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

Potential Diagnostic and Prognostic Biomarkers for Adenovirus Respiratory Infection in Children and Young Adults

Human Adenoviruses (HAdV) are known to be potentially associated with strong inflammatory responses and morbidity in pediatric patients. Although most of the primary infections are self-limiting, the severity of clinical presentation, the elevation of the white blood cell count and inflammatory markers often mimic a bacterial infection and lead to an inappropriate use of antibiotics. In infections caused by HAdV, rapid antigen detection kits are advisable but not employed routinely; costs and feasibility of rapid syndromic molecular diagnosis may limit its use in the in-hospital setting; lymphocyte cultures and two-sampled serology are time consuming and impractical when considering the use of antibiotics. In this review, we aim to describe the principal diagnostic tools and the immune response in HAdV infections and evaluate whether markers based on the response of the host may help early recognition of HAdV and avoid inappropriate antimicrobial prescriptions in acute airway infections.

A NEW METHODOLOGY FOR DIAGNOSIS OF FANCONI ANEMIA BASED ON BIOLOGICAL DOSIMETRY

Fanconi Anemia (FA) is a syndrome associated with chromosomal fragility. Current laboratory tests to diagnose this disease are based on the scoring of chromosomal aberrations induced in peripheral blood lymphocytes by clastogenic chemical agents, mainly: diepoxybutane (DEB) or mitomycin C (MMC). This study evaluated an alternative test for the diagnosis of FA, in which ionizing radiation replaces DEB/MMC. Two groups were studied: normal and DEB-sensitive individuals. From each individual, samples of peripheral blood were irradiated using an electron linear accelerator. Following lymphocyte cultures, and slide preparation, metaphases were scored based on the same methodology for biological dosimetry, according to recommendations of the International Atomic Energy Agency. Our results emphasized a pattern of distribution of dicentrics, fragments, as well as abnormal chromosomal arrangements. The methodology of analysis here proposed permitted to distinguish normal from DEB-sensitive subjects.

Chromosome Aberrations Produced by Mestranol in Human Lymphocyte Cultures

In this investigation, the genotoxic properties of mestranol were examined in vitro. Human lymphocyte cultures were exposed for 72 h to mestranol at concentrations of 7.5, 15 and 30 µg/g. The genotoxic effects of the chemosterilant were assessed by numerical and structural chromosome aberrations. Mestranol induced certain genotoxic effects in human lymphocytes. There was a dose-dependent significant (p<0.01) increase in the number of numerical aberrations in comparison to the control, but without significant differences (p>0.05) between the doses applied. Further, structural aberrations increased significantly (p<0.01) in the presence of mestranol, being most frequent in cultures exposed to the highest mestranol dose. The frequency of Robertsonian translocations increased significantly only in cultures treated with mestranol at concentration of 30 µg/g in comparison both with the control (p<0.01) and the lowest chemosterilant dose (p<0.01). There were significant differences (p<0.01) in the levels of chromosome gaps and fragments compared to Robertsonian translocations, whilst the frequencies between gaps and fragments were not significantly different (p>0.05).

EFFECTS OF FEMALE BLOOD AUTOSERUM ON ALLOGENIC INTERACTIONS IN SHORT-TERM LYMPHOCYTE CULTURES OF PARENTS HAVING CHILDREN WITH CONOTRUNCAL HEART MALFORMATIONS

Highlights The findings of this original study ensure the detection of violations in the humoral regulation of the maternal immune interactions with semiallogeneic fetus, considered as a risk factor for developing sporadic conotruncal heart malformations in the next generation.Aim To study the role of female autoserum blood in limiting allogeneic interactions in short-term lymphocyte cultures of parents having children with conotruncal heart malformations.Methods 21 married couples (the study group) with children suffering from conotrucnal heart malformations (Tetralogy of Fallot) without chromosomal diseases were examined. The control group consisted of 21 families with three or more healthy children. The immune response in a mixed lymphocyte culture of parents was assessed by the increase in HLA-DR expression in the mixed culture with respect to spontaneous lymphocyte cultures. Primary staining of female and male lymphocytes with monoclonal antibodies to CD45, conjugated with various fluorescent dyes (PC-5 and PC-7), allowed assessing the immune response of female lymphocytes to male and vice versa.Results The effects of female autoserum on the mixed lymphocyte culture of parents were assessed. The obtained results reported that the birth of children with conotruncal heart malformations is associated with the interfering effect of female autoserum on HLA-DR expression on subpopulations of female lymphocytes (CD3+, HLA-DR+) and the activating effect on subpopulations of female lymphocytes (CD3-, HLA-DR+). The observed role of female autoserum in the study group may be associated with the absence of HLA-DR-blocking autoantibodies and high synthesis of cytokines by T2 and T3 helper lymphocytes.Conclusion The effects of female autoserum on allogeneic lymphocyte interactions of parents may be observed in short-term mixed lymphocyte cultures. The evaluation of the activating and interfering effects ensures timely identification of any violations in the humoral regulation of the maternal immune interactions with the HLA semiallogenic fetus, considered as a risk factor for developing sporadic conotruncal heart malformations in the next generation.

Induction of lymphocyte apoptosis in healthy individuals and patients with rheumatoid arthritis under “cellular neighborhood” in vitro

The aim of the study was to investigate the features of T-lymphocyte apoptosis induced by components of autologous apoptotic cultures in vitro in norm and rheumatoid arthritis in the context of «cellular neighborhood».Materials and methods. Subjects of the study were blood samples of patients with rheumatoid arthritis (RA) and healthy women of comparable age. Developed protocol allowed to differentially evaluate the parameters of proliferation, early and late stages of apoptosis in the «primary» (CFSE-) and «secondary» (CFSE+) induced apoptotic T-lymphocyte cultures. It was estimated the effect of cellular and humoral components of unstimulated, anti-CD3- and dexamethasone-stimulated cells under the conditions of overcrowding and depleted culture media on autologous lymphocytes, cultured under physiological conditions, in norm and RA.Results. Comparative qualitative analysis revealed the features of the processes of T-lymphocyte apoptosis in norm and pathology. Also, the parameters of early and late stages of apoptosis of a «primary» induced culture and «secondary» induced cells after transferring the cellular and humoral components of apoptotic cultures did not differ significantly either initially or during culturing in both investigated groups. But it was a significant increase in the amount of living T-cells in «primary»-induced unstimulated and dexamethasone-stimulated RA patients’ cultures compared to similar donors’ cultures.Conclusion. There was no difference between stimulated with anti-CD3 antibodies cells and the «secondary» induced cultures. Taking into account the absence of significant differences in the parameters of activation apoptosis, the increased number of living cells in RA patients’ cultures relative to donors’ is evidence of contribution of non-autonomous apoptosis effects to cellular homeostasis in RA.

Cytogenetic alterations induced by flupyradifurone, a new butenolide insecticide, in human lymphocytes

Flupyradifurone (FPD), a member of the new class of butenolide insecticides, acts on nicotinic acetylcholine receptors. Studies on genotoxic and carcinogenic effects of FPD are very limited. This is the first study to investigate the cytotoxic and genotoxic effects of FPD and its metabolites on human lymphocyte cultures with or without a metabolic activation system (S9 mix) using chromosomal aberration (CA) and micronucleus (MN) tests. The cultures were treated with 85, 170, and 340 µg/ml of FPD in the presence (3 h treatment) and absence (48 h treatment) of S9 mix. Dimethyl sulfoxide (DMSO) was used as a solvent control. Statistically significant decreases were detected at the medium and highest concentrations for 48 h treatments while decreases in mitotic index (MI) in the presence of the S9 mix were found statistically significant at all FPD concentrations tested when compared with the solvent control. FPD also decreased the nuclear division index (NDI) at the highest concentration (340 µg/ml) in the absence of S9 mix. When compared with the solvent control, increases in CA frequencies were significant at the medium and highest concentrations. Significantly increased MN frequency was only found at the highest FPD concentration in cultures without S9 mix compared with the solvent control while increases in the MN frequencies in the presence of S9 mix were statistically significant at all FPD concentrations. The results of the present study indicate that FPD and its metabolites can show cytotoxic and genotoxic effects in human lymphocytes. More genotoxicity studies are necessary to make a possible risk assessment in humans.