Molecular genetic and biochemical responses in human airway epithelial cell cultures exposed to titanium nanoparticles in vitro.

Titanium nanoparticles (NPs) have very wide application areas such as paint, cosmetics, pharmaceuticals, and biomedical applications. And, to translate these nanomaterials to the clinic and industrial domains, their safety needs to be verified, particularly in terms of genotoxicity and cytotoxicity. Therefore, in this study, we aimed to investigate of cytotoxicity and changes in gene expression profiles influenced by commonly titanium (as titanium carbide, titanium carbo-nitride, titanium (II) oxide, titanium (III) oxide, titanium (IV) oxide, titanium nitride, titanium silicon oxide) NPs in human alveolar epithelial (HPAEpiC) and pharynx (HPPC) cell lines in vitro since inhalation is an important pathway for exposure to these NPs. HPAEpiC and HPPC cells were treated with titanium (0-100 µg/mL), NPs for 24 and 48 h, and then cytotoxicity was detected by, [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide] (MTT), uptake of neutral red (NR) and lactate dehydrogenase (LDH) release assays, while genotoxicity was also analyzed by cDNA array - RT-PCR assay. According to the results of MTT, NR and LDH assays, all tested NPs induced cytotoxicity on both HPAEpiC and HPPC cells in a time- and dose-dependent manner. Determining and analyzing the gene expression profiles of HPAEpiC and HPPC cells, titanium NPs showed more changes in genes related to DNA damage or repair, oxidative stress, and apoptosis. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2056-2064, 2017.

Anticancer, Antioxidant and Cytotoxic Potential of Thymol in vitro Brain Tumor Cell Model.

BACKGROUND: Thymol (THY), which is a monocyclic monoterpene, found in oil of thyme various other kinds of plants. Until today, although different biological properties of THY have been indicated, its neurological toxicity has never been investigated. METHOD: In this study, in vitro antiproliferative (by 3-(4,5 dimetylthiazol-2-yl)-2,5 diphenlytetrazolium bromide (MTT) test), genotoxic (by single cell gel electrophoresis (SCGE)) and oxidative effects (by total antioxidant capacity (TAC) and total oxidative status (TOS) analysis) of THY (0-400 mg/L) were assessed on cultured primary rat neurons (CPRNs) and N2a neuroblastoma cells. RESULTS: The obtained data from MTT analysis revealed that THY (only at 400 mg/L) led to significant (p<0.05) decreases of the cell viability in cultured primary rat neurons. And, THY was found to inhibit cell growth in N2a cells at concentrations of 200 and 400 mg/L. Again, DNA damage rates were statistically indifferent (p>0.05) in both treated cell type as compared to control group. The present results also showed that 10, 25 and 50 mg/L of THY application into the cell cultures supported antioxidant capacity in primary rat neurons but not in N2a cells. CONCLUSION: In a conclusion, these results confirm that THY may have antiproliferative potential against brain tumor cells involving oxidative alteration.

Development of glycine-α-methyl-proline-containing tripeptides with neuroprotective properties.

Herein is described the synthesis of novel glycine-α-methyl-proline-containing tripeptides (GP(Me)X tripeptides namely GP(Me)R, GP(Me)K, and GP(Me)H) with the aim of obtaining derivatives highly stable in human plasma and able to counteract neuroinflammatory processes that are distinctive of neurodegenerative pathologies. The syntheses of GP(Me)R, GP(Me)K, and GP(Me)H were all achieved both by introducing the Pro(Me) residue into the Gly-Pro-Arg (GPR) sequence in place of the native Pro in P2 position and replacing the basic amino acid Arg in P3 position by Lys or His. Results showed that all novel GP(Me)X tripeptides are stable in human plasma (t1/2 > 51 h) and that GP(Me)H - generating stable intramolecular H-bond in a C11-turn by interaction of His imidazole ring and Gly carbonyl group - restored physiological levels of nitric oxide deriving from neuronal NOS (nNOS) activity, thus preventing the inflammatory response by suppression of the NF-kB activity and, consequently, the expression of inflammatory genes such as inducibile NOS (iNOS). Therefore, GP(Me)H could be a lead compound for further development of peptidomimetics able to contrast neuroinflammatory processes.

In vitro assessment of cytogenetic and oxidative effects of α-pinene.

α-Pinene (α-pinene), a bicyclic monoterpene, is present in the oils of many species of coniferous trees, most notably the pine, and is known for its diverse biological properties such as antimicrobial, anti-inflammatory, antiproliferative and antioxidant. However, there are limited data on the cytogenetic and antioxidant effects of α-pinene in cultured human blood cells (n = 5) for the first time. The purpose of this study was to investigate the genetic, oxidative, and cytotoxic effects of α-pinene in cultured human blood cells (n = 5) for the first time. Human blood cells were treated with α-pinene (0 to 200 mg/L) for 24 and 48 h, and then cytotoxicity was detected by lactate dehydrogenase (LDH) release and (3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide) (MTT) assay, while DNA damage was also analyzed by micronucleus (MN) assay, chromosomal aberration (CA) assay and 8-oxo-2-deoxyguanosine (8-OH-dG). In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative stress (TOS)) were examined to determine oxidative effects. The results of LDH and MTT assays showed that α-pinene (at 200 mg/L) decreased cell viability. In our in vitro test systems, it was observed that α-pinene did not cause any statistically important changes in the rates of studied genotoxicity endpoints but dose-dependent alterations were observed in TAC and TOS levels. α-Pinene treatment caused increases in TAC levels (at 25 and 50 mg/L) and decreases in TOS levels (only at 200 mg/L) on human lymphocytes. In conclusion, the findings of the present study confirm for the first time that α-pinene could be a significant source of natural antioxidant compound that may have beneficial health effects.

In vitro risk assessment of usnic acid.

Lichens are symbiotic organisms composed of fungi and algae and are very common in Turkey. Lichen secondary metabolites are mainly phenolic compounds produced by fungal partner of lichen symbiosis. Usnic acid (UA) is one of the most common lichen metabolites, and it was reported that to be effective for a wide range of pharmacological purposes including antiviral, antitumor, and antiprotozoal. However, there are limited data on the genotoxic and antioxidant effects of UA in cultured human peripheral blood cells. Therefore, the aim of this thesis study was to investigate the genetic and oxidative effects of UA in cultured human blood cells (n = 5). The UA was added into culture tubes at various concentrations (0-200 µg/ml). Chromosomal aberrations (CA) and micronuclei (MN) tests were performed for genotoxic damage influences estimation. In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative status (TOS)) were examined to determine oxidative effects. In our in vitro test systems, it was observed that UA had no mutagenic effects on human lymphocytes. Furthermore, our results indicated that low concentrations (1 and 5 µg/ml) of UA caused increases of TAC levels in cultured human blood cells. And, the TOS levels were not changed (p > 0.05) when all the concentrations (except for 200 µg/ml) of UA were applied. In conclusion, UA can be a new resource of therapeutics as recognized in this study with their nonmutagenic and antioxidant features.

Investigation of cytotoxic, genotoxic and oxidative properties of carvacrol in human blood cells.

Carvacrol (CVC), a major constituent of genera Origanum and Thymus, is such a substance that has attracted attention because of its wide variety of beneficial biological activities such as antibacterial, antifungal, and anticancer effects. However, there are limited data on the cytogenetic and antioxidant effects of CVC in cultured human blood cells. The aim of this study was to investigate for the first time the genetic, oxidative, and cytotoxic effects of CVC in cultured human blood cells (n= 5). Human blood cells were treated with CVC (0-200 mg/L) for 24 and 48 h and then cytotoxicity detected by lactate dehydrogenase (LDH) release and (3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide) (MTT) assay, while DNA damage was also analyzed by micronucleus (MN) assay, chromosomal aberration (CA) assay and 8-oxo-2-deoxyguanosine (8-OH-dG) level. In addition, biochemical parameters (total antioxidant capacity [TAC] and total oxidative stress [TOS]) were examined to determine the oxidative effects. The results of LDH and MTT assays showed that CVC (at concentrations above 100 mg/L) decreased cell viability. In our in vitro test systems, it was observed that CVC had no mutagenic effects on human lymphocytes. On the other hand, CVC (at 50, 75, and 100 mg/L) treatment caused statistically important (p< 0.05) increases in TAC and TOS levels (at 150 and 200 mg/L) on human lymphocytes. In conclusion, CVC can be a new resource of therapeutics as recognized in this study with their nonmutagenic and antioxidant features.

Genotoxic and oxidative damage potentials in human lymphocytes after exposure to terpinolene in vitro.

Terpinolene (TPO) is a monocyclic monoterpene found in the essential oils of various fir and pine species. Recent reports indicated that several monoterpenes could exhibit antioxidant effects in both human and animal experimental models. However, so far, the nature and/or biological roles of TPO have not been elucidated in human models yet. The aim of this study was to investigate the genetic, oxidative and cytotoxic effects of TPO in cultured human blood cells (n = 5) for the first time. Human blood cells were treated with TPO (0-200 mg/L) for 24 and 48 h, and then cytotoxicity was detected by lactate dehydrogenase (LDH) release and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assay, while DNA damage was also analyzed by micronucleus assay, sister chromatid exchanges assay and 8-oxo-2-deoxyguanosine (8-OH-dG) level. In addition, biochemical parameters [total antioxidant capacity (TAC) and total oxidative stress (TOS)] were examined to determine oxidative effects. The results of LDH and MTT assays showed that TPO (at concentrations greater than 100 mg/L) decreased cell viability. In our in vitro test systems, it was observed that TPO had no genotoxicity on human lymphocytes. Again, TPO (at 10, 25, 50 and 75 mg/L) treatment caused statistically important (p < 0.05) increases of TAC levels in human lymphocytes without changing TOS levels. In conclusion, TPO can be a new resource of therapeutics as recognized in this study with its non-genotoxic and antioxidant features.

Potential anticancer activity of carvone in N2a neuroblastoma cell line.

Carvone (CVN) is a monocyclic monoterpene found in the essential oils of Mentha spicata var. crispa (Lamiaceae) and Carum carvi L. (Apiaceae) plants and has been reported to have antioxidant, antimicrobial, anticonvulsant, and antitumor activities. The beneficial health properties of CVN have encouraged us to look into its anticancer activity. To the best of our knowledge, reports are not available on the anticancer activity of CVN in cultured primary rat neuron and N2a neuroblastoma (NB) cells. Therefore, the present study is an attempt toward exploring the potential anticancer activity of CVN, if any, in cultured primary rat neuron and N2a NB cells. Our results indicated that CVN (only at 25 mg/L) treatment led to an increase in the total antioxidant capacity levels in cultured primary rat neuron cells compared with control cells. Also, CVN (at concentrations higher than 100 mg/L) treatment led to an increase in the total oxidative stress levels in both cell types. The mean values of the total scores of cells showing DNA damage (for comet assay) were not found to be significantly different from the control values in both cells (p > 0.05). On the other hand, after 24 h treatment with CVN, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay showed that CVN application significantly reduced the cell viability rates in both cell types at concentrations higher than 100 mg/L. Summarizing, our data suggest that CVN represents little potential for promising anticancer agent to improve brain tumors therapy.

The effect of carvacrol on healthy neurons and N2a cancer cells: some biochemical, anticancerogenicity and genotoxicity studies.

Carvacrol (CVC) is a phenolic monoterpene present in many essential oils of medicinal and aromatic plants and has attracted attention because of its beneficial biological activities. To date, although various biological activities of CVC have been demonstrated, its neurotoxicity on cultured primary rat neurons and N2a neuroblastoma cells has never been explored. Therefore, in this present study, we aimed to describe in vitro antiproliferative and/or cytotoxic properties (by 3-(4,5 dimetylthiazol -2-yl)-2,5 diphenlytetrazolium bromide (MTT) test), genotoxic damage potentials (by single cell gel electrophoresis (SCGE) or Comet assay) and antioxidant activities (by total antioxidant capacity (TAC) and total oxidative stress (TOS) analysis) of CVC in vitro. Dose (0-400 mg/L) dependent effects of CVC were tested on both cultured primary rat neurons and N2a neuroblastoma cells. Statistical analysis of MTT assay results indicated significant (p < 0.05) decreases of cell proliferation rates in both cell types treated with CVC at 200 and 400 mg/L. On the other hand, the mean values of the total scores of cells showing DNA damage (for comet assay) was not found significantly different from the control values for both cells (p > 0.05). In addition, our results indicated that 10, 25 and 50 mg/L of CVC treatment caused increases of TAC levels in cultured primary rat neurons but not in the N2a cell line. However, CVC treatments led to increases of TOS levels in cultured primary rat neurons at only 400 mg/L while they led to increases of TOS levels in N2a neuroblastoma cells at 200 and 400 mg/L. The present findings demonstrated that CVC could be a source of antioxidant and chemopreventive activities to be studied on cancer diseases.

The cytogenetic effects of the aqueous extracts of migratory locust (Locusta migratoria L.) in vitro.

One of the useful and most commonly cultivated commercially species, migratory locust (Locusta migratoria; Orthoptera), was investigated in light of genotoxic damage potentials. For this aim, we evaluated the genotoxic potentials of water soluble extracts of L. migratoria on cultured human blood cells. The micronucleus, sister chromatid exchange and structural chromosome aberration assays were applied to assess DNA and chromosomal damage produced by aqueous extracts in vitro. The extracts were added to the cultures at different concentrations ranging from 0 to 1000 mg/L. Our results indicated that these extracts did not exhibit genotoxicity at tested concentrations. We conclude that this in vitro approach for biomonitoring genotoxicity assessment is useful for comparing the potential health risks of edible insects.

Role of aqueous Bryoria capillaris (Ach.) extract as a genoprotective agent on imazalil-induced genotoxicity in vitro.

In recent years, a number of studies have suggested that lichens might be the easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being carried out to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. On the other hand, imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce very serious toxic effects in vertebrates. In this context, the antigenotoxic effect of aqueous Bryoria capillaris (Ach.) extract (BCE) was studied against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberrations (CA) and micronucleus (MN) as cytogenetic parameters. Human peripheral lymphocytes were treated in vitro with varying concentrations of BCE (5, 10, 25, 50 and 100 µg/mL), tested in combination with IMA (336 µg/mL). BCE alone was not genotoxic, and when combined with IMA treatment, it reduced the frequency of CAs and the rates of MN. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of BCE. The results of the present study suggest that this plant extract per se do not have genotoxic potential, but can modulate the genotoxicity of IMA on peripheral human lymphocytes in vitro. In conclusion, our findings may have an important application in the protection of cultured human lymphocyte from the genetic damage and side effects induced by agricultural and medical chemicals that are hazardous to people.

Anticancer and antioxidant properties of terpinolene in rat brain cells.

Terpinolene (TPO) is a natural monoterpene present in essential oils of many aromatic plant species. Although various biological activities of TPO have been demonstrated, its neurotoxicity has never been explored. In this in vitro study we investigated TPO's antiproliferative and/or cytotoxic properties using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) test, genotoxic damage potential using the single-cell gel electrophoresis (SCGE), and oxidative effects through total antioxidant capacity (TAC) and total oxidative stress (TOS) in cultured primary rat neurons and N2a neuroblastoma cells. Dose-dependent effects of TPO (at 10 mg L(-1), 25 mg L(-1), 50 mg L(-1), 100 mg L(-1), 200 mg L(-1), and 400 mg L(-1)) were tested in both cell types. Significant (P<0.05) decrease in cell proliferation were observed in cultured primary rat neurons starting with the dose of 100 mg L(-1) and in N2a neuroblastoma cells starting with 50 mg L(-1). TPO was not genotoxic in either cell type. In addition, TPO treatment at 10 mg L(-1), 25 mg L(-1), and 50 mg L(-1) increased TAC in primary rat neurons, but not in N2a cells. However, at concentrations above 50 mg L(-1) it increased TOS in both cell types. Our findings clearly demonstrate that TPO is a potent antiproliferative agent for brain tumour cells and may have potential as an anticancer agent, which needs to be further studied.

The genoprotective activity of resveratrol on permethrin-induced genotoxic damage in cultured human lymphocytes

The aim of this work was to investigate the genetic effects of resveratrol (RSV) at concentrations of 10, 15, 25, 40, 75 and 100 µM and its activities on the genotoxicity induced by the permethrin (PM) (200 µM). After the application of PM and RSV, separately and together, cultured human lymphocytes were assessed by chromosome aberrations (CA) and sister chromatid exchange (SCE) tests. According to results, the frequencies of CA and SCE rates in the peripheral lymphocytes were significantly increased by PM compared with the controls. However, RSV had no genotoxic effect. Furthermore, the findings revealed that PM-induced increases in the mean frequencies of both genotoxic indices were diminished by RSV in a clear dose dependent manner, indicating its protective role towards the cells from PM exerted injury. In conclusion, these effects of RSV should be considered while evaluating the possible use of RSV as a therapeutic agent.

Anti-genotoxic role of eicosapentaenoic acid against imazalil-induced DNA damage in vitro.

Eicosapentaenoic acid (EPA) is a polyunsaturated n-3 fatty acid and is essential to the health of mammals. Recent data show that EPA can act as anti-mutagenic agent. On the other hand, pesticides comprise a new and important class of environmental pollutants nowadays. Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains is suspected to produce very serious toxic effects in vertebrates. The present study investigated the anti-genotoxic effect of EPA against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberration (CA) and micronucleus (MN) tests as cytogenetic endpoints. Peripheral blood cells were treated in vitro with varying concentrations of EPA (2.5, 5, 10, 20 and 40 µg/ml), tested in combination with IMA (336 µg/ml). Our results revealed that the rates of CAs and MNs in lymphocytes were significantly (p < 0.05) increased by IMA as compared to the controls. The results also showed that EPA alone was not genotoxic. Moreover, when combined with IMA treatment, EPA reduced the frequencies of CAs and MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of EPA. In conclusion, our data may have an important application for the protection of cultured human lymphocyte from the genetic damage and repercussions induced by agricultural and industrial chemicals hazardous in people.

An antidote for imazalil-induced genotoxicity in vitro: the lichen, Dermatocarpon intestiniforme (Körber) Hasse.

Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce serious toxic effects in vertebrates. In recent years, a number of studies have suggested that lichens might be easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being performed to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. In this context, the antigenotoxic effect of aqueous Dermatocarpon intestiniforme (Körber) Hasse. extract (DIE) was studied against the genotoxic damage induced by IMA on cultured human lymphocytes (n = 6) using chromosomal aberration (CA) and micronucleus (MN) as cytogenetic endpoints. Human peripheral lymphocytes were treated in vitro with varying concentrations of DIE (0, 25, 50 and 100 µg/ml), tested in combination with IMA (336 µg/ml). DIE alone were not genotoxic and when combined with IMA treatment, it reduced the frequency of CAs and the rate of MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of DIE. The results of the present study suggest that this plant extract per se does not have a genotoxic potential, but can alleviate the genotoxicity of IMA on cultured human lymphocytes. In conclusion our findings may have an important application for the protection of cultured human lymphocyte from the genetic damage and side effects induced by medical and agricultural chemicals hazardous for people.

Xanthoria elegans (Link) (lichen) extract counteracts DNA damage and oxidative stress of mitomycin C in human lymphocytes.

Several lichen species have been used for medicinal purposes throughout the ages, and they are reported to be effective in the treatment of different disorders including ulcer and cancer. It is revealed that lichens may be easily accessible sources of natural drugs and possible food supplements after their safety evaluations. The main objective in this study was to evaluate the roles of aqueous extracts of Xanthoria elegans (at 25, 50 and 100 µg/ml) upon mitomycin C (MMC; at 10(-7) M) induced genotoxic and oxidative damages in cultured human lymphocytes. X. elegans were collected from the Erzurum and Artvin provinces (in Turkey) during August 2010. After the application of MMC and X. elegans extract (XEE), separate and together, human whole blood cultures were assessed by four genotoxicity end-points including chromosomal aberration, micronucleus, sister chromatid exchange (SCE) and 8-oxo-2-deoxyguanosine (8-OH-dG) assays. In addition, biochemical parameters [total antioxidant capacity (TAC) and total oxidative stress (TOS)] were examined to determine oxidative effects. According to our results, the frequencies of cytogenetic endpoints and 8-OH-dG levels were significantly increased by MMC compared with controls in human peripheral lymphocytes. MMC caused oxidative stress by altering TAC and TOS levels. On the contrary, XEE led to increases of TAC level without changing TOS level. XEE had no genotoxic effect. Furthermore, our findings revealed that MMC induced increases in the mean frequencies of four genotoxic indices were diminished by XEE in dose dependent manner, indicating its protective role towards cells from MMC exerted injury. In conclusion, the results obtained in the present study indicate for the first time that XEE is a potential source of natural antigenotoxicants.

The effects of taurine on permethrininduced cytogenetic and oxidative damage in cultured human lymphocytes.

Permethrin (PM) is a common pyrethroid pesticide used to control pests in agriculture, forestry, horticulture, health care, homes, and textile industry. It is confirmed as a strong mutagen in animals and humans. Taurine (TA) is an amino acid found in mammalian tissues that protects the cell against DNA damage. In this study, we investigated whether supplementation of human lymphocyte cultures with TA (in the concentrations of 25 µg mL-1, 50 µg mL-1 and 100 µg mL-1) provided any protection against PM toxicity applied in the concentration of 200 µg mL-1. Genotoxicity was assessed using the micronucleus (MN) and sister chromatid exchanges (SCE) tests. In addition, we measured the total antioxidant capacity (TAC) and total oxidative stress (TOS) levels in the plasma to determine oxidative effects. PM increased SCE and MN levels and altered TAC and TOS levels. TA alone did not affect SCE and MN levels compared to controls, regardless of the concentration applied. In addition, it increased TAC levels without changing TOS levels. Moreover, it significantly buffered the negative cytogenetic and oxidative effects induced by PM in a clear dose-dependent manner. In conclusion, this study is the first to evidence the beneficial effects of TA against PM-induced DNA and oxidative damages in vitro.

Role of Peltigera rufescens (Weis) Humb. (a lichen) on imazalil-induced genotoxicity: analysis of micronucleus and chromosome aberrations in vitro.

Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce very serious toxic effects on vertebrates. On the other hand, in recent years, a number of studies have suggested that lichens might be easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being carried out to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. In this context, the anti-genotoxic effects of aqueous Peltigera rufescens (Weis) Humb. extracts (PREs) were studied against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberrations (CAs) and micronucleus (MN) as cytogenetic parameters. Human peripheral lymphocytes were treated in vitro with varying concentrations of PREs (0, 5, 10, 25, 50 and 100 mg/L), tested in combination with IMA (336 mg/L). PREs alone were not genotoxic and when combined with IMA treatment, reduced the frequency of CAs and the rates of MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of P. rufescens extract. The results of the present study indicate that this plant extract per se do not have genotoxic potential but can minimize the genotoxicity of IMA on human lymphocytes in vitro. In conclusion our findings may have an important application for the protection of human lymphocyte from the genetic damage and side effects induced by agricultural and medical chemicals hazardous in people.

The evaluation of the genotoxic and oxidative damage potentials of Ulothrix tenuissima (Kutz.) in vitro.

Several alga species are known to produce a variety of toxic metabolites that pose a threat to aquatic organisms, animals and humans. Moreover, these metabolites have been thought to cause serious diseases including certain cancers and neurodegenerative disorders. On the other hand, Ulothrix is a genus of filamentous green algae, generally found in fresh water and marine and abundantly available in some lakes and rivers of Turkey. To our best knowledge, no study has been performed to assess the genotoxic and biochemical effects of U. tenuissima on cultured human blood cells. Therefore, in order to determine clastogenic or aneugenic effects of aqueous alga extracts the micronucleus assay was carried out. Nuclear division index (NDI) in peripheral lymphocytes was also analyzed for cytotoxicity evaluations. In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative stress (TOS)) were examined to determine oxidative effects. For this aim, we obtained heparinized blood samples from three healthy persons. The alga samples were collected from Porsuk Pond in Hasankale (Erzurum, Turkey) in summer period of the year 2010. The aqueous extracts of this species were added to cultures at different concentrations (0 to 5000 ppm) for 72 h. Our results showed that this alga did not cause any statistically important changes in the rates of studied genotoxicity endpoint. But dose-dependent alterations were observed in TAC and TOS levels and NDI rates. In conclusion, U. tenuissima was found to be non-genotoxic but caused sterility at higher concentrations due to oxidative stress.

Effects of Lichenic Extracts (Hypogymnia physodes, Ramalina polymorpha and Usnea florida) on Human Blood Cells: Cytogenetic and Biochemical Study.

Several lichen species have been used for medicinal purposes throughout the ages, and they were reported to be effective in the treatment of different disorders including tuberculosis, hemorrhoids, ulcer, dysentery and cancer. It is revealed that they may be easily accessible sources of natural drugs that could be used as a possible food supplement or in pharmaceutical industry after their safety evaluations. However, so far, the nature and/or biological roles of plenty of lichenes have not been elucidated exactly. The aim of this study was to investigate the genetic and oxidative effects of water extracts of three different lichen species; Hypogymnia physodes, Ramalina polymorpha and Usnea florida in cultured human blood cells (n = 5) for the first time. All lichen species were collected from the Erzurum and Artvin provinces (in Turkey) during August 2010. The lichen extracts were added into culture tubes at various concentrations (0 to 2000 mg/L). Chromosome aberrations (CA) and micronucleus (MN) tests were used for genotoxic influences estimation. In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative stress (TOS)) were examined to determine oxidative effects. In our in-vitro test systems, it was observed that all tested lichen extracts had no mutagenic effects on human lymphocytes. Furthermore, these extracts exhibited antioxidant properties due to the type of lichen species added to the cultures. In conclusion, these lichens can be a new resource of therapeutics as recognized in this study with their non-mutagenic and antioxidant features.