Conference MendelNet 2015      11 and 12 November, 2015
Contributions

List of Contributions Conference MendelNet 2015

Applied Chemistry and Biochemistry

  • ANTIVIRAL ACTIVITY OF FULLERENES MODIFIED WITH MAXIMIN H5 DERIVATIVES
    DOSTALOVA SIMONA, MILOSAVLJEVIC VEDRAN, GURAN ROMAN, KOMINKOVA MARKETA, CIHALOVA KRISTYNA, KOPEL PAVEL, VACULOVICOVA MARKETA, ADAM VOJTECH, KIZEK RENE
    view abstract fulltext simona1dostalova@gmail.com

    Abstract: The properties of various peptides, including their antiviral or antibacterial activity, are highly dependent on their sequence. Maximin and maximin H peptides can be abundantly found in the skin and brain of Asian frog Bombina maxima, enabling it to fight with the frequent exposure to various microbes. Derivatives of 20 amino acid long maximin H5 also show antiviral activity. In this work, 6 derivatives of maximin H5 were prepared, with aspartic acid at position 11 exchanged for alanine, asparagine, glycine, histidine, valine or tyrosine. Their antiviral and antimicrobial activity was measured using plaque assay or growth curves method, respectively. To increase these properties, the peptides were bound to C60 fullerenes, whose surface was activated using either nitric or trimesic acid. As model organisms, bacteriophage λ and its host bacteria Escherichia coli were used. The mutation of maximin H5 sequence significantly increased its antiviral activity. Maximin H5 derivatives with aspartic acid exchanged for asparagine, valine or tyrosine had the highest antiviral activity, further increased when bound on the surface of fullerenes activated with trimesic acid.  


     
  • BIOSORPTION EFFICIENCY OF CADMIUM IONS BY GREEN ALGAE (CHLOROPHYTA) IN AQUEOUS SOLUTIONS
    HYNSTOVA VERONIKA, KLEJDUS BORIVOJ, HEDBAVNY JOSEF
    view abstract fulltext veronika.hynstova@mendelu.cz

    Abstract: The aim of this study is to explore the biosorption process of Cd2+ ions by the dry algal biomass (Coccomyxa subellipsoidea, Chlorella vulgaris, Parachlorella kessleri, Scenedesmus quadricauda, Trebouxia erici)and to investigate the biosorption process by the plant biosorbent Tillandsia usneoides from aqueous solutions. In this study, we tested the effect of biosorbent dosage and contact time of Cd2+ ions with different dried algal biomass or plant on biosorption efficiency (%). The initial concentration of Cd2+ ions was 10 mg/L. The solutions were filtered in four time intervals. Final concentration of Cd2+ ions in the filtrates was determined by the atomic absorption spectrometry on the CONTRAA 700 (Analytik Jena) at the wavelength 228.8 nm. The biosorption efficiency was found to be biomass dosage dependent. When the biomass dose was increased ten times, from 0.2 g/L on 2 g/L, the maximum biosorption efficiency of Cd2+ increased 75.05% from 39.01. The alga Parachlorella was demonstrated as the most effective biosorbent of ions Cd2+ with maximum percentage biosorption 73.14% (corresponds to 3.657 mg Cd2+/g DW), in comparison to Tillandsia usneoides which only reached 37.66% (corresponds to 1.916 mg Cd2+/g DW).


     
  • AUTOMATION AND MINIATURIZATION OF BETA-GLUCOSIDASE KINETICS MEASUREMENT
    KLIMES PAVEL
    view abstract Paja.klimes@gmail.com

    Maize beta-glucosidase Zm-p60.1 is enzyme (EC 3.2.1.21) that hydrolyses various substrates to form glucose and aglycon. Plant hormones, cytokinins are released from natural conjugates and these compounds are important for the regulation of cell division and plant development. Aglycons of artificial substrates have often suitable optical properties to measure their concentrations like para-nitrophenole that can be easily measured at high pH by spectrophotometry at 405 nm. We automated and miniaturized enzyme kinetic measurements using artificial substrate para-nitrophenyl-O-beta-D-glucoside.

    All steps of enzyme kinetics were automated and scaled down to fit 384-well microtitration plate format (prefilling plate with 0.2 M Na2CO3 solution, enzyme dispensing, substrate prefilling, enzymatic reaction performing). All steps were performed by BioNex Nanodrop II liquid handling robotics placed in the incubator to secure 30 °C during enzymatic reaction. There was also a cooling position that helped to stop enzymatic reaction in the Na2CO3 solution. The procedure is highly customable (changing volumes of enzyme and substrate, reaction time etc.). Advantages of this automation setup are rapid reduction of chemical consumption and saving time due to using 8 needles that dispensed reaction mixture in parallel. Next advantage is dispensing of reaction mixture aliquots in 3 sec intervals that is not possible by hand pipetting. Results obtained from automated method were compared to hand pipetting with no significant difference in accuracy was observed.
    Keywords: enzyme kinetics, automation, pNPG, beta-glucosidase


     
  • CONSTRUCTION OF REMOTE-SENSING PLATFORM FOR STRATOSPHERIC EXPERIMENTS
    KUDR JIRI, ZITKA JAN, MILOSAVLJEVIC VEDRAN, NEJDL LUKAS, ADAM VOJTECH, KIZEK RENE
    view abstract fulltext george.kudr@centrum.cz

    Abstract: Nowadays remote sensing represents alternative to standard laboratory analysis. It also remains the only possibility to perform analysis in dangerous or for other reasons inaccessible conditions like volcanoes, atmosphere or highly contaminated areas. These devices perform in situ analysis in order to minimize processes, which can negatively influence results. Manual sample handling, sample contamination or changes during transportation are some of them. The research of atmosphere is appealing field of science. The effects of UV radiation on living organisms which is presented above the ozone layer is known for decades, nevertheless laboratory researches are not able to evaluate the additive effect of several others negative effects like low pressure, temperature or presence of highly energetic particles. The aim of our research was to create remote sensing platform, which will be able to perform several electrochemistry-based analysis in stratosphere and enable us to quantify the effect of above-mentioned conditions on DNA.


     
  • PREPARATION AND CHARACTERIZATION OF ZINC COMPLEXES AND EVALUATION OF THEIR ANTIMICROBIAL ACTIVITY
    SKALICKOVA SYLVIE, KOPEL PAVEL, CIHALOVA KRISTYNA, NEJDL LUKAS, MELROS RODRIGO MIGUEL ANGEL, SLADEK ZBYSEK, KIZEK RENE
    view abstract fulltext sylvie.skalickova@gmail.com

    Abstract: Zinc chelates with diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA) have been prepared and conditions for Zn2+ release have been studied. The prepared Zn2+ chelate complexes are of following compositions: ZnCl2(EDTA), Zn(ClO4)2·6H2O (EDTA), ZnCl2(NTA), ZnCl2(NTA)3(btc), Zn(ClO4)2·6H2O (NTA), ZnCl2 (NDA), Zn(ClO4)2·6H2O (NDA),ZnCl2(NDA)3(btc). (H3btc = 1,3,5-benzenetricarboxylic acid). All variants of Zn2+ complexes were diluted to the concentration range 0 – 1420 µM and the absorbance spectra in the range of 230 – 330 nm were measured. The antimicrobial properties of Zn2+ complexes were studied by the method of the growth curves of the bacteria cultures Staphyloccocus aureus. The 50% inhibitory concentration was determined to 500 µM of each Zn2+ complex. It has been found that the Zn2+ complexes showed increased antimicrobial effect on Staphyloccocus aureus.


     
  • TESTING SENSITIVITY OF ARABIDOPSIS CYTOKININ RECEPTOR CRE1/AHK4 EXPRESSED IN E. COLI
    TUREK DUSAN, KLIMES PAVEL, MAZURA PAVEL, SPICHAL LUKAS, BRZOBOHATY BRETISLAV
    view abstract dusanturek@seznam.cz

    Testing of biologically active compounds is always challenging task. Here we want
    to demonstrate the power of liquid handling system Nanodrop II during the testing of citokinin-like compounds. The detecting system consists of the CRE1/AHK4 receptor expressed in E. coli strain KMI001 and several tested chemicals citokinin-like compounds. E. coli and tested compounds are cultivated together for 5h in microtiter plates. When the compound is recognized
    by the CRE1/AHK4, receptors trigger the expression of a reporter gene for the enzyme beta-galactosidase. After 5h of cultivation 4-methylumbelliferyl beta-D-galactopyranoside (artificial substrate for beta-galactosidase) is added. As final step the fluorescence intensity
    of 4-methylumbelliferone (EX 365 nm, EM 448 nm) is measured using microtiter plate reader.
    The fluorescence intensity corresponds to the amount of cytokinin-like compound used in the experiment. Using automatic liquid handling system Nanodrop II resulting in reduced time needed for dispensing ligands, E. coli suspensions and substrates compared to the hand pipetting. The accuracy of the liquid handling system allows to dispense small volumes reducing
    the consumption of chemicals. The number of tested compounds in one experimental run can be increased which speeds up testing the library of compounds and decrease the experimental time.


     
  • EVALUATION OF APOPTOSIS AND NECROSIS OF PERITONEAL MACROPHAGES IN RATS AFTER INJECTION OF ZINC CHELATES INTO ABDOMINAL CAVITY
    VAVROVA EVA, SLADEK ZBYSEK, KOPEL PAVEL, KOMINKOVA MARKETA, ADAM VOJTECH
    view abstract fulltext eva.vavrova.umfgz@mendelu.cz

    Abstract: Aim of this study was to evaluate the influence of zinc chelates injected into abdominal cavity on viability of peritoneal macrophages. Three organic acids (EDTA, DTPA, NTA) were used as zinc carriers. 24 female rats were employed in this study. The rats were divided into 5 groups: 6 received intraperitoneal injection of 2 mL 40 mM Zn-EDTA (group E), 6 received intraperitoneal injection of 2 mL 40 mM Zn-DTPA (group D), 6 rats received intraperitoneal injection of 40 mM 2 mL Zn-NTA (group N), 3 rats received 2 mL of normal saline (group K4-6) and 3 rats (group K1-3 for control) were intact. On the day after injection all rats were sacrificed and peritoneal lavages were performed and cell viability analysis was done. The macrophages were divided in two morphologically different groups – group of smaller monocytes-like macrophages (ML) with kidney-shaped nuclei and pseudopodia on their surface, and a group of macrophages with spherical nuclei and many vacuoles in the cytoplasm (vacuolized macrophages, VM). Apoptosis of ML macrophages of the peritoneum in rats administrated with Zn-DTPA was almost similar to apoptosis of cells in intact animals. This means that properties of this chelate are very close to homeostasis of rats´ abdominal cavity. The apoptosis significantly increased in group E compared to K1-3. There was a significant difference between groups K1-3 and K4-6. As for necrosis the values for K1-3 and Zn-DTPA are again very close. The most damage of cells was caused by Zn-EDTA chelate. Apoptosis of vacuolized macrophages was significantly higher in groups K4-6, E and N. Necrosis of vacuolized macrophages was significantly higher in groups K4-6 and N. The Zn-DTPA chelate looks to be the mildest carrier for Zn into the organism. The present study showed that the zinc-organics acid chelates are not toxic or irritating tissues after being injected into rat´s abdomen. The Zn-DTPA had the smallest influence to the peritoneal macrophages.


     
  • THE DISTRIBUTION AND MOBILITY OF HEAVY METALS IN THE SOILS FROM DRAHANY UPLAND
    VOROS DOMINIK, GERSLOVA EVA, GERSL MILAN, ZEMAN JOSEF
    view abstract fulltext vorosdominik@gmail.com

    Abstract: The main aim of the study was to evaluate a level of contamination by selected elements (zinc, lead, copper and arsenic) in soils from the southeastern part of Drahany Upland. In total, it has been collected forty-eight topsoil samples (DV01–DV48) and sixteen surface water samples (W1–W16). The heavy metal concentration has been detected by X-ray fluorescence spectroscopy (XRF) in the topsoil samples and Atomic Absorption Spectroscopy (AAS) in the surface water samples. The heavy metal mobility has been evaluated by the BCR Sequential Extraction Procedure. The Index of Geoaccumulation (Igeo) reported that the study area rang among uncontaminated or moderately contaminated by heavy metals. According to the Coefficient of Industrial Pollution (CIP) the soils are medium contaminated. The BCR method proved that lead and zinc are the most mobile elements under reducible conditions in the natural background soils, otherwise in the soils of excepted contaminated site, heavy metals have been mostly held in residual fraction. The low mobility of cuprum and arsenic has been reported in the soils. In fact, the copper and arsenic are not supposed to being come into the soil environment and into the plants, either.