Damage In Human Lymphocytes Research Articles

A study on Amygdalin's Genotoxicological Safety and Modulatory Activity in Human Peripheral Lymphocytes in vitro.

Amygdalin (AMY), a plant secondary metabolite containing nitrile, is a major component of the seeds of Rosaceae family plants. It is known that this compound has many pharmacological activities such as cancer prevention, antipyretic, and cough suppressant. In this study, the genotoxic and modulatory effects of amygdalin were assessed by chromosomal aberration (CA), sister chromatid exchange (SCE), and cytokinesis-block micronucleus assay (CBMN) assays using human peripheral lymphocytes (HPLs) in the absence and presence of metabolic activator (S9 mix). Lymphocytes were exposed to various concentrations of amygdalin (0.86, 1.72, 3.43, 6.86, and 13.75 μg/mL) alone and in combination with mitomycin-C (MMC, 0.20 μg/mL) or cyclophosphamide (CP, 12 μg/mL). The mitotic index (MI), replication index (RI), cytokinesis-block proliferation index (CBPI), and cytostasis were also evaluated to determine cytotoxicity. Amygdalin alone did not exhibit genotoxic and cytotoxic effects at all the tested concentrations both in the absence and presence of the S9 mix. In contrast, amygdalin significantly reduced the frequencies of CA (especially at 48 h treatments), SCE, and MN (except 0.86 μg/mL in pre- and simultaneous treatment) induced by MMC in all the tested concentrations and treatment protocols. It has also considerably decreased CP-induced CA and SCE frequencies at all the concentrations (except 0.86 μg/mL) in simultaneous treatment. This study demonstrated that amygdalin alone was not genotoxic, on the contrary, it has revealed modulatory effects against chemotherapy agents that induced genomic damage in human lymphocytes, suggesting its chemopreventive potential. This article is protected by copyright. All rights reserved.

Methanolic fraction of Cassia fistula L. bark exhibits potential to combat oxidative stress and possess antiproliferative activity

ABSTRACT Cassia fistula L. is well known for its traditional medicinal properties as an anti-inflammatory, hepatoprotective, antifungal, antibacterial, antimutagenic, and wound healing agent. The aim of the present study was to determine antioxidant, genoprotective, and cytotoxic potential of different fractions of C. fistula bark including hexane (CaMH), chloroform (CaMC), ethyl acetate (CaME), and methanol (CaMM). Among all the fractions studied, CaMM exhibited maximal radical scavenging activity in antioxidant DPPH assay, Superoxide anion radical scavenging assay and nitric oxide radical scavenging assay displayed an IC50 value of 18.95, 29.41, and 13.38 µg/ml, respectively. CaMM fraction possessed the highest phenolic (130.37 mg gallic acid equivalent/g dry weight of extract) and flavonoid (36.96 mg rutin equivalent/g dry weight of fraction) content. Data demonstrated significant positive correlation between polyphenol levels and radical scavenging activity. Single cell gel electrophoresis (Comet assay) exhibited genoprotective potential of C. fistula bark fractions against DNA damage induced by hydrogen peroxide (H2O2) in human lymphocytes. CaMM fraction displayed highest protective ability against H2O2 induced-toxicity as evidenced by significant decrease in % tail DNA content from 30 to 7% at highest concentration (200 µg/ml). CaMM was found to be rich in catechin, gallic acid, chlorogenic acid, and kaempferol. The phenolic content and antioxidant ability of the fractions was markedly negatively correlated with H2O2- induced DNA damage in human lymphocytes. Cytotoxic potential was evaluated against dermal epidermoid carcinoma (A431), pancreatic (MIA PaCa-2) and brain glioblastoma (LN-18) cancer cell lines using MTT assay. Results showed that C. fistula bark fractions possessed highest toxicity against the skin carcinoma cells. CaMM fraction reduced over 50% cell growth at the concentration of 76.72 µg/ml in A431 cells. These findings suggest that fractions of C. fistula bark exhibit potential to be considered as therapeutic agents in various carcinomas.

Ratio-dependent effects of photoactivated hypericin and manumycin A on their genotoxic and mutagenic potential

Natural compounds originated from plants and microorganisms and their combinations are currently being investigated as a possible treatment for several diseases including cancer. Hypericin (photodynamically-active pigment from Hypericum perforatum L.) and manumycin A (inhibitor of farnesyltransferase from Streptomyces parvulus) belong to the chemicals potentially applicable in clinical practice. In this study we evaluated potential cytotoxic (via trypan blue exclusion test), genotoxic (via DNA-topology and comet assays), and mutagenic effects (via bacterial reverse mutation test) of these compounds and their combinations considering the molecular mechanism of their action in cell-free and cellular systems. Our results did not reveal neither cytotoxic nor mutagenic activities of tested compounds and their combinations. Regarding the genotoxic potential, no damage of plasmid DNA in cell-free system was detected. On the other hand, photoactivated hypericin and manumycin A were able to induce primary DNA damage in human lymphocytes analyzed by comet assay. The possible antagonistic interactions between these two metabolites were estimated using SynergyFinder software analysis and experimental data obtained from comet assay. Our findings indicate that not only the presence of substances, but also their ratio plays an important role in resulting effects of the combined treatment in cellular system.

Protective Effects of Arbutus unedo L. Honey in the Alleviation of Irinotecan-Induced Cytogenetic Damage in Human Lymphocytes-An In Vitro Study.

Strawberry tree (Arbutus unedo L.) honey (STH) has been used since ancient times as a folk medicine remedy, especially in certain Mediterranean countries. This honey, rich in phenolic content, is well recognized for its antioxidant, anti-inflammatory, and antimicrobial activities, and is used for the treatment of skin lesions as well as gastrointestinal and respiratory disorders. This study investigated whether STH alleviates genome damage in human peripheral blood lymphocytes produced by the cytotoxic drug irinotecan. The phenolic profile of STH was previously estimated by ultra-high-performance liquid chromatography coupled to a linear ion trap-Orbitrap hybrid mass spectrometer. The effects of STH were evaluated at three concentrations (1×, 5×, and 10×), based on the daily consumption of the honey by an adult person. After 2 h of in vitro exposure, standard lymphocyte cultures for the analysis of chromosome aberrations and the cytokinesis-block micronucleus cytome assay were established. Our results demonstrate that STH offered remarkable geno- and cytoprotection when administered with irinotecan. These findings are relevant for drawing preliminary conclusions regarding the in vitro safety of the tested honey. However, further studies are needed with the application of more complex experimental models.

Effect of curcumin on γ-ray-induced cell response.

The purpose of the present study is to evaluate the effect of curcumin as a natural compound against radiation induced γ-foci and stable chromosome aberrations. Whole blood samples form three human volunteers were pretreated with curcumin at different concentrations (0.5, 10, 20 and 100μg/ml). After 1-hour incubation, the lymphocytes were exposed to γ-rays (0.05, 0.5, 1 and 2Gy). Radiation induced changes in cells were quantified using γ-H2AX/53BP1 assay and FISH analysis. Our results have shown that curcumin significantly reduced the frequency of both γ-foci and translocations. We found concentration-dependent increase of curcumin protective effect on γ-H2AX/53BP1 foci formation at all radiation doses. Concerning the translocations, after 0.05 and 0.5Gy γ-rays the values of genomic frequencies are comparable within each dose and we did not observe any impact of curcumin. The most protective effect after 1Gy exposure was found at 100μg/ml curcumin. At 2Gy irradiation, the maximum protection was achieved at 0.5 and 10μg/ml of curcumin. Concentrations of 20 and 100μg/ml also prevent lymphocytes but to less extent. Our in vitro study indicates radioprotective efficacy of curcumin against γ-ray induced damages in human lymphocytes. This observation suggests that curcumin may play a role to protect patients undergoing radiological procedures.

Pharmacology and early ADMET data of corallopyronin A, a natural product with macrofilaricidal anti-wolbachial activity in filarial nematodes

Corallopyronin A (CorA), a natural product antibiotic of Corallococcus coralloides, inhibits the bacterial DNA-dependent RNA polymerase. It is active against the essential Wolbachia endobacteria of filarial nematodes, preventing development, causing sterility and killing adult worms. CorA is being developed to treat the neglected tropical diseases onchocerciasis and lymphatic filariasis caused by Wolbachia-containing filariae. For this, we have completed standard Absorption, Distribution, Metabolism, Excretion and Toxicity (ADMET) studies. In Caco-2 assays, CorA had good adsorption values, predicting good transport from the intestines, but may be subject to active efflux. In fed-state simulated human intestinal fluid (pH 5.0), CorA half-life was >139 minutes, equivalent to the stability in buffer (pH 7.4). CorA plasma-stability was >240 minutes, with plasma protein binding >98% in human, mouse, rat, dog, mini-pig and monkey plasma. Clearance in human and dog liver microsomes was low (35.2 and 42 µl/min/mg, respectively). CorA was mainly metabolized via phase I reactions, i.e., oxidation, and to a minimal extent via phase II reactions. In contrast to rifampicin, CorA does not induce CYP3A4 resulting in a lower drug-drug-interaction potential. Apart from inhibition of CYP2C9, no impact of CorA on enzymes of the CYP450 system was detected. Off-target profiling resulted in three hits (inhibition/activation) for the A3 and PPARγ receptors and COX1 enzyme; thus, potential drug-drug interactions could occur with antidiabetic medications, COX2 inhibitors, angiotensin AT1 receptor antagonists, vitamin K-antagonists, and antidepressants. In vivo pharmacokinetic studies in Mongolian gerbils and rats demonstrated excellent intraperitoneal and oral bioavailability (100%) with fast absorption and high distribution in plasma. No significant hERG inhibition was detected and no phototoxicity was seen. CorA did not induce gene mutations in bacteria (Ames test) nor chromosomal damage in human lymphocytes (micronucleus test). Thus, CorA possesses an acceptable in vitro early ADMET profile; supported by previous in vivo experiments in mice, rats and Mongolian gerbils in which all animals tolerated CorA daily administration for 7-28 days. The non-GLP package will guide selection and planning of regulatory-conform GLP models prior to a first-into-human study.

Differences in sensitivity of human lymphocytes and fish lymphocytes to polyvinyl chloride microplastic toxicity.

Polyvinyl chloride (PVC) microplastics are emerging contaminants affecting biological wastewater treatment processes. So far, the toxicological investigation of PVC microplastics usually focused on the anaerobic and denitrifying bacteria. It seems that the primary lymphocytes isolated from peripheral blood are more sensitive than most other organ cell types in vitro; therefore, the aim of this study was to assess the cytotoxicity of PVC microplastic on human and fish blood lymphocytes as a useful ex vivo model for accelerated human toxicity studies. Using biochemical analyses, we showed human lymphocytes are more sensitive to toxic effects of PVC microplastic than fish lymphocytes. Our result showed that addition of PVC microplastic at 24, 48, and 96μg/ml for 3h to human blood lymphocytes induced cytotoxicity. The PVC microplastic-induced cytotoxicity on human blood lymphocytes was associated with intracellular reactive oxygen species (ROS) formation, lysosomal membrane injury, mitochondrial membrane potential (MMP) collapse, depletion of glutathione, and lipid peroxidation. According to our results, PVC microplastic particles induce oxidative stress and organelle damage in human lymphocytes, while these significant alterations in toxicity parameters in PVC microplastic-treated fish lymphocytes were not observed. Finally, our findings suggest that human lymphocytes are more sensitive to PVC microplastic toxicity compared with fish lymphocytes.

Cytogenetic Damage of Human Lymphocytes in Humanized Mice Exposed to Neutrons and X Rays 24 h After Exposure

Detonation of an improvised nuclear device highlights the need to understand the risk of mixed radiation exposure as prompt radiation exposure could produce significant neutron and gamma exposures. Although the neutron component may be a relatively small percentage of the total absorbed dose, the large relative biological effectiveness (RBE) can induce larger biological DNA damage and cell killing. The objective of this study was to use a hematopoietically humanized mouse model to measure chromosomal DNA damage in human lymphocytes 24 h after in vivo exposure to neutrons (0.3 Gy) and X rays (1 Gy). The human dicentric and cytokinesis-block micronucleus assays were performed to measure chromosomal aberrations in human lymphocytes in vivo from the blood and spleen, respectively. The mBAND assay based on fluorescent in situ hybridization labeling was used to detect neutron-induced chromosome 1 inversions in the blood lymphocytes of the neutron-irradiated mice. Cytogenetics endpoints, dicentrics and micronuclei showed that there was no significant difference in yields between the 2 irradiation types at the doses tested, indicating that neutron-induced chromosomal DNA damage in vivo was more biologically effective (RBE ∼3.3) compared to X rays. The mBAND assay, which is considered a specific biomarker of high-LET neutron exposure, confirmed the presence of clustered DNA damage in the neutron-irradiated mice but not in the X-irradiated mice, 24 h after exposure.

Linalool reverses benzene-induced cytotoxicity, oxidative stress and lysosomal/mitochondrial damages in human lymphocytes

Benzene exposure results in bone marrow suppression, leading to a decrease in the number of circulating white blood cells, an increased risk of chronic lymphocytic leukemia, acute myeloid leukemia and aplastic anemia. Since the mechanism of induction of benzene toxicity is due to active metabolites through cytochrome p450 enzymes and production of reactive oxygen species (ROS), we hypothesized that natural compound such linalool with anti-inflammatory/antioxidant properties could be effective in reducing its toxicity. Lymphocytes isolated from healthy individuals were simultaneously cotreated with different concentrations of LIN (10, 25 and 50 µM) and benzene (50 µM) for 4 h at 37 °C. After incubation, the toxicity parameters such cytotoxicity, ROS formation, lysosomal membrane integrity, mitochondria membrane potential (ΔΨm) collapse, oxidized/reduced glutathione (GSH/GSSG) and malondialdehyde (MDA) were analyzed using biochemical and flow cytometry evaluations. Our data showed that benzene (50 µM) induced a significant increase in cytotoxicity, ROS formation, mitochondrial membrane potential (MMP) collapse, lipid peroxidation and oxidative stress while LIN with antioxidant potential reversed the toxic effects of benzene on isolated human lymphocytes. Our results suggest that LIN reduces and reverses benzene-induced cytotoxicity, oxidative stress and lysosomal/mitochondrial damages in human lymphocyte. This study demonstrated that cotreatment of LIN with benzene can reduce several parameters indicative of oxidative stress. As such, LIN could represent a potential therapeutic agent in reducing certain aspects of benzene-induced toxicity.

Intercomparison of Conventional and QuickScan Dicentric Scoring for the Validation of Individual Biodosimetry Analysis in Taiwan

Purpose The dicentric chromosome assay (DCA), the gold standard for radiation biodosimetry, evaluates an individual absorbed radiation dose by the analysis of DNA damage in human lymphocytes. The conventional (C-DCA) and QuickScan (QS-DCA) scoring methods are sensitive for estimating radiation dose. The Biodosimetry Laboratory at Institute of Nuclear Energy Research (INER), Taiwan, participated in intercomparison exercises conducted by Health Canada (HC) in 2014, 2015 and 2018 to validate the laboratory’s accuracy and performance. Material and Methods Blood samples for the conventional dose response curve for Taiwan were irradiated with 0, 0.25, 0.5, 1, 2, 3, 4 and 5 Gy. Ten blind blood samples were provided by HC. Either or both of two methods of conventional (C) or QuickScan (QS) scoring could be chosen for the HC’s intercomparison. For C-DCA triage scoring, only cells with 46 centromeres were counted and each scorer recorded the number of dicentrics in the first 50 metaphases or stopped scoring when 30 dicentrics were reached. Scorers also recorded how much time it took to analyze 10, 20, and 50 cells. Subsequently, the data were entered into the Dose Estimate software (DoseEstimate_v5.1) and dose estimates were calculated. With QS-DCA scoring, a minimum of 50 metaphase cells (or 30 dicentrics) were scored in apparently complete metaphases without verification of exactly 46 centromeres. Results For the blinded blood samples irradiated at HC and shipped to INER, the mean absolute deviation (MAD) derived after scoring 50 cells for C-DCA and QS-DCA was <0.5 Gy for all three intercomparisons, meeting the criteria for acceptance. Conclusion The results indicated that the Biodosimetry Laboratory at INER can provide reliable dose estimates in the case of a large-scale radiation accident.

Mitochondrial, lysosomal and DNA damages induced by acrylamide attenuate by ellagic acid in human lymphocyte.

Acrylamide (AA), is an important contaminant formed during food processing under high temperature. Due to its potential neurotoxicity, reproductive toxicity, hepatotoxicity, immunotoxicity, genotoxicity and carcinogenicity effects, this food contaminant has been recognized as a human health concern. Previous studies showed that acrylamide-induced toxicity is associated with active metabolite of acrylamide by cytochrome P450 enzyme, oxidative stress, mitochondrial dysfunction and DNA damage. In the current study, we investigated the role of oxidative stress in acrylamide's genotoxicity and therapeutic potential role of ellagic acid (EA) in human lymphocytes. Human lymphocytes were simultaneously treated with different concentrations of EA (10, 25 and 50 μM) and acrylamide (50 μM) for 4 h at 37°C. After 4 hours of incubation, the toxicity parameters such cytotoxicity, ROS formation, oxidized/reduced glutathione (GSH/GSSG) content, malondialdehyde (MDA) level, lysosomal membrane integrity, mitochondria membrane potential (ΔΨm) collapse and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were analyzed using biochemical and flow cytometry evaluations. It has been found that acrylamide (50 μM) significantly increased cytotoxicity, ROS formation, GSH oxidation, lipid peroxidation, MMP collapse, lysosomal and DNA damage in human lymphocytes. On the other hand, cotreatment with EA (25 and 50 μM) inhibited AA-induced oxidative stress which subsequently led to decreasing of the cytotoxicity, GSH oxidation, lipid peroxidation, MMP collapse, lysosomal and DNA damage. Together, these results suggest that probably the co-exposure of EA with foods containing acrylamide could decrease mitochondrial, lysosomal and DNA damages, and oxidative stress induced by acrylamide in human body.

Protective effects of (-)-epigallocatechin gallate and curcumin against acrylamide toxicity

Human lymphocytes as well as zebrafish embryos were used to study the toxicity of acrylamide, and the intervention effects of the dietary antioxidants (-)-epigallocatechin gallate and curcumin have been examined. The alkaline comet assay revealed that acrylamide (1, 5, 10 mmol/L) had dose-response effects on basic DNA damage and oxidative DNA damage in human lymphocytes. Exposure of zebrafish embryos to acrylamide resulted in decrease of hatching rate, and in increases of mortality and teratogenicity. (-)-Epigallocatechin gallate and curcumin reduced DNA damage and protected zebrafish embryos in the growth and development.

Ex vivo/in vitro effects of aspirin and ibuprofen, bulk and nano forms, in peripheral lymphocytes of prostate cancer patients and healthy individuals

Inhibiting inflammatory processes or eliminating inflammation represents a logical role in the suppression and treatment strategy of cancer. Several studies have shown that anti-inflammatory drugs (NSAIDs) act as anticancer agents while reducing metastases and mortality rate. NSAIDs are seriously limited by their side effects and toxicity, which can become cumulative with their long-term administration for chemoprevention. In the current ex vivo / in vitro study, the genotoxicity mechanisms of NSAIDS in bulk and nanoparticle forms allowed a strategy to prevent and minimise the damage in human lymphocytes. When compared to their bulk forms, acetylsalicylic acid (Aspirin) nano and ibuprofen nano (IBU N), both NSAIDs in 500 μg/mL concentration significantly decreased DNA damage measured by alkaline comet assay. Micronuclei (MNi) frequency also decreased after ASP N (500 μg/mL), ASP B (500 μg/mL) and IBU N (200 μg/mL) in prostate cancer patients and healthy individuals, however, the ibuprofen bulk (200 μg/mL) showed a significant increase in MNi formation in lymphocytes from healthy and prostate cancer patients when compared to the respective untreated lymphocytes. These findings suggest that a reduction in particle size had an impact on the reactivity of the drug, further emphasising the potential of nanoparticles to improve the current treatment options.

Primary screening for the toxicity of marine cyanobacteria Lyngbya bouillonii (Cyanophyceae: Oscillatoriales) recorded for the first time from Indian Ocean

Climate change has triggered an increased incidence of cyanobacterial blooms in the marine ecosystem. While most bloom-forming species cause widespread algal blooms, Lyngbya bouillonii, a benthic marine cyanobacteria forms localized blooms. This cyanobacteria, reported only from the tropical Pacific Ocean, has gained immense biomedical interest owing to an array of bioactive secondary metabolites it produces. While studying the occurrence and distribution of marine cyanobacteria across Andaman Nicobar Islands we encountered this species forming tenacious mats, attached to corals and quite often forming extensive localized blooms. Accordingly, L. bouillonii is being reported for the first time from the Indian Ocean. The symbiotic, snapping shrimp Alpheus frontalis was found to coexist within these cyanobacterial mats. Our initial toxicity studies showed L. bouillonii to possess toxins, causing mortality of brine shrimps with a LC50 of 0.8 mg/ml and causing DNA damage in human lymphocytes in vitro at concentrations as low as 1 ng/ml of crude cyanobacterial extract. Preliminary investigation revealed the presence of chlorinated fatty acid amides, fatty acid esters and benzazepine derivatives in the extract. Furthermore, how this cyanobacteria initially reached Indian waters and whether this is a bio-invasion is still being ascertained.

Protective effects of olive oil phenolics oleuropein and hydroxytyrosol against hydrogen peroxide-induced DNA damage in human peripheral lymphocytes.

This study investigates antioxidant capacity and protective effects of phenolic compounds oleuropein (OLP) and hydroxytyrosol (HT), present in olive oil and olive leaves, against H2O2-induced DNA damage in human peripheral lymphocytes. Antioxidant potency was determined using the measurement of radical-scavenging activity (ABTS∙+ assay), ferric reducing power (FRAP assay) and cupric reducing antioxidant capacity (CUPRAC assay). Both substances were found to be potent antioxidant agents due to their free radical-scavenging activities. Antigenotoxic effects of oleuropein and hydroxytyrosol against H2O2-induced damage in human lymphocytes were evaluated in vitro by alkaline comet assay. At tested concentrations (1, 5, 10 µmol L-1), oleuropein and hydroxytyrosol did not induce a significant increase of primary DNA damage in comparison with the negative control. Pretreatment of human lymphocytes with each of the substances for 120 min produced a dose-dependent reduction of primary DNA damage in the tested cell type. Hydroxytyrosol showed a better protective effect against H2O2-induced DNA breaks than oleuropein which could be associated with their free radical-scavenging efficacy.

Inhibitory effect of carnosine and anserine on DNA oxidative damage induced by Fe 2+, Cu 2+ and H 2O 2 in lymphocytes

The effects of carnosine and anserine on oxidative DNA damage in human lymphocytes induced by Fe 2+, Cu 2+ or H 2O 2 were investigated using single cell gel electrophoresis (comet assay). Carnosine and anserine induced a slight DNA damage in human lymphocytes after incubation with cells for 30 min. Carnosine and anserine caused a tail DNA % from 4.7% to 7.3% and 6.1% to 10.0% respectively, at a concentration of 5-100 mM. The cell viability of carnosine and anserine to human lymphocytes was more than 85%. Fe 2+ and Cu 2+ caused a marked cytotoxicity and DNA damage in human lymphocytes with a concentration dependent manner. At a concentration of 100 μM, carnosine and anserine possessed 60-70% inhibitory effect on DNA damage in human lymphocytes when they reacted with Fe 2+ or Cu 2+ for 30 min before incubated with human lymphocytes. Fe 2+, Cu 2+, and H 2O 2-induced DNA damage in human lymphocytes were inhibited by carnosine and anserine in a concentration dependent manner (5-100 μM). However, the inhibitory effect of carnosine and anserine on oxidative DNA damage in human lymphocyte was 46.4% and 49.3%, respectively, when reacted simultaneously with H 2O 2. Carnosine and anserine possessed more marked inhibitory effect on oxidative DNA damage in human lymphocyte induced by Fe 2+ and Cu 2+ than that induced by H 2O 2. This result might be due to carnosine and anserine did not possess significantly scavenging effect on H 2O 2, thus, H 2O 2 entered the cell and caused DNA damage.

Aberrant Function of the C-Terminal Tail of HIST1H1E Accelerates Cellular Senescence and Causes Premature Aging

Histones mediate dynamic packaging of nuclear DNA in chromatin, a process that is precisely controlled to guarantee efficient compaction of the genome and proper chromosomal segregation during cell division and to accomplish DNA replication, transcription, and repair. Due to the important structural and regulatory roles played by histones, it is not surprising that histone functional dysregulation or aberrant levels of histones can have severe consequences for multiple cellular processes and ultimately might affect development or contribute to cell transformation. Recently, germline frameshift mutations involving the C-terminal tail of HIST1H1E, which is a widely expressed member of the linker histone family and facilitates higher-order chromatin folding, have been causally linked to an as-yet poorly defined syndrome that includes intellectual disability. We report that these mutations result in stable proteins that reside in the nucleus, bind to chromatin, disrupt proper compaction of DNA, and are associated with a specific methylation pattern. Cells expressing these mutant proteins have a dramatically reduced proliferation rate and competence, hardly enter into the S phase, and undergo accelerated senescence. Remarkably, clinical assessment of a relatively large cohort of subjects sharing these mutations revealed a premature aging phenotype as a previously unrecognized feature of the disorder. Our findings identify a direct link between aberrant chromatin remodeling, cellular senescence, and accelerated aging.

Protective effect of extracts of Teucrium Polium and Rumex Crispus against cyclophosphamide-induced genotoxic damage in human lymphocytes

Teucrium polium (T. polium) and Rumex crispus (R. crispus) are plant species that grow widely in Anatolia and are thought to have healing effects for many diseases. In this study plant extracts are suggested as alternative agents in repairing cellular damage by using sister chromatid exchange (SCE), micronucleus (MN), mitotic index (MI), replication index (RI) and nuclear abnormalities (NAs), against the genotoxicity of cyclophosphamide (CP) in the human lymphocyte cells. 8 experimental groups were formed in the study. The cell culture medium was supplemented with 0.16 g/ml CP and the cells were treated with 50, 100 and 250 M T. polium and R. crispus extracts in the presence and absence of CP. As a result, CP significantly decreased MI frequency while increasing SCE, MN and NAs frequencies in cells. 100 M T. polium plus CP decreased SCEs when compared with CP alone. In addition, MN frequency was significantly decreased in 100 M T. poliumplus CP and 250 M R. crispus plus CP combine groups. Our results suggest that these plant extracts are not genetically damaging and have improving effects at these doses.

Phorate triggers oxidative stress and mitochondrial dysfunction to enhance micronuclei generation and DNA damage in human lymphocytes.

Herein, we studied phorate for its toxicological effects in human lymphocytes. Phorate treatment for 3 h has induced significant increase in the lymphocytic DNA damage. Compared to control, comet data from highest concentration of phorate (1000 µM) showed 8.03-fold increase in the Olive tail moment (OTM). Cytokinesis blocked micronucleus (CBMN) assay revealed 6.4-fold increase in binucleated micronucleated (BNMN) cells following the exposure with phorate (200 µM) for 24 h. The nuclear division index (NDI) in phorate (200 µM) treated cells reduced to 1.8 vis-à-vis control cells showed NDI of 1.94. Comparative to untreated control, 60.43% greater DCF fluorescence was quantitated in lymphocytes treated with phorate (500 µM), affirming reactive oxygen species (ROS) generation and oxidative stress. Flow cytometric data of phorate (200 µM) treated lymphocytes showed 81.77% decline in the fluorescence of rhodamine 123 (Rh123) dye, confirming the perturbation of mitochondrial membrane potential (ΔΨm). Calf thymus DNA (ct-DNA) treated with phorate (1000 µM) exhibited 2.3-fold higher 8-Hydroxy-2′-deoxyguanosine (8-oxodG) DNA adduct formation, signified the oxidative DNA damage. The alkaline unwinding assay revealed 4.0 and 6.5 ct-DNA strand breaks when treated to phorate and phorate-Cu (II) complex. Overall, the data unequivocally suggests the cyto- and genotoxic potential of phorate in human lymphocytes, which may induce comparable toxicological consequences in persons occupationally or non-occupationally exposed to insecticide phorate.

Eggplant fruits protect against DNA damage and mutations

Eggplant belongs to the Solanaceae family, and it has an important antioxidant capability that has been shown to counteract oxidation, which is harmful to health and many diseases. In this present study, we evaluated the antigenotoxic effects of six eggplants ((Solanum aculeatissimum Jacq. ‘Ma-khuea-lueang’; ML), (Solanum aculeatissimum Jacq. ‘Ma-khuea-pro’; MP), (Solanum aculeatissimum Jacq. ‘Ma-khuea-sawoei’; MS), (Solanum melongena Linn. ‘Ma-khuea-khai-tao’; MKT), (Solanum melongena Linn. ‘Ma-khuea-muang klom’; MM) and (Solanum torvum Sw. ‘Ma-khuea-phuang’; MPH)) against urethane-induced somatic mutation and recombination test (SMART) in Drosophila melanogaster and hydrogen peroxide-induced oxidative DNA damage in human lymphocytes. First, we determined all of the eggplant extracts of their antioxidant properties including radical scavenging activities, reducing antioxidant power and total phenolic contents, surprisingly ML extract showed the highest level of activity. In SMART, larvae were fed with each lyophilized eggplant. The results revealed that no sample was mutagenic. Interestingly, we found that all six eggplants had a potent inhibitory effect against urethane-induced mutagenicity. Moreover, the protective effect of each eggplant extract against oxidative DNA damage in human lymphocytes was investigated using the single-cell gel electrophoresis (comet) assay. The treatment cells with six eggplant extracts prevented DNA human lymphocytes in response to hydrogen peroxide, especially ML extract exhibited higher an inhibition percentage than other samples. This study demonstrated that these eggplants seem to be safe for consumption and their extracts could protect against DNA damage. Thus, these eggplants have the potential to provide health benefits associated with prevention or reduced risk of developing chronic diseases, such as cancer.