In vitro cytogenetic analysis of two different anti-phosphates (sevelamer hydrochloride and calcium carbonate) agents used by patients with hyperphosphatemia.

Sevelamer hydrochloride (SH) and calcium carbonate (CaCO3) are two agents included in the phosphate-binding group which are frequently prescribed in the treatment of patients with hyperphosphatemia. However, there are no satisfactory studies on the genotoxic effects of SH in vitro. This study was conducted to reveal the genotoxic and/or cytotoxic potential of these two drugs in cultured human peripheral lymphocytes. Human peripheral lymphocytes were treated with SH and CaCO3 at sublethal concentrations for 24 or 48 h for micronucleus assay and 1 h in the comet assay. CaCO3 and SH stimulated a slight increase in micronucleus formation however this increase was not significant compared to the control group. According to the findings of the comet test, only one concentration of the SH caused significant DNA damage (2 mg/ml, 48 h) whereas CaCO3 did not cause important DNA breakage. No significant oxidative damage or anti-radical effect caused by test substances was observed on the pure pBR322 plasmid DNA in a cell-free medium. Also, it was found that the drugs were devoid of mutagenic activity in the Ames test, but had a weak cytotoxic effect. Both test substances, particularly SH, significantly reduced the nuclear division index compared to the control group. In conclusion, the cytotoxic effect of SH was evident on the basis of in vitro tests and slightly higher than CaCO3.

The effects of titanium nanoparticles on enzymatic and non-enzymatic biomarkers in female Wistar rats.

Titanium dioxide (TiO2) nanoparticles (NPs) are widely used in industry, pharmacy, medicine, and food sectors. Therefore, this study deals with the effects of TiO2 NPs in female rats following oral administration in differing doses for 14 days (0, 0.5, 5, and 50 mg/kg b.w./d). The response of enzymatic biomarkers (Na,K-ATPase, Mg-ATPase, and AChE) was measured in the brain, kidney, and small intestine, while non-enzymatic biomarker levels, such as different forms of glutathione (GSH) and thiobarbituric acid reactive substances (TBARSs) were measured in the liver. The images of the tissues were obtained using a transmission electron microscope (TEM) to demonstrate TiO2 NP accumulation. Data showed that brain AChE activity decreased at all TiO2 NP doses, though brain ATPase activities increased. However, ATPase activities in the intestine and kidney did not change significantly. Levels of GSH forms did not change significantly, though there was a significant decrease in TBARS level at the highest NP dose. TEM images demonstrated that TiO2 NPs accumulated in a dose-dependent manner in the tissues. Data emphasized that the brain was the most sensitive organ against the effects of TiO2 NPs. This study suggests the need for further studies to evaluate better the toxic effects of TiO2 NPs.

Investigation of the genotoxic effects of patent blue V (E131) in human peripheral lymphocytes and in silico molecular docking.

Patent Blue V (PBV) is a water-soluble synthetic dyestuff that is used as a coloring agent in the food industry and for medical imaging in health monitoring. The aim of this study was to investigate the in vitro clastogenic, aneugenic and cytotoxic effects of PBV in human peripheral lymphocytes using micronucleus assay, comet assay, as well as plasmid DNA interaction and bacterial AMES tests. In addition to in vitro tests, the affinity of PBV against DNA was determined by molecular docking analysis in silico. PBV produced significant MN formation only at high doses and longer treatment time, however, it did not significantly affect the nuclear division index (NDI). Furthermore, PBV was unable to cause DNA single-strand breaks and significant oxidative damage on the pBR322 plasmid DNA and it didn't reverse the harmful effects caused by the clastogenic treatment of UV + H2O2 on plasmid DNA. In the Ames test, no significant increase was detected in the number of revertant colonies of mutant strains, TA98 and TA100, following PBV treatment. No significant molecular interaction between B-DNA and PBV occured in molecular docking simulations. In conclusion, PBV had no significant genotoxic and cytotoxic effects in this study. However, considering that the information intensity related to the genotoxic effects of PBV in the literature is still insufficient, reports of further studies with different genotoxicity endpoints will be needed to elucidate the exact genotoxic feature.

In vitro cytogenotoxic and mutagenic effects of Commiphora myrrha essential oil.

Commiphora myrrha, located in the tropical zone, is a widely used tree for medicinal purposes in the Arabian Peninsula and a large part of Africa. In this research, cytogenotoxic effects of the commercially available Commiphora myrrha essential oil (myrrh) were studied using micronucleus (MN), comet, and total oxidant (TOS), and total antioxidant (TAS) assays on human peripheral lymphocytes under in vitro conditions. In addition, pure pBR322 plasmid DNA was used to investigate DNA damaging/protecting activity of the essential oil. Finally, a bacterial reversion (Ames) test was performed using Salmonella typhimurium mutant strains TA98 and TA100 to determine the potential effect of the agent in the induction of gene mutations. The high concentration of Commiphora myrrha (0.125 µL/mL) induced MN formation significantly compared to the untreated control in both treatment times (24 or 48 h). Only at the highest concentration, nuclear division index (NDI) values were found lower than the controls. In the Comet test performed on healthy lymphocytes, only the highest concentration of myrrh caused significant increases in the percentage of damaged cells and genetic damage index (GDI) values. Myrrh oil showed no significant mutagenic effect on mutant Salmonella strains. In addition, the substance did not directly damage plasmid DNA but also protected DNA against damaging factors such as H2O2 and UV. Finally, in the TAS and TOS assays, no significant differences on the oxidative stress parameters were found in cell culture compared to the control. The results of this study showed that myrrh oil exerts cytogenotoxic risk only at higher concentrations.

Responses of biomarkers belonging to different metabolic systems of rats following oral administration of aluminium nanoparticle.

Nanoparticle (NP) forms of aluminium oxide (Al2O3) are used in various fields such as engineering, pharmacy, medicine etc. Compounds containing aluminium oxide NPs may present toxic effects after certain thresholds. Thus, the present study was carried out to determine the effects of Al2O3 nanoparticles (Al-NPs) in rats. For this aim, different doses (0, 0.5, 5, 50 mg/kg b.w./day) of Al NP (˜40 nm) were orally administered to female rats (Rattus norvegicus var. albinus) for 14 days and the response of several biomarkers such as activities of ATPases (total ATPase, Na,K-ATPase, Mg-ATPase) and acetylcholinesterase (AChE), levels of different glutathione forms and thiobarbituric acid reactive substances (TBARS) were measured in different tissues. Additionally, tissue accumulation of Al-NPs was demonstrated by a transmission electron microscope (TEM). The images showed the presence of Al-NP aggregates in all the tissues at all doses. The sizes of NP aggregates were dependent on NP doses and it was a bit more loose in the brain than in the liver and kidney. AChE activity in the brain decreased significantly at all NP doses, whereas TBARS levels in the liver did not alter significantly at any NP dose. Although there was no significant change in ATPase activities in the intestine at any NP dose, there were significant decreases in the kidney and brain. There were some variations in the levels of total glutathione (tGSH), oxidized glutathione (GSSG) and reduced glutathione (rGSH), though these variations were not significant (P > 0.05). Likewise, the ratio of rGSH/GSSG also did not differ significantly among NP doses and control. The brain seems most affected organ following Al-NP administration. This study demonstrated that most biomarkers in the tissues of rats were affected by Al-NP, showing the signal of toxic effects and suggests further studies to understand better the effects of Al NPs, especially in their use for pharmacology.

In vitro cytogenotoxic evaluation of sertraline.

Sertraline (SRT) is an antidepressant agent used as a neuronal selective serotonin-reuptake inhibitor (SSRI). SRT blocks serotonin reuptake and increases serotonin stimulation of somatodendritic serotonin 1A receptor (5-HT1AR) and terminal autoreceptors in the brain. In the present study, the genotoxic potential of SRT was evaluated using cytokinesis-block micronucleus (CBMN) cytome assay in peripheral blood lymphocytes of healthy human subjects. DNA cleavage-protective effects of SRT were analyzed on plasmid pBR322. In addition, biochemical parameters of total oxidant status (TOS) and total antioxidant status (TAS) in blood plasma were measured to quantitate oxidative stress. Human peripheral blood lymphocytes were exposed to four different concentrations (1.25, 2.5, 3.75 and 5 µg/mL) of SRT for 24- or 48-h treatment periods. In this study, SRT was not found to induce MN formation either in 24- or 48-h treatment periods. In contrast, SRT concentration-dependently decreased the percentage of MN and MNBN (r=-0.979, p<0.01; r=-0.930, p<0.05, respectively) when it was present for the last 48 hr (48-h treatment) of the culture period. SRT neither demonstrated a cleavage activity on plasmid DNA nor conferred DNA protection against H2O2. The application of various concentrations of SRT significantly increased the TOS and oxidative stress index (OSI) in human peripheral blood lymphocytes for both the 24- and 48-h treatment periods. Morover, the increase in TOS was potent as the positive control MMC at both treatment times. However, SRT did not alter the TAS levels in either 24- or 48-h treatment periods when compared to control. In addition, exposing cells to SRT caused significant decreases in the nuclear division index at 1.25, 2.50 and 3.75 µg/mL in the 24-h and at the highest concentration (5 µg/mL) in the 48-h treatment periods. Our results suggest that SRT may have cytotoxic effect via oxidative stress on cultured human peripheral blood lymphocytes.

Assessment of in vitro genotoxic and cytotoxic effects of flurbiprofen on human cultured lymphocytes.

Flurbiprofen is non-steroidal anti-inflammatory drug which is commonly used for its analgesic, antipyretic, and anti-inflammatory effects. The purpose of the study was to explore the genotoxic and cytotoxic effects of flurbiprofen in human cultured lymphocytes by sister chromatid exchange, chromosome aberration, and cytokinesis-blocked micronucleus tests. 10, 20, 30, and 40 µg/mL concentrations of flurbiprofen (solvent is DMSO) were used to treatment of human cultured lymphocytes at two different treatment periods (24 and 48 h). Flurbiprofen had no significant genotoxic effect in any of these tests. But exposing to flurbiprofen for 24 and 48 h led to significant decrease on proliferation index, mitotic index, and nuclear division index (NDI). Also, all decreases were concentration-dependent (except NDI at 24 h treatment period). Consequently, the findings of this research showed that flurbiprofen had cytotoxic effects in human blood lymphocytes.

Deferasirox-induced cytogenetic responses.

Deferasirox (commercially formulated as Exjade(®)) is one of the effective iron chelators used in treatment of iron overload diseases. In this study the effect of this substance for chromosome aberration, sister chromatid exchange and mitotic index was studied by in vitro (by using human peripheral lymphocytes) and in vivo (by using rat) analysis. Deferasirox increased the sister chromatid exchange frequency in all tested concentrations and periods in vitro. Also, in the presence of metabolic activator, the substance led to a statistically significant increase in the sister chromatid exchange frequencies only at high concentration. While in in vitro analysis the substance significantly increased abnormal cell percentages in all concentrations, in in vivo study the substance increased chromosome aberrations only in two concentrations at 12h treatment. In the cultured lymphocytes, deferasirox showed cytotoxicity by significantly reducing proliferation index and mitotic index values. While in the presence of metabolic activation it did not affect the proliferation index frequency, it had a stimulant effect on the mitotic index frequency. Deferasirox reduced significantly the mitotic index value in the bone marrow cells especially in high concentration and short treatment period (12h).

Signs of deferasirox genotoxicity.

Iron overload is a major health problem for patients who have to have continuous blood transfusions. It brings some metabolic problems together. Various iron chelating agents are being used for treatment of hemochromatosis which arises from excess iron accumulation. This study was conducted with the aim of determining whether deferasirox used as an iron chelator in patients with hemochromatosis has genotoxic effects. Commercial form of deferasirox, Exjade was used as test material. Test material showed a general mutagen character in mutant strains of Salmonella typhimurium. Deferasirox has also led to an increase in mutagenity-related polymorphic band count in random amplification of polymorphic DNA test done with bone marrow cells of rats. Similarly, test material has increased micronucleus formation in cultured in vitro human peripheral lymphocytes particularly in 48 h period. Consistently with the abovementioned findings, deferasirox reduced nuclear division index (NDI) compared to controls and some part of these reductions are statistically significant. NDI reductions were found at positive control levels at high concentrations.

In vitro genotoxic perspective of Tamiflu.

The aim of this study was to investigate the genotoxic and/or cytotoxic effects of Tamiflu, commercial form of the oseltamivir antiviral and most frequently prescribed for the treatment of influenza infections, on cultured human peripheral lymphocytes by using sister chromatid exchange (SCE), chromosomal aberration (CA), and cytokinesis-blocked micronucleus (CBMN) assays. Cells were treated with 0.5, 1, 2 µg/mL oseltamivir, the Tamiflu capsule ingredient, for 24 or 48 h in the absence or presence of an exogenous metabolic activation system (S9 mix). The test chemical did not demonstrate any genotoxic effect dose-dependently but it showed a weak cytotoxicity on cells in this study. On the other hand, some concentrations of Tamiflu (2 µg/mL without S9 mix for 48 h and 1 µg/mL with S9 mix) induced SCE and also decreased significantly the proliferation index (PI) (48 h period) and the nuclear division index (NDI) (24 h period) (P < 0.05) in the absence of S9 mix. Considering the results, Tamiflu did not induce significant increases of CA or micronucleated cells in vitro in cultured peripheral blood lymphocytes under the treatment conditions used but weak SCE induction was observed. On the other hand, the weak cytotoxic effects observed disappeared in the cultures treated in presence of the S9 mix.

The genotoxic and antigenotoxic effects of tannic acid in human lymphocytes.

The genotoxicity of tannic acid (TA, tannin) were investigated using chromosome aberration (CA), sister chromatid exchange (SCE), and micronucleus (MN) test systems in human peripheral lymphocytes. Also, the antigenotoxicity of TA against known mutagen EMS was also examined. The lymphocytes were treated with 1.74 × 10(-5), 3.49 × 10(-5), and 6.98 × 10(-5) µM of TA for 24- and 48-hour treatment periods. For the antigenotoxicity of TA, the lymphocytes were treated with three different concentrations of TA and 2.71 µM of EMS. TA synergically induced the CA alone and with the mixture of EMS. However, TA did not induce the SCE alone, whereas TA and EMS as a mixture also synergically induced SCE. TA alone showed no clear effect on micronucleus formation, and it did not induce the MN when used with EMS as a mixture. In addition, TA showed a synergistic cytotoxic effect by decreasing the mitotic and nuclear division indices. The replication index was decreased at all concentrations for 48 hours of treatment time by TA and EMS as a mixture.

Genotoxic potential of cyfluthrin.

Cyfluthrin (CAS no. 68359-37-5), a synthetic fluorinated pyrethroid insecticide, is widely used in the home environment and in agriculture because of its high activity against a broad spectrum of insect pests and its low animal toxicity. There are no adequate data on genotoxic effects of cyfluthrin. The aim of this study was to analyze the potential genotoxic effects of cyfluthrin. The genotoxicity of cyfluthrin was evaluated, in vitro, by assessing the ability of the insecticide to induce gene mutation (evaluated using the Ames/microsome test), chromosomal aberrations (CA), sister chromatid exchange (SCE) and micronucleus (MN) formation in cultured human peripheral blood lymphocytes. Additionally, CAs and cytotoxicity induced by cyfluthrin were investigated in rat (Rattus norvegicus var. Albinos) bone-marrow cells to assess in vivo genotoxicity of cyfluthrin. The counts of reverse mutations in Salmonella typhimurium were not significantly increased (P>0.05). The frequency of CAs in human lymphocytes, treated with any concentration of cyfluthrin (500, 1000 or 2000 microg/ml) for a 24-h period, was not significantly increased (P>0.05). In contrast, CA was significantly increased for the highest two concentrations (1000 and 2000 microg/ml) in the 48-h treatment group compared with the control group (dimethyl sulfoxide, DMSO). Micronucleus formation was significantly (P<0.05) increased for all doses after the 48-h treatment, although the frequency of SCE did not increase significantly (P>0.05). Mitotic index (MI), proliferation index (PI) and nuclear division index (NDI) decreased significantly (P<0.05) due to the potential cytotoxicity of cyfluthrin, especially after the 48-h treatment period. The frequency of chromosome aberrations in bone-marrow cells of rats treated with the test substance increased significantly (P<0.05) for all doses (250, 500 and 1000 mg/kg body weight) for the two treatment periods (12 and 24 h) and the two administration routes, viz. intraperitoneal injection (i.p.) and oral gavage (gvg). In vivo cytotoxicity of cyfluthrin was detected only after administration by gavage for the 24-h treatment period. All these findings were not dose-dependent.

The genotoxic effect of potassium metabisulfite using chromosome aberration, sister chromatid exchange, micronucleus tests in human lymphocytes and chromosome aberration test in bone marrow cells of rats.

Potassium metabisulfite (PMB) is used as an antimicrobial substance in many kinds of foods. In the present study, the effects of PMB on chromosome aberrations (CAs), sister chromatid exchanges (SCEs), and micronucleus (MN) formation in human lymphocytes and as well as its effect on CAs in bone marrow cells of rats were investigated. The human lymphocytes were treated with 25, 50, 100, and 200 microg/ml of PMB for 24 and 48 hr. PMB was also intraperitoneally (ip) injected to the rats as a single dose of 150, 300, and 600 mg/kg body weight (b.w.) for 12 and 24 hr before sacrifice. PMB induced abnormalities such as structural and numerical (total) CAs, SCEs, and MN formations in a dose dependent manner in the lymphocytes of the 24- and 48-hr treatment periods. In addition, PMB showed a cytotoxic effect by decreasing the replication index (RI), mitotic index (MI) and nuclear division index (NDI) in a dose dependent manner in human lymphocytes. The compound induced CA as well and decreased the MI in bone marrow cells of rats. It might be concluded that PMB had a high genotoxic and cytotoxic risk.

The effects of food protector biphenyl on sister chromatid exchange, chromosome aberrations, and micronucleus in human lymphocytes.

The aim of this study was to determine the possible genotoxic effects of biphenyl (E230), which is used as an antimicrobial agent in food by using sister chromatid exchanges (SCEs), chromosome aberrations (CAs), and micronucleus (MN) tests in human peripheral lymphocytes. The human peripheral lymphocytes were treated with four concentrations of biphenyl (10, 30, 50, and 70 microg/mL) for 24- and 48-h treatment periods. In the present study, biphenyl significantly increased the frequency of SCEs, CAs, and the frequency of MN when compared with both untreated control and solvent (dimethyl sulfoxide) control. The inductions of these abnormalities were in a dose-dependent manner. Biphenyl was capable to induce the structural CAs instead of numerical CAs. Biphenyl also showed a cytotoxic effect by decreasing the replication index at the highest two concentrations for 48 h and nuclear division index at the highest two concentrations for the 24- and 48-h treatment periods. However, biphenyl did not affect the mitotic index (MI).

The genotoxic effect of the new acaricide etoxazole.

Etoxazole is a member of the diphenyl oxazoline class of insecticide was newly developed for use on pome fruits, cotton and strawberries as a acaricide. In the present study, genotoxic effects of acaricide etoxazole (ETX) (miticide/ovicide) were investigated using chromosome aberration (CA) test, sister chromatid exchange (SCE) test and micronucleus test in human lymphocytes. ETX induced the CAs at all concentrations (5, 10 and 20 microg/ml) for 24 h and also induced the CA at the highest concentration (20 microg/ml) for 48 h only. The inducing the CAs for 48 hours treatment period was dose-dependent. Besides, it induced the SCE at all concentrations and treatment periods in a dose-dependent manner as well. Although, ETX decreased the mitotic index (MI) at all concentration and treatment periods dose-dependently, while it did not decrease the replication index (RI) when compared to the negative and solvent controls. In addition, ETX induced the micronucleus at all concentrations except 5 microg/ml for 48 h. This inducing was in a dose-dependent manner as well. In conclusion, it can be concluded that ETX has a potential genotoxic effects in cultured human peripheral lymphocytes.

Genotoxicity of aspartame.

In the present study, the genotoxic effects of the low-calorie sweetener aspartame (ASP), which is a dipeptide derivative, was investigated using chromosome aberration (CA) test, sister chromatid exchange (SCE) test, micronucleus test in human lymphocytes and also Ames/Salmonella/ microsome test. ASP induced CAs at all concentrations (500, 1000 and 2000 microg/ml) and treatment periods (24 and 48 h) dose-dependently, while it did not induce SCEs. On the other hand, ASP decreased the replication index (RI) only at the highest concentration for 48 h treatment period. However, ASP decreased the mitotic index (MI) at all concentrations and treatment periods dose-dependently. In addition, ASP induced micronuclei at the highest concentrations only. This induction was also dose-dependent for 48 hours treatment period. ASP was not mutagenic for Salmonella typhimurium TA98 and TA100 strains in the absence and presence of S9 mix.

Chromosome aberration and sister chromatid exchange in workers of the iron and steel factory of Iskenderun, Turkey.

The aim of this study was to investigate, by using chromosome aberration (CA) and sister chromatid exchange (SCE) tests, whether or not the workers employed in the Iskenderun (Turkey) iron and steel factory have any genotoxic risk. The CA and the SCE were investigated in 48 males employed in a coke ovens unit and 8 males employed in a product side unit of the factory and in control groups. The frequency of CA was higher while the frequency of the SCE was not in all the smoker-nonsmoker workers than in smoker-nonsmoker control groups. In addition, there was no significant decrease in the RI, while the MI was significantly lower than in the controls. .

No significant increase in chromosome aberrations and sister chromatid exchanges in cultured human lymphocytes treated with spiramycin.

In this study, the chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) were investigated in human lymphocytes treated with spiramycin antibiotic (trade name, rovamycin). Spiramycin did not induce the CAs and SCEs, and also did not decrease the mitotic index (MI). However, spiramycin decreased the replication index (RI) only at 48 h treatment times.