking Water Quality, Vol. 2, 2nd edition. Geneva: WHO, 1996, 940–9. [2] Cazin JC, Cazin M, Gaborit JL, Chaoui A, Boiron J, Belon P, et al. A study of the effect of decimal and centesimal dilutions of arsenic on the retention and mobilization of arsenic in the rat. Hum Toxicol 1987;6:315–20. [3] Mitra K, Kundu SN, Khuda-Bukhsh AR. Efficacy of a potentized homoeopathic drug (Arsenicum Album-30) in reducing toxic effects produced by of arsenic trioxide in mice. I. On rate accumulation of arsenic in certain vital organs. Comp Ther Med 1998;6:178–84. [4] Pathikrit Banerjee. Evidences of Protective Potentials of Microdoses of Ultra-high Diluted Arsenic Trioxide in Mice Receiving Repeated Injections of Arsenic Trioxide. eCAM 2009; 1-10. [5] Pathikrit Banerjee, Comparative Efficacy of Two Microdoses of a Potentized Homeopathic Drug, Arsenicum Album, to Ameliorate Toxicity Induced by Repeated Sublethal Injections of Arsenic Trioxide in Mice. Pathobiology 2008;75:156–170. [6]/ Khuda-Bukhsh AR. Potentized homeopathic drugs act through regulation of gene expression: a hypothesis to explain their mechanism and pathways of action in vivo. Comp Ther Med 1997;5:43–6

Predictive value of morphological features and proliferative markers (mitotic index, Ki-67, PHH3) in bronchopulmonary neuroendocrine tumors

The aim of this study was to evaluate relations of mitotic index (MI), necrosis, IHC proliferative markers Ki-67 and PHH3, and their predictive value for lung neuroendocrine tumors (NETs) aggressiveness. Materials and methods. The study used surgical and biopsy material from 64 patients with lung NETs before chemotherapy prescribing. Morphological study and IHC was performed. MI, necrosis, Ki-67 and PHH3 expression and metastatic disease and survival were estimated using nonparametric statistics. Results. Statistically significant association of necrosis severity and survival rates was found (P = 0.021). This was true for comparing patients with no necrosis in tumor tissue and extensive foci of necrosis (P = 0.023). MI appeared to be associated with metastases in lymph nodes (P = 0.003) and with distant metastatic lesions (P = 0.029). Significant, direct association of Ki-67 and MI (P < 0.001), MI and PHH3 expression (P < 0.001) was found. However, there was no significant link between Ki-67 and PHH3 rates (P = 0.240). Ki-67 didn’t show any significant association with necrosis and metastases. Also, Ki-67 rates didn’t affect the patient survival. Data on PHH3 expression and their estimation appeared to be rather contradictory. PHH3 expression rates were lower than expected and did not exceed neither Ki-67 rates, nor MI. Conclusions. MI and necrosis are reliable markers for the assessment of lung NETs aggressiveness. MI is statistically associated with metastatic lesion, while extensive necrosis – with survival rates. Ki-67 expression was significantly associated with MI. No significant association of Ki-67 and PHH3 expression, tumor’s morphological features, disease progression and prognosis was found. Contrary to our expectations, PHH3 showed no diagnostic and prognostic value in lung NETs.

International Medullary Thyroid Carcinoma Grading System: A Validated Grading System for Medullary Thyroid Carcinoma

PURPOSE Medullary thyroid carcinoma (MTC) is an aggressive neuroendocrine tumor (NET) arising from the calcitonin-producing C cells. Unlike other NETs, there is no widely accepted pathologic grading scheme. In 2020, two groups separately developed slightly different schemes (the Memorial Sloan Kettering Cancer Center and Sydney grade) on the basis of proliferative activity (mitotic index and/or Ki67 proliferative index) and tumor necrosis. Building on this work, we sought to unify and validate an internationally accepted grading scheme for MTC. PATIENTS AND METHODS Tumor tissue from 327 patients with MTC from five centers across the United States, Europe, and Australia were reviewed for mitotic activity, Ki67 proliferative index, and necrosis using uniform criteria and blinded to other clinicopathologic features. After reviewing different cutoffs, a two-tiered consensus grading system was developed. High-grade MTCs were defined as tumors with at least one of the following features: mitotic index ≥ 5 per 2 mm2, Ki67 proliferative index ≥ 5%, or tumor necrosis. RESULTS Eighty-one (24.8%) MTCs were high-grade using this scheme. In multivariate analysis, these patients demonstrated decreased overall (hazard ratio [HR] = 11.490; 95% CI, 3.118 to 32.333; P < .001), disease-specific (HR = 8.491; 95% CI, 1.461 to 49.327; P = .017), distant metastasis-free (HR = 2.489; 95% CI, 1.178 to 5.261; P = .017), and locoregional recurrence-free (HR = 2.114; 95% CI, 1.065 to 4.193; P = .032) survivals. This prognostic power was maintained in subgroup analyses of cohorts from each of the five centers. CONCLUSION This simple two-tiered international grading system is a powerful predictor of adverse outcomes in MTC. As it is based solely on morphologic assessment in conjunction with Ki67 immunohistochemistry, it brings the grading of MTCs in line with other NETs and can be readily applied in routine practice. We therefore recommend grading of MTCs on the basis of mitotic count, Ki67 proliferative index, and tumor necrosis.

Comparative Review of Malignant Melanoma and Histologically Well-Differentiated Melanocytic Neoplasm in the Oral Cavity of Dogs

Oral malignant melanoma (OMM) is the most common neoplasm of the canine oral cavity. It is characterized by its aggressive local disease as well as its high rate of lymphatic invasion and distant metastasis. OMM carries a poor prognosis, with most patients succumbing to the disease due to progression of the neoplasm. Histopathologically, OMM is characterized by significant nuclear atypia, a mitotic index of greater than 4/10 hpf, and evidence of vascular invasion or metastasis. Clinically, these lesions can become locally invasive, causing lysis of bones and severe inflammation of the surrounding soft tissue. With time, these lesions can spread to the regional lymph node and to the lungs and other organs. Prognosis can vary depending on the size of the primary tumor, regional node involvement, and distant metastatic disease; however, multiple studies report a relatively short median survival time ranging from less than 4 months to 8 months. Histologically well- differentiated melanocytic neoplasms (HWDMN) are a variant of OMM and sometimes referred to as canine oral melanocytic neoplasms of low malignant potential. Unlike OMM, patients with HWDMN have longer survival times. Histopathologically, HWDMNs have well-differentiated melanocytes with a low mitotic index of 3 or less per 10 hpf and minimal nuclear atypia. HWDMNs have better prognosis with a mean survival time of up to 34 months. This article is a comparative review of OMM and its less aggressive counterpart.

Regeneration from leaf explants of steppe cherry (Prunus fruticosa Pall)

The article represents data on morphogenesis from leaf explants of three steppe cherry genotypes, as well as the degree of somaclonal variability at in vitro and ex vitro stages, and in the field. It was revealed that a content of 6-benzylaminopurine, 4.43 μM, in combination with auxin, 0.5–0.6 μM, stimulates in the light the direct organogenesis in the tissues of the leaf base. This reaction was observed from 16.7 to 75.0% of explants, depending on the genotype. An equal 6-benzylaminopurine - auxin ratio (1: 1) led to the callus along with microshoots. Depending on the genotype, up to 30.0% of explants had such a mixed type of organogenesis. The mitotic index value in the apical leaflets differed depending on the day time. At the stage of micropropagation itself, an increase of the mitotic index was observed from 10 to 16 hours; at the stage ex vitro, no significant differences in the mitosis frequency were revealed within this time interval. No significant differences were found between the level of the mitotic index for plants obtained directly from leaves and those from buds. The mitosis passed without disturbances. No phenotypic changes in plant habit, shape and color of leaves were found.

Dynamic survival analysis of gastrointestinal stromal tumors (GISTs): a 10-year follow-up based on conditional survival

Background The prognosis of patients with gastrointestinal stromal tumors (GISTs) is generally evaluated at the time of diagnosis but does not reflect the survival dynamics of patients in the future. Therefore, the purpose of this article was to evaluate the conditional survival (CS) of Chinese patients with GISTs after radical resection. Methods This retrospective study included 451 patients who underwent radical surgery for GISTs. A Cox proportional hazard model was used to evaluate the prognostic factors of disease-free survival (DFS). The 3-year conditional DFS (CDFS3) of patients who survived for x years was expressed as CDFS3=DFS(x + 3)/DFS(x). Results The traditional 3-year DFS rate decreased gradually from 94.0% at 3 years to 77.3% at 7 years, while the CDFS3 rate increased from 94.0 to 95.2% over the survival time of the patients. In addition, classic clinicopathological prognostic factors had different effects on CDFS3, with changes observed in survival time, but these effects were only slight or moderate (|d|<0.5). Although multivariate analysis showed that age, sex, mitotic index and tumor rupture were independent risk factors for DFS at baseline, all adverse prognostic factors, except for the mitotic index, lost their predictive significance at 5 years after operation. When the Modified NIH criteria were included, the risk staging was found to be an independent risk factor for recurrence or death. Time-dependent Cox regression analysis showed that the modified NIH criteria independently affected the recurrence or death of GIST patients within 2 years after operation. Conclusion CS provides detailed dynamic survival information about Chinese patients with primary resected GISTs. The mitotic index is of great clinical significance for the monitoring and follow-up of patient populations with a high risk of tumor recurrence or death until 5 years after surgery.

Correlation of Clinical, Histopathological and Histomorphometric Features of Canine Soft Tissue Sarcomas

Soft-tissue sarcomas (STS) represent a heterogeneous group of tumours with similar histological characteristics and biological behaviour. This study aimed to describe the correlation between clinical, histopathological and histomorphometric features of STS in dogs. Medical records were reviewed to identify all dogs in which an STS was diagnosed between 2006-2017. Thirty cases were included, and tumour samples and medical records were recovered. Most of the dogs were mixed breed (40%) and 80% of the STS were located in the subcutaneous connective tissue. Histopathological classification showed that undifferentiated sarcoma (17%) and peripheral nerve sheath tumour (30%) were the most common STS. Grade I STS were obtained in 50% of cases (15/30), and grade II or III tumours compromised 43% (13/30) and 7% (2/30) respectively. The mitotic index ranged from zero to 26 (5.8 ± 7.5). Increased nucleus:cytoplasm ratio was moderately associated with higher tumour grade (p = 0.05; rS = 0.361) and mitotic index (p = 0.05; rS = 0.355), while the number of microvessels was positively correlated with degree of differentiation (p = 0.05; rS = 0.362) and nuclear pleomorphism (p = 0.036; rS = 0.384). Histomorphometry proved to be useful in the evaluation of STS, representing an additional tool correlated with well-established prognostic factors (histopathological grade, degree of differentiation, nuclear pleomorphism).