Wykrywanie roślin GMO tolerujących glifosynat z wykorzystaniem biosensora DNA

Wykrywanie roślin GMO opiera się głównie na analizie materiału genetycznego i poszukiwaniu wprowadzonych w trakcie modyfikacji fragmentów kwasów nukleinowych. Najczęściej w tym celu stosuje się metody oparte na łańcuchowej reakcji polimerazy (PCR). Jednak stale poszukuje się nowych, mniej skomplikowanych i tańszych rozwiązań. Pomocne mogą być np. elektrochemiczne biosensory DNA. Celem pracy autorów było zbudowanie i przeprowadzenie badań czujnika przeznaczonego do wykrywania sekwencji kwasów nukleinowych specyficznych dla genetycznie zmodyfikowanych roślin tolerujących glifosynat. Cecha ta uzyskiwana jest poprzez wprowadzenie do roślin genu kodującego acetylotransferazę fosfinotricyny (gen bar). Sposób działania sensora opiera się na wykrywaniu hybrydyzacji kwasów nukleinowych z wykorzysta-niem Co(bpy)33+ (tris (2,2’-bipirydylu) kobaltu(III)), który zastosowano jako elektroaktywny wskaźnik reakcji zachodzących na powierzchni elektrody z pasty węglowej. Przydatność czujnika oceniono badając próby DNA wyizolowane z tytoniu GMO tolerującego glifosynat. Odpowiednio dobrane parametry pomiarowe umożliwiły wykrycie genetycznych modyfikacji w próbach DNA o stężeniu 0,6 i 0,9 μg/ml w trakcie analizy trwającej ok. 25 minut, przy względnym odchyleniu standardowym 2-3%. Prezentowany sensor może być wykorzystany w mało kosztownej metodzie przesiewowej do wykrywania roślin GMO.Detection of GMO plants is mainly based on the analysis of genetic material and the search for nucleic acid fragments introduced during modification. Currently, polymerase chain reaction (PCR) methods are most commonly used for this pur-pose. However, new, less complicated and cost effective methods are being sought. Electrochemical DNA biosensors are part of this research. The aim of the present study was to develop a sensor designed to detection of nucleic acid sequences specific for GM plants. These plant were transformed by introducing a gene encoding phosphinothricin acetyltransferase (bar gene). The way the sensor works is based on the detection of nucleic acids hybridization using Co(bpy)33+ (tris(2,2′-bipyridyl)cobalt(III)), which was used as an electroactive indicator of reactions taking place on the surface of the carbon paste electrode. The usefulness of the developed sensor was assessed by testing DNA samples isolated from GM tobacco. Properly selected measurement parameters enabled the detection of genetic modifications in DNA samples with a concen-tration of 0.6 and 0.9 μg/ml during the analysis lasting about 25 min, with a relative standard deviation of 2-3%. The presented sensor can be used as a low cost screening tool for the detection of GM plants

THE TECHNICAL INFRASTRUCTURE AS A LOCAL DEVELOPMENT FACTOR: THE EXAMPLE OF TERESPOL COUNTY

The paper characterizes the technical infrastructure and its role in local development processes. Theoretical deliberations are broadened by the case study of Terespol county. The study examines actions taken by local authorities in the area of the technical infrastructure development. The discussion includes the summary of opinions of county council members about the selected issues related to the county technical infrastructure development. The purpose of the diagnostic survey was the collection of council members opinions about the level of infrastructure development, infrastructure role in county development, and selected problems and priorities in infrastructure development in Terespol county

Application of DNA Hybridization Biosensor as a Screening Method for the Detection of Genetically Modified Food Components

An electrochemical biosensor for the detection of genetically modified food components is presented. The biosensor was based on 21-mer single-stranded oligonucleotide (ssDNA probe) specific to either 35S promoter or nos terminator, which are frequently present in transgenic DNA cassettes. ssDNA probe was covalently attached by 5’-phosphate end to amino group of cysteamine self-assembled monolayer (SAM) on gold electrode surface with the use of activating reagents – water soluble 1-ethyl-3(3’- dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxy-sulfosuccinimide (NHS). The hybridization reaction on the electrode surface was detected via methylene blue (MB) presenting higher affinity to ssDNA probe than to DNA duplex. The electrode modification procedure was optimized using 19-mer oligoG and oligoC nucleotides. The biosensor enabled distinction between DNA samples isolated from soybean RoundupReady® (RR soybean) and non-genetically modified soybean. The frequent introduction of investigated DNA sequences in other genetically modified organisms (GMOs) give a broad perspectives for analytical application of the biosensor

Temperature Optimization by Electrochemical Method for Improving Antioxidant Compound Extraction Efficiency from <i>Origanum vulgare</i> L. and Its Application in a Bread Production

This study aims to evaluate the effect of extraction temperature on the electrochemical activity of antioxidant compounds in oregano extract and its application in a bread production. Temperature optimisation was performed by determining the electrochemical index (EI), calculated on the parameters of individual peaks observed on the square wave voltammograms (SWV). The highest value of EI (2.5758 µA/V) was observed at 85 °C for the oregano extract. The composition of several types of bread with oregano extract or dried oregano leaves was then proposed. To specify bread samples, both newly prepared and during their storage, their antioxidant properties were determined using FRAP (Ferric Reducing Antioxidant Power) and CUPRAC (Cupric Reducing Antioxidant Capacity) methods. The study revealed that the addition of extract from oregano or oregano leaves increased the antioxidant compounds content in the bread from 30% to more than 138% compared to the control bread samples. The performed sensory evaluation of the bread samples revealed their high acceptability. It was found that the stored bread with oregano leaves changed sensory qualities to a lesser extent compared to the bread with oregano extract

Temperature Optimization by Electrochemical Method for Improving Antioxidant Compound Extraction Efficiency from Origanum vulgare L. and Its Application in a Bread Production

This study aims to evaluate the effect of extraction temperature on the electrochemical activity of antioxidant compounds in oregano extract and its application in a bread production. Temperature optimisation was performed by determining the electrochemical index (EI), calculated on the parameters of individual peaks observed on the square wave voltammograms (SWV). The highest value of EI (2.5758 &micro;A/V) was observed at 85 &deg;C for the oregano extract. The composition of several types of bread with oregano extract or dried oregano leaves was then proposed. To specify bread samples, both newly prepared and during their storage, their antioxidant properties were determined using FRAP (Ferric Reducing Antioxidant Power) and CUPRAC (Cupric Reducing Antioxidant Capacity) methods. The study revealed that the addition of extract from oregano or oregano leaves increased the antioxidant compounds content in the bread from 30% to more than 138% compared to the control bread samples. The performed sensory evaluation of the bread samples revealed their high acceptability. It was found that the stored bread with oregano leaves changed sensory qualities to a lesser extent compared to the bread with oregano extract

Efficiency of Novel Antimicrobial Coating Based on Iron Nanoparticles for Dairy Products’ Packaging

The main function of food packaging is to maintain food&rsquo;s quality and safety. The use of active packaging, including antimicrobial materials, can significantly extend the shelf life of food. Many of these packaging solutions are based on the application of polymer films containing metal nanoparticles (e.g., Ag, Au, Cu) or metal oxides (e.g., TiO2, ZnO, MgO). However, the use of iron nanoparticles is rarely mentioned. In the study, polylactide (PLA) films containing zero-valent iron (ZVI) were made by casting method. Pure PLA films and PLA films with the addition of Fe2O3 were used as comparative materials. The composition and structure of ZVI/PLA films were evaluated with scanning electron microscopy. The XRD spectra performed on ZVI/PLA films confirmed the presence of iron in the packaging material and revealed their oxide form (Fe2O3). The addition of zero-valent iron in the concentration 1%, 3%, or 5% resulted in the formation of crystallographic planes measuring 40.8, 33.6, and 28.6 nm, respectively. The color and gloss of the films, and their antimicrobial activity against bacteria (Bacillus subtilis, Escherichia coli, Staphylococcus epidermidis) and fungi (Geotrichum candidum, Rhodotorula rubra) were also examined. The PLA films with addition of 3% of ZVI (w/w) inhibited the growth of all tested organisms in contrast to PLA and PLA/Fe2O3 films. The addition of ZVI to polymer matrix caused changes in its appearance and optical properties. The ZVI/PLA coating used on polyolefin film allowed to extend the shelf life of goat cheese packed in examined material to 6 weeks. Considering the antimicrobial properties of the ZVI/PLA films and PLA biodegradability the obtained material can be successfully applied in the food industry

The Genoprotective Role of Naringin

Since ancient times, fruits and edible plants have played a special role in the human diet for enhancing health and maintaining youthfulness. The aim of our work was to determine the interactions between naringin, a natural ingredient of grapefruits, and DNA using an electrochemical biosensor. Electrochemical methods allow analyzing the damages occurring in the structure of nucleic acids and their interactions with xenobiotics. Our study showed that the changes in the location of electrochemical signals and their intensity resulted from the structural alterations in DNA. The signal of adenine was affected at lower concentrations of naringin, but the signal of guanine was unaffected in the same condition. The dynamics of changes occurring in the peak height and surface of adenine related to naringin concentration was also significantly lower. The complete binding of all adenine bases present in the tested double-stranded DNA solution was observed at naringin concentrations ranging from 8.5 to 10.0 &micro;M. At larger concentrations, this active compound exerted an oxidizing effect on DNA. However, the critical concentrations of naringin were found to be more than twice as high as the dose absorbable in an average human (4 &micro;M). The results of our work might be helpful in the construction of electrochemical sensors for testing the content of polyphenols and would allow determining their genoprotective functionality

Polyphenol content and antioxidant activities of Prunus padus L. and Prunus serotina L. leaves: Electrochemical and spectrophotometric approach and their antimicrobial properties

The aim of the study was to compare the content of selected phytochemicals as well as the antioxidant and antimicrobial potential of the leaves of Prunus padus L. and Prunus serotina L., as there is very little research on this subject in the literature. Therefore, it is used to deepen knowledge on this subject. In addition, an electrochemical test was also carried out, which was not yet available for the above plants. Antibacterial studies have also been deepened to include the analysis of new strains of bacteria and fungi, which has not been studied earlier. The water extracts of P. padus using the utra-performance liquid chromatography (UPLC) system showed a higher content of both phenolic acids and flavonols (651.77b ± 18.12 mg/100 g dw for acids and 3.85b ± 0.08 mg/100 g dw for flavonols, respectively). Ferulic and p-coumaric acids were the dominant polyphenols in leaves. Extracts from P. padus showed higher activity against DPPH radical, which was 6.62b ± 0.06 mg TE/1 g dw, as well as higher antioxidant capacity, measured using 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cation radical (37.39b ± 3.81 mg TE/g dw). The higher antioxidant potential of P. padus was confirmed based on the oxidizing potentials of electroactive compounds present in them. Stronger inhibition against Enterococcus faecium and Klebsiella pneumoniae was found for P. padus, whereas P. serotina extract was more potent against Enterococcus faecium bacterium. It has been shown that P. padus can be an attractive raw material with antioxidant and antimicrobial properties that can be used on a much wider scale in food technology than its current application

Novel Drying Methods for Sustainable Upcycling of Brewers’ Spent Grains as a Plant Protein Source

Brewers’ spent grains (BSGs) are the most important by-product of the brewing industry and are rich in protein and fiber. However, abundant amounts of BSGs are discarded annually worldwide. This project aimed to employ and compare innovative drying techniques to introduce snacks with protein sources derived from leftover BSGs. This study explored the dehydration kinetics of BSGs and the effect of three different drying methods—oven drying (OD), freeze drying (FD), and vacuum microwave drying (VMD)—on their protein content and functionality. Then, an energy and exergy analysis for the drying methods was given. Accordingly, a snack product (baked chips) using the dehydrated BSGs was developed by a sensory panel study to assess consumer acceptability of the final products. It was found that the VMD process took less drying time (48 min) compared to OD (50 min), with higher effectiveness as a drying process. VMD-treated BSG also showed moderate protein functionality and the highest overall acceptability when used in baked chips. Thus, VMD might be used as a sustainable drying technology for thermal treatment and valorization of BSG. In addition to having implications for dietary health, findings can help improve the economy of the breweries and other industries that deal with the processing of grains by valorizing their process waste and contributing to sustainability.Land and Food Systems, Faculty ofOther UBCNon UBCReviewedFacult