Particle ID: A Multiplexed Hydrogel Bead Platform for Biomedical Applications.

We present a new platform based on hydrogel beads for multiplex analysis that can be fabricated, barcoded, and functionalized in a single step using a simple microfluidic assembly and a photo-cross-linking process. The beads are generated in a two-phase flow fluidic system and photo-cross-linking of the polymer in the aqueous phase by C,H insertion cross-linking (CHic). The size and shape of the hydrogel particles can be controlled over a wide range by fluidic parameters. During the fabrication of the beads, they are barcoded by using physical and optical barcoding strategies. Magnetic beads and fluorescent particles, which allow identification of the production batch number, are added simultaneously as desired, resulting in complex, multifunctional beads in a one-step reaction. As an example of biofunctionalization, Borrelia antigens were immobilized on the beads. Serum samples that originated from infected and non-infected patients could be clearly distinguished, and the sensitivity was as good as or even better than ELISA, the state of the art in clinical diagnostics. The ease of the one-step production process and the wide range of barcoding parameters offer strong advantages for multiplexed analytics in the life sciences and medical diagnostics.

An electrochemical immunosensor for sensitive detection of Escherichia coli O157:H7 by using chitosan, MWCNT, polypyrrole with gold nanoparticles hybrid sensing platform.

An electrochemical immunosensor for the common food pathogen Escherichia coli O157:H7 was developed. This novel immunosensor based on the PPy/AuNP/MWCNT/Chi hybrid bionanocomposite modified pencil graphite electrode (PGE). This hybrid bionanocomposite platform was modified with anti-E. coli O157:H7 monoclonal antibody. The prepared bionanocomposite platform and immunosensor was characterized by using cyclic voltammetry (CV). Under the optimum conditions, the results have shown the order of the preferential selectivity of the method is gram negative pathogenic species E. coli O157:H7. Concentrations of E. coli O157:H7 from 3×101 to 3×107cfu/mL could be detected. The detection limit was ∼30cfu/mL in PBS buffer. Briefly, we developed a high sensitive electrochemical immunosensor for specific detection of E. coli O157:H7 contamination with the use of sandwich assay evaluated in this study offered a reliable means of quantification of the bacteria. For the applications in food quality and safety control, our immunosensor showed reproducibility and stability.

The antioxidant and antigenotoxic potential of methanol extract of Cladonia foliacea (Huds.) Willd.

In this article, the genotoxic and antigenotoxic effects of methanol extract of of Cladonia foliacea (Huds.) Willd. (CME) were studied using WP2, Ames (TA1535 and TA1537), and sister chromatid exchange (SCE) test systems. The results of our studies showed that 5 µM concentration of aflatoxin B1(AFB1) changed the frequencies of SCE and malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GPx) activities. When 5 and 10 µg/mL concentrations of CME was added to AFB1, the frequencies of SCE and MDA level were decreased and SOD, GSH, and GPx levels were increased. The extract CME did not show any mutagenicity on Ames (Salmonella typhimurium TA1535, TA1537) and WP2 (Escherichia coli) test systems. On the other hand, CME has antimutagenicity on the mentioned test systems. The results of this experiment have clearly shown that CME has a significant antioxidative and antigenotoxic effect, which is thought to be due to the antigenotoxic activities of antioxidant enzymes.

Antigenotoxic potencies of a lichen species, Evernia prunastri.

In this article, the genotoxic and antigenotoxic effects of methanol extract of Evernia prunastri (Huds.) Willd. (MEP) were studied using WP2, Ames (TA1535 and TA1537) and sister chromatid exchange (SCE) test systems. The results obtained from bacterial test systems demonstrated that MEP has strong antimutagenic potencies on TA1537 and WP2 strains. The highest inhibition rates for MEP on TA1537 and WP2 strains were 37.70% and 69.70%, respectively. According to the SCE test system, MEP reduced the genotoxic effects of aflatoxin. In order to clarify the mechanism underlying the antigenotoxic effects of MEP, the antioxidants were determined. Cotreatments of 5, 10 and 20 µg/mL concentrations of MEP with aflatoxin B1 decreased the frequencies of SCE and the malondialdehyde level and increased amount of superoxide dismutase, glutathione and glutathione peroxidase which were decreased by aflatoxin. The data obtained from this work have clearly shown that MEP has significant antigenotoxic effects which are thought to be partly due to the antioxidant activities and antioxidant inducing capability of MEP. This is the first report indicating the antigenotoxic activities of MEP against several mutagen agents such as N-methyl-N'-nitro-N-nitrosoguanidine, acridin and aflatoxin.

Genotoxic, antigenotoxic and antioxidant properties of methanol extracts obtained from Peltigera horizontalis and Peltigera praetextata.

Now-a-days, there is a big need to reduce genotoxic effects of mutagenic and carcinogenic agents in environment, which are increased by the technological development. Lichens produce a wide variety of unique metabolites due to being in various extreme areas and being symbiotic organisms of fungi and algae. Therefore, this study was planned to search new sources having antimutagenic activity by researching two different lichen species and to determine whether their usage is safe. With this respect, the mutagenic and antimutagenic properties of methanol extracts of the lichens were determined by the bacterial reverse mutation and sister chromatid exchange assays. Furthermore, the malondialdehyde level, superoxide dismutase, glutathione and glutathione peroxidase activities against aflatoxin B1 were determined for understanding the ways in which the lichens showed their genotoxic properties.

Randomly amplified polymorphic-DNA analysis for detecting genotoxic effects of Boron on maize (Zea mays L.).

This study was carried out to investigate the genotoxic effect of boron (B) on maize using randomly amplified polymorphic DNA (RAPD) method. Experimental design was conducted under 0, 5, 10, 25, 50, 100, 125, and 150 ppm B exposures, and physiological changes have revealed a sharp decrease in root growth rates from 28% to 85%, starting from 25 ppm to 150 ppm, respectively. RAPD-polymerase chain reaction (PCR) analysis shows that DNA alterations are clearly observed from beginning to 100 ppm. B-induced inhibition in root growth had a positive correlation with DNA alterations. Total soluble protein, root and stem lengths, and B content analysis in root and leaves encourage these results as a consequence. These preliminary findings reveal that B causes chromosomal aberration and genotoxic effects on maize. Meanwhile, usage of RAPD-PCR technique is a suitable biomarker to detect genotoxic effect of B on maize and other crops for the future.

Genotoxic and antigenotoxic potentials of two Usnea species.

For ages, lichens have long been investigated popularly for biological roles, mainly antitumor, antimicrobial and antioxidant activities. Many positive results were obtained in these previous research. Thus, in this study, we aimed to determine whether extracts of Usnea articulata (UAE) and Usnea filipendula (UFE) possessing a protection against aflatoxin B1 (AFB1)-induced genotoxic and oxidative damage. The results of our studies showed that 5 µM concentrations of AFB1 increased the frequencies of sister chromatid exchange (SCE) and the level of malondialdehyde (MDA) and decreased the activities of superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GPx). However, when 5, 10 and 20 µg/mL concentrations of UAE and UFE was added to AFB1, the frequencies of SCE and MDA level were decreased and SOD, GSH and GPx level were increased. The Ames (Salmonella typhimurium TA1535, TA1537) and WP2 (Escherichia coli) test systems carried out evinced that UAE and UFE possess any mutagenicity, but have antimutagenic effects. Consequently, the results of this experiment have clearly shown that UAE and UFE have strong antioxidative and antigenotoxic effects that are associated with its antioxidant nature. A detailed study can be performed to determine the antioxidant properties of each compound that will extend the use of lichen extracts in food and pharmacy industries.

Determination of cytotoxic and genotoxic effects of naphthalene, 1-naphthol and 2-naphthol on human lymphocyte culture.

Naphthalene, a bicyclic aromatic hydrocarbon, has toxic effects on animals and humans. Although recent studies stressed on the genotoxic and cytotoxic effects of naphthalene and its metabolites on eukaryotic cells, there is a big controversy among the results of these studies. The aim of this study is to investigate the effects of naphthalene and its metabolites on the cytotoxicity and genotoxicity in the human lymphocytes in the culture. The genotoxic and cytotoxic effects of naphthalene and its metabolites, 1-naphthol and 2-naphthol, were studied using cytotoxicity test (lactate dehydrogenase and cell proliferation (WST-1) assays) and DNA fragmentation assay (terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay). Naphthalene and its metabolites had no significant cytotoxic effect on treated samples when compared with untreated ones. This result was also confirmed by WST-1 assay. In the TUNEL assay, DNA fragmentation was induced significantly by all concentrations of naphthalene and 2-naphthol and 50 and 100 µM concentrations of 1-naphthol (p < 0.05 or 0.001). In the DNA fragmentation, the most effective dose of 2-naphthol (63%) was 100 µM, when compared with negative control group (13%). These results suggest that naphthalene and its metabolites, 1-naphthol and 2-naphthol, may cause DNA damage on human lymphocytes.

Determination of the antigenotoxic potencies of some luteolin derivatives by using a eukaryotic cell system, Saccharomyces cerevisiae.

In this study, we aimed to examine the mutagenic and antimutagenic potencies of three luteolin derivatives (luteolin-7-O-glucoside, luteolin-7-O-rutinoside and luteolin-7-O-glucuronide) by using a eukaryotic cell system, Saccharomyces cerevisiae (RS112). In the antimutagenicity assays, these luteolin derivatives showed antimutagenic effects in deletion and intrachromosomal recombination events against ethyl methanesulfonate and acridine mutagen agents. In deletion events, the highest inhibition rates for luteolin-7-O-glucoside, luteolin-7-O-rutinoside and luteolin-7-O-glucuronide against ethyl methanesulfonate were 57.6%, 58.3% and 62.5%, respectively. Likewise, the highest inhibition rates for luteolin-7-O-glucoside, luteolin-7-O-rutinoside and luteolin-7-O-glucuronide against acridin were 21.8%, 22.4% and 23.6%, respectively. Our findings showed that these luteolin derivatives have stronger antimutagenic properties against ethyl methanesulfonate compared to the acridine mutagen agent.

Genotoxic effects of catmint (Nepeta meyeri Benth.) essential oils on some weed and crop plants.

This study investigates the genotoxicity of the essential oils extracted from the aerial parts of catmint (Nepeta meyeri Benth.) against two weeds (Bromus danthoniae and Lactuca serriola) and two crop plants (Brassica napus and Zea mays). The essential oils of N. meyeri analyzed by gas chromatography-mass spectrometry contained 14 compounds, with 4aα, 7α, 7aß-nepetalactone (83.4%), 4aα, 7α, and 7aα-nepetalactone (8.83%) as the major components. The oils were diluted (25, 50, 100, and 150 ppm) and the solutions were applied to seeds or leaves of these plants. The study compared the germination percentage and random amplified polymorphic DNA (RAPD) results with the control group. The results showed that the oils had a strong inhibitory activity and caused a change in RAPD profiles in terms of variation in band intensity, loss of bands, and appearance of new bands compared with the control group. The results suggested that RAPD analysis could be applied as a suitable biomarker assay for the detection of genotoxic effects of plant allelochemicals. This study indicates the genotoxical potential of N. meyeri essential oils on weed and crop plants.

Genotoxic and cytotoxic effects of storax in vitro.

The aim of this study is to investigate the effects of the storax balsam, which is a kind of sweet gum obtained from the Liquidambar orientalis Mill trees, on cell viability, cytotoxicity and genotoxicity in human lymphocyte in vitro. We studied the genotoxic effects of the extract of storax balsam (SE) using sister chromatid exchange (SCE) test system. Also the cytotoxic and inhibitory effects on cell proliferation of SE were evaluated using lactate dehydrogenase (LDH) assay and cell proliferation (WST-1) assay. The SCE frequency was increased when the cells were treated with 1.6 and 4.0 µg/mL SE concentrations (p < 0.05). Moreover, treatment of the cells with the same concentrations significantly depleted the cell number at 24th and 48th hours and elevated the LDH levels (p < 0.05) at 48th hour. These results suggest that SE can be used as an alternative antibacterial and antipathogenic agent due to its cytotoxic and genotoxic effects.

Assessment of total aflatoxin level in red pepper obtained from Istanbul.

Aflatoxins (Aspergillus flavus toxins (AFT)) are biologically active secondary metabolites mostly produced by some Aspergillus species that causes hepatotoxicity, teratogenicity, immunotoxicity, and cancers in human. The aim of this study is to determine the level of total AFT in powdered red pepper in the retail markets in 40 district of Istanbul using enzyme-linked immunosorbent assay. Of the 36 unpacked powdered red pepper samples, 32 samples (88%) contained AFT in the range of 0.2-106.4 µg/kg; 16 samples (44.4%) were above the regulatory limit which is at 10 µg/kg for total AFT in Turkey. More precautions on the production, transport, harvest, and storage of red pepper should be taken on hygiene to prevent toxic and carcinogenic effects of AFT.

Determination of protective role of selenium against aflatoxin B1-induced DNA damage.

Selenium is an essential mineral for a healthy life. Appropriate doses of it may undertake a protective role in the organism. In this study, the protective role of selenium (Se(4+)) against aflatoxin B1 (AFB1)-induced DNA damage was determined using random amplified polymorphic DNA on two plants including Vicia faba and Zea mays. It was observed that the concentrations of 0.1, 0.2 and 0.4 ppm of AFB1 have increased polymorphism value, total chlorophyll inhibition rate (IRc, %) and total protein IR (IRp, %). Unlike protein, chlorophyll contents and genomic template stability were decreased. With the addition of different concentrations (0.8 and 80 ppm) of Se(4+) to the treated samples with AFB1, the values return to normal. An 800-ppm concentration of Se(4+), on the contrary, could not inhibit the toxicity of AFB1 but caused an increase in toxicity level of AFB1/enhanced the toxicity level of AFB1. Results suggested that Se(4+) has an antagonistic effect against AFB1 toxicity and that the degree of antagonistic effect of Se(4+) against AFB1 was related to its concentration.

Protective role of methanol extracts of two lichens on oxidative and genotoxic damage caused by AFB1 in human lymphocytes in vitro.

In this study, the antigenotoxic and antioxidant effects of Umbilicaria vellea (UME) and Xantho somloensis (XME) extracts were determined using sister chromatid exchange (SCE), micronuclei (MN) assays, and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and malondialdehyde (MDA) levels against the effects of aflatoxin B(1) (AFB(1))-induced oxidative stress and genotoxicity in human lymphocytes in vitro. The results showed that the frequencies of SCE, MN, and MDA level decreased, but the activities of SOD and GPx increased when 5 µg/mL and 10 µg/mL doses of UME and XME were added to AFB(1)-treated cultures. Also the present results indicate that strong antioxidative and the antigenotoxicity mechanisms of UME and XME are associated with its antioxidant nature.

Anti-oxidative and anti-genotoxic effects of methanolic extract of Mentha pulegium on human lymphocyte culture.

In the present work, methanolic extract of Mentha pulegium from Erzurum, Turkey, was used in order to report the results of anti-oxidant capacity, anti-oxidant activity and anti-genotoxic effects. The total antioxidant capacity and total phenolic content were measured by using CUPRAC, ABTS and Folin-Ciocalteu colorimetric methods. The total phenolic content was higher than the total antioxidant capacity (for the results of both the CUPRAC and ABTS methods) of methanolic extract of M pulegium (ME). Also, we evaluated the anti-oxidant enzyme activity such as superoxide dismutase (SOD) and glutation peroxidase, total glutation (GSH) and malondialdehyde (MDA) in human lymphocyte culture. In CCl4-treated group, the activity of SOD, glutathione peroxidase (GPx) and GSH decreased significantly and the level of MDA increased significantly. A significant increase in the activity of SOD, GPx and the level of GSH were seen when supplemented with ME to CCl4-treated group. Furthermore, a significant decrease in the level of MDA was observed when compared with CCl4 alone treated group. In addition, anti-genotoxic effect of ME was studied by using sister chromatid exchange (SCE) method. As a result, ME has shown anti-genotoxic effect depend on anti-oxidative effect on human lymphocyte culture.

Anti-oxidative and anti-genotoxic effects of carnosine on human lymphocyte culture.

The aim of this study was to investigate the effects of carnosine, a biological antioxidant, on the oxidative stress and genotoxicity by a single dose of carbon tetrachloride (CCl(4); 5 mM) in the human lymphocyte culture. We studied the anti-genotoxic effects of carnosine by using sister chromatid exchange (SCE) test system. Also, the anti-oxidative effects of carnosine were evaluated by using superoxide dismutase (SOD), glutathione peroxidase (GPx), total glutathione (GSH) and malondialdehyde (MDA) assay. The SCE frequency was increased when treated with CCl(4). Carnosine at 10 and 20 mM reduced SCE frequency in the human lymphocyte (p < 0.001). In addition, CCl(4) treatment significantly depleted the level of GSH, reduced the activity of SOD and GPx and elevated the level of MDA (p < 0.001). Carnosine treatment led to significant attenuation of CCl(4)-induced oxidative stress by normalization of the activities of SOD and GPx and the level of GSH and MDA (p < 0.05 or 0.001). These results suggest that carnosine could provide anti-oxidative and anti-genotoxic protection for the oxidative and genotoxic agents that cause many diseases including cancer and neurodegenerative disease.

Protective role of methanol extract of Cetraria islandica (L.) against oxidative stress and genotoxic effects of AFB1 in human lymphocytes in vitro.

In this study, the antigenotoxic and antioxidant effects of Cetraria islandica methanol (CME) extract were determined by using sister chromatid exchange (SCE), micronuclei (MN) assays and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and malondialdehyde (MDA) levels against effects of aflatoxin B1 (AFB1) induced oxidative stress and genotoxicity in human lymphocytes in vitro. The results showed that the frequencies of SCE, MN and MDA level decreased, SOD and GPx activities increased when 5 µg/mL and 10 µg/mL doses of CME were added to AFB1-treated cultures. Also, the present results indicate that CME has strong antioxidative and the antigenotoxicity mechanisms of CME are associated with its antioxidant nature.

Key roles of vitamins A, C, and E in aflatoxin B1-induced oxidative stress.

Aflatoxins (Aspergillus flavus toxins) are one of the natural toxic molecules which are produced by a group of fungi called Aspergillus. Foods and drinks contaminated with aflatoxins cause global health and environmental problems. Today in many developing countries, these toxins are leading cause of some liver cancers and serious gastrointestinal problems. Aflatoxins, which are well known to be mutagenic, carcinogenic, hepatotoxic, and immunosuppressive, exert inhibitory effects on biological processes including DNA synthesis, DNA-dependent RNA synthesis, DNA repair, and protein synthesis. Aflatoxins B(1) (AFB(1)) is the most widespread oxidative agent of the aflatoxins. Numerous diverse compounds and extracts have been reported to reduce the aflatoxins induced oxidative stress in the body. Most of these inhibitors including phenylpropanoids, terpenoids, alkaloids, and vitamins are originally derived from plants. Among these, being essential biomolecules, vitamins are used as coenzymes in very significant biological reactions. They also function as nonenzymatic antioxidative agents protecting the cells from oxidative stress-induced toxicity and transformation. This chapter reviews the mechanism of AFB(1)-induced oxidative stress and focuses on the protective effects of vitamins A, C, and E on reducing this stress.

Protective effects of selenium against sister chromatid exchange induced by AFG 1 in human lymphocytes in vitro.

Aflatoxins have been shown to be hepatotoxic, carcinogenic, mutagenic and teratogenic to different species of animals. Besides, at low concentrations, Selenium (Se(4+)) is antimutagenic and anticarcinogenic while it is toxic, mutagenic and carcinogenic at high concentrations. In this study, we aimed to evaluate the effect of Se(4+) against aflatoxin GAFG(1) (AFG(1)) on blood cultures in relation to induction of sister chromatid exchange (SCE). The results showed that at 0.4 and 0.8 parts per million (ppm) concentration of AFG(1), the frequency of SCE increased in cultured human lymphocytes. When different concentration of Se(4+) (0.08 and 8 ppm) were added to AFG(1), the frequencies of SCE decreased. Howewer, when 800 ppm concentration of Se(4+) together with 0.08 ppm AFG(1) were added to cell division inhibited in the cultures. Results suggested that Se(4+) could effectively inhibit AFG(1)-induced SCE. Besides, the protective role of Se(4+) against AFG(1)-induced SCE is probably related to its doses.

The protective role of zinc and calcium in Vicia faba seedlings subjected to cadmium stress.

The aim of the present study was to evidence the possible antagonistic effect of Zinc (Zn(2+)) and Calcium (Ca(2+)) against cadmium (Cd(2+))-induced DNA damage by using random amplification of polymorphic DNA (RAPD) and metabolic activities in Vicia faba. The results showed that all doses of Cd(2+) (10( -3) M, 10(-5) M) caused an increase in polymorphism value and a decrease in genomic template stability (GTS %). In addition, when 10( -4)-10(-6) M Ca(2+), 10(-6) M Zn(2+) were added together with 10(-3) M, 10(-4) M, 10(-5) M of Cd(2+), polymorphism value decreased besides GTS, total protein and chlorophyll content increased. Results suggested that Zn(2+) and Ca(2+) have an antagonistic effect against Cd(2+). The order of the antagonisms of Ca(2+), Zn(2+) against Cd(2+) toxicity was Ca(2+) > Zn(2+). Especially, the degree of antagonistic effect of Zn(2+) against Cd(2+) is probably related to its concentration ratio.