- Email: [email protected]
Utilization of chloramphenicol imprinted nanoparticles in surface plasmon resonance nanosensors
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Abstracts / Journal of Biotechnology 231S (2016) S4–S109
Appropriate concentration of Hydrogen Peroxide and Sulforaphane for granulosa cells to study oxidative stress in vitro Mahmodul Hasan Sohel 1,∗ , Mehmet Ulas Cinar 2 , Mahmut Kalibar 2 , Korhan Arslan 3 , Serpil Sariozkan 4 , Bilal Akyuz 3 , Yusuf Konca 2 1
Genome and Stem Cell Centre, Erciyes University, Kayseri, Turkey 2 Department of Animal Science, Faculty of Agriculture, Erciyes University, Kayseri, Turkey 3 Department of Genetics, Faculty of Veterinary Medicine, Erciyes University, Kayseri, Turkey 4 Department of Reproduction and AI, Faculty of Veterinary Medicine, Erciyes University, Kayseri, Turkey E-mail address: [email protected] (M.H. Sohel). Hydrogen peroxide (H2 O2 ) and sulforaphane (SFN) are classical reagents to study oxidative stress and antioxidant activity in vitro, respectively. Micromole of H2 O2 is sufficient to induce oxidative stress. Similarly, micromole of SFN is able to induce protective activity of cells against oxidative stress. However, higher or lower concentrations of both H2 O2 and SFN may lead to aberrant results. Therefore, current study aimed to find out the appropriate concentrations of H2 O2 and SFN for granulosa cells in vitro. For this, granulosa cells were collected and cultured with DMEM-F12 media at 37 ◦ C in a humidified environment until 60–70% confluence was achieved. After that different concentrations of H2 O2 (0–400 M) and SFN (0–80 M) were added to the culture media and continue culture for 24 h. The results showed that cell viability decreased in a concentration dependent manner for both H2 O2 and SFN treated cells. 50–75 M H2 O2 are able to induce morphologically visible stress on cells, whereas 10–15 M SFN showed no effects on cellular morphology and viability. Analysis of candidate transcripts showed that, in H2 O2 groups, 50–75 M create significantly higher expression of NRF2 & CASP3 and lower expression of KEAP1. On the other hand, 10–15 M SFN exhibited significant higher expression of NRF2 and lower expression of KEAP1, while expression of CASP3 remains unchanged. It is clear from the morphologic and genetic observations that 50–75 M H2 O2 and 10–15 M SFN showed excellent response and are recommended concentrations for granulosa cells to study oxidative stress in vitro. http://dx.doi.org/10.1016/j.jbiotec.2016.05.104 Factors affecting rhamnolipid production Richard Jezdik 1,∗ , Jan Masak 2 1 Jezdik Richard, Department of Biotechnology, University of Chemical Technology Prague, Czech Republic 2 Masak Jan, Department of Biotechnology, University of Chemical Technology Prague, Czech Republic
E-mail address: [email protected] (R. Jezdik). Biosurfactants are surface active compounds with various structure produced by a wide range of microorganisms. The most known biosurfactants are glycolipids. The most studied glycolipids are rhamnolipids which are composed of one or two molecule(s) of rhamnose linked to one or two molecules of ˇ-hydroxy-fatty acid(s). The biosynthesis of rhamnolipids is mostly studied in the genus Pseudomonas. Rhamnolipids are produced as a mixture
of congeners and the highest rhamnolipid productivity occurs in the stationary phase of growth. The production of rhamnolipids is influenced by many factors, including type of production microorganism, medium composition (especially carbon and nitrogen sources) and culture conditions (pH, temperature, aeration). These factors can affect not only productivity but also the composition of rhamnolipid mixture which determines physicochemical properties of produced biosurfactant. Presented research is focused on rhamnolipid’s applicability in bioremediation processes which is dependent on the physicochemical properties of the rhamnolipid mixture. We compared several physicochemical properties of rhamnolipid mixtures produced by three different strains of Pseudomonas aeruginosa, and the effect of medium composition on rhamnolipid yield. http://dx.doi.org/10.1016/j.jbiotec.2016.05.105 Sugar beet carbon dots: Detection of Fe3+ ions Saliha Dinc Selcuk University Cumra School of Applied Sciences, Konya, Turkey E-mail address: [email protected]. Recently luminescent carbon dots, new class of carbonaceous nanomaterials, have received considerable attention. With their good water solubility, biocompatibility, low toxicity and easy synthesis, carbon dots have many application areas such as biosensor, bioimaging and photocatalysis. Electrochemical synthesis, combustion, thermal, hydrothermal, acidic oxidation and microwave are some methods are used synthesize carbon dots. In this study, carbon dots are directly extracted from sugar beet molasses, by product of refining of sugar beet factories, without using any chemical or thermal methods. Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), atomic force microscopy (AFM), UV–vis and fluorescence spectroscopy were used for the characterization of carbon dots. Sugar beet carbon dots were used as tool of biosensor in detection of Fe3+ ion. http://dx.doi.org/10.1016/j.jbiotec.2016.05.106 Utilization of chloramphenicol imprinted nanoparticles in surface plasmon resonance nanosensors Meryem Kara 1,∗ , Lokman Uzun 2 , Sevgi Kolayli 3 , Adil Denizli 2 1
Selcuk University, Advanced Technology Research and Application Center, Cumra Vocational High School, Konya, Turkey 2 Hacettepe University, Department of Chemistry, Ankara, Turkey 3 Karadeniz Technical University, Department of Chemistry, Trabzon, Turkey E-mail address: [email protected] (M. Kara). A surface plasmon resonance (SPR) nanosensor was developed for the determination of chloramphenicol (CAP) using the molecularly imprinted nanoparticles. Miniemulsion polymerization method was used to prepare the CAP imprinted nanoparticles. Non-imprinted nanoparticles were also prepared without chloramphenicol for control purpose. Then, CAP imprinted nanoparticles were immobilized to the surface of gold chip and dried in an oven at 37 ◦ C for 6 h under UV-radiation (365 nm, 100 W). Atomic force
Abstracts / Journal of Biotechnology 231S (2016) S4–S109
microscopy (AFM) and contact angle measurements were used for the surface characterization. Kinetic studies were performed with chloramphenicol solution of 0.155–6.192 nM range. Florphenicol (FLP) and thiamphenicol (TAP) antibiotics were chosen as competitive components to determine selectivity of nanoparticles. Selectivity constants were observed as 8.86 for CAP/TAP and 8.36 for CAP/FLP. The detection limit was calculated as 40 ng/kg for honey sample. http://dx.doi.org/10.1016/j.jbiotec.2016.05.107
the delivery of in vitro synthesized mRNA’s is safer than the viral vectors. Pancreatic beta cell production is necessary to create a new beta cell source for diabetes patients whose beta cells are lost due to an auto-immune attack. Some pancreatic transcription factors were shown to create beta-like insulin positive cells from somatic cells. However, the production and delivery of synthetic mRNAs of these factors have not been demonstrated yet. In this study we demonstrated that redesigned synthetic mRNAs of master pancreatic TFs can be in vitro synthesized, delivered and properly expressed in the host cells.
Determination of total phenol content, antibacterial activity and free radical scavenging activity of some Bupleurum species
http://dx.doi.org/10.1016/j.jbiotec.2016.05.109
Hatice Taner Saracoglu 1 , Ahmet Unver 2 , Mehtap Akin 1,∗
Physiological responses of two contrasting watermelon genotypes exposed to drought and nitric oxide
1
Department of Biology, Faculty of Science, Selcuk University, 42075 Konya, Turkey 2 Department of Food Engineering, Faculty of Engineering and Architecture, Necmettin Erbakan University, Konya, Turkey E-mail address: [email protected] (M. Akin). In this research, twelve Bupleurum L. (Apiaceae) species extracted by methanol–water mixture were tested for antiradical (IC50 value) potentials, and antimicrobial activities as well. The values of total phenolic content of the extracts of Bupleurum species were ranged from 56.64–317.54 mg gallic acid equivalent (GAE)/g dry extract. The free radical scavenging activity of the extracts were expressed as IC50 and the results for the samples were ranged between 0.39 and 3.41 mg/ml. In this research, the Bupleurum species extracts were tested for antimicrobial activities by microdilution method against total 11 bacterial strains: no inhibition was detected in the range of the studied extract concentrations. Bupleurum species contain some phenolic substances and have some free radical scavenging activity but, surprisingly have no inhibition effect on tested bacteria in the range of tested extract concentration. All determinations were conducted in triplicate. Data is expressed as mean. The means of results were compared by using the one-way and multivariate analysis of variance (ANOVA) followed by Duncan’s multiple range tests. The differences between individual means were deemed to be significant at P < 0.05. http://dx.doi.org/10.1016/j.jbiotec.2016.05.108 In vitro synthesis of redesigned pancreatic mRNAs: A current approach for gene delivery
Erdogan Esref Hakki 1,∗ , Mehmet Hamurcu 1 , Sait Gezgin 1 , Tijen Demiral 2 1
Department of Soil Science and Plant Nutrition, Selcuk University, Konya, Turkey 2 Department of Biology, Faculty of Science and Arts, Harran University, Sanliurfa, Turkey E-mail address: [email protected] (E.E. Hakki). Effects of nitric oxide (NO) treatment on drought tolerance of two watermelon genotypes were investigated by treating plants at three-leaf stage with 15% PEG 6000 in Hoagland solution for ten days. 100 M sodium nitroprusside (SNP) was used as NO donor. Drought stress decreased fresh and dry weights of shoots and roots and lengths of shoots of both watermelon genotypes and the rate of decreases were more prominent in drought-sensitive genotype KAR 147. Although NO treatment did not prevent drought-induced decreases in growth parameters, it prevented decrease in leaf RWC of both watermelon genotypes. NO did not show any effect on increased proline accumulation levels of both genotypes resulting from drought stress. NO treatment did not affect photosynthetic activity in drought-tolerant KAR 98 but decreased in KAR 147. RuBP regeneration capacity in KAR 98 and both RuBP regeneration capacity and carboxylation in KAR 147 were decreased. Drought stress caused also decreases in Ca content of shoot and roots and K contents of shoots of both genotypes. K content in roots of KAR 98 was decreased but K content in roots of KAR 147 did not change under drought stress. NO treatment inhibited decrease in Ca content in roots of KAR 98 caused by drought and caused more decreases in K contents of shoots and roots of KAR 147. Drought stress resulted in a decrease in Na content of shoots of KAR 147 and an increase in Na content of roots of KAR 98.
Ersin Akinci ∗ , Mehmet Yildiz, Pelin Unal Department of Agricultural Biotechnology, Akdeniz University, Antalya, Turkey E-mail address: [email protected] (E. Akinci). Direct cell reprogramming by addition of master transcription factors (TFs) to host cells is a new opportunity to reproduce the damaged or lost cells of patients. However, the way how TFs will be delivered into the host cells will decide if they can be used in clinic. The viral vectors are frequently used for gene delivery due to their high efficiency. However, the existence of viral DNA in host cells is a serious concern that restricts their use in clinic. On the other hand, in vitro synthesis of mRNA’s of TFs to be used and delivery of these mRNAs into cells was demonstrated to be as efficient as viral vectors for cell reprogramming. Even though it is laborious,
S25
http://dx.doi.org/10.1016/j.jbiotec.2016.05.110
Abstracts / Journal of Biotechnology 231S (2016) S4–S109
Appropriate concentration of Hydrogen Peroxide and Sulforaphane for granulosa cells to study oxidative stress in vitro Mahmodul Hasan Sohel 1,∗ , Mehmet Ulas Cinar 2 , Mahmut Kalibar 2 , Korhan Arslan 3 , Serpil Sariozkan 4 , Bilal Akyuz 3 , Yusuf Konca 2 1
Genome and Stem Cell Centre, Erciyes University, Kayseri, Turkey 2 Department of Animal Science, Faculty of Agriculture, Erciyes University, Kayseri, Turkey 3 Department of Genetics, Faculty of Veterinary Medicine, Erciyes University, Kayseri, Turkey 4 Department of Reproduction and AI, Faculty of Veterinary Medicine, Erciyes University, Kayseri, Turkey E-mail address: [email protected] (M.H. Sohel). Hydrogen peroxide (H2 O2 ) and sulforaphane (SFN) are classical reagents to study oxidative stress and antioxidant activity in vitro, respectively. Micromole of H2 O2 is sufficient to induce oxidative stress. Similarly, micromole of SFN is able to induce protective activity of cells against oxidative stress. However, higher or lower concentrations of both H2 O2 and SFN may lead to aberrant results. Therefore, current study aimed to find out the appropriate concentrations of H2 O2 and SFN for granulosa cells in vitro. For this, granulosa cells were collected and cultured with DMEM-F12 media at 37 ◦ C in a humidified environment until 60–70% confluence was achieved. After that different concentrations of H2 O2 (0–400 M) and SFN (0–80 M) were added to the culture media and continue culture for 24 h. The results showed that cell viability decreased in a concentration dependent manner for both H2 O2 and SFN treated cells. 50–75 M H2 O2 are able to induce morphologically visible stress on cells, whereas 10–15 M SFN showed no effects on cellular morphology and viability. Analysis of candidate transcripts showed that, in H2 O2 groups, 50–75 M create significantly higher expression of NRF2 & CASP3 and lower expression of KEAP1. On the other hand, 10–15 M SFN exhibited significant higher expression of NRF2 and lower expression of KEAP1, while expression of CASP3 remains unchanged. It is clear from the morphologic and genetic observations that 50–75 M H2 O2 and 10–15 M SFN showed excellent response and are recommended concentrations for granulosa cells to study oxidative stress in vitro. http://dx.doi.org/10.1016/j.jbiotec.2016.05.104 Factors affecting rhamnolipid production Richard Jezdik 1,∗ , Jan Masak 2 1 Jezdik Richard, Department of Biotechnology, University of Chemical Technology Prague, Czech Republic 2 Masak Jan, Department of Biotechnology, University of Chemical Technology Prague, Czech Republic
E-mail address: [email protected] (R. Jezdik). Biosurfactants are surface active compounds with various structure produced by a wide range of microorganisms. The most known biosurfactants are glycolipids. The most studied glycolipids are rhamnolipids which are composed of one or two molecule(s) of rhamnose linked to one or two molecules of ˇ-hydroxy-fatty acid(s). The biosynthesis of rhamnolipids is mostly studied in the genus Pseudomonas. Rhamnolipids are produced as a mixture
of congeners and the highest rhamnolipid productivity occurs in the stationary phase of growth. The production of rhamnolipids is influenced by many factors, including type of production microorganism, medium composition (especially carbon and nitrogen sources) and culture conditions (pH, temperature, aeration). These factors can affect not only productivity but also the composition of rhamnolipid mixture which determines physicochemical properties of produced biosurfactant. Presented research is focused on rhamnolipid’s applicability in bioremediation processes which is dependent on the physicochemical properties of the rhamnolipid mixture. We compared several physicochemical properties of rhamnolipid mixtures produced by three different strains of Pseudomonas aeruginosa, and the effect of medium composition on rhamnolipid yield. http://dx.doi.org/10.1016/j.jbiotec.2016.05.105 Sugar beet carbon dots: Detection of Fe3+ ions Saliha Dinc Selcuk University Cumra School of Applied Sciences, Konya, Turkey E-mail address: [email protected]. Recently luminescent carbon dots, new class of carbonaceous nanomaterials, have received considerable attention. With their good water solubility, biocompatibility, low toxicity and easy synthesis, carbon dots have many application areas such as biosensor, bioimaging and photocatalysis. Electrochemical synthesis, combustion, thermal, hydrothermal, acidic oxidation and microwave are some methods are used synthesize carbon dots. In this study, carbon dots are directly extracted from sugar beet molasses, by product of refining of sugar beet factories, without using any chemical or thermal methods. Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), atomic force microscopy (AFM), UV–vis and fluorescence spectroscopy were used for the characterization of carbon dots. Sugar beet carbon dots were used as tool of biosensor in detection of Fe3+ ion. http://dx.doi.org/10.1016/j.jbiotec.2016.05.106 Utilization of chloramphenicol imprinted nanoparticles in surface plasmon resonance nanosensors Meryem Kara 1,∗ , Lokman Uzun 2 , Sevgi Kolayli 3 , Adil Denizli 2 1
Selcuk University, Advanced Technology Research and Application Center, Cumra Vocational High School, Konya, Turkey 2 Hacettepe University, Department of Chemistry, Ankara, Turkey 3 Karadeniz Technical University, Department of Chemistry, Trabzon, Turkey E-mail address: [email protected] (M. Kara). A surface plasmon resonance (SPR) nanosensor was developed for the determination of chloramphenicol (CAP) using the molecularly imprinted nanoparticles. Miniemulsion polymerization method was used to prepare the CAP imprinted nanoparticles. Non-imprinted nanoparticles were also prepared without chloramphenicol for control purpose. Then, CAP imprinted nanoparticles were immobilized to the surface of gold chip and dried in an oven at 37 ◦ C for 6 h under UV-radiation (365 nm, 100 W). Atomic force
Abstracts / Journal of Biotechnology 231S (2016) S4–S109
microscopy (AFM) and contact angle measurements were used for the surface characterization. Kinetic studies were performed with chloramphenicol solution of 0.155–6.192 nM range. Florphenicol (FLP) and thiamphenicol (TAP) antibiotics were chosen as competitive components to determine selectivity of nanoparticles. Selectivity constants were observed as 8.86 for CAP/TAP and 8.36 for CAP/FLP. The detection limit was calculated as 40 ng/kg for honey sample. http://dx.doi.org/10.1016/j.jbiotec.2016.05.107
the delivery of in vitro synthesized mRNA’s is safer than the viral vectors. Pancreatic beta cell production is necessary to create a new beta cell source for diabetes patients whose beta cells are lost due to an auto-immune attack. Some pancreatic transcription factors were shown to create beta-like insulin positive cells from somatic cells. However, the production and delivery of synthetic mRNAs of these factors have not been demonstrated yet. In this study we demonstrated that redesigned synthetic mRNAs of master pancreatic TFs can be in vitro synthesized, delivered and properly expressed in the host cells.
Determination of total phenol content, antibacterial activity and free radical scavenging activity of some Bupleurum species
http://dx.doi.org/10.1016/j.jbiotec.2016.05.109
Hatice Taner Saracoglu 1 , Ahmet Unver 2 , Mehtap Akin 1,∗
Physiological responses of two contrasting watermelon genotypes exposed to drought and nitric oxide
1
Department of Biology, Faculty of Science, Selcuk University, 42075 Konya, Turkey 2 Department of Food Engineering, Faculty of Engineering and Architecture, Necmettin Erbakan University, Konya, Turkey E-mail address: [email protected] (M. Akin). In this research, twelve Bupleurum L. (Apiaceae) species extracted by methanol–water mixture were tested for antiradical (IC50 value) potentials, and antimicrobial activities as well. The values of total phenolic content of the extracts of Bupleurum species were ranged from 56.64–317.54 mg gallic acid equivalent (GAE)/g dry extract. The free radical scavenging activity of the extracts were expressed as IC50 and the results for the samples were ranged between 0.39 and 3.41 mg/ml. In this research, the Bupleurum species extracts were tested for antimicrobial activities by microdilution method against total 11 bacterial strains: no inhibition was detected in the range of the studied extract concentrations. Bupleurum species contain some phenolic substances and have some free radical scavenging activity but, surprisingly have no inhibition effect on tested bacteria in the range of tested extract concentration. All determinations were conducted in triplicate. Data is expressed as mean. The means of results were compared by using the one-way and multivariate analysis of variance (ANOVA) followed by Duncan’s multiple range tests. The differences between individual means were deemed to be significant at P < 0.05. http://dx.doi.org/10.1016/j.jbiotec.2016.05.108 In vitro synthesis of redesigned pancreatic mRNAs: A current approach for gene delivery
Erdogan Esref Hakki 1,∗ , Mehmet Hamurcu 1 , Sait Gezgin 1 , Tijen Demiral 2 1
Department of Soil Science and Plant Nutrition, Selcuk University, Konya, Turkey 2 Department of Biology, Faculty of Science and Arts, Harran University, Sanliurfa, Turkey E-mail address: [email protected] (E.E. Hakki). Effects of nitric oxide (NO) treatment on drought tolerance of two watermelon genotypes were investigated by treating plants at three-leaf stage with 15% PEG 6000 in Hoagland solution for ten days. 100 M sodium nitroprusside (SNP) was used as NO donor. Drought stress decreased fresh and dry weights of shoots and roots and lengths of shoots of both watermelon genotypes and the rate of decreases were more prominent in drought-sensitive genotype KAR 147. Although NO treatment did not prevent drought-induced decreases in growth parameters, it prevented decrease in leaf RWC of both watermelon genotypes. NO did not show any effect on increased proline accumulation levels of both genotypes resulting from drought stress. NO treatment did not affect photosynthetic activity in drought-tolerant KAR 98 but decreased in KAR 147. RuBP regeneration capacity in KAR 98 and both RuBP regeneration capacity and carboxylation in KAR 147 were decreased. Drought stress caused also decreases in Ca content of shoot and roots and K contents of shoots of both genotypes. K content in roots of KAR 98 was decreased but K content in roots of KAR 147 did not change under drought stress. NO treatment inhibited decrease in Ca content in roots of KAR 98 caused by drought and caused more decreases in K contents of shoots and roots of KAR 147. Drought stress resulted in a decrease in Na content of shoots of KAR 147 and an increase in Na content of roots of KAR 98.
Ersin Akinci ∗ , Mehmet Yildiz, Pelin Unal Department of Agricultural Biotechnology, Akdeniz University, Antalya, Turkey E-mail address: [email protected] (E. Akinci). Direct cell reprogramming by addition of master transcription factors (TFs) to host cells is a new opportunity to reproduce the damaged or lost cells of patients. However, the way how TFs will be delivered into the host cells will decide if they can be used in clinic. The viral vectors are frequently used for gene delivery due to their high efficiency. However, the existence of viral DNA in host cells is a serious concern that restricts their use in clinic. On the other hand, in vitro synthesis of mRNA’s of TFs to be used and delivery of these mRNAs into cells was demonstrated to be as efficient as viral vectors for cell reprogramming. Even though it is laborious,
S25
http://dx.doi.org/10.1016/j.jbiotec.2016.05.110