7 TH INTERNATIONAL SCIENTIFIC CONFERENCE THE VITAL NATURE SIGN

ISSN 2335-8653 7TH INTERNATIONAL SCIENTIFIC CONFERENCE THE VITAL NATURE SIGN Abstract book 1 ISSN 2335-8653 Scientific Committee 1. 2. 3. 4. 5. 6...
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ISSN 2335-8653

7TH INTERNATIONAL SCIENTIFIC CONFERENCE THE VITAL NATURE SIGN Abstract book

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Scientific Committee 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18.

Prof. Habil. Dr. Audrius Maruška, Vytautas Magnus University, Lithuania Prof. Habil. Dr. Algimantas Paulauskas, Vytautas Magnus University, Lithuania Prof. Stellan Hjerten, Uppsala University, Sweden Prof. Hartmut Frank, University of Bayreuth, Germany Prof. Dr. Salvatore Fanali. Chromatography Institute of the CNR, Italy Prof. Dr. Jolanta Liesienė. Kaunas University of Technology, Lithuania Prof. Dr. Rimantas Venskutonis. Kaunas University of Technology, Lithuania Prof. Dr. Liudvikas Pranevičius, Vytautas Magnus University, Lithuania Prof. Dr. Vitalis Briedis, Lithuanian University of Health Sciences, Lithuania Prof. Dr. Douglas Westerlund, Uppsala University, Sweden Prof. Dr. Staffan Nilsson, Lund University, Sweden Prof. Dr. Isaak Rashal, University of Latvia, Latvia Prof. Olav Rosef, Telemark University College, Norway Prof. Dr. Ute Pyell, University of Marburg, Germany Prof. Dr. Pavel Jandera, University of Pardubice, Czech Republic Dr. Akos Vegvari. Lund University, Biochemical center, Sweden Dr. Susanne Wiedmer, University of Helsinki, Finland Dr. Anna Rocco, Chromatography Institute of the CNR, Italy

Organizing Committee Chairman/Editor: Prof. Habil. Dr. Audrius Maruška, Vytautas Magnus University, Lithuania 1. Dr. Ona Ragažinskienė. Vytautas Magnus university, Kaunas, Lithuania 2. Prof. Dr. Algirdas Raila. Aleksandras Stulginskis University, Kaunas Lithuania 3. Asoc. Prof. Dr. Saulius Šatkauskas, Vytautas Magnus University, Lithuania 4. Prof. Dr. Gintas Saulis, Vytautas Magnus University, Lithuania 5. Dr. Reda Kubiliūtė. Vilnius University, Lithuania 6. Dr. Vilma Ratautaitė, Vilnius University, Lithuania 7. Dr. Vykintas Baublys, Vytautas Magnus University, Lithuania 8. Dr. Saulius Mickevičius, Vytautas Magnus University, Lithuania 9. Tomas Drevinskas, Vytautas Magnus University, Lithuania 10. Kristina Bimbiraitė-Survilienė, Vytautas Magnus University, Lithuania 11. Mantas Stankevičius, Vytautas Magnus University, Lithuania 12. Ieva Akuneca, Vytautas Magnus University, Lithuania 13. Jurgita Mikašauskaitė, Vytautas Magnus University, Lithuania 14. Kęstutis Buivydas, Vytautas Magnus University, Lithuania 15. Dr. Violeta Bartkuvienė, Vytautas Magnus University, Lithuania 16. Academic youth society Modusas, Vytautas Magnus University, Lithuania

Abstract speech of authors were not edited © Vytautas Magnus University, 2013 BAR CODE 9772335865005

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L-1 ELECTROCHEMICAL PROCESSES OCCURRING DURING PULSED ELECTRIC FIELD TREATMENT AND THEIR CONSEQUENCES Raminta Rodaitė–Riševičienė, Jurgita Vismantaitė, Ramunė Jakubauskaitė, Rita Saulė, Viktorija S. Dainauskaitė, Gintautas Saulis L-2 SOLID STATE FERMENTATION A VALID ALTERNATIVE FOR ENVIRONMENTAL CONTROL Nicola Tiso3, Chiara Polcaro2, Emanuela Galli2, Ona Ragažinskienė4, Vilija. Snieškienė4, Antanina Stankevičienė4, Mantas.Stankevičius1, Audrius Maruška1, Tomas Drevinskas1, Violeta Bartkuvienė1, Olga Kornyšova1 L-3 ANALYTICAL-CHEMCAL, BIOCHEMICAL, AND ECO-TOXICOLOGICAL STUDIES ON THE POPULATION DECLINE OF FRESHWATER MOLLUSCS Hartmut Frank L-4 VARIATION OF VOLATILE COMPOUNDS OF CERCIDIPHYLLUM JAPONICUM SIEBOLD ET ZUCC., LIRIODENDRON TULIPIFERA L., GINKGO BILOBA L. LEAVES DURING DIFFERENT VEGETATION PERIODS Jurgita Mikašauskaitė1, Ona Ragažinskienė2, Audrius Maruška1 L-5 EFFECT OF PHENYLALANINE-ARGININE BETA-NAPHTHYLAMIDE ON MINIMAL INHIBITORY CONCENTRATION OF ANTIBIOTICS IN THE CASE OF SALMONELLA ENTERICA CELLS Valeryia Mikalayeva, Rimantas Daugelavičius L-6 FABRICATION OF AU NANOPARTICLES IN WATER BY FEMTOSECOND LASER ABLIATION AND THEIR CHARACTERIZATION R. Kubiliūtė, K. Maximova, A. Lajevardipour, J. Yong, J. S. Hartley, A. S. M. Mohsin, P. Blandin, J. W. M. Chon, A. H. A. Clayton, M. Sentis, P. R. Stoddart, A. Kabashin, R. Rotomskis, S. Juodkazis L-7 DEPENDENCE OF CHO AND DC-3F CELL ELECTROTRANSFECTION ON CONDUCTIVITY OF ELECTOPORATION MEDIUM Milda Jakutavičiūtė, Paulius Ruzgys, Saulius Šatkauskas L-8 MODELLING OF DOSES ACQUIRED IN BRACHITERAPY PROCEDURE USING PENELOPE CODE Andrius Tidikas, Diana Adlienė L-9 EVALUATION OF THE ENERGY CONVERSION FACTOR FOR THE TLD DOSIMETERS USED IN BRACHYTHERAPY Benas Urbonavičius1, Diana Adlienė1, Yana Nikolova Gluhcheva2 L-10 CHO VIABILITY AND METABOLIC ACTIVITY DINAMIC AFTER BLEOMYCIN ELECTROTRANSFER Baltramiejus Jakštys, Paulius Ruzgys, Saulius Šatkauskas L-11 ESTABLISHING ESSENTIAL ELEMENTS OF BIOANALYSIS – BIOBANKING, MASS SPECTROMETRIC TECHNIQUES AND MINIATURIZATION Ákos Végvári 3

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L-12 THE INFLUENCEOF THE SAMPLE PREPARATION OF CARROTS (DAUCUS CAROTA L. NEPTUN) ON THE ANTIOXIDANT ACTIVITY AND PHENOLIC COMPOUNDS Justina Zykevičiūtė-Laugks, prof. LaimaČesonienė L-13 THE YIELD AND COMPOSITION OF REED CANARY GRASS BIOMASS AS RAW MATERIAL FOR COMBUSTION L.Pocienė, L. Šarūnaitė, V.Tilvikienė, J.Šlepetys and Ž.Kadžiulienė L-14 OPEN CHIP SAW-MALDI MS SAMPLE HANDLING FROM FLYING CELLS TO ELECTROCUTED PROTEINS A. Jönsson1, L. Bllaci1, S. Kjellström2, S. Lemos3, L. Eliasson4, J. Liu, L. J. R. Friend5, L. Y. Yeo5 and S. Nilsson1 L-15 APPLICATION OF DEVELOPED ON-LINE CAPILLARY ELECTROPHORESIS – REACTION DETECTION METHOD Mantas Stankevičius, Audrius Maruška L-16 MINIATURE DETECTOR FOR CAPILLARY ELECTROPHORESIS Tomas Drevinskas, Audrius Maruška P-1 HIGH-VOLTAGE PULSES GENERATE ROS IN CELL-FREE MEDIA Viktorija–Skaidrutė Dainauskaitė, Raminta Rodaite–Riševičienė, Gintautas Saulis P-2 FASCIOLA HEPATICA PARASITISMS AMONG ISOLATED POPULATIONS OF SIKA DEER Justina Monika Gibiežaitė, Irma Pūraitė, Vytautas Mažeika, Algimantas Paulauskas P-3 CHROMIUM IONS RELEASED FROM THE STAINLESS–STEEL ANODE DURING HIGH-VOLTAGE PULSE CAN DECREASE CELL VIABILITY AND QUENCH FLUORESCENCE R. Jakubauskaite, L. Zelencova, A. Zaborskaja, R. Rodaite-Riševičiene, G. Saulis P-4 RACCOON DOG ECTOPARASITES AND THEIR TRANSMITTED PATHOGENS Indrė Jonauskaitė, Edita Juškaitė, Loreta Griciuvienė, Jana Radzijevskaja, Algimantas Paulauskas P-5 FLEAS (Siphonaptera) INFESTATION ON SMALL MAMMALS IN KURŠIŲ NERIJA Indrė Lipatova1, Auksė Jurevičienė1, Algimantas Paulauskas1, Vaclovas Gedminas2 P-6 BABESIA SPP. IN QUESTING TICKS AND TICKS PARASITIZING CERVIDS Irma Pūraitė, Jana Radzijevskaja, Algimantas Paulauskas, Aniolas Sruoga P-7 UDY OF NDRG2 AND TGFβ1 GENE PROMOTER METHYLATION AND PROTEIN EXPRESSION IN DIFFERENT GRADES OF BRAIN GLIOMAS Evaldas Simanavičius1,3, Laura Bakaitė1,3, Mykolas Mikučiūnas1,4, Paulina Vaitkienė1, Giedrius Steponaitis1,3, Daina Skiriutė1, Arūnas Kazlauskas1 , Arimantas Tamašauskas2 P-8 AREG PROMOTER METHYLATION IN DIFFERENT GRADES OF GLIOMA Rūta Sincevičiūtė 1.2, Paulina Vaitkienė 1 P-9 TES GENE METHYLATION IN DIFFERENT MALIGNANCY GLIOMAS Indrė Valiulytė1.2, Paulina Vaitkienė 1 4

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P-10 COMMON MERGANSER (Mergus merganser) POPULATION ANALYSIS USING MITOCHONDRIAL DNA MARKERS IN LITHUANIA Auksė Abromaitytė1, Rūta Godliauskaite1, Aniolas Sruoga1,2, Vaida Tubelytė1, Vykintas Baublys1 P-11 NUTRITIONAL AND NON-NUTRITIONAL ELEMENTS OF THE LEAVES OF HYMALAYAN BALSAM AND SMALL BALSAM SAMPLED FROM DIFFERENT LOCATIONS IN LITHUANIA Eglė Aleksaitė, Irma Šliumpaitė, Lina Zybartaitė, Rasa Jankauskaitė, Algimantas Paulauskas, Eugenija Kupčinskienė P-12 INFLUENCE OF CADMIUM IONS ON THE REDUCED GLUTATHIONE AND LIPID PEROXIDATION IN THE LIVER AND RED BLOOD CELLS OF MICE Kristina Čijauskaitė, Rasa Bernotienė, Leonid Ivanov P-13 MOLECULAR DETECTION AND CHARACTERIZATION OF ANAPLASMA PHAGOCYTOPHILUM STRAINS IN IXODIDAE TICKS AND RESERVOIR HOST Edita Juškaitė, Algimantas Paulauskas, Jana Radzijevskaja Irma Pūraitė P-14 DEVELOPMENT OF A MIXED-MODE SOLID PHASE EXTRACTION FOR A MULTIRESIDUE ANALYSIS OF 15 BENZODIAZEPINES IN BLOOD BY RAPID GAS CHROMATOGRAPHY WITH NEGATIVE-ION CHEMICAL IONIZATION MASS SPECTROMETRY Nerijus Karlonas a, b, Arunas Ramanavicius c, Almira Ramanaviciene a P-15 FLUORESCENCE OF ETHIDIUM BROMIDE IN SUSPENSIONS OF JURKAT AND MX1 CELLS Mantas Riauka, Rimantas Daugelavičius P-16 PHYLOGENETIC ANALYSIS OF VIBURNUM (VIBURNUM) BASED ON MICROSATELLITE MARKERS S.Bogačiovienė1, J.Žukauskienė1*, A.Paulauskas1, Eugenija Kupčinskienė1, K.Senavaitytė1, L.Česonienė2, R.Daubaras2 P-17 GENETIC DIVERSITY APODEMUS SYLVATICUS AND APODEMUS FLAVICOLLIS IN LITHUANIA Karolis Sivickis, Algimantas Paulauskas, Vaclovas Gedminas P-18 GENETIC DIVERSITY OF IMPATIENS GLANDULIFERA IN SOME NORTHERN EUROPE SITES ESTIMATED BY ISSR MARKERS Kristė Stravinskaitė, Lina Zybartaitė, Judita Žukauskienė, Algimantas Paulauskas, Eugenija Kupčinskienė* P-19 ANALYSIS OF GENETIC VARIABILITY OF BLUEBERRIES (VACCINIUM L.) CULTIVARS USING MICROSATELLITE MARKERS S.Bogačiovienė1, J.Žukauskienė1*, A.Paulauskas1, Eugenija Kupčinskienė1, I.Šarauskienė1, L.Česonienė2, R.Daubaras2 P-20 RESEARCH ON TERRESTRIAL TESTATE AMOEBA (TESTACEA) IN LITHUANIA Šatkauskienė Ingrida, Štuopytė Gertrūda, Šidlauskaitė Lina, Kantautė Vaiva,

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P-21 THE EFFECTS OF LEAD AND NICKEL IONS ON TOTAL PROTEINS AND METALLOTHIONEINS SYNTHESIS IN MICE LIVER Indrė Šveikauskaitė, Jurgita Šulinskienė, Ilona Sadauskienė, Leonid Ivanov P-22 INVESTIGATION OF NITROGEN CONCENTRATION IN HYPERICUM MACULATUM CRANTZ POPULATIONS IN SOME BALTIC SITES Indrė Užubalytė, Irma Šliumpaitė, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė P-23 GENETIC ANALYSIS OF IMPATIENS GLANDULIFERA POPULATIONS USING ISSR MARKERS Justina Vaitkevičiūtė, Lina Zybartaitė, prof. Eugenija Kupčinskienė, prof. Algimantas Paulauskas P-24 SCREENING OF EPIPHYTIC PINE BACTERIA FOR ANTAGONISTIC PROPERTIES Viktorija Vaštakaitė, Odeta Buzaitė P-25 GENETIC DIVERSITY OF LITHUANIA POPULATIONS OF COMMON JUNIPER (JUNIPERUS COMMUNIS L.) EVALUATION ACCORDING TO ISSR MARKERS Alma Vilkaitytė, Ramūnas Vilčinskas, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė* P-26 HYPOCHLOROUS ACID CREATES PORES IN THE PLASMA MEMBRANE OF HUMAN ERYTHROCYTES WITH THE SIZE CLOSE TO THE SIZE OF ELECTRICAL PULSE-GENERATED PORES J. Vismantaitė, R. Rodaitė–Riševičienė, G. Saulis P-27 PHYTOTOXICITY OF AMIDOSULFURON (SULFONYLUREAS HERBICIDE) TO AQUATIC MACROPHYTE LEMNA MINOR Jūratė Žaltauskaitė 1, Roberta Norvilaitė 2 P-28 RAPID ANALYSIS OF GENETIC DIVERSITY AMONG LITHUANIA POPULATION OF IMPATIENS NOLI TANGERE L. Julija Petkevičiūtė, Lina Zybartaite, Ramūnas Vilčinskas, Algimantas Paulauskas, Eugenija Kupcinskiene P-29 DIVERSITY AND SEASONAL VARIATIONS IN PERIPHYTON COMPOSITION IN FRESHWATER ECOSYSTEMS Rasa Glasaitė, Ingrida Šatkauskienė P-30 LOW UV-B RADIATION AND SUBSTRATE ACIDITY IMPACT ON PHOTOSYNTHEIC PARAMETERS OF BRASSICA NAPUS Irena Januškaitienė P-31 MEASUREMENT OF NITROGEN DIOXIDE CONCENTRATION IN COLD AND WARM SEASONS USING A PASSIVE SAMPLING METHOD Dovilė Laurinavičienė, Audrius Dėdelė P-32 SOIL NET CO2 EXCHANGE RATE RESPONSE TO DIFFERENT TILLAGE AND FERTILIZATION SYSTEMS Agnė Putramentaitė1, Mantas Butrimavičius2, Simona Pranaitienė3

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P-33 DISEASES OF MEDICINAL PLANTS IN KAUNAS BOTANICAL GARDEN OF VYTAUTAS MAGNUS UNIVERSITY IN SPRING SEASONS Vilija Snieškienė, Antanina Stankevičienė, Ona Ragažinskienė P-34 BIOMONITORING OF URBAN AIR QUALITY IN KAUNAS, LITHUANIA, USING TRANSPLANTED LICHENS Gintarė Sujetovienė P-35 VARIATION OF FLAVONOIDS AND PHENOLIC ACIDS IN FRUITS OF GENUS SORBUS L. PLANTS Kristina Gaivelytė, Valdas Jakštas, Valdimaras Janulis P-36 NATURAL VEGETABLE OILS POTENT ANTIOXIDANTS FOR NATURAL COSMETIC Giedrė Kasparavičienė, Dovilė Bajoriūnaitė, Saulė Velžienė, Zenona Kalvėnienė P-37 ANTIFUNGAL ACTIVITY OF PHENOLIC COMPOUNDS FROM FAGOPYRUM ESCULENTUM AGAINST FUSARIUM CULMORUM Ilona Kerienė1, Audronė Mankevičienė1, Bronislava Butkutė1, Audrius Maruška2 P-38 DETERMINATION OF PHENOLIC AND ANTIOXIDANT PROFILES OF LINDEN (TILIA CORDATA MILL.) INFLORESCENCES Raimondas Raudonis1,2*, Lina Raudonė1, Valdimaras Janulis1, Pranas Viškelis2 P-39 BOTANICAL COMPONENTS AND RAW MATERIALS OF ANIMAL ORIGIN IN THE PRESCRIPRION BOOK FROM 1830 Zenona Šimaitienė, Vilma Gudienė, Tauras Mekas P-40 SUSCEPTIBILITY OF E. COLI TOWARDS CHITOSAN MODIFIED BY SPECIFIC INORGANIC AND ORGANIC BIOCIDES Iglė Vepštaitė1,2, Juliana Lukša1, Ona Gylienė3, Rima Binkienė3, Elena Servienė1,2 P-41 BORRELIA SPP. GENOTYPING IN SMALL RODENTS AND TICKS Baliutytė Vaida, Radzijevskaja Jana P-42 INTRODUCTION AND PHYTOCHEMICAL ANALYSIS OF VIOLA TRICOLOR L. GROWN AT KAUNAS BOTANICAL GARDEN OF VYTAUTAS MAGNUS UNIVERSITY Béquet Karla 1,2,3, Audrius Maruška3, Ona Ragažinskienė2 P-43 COMPARATIVE DEVELOPMENT AND EVALUATION OF BAIKAL SCULLCAPS (SCUTELLARIA BAICALENSIS GEORGI) TOPICAL GELS, GEL – CREAMS AND CREAMS FORMULATIONS Vytis Čižinauskas, Vitalis Briedis, Kristina Ramanauskienė

P-44 DIFFERENT GROWTH RESPONSES OF PISUM SATIVUM AND MELILOTUS ALBA TO 700 AND 1500 ppm OF CARBON DIOXIDE Austra Dikšaitytė, Irena Januškaitienė P-45 BOUND STATES OF FEW NUCLEON SYSTEMS Saulius Mickevičius 7

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P-46 EFECTIVE CALCULATION OF HARMONIC OSCILLATOR TRANSFORMATION BRACKETS S. Mickevičius P-47 EVALUATION OF SOLVENT INFLUENCE OF THE QUALITY OF ETHANOLIC EXTRACTS OF SAMBUCUS NIGRA L FRUITS Saulė Velžienė, Lauryna Pudžiuvelytė, Giedrė Kasparavičienė, Zenona Kalvėnienė P-48 THE APPLICATION OF ADMS-URBAN MODEL TO ESTIMATE NITROGEN DIOXIDE CONCENTRATIONS IN COLD AND WARM SEASONS IN KAUNAS CITY Audrius Dėdelė, Auksė Miškinytė P-49 EXTRACTION PROCEDURE OPTIMIZATION OF ROSELLE (HIBISCUS SABDARIFFA L.) CALYCES Gintarė Sokaitė1, Zenona Kalvėnienė1, Giedrė Kasparavičienė1, Audrius Maruška2 P-50 ANTIOXIDANT PROPERTIES AND ELEMENT CONCENTRATIONS OF THE FLOWERS OF IMPATIENS GLANDULIFERA Eglė Kazlauskaitė, Irma Šliumpaitė, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė P-51 GLUTAMIC ACID LEVELS IN MEAT PRODUCTS K. Barčauskaitė1, G. Garmienė1, I. Miliauskienė1, I. Jasutienė1, A. Šalaševičienė1 P-52 ANISE HYSSOP INTRODUCTION AND THIS MEDICINAL PLANT RAW MATERIAL IMPORTANCE FOR HUMAN HEALTH Jovita Bagočiūtė, Mantas Stankevičius, Kristina Bimbiraitė-Survilienė, Olga Kornyšova, Vilma Kaškonienė, Ona Ragažinskienė, Audrius Maruška P-53 ANALYSIS OF HUMULUS LUPULUS L. ESSENTIAL OILS AND BITTER ACIDS Paulius Orentas1, Žydrūnas Stanius1, Paulius Kaškonas2, Tomas Drevinskas1, Mantas Stankevičius1, Kęstutis Obelevičius3, Ona Ragažinskienė3, Audrius Maruška1 P-54 VARIATION OF TOTAL AMOUNT OF PHENOLIC AND VOLATILE COMPOUNDS OF SATUREJA HORTENSIS L. AND SATUREJA MONTANA L. EXTRACTS DURING DIFFERENT VEGETATION PERIODS Andrius Rimovas1, Ieva Akuneca1, Mantas Stankevičius1, Olga Kornyšova1, Ona Ragažinskienė2, Audrius Maruška1 P-55 COMPARATIVE ANALYSIS OF RED AND WHITE WINES USING SPECTROPHOTOMETRIC AND CHROMATOGRAPHIC METHODS Raimonda Adomaitytė, Ieva Akuneca, Mantas Stankevičius, Audrius Maruška P-56 ANALYSIS OF DIFFERENT GEOGRAPHIC ORIGIN BEE-COLLECTED POLLEN USING SPECTROPHOTOMETRIC AND CHROMATOGRAPHIC TECHNIQUES Geralda Ruočkuvienė1, Vilma Kaškonienė1, Olga Kornyšova1, Audrius Maruška1 P-57 SELF-GROWTH OF GOLD-POLYPYRROLE NANOCOMPOSITES Reda Kubiliūtė1, Justas Svirelis1, Lina Mikoliūnaitė1, Jaroslav Voronovič1, Anton Popov1, Almira Ramanavičienė1,2, Arūnas Ramanavičius1,3 8

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P-58 HERBAL MEDICINES TO PREVENT NERVOUS SYSTEM DISEASES Laimutė Jonaitienė, Edita Babonienė, Ona Ragažinskienė P-59 MEDICINAL RAW MATERIAL DRYING TECHNOLOGY AND ITS BIOLOGICALLY ACTIVE SUBSTANCES EVALUATION Algirdas Raila1, Egidijus Zvicevičius1, Aurelija Kemzūraitė1, Aušra Čiplienė1, Audrius Maruška3, Ona Ragažinskienė2, Olga Kornyšova3, Vilma Kaškonienė3, Violeta Bartkuvienė3, Kristina Bimbiraitė-Survilienė3, Mantas Stankevičius3, Tomas Drevinskas3 P-60 ANALYSIS OF PHYTOCHEMICAL COMPOSITION AND ANTIBACTERIAL ACTIVITY IN MEDICINAL PLANT MYRRHIS ODORATA L. Mickienė Ruta1, Friese Anika3, Rosler Uwe3, Maruška Audrius1, Ragažinskienė Ona2 P-61 INVESTIGATION OF BIOREMEDIATION OF CREOSOTE IMPREGNATED WOODEN RAILWAY SLEEPERS J. Mikašauskaitė1, N. Tiso1, V. Snieškienė2, A. Stankevičienė2, M.Stankevičius1, A. Maruška1, T. Drevinskas1, M. Kazlauskas1, V. Bartkuvienė1, O. Kornyšova1, O. Ragažinskienė2, D. Levišauskas3, P-62 QUANTITATIVE AND QUALITATIVE ANALYSIS OF POLYPHENOLIC AND VOLATILE COMPOUNDS IN PROPOLIS USING SPECTROPHOTOMETY, LIQUID CHROMATOGRAPHY AND GAS CHROMATOGRAPHY - MASS SPECTROMETRY METHODS Giedrė Blažukienė, Vilma Kaškonienė, Loreta Kubilienė, Audrius Maruška P-63 ANALYSIS OF ESSENTIAL OILS AND PHENOLIC COMPOUNDS OF POTENTILLA FRUCTICOSA L. USING CHROMATOGRAPHIC AND SPECTROPHOTOMETRIC TECHNIQUES Šarūnas Paulaitis, Ieva Akuneca, Mantas Stankevičius, Ona Ragažinskienė, Audrius Maruška P-64 ANTIOXIDANT ACTIVITY OF DIFFERENT BOTANICAL AND GEOGRAPHICAL ORIGIN HONEY Inga Šikštėnienė, Vilma Kaškonienė, Olga Kornyšova, Audrius Maruška P-65 EVALUATION OF PHENOLIC COMPOUNDS IN APPLE (MALUS DOMESTICA BORKH.) FRUITS Mindaugas Liaudanskas1, Raimondas Raudonis1, Valdas Jakštas1, Pranas Viškelis2, Darius Kviklys2, Juozas Lanauskas2, Nomeda Kviklienė2, Valdimaras Janulis1 P-66 INVESTIGATION OF GENETIC DIVERSITY OF IXODES RICINUS TICKS BASED ON MITOCHONDRIAL DNA AND MICROSATELLITE MARKERS Banaitytė Asta, Radzijevskaja Jana, Paulauskas Algimantas, Galdikaitė Eglė

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L-1 ELECTROCHEMICAL PROCESSES OCCURRING DURING PULSED ELECTRIC FIELD TREATMENT AND THEIR CONSEQUENCES Raminta Rodaitė–Riševičienė, Jurgita Vismantaitė, Ramunė Jakubauskaitė, Rita Saulė, Viktorija S. Dainauskaitė, Gintautas Saulis Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, Kaunas, Lithuania

Electroporation is an effective method for the modification of the cell membrane permeability with an increasing number of applications in biology, oncology, genetics, immunology, and biotechnology. When using high-voltage electrical pulses for treatment of cells and tissues, scientists must keep in mind that the products generated due to electrochemical reactions can influence various processes taking part in their experimental systems. Due to this, the efficiency of electroporation technique depends not only on the parameters of electric treatment but on the electrochemical processes as well. The primary anodic and cathodic electrochemical reactions and secondary processes that can take place during the treatment by high-voltage electrical pulses used in electroporation experiments are reviewed. These include pH changes (acidification around the anode and alkalinisation at the cathode), the production of H2, O2, and Cl2 gases, and the release of the metal ions from the electrodes. Consequences of these reactions which may be important for optimization of cell electromanipulation procedures, such as arcing, the complexation of the metal ions released from the electrodes with the molecules present in the solution, quenching of fluorescence of fluorescent dyes, reduction of the cell viability, the increase of the roughness of the electrode surface due to the dissolution of the electrode material, and other ones are discussed. Keywords: cell electroporation, electrochemical reactions, hypochlorous acid, reactive oxygen species.

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L-2 SOLID STATE FERMENTATION A VALID ALTERNATIVE FOR ENVIRONMENTAL CONTROL Nicola Tiso3, Chiara Polcaro2, Emanuela Galli2, Ona Ragažinskienė4, Vilija. Snieškienė4, Antanina Stankevičienė4, Mantas.Stankevičius1, Audrius Maruška1, Tomas Drevinskas1, Violeta Bartkuvienė1, Olga Kornyšova1 1

Vytautas Magnus University (VMU), Biochemistry and Biotechnologies department, Vyleikos 8, LT-44404, Kaunas, Lithuania 2 National Research Council (CNR), Istitute of Agroenvironmental and Forestal Biology (IBAF) & Institute of Chemical Methodologies (IMC), Area dellaRicerca di Roma, Via Salaria Km 29,300, 00015, Monterotondo (Roma), Italy 3 University of Tuscia, previous Agrobiology and AgrochemistryDepartment (DABAC),Via S.C. De Lellissnc, 01100,Viterbo, Italy 4 Vytautas Magnus University Botanical garden, Ž. E. Žilibero g. 6, Kaunas, Lithuania

Abstract: Recently the use of microorganisms, such as white-rot fungi, through Solid State Fermentation (SSF) processes has been seriously investigated. SSF has shown much promise in the development of several bioprocesses and products. White rot fungi seem to be particularly interesting for environmental applications, due to their ability totolerate and/or degrade high concentrations of pollutants for their chemical transformation into less hazardous compound. The wood preservativeshave been extensively used in the last few decades to enhance the life expectancy of woody materials for external uses against natural and biological decay, but the popular waste disposal options available for these expended materials nowadays, such as burning and land filling, are not acceptable due to economical, healthand environmental concerns. The development of alternative and environmental friendly methods for disposal of these waste materials is of major importance. This presentation discusses a novel approach for bioremediation of creosote-treated railways wood sleepers by Solid State Fermentation of white-rot fungi and the possibility to combine it sequentially with phytoremediation. Keywords: Bioremediation, phytoremediation, creosote, Solid State Fermentation, white-rot fungi, wood sleepers.

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L-3 ANALYTICAL-CHEMCAL, BIOCHEMICAL, AND ECOTOXICOLOGICAL STUDIES ON THE POPULATION DECLINE OF FRESHWATER MOLLUSCS Hartmut Frank Environmental Chemistry & Ecotoxicology, University of Bayreuth, Hugo-Rüdel Str. 8, D-95445 Bayreuth, Germany [email protected]

The worldwide decline of freshwater molluscs, in particular of freshwater pearl mussels (Margaritifera margaritifera), is associated with profound alterations of the physical and chemical parameters of wetland and riverine habitats. The multitude of such impacts, like hydrological interventions, changes in land use patterns, and many secondary, biogeochemical effects, are aggravated by anthropogenic xenochemicals, especially persistent organic pollutants and certain metals. Due to industrial emissions and corrosive release associated with the use of technical products in which they are employed, the aeolian distribution of metals by particulate matter, their deposition and eventual storage in the sediments of brooks and rivers, and their transfer through the benthic food chains, certain metals are likely to be involved in the decline. Usually, toxicity tests with freshwater mussels have been carried out at fairly high concentrations and for short time periods. Through the availability of sensitive analytical methods and the use of isotopic tracers, the pathobiochemical potential of Cd and Cu could be investigated at environmentally relevant levels. Mussels incorporate them with the food and from the water. The main organs involved are the digestive gland, the gills, the mantle, and the kidney. In this process, the digestive gland acts as temporary store, while the kidney is the final sink. Here, as well as in the mantle tissue, interference with Ca2+-homeostasis is likely to be an important primary pathophysiological effect. Detailed analysis of the sequence of events of this eco-toxicological phenomenon of population decline of molluscs is needed in order to conceive protection strategies for particularly sensitive species such as freshwater pearl mussels. University Bayreuth, D-95440 Bayreuth, Germany

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L-4 VARIATION OF VOLATILE COMPOUNDS OF CERCIDIPHYLLUM JAPONICUM SIEBOLD ET ZUCC., LIRIODENDRON TULIPIFERA L., GINKGO BILOBA L. LEAVES DURING DIFFERENT VEGETATION PERIODS Jurgita Mikašauskaitė1, Ona Ragažinskienė2, Audrius Maruška1 1

Vytautas Magnus University, Faculty of Natural Sciences, Department of Biochemistry and Biotechnologies, Vileikos 8, Kaunas, LT-44404, Lithuania 2 Kaunas Botanical garden of Vytautas Magnus University, Žilibero 6, Kaunas, LT-46324, Lithuania

Cercidiphyllum japonicum Siebold et Zucc., Liriodendron tulipifera L. and Ginkgo biloba L. are woody, introduced plants in Lithuania. In this study we investigated volatile compounds present in these plants leaves during different vegetation periods. Essential oils of leaves of these plants were extracted by solid-phase microextraction (SPME), in-tube extraction (ITEX) and directly injected from headspace (HS) and analysed by gas chromatography coupled to mass spectrometry (GC–MS), to determine the chemical composition of the volatile fraction. The SPME technique was more sensitive than static headspace for analysis of plant leaves volatile compounds with GC-MS system. Comparing all three sample preparation techniques and injection techniques the SPME showed the highest concentrations, the highest number and variation of the volatile compounds, than HS or ITEX technique. Composition and amounts of volatile compounds depend on the plant, vegetation period and sample preparation for analysis. Broadest variety and concentration of volatile compounds of analysed plants, was in Cercidiphyllum japonicum leaves. Keywords: Cercidiphyllum japonicum, Liriodendron tulipifera, Ginkgo biloba, volatile compounds, GC/MS, HS, SPME, ITEX.

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L-5 EFFECT OF PHENYLALANINE-ARGININE BETA-NAPHTHYLAMIDE ON MINIMAL INHIBITORY CONCENTRATION OF ANTIBIOTICS IN THE CASE OF SALMONELLA ENTERICA CELLS Valeryia Mikalayeva, Rimantas Daugelavičius Department of Biochemistry and Biotechnologies, Vytautas Magnus University Vileikos 8, LT-44404 Kaunas, Lithuania

Resistance to antibiotics is the very common problem in medicine. It is important to understand the mechanisms of multidrug resistance in bacteria. We focused our study on efflux pump system using method of microdilutions. We used Salmonella enterica SL1344 (WT), L664 and ΔacrB strains to determine influence of phenylalanine-arginine β-naphthylamide (PAβN) on MICs of ampicillin, chloramphenicol and tetracycline. PAβN is very popular inhibitor of RND-type efflux pumps. It is toxic for bacterial cells only at high concentrations. We showed that low inactive concentrations of antibiotics can inhibit bacterial growth in combination with PAβN. Our results indicate that inhibitory activity of PAβN depends on the initial bacterial cell concentration. Such results suggest intensive binding of PAβN by the cells.

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L-6 FABRICATION OF AU NANOPARTICLES IN WATER BY FEMTOSECOND LASER ABLIATION AND THEIR CHARACTERIZATION R. Kubiliūtė, K. Maximova, A. Lajevardipour, J. Yong, J. S. Hartley, A. S. M. Mohsin, P. Blandin, J. W. M. Chon, A. H. A. Clayton, M. Sentis, P. R. Stoddart, A. Kabashin, R. Rotomskis, S. Juodkazis 1

Centre for Micro-Photonics and Industrial Research Institute Swinburne, Faculty of Engineering and Industrial Sciences Swinburne University of Technology, Hawthorn, VIC, 3122, Australia 2 Laboratory of Biomedical Physics,Vilnius University Institute of Oncology, Baublio 3A, Vilnius, LT08406, Lithuania 3 (UMR 7341 CNRS), Aix-Marseille University, Campus de Luminy-case 917, 13288, Marseille Cedex 9, France 4 The Australian National Fabrication Facility - ANFF, Victoria node, Faculty of Engineering and Industrial Sciences, Swinburne University of Technology, Hawthorn, VIC, 3122, Australia

Colloidal nanoparticles of noble metals have promising applications in sensing, nanophotonics, and optofluidics [1]. Apart of chemical methods of nanoparticle synthesis, laser ablation fabricated [2] or modified [3] nanoparticles of different metals and other materials are gaining interest due to potentially lower surface contamination, better stability against agglomeration, and simplicity of fabrication. Femtosecond laser ablation has been used to make gold nanoparticles in deionized water. Colloidal nanoparticles free of surface chemicals, usually used for stabilization, were characterized optically and structurally. Fluorescence quenching behavior and its intricacies were revealed by fuorescence lifetime imaging microscopy (FLIM) in rhodamine 6G water solution. We show that surface enhanced Raman scattering (SERS) of rhodamine 6G on gold nanoparticles can be detected with high fidelity down to micromolar concentrations using the nanoparticles. SERS applications will benefit from a wider choice of available surface modifications compatible with aqueous solutions in bio- and medical applications. Acknowledgements: RK acknowledges support via a fellowship funded by European Union Structural Funds project „Postdoc- toral Fellowship Implementation in Lithuania“ within the framework of the Measure for Enhancing Mobility of Scholars and Other Researchers and the Promotion of Student Research (VP1-3.1-ŠMM-01) of the Program of Human Resources Development Action Plan. The project was in part supported by the Australian Research Council DP120102980 project, VP1-3.1-ŠMM-01-V-02-001 funding scheme, and French National Research Agency (ANR) grants LASERNANOBIO (ANR-10-BLAN-919) and BIONANOPLASMON (ANR-09- BLAN-0177). References 1. I. M. White, S. H. Yazdi, and W. W. Yu 2012 (online published). 2. A. V. Kabashin, \Laser ablation-based methods for nanostructuring of materials," Laser Physics 19(5), pp. 1136-1141, 2009. 3. A. Pyatenko, M. Yamaguchi, and M. Suzuki, \Mechanisms of size reduction of colloidal silver and gold nanoparticles irradiated by Nd:YAG laser," J. Phys. Chem. C 113, pp. 9078-9085, 2009.

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L-7 DEPENDENCE OF CHO AND DC-3F CELL ELECTROTRANSFECTION ON CONDUCTIVITY OF ELECTOPORATION MEDIUM Milda Jakutavičiūtė, Paulius Ruzgys, Saulius Šatkauskas Vytautas Magnus University, Faculty of Natural Science, Biology Departament, Vileikos 8, Kaunas

Electroporation is a process when cell membrane permeability is increased after transmembrane voltage is lifted with external electrical fields. Thereafter exogenous molecules can penetrate through cell membrane. When plasmid DNA is electrotansfered and migrates to nucleus cell become electrotransfected. The aim of this study was to evaluate CHO and DC3F cell electrotransfection efficiency dependence on electroporation media conductivity. CHO and DC-3F cell suspension with luciferase coding plasmid (LUC) was electroporated with 1200 V/cm 100 µs various number pulses at 1 Hz in SMEM (1.3 S/m) and EP (0.1 S/m) electroporation media. Results showed better CHO electrotransfection with low conductivity electroporation medium. Opposite results were obtained with DC-3F cell line electrotransfection. Cell size and the concentration of charged carriers are proposed as an explanation to electrotransfection differences on CHO and DC3F cell lines.

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L-8 MODELLING OF DOSES ACQUIRED IN BRACHITERAPY PROCEDURE USING PENELOPE CODE Andrius Tidikas, Diana Adlienė Kaunas University of Technology, Physics Department, Studentų str. 50, LT-51368 Kaunas

Dose simulation method is usually applied in medical field in order to find out dose distributions in brachyterapy procedures. It is so, because conventional dosimetric methods not always have capacity to provide scientifically sufficient information e.g. dose measurements have limited range as well as are taking place in fixed positions and there is always possibility to have unsuccessful measurement. Because of violent nature of ionizing radiation measurements can’t be taken again. In this work, program PENELOPE-2005 is used. It is based on Monte Carlo method. Simulation geometry is made of spherical radioactive Iridium 192 source and surrounding space made of water. 370 keV monoenergetic spectres is attached to radioactive source. Dose distributions are acquired in planes near to the source. Water is selected because its properties correspond well with properties of tissue. Investigation of simulation accuracy is also being conducted. In this case we are performing theoretic calculations as well as analyzing simulation results. Both theoretical and experimental graphs are confirming same principle within the limits of errors.

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L-9 EVALUATION OF THE ENERGY CONVERSION FACTOR FOR THE TLD DOSIMETERS USED IN BRACHYTHERAPY Benas Urbonavičius1, Diana Adlienė1, Yana Nikolova Gluhcheva2 1

Kaunas University of Technology, Physics Department, Studentų str. 50, LT-51368 Kauna Plovdiv University Paisii Hilendarski, Tsar Asen St. 24, 4000 Plovdiv, Bulgaria

2

Abstract In vivo dosimetry in brachytherapy is one of the most challenging procedures in the praxis of experimental patient dosimetry. Due to the specific brachytherapy treatment procedure it is possible to insert the dosimeters via catheters directly into the cancerous tissues or into the body cavities in the nearest vicinity of the organs and tissues of interest. In this paper we provide some results on the dose evaluation in High Dose Rate (HDR) interstitial brachytherapy of Head and Neck cancer patients using TLD-100 micro dosimeters. This type of brachytherapy treatment is based on the patient‘s irradiation when the radioactive Ir192 source moves inside of the 6F catheters preimplanted in the tumor and stops at the prescribed positions to irradiate the tumor. An extra empty F6 catheter was implanted into tumor volume for dosimetric purposes. Dosimeter read outs were corrected by the assigned individual calibration factors and experimental doses were calculated. Since the calibration was performed in terms of air KERMA, obtained value was converted to absorbed dose in water. To account for difference in energy (0.37 MeV for Ir-192 and 1.25 MeV for calibration with Co-60 source) the energy conversion factor was used. Theoretical dose values corresponding to the dosimeter placements within the organ/tissue were calculated using dose planning system. Evaluated dose discrepancies are discussed on the basis of the obtained results.

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L-10 CHO VIABILITY AND METABOLIC ACTIVITY DINAMIC AFTER BLEOMYCIN ELECTROTRANSFER Baltramiejus Jakštys, Paulius Ruzgys, Saulius Šatkauskas Vytautas Magnus University, Faculty of Natural Science, Biophysical Research Group, Vileikos 8, Kaunas

Cell membrane permeability to various exogenous molecules can be enhanced when transmembrane potential is increased by exposing cells to external electric fields. This method is called electroporation. Electroporation allows intracellular delivery of various nonpermeant exogenous molecules, including anticancer drugs. Combined with anticancer drug bleomycin a new cancer treatment therapy called electrochemotherapy has been developed. To evaluate effective concentration for cell treatment with electrochemotherapy in vitro several cell viability tests are used. The aim of this study is to evaluate CHO cell death dynamics induced by cell treatment with bleomycin-based electrochemotherapy in vitro. Bleomycin (20 nm) was electrotransfered in CHO cells with one HV (1200 V/cm 100 µs) electric pulse. Metabolic activity and cell viability was measured with MTT and crystal violet tests 6, 24, 48, 72 hours after bleomycin electrotransfer. Results showed bleomycin electrotransfer significant effect on cell viability after 48 hours though significantly distinct metabolic activity was obtained after 72 hours. The results showed that cell viability after bleomycin-based electrochemotherapy can vary significantly using different test and viability evaluation time.

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L-11 ESTABLISHING ESSENTIAL ELEMENTS OF BIOANALYSIS – BIOBANKING, MASS SPECTROMETRIC TECHNIQUES AND MINIATURIZATION Ákos Végvári Lund university, Sweden

Successful analyses of biological, especially clinical, samples rely on collections in biobanks as well as on efficient analytical techniques. The quality of biobanking material has to be retained high, which requires introduction of control methods. Furthermore, the analytical power of mass spectrometric techniques provides new possibilities to screen through large sample cohorts, identifying novel biomarkers and quantifying these markers in clinical samples. One of the most important goals in the near future is to transfer these results to clinical practice, providing improved diagnostic tools for heterogeneous diseases like cancer. In this presentation, the concept and implementation of high-density biobanking, multiple reaction monitoring mass spectrometry for targeted quantitative proteomics, MALDI mass spectrometry imaging of drug localization in tissue sections and microfluidic devices designed for acustophoretic separation of cells are presented and exemplified.

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L-12 THE INFLUENCEOF THE SAMPLE PREPARATION OF CARROTS (DAUCUS CAROTA L. NEPTUN) ON THE ANTIOXIDANT ACTIVITY AND PHENOLIC COMPOUNDS Justina Zykevičiūtė-Laugks, prof. LaimaČesonienė AlexandrasStulginskisuniversity, Enviromental and ecology institute, Studentu 11, Kaunas, Lithuania

Abstract The growing demand for organic food in the world requires the assessment of the value aspects of food quality, its safety, nutritional content and biological-physiological process. When methodologically flawed studies are screened out and a complete assessment of nutritional quality is carried out, collectively, the available evidence supports the hypothesis that organically produced food is superior in terms of safety, nutritional content and nutritional value to that produced nonorganically. Carrot breeds Neptunwas chosen for the experiment from Lithuania Horticulture institute. Correlations between antioxidant activity and phenolic compounds in three drying method for sample extractions were determined. Fruits and vegetables are good sources of natural antioxidants, containing many different antioxidant components. These antioxidants include carotenoids, vitamins, phenolic compounds, flavonoids, dietary glutathione, and endogenous metabolites, having been shown to function as singlet and triplet oxygen quenchers, free radical scavengers, peroxide decomposers, enzyme inhibitors, and synergists. Phenolic compounds are found in most fruits and vegetables.Carrots containing mainly hydroxycinnamic acids and derivatives. Among them chlorogenic acid is a major hydroxycinnamic acid, representing from 42.2% to 61.8% of total phenolic compounds [1]. Aim of research was to determine antioxidant activity (DPPH), total phenolic compounds and evaluate sample preparation of carrots by using three drying methodsand different ratio (75% and 100%) of methanol for carrots samples extractions (freeze-dried under vacuum condition by -72ºC, second sliced with blender and the third was pressed carrots by the juice press, then desiccated in the fruit and vegetable desicator +40ºC). Radical scavenging activity of methanolic extracts of carrots against stable DPPH (2,2-diphenyl-2picrylhydrazyl hydrate) was determined spectrophotometricallyby slightly modified method of Brand-Williams et al.[2] and The content of phenolic compounds was determined by FolinCiocalteu colorimetric method (Folin-Ciocalteu, 1927)[3]. References 1. Donglin Zhang and YasunoriHamauzuPhenolic compounds and their antioxidant properties in different tissues of carrots (Daucus carota L.); 2. Brand-Williams, Cuvelier, and Berset (1995); 3. Wu Cu, Murthy HN, Hahn EJ, Paek KY. PMID: 17879746 [PubMed - indexed for MEDLINE]

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L-13 THE YIELD AND COMPOSITION OF REED CANARY GRASS BIOMASS AS RAW MATERIAL FOR COMBUSTION L.Pocienė, L. Šarūnaitė, V.Tilvikienė, J.Šlepetys and Ž.Kadžiulienė Institute of Agriculture, Lithuanian Research Centre for Agriculture and Forestry Instituto al. 1, LT-58344 – Lithuania Phone +37065420788 Email: [email protected]

Abstract Biomass has variable composition and properties which depends from fertilization intensity. Reed canary grass (Phalaris arundinacea L.) was investigated in small-plots experiment on light soil Arenosols. The biomass yield data in the average of two years of sward use showed that biomass yield of reed canary grass without nitrogen in this experiment was significant lower than that yield fertilized with nitrogen, and ash content ranged depends from fertilization. In comparison with grass fertilized with N120 and N0, the biomass with nitrogen fertilization N120 has shown lowest concentration of ash, which is favorable feature for combustion process. Further research on reed canary grass management is needed. Keywords: Phalaris arundinacea L., biomass yield, bioenergy potential

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L-14 OPEN CHIP SAW-MALDI MS SAMPLE HANDLING FROM FLYING CELLS TO ELECTROCUTED PROTEINS A. Jönsson1, L. Bllaci1, S. Kjellström2, S. Lemos3, L. Eliasson4, J. Liu, L. J. R. Friend5, L. Y. Yeo5 and S. Nilsson1 1

Pure & Applied Biochemistry LTH, Lund University, Sweden Department of Biochemistry and Structural Biology, Lund University, Sweden 3 Department of Biochemistry I - Ruhr University, Bochum, Germany 4 Clinical Sciences Malmö CRC, Entr 72, building 91, level 11, Malmö, Sweden 5 RMIT University, GPO Box 2476,Melbourne VIC 3001, Australia 2

Abstract SAW atomizer has been interfaced with MALDI MS for fast analysis of small volumes (< 1µL) sampled in a membrane1. SAW propagate through and underneath the membrane and atomize the liquid bound sample into ca 2 µm diameter aerosol which is subsequently deposited on a MALDI plate for further analysis2. Fast peptide profiling of living islets of Langerhans and bio fluid (saliva, tear film) has been achieved and an adaptation for the analysis of living agarose-entrapped cancer cells is currently under development. MALID MS of a single beta cells and a single islet using the acoustic levitation technique3 leads to a wall less analysis with low contamination and high sensitivity4. Electrocution of protein in conjunction with a MS interface, will be discussed as well. MALDI plate Pinhole SPUDT

SAW

Membrane SAW atomizer

Sampling volumes (< 1µL) Pre-purification Dilution/desalting Gingival fluid

Saliva

Islet of Langerhans

Tear film

Preliminary results, together with a representation of the basic SAW-MALDI sample preparation method. References 1. Bllaci, L., Kjellström, S., Eliasson, L., Friends, J.R., Yeo, L. Y., Nilsson, S., Anal. Chem. 85 (2013) 2623– 2629 2. Ho, J.T., M. K., Go, D. B., Yeo, L. Y., Friend, J. R., Chang, H.-C., Anal. Chem. 83 (2011) 260–3266. 3. Lemos, S., Gustavsson. N., Jönsson, A., Eliasson, L., Bllaci, L., Nilsson, S., 2013 (Submitted). 4. Santesson, S., Degerman, E., Rorsman, P., Johansson, T., Lemos, S., Nilsson, S., Integrative Biology 1 (2009) 595-601.

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L-15 APPLICATION OF DEVELOPED ON-LINE CAPILLARY ELECTROPHORESIS – REACTION DETECTION METHOD Mantas Stankevičius, AudriusMaruška Department of Biochemistry and Biotechnologies, Vytautas Magnus University,Vileikos 8, LT44414, Kaunas, Lithuania [email protected] Abstract There are numerous methods developed for evaluation of antioxidants in medical herbs, food products, even living cells, thus making difficult to select the most suitable for particular analysis. Determination of radical scavenging activity using synthetic radicals (2,2-diphenyl-1picrylhydrazyl, 2,2-azinobistiazolinsulfonic acid) is a relatively simple way to analyze biologically active antioxidant components in different matrices. Colored radicals with unpaired electron, are reduced while in contact with antioxidant, thus bleaching or changing the color, which can be measured with UV-Visible light detectors. Capillary electrophoresis as separation technique can be employed for separation of analytes within short time and low cost. It is a micro scale analysis, which requires minute amount of sample and omits time consuming sample preparation step, except appropriate filtration and dilution. Capillary electrophoresis coupled on-line with reaction detection, trough manually designed postcapillary reaction interface, can be applied for separation of phenolic compounds and evaluation of their radical scavenging activity in natural products and medical herbs. Capillary electrophoresis – reaction detection method was used for evaluation of antiradical properties of phenolic compounds in real samples: honey bee propolis and anise hyssop (Agastache foeniculum L.). Method showed good repeatability, low determination concentration limits, good resolution and efficiency.

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L-16 MINIATURE DETECTOR FOR CAPILLARY ELECTROPHORESIS Tomas Drevinskas, Audrius Maruška Vytautas Magnus University, Faculty of Natural Sciences, Dept. of Biochemistry and Biotechnologies, Vileikos 8, LT-44404 Kaunas

Common problem of miniaturized chemical analysis instrumentation is a bulky size of analysis handling and detection devices comparing it to the scale of microfluidic devices that usually perform the whole separation process. The aim of this work was the design of detector with only one integrated circuit. AD7745 capacitance-to-digital converter chip was used as detector in capillary electrophoresis separations. Obtained results demonstrate sensitive detector response: separated sodium ion and histidine analytes showed 5.76 µM and 8.14 µM detection limits respectively. Although, very low power requiring, miniature and novel detection technique showed peak splitting effect that can be related to inappropriate electrode geometry (conventional contactless conductivity detection electrode geometry was used). Current results and hardware issues that lead to further studies of this novel detection technique will be presented. We acknowledge the financial support of Vytautas Magnus University, project No. BF-1208.

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P-1 HIGH-VOLTAGE PULSES GENERATE ROS IN CELL-FREE MEDIA Viktorija–Skaidrutė Dainauskaitė, Raminta Rodaite–Riševičienė, Gintautas Saulis Department of Biology, Faculty of Nature Sciences, Vytautas Magnus University, Kaunas, Lithuania

Electroporation has an increasing number of applications in biology, oncology, genetics, immunology, and biotechnology. However, when cell suspension is exposed to high–voltage electric pulses, various electrochemical reactions occur at each electrode–solution interface. These may include evolution of gas, dissolution of the electrodes, pH changes, electrodissociation of molecules, and formation of reactive oxygen species (ROS). Reactive oxygen species such as superoxide radical, hydrogen peroxide, singlet oxygen and hydroxyl radical are well known to be cytotoxic and have been implicated of a wide array of human diseases, including cancer. Studying of the electrochemical processes, which occur during high–voltage pulses and their consequences, is necessary, as they are important for optimizing practical applications. In this study, it has been shown, that during high–voltage electric pulses, ROS are generated. In cell–free media, micro–millisecond pulsed electric field increased fluorescence of APF (3’-pAminophenyl fluorescein) dye emission proportionally to the pulse number and amplitude. APF fluorescence was reduced by both vitamin C and mannitol. Also, it has been shown that ROS formation was more intensive in the case of stainless–steel electrodes comparing to the aluminium ones. The results of this work can be useful for optimizing the electroporation technology used in biotechnology, medicine, and food industry. Keywords: cell electroporation, reactive oxygen species, cytotoxicity.

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P-2 FASCIOLA HEPATICA PARASITISMS AMONG ISOLATED POPULATIONS OF SIKA DEER Justina Monika Gibiežaitė, Irma Pūraitė, Vytautas Mažeika, Algimantas Paulauskas Faculty of Natural Sciences, Vytautas Magnus University, Vileikos 8, LT-44404 Kaunas, Lithuania Corresponding author. E-mail: [email protected]

Sika deer (Cervus nippon) was first brought to Lithuania in 1954 from the former Soviet Union, from the area well known as Gorno Altaisky. In Lithuania, every year more and more farmers are determined to grow cervids. Harvested deer often travel to new emerging economies, exchanged breeders. In year 2012, there were around 150 deer farms with about 2,000 sika deer held in Lithuania in them. Lithuania sika deer breeding have been started hoping to sell the horns of the calf. Now part of the farmed sika deer are beeing killed to fulfill the business needs. There was an infection registered of sika deer with plain and roundworms in reindeer famrs. Young deer thought to be very prone to dangerous Fasciola hepatica which damages the liver, bile duct, gall bladder and shows some damaging symptoms on a pancreas. In this study, 14 individuals from Kaunas region deer farms were tested for their capability to catch the infection from a lancet liver fluke. In these deer farm, there were some cases in the past when young deer were killed by Fasciola hepatica during a winter. Out of the 14 sika deer tested, all the deer were infected with Fasciola hepatica. Parasites were identified by morphology and measure. It was found that in the liver of one individual, the number of parasites ranged from 1 to 58 individuals. Prevalance of infection was 100% (80.7-100). Mean abundance ± SD = 19.0 ± 19.3. Sika deer, which were particularly strongly infected with Fasciola hepatica, were extremely gaunt and had a dilapidated coat. The deer that were the most often infected were 2-3 years of age.

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P-3 CHROMIUM IONS RELEASED FROM THE STAINLESS–STEEL ANODE DURING HIGH-VOLTAGE PULSE CAN DECREASE CELL VIABILITY AND QUENCH FLUORESCENCE R. Jakubauskaite, L. Zelencova, A. Zaborskaja, R. Rodaite-Riševičiene, G. Saulis Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, Kaunas, Lithuania

Electroporation is an effective method for the modification of the cell membrane permeability with numerous applications in biology, biotechnology, medicine, and technology. When a high–voltage pulse is applied to the electrolyte solution, besides a cell membrane permeabilization, a variety of electrolysis reactions occur at the electrode–solution interfaces. These may include evolution of gas, separation of substances, dissolution of the electrodes, and appearance of new substances. One of the most popular materials utilized for electrodes, which are used to electroporate cells, is stainless– steel. In such a case, iron, chromium, nickel, and manganese ions are released from the anode under the action of high-voltage electric pulses. These ions can affect various cell physiological processes as well as decrease cell viability. To study cell electroporation, entrance into or efflux out of the cell of fluorescent dyes is often monitoried. Metal ions can also react with fluorescent dyes and quench the intensity of their fluorescence. This may have an influence considering electroporation effectiveness results. In the release of chromoium ions from the stainless-steel anode and its consequences have been studied. It has been shown, that due to the action of a single square–wave electric pulse with the duration of 2 ms and the amplitude of 1.2 kV/cm, the concentration of chromium ions in cell electroporation media has reached 2 mg/l. The reduction of the viability of mouse hepatoma MH– 22A and rat glioma C6 cells by Cr6+ ions has also been demonstrated. These ions quench the fluorescence of calcein dye, which is used in cell electroporation studies. The results of this work can be useful for optimizing the electroporation technique used in biotechnology, medicine, and food industry. Keywords: cell electroporation, electrochemical reactions, metal ions, medium cytotoxicity.

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P-4 RACCOON DOG ECTOPARASITES AND THEIR TRANSMITTED PATHOGENS Indrė Jonauskaitė, Edita Juškaitė, Loreta Griciuvienė, Jana Radzijevskaja, Algimantas Paulauskas Faculty of Natural Sciences, Vytautas Magnus University, Vileikos 8, LT- 44404 Kaunas, Lithuania

The raccoon dog (Nyctereutes procyonoides), also known as mangut, is an invasive Canidae family mammal whose introduction from East Asia to Europe countries has been accompanied by intensive migration of these animals and spread throughout most of Europe including Lithuania. Nyctereutes procyonoides is host of various ectoparasites such as ticks, lice, and fleas, which can also carry a variety of disease-causing pathogens: viral, bacterial, and protozoan origin. Therefore it is especially important that these animals could spread pathogens that are not specific to the local fauna while migrating. In the present study ticks removed from raccoon dogs were examined for presence of different pathogens (Babesia spp., Bartonella spp., Borrelia burgdorferi sensu lato, Rickettsia spp. and Anaplasma phagocytophilyum). The possible role of raccoon dogs in spread of tick-borne transmitted diseases in Lithuania was determined. In total 44 ticks (40 Ixodes ricinus and 4 Dermacentor reticulatus) from 9 raccoon dogs have been tested by using molecular methods. We used different molecular markers, conventional and nested PCR, and sequence analysis for determination and identification of pathogen species and strains. In this study was detected Babesia microti in I. ricinus ticks from one raccoon dog. Two Rickettsia spp. species were identified: Rickettsia helvetica in I. ricinus ticks from three raccoon dogs and Rickettsia monacensis in I. ricinus tick from one raccoon dog. Bartonella spp. was found in I. ricinus from one specimen of raccoon dog. Anaplasma phagocytophilum was detected in I. ricinus harbored by one raccoon dog. Three Borrelia species were identified: one raccoon dog harbored both, B. afzelii and B. myiamotoi infected I. ricinus, and one B. valaisiana infected I. ricinus ticks. This study shows that the raccoon dogs can be implicated in transmission of a variety of pathogens that cause infections in humans and domestic animals. Financial support for this study was provided by the Research Council of Lithuania (grant.No.LEK14/2012). Keywords: raccoon dog, ticks, ectoparasites, pathogens

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P-5 FLEAS (Siphonaptera) INFESTATION ON SMALL MAMMALS IN KURŠIŲ NERIJA Indrė Lipatova1, Auksė Jurevičienė1, Algimantas Paulauskas1, Vaclovas Gedminas2 1

Department of Biology, Vytautas Magnus University, Vileikos 8, LT-44404 Kaunas, Lithuania. Kaunas Tadas Ivanauskas Zoological Museum, Laisvės 106, Kaunas, LT-44253, Lithuania E-mail: [email protected], [email protected] 2

Small mammals are vectors of ectoparasites and reservoir of various pathogens of diseases. One of rodents and insectivores ectoparasites are fleas – vectors of bacteria, viruses, cestodes, nematodes and other disease agents. In our study small mammals were captured from 2011 years to 2012 years in Kuršių Nerija. A total were captured 318 rodents (including 186 Apodemus flavicollis, 5 Apodemus sylvaticus, 22 Myodes glareolus, 5 Microtus agrestis, 14 Microtus arvalis, 7 Microtus oeconomus, and 79 Micromys 30inutes) and 48 insectivores (Sorex 30inutes). One hundred eighty four fleas (including 4 genus: Hystrichopsylla, Ctenophthalmus, Palaeopsylla, Megabothris) were collected from 128 small mammals. Small mammals’ infestations with fleas in Kuršių Nerija were 35 % (128/366). The overall prevalence of infestation with fleas was 64% for M. arvalis, 60% for A. sylvaticus, 57% for M. oeconomus, 50% for M. glareolus, 38% for M. 30inutes, 34% for A. flavicollis and 0% for M. agrestis. Fleas’ infestations on rodents were 38 % (121/318) and on insectivores were 15 % (7/48). Keywords: rodents, insectivores, infestation, fleas

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P-6 BABESIA SPP. IN QUESTING TICKS AND TICKS PARASITIZING CERVIDS Irma Pūraitė, Jana Radzijevskaja, Algimantas Paulauskas, Aniolas Sruoga Faculty of Natural Sciences, Vytautas Magnus University, Vileikos 8, LT-44404 Kaunas, Lithuania, e.mail: [email protected]

Babesiosis is an emerging, tick-transmitted, zoonotic disease caused by hematotropic parasites of the genus Babesia. Babesial parasites are some of the most ubiquitous and widespread blood parasites in the world, second only to the trypanosomes, and consequently have considerable worldwide economic, medical, and veterinary impact [1]. Human babesiosis, primarily caused by B.microti in the USA and by B.divergens in Europe was first documented in 1957 in former Yougoslavia. Since then, several hundred clinical cases in the USA and about 40 clinical cases in Europe have been recorded. However, in Europe it occurs almost exclusively in splenectomized or otherwise immunosuppressed patients [1,2]. The aim of this study was investigate the presence of Babesia parasites in questing ticks, ticks parasitizing cervids and characterize Babesia species in Lithuania. Ixodes ricinus and Dermacentor reticulatus ticks were collected from vegetation in deer farms and from cervids: red deer and roe deer in 2011-2012 years. A total 135 ticks were screened for the presence of DNA of Babesia spp. GenBank analysis of the Babesia spp. positive samples revealed presence of B.microti and B.venatorum. References 1. Homer MJ, Aguilar-Delfin I, Telford SR 3rd, Krause PJ, Persing DH. Babesiosis. Clin Microbiol Rev. 2000 Jul;13(3):451-69. 2. Gray J, Zintl A, Hildebrandt A, Hunfeld KP, Weiss L: Zoonotic babesiosis: Overview of the disease and novel aspects of pathogen identity. Ticks Tick-borne Dis 2010, 1:3-10.

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P-7 UDY OF NDRG2 AND TGFβ1 GENE PROMOTER METHYLATION AND PROTEIN EXPRESSION IN DIFFERENT GRADES OF BRAIN GLIOMAS Evaldas Simanavičius1,3, Laura Bakaitė1,3, Mykolas Mikučiūnas1,4, Paulina Vaitkienė1, Giedrius Steponaitis1,3, Daina Skiriutė1, Arūnas Kazlauskas1 , Arimantas Tamašauskas2 1Laboratory of Neurooncology and Genetics, Neuroscience Institute, Lithuanian University of Health Sciences, Eiveniu str. 4, LT 50009 Kaunas, Lithuania 2 Lithuanian University of Health Sciences, Eiveniu str. 4, LT 50009, Kaunas, Lithuania 3 Department of Biology, The Faculty of Natural Sciences, Vytautas Magnus University, Vileikos str. 8, LT 44404, Kaunas, Lithuania 4 Faculty of Medicine, Lithuanian University of Health Sciences, Mickevičiaus str. 9, LT 44307, Kaunas, Lithuania

Background: The majority of malignant tumors that occur within the central nervous system arise from supportive tissues know as the neuroglia. The neuroglia is composed of glial cells, several types of which can develop into tumors. The most common type of glial cell tumor is the astrocytoma, which can be classified according malignancy in four grades: from low grade glioma (LGG; grade I-II astrocytoma) to high grade (grade III astrocytoma, grade IV astrocytoma (glioblastoma)). In cancer, one of the epigenetic proceses - global alterations of DNA methylation play an important role in tumor initiation and progression. Hypermethylation mostly occurs at the promoter CpG Island of genes that are involved in processes leading to tumor formation and progression. Aberrant expression of NDRG2 (N-Myc downstream-regulated) gene could play a tumor suppressor role in carcinogenesis. NDRG2 seems to be more abundantly expressed in normal tissues, and is involved in the regulation of proliferation, apoptosis and differentiation. NDRG2 expression is reportedly decreased by promoter methylation in a variety of malignancies. TGFβ1 (transforming growth factor, beta 1) is a multifunctional cytokine that controls tissue homeostasis and embryonic development. TGFβ1 acts as a tumor suppressor in normal epithelial cells and earlystage tumors and becomes an oncogenic factor in advanced tumors. We investigate promoter methylation and protein expression of NDRG2 and TGFβ1in different grades of glioma. Material and Methods: The methylation specific polymerase chain reaction (MSP) was performed using bisulfite treated DNA. Proteins were separated according to their molecular weight by electrophoresis in 10 % (NDRG2) and 12 % (TGFβ1) polyacrylamide gels. Western blot was used to compare levels of protein expression between different grade gliomas. Results: The research of NDRG2 gene promoter methylation showed that methylation frequency depends on malignancy, which is different among brain tumor grades. Methylation frequencies of TGFβ1 gene were 22 % for astrocytoma I, 31 % for astrocytoma II, 10% for astrocytoma III and 11 % for glioblastoma. Slight decrease of TGFβ1 protein expression levels were observed in higher grade gliomas. In case of NDRG2 the protein expression decrease in higher grade gliomas is significantly more prominent. Conclusions: NDRG2 and TGFβ1 promoter methylation frequency depends on glioma grade. As the gliomas progress there is a reduction of TGFβ1 and NRDG2 expression in the cells, indicating that NDRG2 and TGFβ1 may play an inhibitory role during the development of gliomas.

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P-8 AREG PROMOTER METHYLATION IN DIFFERENT GRADES OF GLIOMA Rūta Sincevičiūtė 1.2, Paulina Vaitkienė 1 Laboratory of Neurooncology and Genetics, Institute of Neuroscience, Kaunas Academy of Medicine, Lithuanian University of Health Sciences, Eiveniu str. 4, Kaunas, LT 50009, Lithuania; The Faculty of Medicine, Lithuanian University of Health Sciences, Kaunas, Lithuania. A. Mickevičiaus str.9.

Abstract Background: Gliomas are a type of brain tumors that originates in the glial cells in the brain. AREG gene encodes a protein that is an autocrine growth factor as well as a mitogen for astrocytes, Schwann cells, and fibroblasts. This protein interacts with the EGF/TGF-alpha receptor to promote the growth of normal epithelial cells and inhibits the growth of certain aggressive carcinoma cell lines. The objective of the study was to determine the methylation frequency of AREG gene and to evaluate the associations between AREG promoter methylation and different grades of glioma. Method: 67 gliomas were analysed in the experiment: 8 were of the stage I, 10 – of the stage II, 8 of the stage III and 41 were of the stage IV. AREG gene promoter methylation was determined by methylation specific polymerase chain reaction (MS-PCR) after 33isulfate treatment. Also electrophoresis was performed. Results: The study of 67 gliomas showed that AREG gene promoter was methylated in 37 cases (55%) and was unmethylated in 30 cases (45%). 2 samples (25%), 4 samples (40%), 1 sample (12.5%), 30 samples (73%) of the tumors that were of the stage I, II, III and IV respectively were methylated whereas 6 cases (75%), 6 cases (60%), 7 cases (87.5%) and 11 cases (27%) of the tumors for stages I, II, III and IV respectively were unmethylated. Conclusions: Methylation of AREG gene in the promoter regions may play an important role in the development of gliomas. The methylation frequency is closely related to the malignancy of gliomas. Aknowledgement: This research was from the Research Council of Lithuania.

funded

by

a

grant

(No.

LIG-11/2012)

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P-9 TES GENE METHYLATION IN DIFFERENT MALIGNANCY GLIOMAS Indrė Valiulytė1.2, Paulina Vaitkienė 1 1

Laboratory of Neurooncology and Genetics, Institute of Neuroscience, Kaunas Academy of Medicine, Lithuanian University of Health Sciences, Eiveniu str. 4, Kaunas, LT 50009, Lithuania; 2 Department of Biochemistry and Biotechnologies, The Faculty of Natural Sciences, Vytautas Magnus University, Kaunas, Lithuania. Vileikos str. 8, LT-44404 Kaunas; [email protected]

The TES gene methylation is one of several potential epigenetic prognostic markers in glioblastoma. TES gene expression is involved in cell adhesion, cell spreading and in the reorganization of the actin cytoskeleton. Also, plays a role in the regulation of cell proliferation, acts as a tumor suppressor - inhibits tumor cell growth. Methods: Were investigated 95 patients with different grade gliomas: grade I - 11 samples, grade II - 25, grade III - 16 and grade IV (Glioblastoma) - 43 samples. Tumor DNA was extracted from 2540mg of frozen tissue and the methylation status of TES gene promoter was determined by bisulfite treatment of DNA. TES gene methylation was performed by using methylation specific polymerase chain reaction (MS-PCR) method. MS- PCR products were separated by electrophoresis on 2% agarose gels with ethidium bromide and visualized under UV illumination. Results: 61.05% of all patients had TES gene methylation. In different malignancy grade gliomas TES gene promoter methylation frequencies were varied. Glioblastomas demonstrated TES promoter methylation in 79.07% (34/43). Grade I (0/11) gliomas have not showed TES promoter methylation. TES promoter methylation was observed in 56% (14/25) of grade II gliomas and 62.5% (10/16) of grade III gliomas. In conclusion: TES gene methylation is related with glioma malignancy. TES gene methylation may be involved in gliomagenis process and might be one of the potential epigenetic prognostic markers in glioblastoma. Acknowledgement: This research was funded by a grant (No. LIG-11/2012) from the Research Council of Lithuania.

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P-10 COMMON MERGANSER (Mergus merganser) POPULATION ANALYSIS USING MITOCHONDRIAL DNA MARKERS IN LITHUANIA Auksė Abromaitytė1, Rūta Godliauskaite1, Aniolas Sruoga1,2, Vaida Tubelytė1, Vykintas Baublys1 1

Department of Biology, Vytautas Magnus University, Vileikos 8, Kaunas, LT 44404, Lithuania. E-mail address: [email protected] 2 Institute of Ecology, Nature Research Centre, Akademijos 2, LT 08412 Vilnius, Lithuania

Common merganser (Mergus merganser) is a member of Anatidae family and Merginae subfamily. Now they are found all over Lithuania country. Despite this fact species was very rare few decades ago in Lithuania and at this moment is written into The Red List of Lithuania fauna at 5(Rs) category. Till now, there are no researches done on the Common merganser population structure in Lithuania using molecular methods. Essential and significant aim of Common merganser studies in Europe [1, 2] is to explain and determine the flyways of the Common mergansers, since they are not clear. Because of the global climate changes, many of the migrating birds flying routes changes. It can make serious impact to endemic ecosystems. Also migrating birds are potential transfers of various pathogens. The aim of our research is supplement the knowledge of the European population of Common merganser structure and breeding habitats. To achieve this, we investigated mitochondrial DNA Dloop control region of migrating and breeding Common mergansers samples from Lithuania. The material for molecular research is collected in collaboration with the Institute of Ecology of Vilnius University. For molecular analysis samples were taken from 33 Common merganser blood and liver samples. The material was collected in 2003-2012 from the Baltic Sea coastal zone, Kaunas, and Vilnius regions. Polymerase chain reaction (PCR) was done, using mitochondrial DNA D-loop control region marker (MMCRL). The size of the DNA fragments was 514 nucleotides. Amplified PCR products were sequenced and analyzed by MEGA 5 software package. Sequences were compared with those obtained by other researchers by using the same MMCRL marker for samples from different European regions. The unique haplotypes were found in samples from Lithuania, that hasn‘t been yet detected in Common mergansers from other European regions. References 1. Gautschi B, Koller B (2005) Mol Ecol Notes 5:133–134 2. Hefti-Gautschi B., Pfunder M., Jenni L., Keller V., Ellegren (2009) Conserv Genet 10:87–99

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P-11 NUTRITIONAL AND NON-NUTRITIONAL ELEMENTS OF THE LEAVES OF HYMALAYAN BALSAM AND SMALL BALSAM SAMPLED FROM DIFFERENT LOCATIONS IN LITHUANIA Eglė Aleksaitė, Irma Šliumpaitė, Lina Zybartaitė, Rasa Jankauskaitė, Algimantas Paulauskas, Eugenija Kupčinskienė Department of Biology, Vytautas Magnus University, Kaunas, Lithuania, LT-44404

Data concerning plant element analyses might be helpfull tool in explanation and solving out problems related to alien plant invasions. Element concentrations reflect nutritional state of the plant, ability to accumulate elements and reflect quality of environment from which plants were taken. Element data may discover conditions the most favourable for the plants also might prompt ideas for the best eradication of the invasive species. Till now in literature described elemental analyses are of narrow scope in respect to the number of examined populations also very poor according to the number of elements investigated. Current study aimed at evalution of Lithuania populations of invasive Impatiens according to the leaf element concentrations also variations in soil ph with the purpose in perspective to compare physiological parameters with those obtained in molecular genetics analyses and habitat data. Hereby elemental analyses of small balsam (Impatiens parviflora) also Himalayan balsam (Impatiens glandulifera) were performed; also soil taken from splant sampling plots was evaluated according to pH. Nitrogen concentration was estimated by Kjeldahl method, concentrations of the other elements were defined using atomic absorption spectroscopy, and soil pH was estimated by pH-meter. Obtained results show that for Impatiens from more neutral soils (pH > 7), leaf potassium, calcium, magnesium and sodium concentration are much higher, and for Impatiens taken from more acidic soils (pH < 7), leaves have accumulated big amounts of iron, aluminium, zink, manganese and cupper. Our data demonstrantes that eutrophisation of environment creates favourable conditions of I. parviflora and I. glandulifera invasion. Concentrations of trace elements of Impatiens leaves did not exceed toxicity threshold defined for the same organs of the other systematically related species. Keywords: Kjeldahl method, AAS, soil, leaf, nutrients

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P-12 INFLUENCE OF CADMIUM IONS ON THE REDUCED GLUTATHIONE AND LIPID PEROXIDATION IN THE LIVER AND RED BLOOD CELLS OF MICE Kristina Čijauskaitė, Rasa Bernotienė, Leonid Ivanov Neurosciences Institute and Department of Biochemistry, Medical Academy, Lithuanian University of Health Sciences, Eiveniu 4, LT-50009, Kaunas, Lithuania

Abstract. Objective: The present investigation was undertaken to evaluate the influence of Cd on the reduced glutathione (GSH) and lipid peroxidation in liver and in red blood cells of mice. Methods: Experiments were done on outbred white laboratory mice using intraperitoneal injections of CdCl2 solution (14 μmol Cd/kg body mass). The exposure-time was 2 h, 8 h, 24 h and 14 days. GSH was measured by reaction with DTNB (5,5-dithiobis(2-nitrobenzoic acid)) to give a compound that absorbs 412 nm light wavelength. Lipid peroxides were estimated by measuring thiobarbituric-acidreactive substances and were expressed as MDA (malondialdehyde). Results: Our results indicate that at the 8th h the content of GSH in mice liver significantly increased by 35 %, meanwhile in red blood cells the content of GSH exceeded the control level at the 2nd h, 8th h and 24th h by 19 %, 35 % and 18 %, respectively. After 14 days of injections of CdCl2 solution, the content of GSH in liver and in red blood cells of mice decreased by 32 % and 29 % as compared to the control mice group, respectively. Further experiments carried out in order to examine influence of Cd ions on the content of MDA in liver and in red blood cells of mice after 2 h, 8 h, 24 h and 14 days of injections of CdCl2 solution. The results indicate that at the 8th h and 24th h, MDA content in mice liver was significantly increased by 236 % and 118 %, respectively. Meanwhile, MDA content in mice red blood cells in all time points were at the control level.

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P-13 MOLECULAR DETECTION AND CHARACTERIZATION OF ANAPLASMA PHAGOCYTOPHILUM STRAINS IN IXODIDAE TICKS AND RESERVOIR HOST Edita Juškaitė, Algimantas Paulauskas, Jana Radzijevskaja Irma Pūraitė Vytautas Magnus University, Vileikos 8, Kaunas, LT-44404, Lithuania

Abstract Anaplasma phagocytophilum is an obligate intracellular bacterium, which has a broad geographic distribution and the high degree of biological and clinical diversity. To investigate genetic diversity of A. phagocytophilum strains in Ixodidae ticks and a reservoir host were used three genes of pathogen genome were analyzed. Characterization of A. phagocytopilum strains were performed of partial 16S rRNA and major surface protein (msp4, msp2) genes. Nested PCR and sequence analysis were detected strains were compared with those originating from other European countries. In total were examined 66 pulls the ticks removed from migratory birds, 20 pulls the ticks from rodents, 53 spleens of rodents, 52 questing Ixodidae ticks in the areas of cervine animals, 41 spleens and 14 blood samples of cervine animals for infection. Two msp4 gene variants and a single 16S rRNA gene variant of A. phagocytophilum were found. The present study represents the first molecular characterization of A. phagocytophilum strains circulating in vectors and host of A. phagocytophilum in Lithuania. Keywords: Anaplasma phagocytophilum strains msp4 gene, msp2 gene, 16S rRNA gene

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P-14 DEVELOPMENT OF A MIXED-MODE SOLID PHASE EXTRACTION FOR A MULTI-RESIDUE ANALYSIS OF 15 BENZODIAZEPINES IN BLOOD BY RAPID GAS CHROMATOGRAPHY WITH NEGATIVE-ION CHEMICAL IONIZATION MASS SPECTROMETRY Nerijus Karlonas a, b, Arunas Ramanavicius c, Almira Ramanaviciene a a

Department of Analytical and Environmental Chemistry, VU, Naugarduko 24, LT-03225 Vilnius, Lithuania; E-mail: [email protected] b State forensic medicine service under the Ministry of Justice of the Republic of Lithuania, Didlaukio 86E, LT-08303 Vilnius, Lithuania; E-mail: [email protected] c Department of Physical Chemistry, VU, Naugarduko 24, LT-03225 Vilnius, Lithuania

Benzodiazepines constitute a large and important class of pharmaceutically active drugs with antiepileptic, hypnotic, tranquillizing, anticonvulsant, sedative, muscle relaxant, and amnesic properties [1]. Therefore, it is essential to develop a rapid and sensitive method for the determination of benzodiazepines. Gas chromatography with negative ion chemical ionization mass spectrometry (GC/NICIMS) is a sensitive and selective technique for the analysis of benzodia-zepines [2-4]. The mechanism of NICI is characterized as a “soft” ionization technique, whereby NICI spectra exhibit information about prominent molecular anions and therefore about their molecular weight. This method enables to reach the highest specificity for these analytes and meet the requirements of good laboratory practice. The proposed GC/NICI-MS method coupled with a mixed-mode solid phase extraction (a strong cation-exchange polymeric sorbent) and derivatization by N-(tert-butyldimethylsilyl)-Nmethyltrifluoroacetamide: acetonitrile: ethyl acetate mixture (20:40:40 (v/v/v)) was shown to be useful for the analysis of benzodiazepines and their metabolites in whole blood. Sample preparation conditions including selection of the solvent for washing and elution steps, pH values, the derivatization conditions, reaction temperature and duration were also optimized. To the best of our knowledge, this method has been used for the first time for the optimization of sample preparation at pH 1.0. Furthermore, the speed of the analytical separation was emphasized by modifying various GC/NICI-MS parameters. The developed method provides significant advantages in comparison with other previously published methods [1-5]. It shows higher sensitivity (the limit of detection ≤ 0.62 ng mL-1) in whole blood samples. Moreover, this method has several advantages: elimination of interferences, lowvolume of samples (0.2 mL), a multi-residue analysis, and very fast chromatographic separation of analytes (3.9 min). The reliability of this method was certified by means of an exhaustive validation study. Analysis performed in human whole blood samples showed that this method is sensitive, selective and specific enough to detect analytes after a long time of a single oral administration of some drugs. References 1. Samanidou VF, Uddin MN, Papadoyannis IN (2009) Bioanalysis 1:755-784. 2. Karlonas N, Padarauskas A, Ramanavicius A, Minkuviene Z, Ramanaviciene A (2012) Chemija 23:91-99. 3. Gunnar T, Ariniemi K, Lillsund P (2006) J. Mass Spectrometry 41:741-754. 4. Kaartama R, Jarho P, Savolainen J, Kokki H, Lehtonen M (2011) J. Chromatogr. B 879:16681676. 5. Karlonas N, Padarauskas A, Ramanavicius A, Ramanaviciene A (2013) J. Sep. Science DOI 10.1002/jssc.201201069.

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P-15 FLUORESCENCE OF ETHIDIUM BROMIDE IN SUSPENSIONS OF JURKAT AND MX-1 CELLS Mantas Riauka, Rimantas Daugelavičius Departament of biochemistry and biotechnologies, Vytautas Magnus university, Kaunas [email protected]

Abstract: We analyzed fluorescent properties of dsDNA intercalating agent ethidium bromide in suspensions of human leukemia Jurkat T-cells and human breast carcinoma MX-1 cells. TECAN Genios Pro 96/384 Multifunction Microplate Reader was used to assess the fluorescence. The obtained data indicate that intensity of ethidium fluorescence is rather low in suspensions of the intact cells. After permeabilization of the cells using digitonin, ethidium fluorescence substantially increased. These results suggest that the intracellular concentration of ethidium is diminished because of the efflux of this indicative cation from cells by multidrug resistance (MDR) pumps. Cell type-specific changes in fluorescence patterns were observed after lipophilic tetraphenylphosphonium ions-induced enhancement of the MDR pumps-encoding gene expression.

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P-16 PHYLOGENETIC ANALYSIS OF VIBURNUM (VIBURNUM) BASED ON MICROSATELLITE MARKERS S.Bogačiovienė1, J.Žukauskienė1*, A.Paulauskas1, Eugenija Kupčinskienė1, K.Senavaitytė1, L.Česonienė2, R.Daubaras2 1

Department of Biology, Vytautas Magnus University, Vileikos str.8, LT-44404 Kaunas Kaunas Botanical Garden of Vytautas Magnus University, Ž. E. Žilibero 6, LT-46324 Kaunas *Email: [email protected] 2

Viburnum (Viburnum) - is an old plant of our country, which is characterised valuable of nutritional and medical properties. The genus Viburnum includes more than 160 species of shrubs and small trees distrubuted widely througt out Northen hemispere into the Southern hemisphere (Winkworth and Donoghue, 2004). Viburnum species in ornamental plant wich provide many ideal year-round qualities (Dean et al., 2011). Over years molecular tools have become a superior method for hybrid analysis and cultivar identification or ornamental plants (Dean et al., 2011). In Vytautas Magnus University‘s Kaunas botanical garden is a rich collection of Viburnum, more than 20 Viburnum species. With this Viburnum gene collection there were no molecular research done until now. Thirty-three microsatellite markers were tested in our researh with 18 different species of Viburnum. For phylogenetic analysis 9 selected microsatellite markers were explored Viburnum collection from VMU Kaunas Botanical Garden. After analysis the most alleles were found with primer - VDO14. The most unique alleles were found with primer -VDO12. The primer VCC_I8 was as monomorfic, primer VCC_B3 were polimorfic, seven fragments were detected. Amplitude of allele‘s size was 82 to 292 (bp). Financial support for this study was provided by the Research Council of Lithuania by project “Change of bioactive compounds in underutilized berry species depending on genotype and environment” (No.SVE-04/2011). References:  D.Dean, X.Wang, W.E.Klingeman, B.H.Ownley. 2011 Screening and Characterization of 11 Novel Microsatellite Markers from Viburnum dilitatum. Hort. Sci. 46(11): 1456 – 1459.  Winkworth, R.C. and M.J Donoghue. 2004. Viburnum phylogeny: Evidence from the duplicated nuclear gene GBSSI. Mol.Phylogenet.Evol. 33: 109-126.

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P-17 GENETIC DIVERSITY APODEMUS SYLVATICUS AND APODEMUS FLAVICOLLIS IN LITHUANIA Karolis Sivickis, Algimantas Paulauskas, Vaclovas Gedminas Faculty of Natural Sciences, Vytautas Magnus University, Vileikos 8, LT- 44404 Kaunas, Lithuania,

Mice of genus Apodemus are wide spred in Europe, western Asia and North Africa. Twenty one species are now recognized, but morfological similarity makes difficulties to recognize these species. Therer is a requirement of molecular markers to clear species identification. The aim of our investigation was to evalute genetic diversity of A. sylvaticus and A. flavicollis in Lithuania using ISSR markers. For DNA research was captured 45 mices from Kaunas district, 13 from Zemaitija National park, 130 rodents from Neringa, 13 from Kaisiadoriai district, and 49 from Juodkrante. Also for research using other mouse from previos Vytautas Magnus University Natural science falculty expeditions from Norway, where were captured 15 Apodemus genus mouse. Using chloroform method „Genomic DNA Purification Kit“(Fermentas, K0512). DNA was extracted from ears and muscules for genome studies. Chosen primers: AP-1 (5‘-(CAG)5GC-3‘), AP-2 (5‘-(CAG)4AC-3’), AP-5 (5‘-(G)6GC-3’), E8S(5‘-TAAATGGGACAGGTAGGACC-3’) and Ep8 1 (5‘CCTTACTGCCTCTTGCTTC-3’). For PCR data analysis used horizontal 1,5% agarose gels. The size of the fragments ranged from 300 to 1700 bp. The primers AP-2, AP-5 and E8S were the most informative to study genetic diversyti of genus Apodemus. References: 1. J. R. Michaux, E. Magnanou, E. Paradis, C. Nieberding, R. Libois., Mitochondrial phylography of the woodmouse (Apodenys Sylvaticus) in The western Paleartic region. Mol Ecol 12:685-697. 2. Bugarski-Stanojevic V. J. Blagojevic, T. Adnadevic, V. Jojic and M. Vujosevic, Molecylar phylogeny and distribution of three Apodemus species (Muridae, Rodentia) in Serbia, Zool Syst Evol Res (2008) 46(3), 278-286 3. Jelena Blagojevic, Gorana Stamenkovic, Tanja Adnadevic, Eva B. Giagia-Athnasopoulou, Mladen Vujosevi,. Comparative study of the phylogenetics structure in six Apodemus (Mammalia, Rodentia) inferred from ISSR-PCR data, Systematics and Biodiversity, 2011. 90:1, 95-106.

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P-18 GENETIC DIVERSITY OF IMPATIENS GLANDULIFERA IN SOME NORTHERN EUROPE SITES ESTIMATED BY ISSR MARKERS Kristė Stravinskaitė, Lina Zybartaitė, Judita Žukauskienė, Algimantas Paulauskas, Eugenija Kupčinskienė* Department of Biology, Vytautas Magnus University, Vileikos 8, LT-44404 Kaunas, Lithuania * Corresponding author. E-mail: [email protected]

Himalayan balsam (Impatiens glandulifera) is an invasive alien annual up to 3 m height with pink or purple flowers plant [1]. Native to Himalayan region, I. glandulifera nowadays is among the most invasive plants in almost all Europe. Many introductions of the species are the result of their usage as decorative plant [2]. Till now little has been known about the genetic diversity of I. glandulifera in the northern parts of Europe. The objective of this study was to evaluate genetic diversity of I. glandulifera growing in Baltic countries (Lithuania, Latvia, Estonia) using Inter Simple Sequence Repeat (ISSR) markers. A total of 11 I. glandulifera populations were sampled. The plants were collected from 4 Lithuania, 4 Latvia and 3 Estonia populations (15 individuals in each population). Four ISSR markers (ISSR2, ISSR3, ISSR4, ISSR5) were used for analyses [3]. Size of the DNA fragments in Lithuania, Latvia and Estonia was similar, ranging from 190 to 1300 bp. The highest number of fragments was estimated in Lithuania (18–21), less number of the fragments was obtained in Latvia plants (10–21) and the least – in Estonia (10–15). The UPGMA dendrogram revealed clear differentiation between the populations. ISSR analyses indicate that highest I. glandulifera genetic diversity was found in Lithuania as the most southern location country. Keywords: Himalayan balsam, invasion, alien plant, population genetics, molecular markers References 1. 2. 3.

Clements DR, Feenstra KR, Jones K, Staniforth R (2007) The Biology of Invasive Alien Plants in Canada. 9. Impatiens glandulifera Royle. Canadian Journal of Plant Science, 403–417. Beerling DJ, Perrins JM (1993) Impatiens glandulifera Royle (Impatiens Roylei Walp.). Biological flora of British Isles, No. 177. Journal of Ecology 81 (2): 367–382. Provan J, Love HM, Maggs CA (2007) Developments of microsatellites for the invasive riparian plant Impatiens glandulifera (Himalayan balsam) using inter simple sequence repeat cloning. Mol Ecol Notes 7: 451–453.

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P-19 ANALYSIS OF GENETIC VARIABILITY OF BLUEBERRIES (VACCINIUM L.) CULTIVARS USING MICROSATELLITE MARKERS S.Bogačiovienė1, J.Žukauskienė1*, A.Paulauskas1, Eugenija Kupčinskienė1, I.Šarauskienė1, L.Česonienė2, R.Daubaras2 1

Department of Biology, Vytautas Magnus University, Vileikos str.8, LT-44404 Kaunas Kaunas Botanical Garden of Vytautas Magnus University, Ž. E. Žilibero 6, LT-46324 Kaunas *Email: [email protected] 2

Blueberries (Vaccinium corymbosum L.) - one of the newest garden plants. Blueberries were started cultivate in the beginning of twentieth century in the United States. Berries have many biological compuonds such as anthocyanins, polyphenol and flavonoids. These compounds have anticarcinogenics characteristics, they protect cells against free radicals, slows the aging process. For our genetic analysis we were used young blueberry leaves, which were collected in Vytautas Magnus University Kaunas Botanical Garden at May 2011 and in Vilnius University Botanical Garden at September 2011. The DNA was extracted of young blueberry leaves with a DNA-silica column. For blueberries research we used 15 microsatellite primers designed by Boches et al. (2005). Then we visualized fragments in a vertical poliacrilamid gel electrophoresis. Fifteen primers were checked for blueberries, after that analysis 8 microsatellite primers were selected: CA23F, NA961, NA1040, CA421F, CA169F, CA112F, VCC_J9, VCC_S10. The total number of fragments was ranged from 4 to 29, and the size - from 115 to 250 bp. The most unique alleles were detected in 'Chanticleer' and 'Sila' species, at least - one unique allele was found in cultivars: 'Aron', 'Atlantic', 'Bluetta', 'Earliblue', 'Croatan' and 'Blueheaven'. References: Boches P. S., Bassil N. V., Rowland L. J. 2005. Microsatellite markers for Vaccinium from EST and genomic libraries. Molec. Ecol. Notes 5: 657–660. Keywords: blueberry, vaccinium, microsatellite markers.

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P-20 RESEARCH ON TERRESTRIAL TESTATE AMOEBA (TESTACEA) IN LITHUANIA Šatkauskienė Ingrida, Štuopytė Gertrūda, Šidlauskaitė Lina, Kantautė Vaiva, Vytautas Magnus University

Abstract The testate amoebae are a group of heterotrophic protists which is mostly found in freshwater, seawater, soil, moss and lichens. These amoebas feed on bacteria, algae and other protists so they are important for normal function of water and land ecosystems. However the fauna of terrestrial testate amoebas of Lithuania remains poorly know. During 2009-2010, terrestrial testate amoebas were studied in mosses (Bryophyta) which were collected from different places of Lithuania. It was found out, that diversity of testate amoeba was higher in moss collected in clean areas rather than in potentially polluted areas (moss were sampled from the areas near highways). In this study there was found 28 species, which belong to 12 genera of testate amoebas.

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P-21 THE EFFECTS OF LEAD AND NICKEL IONS ON TOTAL PROTEINS AND METALLOTHIONEINS SYNTHESIS IN MICE LIVER Indrė Šveikauskaitė, Jurgita Šulinskienė, Ilona Sadauskienė, Leonid Ivanov Neuroscience Institute, Medical Academy, Lithuanian University of Health Sciences, Lithuania E-mail: [email protected]

Abstract The present study was conducted to investigate the effects of lead and nickel ions on total proteins and metallothioneins synthesis in mice liver. Introduction: Lead (Pb) is a heavy metal used in a wide variety of consumer products and occupational settings [1]. Nickel (Ni) is an ubiquitous metal element found in a wide variety of compounds [1]. The protein synthesis is the process by which biological cells generate new proteins. Protein synthesis is divided into three stages: initiation, elongation and termination [2]. Metallothioneins (Mts) are a group of small proteins, found in the cytosol of cells, particularly of liver, kidney, and intestine [3]. Materials and methods: Experiments were done on 4-6 weeks old white laboratory outbreed mice weighing 20-25 g. Concentration of protein was determined by Lowry method. MTs were assayed in mice liver according to the method of Peixoto N. C. Results: There is no statistically significant effect of lead on protein synthesis in mice liver. According to the data, injections of NiCl2 solution, marked amino acid actuation to new synthesized proteins has increased. Obtained data showed, that in mice liver treated with NiCl2 and Pb(CH3COO)2 solutions, MTs content was increased. References [1] Richard C. Dart, M.D., PH.D. (2000) The 5 minute Toxicology consult. A Wolters Kluwer Company. [2] Michael Lieberman, Allan D. Marks, Colleen Smith (2007) Essentials of medical biochemistry. A clinical approach. Lippincott Williams & Wilkins [3] David A. Bender, Kathleen M. Botham, Peter J. Kennelly, Victor W. Rodwell, P. Anthony Weil (2009) Harper’s illustrated biochemistry. The McGraw – Hill Companies

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P-22 INVESTIGATION OF NITROGEN CONCENTRATION IN HYPERICUM MACULATUM CRANTZ POPULATIONS IN SOME BALTIC SITES Indrė Užubalytė, Irma Šliumpaitė, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė Department of Biology, Vytautas Magnus University, Kaunas, Lithuania, LT-44404

Hypericum species are widely used as traditional medicinal plants also in pharmaceutical industry. Despite the fact that Hypericum genus is one of the most important medicinal plants its nutritional aspects are not discussed in wider scope at both international and national level. Hypericum habitats vary in high extent, which is why it might grow in soils containing different amounts of nitrogen. However the consequences of different availability of main nutrition elements are not widly documented, especially in terms of partitioning of nitrogen between various aboveground organs. Hypericum maculatum is the second species according to abundance of Hypericum genus in Lithuania. The main purpose of this investigation was to measure the concentration of nitrogen in various aboveground organs (flowers, stems and leaves) of Hypericum maculatum. The material was collected in 12 populations from different areas in Lithuania: Lyduvėnai, Paltupiai, Katauskiai, Kretinga, Padubysys, Gruzdiškė, Girkalnis, Daugodai, Šilalė, Laveniškės. Dry material was grounded to a powder and nitrogen concentration was determined by Kjeldhal method. Nitrogen concentration in the flowers ranged between 1.6–2.5 % d. m. Nitrogen concentration of Hypericum maculatum collected in different populations varied in the range 1.8–2.7 % d. m. for the leaves and 0.5–0.9 % d. m. for the stems. The amount of nitrogen concentration fell into following order stems < flowers < leaves. In comparison to other plants, the concentration of nitrogen of Hypericum maculatum is at intermediate level. The recorded data are important for understanding possibilities how to grow Hypericum maculatum for medicine purposes. Key words: Kjeldahl method, nutrition, Malpighiales, Hypericaceae, leaf, stem, flower.

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P-23 GENETIC ANALYSIS OF IMPATIENS GLANDULIFERA POPULATIONS USING ISSR MARKERS Justina Vaitkevičiūtė, Lina Zybartaitė, prof. Eugenija Kupčinskienė, prof. Algimantas Paulauskas Vytautas Magnus University, Department of Biology, Vileikos 8, LT-44404 Kaunas, Lithuania.

Impatiens glandulifera is annual weed that came from West of Himalays and nowadays is highly invasive in almost whole Europe and occurs in various habitats. As a result of this fact Impatiens glandulifera displace native species and occupies growing areas near the rivers. Population genetic data on genetic peculiarities may aid in the management of the invasive species. We collected Impatiens glandulifera plants from 13 different abitats located in Lithuania and in the other European countries: Holland, Belgium, Luxembourg, Germany (Heepen, Rinteln, Bad Zalzuflen, Seeker, Switzerland, Austria, Hungary and Czech Republic. These populations we compared using an intersimple sequence repeat (ISSR) markers: ISSR2, ISSR3, ISSR4 and ISSR5. PCR products were separated by 1.5% agarose gel, photographed and statistically analyzed. Fragment length was measured by Gene Ruller (100 bp length standards). Among populations of Impatiens glandulifera genetic parameters ranged in the following intervals: percentage of polymorphic DNA within populations was between 5.6% (Switzerland and Lithuania) and 30.3 % (Germany, Bad Zalzuflen), mean being – 16.1%. In accordance with Nei’s gene diversity index, the most different were northern Luxembourg and Lithuanian populations (0.570), and the most simillar were two populations from Germany (Bad Zalzuflen and Rinteln populations; 0.831). It was found genetic difference between populations – 57%, within population 19% and 24% of the molecular diversity within countries. The principle coordinate analyses of populations and genetic tree built on the bases of UPGMA cluster analyses showed relations to geographical position of sampling sites only for part of populations. It might be explained by the fact that Himalayan balsam is spreading not only naturally - along streams, ditches, but it is unintentionally and intentionally distributed by the human in different distances and directions.

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P-24 SCREENING OF EPIPHYTIC PINE BACTERIA FOR ANTAGONISTIC PROPERTIES Viktorija Vaštakaitė, Odeta Buzaitė Vytautas Magnus University, Department of Biochemistry and Biotechnologies, Vileikos 8, LT-44404 Kaunas, Lithuania

Plant-associated bacteria can affect plants in different ways, including plant growth promoting. Beneficial effect on plant growth can be conditioned directly, by providing plants with nutrients nitrogen, iron or soluble phosphate or by producing plant hormones by bacteria. Indirect plant growth promotion can be conditioned by inhibition of growth of pathogenic microorganisms. In this study we investigated bacteria from surface of pine needles and screened 15 isolates for antagonistic properties which can be beneficial to plants. Ability of bacterial isolates to secrete antifungal compounds was tested. Studies were conducted to investigate the effect of epiphytic bacteria on growth of the Bacillus subtilis, Micrococcus luteus, Escerichia coli, Pseudomonas aeruginosa, Salmonella typhimurium (SL 1344), Staphylococcus aureus bacteria cultures. Epiphytic bacteria cross-interaction studies were also carried out by agar diffusion method. Surface of plant leaf - phylloplane along to other epiphytic bacterial habitats is promising source of microorganisms with plant growth affecting abilities including those with biotechnological potential.

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P-25 GENETIC DIVERSITY OF LITHUANIA POPULATIONS OF COMMON JUNIPER (JUNIPERUS COMMUNIS L.) EVALUATION ACCORDING TO ISSR MARKERS Alma Vilkaitytė, Ramūnas Vilčinskas, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė* Vytautas Magnus University, Department of Biology, Vileikos 8, LT-44404 Kaunas, Lithuania. e-mail: [email protected]

Common juniper (Juniperus communis L.) is the only Juniperus species that occurs in both hemispheres (Farjon, 1998). In Lithuania, population of common juniper covers an entire country and mostly concentrated in the pine forests of the South Lithuania (Navasaitis, 2004). Data concerning genetic diversity is an important source of information for both fundamental (through Lithuania and other countries of juniper populations viability and other biological properties), and practical applications in breeding and plantation work. Till now studies about genetic diversity of Lithuania population of Juniperus communis are missing. We have selected ISSR markers former applied to Central Europe populations of this species by Adams et al. (2003). DNA was extracted by modified CTAB method (Doyle and Doyle, 1990). We screened 15 oligonucleotides primers using ISSR–PCR, and reproducible amplification patterns were obtained with seven ISSR primers: 807 – AGAGAGAGAGAGAGAGT; 808 –AGAGAGAGAGAGAGAGC; 823 – TCTCTCTCTCTCTCTCC; 836 – AGAGAGAGAGAG AGA GYA ; 846 – CACACACACACACACART; 847 – CACACACACACACACARC and 857 – ACACACACACACACACYG. Totally, 14 populations with 10 individuals in each were analysed for polymorphic ISSR loci. The analyses revealed that the number of different lenght DNA fragments per population fore each loci varied from 5 to 14, with a size range of 180–1400 bp. Investigated ISSR loci revealed high polymorphism of populations of Juniperus communis. Keywords: conifers, Juniperus communis, ISSR markers, CTAB method, molecular markers, genetic diversity. References 1. Adams R.P., Pandey R.N. 2003. The concordance of terpenoid, ISSR and RAPD markers, and ITS sequence data sets among genotypes: an example from Juniperus. Biochemical Systematics and Ecology, 31, 375–387. 2. Doyle J.J. and Doyle J.L. 1990. Isolation of plants DNA from fresh tissue. Focus, 12 (1), 13–15. 3. Farjon, A. 1998. World Checklist and Bibliography of Conifers. Royal Botanic Gardens, Kew, UK, 298 – 299. 4. Navasaitis M. 2004. Dendrologija. Margi raštai, Vilnius, 244 – 248.

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P-26 HYPOCHLOROUS ACID CREATES PORES IN THE PLASMA MEMBRANE OF HUMAN ERYTHROCYTES WITH THE SIZE CLOSE TO THE SIZE OF ELECTRICAL PULSE-GENERATED PORES J. Vismantaitė, R. Rodaitė–Riševičienė, G. Saulis Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, Kaunas, Lithuania

Cell electroporation is widely used in cell biology, biotechnology and medicine. However, when a high–voltage is applied to the electrolyte solution, besides a cell membrane permeabilization, a variety of electrolysis reactions occur at the electrode–solution interfaces. During anodic processes, Cl2 gas is formed at the anode. Chlorine ions react with the water molecules and form hypochlorous acid – a very strong oxidizer, which changes chemical composition or pH of the experimental medium and can cause the cell death. This may have an influence considering electroporation effectiveness results. Studying electrochemical processes, which occur during high-voltage pulses and their consequences, is necessary, as they are important for optimizing practical applications. In this study, the membrane and cellular events caused by the action of hypochlorous acid (HOCl) on human erythrocytes – increase of the permeability of the cell plasma membrane and subsequent haemolysis of erythrocytes – were studied. The HOCl induced haemolysis was mediated by pore formation in the cell plasma membrane. The estimated pore radius was approximately 0.3–0.5 nm, which is close to the size of electrical pulse-generated pores. The results of this work can be useful for optimizing the electroporation methods used in biotechnology, medicine, and food industry.

Keywords: cell electroporation, electrochemical reactions, hypochlorous acid.

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P-27 PHYTOTOXICITY OF AMIDOSULFURON (SULFONYLUREAS HERBICIDE) TO AQUATIC MACROPHYTE LEMNA MINOR Jūratė Žaltauskaitė 1, Roberta Norvilaitė 2 1, 2

Vytautas Magnus university, Vileikos st. 8-223, LT-44404 Kaunas, Lithuania. E-mails: [email protected]; [email protected]

Abstract. This study examines effects of sulfonylurea herbicide Sekator OD 375 (amidosulfuron and iodosulfuron) to non-target aquatic plant species common duckweed (Lemna minor L.). The plants were affected by 5-75 µl/l concentrations of herbicide in their growth medium. L. minor had a strong phytotoxicty response to herbicide application (inhibition of growth, biomass; reduced content of photosynthetic pigments and increased content of malondialdehyde). The growth rate of L. minor was inhibited by 54.17 %. The dry biomass was by 57.2 % lower than in control. Application of herbicide provoked an oxidative stress, the concentrations of MDA duckweed was by 83.32 % higher than in control. Keywords: herbicides, non-target aquatic plant; oxidative stress, phytotoxicity

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P-28 RAPID ANALYSIS OF GENETIC DIVERSITY AMONG LITHUANIA POPULATION OF IMPATIENS NOLI TANGERE L. Julija Petkevičiūtė, Lina Zybartaite, Ramūnas Vilčinskas, Algimantas Paulauskas, Eugenija Kupcinskiene Vytautas Magnus University, Department of Biology, Vileikos 8, LT-44404 Kaunas, Lithuania

Balsaminaceae family comprises more than 1000 species, among them five species are present in Europe. There are 4 species in Lithuania, one of them is cultivated – Impatiens balsamina L., two species are known as aliens – Impatiens parviflora DC., Impatiens glandulifera L. and only one is growing naturally – Impatiens noli-tangere L. Valuable information might be obtained comparing adaptation level and variability of alien species and native ones. In Lithuania, molecular studies have been carried out only for invasive species of Impatiens, so the objective of this study was to evaluate the genetic variability of populations of Impatiens noli-tangere differing in geography or habitats by the randomly amplified polymorphic DNA (RAPD). Plants were collected from – Kaunas-Grabučiškiai, Kaunas-Marvelė, Kaunas-Raudondvaris, Kaunas-Romainiai, VarėnaPuvočiai, Seredžius, Rusnė, Juodkrantė, Palanga-Plazė, Plateliai, Kelmė-Šventragis, Kryžkalnis, Rokiškis-Panemunėlis, Jonava-Upininkai, Anykščiai. For I. noli-tangere studies eight different oligonucleotide primers (OPA-20, OPD-20, 222, 250, 269, 340, 474, 516) were used. Among populations of I. noli-tangere genetic parameters ranged in the following intervals: 3–15 % of polymorphic DNA bands, 0.005–0.049 for Nei’s gene diversity, 0.009–0.074 for Shannon’s information index. Pairwise genetic distances between populations ranged in the interval 0.185– 0.467. Genetic distance-based cluster analyses for 220 individuals indicated 7 major groups of populations (A – Kaunas-Grabučiškiai; B – Palanga-Plazė, Kryžkalnis, Kaunas-Raudondvaris; C – Anykščiai, Kelmė-Šventragis, Seredžius, Kaunas-Marvelė; D – Rokiškis-Panemunėlis, Rusnė; E – Juodkrantė, Plateliai; F – Kaunas-Raudondvaris; G – Jonava-Upininkai, Varėna-Puvočiai) among which there was no clear geographical pattern. On the bases of RAPD markers prepared UPGMA dendrogram of individuals and populations did not show tight relation between population geography and geographical distances. The same was true when principal component analyses were done for genetical data.

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P-29 DIVERSITY AND SEASONAL VARIATIONS IN PERIPHYTON COMPOSITION IN FRESHWATER ECOSYSTEMS Rasa Glasaitė, Ingrida Šatkauskienė Vytauto Didžiojo universitetas

Periphyton is a community of organisms, such as microscopic protozoan, algae, that live attached to stones, plants or other submerged surfaces in the water. Periphyton are primary producers and an important foundation of many water ecosystems food webs. These organisms also stabilize substrates and serve as habitat for many other organisms. Periphytonic organisms are used as biological indicators, because they are sensitive to water quality. The aim of this work was to try to establish composition and diversity of periphyton in stagnant and flow freshwater systems and evaluate the ecological quality of these ecosystems on the basis indication of periphyton. Periphyton was investigated in Nemunas river and in Kaunas lagoon. Periphyton samples were collected from the stones at regular intervals to compare periphytonic community structure and to examine its temporal changes. The results showed differences in the periphyton structure depending on the month (season) and on the type of water system. Totally, were collected and identified 15 species and 13 genus of periphyton. The dominant genus in all locations was Euglena sp. and diatoms Navicula lanceolata, Craticula cuspidata (Navicula cuspidata), Amphora ovale. All these organisms are good water contamination indicators. Navicula sp. and Euglena sp. were found more abundance in Kaunas lagoon than in Nemunas river, so this preliminary results shows that lagoon is a higher pollution level.

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P-30 LOW UV-B RADIATION AND SUBSTRATE ACIDITY IMPACT ON PHOTOSYNTHEIC PARAMETERS OF BRASSICA NAPUS Irena Januškaitienė Vytautas Magnus University

The aim of this study was to assess combined effect of 1 kJm-2d-1 UV-B radiation and substrate acidity (pH 4.8) on photosynthesis and growth of summer rape (Brassica napus) in a controlled environment. Rape were sown in a neutral (pH 6.0–6.5) peat substrate, when 2nd true leaf unfolded, growth substrate was acidified using 0.07 mM H2SO4 concentration solution and UV-B radiation exposure was started. Duration of the experiment: five days. Gas exchange parameters were measured with portable photosynthesis system LI-6400. Content of chlorophyll was evaluated spectrophotometrically. The results showed that the highest inhibition on Brassica napus growth and photosynthetic parameters was under combined effect of 1 kJm-2d-1 UV-B radiation and pH 4.8 of substrate acidity. Single 1 kJm-2d-1 UV-B radiation impact on investigated parameters was low and statistically insignificant, except transpiration rate and water use efficiency. Single substrate acidity affected in 58.6 % (p < 0.05) decrease in photosynthetic rate, almost twofold decrease of transpiration rate, and 3.8 % (p < 0.05) decrease in intercellular CO2 concentration, and 27.1 % (p < 0.05) increase in water use efficiency, compare to the reference treatment. Combined substrate acidity and UV-B radiation effect on photosynthetic rate was the highest, when the decrease was 80.6 % (p 0.05). And the increase (by 2 %, p < 0.05) of intercellular CO2 concentration of Brassica napus affected by combined impact of investigated stressors shows that there higher negative impact was on enzymatic reactions of photosynthesis than on transpiration. The highest negative effect on content of photosynthetic pigments and biomass accumulation also was of combined effect of investigated stressors.

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P-31 MEASUREMENT OF NITROGEN DIOXIDE CONCENTRATION IN COLD AND WARM SEASONS USING A PASSIVE SAMPLING METHOD Dovilė Laurinavičienė, Audrius Dėdelė Vytautas Magnus University, Department of Environmental Sciences

Abstract The aim of the present study was to assess distribution of nitrogen dioxide concentration in Kaunas city during cold and warm seasons. A passive sampling method was used. Nitrogen dioxide concentration was measured at 39 sampling sites in Kaunas city. Study was performed in 2010– 2011. The study results showed that mean of nitrogen dioxide concentration in Kaunas city was 12.0 µg/m3. Nitrogen dioxide concentration in cold season (winter) was 18.2 µg/m3, in warm (summer) 5.7 µg/m3. The highest nitrogen dioxide concentration was obtained near street site (14.9 µg/m3), the lowest – near regional background site (9.4 µg/m3) and near urban background site it was 11.4 µg/m3. The results of the study were introduced in the ESRI Geographic Information System software ArcGIS and the extension Geostatistical Analyst was used to obtain maps of distribution of nitrogen dioxide concentration. Lowest nitrogen dioxide pollution area was in districts which are located in the periphery of the city. The highest concentration of nitrogen dioxide was in Centras, Žaliakalnis, Šančiai, Kalniečiai, Sargėnai. It is caused by intensive traffic and unfavorable geographical position. In these districts there are more busy roads and more buildings. Centras district is located in a valley; there are unfavourable conditions for pollution dispersion. Keywords: air pollution, nitrogen dioxide concentration, cold and warm seasons, traffic intensity sites types, interpolation method

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P-32 SOIL NET CO2 EXCHANGE RATE RESPONSE TO DIFFERENT TILLAGE AND FERTILIZATION SYSTEMS Agnė Putramentaitė1, Mantas Butrimavičius2, Simona Pranaitienė3 1,3

Institute of Agriculture, Lithuanian Research Centre for Agriculture and Forestry; 2Vilnius University; E-mail: [email protected]; [email protected]; [email protected]

Abstract It is no consensus worldwide concerning soil management practices on CO2 emissions. The main goal of the study was to determine soil CO2 emission in different agricultural management systems under contrasting meteorological conditions. Two-factorial field experiment was set up at the Institute of Agriculture, Lithuanian Research Centre for Agriculture and Forestry on an Endocalcari-Epihypogleyic Cambisol. Tillage systems – conventional tillage (CT) and no-tillage (NT) were the main plots, while fertilization – not fertilized (1) and moderate rates of mineral NPK fertilizers (2) were as sub-plots. Data revealed that soil net CO2 exchange rate (NCER), temperature (T-soil) and volumetric water content (W) significantly responded to meteorological conditions of individual year (2009, 2010, 2012) and soil management practices. Three year study revealed that rainy weather conditions suppressed NCER sharply. Soil temperature and water content in both unfertilized tillage systems (CT-1, NT-1) were the main factors influencing CO2 emission. Fertilization with mineral NPK fertilizers increased CO2 emissions on average by 32% compared to unfertilized plots, but did not eliminate suppressive effects of meteorological conditions on NCER. CO2 emission under moderate climatic condition in Lithuania was about 4-5 folds lower than in the countries with tropical and subtropical climatic conditions. Keywords: soil, CO2 flux, tillage, fertilization, weather conditions

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P-33 DISEASES OF MEDICINAL PLANTS IN KAUNAS BOTANICAL GARDEN OF VYTAUTAS MAGNUS UNIVERSITY IN SPRING SEASONS Vilija Snieškienė, Antanina Stankevičienė, Ona Ragažinskienė Vytauto Didžiojo universiteto Kauno botanikos sodas, Ž. E. Žilibero 6, 46324 Kaunas, [email protected]

In 2006–2012 at spring seasons the state of plants of 350 species were assessed. Fungi of 10 species and 8 genera, grown in Kaunas Botanical Garden, causing medicinal plant fungal diseases occurring in spring were detected. Some fungal diseases are also noticed in early spring just after snowmelt. That is leave spot which occur at previous year and remain on over-wintered leaves. Such diseases are noticed annually on Oenothera biennis, Primula veris, Hedera helix and Bergenia crassifolia, disease agents are fungi belonging to Boeremia, Septoria, Ramularia, Cercospora genera. Hepatica nobilis leave spot, agent Septoria hepaticae, occurs before the ending of Hepatica blossom and also after the blossom, on the majority of leaves appear medium-sized irregular spots. Following group of diseases observed in spring is leave rust. Alium genus plants may be host plants for several genus of rust. A. ursinum is one of the host plants for ransons rust (Puccinia sessilis). Aecia (the first stage of fungus development) has a bright orange color, about 1-2 cm in diameter, is on the under side of the leaf of ransons. Later fungus development stages (uredium and telium) continue proceeding on Tyhoides arundinacea (L.) Moench. Puccinia sessilis can injure other medicinal plants (Convalaria majalis L.) and Paris quadrifolia L.), still rust on these plants was not found in Kaunas Botanical Garden. Allium giganteum is injured by Melampsora genera rust. Fungus Puccinia violae effects many species of viola. The fungus passes all stages of development on a single host plant – pansy viola. In spring on the under side of a leaf forms aecia concentrations, in such places leaf blade is slightly deformed (depressed), injured leaves look blistered. Since 2010 on young leaves of Sanguisorba minor aecia are observed. The part of young leaves and stalks are covered with bright orange color aecia, which drop out spores easily. All plants were damaged by leave rust, though the damage was of a small grade. The disease is not yet very harmful, as later grow new healthy leaves with no rust. Our climatic conditions in spring seasons are favorable to the spread of rot causing fungi, particularly to Botrytis genus. B. paeoniae – rot agent, to which Paeonia anomala and P. officinalis are susceptible. Young sprouts start showing black up from the root crown, cease growing, their peaks turn red. In wet weather injured parts may encrust with grey plaque – fungi conidium with easily dusting away conidia. Over the past few years in Lithuania spread Phytophthora genus pathogens. They are the fungi-like organisms. Vinca are vulnerable to them. In 2008-2009 part of V. minor has died and remained ones are currently healthy. The disease symptoms: old leaves turn brown from the base. Plants die in separate plots. Injury continues spreading from roots through water dishes. Most fungal pathogens are most active in summer; consequently diseases on all plants are more rare in spring seasons. Only 12 species, i.e. only 3.4%, out of 350 species of medicinal and spice plants grown in Kaunas Botanical Garden with the symptoms of fungal diseases were detected in spring time. Furthermore, these mostly are specific diseases typical only to this season, and require investigation as well. Leave spots, which remain on last-year leaves of the biennial or perennial plants, mature spores and continue spreading. The disease is not so harmful to the collections and exhibitions, but in industrial crops can make a significant loss. For instance, leaf blotch is a common disease for cowslip primrose and in spring it causes death for many industrial plants.

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P-34 BIOMONITORING OF URBAN AIR QUALITY IN KAUNAS, LITHUANIA, USING TRANSPLANTED LICHENS Gintarė Sujetovienė Department of Environmental Sciences, Vytautas Magnus University, Vileikos st.8, LT-44404 Kaunas, Lithuania E-mail: [email protected]

Abstract. The aim of the study was to evaluate air quality using an active biomonitoring method. The lichens Ramalina farinacea and Evernia prunastri were transplanted to urban sites in the city of Kaunas, Lithuania. Changes in chlorophyll a, chlorophyll b, phaeophytization ratio, electrical conductivity were determined after one, two and three months. Despite of increase in photosynthetic pigments concentration one month later further it content decreased in urban environment. No significant variation in content of chlorophyll were observed in comparison with the control and this could be attributed to variables of air pollution did not exceed threshold levels. There was no interspecific difference in the response of the two lichen species in the physiological parameters but Ramalina farinecea was more sensitive to cell membrane damage than Evernia prunastri in the studied urban environment. The study provides information that one month of transplantation is not enough in order to determine air quality status in urban environment. Keywords: lichen transplants, urban environment, air pollution

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P-35 VARIATION OF FLAVONOIDS AND PHENOLIC ACIDS IN FRUITS OF GENUS SORBUS L. PLANTS Kristina Gaivelytė, Valdas Jakštas, Valdimaras Janulis Lithuanian University of Health Sciences, Medical Academy, Faculty of Pharmacy, Department of Pharmacognosy. A. Mickevičiaus 9, Kaunas LT-44307, Lithuania e-mail [email protected]

Rowan (Sorbus L.) genus (Rosaceae) is widespread in the world. Various rowan species are used for food and as a source of medicinal raw plant material. Our research aim was to investigate quantitative composition of neochlorogenic acid, chlorogenic acid, rutin, hyperoside and isoquercitrin in fruits of the introduced rowan plants using HPLC method. In fruit samples quantitative estimation of neochlorogenic acid (0.932-5.435 mg/g), chlorogenic acid (0.553-7.496 mg/g), rutin (0.020-0.393 mg/g), hyperoside (0.016-1.190 mg/g) and isoquercitrin (0.018-0.655 mg/g) was performed. Total content of neochlorogenic acid, chlorogenic acid and identified flavonoids varied from 2.245 to 11.191 mg/g. Comparison of raw material samples of Sorbus species according to quantitative composition of identified phenolic compounds using cluster analysis singularized S. commixta Hedl. fruit samples. The samples comprised the highest contents of chlorogenic acid, hyperoside and isoquercitrin, but also the lowest contents of rutin. The highest contents of neochlorogenic acid were recorded in S. lancifolia Hedl. fruits, and those of rutin – in S. anglica Hedl. fruits. Fruit raw material of these plants could be the source of raw plant material rich in these compounds, promising aquiring phytotherapeutic agents with specific effects. Keywords: Sorbus, HPLC, flavonoids, phenolic acids.

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P-36 NATURAL VEGETABLE OILS POTENT ANTIOXIDANTS FOR NATURAL COSMETIC Giedrė Kasparavičienė, Dovilė Bajoriūnaitė, Saulė Velžienė, Zenona Kalvėnienė Department of Drugs Technology and Social Pharmacy, Faculty of Pharmacy, Medical Academy, Lithuanian University of Health Science A. Mickevičiaus 9, LT – 44307 Kaunas, Lithuania

Natural cosmetics have become a major trend in recent years. There is a growing awareness that we are responsible for the world in which we live, and an increasing concern for our own health. The delicate balance of the natural world and strive to maintain this harmony by using environmentally-friendly products that are nurturing to us and as harmless as possible to the earth. Natural oils are vegetable oils consist of aethereal salts of glycerin with a large number of organic acids such as stearic acid, oleic acid, and palmitic acid forming stearin, olein and palmitin, respectively. Natural oils are excellent emollients leaving the skin soft and smooth. While penetrating the skin many oils have also effective nourishing and revitalizing effects. Natural oils are used in a wide variety of cosmetic products including personal care as well as makeup products. The contents of bioactive lipophilic compounds promote elasticity, healing, moisturizes and helps the skin maintain a proper moisture balance and protects the skin from free radical damage. Natural oils of olive, jojoba seeds, castor seed, cedar balsam, grape seeds and sea buckthorn fruits were chosen as potent active constituents for lipstick production. Antioxidant activity was evaluated by and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical inactivation method spectrophotometrically. There is the sequence of results: 68% sea buckthorn oil, 49.03% grape seed oil, 31.13% castor oil, 15.74% olive oil, 9.03% jojoba oil and 3.92 castor balsam oil.

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P-37 ANTIFUNGAL ACTIVITY OF PHENOLIC COMPOUNDS FROM FAGOPYRUM ESCULENTUM AGAINST FUSARIUM CULMORUM Ilona Kerienė1, Audronė Mankevičienė1, Bronislava Butkutė1, Audrius Maruška2 1

Institute of Agriculture, Lithuanian Research Centre for Agriculture and Forestry Instituto alėja 1 Akademija LT-58344 Kėdainiai distr. Lithuania E-mail: [email protected] 2 Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos 8, LT-44404 Kaunas, distr. Lithuania

Abstract The antifungal activity in vitro of phenolic extracts of buckwheat (Fagopyrum Esculentum Moench) was tested on important species of Fusarium culmorum (W.G. Smith) Sacc., which was isolated at the Department of Plant Pathology and Protection. The extracts from buckwheat grain, groats and hulls with the phenolics concentrations of 3.15 mg ml-1 ; 1.28 mg ml-1 ; 1.48 mg ml-1, respectively; as well as pure rutin and quercetin solutions at 7.52; 3.76; 1.2 mg ml-1 concentrations each, were used in the test. A small segment (0.5 cm) of the fungal growth was transferred to PDA medium, incubated at 22±2°C and the extension diameter (cm) of hyphae from centre to the sides of dishes was measured periodically for 8 days. Ethanol (96 %), without extract solutions, was used as control. The results were subjected to ANOVA using Fisher’s LSD test (p< 0.05). In all samples, significant differences in antifungal activity were detected after 96 hours incubation. Quercetin showed the strongest inhibitory effect (40%) on the mycelia growth, which lasted from the 96th hour of incubation until the end of the test (192 hours). Expansion area of fungal mycelium was 5.54±0.46 cm, compared with the control (8.60±0.05 cm). A significant inhibitory effect on F. culmorum was observed with extracts of groats and hulls. After 192 hours incubation, the area of expansion accounted for 7.39±0.49 cm and 7.19±0.37 cm, respectively. Antifungal activity of the phenols extracted from buckwheat grain was insignificant. Significant antifungal activity of rutin lasted for 144 hours. However, the differences between concentrations of quercetin and rutin in the solution did not have any influence on mycelium growth inhibition.Our research indicated no direct relationship between the higher concentration of phenols and antifungal activity, because a pure phenol compound quercetin exhibited a greater antifungal activity. It is likely that biologically active compounds or their group can effectively act in the extracts of groats and hulls. A buckwheat variety might be a significant factor, too. To validate this assumption, the research needs to be continued. Acknowledgements The abstract presents research findings obtained through the long-term research programme “Harmful organisms in agro and forest ecosystems” implemented by Lithuanian Research Centre for Agriculture and Forestry.

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P-38 DETERMINATION OF PHENOLIC AND ANTIOXIDANT PROFILES OF LINDEN (TILIA CORDATA MILL.) INFLORESCENCES Raimondas Raudonis1,2*, Lina Raudonė1, Valdimaras Janulis1, Pranas Viškelis2 1

Department of Pharmacognosy, Lithuanian University of Health Sciences, Mickeviciaus str. 9, LT-44307, Kaunas, Lithuania 2 Institute of Horticulture, Lithuanian Research Center for Agriculture and Forestry, Kauno str. 30, LT54333, Babtai, Kaunas distr., Lithuania;*E-mail: [email protected]

Linden (Tilia L.) inflorescences have a prominent importance in phytotherapy [1]. Tilia L. extracts has a long history of usage for the treatment of bronchitis, influenza, inflammation and anxiety. Inflorescences of linden possess diuretic, diaphoretic, antipyretic, anti-inflammatory as well as tranquilizing activities [1,2]. Principle secondary metabolites of Tilia L. responsible for these therapeutic activities are flavonoids and phenolic acids. They are regarded as natural antioxidants that play an important role in protecting cells from oxidative stress [3]. The determination of antioxidant profile and investigation of contribution of different phenolic compounds to antioxidant activity is important for the evaluation of Tilia L. extracts. Therefore, the aim of this work was to determine the phenolic and antioxidant profiles of Tilia cordata Mill., inflorescences and to identify markers of antioxidant activity. Ethanolic (70% v/v) extracts of Tilia cordata Mill., inflorescences were analyzed using FRAP post-column assay. Antioxidant active compounds were quantified according to Trolox equivalent (TE) for each assay. Results demonstrate that Tilia cordata Mill., extract possessed strong antioxidant activity, TE=10.89 mg/g (dry weight). Eleven phenolic compounds (gallic acid, protocatechuic acid, coumaric acid, rutin, hyperoside, avicularin, quercitrin, astragalin, isoquercitrin, tiliroside, quercetin) were identified in the chromatographic profile of Tilia cordata Mill., inflorescence extracts. In the antioxidant profile 30 compounds of different activity were determined. Assessed TE values confirm that protocatechuic acid (TE=0.87 mg/g (dry weight)) and gallic acid (TE=0.32 mg/g (dry weight)) had the greatest antioxidant activity from the identified compounds. The contribution of protocatechuic and gallic acid to total antioxidant activity was 8.13 % and 2.94 %, respectively. The dominant compound of chromatographic profile tiliroside had no significant antioxidant activity. Summarized experimental results revealed that protocatechuic acid can be used as marker for the evaluation antioxidant activity of inflorescences of Tilia cordata Mill. References: 1. Toker G, Aslan M, Yesilada E, Memisoglu M, Ito S (2001) J Pharm Biomed anal 26:111-121 2. Marrassini C, Anesini C, Ferraro G (2011) Phytother Res 25:1466-1471 3. Aguirre-Hernandez A, Gonzales-Trujano ME, Martinez AL, Moreno J, Kite G, Terrazas T, Soto-Hernandez M (2010) J Ethopharmacol 127:91-97

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P-39 BOTANICAL COMPONENTS AND RAW MATERIALS OF ANIMAL ORIGIN IN THE PRESCRIPRION BOOK FROM 1830 Zenona Šimaitienė, Vilma Gudienė, Tauras Mekas The Museum of the History of Lithuanian Medicine and Pharmacy

The Museum of the History of Lithuanian Medicine and Pharmacy has a prescription book by a pharmacist T. Göldner from Telšiai (the tsarist Russia’s Lithuanian Vilnius Governorate), dated 1830. Each medication that was produced at the pharmacy had to be registered in this book. The entries included the composition of the drug, its form, usage, price, the physician’s name, and (sometimes) the method of production (1). Botanical components were most commonly used – they comprised 71% of all the components. In total, the pharmacist used over 100 plants. Both local and imported botanical raw material was used for drug manufacturing. Poppy syrup, poppy flowers, and opium are mentioned in the composition of every fifteenth drug produced at Telšiai pharmacy. Henbane (Hyoscyamus) extract was used for the production of internal medications, and oil – for external drugs. At the beginning of the 19th century, medicines containing substances of animal origin were still popular. In the prescription book of Telšiai pharmacy, such components comprised 6% of all ingredients. Most frequently, adhesive plaster Emplastrum Vesicatorii was prescribed, containing powder of Spanish flies (Cantharides), extract and tincture of beaver glands (castor sacs) (Castoreum), bee products, and – less frequently – substances obtained from bulls, deer, crayfish, or ants. Patient treatment was based on the principles of the humors theory. To restore the balance of fluids (humors), physicians prescribed diuretics (Diuretica), emetics (Emetica), laxatives (Laxantas), diaphoretics (Diaphoretica), expectorants (Expectorantia), etc. (2). References: 1. Archives of Lithuanian Museum of the History of Medicine and Pharmacy, Kaunas (1830a) Shnurovaja kniga Provizora Fedora Geldnera v gorod Telshiach sostojashchiuju, 1830 goda. (Pharmacist Theodor Göldner’s Prescription Book for 1830 in Telšiai City 2. Shprengel K (1820) Lekarstvenik ili farmacologia Kurta Shprengelia (Kurt Sprengel’s Drug Registry, or Pharmacology), Moscow.

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P-40 SUSCEPTIBILITY OF E. COLI TOWARDS CHITOSAN MODIFIED BY SPECIFIC INORGANIC AND ORGANIC BIOCIDES Iglė Vepštaitė1,2, Juliana Lukša1, Ona Gylienė3, Rima Binkienė3, Elena Servienė1,2 1

Nature Research Centre, 2Vilnius Gediminas Technical University, 3Center for Physical Sciences and Technology

Chitosan is a cationic polysaccharide obtained by alkaline deacetylation of chitin, a naturally occurring polymer. This biopolymer has a profound potential for diverse applications due to its biodegradability, biocompatibility, antimicrobial activity, non-toxicity and versatile chemical and physical properties [1]. It has been found effective in inhibiting growth of variety Gram-positive and Gram-negative bacteria, thus showing great promise for application in food preservation [2]. The present study examines the effect of chitosan samples covering wide range of molecular weight, degree of deacetylation, organic and inorganic modifications, on ability to affect the growth of E. coli. Sensitivity and viability of model prokaryotic bacteria towards dissolved or suspended chitosans were tested. It was observed that non-modified samples demonstrate weak antibacterial properties. Introduction of inorganic or organic modifications increase antibacterial properties, and profound effect was observed in case of double modified chitosans. References 1. Chitosan derivatives obtained by chemical modifications for biomedical and environmental applications: Alves NM, Mano JF (2008) Biol Macromolec 43: 401-414. 2. Progress in antimicrobial activities of chitin, chitosan and its oligonucleotides: a systematic study needs for food application: Dutta J, Tripathi S, Dutta PK (2012) Food Science and Technol Int, DOI: 10.1177/1082013211399195

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P-41 BORRELIA SPP. GENOTYPING IN SMALL RODENTS AND TICKS Baliutytė Vaida, Radzijevskaja Jana Departament of Biology, Vytautas Magnus university, Kaunas, Lithuania

Borrelia spp. is a bacterial species of the spirochete class of the genus Borrelia. The genus Borrelia usually is divided into two groups: causing recurrent fever and another group that is associated with Lyme disease. The latter group consists of B. burgdoferi sensu lato complex (Chinmoy and et al., 2011). Different Borrelia spp. species can be associated with different reservoir hosts: B. afzelii with Yellow-necked Field mouse (Apodemus flavicolis) B. burgdorferi with red squirrels (Sciurus vulgaris) and B. garinii with some bird species. Clinically and ecologically important, what are Borrelia spieces prevalent in a particular area. It is also important to accurately determine the Borrelia species and strains of different organisms and tissu (Radzijevskaja et al., 2011). 481 I. ricinus and I. persulcatus ticks were analyzed from seven different locations in Estonia and 72 rodent ears, and urinary bladder in six different locations in Lithuania. Our studies have used genome plasmid ospA gene, fla gene and the ribosomal intergenic spacer (IGS) region molecular markers. Studies have shown that more sensitive and appropriate pathogen detection was nested PCR, which amplified IGS molecular marker. Using this marker, it was found that a total of 24 (33%) rodents were infected Borrelia spp. pathogens, while using ospA marker, only 13 (18%). In rodents were found B. afzelii and B. miyamotoi species. Also rodent samples were tested for B. valaisiana pathogen infection, but the bacteria were not detected in the tested samples. Using ospA marker only 6 rodents Borrelia spp. pathogens have been identified in ears, and urinary bladder samples, three rodents were positive only by ear samples, four rodents - just under the bladder samples. According to IGS molecular markers, six rodents were positive by ear, and urinary bladder samples, 9 rodents under the ear and under the bladder samples. Most infected ticks was found in the three regions of Estonia, where is the biggest infection of human. Out of 481 ticks, 76 were infected with Borrelia spp. pathogen. In all infected tic were found two pathogen species: B. afzelii and B. garinii. Keywords: Borrelia spp., small rodents, ticks. 1. Chinmoy B., Robert A. Schwartz I. 2011. Lyme disease, Part I. Advances and perspectives. MPH Newark, New Jersey, J Amakad dermatol;64(4): p. 619-18. 2. Radzijevskaja J., Paulauskas A., Rosef O. Molecular detection and characterization of Borrelia burgdorferi sensu lato in small rodents. Veterinarija ir zootechnika, 2011; 55(77): p. 40-47.

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P-42 INTRODUCTION AND PHYTOCHEMICAL ANALYSIS OF VIOLA TRICOLOR L. GROWN AT KAUNAS BOTANICAL GARDEN OF VYTAUTAS MAGNUS UNIVERSITY Béquet Karla 1,2,3, Audrius Maruška3, Ona Ragažinskienė2 1

Angers’s University, Institute of Technology (IUT),4 bld Lavoisier - BP 42OI8-49016A NGERSC EDEX; Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Ž.E. Žilibero str. 6, LT- 46324 Kaunas, Lithuania 3 Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos str. 8, LT-44404, Kaunas, Lithuania 2

The research work presents details of introduction of Wild pansy (Viola tricolor L.) since 1986 in collection of medicinal plants of Sector of Medicinal Plants at Kaunas Botanical Garden of Vytautas Magnus University (VMU) and its phytochemical analysis results in Instrumental analysis laboratory at Department of Biochemistry and Biotechnologies of VMU. The raw material of this medicinal plant was collected and dried in 2012. Viola tricolor L. is an annual herbaceous plant, belonging to Violaceae Batsch family. The plant is rich in biologically active compounds, such as polyphenols (flavonoide). The aim of the studies was to determine the polyphenols (flavonoide), content and the total scavenging activities in the dried flowering aerial parts Viola tricolor L. – Violae herba cum flore. Liquid extraction with aqueous methanol was carried out for spectrophotometric analysis of total amount of phenolic compounds, total amount of flavonoids and DPPH radical scavenging activity. Phenolic compounds were identified and quantitated using high performance liquid chromatography. For assessment of radical scavenging of individual phenolic compounds the online post-column DPPH reaction detection was carried out. The obtained results demonstrate the variety and complexity of the biologically active compounds present in Viola tricolor L grown in the Middle Lithuania climatic conditions. The results suggest that majority of compounds are in the blossom and the major is rutin which have a primordial influence in the anti-oxidant activities. Keywords: Viola tricolor L., introduction, phytochemical analysis, Lithuania. Direction Européenne de la Qualité du Médicament & des Soins de Santé. European Pharmacopoeia 6.0. Council of Europe (EDQM). Wild Pansy (flowering aerial parts), 2008 p. 2700-2701 Guttman.A, Guenther.K, Kerry.A , Ringer. T, Vukics .V. Analysis of Heartsease (Viola tricolor L.) flavonoid, glycosides by micro-liquid chromatography coupled to multistage mass spectometry. Journal of Chromatographic A, 2005, N°1206, p. 11-20. Athayde.M, Boligon.A, Beck.R, Gonçalves.A, Piana.M . Antioxydant capacity, total phenolic contents and HPLC determination of rutin in Viola tricolor L flowers. Free radical and antioxydants, Vol 2, oct-dec 2012, p.32-37. Rimkiene S. Ragazinskiene O. Savickiene N. The cumulation of Wild pansy (Viola tricolor L.) accessions: the possibility of species preservation and usage in medicine. Medicina (Kaunas) 39(4):411-6, 2003.

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P-43 COMPARATIVE DEVELOPMENT AND EVALUATION OF BAIKAL SCULLCAPS (SCUTELLARIA BAICALENSIS GEORGI) TOPICAL GELS, GEL – CREAMS AND CREAMS FORMULATIONS Vytis Čižinauskas, Vitalis Briedis, Kristina Ramanauskienė Lithuanian University of Health Sciences, Faculty of Pharmacy, Department of Clinical Pharmacy

Abstract Baikal skullcap (Scutellaria baicalensis Georgi) is one of the main medicinal plants of traditional Chinese medicine and is gaining more interest in researches, because of its flavonoids activities. And for the present there are none scientific examination of its topical preparations. This work aims on investigation of Baikal skullcap extract incorporation into different semi-solid dosage and biopharmaceutical evaluation. Three types of topical preparations with 1% of Baikal skullcaps extract were prepared: carbomeric hydrogels using 0.5% - 1% Carbopol 980 NF; gel – creams using different amounts of Poloxamer 407 and Kollisolv® systems; creams using comercial Pionier® PLW and Pionier® PLW PAO E bases. Viscosity of these forms was tested. In order to properly perform biopharmaceutical evaluation, modified Franz cells with cellulose membrane and LC – DAD method for identification and quantification of main flavonoids was used. The results showed that gels viscosity influences the release of active compounds from the system and the biggest amount of flavonoids was released from a system, which was made with 0.5% of Carbopol 980 NF. Comparing all systems gel – cream composition with 15 % of Poloxamer 407 releases the highest amount of measured flavonoids during 6 hours 10.20 ± 0.49% respectively. While both creams showed more than 20 times lower release of flavonoids comparing to previously mentioned gel-cream composition. Considering drug release and rheological properties gels and gel – creams are promising form for biopharmaceutical evaluation on skin. Keywords: Baikal skullcap, Scutellaria baikalensis Georgi, biopharmaceutical evaluation, in vitro evaluation, carbomer gels, gel – creams, creams, semisolids, topical dosage forms References 4. Universités de Médecine Traditionnelle Chinoise de Nanjing et Shanghai (2008). La pharmacopée chinoise. Les herbes médicinales usuelles. 中药学, Editions You Feng. (ISBN 978-2-84279-361-7). 101 – 103 5. Evans W. C. (2009) Trease and Evans Pharmacognosy 16th Edition. Edinburgh London New York Philadelphia St Louis Sydney Toronto. 6. Leach F. S. (2011) Anti-microbial properties of Scutellaria baicalensis and Coptis chinensis, two traditional Chinese medicines. Bioscience horizons. Vol. 4, No 2. 119 – 127

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P-44 DIFFERENT GROWTH RESPONSES OF PISUM SATIVUM AND MELILOTUS ALBA TO 700 AND 1500 ppm OF CARBON DIOXIDE Austra Dikšaitytė, Irena Januškaitienė Vytautas Magnus University

The aim of this work was to determine the growth responses of C3 legume crop pea (Pisum sativum L.) and weed white melilot (Melilotus alba Medik.) at the early stage (until 27 days after sowing) under current (350 ppm) and elevated (700 and 1500 ppm) CO2, in order to assess, how will respond a crop and a weed of the same family in monoculture conditions to future elevated CO2 concentrations. Experiments were conducted in the closed environment-controlled chambers. Duration of the experiment: 10 days. Measurements of physiological plant indices – photosynthetic rate (A), stomatal conductance (gs), transpiration rate (E), water use efficiency (WUE) and the ratio of intercellular to atmospheric CO2 concentrations (ci/ca) were carried out on the last day of treatment. Biochemical analysis (foliar and roots carbohydrates) and growth response (shoots and roots dry weight) of plants were carried out at the end of experiment. 700 and 1500 ppm elevatedCO2 increased photosynthetic rate by 46 and 66 % (p < 0.05) of pea plants and reduced gs by 53 and 39 % and E by 38 and 28 % (p < 0.05), which resulted in 1.4 and 1.3 times greater WUE, respectively, compare to pea plants gown in current-CO2. Despite significant decrease in gs the ci/ca ratio increased by 14 and 28 % (p < 0.05), respectively under 700 and 1500 ppm CO2, so this indicate, that plants grown in 700 and 1500 ppm CO2 environments had a higher intercellular CO2 concentration and the decrease of gs did not limited photosynthesis. Differently gs of white melilot plants grown at 700 and 1500 ppm CO2 increased by 39 and 54 % (p < 0.05) respectively, and E and ci/ca ratio also increased by 21 and 26 %, and 34 and 59 % (p < 0.05), respectively, compare to plants grown in current CO2. Whereas A was up-regulated significantly by 15 % only at 1500 ppm CO2, and this resulted in 16 and 9 % (p < 0.05) reduce of WUE, respectively, compare to current CO2. The total non-structural carbohydrate (TNC) decreased significantly by 43 % in leaves and 34 % in roots under 700 ppm CO2 and increased by 44 and 276 % (p < 0.05) in leaves and roots, respectively, in pea plants grown in 1500 ppm elevated-CO2, compared to current-CO2 pea plants. While significant change in TNC of elevated-CO2 white melilot plants were detected only at 1500 ppm CO2, when TNC level in roots were 26 % lower than reference treatment. 700 and 1500 ppm CO2 had no significant effect on shoots and roots dry weight of weed. While 700 elevated-CO2 increased shoots and roots dry weight of pea plants by 10 and 3 % (p < 0.05), respectively, and 1500 CO2 increased significantly shoots dry weight by 8 %, compare to pea plants grown in current-CO2. Thus under elevated 700 and 1500 ppm CO2 the ability of pea to assimilate additional carbon is higher, compared to white melilot – a weed of the same family.

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P-45 BOUND STATES OF FEW NUCLEON SYSTEMS Saulius Mickevičius Vytautas Magnus University, K. Donelaičio 58, LT-44248, Kaunas, Lithuania [email protected]

A method for lower bounds calculation for the lighest atomic nuclei is introduced. The effiency of this method, which is based on factorization of the antisymmetrizer operator, is compared to the direct one.The precision of the method is teste in three and four-body calculations. Nucleons are invariant with respect to translations in space, therefore the wave functions of the selfbound systems must be translationally invariant. The best-known methods for the description of quantum systems, such as the Shell Model or the Hartree-Fock Self-Consistent Field method, produce wave functions dependent on a set of one-particle variables, thus also on the center of the mass radius-vector of the system. This shortage of mentioned methods is well known, however, the wave functions dependent on one-particle variables are very attractive because they allow a simple procedure of antisymmetrization. In our calculations [1] we use the normalized Jacobi coordinate system, known as Jacobi tree, to ensure translational invariance of many-body wave function. References 1. Germanas D., Kalinauskas R. K., Kamuntavičius G. P., Žemaičiūnienė R. Large-Basis HarmonicOscillator Shell Model Application to Alpha Particle. Lithuanian Journal of Physics 44, 2004, 243-248 p.

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P-46 EFECTIVE CALCULATION OF HARMONIC OSCILLATOR TRANSFORMATION BRACKETS S. Mickevičius Vytautas Magnus University, K. Donelaičio 58, LT-44248, Kaunas, Lithuania [email protected]

The wave function of a self-bound system in the absence of external fields must be invariant in respect of spatial translations as well as antisymmetric in respect of all permutations of the fermions. One of the possibilities to find a solution to the problem of translational invariance of any wave function is based on direct construction of the many-fermion wave function, which is independent of the center of mass coordinate [1]. In this case, the harmonic-oscillator (HO) basis set in terms of intrinsic (Jacobi) coordinates is necessary. Only the set of HO functions can be chosen in such a manner that the transformation from one set of Jacobi coordinates to other results in a corresponding expansion with a finite number of terms. In this approach, the essential feature is the Talmi-Moshinsky transformation [2] and corresponding HO brackets. Since HO brackets are constantly employed in various model calculations of the nuclear and hadron structure, it is desirable to have a simple and efficient method to calculate them [3]. References D. Germanas, S. Mickevičius, R.K. Kalinauskas, Calculation of four-particle harmonic-oscillator transformation brackets, Computer Physics Communications 181, 420--425 (2010). S. Mickevičius, E. Brazauskas, D. Germanas, R.K. Kalinauskas, The four-particle harmonicoscillator brackets: Compact expressions and updated Fortran program, Computer Physics Communications 182, 1377-1381 (2011). S. Mickevičius, D. Germanas, R.K. Kalinauskas, Four particle harmonic-oscillator transformation brackets and their simplifications for special values of the mass-ratio parameters, Central European Journal of Physics, Vol. 181, No. 2 (2012).

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P-47 EVALUATION OF SOLVENT INFLUENCE OF THE QUALITY OF ETHANOLIC EXTRACTS OF SAMBUCUS NIGRA L FRUITS Saulė Velžienė, Lauryna Pudžiuvelytė, Giedrė Kasparavičienė, Zenona Kalvėnienė Department of Drugs Technology and Social Pharmacy, Faculty of Pharmacy, Medical Academy, Lithuanian University of Health Science A. Mickevičiaus 9, LT – 44307 Kaunas, Lithuania

Black elderberry (Sambucus nigra L.), is employed as an alternative to conventional medicines and mainly in the form of an extract for treating the common cold, influenza and Herpes virus infections. Elderberry is often recommended for use as a complementary therapy together with the classic antioxidant nutrients, vitamin C and zinc, to support the natural process of recuperation. The European black elderberries are rich dietary sources of plant pigments and phenolic compounds. Ethanolic extracts (30%, 50%, and 70%) of elderberry were analyzed using spectrophotometric methods for total amount of anthocyanins (expressed by Cyd-3-glu equivalents) and polyphenols (expressed by gallic acid equivalents) and evaluation of antioxidant activity by and 2,2-diphenyl-2picrylhydrazyl (DPPH) radical inactivation. Increasing concentration of ethanol from 30% - 50% - 70% extracted increasing amount of total phenolics 3.18 – 4.06 – 5.29 mg/ml GAE, 34.58% - 35.69% - 52.60% inactivated DPPH also, but amount of anthocyanins varied differently 89.90 – 64.49 – 269.48 mg/L. 70% concentration of ethanol can be considered as the most potent solvent concentration for the extraction of Sambucus nigra L fruits.

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P-48 THE APPLICATION OF ADMS-URBAN MODEL TO ESTIMATE NITROGEN DIOXIDE CONCENTRATIONS IN COLD AND WARM SEASONS IN KAUNAS CITY Audrius Dėdelė, Auksė Miškinytė Vytautas Magnus University, Department of Environmental Science

Abstract In urban areas road traffic is one of the main sources of pollution affecting air quality. Nitrogen dioxide (NO2) is one of the most common traffic-related pollutants. ADMS-Urban can be used to examine emissions from many various sources (road transport, industrial, domestic). The paper examines the performance of ADMS-Urban dispersion model to predict nitrogen dioxide concentrations from emission sources. Modelled concentrations for cold and warm seasons have been compared with measured nitrogen dioxide concentrations at 41 sites in Kaunas city. The purpose of this study was to assess the dispersion of nitrogen dioxide pollution in cold and warm seasons comparing modelled with ADMS-Urban model and measured concentrations in Kaunas city. Modelled average nitrogen dioxide concentration in cold season was 21.79 μg/m3, while measured with passive samplers – 19.82 μg/m3. The air pollution was significantly higher during the cold season. Modelled and measured NO2 concentrations in warm season were 12.28 and 11.62 μg/m3, respectively. Results showed that ADMS-Urban dispersion model tends to overpredict nitrogen dioxide concentrations and this is most evident when smaller values are modelled, while modelled maximum concentrations are underestimated comparing with measured NO2 values.

Keywords: nitrogen dioxide, modelling, ADMS-Urban model, cold and warm seasons

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P-49 EXTRACTION PROCEDURE OPTIMIZATION OF ROSELLE (HIBISCUS SABDARIFFA L.) CALYCES Gintarė Sokaitė1, Zenona Kalvėnienė1, Giedrė Kasparavičienė1, Audrius Maruška2 1

Lithuanian University of Health Sciences, Medical Academy, Faculty of Pharmacy, Department of Pharmaceutical Technology and Social Pharmacy. A. Mickevičiaus 9, LT44307 Kaunas 2 Vytautas Magnus University, Faculty of Natural Sciences, Department of Biochemistry and Biotechnologies.Vileikos 8, LT44404 Kaunas

Roselle (Hibiscus Sabdariffa L.) calyces is a medicinal herbal material described in European Pharmacopoeia also known in Eastern medicine for its antihypertensive, antiinflammatory, diuretic, atherosclerosis reducing and cancer preventive effects. Pharmacological effects are addressed to polyphenolic compounds present in roselle. Aqueous extract color is intensively red; intensity represents the extent of extracted anthocyanins, mainly, cyanidin and delphinidin. The aim of this work was optimization of aqueous extraction procedure of roselle calyces. Classical and modern extraction methods were applied in order to prepare roselle extracts. Magnetic stirrer, ultrasound, microwave assisted and pressurized hot water extraction modes were used. The quality of prepared extracts was evaluated spectrophotometrically by determining coloring power, total phenolic compounds and total flavonoid content and evaluating radical scavenging activity. Total phenolic compound content was determined by Folin-Ciocalteu method. Total flavonoid content was determined by colorimetric method in the presence of aluminum chloride. Radical scavenging activity was evaluated using free DPPH• radical. Coloring power was determined by method, described in European Pharmacopoeia. Highest recovery of pigments was obtained by pressurized hot water extraction. This method also provided highest recovery of phenolic compounds (3.12 mg GAE/ml ± 1.8%) and flavonoids (0.877 mg RE/ml ± 3.73%). Highest radical scavenging activity was observed in extracts prepared with the aid of magnetic stirrer for 30 min. at 80°C temperature (0.198 mg AAE/ml ± 7.41%).

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P-50 ANTIOXIDANT PROPERTIES AND ELEMENT CONCENTRATIONS OF THE FLOWERS OF IMPATIENS GLANDULIFERA Eglė Kazlauskaitė, Irma Šliumpaitė, Lina Zybartaitė, Algimantas Paulauskas, Eugenija Kupčinskienė Faculty of Natural Sciences, Vytautas Magnus University, Vileikos 8, LT- 44404 Kaunas, Lithuania,

Since long time flower morphology of Impatiens species was subject of wide investigations. Impatiens glandulifera is an alien invasive plant with huge and bright flowers. Flowering time of this species usually starts in July and lasts till the frosts. For I. glandulifera five types of flowers are documented according to colour which ranges from gentle pink to dark purple. Till now insufficient attention has been paid to the physiology and biochemistry of this part of the plant which is attractive for both insects and human beings. Our study aimed at evaluation of variety of I. glandulifera populations according to the concentrations of antioxidants and elements in the flowers. Plant material was collected from thirty locations in Lithuania. After transportation to the laboratory different parts of the plants were separated, Material was dried till the permanent weight and grinded with the mill to obtain the homogenic powder. Flower extracts were prepared using 3 types of solvents (water, ethanol, butanol) using cold extraction. For the measurements of the antioxidant activity of the extracts, 1,1-diphenyl-2picrylhydraryl radical (DPPH) and 2,2‟-azinobis(3-ethylbenzothiaziline-6-sulfonate) ABTS were used, estimating the free radical scavenging activity. Also total phenolic content (TPC) was evaluated using Folin-Ciocalteu method. To evaluate concentrations of the nutrients and trace elements in the flowers, flame atomic-absorption spectrophotometry and graphite furnace atomic absorption spectrometry were applied. For element analyses flower powder was digested in by the mixter of 65 % HNO3 and 30 % H2O2. Prepared extracts were atomized in the flame of acetylene and weather mixture in ~2300 °C. Quiet high concentrations of phenolic compounds were documented, at the same time free radical scavening activity was not high. Among investigated elements the highest concentrations of potassium and magnesium were characteristic for the flowers. Among trace elements the lowest concentration was for cadmium. The obtained results showed that Impatiens glandulifera flowers had lower antioxidant activity and smaller amounts of mineral elements when compared to the other organs.

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P-51 GLUTAMIC ACID LEVELS IN MEAT PRODUCTS K. Barčauskaitė1, G. Garmienė1, I. Miliauskienė1, I. Jasutienė1, A. Šalaševičienė1 1 Kaunas University of Technology, Food Institute, Taikos pr. 92, LT-3031 Kaunas

Abstact Producers supply to the market meat products, produced without the flavor enhancer, but the free glutamic acid may occur naturally in protein containing products. Free glutamic L (+) acid content in meat products depends on the protein content and technological parameters, maturation and storage time. The aim of work is to analyze of free glutamic L (+) acid levels in meat products, according labeling produced without monosodium glutamate E621, present in Lithuanian market. After testing 60 samples of cooked sausages, the highest content of free glutamic L (+) acid was found 0.95 and 0.75 g/kg, the average level was found to be (0.18 ± 0.15) g/kg. The level of free glutamic acid in hot smoked sausage samples vary from 0.08 to 0.50 g/kg, the average level was found to be (0.19 ± 0.11) g/kg. After testing smoked sausage samples, the average glutamic L (+) acid content was (0.46 ± 0.30) g/kg, but this value varied from 0.07 to 0.80 g/kg. The highest content of free glutamic L (+) acid was found in dried sausage samples - the average value was found to be (0.82 ± 0.43) g/kg, the lowest content was 0.25 g/kg, while the highest - 1.27 g/kg. The highest content of protein are in dried sausages, in these samples was found and the highest content of glutamic L (+) acid, but between individual samples this parameter ranged up to 5 times. Keywords: glutamic acid, sausage, meat products

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P-52 ANISE HYSSOP INTRODUCTION AND THIS MEDICINAL PLANT RAW MATERIAL IMPORTANCE FOR HUMAN HEALTH Jovita Bagočiūtė1, Mantas Stankevičius1, Kristina Bimbiraitė-Survilienė1, Olga Kornyšova1, Vilma Kaškonienė1, Ona Ragažinskienė2, Audrius Maruška1 1

Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos str. 8, LT44404, Kaunas, Lithuania 2 Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Ž.E. Žilibero str. 6, LT- 46324 Kaunas, Lithuania

Pharmacological effect of medical plant material on humans depends on its biologically active substances. Therefore, it is also important to investigate introduced plants that do not grow naturally in Lithuanian territory and which can be used as medical plant material because of its positive effects on human. This research work was to analyse the quality of medicinal plant material of introduced anise hyssop. Research object - anise hyssop (Agastache Foeniculum (Pursh) Kuntze) is a perennial herbaceous plant belonging to Lamiaceae Lindl. family. It is a medicinal, herby, melliferous, and decorative plant that originates from North America, but still not widespread in Lithuania. It is being introduced by Kaunas Botanical Garden in Vytautas Magnus University since 1972. Anise hyssop is a natural of biostimulator which can act in antibacterial way, it lowers blood pressure, and alleviates cough. Preparations from Agastache foeniculum are used in folk medicine to improve metabolism, the infusions are used to strengthen the hair, in perfume industry and in gastronomy, as a spice. The subject of method is preparation of methanol extracts with 75% MeOH, high performance liquid chromatography with reaction detector (HPLC), a solid phase microextraction and gas chromatography with mass spectrometry (GC-MS). The main phenolic compounds were identified in the plant after the HPLC with the detector: chlorogenic acid, rutin and quercetin. Main essential oils were identified in the plant after gas chromatography with mass spectrometry: D-limonene, estragol, caryophyllene and germacren D. Phenolic compounds that are beneficial to the human body because of antioxidative, antiinflammatory and antiviral properties were identified in anise hyssop. All the essential oils that were found are used in food production, as spices and in fragrance industry.

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P-53 ANALYSIS OF HUMULUS LUPULUS L. ESSENTIAL OILS AND BITTER ACIDS Paulius Orentas1, Žydrūnas Stanius1, Paulius Kaškonas2, Tomas Drevinskas1, Mantas Stankevičius1, Kęstutis Obelevičius3, Ona Ragažinskienė3, Audrius Maruška1 1Vytautas Magnus University, Department of Biochemistry and Biotechnologies, Vileikos st. 8, LT44404 Kaunas, Lithuania 2Kaunas University of Technology, Metrology Institute, Studentų str. 50, LT-51368, Kaunas, Lithuania 3Kaunas Botanical Garden of Vytautas Magnus University, Ž.E.Žilibero 6, LT-46324 Kaunas, Lithuania

The object of this research is Humulus lupulus L. one of the many medicinal plants, which has wide application in beer production industry. This is aromatic, multi-climber common hop (Humulus lupulus L.), which is growing throughout the territory of Lithuania. Traditional medicine knew hops cones in VIII century and used for insomnia, as nerve sedative, improving digestion, stimulating the release of urine, pain and seizures depressing measure of cough, fever, diarrhea, inflammation, and rheumatism. It is known that hop cones have antibacterial, fungicidal properties and estrogenic activity. Hop cones accumulate essential oil (1.0-3.0%) containing β-pinene, β-myrcene, caryophyllene, α-caryophyllene and other volatile compounds. Important biologically active compounds group is bitter accids (5.0-20.0%): alpha-bitter acid (humulone), beta-bitter acid (lupulone), enzymes, flavonoids (rutin, isoquercitrine etc.), anthocyanins (0.3%), leukoantocianidine, ascorbic acid, and vitamin B1. The aimof this study was to determine and compare the 2010, 2011 and 2012 harvests and different varieties of hops (Humulus lupulus L.) essential oils composition and bitter acid compositions using gas chromatography-mass spectrometry and capillary electrophoresis methods. For the analysis raw material of common hop (Humulus lupulus L.) cones was collected at Kaunas Botanical Garden of Vytautas Magnus University hops collection. The raw material of collected cones was dried in a well ventilated room, avoiding direct sunlight. The highest amount of essential oils was determined in hop “Marynka” were identified: αpinene, β-mircenae; humulene; caryophyllene. This variety of hops was rich with alpha-bitter acid (co-humulone, and ad-humulone) and beta-bitter acid (lupulone). Keywords: bitter acids, essential oils, hops, CE, GC-MS

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P-54 VARIATION OF TOTAL AMOUNT OF PHENOLIC AND VOLATILE COMPOUNDS OF SATUREJA HORTENSIS L. AND SATUREJA MONTANA L. EXTRACTS DURING DIFFERENT VEGETATION PERIODS Andrius Rimovas1, Ieva Akuneca1, Mantas Stankevičius1, Olga Kornyšova1, Ona Ragažinskienė2, Audrius Maruška1 1

Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos str. 8, LT44404, Kaunas, Lithuania 2 Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Ž.E. Žilibero str. 6, LT- 46324 Kaunas, Lithuania

Savory – belongs to Lamiaceae family of plants. It is an annual herb used as a spice. They are ca. 30 species, however in Lithuania is are growing only two: Winter savory (Satureja montana L.); savory (Satureja hortensis L.). Savory has a strong, pleasant thyme-like aroma, tangy taste. Ethanolic extract and essential oil prepared of raw material of savory upper-ground part is used in the treatment of stomach, intestine, kidney, liver diseases, acts as an antispasmodic, bactericidal, appetite increasing preparation. The aim of this study was to carry out phytochemical analysis of savory at different growth stages, to determine radical scavenging activity, total amount of phenolic compounds and the composition of essential oils. Results: Using gas chromatography-mass spectrometry analysis the Satureja hortensis L. and Satureja montana L. essential oil components were identified. The highest concentrations of carvone, gamma-terpinene, o-cymene, caryophyllene, β-bisabolene were determined. It was found that in the Satureja hortensis L. decrease of gamma-terpinene and increase of antioxin concentration during vegetation is observed. In Satureja montana L. the decrese of gammaterpinene during the vegetation was determined. Spectrophotometric analysis revealed, that of total amount of phenolic compounds in Satureja montana L. ranged from 45.4 mg/g to 95.9 mg/g and in Satureja hortensis L. ranged from 72.1 mg/g to 95.1 mg/g. In both species concentrations significantly decreased during the plant development. Flavonoids content in both species decreased during the vegetation. It was ranging in Satureja montana L. between 5.4 mg/g - 11.8 mg/g and in Satureja hortensis L. from 12.4 mg/g to 21.5 mg/g. Using high-performance liquid chromatography analysis several phenolic compounds were identified and quantitated: rutin (0.8-9mg/g), ferulic acid (1.9 to 7.4 mg/g), naringenin (0.11 to 0.45 mg/g) and kaempherol (3 0.7 to 10, 5 mg/g). Total radical scavenging activity in Satureja montana L. was the stronger- 66.6 mg/g - 106.5 mg/g than in Satureja hortensis L. (56.7 mg/g - 71.2 mg/g). Keywords: Satureja hortensis L., Satureja montana L., phenolic compounds, essential oils, radical scavenging activity, GC-MS, HPLC

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P-55 COMPARATIVE ANALYSIS OF RED AND WHITE WINES USING SPECTROPHOTOMETRIC AND CHROMATOGRAPHIC METHODS Raimonda Adomaitytė, Ieva Akuneca, Mantas Stankevičius, Audrius Maruška Departament of biochemistry and biotechnologies, Vytautas Magnus university, Kaunas

The objective of this study was to compare antioxidant activity, phenolic compounds and flavanoids content and to analyze volatile compounds in white and red wines. Red and white wines from various varieties (such as Cabernet Sauvingnon, Chardonnay, Cabernet Sauvingnon and Merlot, Cabernet Sauvingnon and Syrah, Chenin blanc, Chardonnay and Semillon, Pinot, Grigio and Chardonnay, Terret, Clairette and Maccaben) of the grapes were assayed for their antioxidant properties, total amount of flavanoids and total amount of phenolic compounds. Total amount of phenolic compounds of the samples was determined by Folin Ciocalteu method, antioxidant activities were evaluated using the DPPH· radical. Total amounts of phenolic compounds varied from 367.59 to 635.01 mg of rutin equivalents (RE) / 100 ml in red wines and from 55.21 to 88.42 mg RE / 100 mg in white wines. Total amount of flavanoids in red wine varied from 21.23 to 56.55 mg RE / 100 mg while in white wine 1.07 – 6.79 mg RE / 100 mg. Volatile compounds were analyzed by gas chromatography using solid phase microextraction (SPME) and direct headspace (HS) injection. There were detected 44 different volatile compounds in all samples. They provide grassy, herbaceous, fruity, floral odor. 16-24 volatile compounds were detected by SPME method for the sample preparation, 1-6 compounds by direct HS injection method for the sample preparation. Totally 44 different volatile compounds were detected in all samples. 2 compounds were detected which give bad smell to the wine: octanoate and octanoic acid – unpleasant rancid odor. Keywords: phenolic compounds, volatile compounds, wine, radical scavenging activity, GCMS, HS, SPME

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P-56 ANALYSIS OF DIFFERENT GEOGRAPHIC ORIGIN BEECOLLECTED POLLEN USING SPECTROPHOTOMETRIC AND CHROMATOGRAPHIC TECHNIQUES Geralda Ruočkuvienė1, Vilma Kaškonienė1, Olga Kornyšova1, Audrius Maruška1 1

Vytautas Magnus University, Faculty of Natural Sciences, Department of Biochemistry and Biotechnologies, Vileikos 8, Kaunas, LT-44404, Lithuania

The main aim of this study was to investigate and to identify biologically active compounds in bee-collected pollen from different geographic origin by means of spectrophotometric and chromatographic techniques. Bee pollen extractions were prepared by adding 20 ml of 80% methanol to 2 gram of raw material and 24 hour shaking in orbital shaker. The methanol extracts were filtered. Spectrophotometric methods were used to determine the total amount of phenolic compounds, flavonoids and antioxidant activity. The phenolic compounds content was determined using Folin–Ciocalteu method. It varied between 24.11 and 43.89 mg RE/g. The highest content of phenolic compounds was found in monofloral CHN_1 pollen 43.89 mg RE/g. The lowest content was found in polyfloral CHN_2 pollen - 24.11 mg RE/1 g. Similar tendency was observed for spectrophotometrically determined flavonoids content and total 2,2 – diphenyl – picrylhydrazyl (DPPH•) radical scavenging activity. Flavonoids content varied from 6.62 to 12.30 mg RE/g, free radical scavenging activity was in the range from 7.89 to 35.59 mg RE/g. Total amount of phenolic compounds showed strong correlation with antioxidant activity (correlation coefficient was 0.8507). For qualitative and quantitative analysis of bee pollen flavonoids a high performance liquid chromatography with electrochemical detection (HPLC-ED) was used. Four flavonoids were identified in all bee pollen samples: rutin, 2-hydroxycinnamic acid, naringenin and quercetin. Keywords: high performance liquid chromatography (HPLC), flavonoids, phenolic compounds, bee pollen, DPPH• radical scavenging activity

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P-57 SELF-GROWTH OF GOLD-POLYPYRROLE NANOCOMPOSITES Reda Kubiliūtė1, Justas Svirelis1, Lina Mikoliūnaitė1, Jaroslav Voronovič1, Anton Popov1, Almira Ramanavičienė1,2, Arūnas Ramanavičius1,3 1

Center of Nanotehnology and Material Science, Faculty of Chemistry, Vilnius University, Lithuania Laboratory of Immunoanalysis and Nanotechnology, Institute of Immunology of Vilnius University, Lithuania 3 Laboratory of NanoBioTechnology, Institute of Semiconductor Physics, State Research Institute Centre for Physical and Technological Sciences, Lithuania 2

Nanocomposites of conducting polymers and metal nanoparticles have received considerable attention due to the potential possibilities to create suitable materials for electrocatalysis, chemical sensors, and microelectronic devices [1]. In this work, nanoscaled gold-polypyrrole (Au-PPy) composites were prepared via in-situ polymerization of pyrrole using metal salt chloroauric acid as an oxidant. Preliminary experiments using HAuCl4 and gold nanoparticles (AuNPs) demonstrated a very fast self-growing of AuNPs. In this case, however, it is possible to prevent macroscopic growing by formation PPy layer on AuNPs surface. When this solution was treated with pyrrole, polymerization occurred, yielding PPy under concurrent formation of Au nanoparticles, as supported by UV-Vis measurements. For the next experiments reaction mixture consisting of AuNPs, HAuCl4 and pyrrole was used. The core shell particles consisting of AuNPs surrounded by a shell of conducting polymer might be expected to produce charge transfer at the interface due to the different chemical nature of the two materials. Also, the optical functions of both materials were drastically different from the corresponding bulk materials. After mixing all reaction components, the HAuCl4 solution containing AuNPs and pyrrole shows valence-to-conduction band transition at 750 nm. In addition, the absorption band of zerovalent AuNPs was observed at ca 570 nm after 2 days. This fact suggests that AuCl4- ion was reduced. The absorbance of AuNPs at 570 nm was blue shift with an increasing time and a new absorption band was observed at ca. 460 nm, and its intensity was increased with an increasing time. The absorption band at 460 nm could be assigned as π-π transition of PPy, but no absorption band due to charge carriers, i.e. polarons at 685 nm, was present. This means that the surface of the composite entirely consists of PPy molecules and the core of gold atoms, but PPy formed on AuNPs was not doped during the polymerization. In this system, we observed Au-PPy nanocomposites formation which induced AuNP-seeded growth in the presence of HAuCl4. This crystal growth was internally regulated by sizedependent activity decrease of AuNPs and PPy surface passivation. Aknowledgment: The work was supported by Research Council of Lithuania, Support to research of scientists and other researchers (Global Grant), Enzymes functionalized by polymers and biorecognition unit for selective treatment of target cells (NanoZim’s), Project Nr. VP1-3.1-ŠMM-07-K-02-042. References 1. Huang K, ZhangY, Han D, et al., 2006. One-step synthesis of 3D dendritic gold/polypyrrole nanocomposites via a self-assembly method. Nanotechnology, 17 (1): 283-288.

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P-58 HERBAL MEDICINES TO PREVENT NERVOUS SYSTEM DISEASES Laimutė Jonaitienė, Edita Babonienė, Ona Ragažinskienė Faculty of Medicine, Kauno kolegija / University of Applied Sciences, K. Petrausko g. 15, LT-44162 Kaunas, Lithuania

Recently, more and more people complain of increased nervousness, insomnia, or arising out of the depression. In an increasingly accelerating pace of life, makes many people treat themselves, and pharmacy staff (pharmacists and pharmacy assistants) is very important to provide service to the public, providing information on medicinal products they can offer a patient resident who does not have a prescription from a doctor. Nominated complaints are selected medicinal plant preparations, after hearing the good and considering all the relevant information so that the product is selected correctly. Pharmacists and pharmacy technicians, who are selling plant-based medical, must know all plants used in the composition of the preparation, also must know all them indications, contraindications and the side effects for the people nervous system. The object - herbal medicines used to prevent and treat diseases of the nervous system and the population approach to pharmacy technician services in a pharmacy. The aim: To collect information about herbal medicines used for nerve diseases, registered in the State of medicines and medicinal substances register, marketed in pharmacies in five geographic regions of the Lithuanian population and to identify approaches to pharmacy technician to inform the service about their healing properties. Keywords: herbal medicines, nervous system diseases, pharmacy, Lithuania WHO tradicional Medicine Strategy 2002 – 2005, Document WHO/EDM/TRM/2002.1, World Health Organization, Geneva.

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P-59 MEDICINAL RAW MATERIAL DRYING TECHNOLOGY AND ITS BIOLOGICALLY ACTIVE SUBSTANCES EVALUATION Algirdas Raila1, Egidijus Zvicevičius1, Aurelija Kemzūraitė1, Aušra Čiplienė1, Audrius Maruška3, Ona Ragažinskienė2, Olga Kornyšova3, Vilma Kaškonienė3, Violeta Bartkuvienė3, Kristina Bimbiraitė-Survilienė3, Mantas Stankevičius3, Tomas Drevinskas3 1

Laboratory of Biomass Treatment, Logistics and Solid Fuel Processes, Institute of Energy and Biotechnology Engineering, Aleksandras Stulginskis University, Studentų g. 11 LT-53361 Akademija, Kauno raj., Lithuania 2 Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Ž.E. Žilibero str. 6, LT- 46324 Kaunas, Lithuania 3 Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos str. 8, LT44404, Kaunas, Lithuania

In order to extend the variety of plant species and solve the problems of National genetic resources protection and herbal drugs industry development. Since 1924 the scientists of Kaunas Botanical Garden of Vytautas Magnus University have been implementing longlasting trials dedicated for introduction of medicinal and spice (aromatic) plants (MAP) species and varieties. The topics and tendencies of the research on medicinal herbs varied since that time quite a lot. Collections of medicinal plants are still an important constituent and object of the raw material drying technology and phytochemical scientific research process. During the long-lasting investigation of MAP in the collections the plant growth and vegetation were evaluated. Based on the results obtained the most perspective, biologically and ecologically valuable herbs were selected. In order to evaluate the relevance of these MAP for food and drug industries, the quantitative and qualitative analysis of raw material, investigation of biological and anatomical qualities were performed. Object of investigation – medicinal raw material – Hyssopi herba, drying technology and biologically active substances. Phytochemical analysis was performed using various chromatographic, capillary electrophoresis and spectrophotometric methods; solid phase microextraction, supercritical fluid extraction, maceration and solid phase extraction were used for sample preparation. While selecting the drying technology of Hyssopi herba and other medicinal and spice plants, it is essential to balance the airfl ow and its temperature in order to prevent the increase in the activity of micromycetes which could occur due to dampening of the raw material or prolonged drying process, and to avoid the possibilities of formation and accumulation of toxic compounds. It is particularly important when the drying agent is additionally heated because in cases of too low or too high heated airfl ow, it is more diffi cult to avoid condensation and sorption as well as formation of poorly drying zones. Keywords: hyssop, drying technology, biologically active substances. Raila A., Lugauskas A., Kemzūraitė A., Zvicevičius E., Ragažinskienė O., Railienė M. Different drying technologies and alternation of mycobiots in the raw material of Hyssopus Officinalis L // Annals of Agricultural And Environmental Medicine. ISSN 1232-1966. Vol. 16, No. 1 (2009), p. 93-101.: http://www.aaem.pl/pdf/AAEM.htm

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P-60 ANALYSIS OF PHYTOCHEMICAL COMPOSITION AND ANTIBACTERIAL ACTIVITY IN MEDICINAL PLANT MYRRHIS ODORATA L. Mickienė Ruta1, Friese Anika3, Rosler Uwe3, Maruška Audrius1, Ragažinskienė Ona2 1

Faculty of Natural Sciences, Department of Biochemistry and Biotechnologies, Vytautas Magnus University, Vileikos str. 8, LT- 44404 Kaunas, Lithuania 2 Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Ž.E. Žilibero 6, LT- 46324 Kaunas, Lithuania 3 Institute for Animal Hygiene and Environmental Health, Department of Veterinary Medicine, Free University Berlin. Robert-von-Ostertag-Strasse 7-13 14163 Berlin, Germany

The antibacterial activity of Myrrhis odorata L. was tested by the method of series dilutions, against different bacteria species. Investigated microorganisms were Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances mostly originating from healthy livestock. The extract from Myrrhis odorata L. showed antimicrobial activity against the investigated bacteria.A minimal inhibitory concentration (MIC) was applied as a method of measurement of the phytochemical activity. Preliminary results show that the MIC range between 8 – 13%.The most sensitive bacteria wasProteus vulgaris, an isolate without antibiotic resistance used for the official testing of disinfections withMIC of 8 %. The variations in the content and chemical composition of Myrrhis odorata L. have been grown in Lithuania were determined by gas chromatography/mass spectrometry (GC/MS). The highest content and diversity of compounds was determined during the full-flowering stage. The main components were anetole, methyleugenol, germacrene. Key words: Myrrhis odorata L., antimicrobial activity, herbal medicine, microorganisms

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P-61 INVESTIGATION OF BIOREMEDIATION OF CREOSOTE IMPREGNATED WOODEN RAILWAY SLEEPERS J. Mikašauskaitė1, N. Tiso1, V. Snieškienė2, A. Stankevičienė2, M.Stankevičius1, A. Maruška1, T. Drevinskas1, M. Kazlauskas1, V. Bartkuvienė1, O. Kornyšova1, O. Ragažinskienė2, D. Levišauskas3 1

Department of biochemistry and biotechnologies, Vytautas Magnus University, Vileikos 8, LT-44404, Kaunas, Lithuania 2 Kaunas Botanical Garden, Vytautas Magnus University, Z. E. Zilibero st. 6, Kaunas LT-46324 Kaunas, Lithuania 3 Process Control Department, Kaunas University of Technology Studentų St. 50, LT−51368 Kaunas, Lithuania

Railway sleepers impregnated with creosote or coal tar, contribute to the preservation of more than fifty years. Creosote, a distillation product of coal tar is one of the most widely used wood preservatives in the world. Used wooden sleepers are removed and stored in large quantities or being left in the environment. The Directive on Waste (75/442/EEC), (91/156/EEC), which establishes the EU waste policy, prohibits the uncontrolled waste disposal and requires to dispose waste without danger, noise and odors. The aim of this research is to determine the efficacy of selected basidiomycetes in the removing of polycyclic aromatic hydrocarbons (PAH) from the creosote-contaminated soil. Fungi Pleurotus ostreatus demonstrate positive interaction in creosote biodegradation. Quantity of creosote in treated wood sleepers can be estimated by measuring PAH content, using analytical methods like GC-MS, GC-FID, HPLC. Acknowledgement: This project is financed by EUSFA, grant No VP-3.1-ŠMM-10V-02_010 (BIOREM). Keywords: Pleurotus ostreatus, GC-MS, bioremediation, PAH, sleepers.

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P-62 QUANTITATIVE AND QUALITATIVE ANALYSIS OF POLYPHENOLIC AND VOLATILE COMPOUNDS IN PROPOLIS USING SPECTROPHOTOMETY, LIQUID CHROMATOGRAPHY AND GAS CHROMATOGRAPHY - MASS SPECTROMETRY METHODS Giedrė Blažukienė, Vilma Kaškonienė, Loreta Kubilienė, Audrius Maruška Department of biochemistry and biotechnologies, Vytautas Magnus University, Vileikos 8, LT-44404, Kaunas, Lithuania

Abstract Propolis is a natural product, consisting of plant resin and other biologically active, antioxidant compounds collected by honey bees. Therefore, propolis has attracted much attention in recent years as a useful natural raw material for pharmaceutical and cosmetic products. Furthermore, propolis is used in foods and beverages production, it can improve human health. The chemical composition of propolis depends on the collecting location, time of the year and plant source. Nine methanolic extracts of propolis of different botanical origin were analyzed using spectrophotometric methods, in order to determine total phenolic compounds, flavonoids and antioxidant activity. Five poliphenolic compounds were identified and their antioxiodant activity was assessed by means of high performance liquid chromatography with electrochemical detection (HPLC-ED). Twenty volatile compounds were identified by means of gas chromatography – mass spectrometry (GC-MS). The main volatile components indicate botanical origins of propolis. Keywords: phenolic compounds, volatile compounds, antioxidant activity, propolis, HPLC GC-MS

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P-63 ANALYSIS OF ESSENTIAL OILS AND PHENOLIC COMPOUNDS OF POTENTILLA FRUCTICOSA L. USING CHROMATOGRAPHIC AND SPECTROPHOTOMETRIC TECHNIQUES Šarūnas Paulaitis, Ieva Akuneca, Mantas Stankevičius, Ona Ragažinskienė, Audrius Maruška Department of biochemistry and biotechnologies, Vytautas Magnus University, Vileikos 8, LT-44404,

Abstract Otentilla fructicosa L. is one of the most common plants, planted in greeneries. Several tens of breed (decorative forms) differ in the form and height of the bush, colour shade of leaves, the size (from 2 to 5 cm) and colour of the flower: from snow white to various yellow shades. Potentilla fructicosa L. is spread in Europe, Asia and North America. The genesis of most of its breeds is unknown. In Lithuania until the end of this decade from woody cinquefoils only Potentilla fructicosa L. was planted. The goal of this work was to determine Potentilla fructicosa L. total amount of phenolic compounds, flavonoids and radical scavenging activity and the content of essential oils using spectrophotometric and chromatographic analysis methods. The experiment: methanolic Potentilla fructicosa L.extracts were prepared in this way: 0.5 g of fine-cut plant material extracted with 20 ml of 75 % methanol for 24 hours in the orbital shaker and filtrated. The total amounts of phenolic and flavonoids were determined, as well as antioxidant activity with different radicals by spectrophotometric method. HPLC was carried out to identify and quantitate phenolic compounds. Analysis of volatile compounds was carried out by gas chromatography coupled with mass spectrometry. Results: the highest amount of flavonoids in Potentilla fructicosa L. was determined in 2010 during the intensive blossoming (A1) 0.080 mgRE/g, and in 2012 (A1) 0.078 mgRE/g. The highest amount of phenolic compounds was determined in 2010, 2011 and 2012 samples collected at the beginning of blossoming period (Z1) 0.422 – 0.469 mgRE/g of dry plant material. The highest radical scavenging activity was observed in sample collected in 2012. Maximum value was determined during the flower bud development period (B) 0.806 mgRE/g and at the beginning of the blossoming period (Z1) 0.771 mgRE/g. Plants from more recent years are characterized by a smaller radical scavenging activity. After evaluation of radical scavenging activities with different radicals, it was determined that antioxidant activity with DPPH radical is different during various vegetation phases, the highest value was determined during the flower bud development period (B) 0.806 mgRE/g. Higher values were obtained with ABTS radical, maximum activities were determined during the intensive plant growing period (A1) 1.218 mgRE/g and during the flower bud development period (B) 1.215 mgRE/g. The highest walues were obtained using galvinoxyl radical, values obtained were almost double in comparison with DPPH radical. The highest value was determined during the flower bud development period (B) 1.723 mgRE/g. Investigating methanol extract radical scavenging activity dependence on colorimetric reaction time, it was determined that Potentilla fructicosa L. methanol extracts bound ABTS radical most effectively (remaining absorption after 120 min. was 0.07, i.e. 86%). Galvinoxyl radical’s remaining absorption after 120 min was 0.331, i.e. 33% and DPPH radical absorption was 0.363, i.e. 27%. It is important to highlight that plant extracts reacted fast enough with radicals (0-1min). After the performance of high performance liquid chromatography with electrochemical detection the main phenolic compound in all the collected samples was identified: gallic acid. The analysis of volatile compounds was performed. The main volatile compounds found in the samples using gas chromatography with mass spectrometry in all vegetation periods were hexanol, hexane, d-limonene and estragol. Keywords: Potentilla fructicosa L., phenolic compounds, essential oils, radical scavenging activity, GC-MS, HPLC

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P-64 ANTIOXIDANT ACTIVITY OF DIFFERENT BOTANICAL AND GEOGRAPHICAL ORIGIN HONEY Inga Šikštėnienė, Vilma Kaškonienė, Olga Kornyšova, Audrius Maruška Department of biochemistry and biotechnologies, Vytautas Magnus University, Vileikos 8, LT-44404,

Abstract Researchers are interested in natural antioxidant products, especially honey. In this work antioxidant activity of different geographical and botanical origin honey samples was investigated. Total phenolics, flavonoids, and radical-scavenging activity were determined using spectrophotometrical methods. Different geographical and botanical origin honey samples were extracted and preconcentrated using solid phase extraction (SPE) on C18 phase. Total phenolics were studied using the modified Folin-Ciocalteu method. Radical scavening activity was determined by the modified method of Blois. The highestt content of phenolics was found in buckwheat honey and the lowest in rape honey. The same tendency was observed for the flavonoids content. The ratio between total phenolic compounds and flavonoids was determined (ca. 5). The highest radical scavenging activity was determined for the darkest honey, and the lowest for the lightest. Other parameters, as vitamin C concentration, qualitative composition of the phenolic compounds and determination of volatile compounds were carried out. The data obtained will be presented and discussed in this report. Keywords: SPE, HPLC, flavonoids, phenolic compounds, honey, DPPH• radical scavenging activity

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L-65 EVALUATION OF PHENOLIC COMPOUNDS IN APPLE (MALUS DOMESTICA BORKH.) FRUITS Mindaugas Liaudanskas1, Raimondas Raudonis1, Valdas Jakštas1, Pranas Viškelis2, Darius Kviklys2, Juozas Lanauskas2, Nomeda Kviklienė2, Valdimaras Janulis1 1

Department of Pharmacognosy, Lithuanian University of Health Sciences, Mickevičiaus Str. 9, LT44307, Lithuania, [email protected] 2 Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, Kauno g. 30, LT54333 Babtai, Kaunas distr., Lithuania, [email protected]

Apples are a significant source of phenolic compounds which have a number of disease preventive effects [1]. The aim of this study was to develop a new method of investigating the profiles of phenolic compounds in different apple cultivars. Quantitative analysis of phenolic compounds from four apple cultivars (Auksis, Ligol, Lodel and Rajka) using reversed phase high performance liquid chromatography with photodiode array detection was carried out. Eight compounds from different phenolic groups such as flavonols, phenolic acids, flavan-3ols and dihidrochalcones were detected in the apples. The most abundant component in all apple cultivars was chlorogenic acid (the largest amount was in cv. Auksis 2903.83±39.21 µg/g). The fruit of all apple cultivars tested contained quercetin glycosides, namely hyperoside, rutin, avicularin and quercitrin. Hyperoside was the predominant form in all cultivars (the largest amount was in cv. Ligol 183.32±3.69 µg/g) except for cv. Rajka. The results obtained in this study give a new actual information about composition of phenolic compounds in tested apple cultivars. This method for determination of phenolic compounds in apple fruits will be used in our further investigations. Keywords: apple, phenolic compounds, HPLC, flavonoids Acknowledgement. This work was supported by a grant from the Research Council of Lithuania, No. SVE-02/2011. References: 1. Ćetković G., Čanadanović-Brunet J, Djilas S, Savatović S, Mandić A, Tumbas V (2008) Food Chem 109:340–347.

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P-66 INVESTIGATION OF GENETIC DIVERSITY OF IXODES RICINUS TICKS BASED ON MITOCHONDRIAL DNA AND MICROSATELLITE MARKERS Banaitytė Asta, Radzijevskaja Jana, Paulauskas Algimantas, Galdikaitė Eglė Departament of Biology, Vytautas Magnus University, Vileikos 8, LT-44414, Kaunas Lithuania

The application of molecular markers to the study of ticks has recently provided new insights into their population structures and phylogenetic relationships (Navajas and Fenton 2000). Use of microsatellites as genetic markers in population genetic studies has increased rapidly over the last years including several species of Ixodidae (Roed et al. 2006). Mitochondrial DNA (mtDNA) markers have also proved popular for studying of the systematic of ticks and their population structure. The aim of this study was to evaluate genetic diversity of I. ricinus ticks in Baltic countries using mtDNA and microsatellites markers. We screened three mitochondrial gene sequences 16S rRNA, cytb and control region sequences among 35 I.ricinus and ticks, collected from 16 locations: in Lithuania (4 locations), Latvia (4), Estonia (7) and Slovakia (1). Cytb gene was the most conservative gene that had only 4 haplotypes in Baltic countries, with control region sequences was detected 13 haplotypes and the most haplotypes (16 haplotypes) was detected in 16S rRNR gene sequences. We used four microsatellite loci for investigation of genetic diversity among 180 I.ricinus and ticks, collected from 16 locations in Lithuania (6 locations), Latvia (5) and Estonia (5). A total of 79 alleles were detected: the highest number of alleles per locus (25) was observed in samples from Latvia, followed in samples from Lithuania (20), and the lowest in samples from Estonia (18). The number of observed alleles between loci ranged from 5 to 10, and the observed heterozigosities between 0.2 and 0.8. Analysis of molecular variance (AMOVA) showed that genetic diversity of I. ricinus within sampling locations yielded 79%, and among locations 21% of the total genetic diversity. The PCA analysis not revealed distinct clustering for samples from different locations. Samples of different origin showed to a varying extent overlaps. Key words: ticks; Ixodus ricinus; microsatellite loci; mitochondrial gene References: 1. Navajas M., Fenton B. 2000. The application of molecular markers in the study of diversity in acarology: a review. Exp A ppl Acarol 24:751–774. 2. Roed K. H., Hasle G., Midthjell V., Skretting G., Leinaas P.H. 2006. Identification and characterization of 17 microsatellite primers for the tick, Ixodes ricinus, using enriched genomic libraries. 6, 1165–1167

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