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Majer-Baranyi K.,Food Science Research Institute | Adanyi N.,Food Science Research Institute | Nagy A.,Food Science Research Institute | Bukovskaya O.,DoubleDelta Kft. | And 2 more authors.
International Journal of Environmental Analytical Chemistry | Year: 2015

A label-free, optical waveguide lightmode spectroscopy based immunosensor was developed for frog (Bombina orientalis) vitellogenin (Vtg) determination in biological samples as a biomarker for exogenous oestrogen compounds. Antibody against Vtg was produced in rabbits immunised with purified lipovitellin (Lpv), a precursor of Vtg, from the homogenised ovary of oriental fire-bellied toads (B. orientalis). The purified protein and Lpv/Vtg-specific serum were applied in both competitive and direct immunoassay formats using optical waveguide lightmode spectroscopy immunosensor. When measuring Vtg in direct manner, the Lpv antibody (1.76 µg mL−1) was immobilised on the sensor surface, and the linear measuring range for Vtg was 0.1–10 µg mL−1. During the competitive measurement, 100 ng mL−1 Lpv was applied for the immobilisation. The linear measuring range for Vtg was 0.5–50 ng mL−1. We studied the relative substrate specificity of the antibody, and it was concluded that the method is suitable for the sensitive and selective determination of Vtg levels in toads. Heart, liver and gonad samples from male animals were spiked with Vtg and were analysed using the newly developed method, and female toads and spawn samples were tested and compared to the calibration curve obtained by the spiked samples. © 2015 Taylor & Francis. Source


Seres A.,Szent Istvan University | Kiss I.,Szent Istvan University | Nagy P.,Szent Istvan University | Saly P.,Hungarian Academy of Sciences | And 2 more authors.
Plant, Soil and Environment | Year: 2014

Despite the fact that, on average, approximately 5–6 metric tons/ha of Bt maize stubble enter the soil on more than 170 million of hectares worldwide, the environmental impact of genetically modified maize plants on the arbuscular mycorrhizal fungi (AMF) is poorly known. In this study, the mycorrhizal colonisation on the roots of Bt maize (DAS-59122-7) and its near isogenic line was examined during the whole vegetation period. Cry3 toxin-producing Bt maize and its near isogenic line were grown in an experimental field in Julianna-major, Nagykovácsi, Hungary. DAS-59122-7 maize produces Cry34Ab1, Cry35Ab1 toxins and pat proteins for herbicide tolerance. The study assessed whether similar arbuscular mycorrhizal colonisation can be observed on the root of the Bt and near isogenic maize line and whether there are any differences in the temporal dynamics of AMF development. The arbuscular, hyphal and the arbuscular mycorrhizal fungi colonisation were higher in the near isogenic line as compared to its Bt counterpart, but no significant effect of the maize line was found as regards vesicle colonisation. The intensity of the arbuscular infection increased over time during plant maturation. DAS-59122-7 Bt maize had a negative effect on the initial development of AMF under field conditions, but no difference was seen in the case of the last two sampling dates (day 82 and 135). The reason of the latter is still not known. © 2014, Institute of Agricultural and Food Information. All rights reserved. Source


Varga S.,Agro Environmental Research Institute | Pathare G.R.,Max Planck Institute of Biochemistry | Pathare G.R.,Friedrich Miescher Institute for Biomedical Research | Baka E.,Agro Environmental Research Institute | And 6 more authors.
Journal of Microbiological Methods | Year: 2015

A variety of protein expression tags with different biochemical properties has been used to enhance the yield and solubility of recombinant proteins. Ubiquitin, SUMO (small ubiquitin-like modifier) and prokaryotic ubiquitin like MoaD (molybdopterin synthase, small subunit) fusion tags are getting more popular because of their small size. In this paper we report on the use of ubiquitin-like small archaeal modifier proteins (SAMPs) as fusion tags since they proved to increase expression yield, stability and solubility in our experiments. Equally important, they did not co-purify with proteins of the expression host and there was information that their specific JAB1/MPN/Mov34 metalloenzyme (JAMM) protease can recognize the C-terminal VSGG sequence when SAMPs fused, either branched or linearly to target proteins, and cleave it specifically. SAMPs and JAMM proteases from Haloferax volcanii, Thermoplasma acidophilum, Methanococcoides burtonii and Nitrosopumilus maritimus were selected, cloned, expressed heterologously in Escherichia coli and tested as fusion tags and cleaving proteases, respectively. Investigated SAMPs enhanced protein expression and solubility on a wide scale. T. acidophilum SAMPs Ta0895 and Ta01019 were the best performing tags and their effect was comparable to the widely used maltose binding protein (MBP) and N utilization substance protein A (NusA) tags. Moreover, H. volcanii SAMP Hvo_2619 contribution was mediocre, whereas M. burtonii Mbur_1415 could not be expressed. Out of four investigated JAMM proteases, only Hvo_2505 could cleave fusion tags. Interestingly, it was found active not only on its own partner substrate Hvo_2619, but it also cleaved off Ta0895. © 2015 Elsevier B.V. Source


Mortl M.,Agro Environmental Research Institute | Kereki O.,Budapest University of Technology and Economics | Darvas B.,Agro Environmental Research Institute | Klatyik S.,Agro Environmental Research Institute | And 3 more authors.
Journal of Chemistry | Year: 2016

Movement of two neonicotinoid insecticide active ingredients, clothianidin (CLO) and thiamethoxam (TMX), was investigated in different soil types (sand, clay, or loam) and in pumice. Elution profiles were determined to explore differences in binding capacity. Soil characterized by high organic matter content retained the ingredients, whereas high clay content resulted in long release of compounds. Decrease in concentration was strongly influenced by soil types: both CLO and TMX were retained in loam and clay soils and showed ready elution through sandy soil and pumice. Elution capability of the active ingredients in sandy soil correlated with their water solubility, indicating approximately 30% higher rapidity for TMX than for CLO. Soil organic carbon-water partitioning coefficients (Koc) determined were in good agreement with literature values with somewhat lower value for CLO in sandy soil and substantially higher values for TMX in clay soil. High mobility of these neonicotinoid active ingredients in given soil types urges stronger precautionary approach taken during their application. © 2016 Mária Mörtl et al. Source


Defarge N.,University of Caen Lower Normandy | Takacs E.,Agro Environmental Research Institute | Lozano V.L.,University of Caen Lower Normandy | Mesnage R.,University of Caen Lower Normandy | And 3 more authors.
International Journal of Environmental Research and Public Health | Year: 2016

Pesticide formulations contain declared active ingredients and co-formulants presented as inert and confidential compounds. We tested the endocrine disruption of co-formulants in six glyphosate-based herbicides (GBH), the most used pesticides worldwide. All co-formulants and formulations were comparably cytotoxic well below the agricultural dilution of 1% (18-2000 times for co-formulants, 8-141 times for formulations), and not the declared active ingredient glyphosate (G) alone. The endocrine-disrupting effects of all these compounds were measured on aromatase activity, a key enzyme in the balance of sex hormones, below the toxicity threshold. Aromatase activity was decreased both by the co-formulants alone (polyethoxylated tallow amine—POEA and alkyl polyglucoside—APG) and by the formulations, from concentrations 800 times lower than the agricultural dilutions; while G exerted an effect only at 1/3 of the agricultural dilution. It was demonstrated for the first time that endocrine disruption by GBH could not only be due to the declared active ingredient but also to co-formulants. These results could explain numerous in vivo results with GBHs not seen with G alone; moreover, they challenge the relevance of the acceptable daily intake (ADI) value for GBHs exposures, currently calculated from toxicity tests of the declared active ingredient alone. © 2016 by the authors; licensee MDPI, Basel, Switzerland. Source

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