Cytokinesis-block Proliferation Index Research Articles

Human plasma can modulate micronucleus frequency in TK6 and OE33 cells in vitro

In this paper, we studied the potential genotoxic effects of human plasma from healthy volunteers, as well as patients with gastro-oesophageal reflux disease, Barrett’s oesophagus (BO) and oesophageal adenocarcinoma (OAC) using the oesophageal adenocarcinoma cell line (OE33) and the lymphoblastoid cell line (TK6). Both TK6 and OE33 cells were treated with plasma (10 % volume, replacing foetal bovine serum (FBS) or horse serum (HS)) at different time points of 4 h (for the micronucleus (Mn) assay and the invasion assay) and 24 h (for the cell cycle studies). Plasma-induced effects on DNA damage levels, cell viability and the cell cycle were studied by the micronucleus assay, cytokinesis block proliferation index (CBPI) and flow cytometry respectively. The expression of IL-8 in supernatants of TK6 cells and IFN-β in OE33 cells was also analysed by enzyme-linked immunosorbent assay (ELISA). Finally, we carried out an assessment of cellular invasion of OE33 cells following plasma treatment.The results of the micronucleus assay confirmed the genotoxicity of direct plasma treatment from some participants through the increase in DNA damage in TK6 cells. Conversely, some individual patient plasma samples reduced background levels of TK6 cell Mn frequency, in an anti-genotoxic fashion. In TK6 cells, (on average) plasma samples from patients with Barrett’s oesophagus induced higher micronucleus levels than healthy volunteers (p= 0.0019). There was little difference in Mn induction when using plasma versus serum to treat the cells in vitro. Cell cycle results showed that direct plasma treatment had a marked impact on OE33 cells at 24 h (p=0.0182 for BO and p=0.0320 for OAC) by decreasing the proportion of cells in the S phase, while plasma exposure was less impactful on the cell cycle of TK6 cells. Invasion of OE33 cells was also seen to be non-significantly affected by plasma treatment of OE33 cells.The addition of N-acetyl cysteine NAC in a dose-dependent matter did not alter the formation of Mn in TK6 cells, suggesting that reactive oxygen species (ROS) are not the root cause of plasma’s genotoxicity. The concentration of IL-8 in TK6 cells and IFN-β in OE33 cells was significantly higher in cells treated with OAC-derived plasma than in the untreated negative control. Collectively, our results demonstrate that plasma-specific effects are detectable which helps us better understand some important aspects of the biology of blood-based biomarkers under development.

Risk Characterization of Botanical Extracts Containing Hydroxyanthracenes as Determined by a Validated Micronucleus In Vitro Assay

Extracts of Rheum palmatum L., Rhamnus purshiana DC., Rhamnus frangula L., and Cassia senna L. are used in traditional medicine thanks to their beneficial properties. These species contain hydroxyanthracene derivatives, considered genotoxic and possibly related to colorectal cancer development. This research aimed to study, using a micronucleus assay in vitro, the genotoxic potential of Rheum palmatum L., Rhamnus purshiana DC., Rhamnus frangula L. (bark), and Cassia senna L. (leaves and fruits) extracts. The extracts were evaluated at different concentrations: from 0 to 2000 µg/mL for Rhamnus purshiana DC, from 0 to 2500 µg/mL for Rheum palmatum L. and Rhamnus frangula L., and from 0 to 5000 µg/mL for Cassia senna L. The cytokinesis-block proliferation index was calculated to analyse if the used concentrations showed cytotoxicity. The hydroxyanthracene content varied between 0.06% and 0.23% for aloe-emodin, and between 0.07% and 0.16% for emodin and rhein. No cytotoxic effect was detected at any of these concentrations. Micronucleus analyses showed a lack of genotoxicity for all the extracts tested. These results show that Rheum palmatum L., Rhamnus purshiana DC, Rhamnus frangula L., and Cassia senna L. extracts do not induce genotoxicity since no increase in micronuclei formation in human lymphocytes in vitro was detected.

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.

Cytogenotoxic evaluations of leaves and stems extracts of Rubus rosifolius in primary metabolically noncompetent cells

ABSTRACT Plants with medicinal potential may also produce adverse effects in humans. This seems to be the case for the species Rubus rosifolius, where preliminary studies demonstrated genotoxic effects attributed to extracts obtained from leaves and stems of this plant using on HepG2/C3A human hepatoma cells as a model. Considering the beneficial properties of this plant as an antidiarrheal, analgesic, antimicrobial, and antihypertensive and its effects in the treatment of gastrointestinal diseases, the present study was developed with the aim of determining the cytotoxic and genotoxic potential of extracts of leaves and stems of R. rosifolius in primary without metabolic competence in human peripheral blood mononuclear cells (PBMC). Cell viability analyses at concentrations of between 0.01 and 100 µg/ml of both extracts did not markedly affect cell viability. In contrast, assessment of the genotoxic potential using the comet assay demonstrated significant damage to DNA within PBMC from a concentration of 10 µg/ml in the stem extract, and a clastogenic/aneugenic response without cytokinesis-block proliferation index (CBPI) alterations at concentrations of 10, 20, or 100 µg/ml for both extracts. Under our experimental conditions, the data obtained demonstrated genotoxic and mutagenic effects attributed to extracts from leaves and stems of R. rosifolius in cells in the absence of hepatic metabolism.

Sulfoxaflor insecticide exhibits cytotoxic or genotoxic and apoptotic potential via oxidative stress-associated DNA damage in human blood lymphocytes cell cultures

The need for foodstuff that emerged with the rapidly increasing world population made fertilizers and pesticides inevitable to obtain maximum efficiency from existing agricultural areas. Sulfoxaflor is currently the only member of the new sulfoximine insecticide subclass of nicotinic acetylcholine receptor agonists. In the study, it was aimed to determine the in vitro genetic, oxidative damage potential, genotoxic and apoptotic effects of three different concentrations (10 µg/mL, 20 µg/mL and 40 µg/mL) of sulfoxaflor insecticide in the cultures of blood lymphocytes. In this study, the single-cell gel electrophoresis (comet), Cytokinesis Block Micronuclues Test (MN test), flow cytometry and measurement of Catalase (CAT) enzyme activity were used to determine genotoxic, apoptotic effects and oxidative damage potential, respectively. It found that there is a decrease in CPBI values and Live cell numbers. It was observed an increase in late apoptotic and necrotic cell numbers, Micronucleus frequency, and Comet analysis parameters (GDI and DCP). There is a significant difference between negative control and all concentration of insecticide for Cytokinesis Block Proliferation Index (CBPI) values and late apoptotic, necrotic and viable cell counts. An increase in CAT enzyme levels was observed at 10 and 20 µg/mL concentrations compared to control., It is found that CAT enzyme activity was inhibited at concentrations of 40 µg/mL. This study is crucial as it is the first study to investigate the impact of Sulfoxaflor insecticide on peripheral blood lymphocyte cells. The genotoxic, oxidative damage, and apoptotic effects of Sulfoxafluor insecticide on the results obtained and its adverse effects on other organisms raise concerns about health and safety.

Genotoxic potential of different nano-silver halides in cultured human lymphocyte cells

Most antibacterial applications in nanotechnology are carried out using silver nanoparticles (AgNPs). However, there is a dearth of information on the biological effects of AgNPs on human blood cells. In this study, the cytotoxic and genotoxic potentials of ionic silver (Ag+), AgNP, silver bromide (AgBr), silver chloride (AgCl), and silver iodide (AgI) were evaluated through chromosome aberration (CA) test and cytokinesis-blocked micronucleus (CBMN) test in human cultured lymphocytes in vitro. Furthermore, the potential damages that can cause to DNA were evaluated through alkaline single cell gel electrophoresis (Comet) assay on isolated lymphocytes. The results showed that AgNPs exerted cytotoxic effects by reducing the cytokinesis-block proliferation index and mitotic index at 24 and 48 h. AgNPs also increased micronucleus (MN) formation at both exposure times in the cultured cells. Meanwhile, AgCl had no genotoxic effects on the human lymphocyte cultured cells but had a cytotoxic effect at high doses. AgNP, Ag+, AgBr, and AgI caused substantial DNA damage by forming DNA strand breaks. They may also have clastogenic, genotoxic and cytotoxic effects on human lymphocyte cells. Based on the foregoing findings, silver nanomaterials may have genotoxic and cytotoxic potentials on human peripheral lymphocytes in vitro.

Modulatory effect of myricitrin against chromosome instability and cytostasis induced by bleomycin and oxaliplatin in CHO-K1 cells

Myricitrin (MYR), a flavonol consumed in the leaves and fruits of plants of the Myrtaceae family, presents anti-proliferative, anti-inflammatory, anti-diabetic, and antioxidant properties in humans. However, there are few studies regarding the cyto-genotoxicity and the chemopreventive potential of MYR. Using the in vitro Micronucleus test, the cytostasis, mutagenicity, and modulatory effect of MYR in CHO-K1 cells were assessed. The concentrations of 39 and 78 µg/mL (p < 0.001.) of MYR decrease the cytokinesis-block proliferation index (CBPI) in the short exposure treatment (4 h), while in the extended treatment (24 h), concentrations of 4.8, 9.7, 19.5, 39 and 78 µg/mL (p < 0.001.) decreased the CBPI. MYR associated with oxaliplatin decreased CBPI at all tested concentrations in the pre-(p < 0.001) and post-treatments (p < 0.001), but there was no decrease when associated with bleomycin. As for chromosome instability, MYR did not increase the frequency of micronuclei (MNi), nucleoplasmic bridges (NPBs), or nuclear buds (NBUDs) in the 4 h exposure time, however, in the 24 h treatment, MYR increased the frequency of MNi and NPBs at concentration 19.5 µg/mL (p < 0.001). As for the modulatory effect, MYR associated with bleomycin decreased the frequency of MNi, NPBs, and NBUDs at all concentrations in the pretreatment (MNi and NPBs p < 0.001, NBUDs p < 0.05) and simultaneously (MNi, NPBs and NBUDs p < 0.001). When associated with oxaliplatin, the simultaneous treatment decreased the frequency of MNi (p < 0.001) and NBUDs (p < 0.01) at all concentrations, however, in the post-treatment, MYR increased MNi (p < 0.001) and NPBs p < 0.05) in CHO-K1 cells, when compared to oxaliplatin alone. The results demonstrated that MYR could modulate the mutagenic and cytostatic actions of bleomycin and oxaliplatin, demonstrating distinct behaviors, depending on the mechanism of action of the chemotherapeutic agent.

Genetic instability of lung induced by carbon black nanoparticles is related with Plk1 signals changes

As a possible carcinogen, carbon black has threatened public health. However, the evidences are insufficient and the mechanism of carcinogenesis is still not specified. Thirty rats were randomly divided into 3 groups, namely 0, 5 and 30 mg/m3 Carbon Black nanoparticles (CBNPs) groups, respectively. Rats were treated with CBNPs by nose-only inhalation for 28 days, 6 h/day. The human bronchial epithelial (16HBE) cells were treated with 0, 50, 100 and 200 μg/mL CBNPs for 24 h. Polo-like kinase 1 (PLK1) overexpression cell line was established by pcDNA3.1-PLK1 stable transfection. Our results showed that CBNPs exposure could induce DNA damage and genetic changes as well as apoptosis in vivo and in vitro. The DNA repair ability increased after CBNPs exposure. Cell cycle process was retarded at the G2/M phases in 16HBE cells after CBNPs treatment. The PLK1, ChK2 GADD45α and XRCC1 expression levels changed in rat lung and 16HBE cells after CBNPs treatment. Compared with NC 16HBE cells, DNA damage and repair, numbers of apoptotic cells and micronucleus (MN) rates, as well as the ChK2, GADD45α, XRCC1 expression levels decreased, whereas cytokinesis block proliferation index (CBPI) and replicative index (RI) increase in PLK overexpression (PLK+/+) cells after CBNPs treatment. This study highlighted that PLK1 related with the genetic toxicity of CBNPs in vitro and in vivo. Our results provided evidences supporting reclassification of carbon black as a human possible carcinogen.

The Alleviating Effect of Herniarin Against Ionizing Radiation-Induced Genotoxicity and Cytotoxicity in Human Peripheral Blood Lymphocytes.

Herniarin is a simple coumarin that is found naturally in some plant species. In this study, we aimed to evaluate the protective effect of herniarin against ionizing radiation-induced genotoxicity and cytotoxicity in human peripheral blood lymphocytes. Herniarin was added to human lymphocytes before irradiation with a dose of 2 Gy of Xrays. The antagonistic potential of herniarin against radiation was measured by MTT [3-(4,5- dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] assay, micronucleus assay, flow cytometry, and reactive oxygen species (ROS) level analysis. The maximum survival of lymphocytes against radiation was observed at a concentration of 50 μM of herniarin and a treatment time of 1 h. Pretreatment with 50 μM herniarin significantly decreased the micronuclei frequency, the percentage of apoptotic lymphocytes, and the ROS level in irradiated human lymphocytes. Moreover, 50 μM herniarin significantly increased the cytokinesis blocked proliferation index in irradiated lymphocytes. Herniarin could reduce radiation-induced cytotoxicity and genotoxicity in human lymphocytes. To complete the results of this study, it is suggested that in the future, more preclinical studies with larger samples or animal models be performed on herniarin.

Assessment of genetic damage induced by gadolinium-based radiocontrast agents

BackgroundToday, although gadolinium based contrast agents have been frequently used in the field of medicine, there is limited data available whether gadolinium based agents affect the genome. AimThe present study aimed to investigate the genotoxic and cytotoxic potentials of gadoteric acid and gadoversetamide used as gadolinium-based contrast agents for magnetic resonance (MR) imaging. Material and methodsThe cytokinesis-block micronucleus assay was applied to human peripheral blood lymphocytes to assess the genotoxicity measured as micronucleus (MN), nucleoplasmic bridge (NPBs) and nuclear bud (NBUDs) frequencies. Furthermore, cytokinesis-block proliferation index (CBPI) was calculated to determine cytostasis. Lymphocytes were treated with gadoteric acid at concentrations of 1.0, 2.5, 5.0, and 25 mM and with gadoversetamide at concentrations of 0.25, 1.0, 2.5, and 5.0 mM for 48 h. ResultsGadoteric acid did not cause significant increase in MN, NBPs and NBUDs frequencies and CBPI values at any concentration. Gadoversetamide induced significantly increase MN formation at concentration of 2.5 mM, NBP formation at concentrations of 1.0 and 2.5 mM, and NBUD formation at concentrations of 0.25, 1.0 and 2.5 mM. Additionally, gadoversetamide exposure resulted in statistically significant decrease in CBPI values compared to the control at concentrations of 2.5 and 5.0 mM. In addition, CBPI levels in response to concentrations of gadoversetamide was negatively and significantly associated with concentration. ConclusionThese findings show that gadoteric acid does not have genotoxic or cytotoxic potential, while gadoversetamide might have both genotoxic and cytotoxic potential on human peripheral blood lymphocytes. As a comparison, gadoversetamide was found more genotoxic and cytotoxic.

Assessment of the Biological Effects of Pellia endiviifolia and its Constituents in Vitro

Liverworts are characterized by a high content of bioactive compounds reported to show antimicrobial, anticancer, and antioxidant properties. In this study, the biological effects of the methanol extract of the liverwort Pellia endiviifolia and its constituents, bis-bibenzyls perrottetin E, 10′-hydroxyperrottetin E, and 10,10′-dihydroxyperrottetin E, were investigated using human peripheral blood cells as a model system. The assessment of the investigated compounds comprised testing their genotoxicity, apoptotic potential, and redox modulating activities. The genotoxicity testing indicated that medium (25 µM) and high concentrations (100 µM) of the investigated compounds displayed genotoxic and antiproliferative effects in human lymphocytes as revealed by significant, concentration-dependent enhancement of the micronuclei incidence and decrease in the cytokinesis-block proliferation index compared to the control ( P &lt; .001). Analysis of leukocyte apoptosis showed a substantial potential of all investigated compounds to induce apoptosis, which was not concentration-dependent. The P endiviifolia extract and perrottetin E demonstrated considerable pro-apoptotic potential, even at the lowest concentration (1 µM) applied. Evaluation of the redox modulating effects, which comprised measuring erythrocyte catalase activity and the lymphocyte malondialdehyde level, showed that the investigated compounds did not induce oxidative stress in human peripheral blood cells ( P &gt; .05). The observed genotoxic, antiproliferative, and proapoptotic effects of the investigated compounds make them suitable for further comprehensive studies related to their possible applications as anticancer agents.

Assessment of the genotoxic potential of tetrachlorvinphos insecticide by cytokinesis-block micronucleus and sister chromatid exchange assays.

Tetrachlorvinphos is an organophosphate that is classified as a carcinogen in humans by several authorities. Due to very limited data regarding the genotoxic potential, we aimed to comprehensively investigate in vitro genotoxic potential of tetrachlorvinphos. We performed our study by applying the cytokinesis-block micronucleus cytome and sister chromatid exchange (SCE) assays to human peripheral blood lymphocytes. We evaluated micronucleus (MN) and SCE frequencies and cytokinesis-block proliferation index in both exposed and non-exposed lymphocytes. We also calculated the chromosomal instability level in response to exposure by combining the results of MN and SCE. We found that MN frequency did not increase with exposure to tetrachlorvinphos (0-50µg/ml). In contrast, we observed that SCE frequencies significantly increased with exposure to ≥5µg/ml tetrachlorvinphos. Furthermore, exposure to tetrachlorvinphos at concentrations of 50µg/ml induced a significant increase in chromosomal instability level (p < 0.05). Cytokinesis-block proliferation index level did not significantly decrease in response to tetrachlorvinphos exposure. Our findings reveal that tetrachlorvinphos resulted in different DNA damages that were measured by two assays. Furthermore, our findings suggested that exposure to tetrachlorvinphos increased chromosomal instability that is a hallmark of many malignancies. We conclude that although tetrachlorvinphos does not significantly increase the MN level, the significant increase of both SCE and CIN frequencies indicates the genotoxic potential of tetrachlorvinphos in human peripheral lymphocytes. Additionally, tetrachlorvinphos is not cytotoxic in the range of tested concentrations.

Genotoxic and cytotoxic effects of polyethylene microplastics on human peripheral blood lymphocytes

Currently, we need emerging initial data regarding how plastic exposures affect cellular and molecular components and how such interactions will be crucial for human health. We aimed to determine the genotoxic and cytotoxic effects of microplastic (MPs,10-45 μm, polyethylene) on human peripheral lymphocytes by using the cytokinesis-block micronucleus cytome (CBMN) assay, which is a comprehensive method to reveal a range of mechanisms, not only diseases but also response to environmental exposures. We measured micronucleation (MN), nucleoplasmic bridge formation (NPB), and nuclear bud formation (NBUD) in human peripheral blood lymphocytes. We also measured the cytokinesis-block proliferation index (CBPI) to calculate cytostasis, which indicates cytotoxicity in lymphocytes treated with five different MPs concentrations for 48 h. Even lower concentrations of MPs increased the level of genomic instability. We found that the in vitro MP exposure significantly increased MN, NPB, and NBUD frequencies. Since we investigated the effect of larger particles relative to the lymphocytes, mechanic interaction of MPs with cells, the release of monomer and additives from MPs could be suggested as possible mechanisms accounting for increasing genomic instabilities. We did not observe a decrease in the cell proliferation index, indicating a lack of MPs’ cytotoxic potential. To the best of our knowledge, our study is the first to identify MPs’ genotoxic potential in human peripheral blood lymphocytes. We suggested further studies to investigate the genotoxic and cytotoxic potential of smaller plastics and the chronic effect of MP on the human population.

Micronucleus Assay in Environmental Biomonitoring

Abstract Nowadays many chemicals are widely used in agriculture to ensure high crop yields or in veterinary/human medicine to cure diseases. After their improper usage they may contaminate the environment, persist in it and adversely affect both the target and/or the non-target organisms. One of the ways to detect the occurrence of chemicals in the environment is to assess their impact on aquatic and farm animals; both are directly or indirectly exposed via their feed and water. The micronucleus assay is a standardly used cytogenetic test for the simultaneous detection of clastogenic and aneugenic agents. Additionally, cytotoxic effects are also assessed by analysing the proliferation changes using the cytokinesis-blocked proliferation index. The occurrence of micronuclei is analysed in many types of cells like the peripheral blood cells, bone marrow or cell lines according to standards for micronuclei detection. The analysis of published results has shown that the micronucleus assay is, together with the chromosomal aberration test, one of the most often used test in genotoxicity assessment. Its results have contributed to reassessing the use of multiple chemicals available on the market. Moreover, it is a compulsory test before approving the chemical/ pesticide for the market.

Phospholipids modifications in human hepatoma cell lines (HepG2) exposed to silver and iron oxide nanoparticles

Metallic nanoparticles such as silver (Ag NPs) and iron oxide (Fe3O4 NPs) nanoparticles are high production volume materials due to their applications in various consumer products, and in nanomedicine. However, their inherent toxicities to human cells remain a challenge. The present study was aimed at combining lipidomics data with common phenotypically-based toxicological assays to gain better understanding into cellular response to Ag NPs and Fe3O4 NPs exposure. HepG2 cells were exposed to different concentrations (3.125, 6.25, 12.5, 25, 50 and 100µg/ml) of the nanoparticles for 24h, after which they were assayed for toxic effects using toxicological assays like cytotoxicity, mutagenicity, apoptosis and oxidative stress. The cell membrane phospholipid profile of the cells was also performed using shotgun tandem mass spectrometry. The results showed that nanoparticles exposure resulted in concentration-dependent cytotoxicity as well as reduced cytokinesis-block proliferation index (CBPI). Also, there was an increase in the production of ROS and superoxide anions in exposed cells compared to the negative control. The lipidomics data revealed that nanoparticles exposure caused a modulation of the phospholipidome of the cells. A total of 155 lipid species were identified, out of which the fold changes of 23 were significant. The high number of differentially changed phosphatidylcholine species could be an indication that inflammation is one of the major mechanisms of toxicity of the nanoparticles to the cells.

Assessment of the genotoxic potential of a migraine-specific drug by comet and cytokinesis-block micronucleus assays

ABSTRACTBackground: Eletriptan is a migraine-specific drug-containing the triptan group. In terms of drug safety, the present study aimed to investigate the genotoxic potential of eletriptan.Research design & methods: We conducted our study by using the cytokinesis-block micronucleus cytome (CBMN) assay, a comprehensive method for measuring micronucleus formation, and a sensitive method for detecting DNA-strand breaks. In the assay, cytokinesis-block proliferation index and the frequency of micronuclei were evaluated in lymphocytes treated with three different concentrations (1, 10 and 25 µg/ml) of eletriptan for 48 hours. In comet assays, DNA damage was evaluated in leucocytes treated with three different concentrations (1, 10 and 25 µg/ml) of eletriptan for an hour.Results: Eletriptan did not induce cytotoxicity nor any increased micronuclei frequencies. While the comet parameters % DNA in tail, tail moment, and the olive moment was found to be significantly increased at 10 and 25 µg/ml, the cytokinesis-block proliferation index values were not.Conclusion: These findings suggest that eletriptan is non-cytotoxic but potentially weakly genotoxic at higher concentrations (10 and 25 µg/ml).

Cytotoxic and genotoxic effects of Kalanchoe pinnata (Lam.) Pers. fresh leaf juice in the cultured human blood lymphocytes

This is the first investigation on the in vitro cytotoxicological and genotoxicological effects of Kalanchoe pinnata (Lam.) Pers. fresh leaf juice, for establishing a safe and effective quantity for use. Peripheral blood of 6 healthy, non-addicted males between 20 and 25 years of age was used for toxicity assessment by 3-(4, 5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), mitotic index (MI), sister chromatid exchanges (SCEs) and Cytokinesis Blocked Micronucleus (CBMN) assays. The IC50 of the leaf juice calculated by MTT assay was 155.1 µl. The 50 and70 µl of the juice concentrations were decided after standardization by MI and showed non-significant and significant decrease respectively when compared to control. The SCEs/Cell and SCEs/Chromosome were increased non-significantly (50 µl) and significantly (70 µl),while cell cycle proliferative index, Average generation time and Population doubling time values were non-significant for both doses when compared to the controls. In CBMN assay, the Cytokinesis block proliferation index, cytotoxicity, Micronuclei, Nuclear bud, Nucleoplasmic bridge frequencies and total DNA damage biomarkers showed non-significant changes for both 50 and 70 µl. The changes observed were significant only at 70 µl for MI and SCEs, which were significantly lower than that by positive control indicating a non genotoxic effect. Hence, the fresh leaf juice can be used pharmaceutically as well as traditionally, but for long durations and higher doses should be used with caution, as it can have mutagenic effect at particularly high levels.

Mutagenic and cytostatic activities of the Xylopia laevigata essential oil in human lymphocytes

Hydro-distilled essential oil from leaves of Xylopia laevigata was characterized by GC-MS. Twenty-seven components were identified and the oil’s major constituents comprised germacrene D, bicyclogermacrene, (E)-caryophyllene and germacrene B. The cytotoxicity of the essential oil of X. laevigata (EOXL), determined by MTT and mitotic index methods in cultured human lymphocytes was observed in all tested concentrations. Cultures treated with EOXL demonstrated significant increase in the frequencies of micronuclei in the cytokinesis-block micronucleus assay (CBMN) and reduction of the cytokinesis-block proliferation index (CBPI) rates. Results demonstrated the cytostatic and mutagenic effects of EOXL, the latter for the first time.

Contrast-induced nephropathy in an animal model: Evaluation of novel biomarkers in blood and tissue samples.

Identification of novel biomarkers of contrast-induced nephropathy (CIN) that may more accurately detect renal function changes; reflect kidney damage; assist monitoring; and elucidate pathophysiology attract considerable scientific attention nowadays. To evaluate novel biomarkers of nephrotoxicity in blood/tissue samples of a CIN model, 10 New Zealand white rabbits were divided into group 1 (n = 5; iopromide) and group 2 (n = 5; control). Blood was drawn at 0 h (immediately), 24 h and 48 h after contrast medium (CM) administration. Animals were euthanized at 48 h and kidneys were removed. Serum creatinine (sCr)/symmetric-asymmetric dimethylarginine (SDMA-ADMA) levels were measured. CM genotoxic/cytotoxic effect was investigated 48 h post-CM exposure using micronucleus assay in lymphocytes. Cytological examination was conducted using touch preparation technique (TPT). All animals in group 1 developed CIN: mean sCr levels increased by 68.2% within 48 h. Significant SDMA-ADMA level elevation was observed at 0 h and 24 h with insignificant drop at 48 h in group 1, remaining normal in group 2 at all time-points. Significant increase in bi-nucleated cells with micronuclei and micronuclei frequency was detected in group 1. Cytokinesis block proliferation index was reduced insignificantly in group 1. TPT revealed degenerative lesions/inflammation, cell degeneration, abnormal uterine tubular casts and rubella in kidneys of all animals in group 1. Group 2 presented normal cells.

Antineoplastic Activity and Genotoxicity of Organic Extracts and Barbatic Acid Isolated from the Lichen Cladonia salzmannii Nyl

Background: Secondary metabolites are responsible for most of the biological activities of lichens. Many of these compounds exhibit significant antineoplastic activity. The aim of the present in vitro study was to evaluate the cytotoxic and genotoxic activities of organic extracts and purified barbatic acid from the lichen Cladonia salzmannii (Nyl.).&#x0D; Methods: The thallus of the lichen (22 g) was cleaned and dried with the solvents diethyl ether, chloroform and acetone. Organic extracts were obtained using the hot exhausted method in a Soxhlet apparatus. Barbatic acid (BAR) was purified from the ether extract (1.3 g). Chemical analysis of the organic extracts and purified BAR was performed using thin-layer chromatography. The purity of purified BAR was determined using high-performance liquid chromatography. The MTT method [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and cytokinesis-block proliferation index (CBPI) were used to determine the cytotoxic activity of the organic extracts and purified BAR. The micronucleus test and comet assay were used to determine genotoxic potential of the organic extracts and purified BAR. Dimethyl sulfoxide was used as the diluting solvent of the samples in all biological tests.&#x0D; Results: The IC50 results demonstrated significant cytotoxic potential of the ether extract (50 µg/mL) against cell lines NCI-H292 (IC50: 29.91 µg/mL), HEp-2 (IC50: 26.75 µg/mL) and HL-60 (IC50: 3.59 µg/mL) as well as the purified BAR (25 µg/mL) against cell lines HEp-2 (IC50: 15.79 µg/mL) and MCF-7 (IC50: 18.28 µg/mL). The CBPI demonstrated the cytotoxic activity of the purified BAR at all concentrations tested (5, 10, 20 and 40 µg/mL) and all organic extracts (50 µg/mL) against Ehrlich carcinoma cells. For sarcoma 180, only BAR purified at a concentration of 40 µg/mL and the ether and chloroform extracts (50 µg/mL) were considered cytotoxic. The micronucleus test revealed that the purified BAR at a concentration of 5 µg/mL had no genotoxic potential against either tumor cell line. Furthermore, the chloroform extract and purified BAR at a concentration of 10 µg/mL were not considered genotoxic for sarcoma 180. In the comet assay, all compounds tested induced DNA damage in both tumor lines.&#x0D; Conclusion: Based on the present results, organic extracts and purified barbatic acid from C. salzmannii exhibit cytotoxic and genotoxic activity against of the tumor cell lines tested.