ENVIRONMENTAL SCIENCE, ENGINEERING AND TECHNOLOGY
ADVANCED BIOLOGICALLY ACTIVE POLYFUNCTIONAL COMPOUNDS AND COMPOSITES: HEALTH, CULTURAL HERITAGE AND ENVIRONMENTAL PROTECTION
No part of this digital document may be reproduced, stored in a retrieval system or transmitted in any form or by any means. The publisher has taken reasonable care in the preparation of this digital document, but makes no expressed or implied warranty of any kind and assumes no responsibility for any errors or omissions. No liability is assumed for incidental or consequential damages in connection with or arising out of information contained herein. This digital document is sold with the clear understanding that the publisher is not engaged in rendering legal, medical or any other professional services.
ENVIRONMENTAL SCIENCE, ENGINEERING AND TECHNOLOGY Additional books in this series can be found on Nova’s website at: https://www.novapublishers.com/catalog/index.php?cPath=23_29&seriesp=Environmental+ Science%2C+Engineering+and+Technology
Additional e-books in this series can be found on Nova’s website at: https://www.novapublishers.com/catalog/index.php?cPath=23_29&seriespe=Environmental+ Science%2C+Engineering+and+Technology
ENVIRONMENTAL SCIENCE, ENGINEERING AND TECHNOLOGY
ADVANCED BIOLOGICALLY ACTIVE POLYFUNCTIONAL COMPOUNDS AND COMPOSITES: HEALTH, CULTURAL HERITAGE AND ENVIRONMENTAL PROTECTION
NODAR LEKISHVILI, GENNADY ZAIKOV AND
BOB HOWELL EDITORS
Nova Science Publishers, Inc. New York
Copyright © 2010 by Nova Science Publishers, Inc. All rights reserved. No part of this book may be reproduced, stored in a retrieval system or transmitted in any form or by any means: electronic, electrostatic, magnetic, tape, mechanical photocopying, recording or otherwise without the written permission of the Publisher. For permission to use material from this book please contact us: Telephone 631-231-7269; Fax 631-231-8175 Web Site: http://www.novapublishers.com NOTICE TO THE READER The Publisher has taken reasonable care in the preparation of this book, but makes no expressed or implied warranty of any kind and assumes no responsibility for any errors or omissions. No liability is assumed for incidental or consequential damages in connection with or arising out of information contained in this book. The Publisher shall not be liable for any special, consequential, or exemplary damages resulting, in whole or in part, from the readers’ use of, or reliance upon, this material. Any parts of this book based on government reports are so indicated and copyright is claimed for those parts to the extent applicable to compilations of such works. Independent verification should be sought for any data, advice or recommendations contained in this book. In addition, no responsibility is assumed by the publisher for any injury and/or damage to persons or property arising from any methods, products, instructions, ideas or otherwise contained in this publication. This publication is designed to provide accurate and authoritative information with regard to the subject matter covered herein. It is sold with the clear understanding that the Publisher is not engaged in rendering legal or any other professional services. If legal or any other expert assistance is required, the services of a competent person should be sought. FROM A DECLARATION OF PARTICIPANTS JOINTLY ADOPTED BY A COMMITTEE OF THE AMERICAN BAR ASSOCIATION AND A COMMITTEE OF PUBLISHERS. LIBRARY OF CONGRESS CATALOGING-IN-PUBLICATION DATA Available upon request ISBN: 978-1-61209-092-4 (eBook)
Published by Nova Science Publishers, Inc. New York
CONTENTS Preface
ix
Part 1.
Chemistry, Use and Molecular Modelling of Biologically Active Compounds
Chapter 1
Molecular Design and Reactivity of the 1-Hydroxycyclohexyl Hydroperoxide - Alk4nbr Complexes N. A. Turovskij, E. V. Raksha, E. N. Pasternak, I. A. Opeida, and G. E. Zaikov
Chapter 2
Chapter 3
Chapter 4
Chapter 5
Chapter 6
Chapter 7
Reception of Physiologically Active Substances by Plasmatic Membrane of Vegetable Cell Jaba Oniani, Vladimir Yurin, Ramaz Gakhokidze, Tea Mchedluri, Tamar Oniani and Lela Abadovskaya The Influence of Caffeine Analogues and Antagonists on the Ca2+Accumulation by Sarcoplasmic Reticulum Olga M. Alekseeva, Yuri A. Kim and Vladimir A. Rykov Biologically Active Multifunctional Adamantane-Containing Compounds Khatuna Barbakadze, Nodar Lekishvili, Zurab Pachulia, Giorgi Lekishvili, Badri Arziani, Iuri Sadaterashvili and Davit Zurabishvili Influence of Some Metal-Cations on the Molecular Organization of DNA N. Vasilieva-Vashakmadze, G. Lekishvili, R. Gakhokidze and P. Toidze State of Lipid Components of Soybean Flour Enzymatic Hydrolyzates during Storage L. N. Shishkina, E. V. Miloradova, E. A. Badichko and S. E. Traubenberg Participation of Aromatic Amines in the Maillard Reaction R. Kublashvili and D. Ugrekhelidze
1
11
25
35
61
69
81
vi Chapter 8
Contents Influence on Oxidation Processes Regulation Is the Reason for Biological Activity of the Ecdysteroid-Containing Compounds L. N. Shishkina, O. G. Shevchenko and N. G. Zagorskaya
Part 2.
Physics, Thermodynamics and Kinetics of Homogeneous and Heterogeneous Nanosystems
Chapter 9
Charge Transfer Mechanisms at Sam-Modified Electrodes Impact of Complex Environments Dimitri E. Khoshtariya, Tina D. Dolidze and Rudi van Eldik
Chapter 10
Chapter 11
Chapter 12
Chapter 13
Chapter 14
Chapter 15
Stability of Drug Delivery PLGA Nanoparticles: Calorimertric Approach Tamaz Mdzinarashvili, Mariam Khvedelidze, Tamar Partskhaladze, Mark Schneider, Ulrich F. Schaefer, Noha Nafee and Claus-Michael Lehr
87
103
129
The Tacticity Governed Stereomicrostructure in Poly(Methyl Methacrylate) (Pmma) as a Way to Explain Its Physical Properties N. Guarrotxena
147
Kinetics of the Fermentative Reaction of H2o2 Decomposition under the Action of Catalase in the Presence of Biosas for the Stationary State A. A. Turovsky, R. O. Khvorostetsky, L. I. Bazylyak and G. E. Zaikov
155
Thermodynamic and Kinetics of the Metalporphyrin – Base Reactions Tatyana N. lomova
167
Kinetics of the Fermentative Process in Stationary State for Sunflower-Seed Oil Hydrolysis by Lipase in the Presence of Biosas A. A. Turovsky, R. O. Khvorostetsky, L. I. Bazylyak and G. E. Zaikov Dynamic Trend of Energy Exchange Intensity in Brain Under Chronic Stress Nana Koshoridze
Part 3.
New Compounds for Medicine
Chapter 16
Synthesis and Transformation of New Anemia-Opposite Adamantane Derivatives of Ferrocene Oliko Lekashvili, Davit Zurabishvili, Levan Asatiani and Nodar Lekishvili
179
185
195
Contents Chapter 17
Pharmacological Premises of the Creation of New Antitumor Preparations of the Class of Nitrosoalkylurea J. A. Djamanbaev, Ch. Kamchybekova, J. A. Abdurashitova and G. E. Zaikov
Part 4.
Biofibers
Chapter 18
Grafting of Some Biofibres with Carboxylic Acids under Cold Plasma Conditions C. Vasile, M. Totolin and M. C. Tibirna
Chapter 19
Specific Properties of Some Biological Composite Materials N. Barbakadze, E. Gorb and S. Gorb
Part 5.
Compounds for Antibiocorrosive Covers and Protectors
Chapter 20
Antibiocorrosive Covers and Conservators Based on New Carbofunctional Oligosiloxanes and Biologically Active Compounds Nodar Lekishvili, Khatuna Barbakadze, David Zurabishvili, Tea Lobzhanidze, Shorena Samakashvili, Zurab Pachulia and Zurab Lomtatidze
Chapter 21
Synthesis, Biocide Properties and Structures of Some Arsonium Polyiodides for Antibiocorrosive Covers L. Arabuli, N. Lekishvili and M. Rusia
Part 6.
Environmental Chemistry
Chapter 22
Antimutageni and Anticytotoxic Activity of Bioenergoactivators Ramaz Gakhokidze and Amiran Pirtskhelani
Part 7.
Personally
Chapter 23
The Scientist who Has Outstripped His Time Revaz Skhiladze, Tengiz Tsivtsivadze and Bachana Pichkhaia
Index
vii
201
209 243
275
295
309
319 331
PREFACE During the last decade, researchers working within the field of biologically active compounds were attracted to finding new compounds, materials and methods that could be used toward the protection of human health and environment, along with the preservation of cultural heritage from various microorganisms, viruses and fungi. The actuality of this problem was stressed at the 41st congress of IUPAC devoted to these and related topics. Many actual aspects on creation of new compounds were discussed, materials and methods capable of more effective solving of the aforementioned problems. The topics that are discussed in this book encompass novel bioactive systems for human and environmental protection and preservation of cultural heritage from the microorganisms’ and fungi attacks. Also presented are the results of theoretical and experimental investigations conducted by the experienced scientists from different countries, along with the mechanistic comprehension of their performance under various experimental conditions. Chapter 1 - Chemically activated 1-hydroxycyclohexyl hydroperoxide decomposition in the presence of ammonium salts is proposed to proceed through the complexation stage. Complex structure and reactivity have been investigated by molecular modelling methods. Kinetics of the chemically activated hydroperoxide decomposition in the presence of quaternary ammonium salts (Et4NBr, Pr4NBr, Bu4NBr, and Hex4NBr) has been studied. The correlation between reactivity and structural characteristics of ammonium cations was found. Chapter 2 - Electrical phenomena and cytoplasm movement on an example of seaweed Characea have been briefly described. The results concerning cyclosis nascent mechanism in the cells of seaweed Characea and connection of cytoplasm movement velocity (CMV) with the difference of electrical potentials (DEP) of plasmatic membrane have been adduced. For specification of the appropriateness of interconnection of the cyclosis and DEP, the authors carried out the simultaneous measurement. A suggestion was made for a model in the frame of which it is possible to set up, in every separate case, a way to realize the observed effects by over regulation of the membrane potentials, i.e., finally by changing of ion penetrability of the membrane and in what—due to presence of other influences (for example, by influence on the difference intercellular processes). For ascertainment of the possibility of the presence on the plasmatic membrane of vegetable cell structures, similar to animal receptors and the interaction character of them, effectors, a detailed research had been conducted of acetylcholine acting and some biogenic amines on the velocity of cytoplasm and DEP of cell of seaweed Characea. Nitella flexilis.
x
Nodar Lekishvili, Gennady Zaikov and Bob Howell
Results show that a reaction of the cell on adrenomimetics, active in respect to α and βreceptors, is discerned by sign. Investigation of combined action of adrenomimetics and αtype blockaders (nor adrenalin-fenitron) and β-type (isadrin – dihydroergotoxin) showed the strongly pronounced antagonistic effect, realizing by concrete mechanism. There were adduced also other proofs allowing for the existence of interacting centers in the cells with molecules of catecholamines, similar to α and β-receptors of animal cells; while this resemblance is spread as on the structure organization, so it is on some functional features. Data about the presence of cAMF, cGMF, enzymes of their synthesis and catabolism (adenilatciclase, gaunilatciclase, phospodietherase) and protein-target – proteincinases, also G-protein, allowed authors of the work to make the conclusion about the specific interaction of acetylcholine and biogenic amines with the specialized structures of plasmatic membrane of vegetable cells and signal transmitting by trigger mechanisms on the G-protein with following participation of secondary intermediary messengers. Following from the above-stated consideration, the authors proposed the presumptive conclusions: There is revealed the phenomenon of receptor regulation of intercellular physiologic processes of vegetable cells. In the experiments with biogenic amines was stated the appropriateness in the direct correlation between alteration of CMV and DEP. The cells of Nitellaceae can be used in the model experiments for testing of biological active substances, inasmuch as there was revealed by them the property to react by constellation receptor system on the exogenous influence of low concentration of testing compounds. All those presuppositions show that in the plant cells is the presence of contractive proteins in view of actinomyozin complexes, otherwise who may answer the question which is open till today: What gives the first push to the cytoplasm particles to move constantly along the perimeter of the vegetable cell? Chapter 3 - This study investigates the actions of caffeine analogues and antagonists to the main Ca2+-depo in the rabbit skeletal muscle - sarcoplasmic reticulum. The efficiency of Ca2+-accumulation by vesicles of heavy fragmented sarcoplasmic reticulum was greatly changed by caffeine analogues and antagonist: cordiamin, camphor, bemegrid, 8methylcaffeine, teophilline, theobromine, midocalm. All tested substances have some similar characteristics at its molecular structure. The carbonyl oxygen and methyl groups are of a great importance for “caffeine effect” of these substances. Chapter 4 - New biologically active adamantane-containing anilides and nitroanilides have been synthesized and studied. By the semiempirical quantum-chemical method, AM1 effective charges on the atoms, bonds lengths and valence angles, enthalpies of formation of initial compounds and probable obtained products of the reaction of nitration of the anilides have been calculated. Based on quantum-chemical calculations and experimental data, the direction of the reaction of nitration has been established. To model physical properties of biologically active 16 anilides and nitroanilides, the authors studied quantitative “structureproperties” relationships (QSPR) on the basis of the experimental data. The authors used several sets of molecular descriptors. The dataset outliers were identified by using Principal Component Analysis (PCA). As a modeling technique, the authors applied Projections to Latent Structures (PLS). To ascertain the quality of models, the authors used cross-validation.
Preface
xi
Based on the author’s research, one concludes that the best models were acquired by the use of GETAWAY. The anthelminthic activity and activity towards various microorganisms of the obtained compounds have been studied. Based on preliminary investigations, it was established that the obtained compounds may be recommended as modifiers of anthelminthic preparations— phenacetyne, trinoine, diamphenetide, raphoxanide—also as a bioactive component for preparation: a) materials with antimycotic properties for prophylaxis and treatment of mycosis and dermatomycosis; b) protective covers stable to biocorrosion from action of some mycopathogenic microorganisms. Chapter 5 - This work deals with the problem of modeling of a possible mechanism of point mutations of DNA under the influence of Ni2+ ions. Two feasible schemes of interaction of Ni2+ with nucleosides are considered. The first scheme presents the formation of the planar complex of Ni2+-G-C, and the second shows the incorporation of Ni2+ between the two neighboring complementing pairs. The authors used the MOPAC package to compare the force constants and energies of intramolecular hydrogen bonds in the complexes and corresponding values in a free G-C pair. The comparison enabled us to make a conclusion implying that the formation of the Ni2+-G-C complex is accompanied by the weakening of a hydrogen bond, nearest the joining point of Ni2+. The incorporation of Ni2+ between the two neighboring complementing pairs of G-C causes the weakening of all the three pairs of hydrogen bonds, but to a lesser extent. It has been demonstrated that in the first case the probability of point mutations, the replacement of G-C by A-T increases, and the probability of divisions, the fallout of triplets of type GGХ or ХGG increases in another case. Chapter 6 - The influence of hydrolysis and centrifugation processes of soybean semifat flour on various indices of the lipid component and dynamics of changes in the composition and characteristics in hydrolyzates within three months of storage were studied. It was shown that processes of hydrolysis and centrifugation, and also storage, cause reliable changes of the physical and chemical characteristics and lipid composition in hydrolyzates. Chapter 7 - Some patterns of the relationship of the interaction between aromatic amines (o-, m- and p toluidin; o-, m- and p-amino phenol; o-, m- and p-amino benzoic acid) and aldoses (D-glucose, D-galactose, D-mannose, L-rhamnose, D-xylose, L-arabinose, Dmaltose, D-lactose) in the Maillard reaction are investigated. In the Maillard reaction, the reactivity of aniline, toluidines and amino phenols increases and the reactivity of amino benzoic acids decreases, with increase of рН of the reaction medium; in comparison with aldohexoses, aldopentoses participate in melanoidin formation more actively. Chapter 8 - The influence of serpisten and inokosterone on the phospholipids composition in liver and blood erythrocytes, intensity of lipid peroxidation in tissues (liver, spleen, blood plasma), catalase activity in liver and general peroxidase activity of white outbreed mice has been studied. A biological activity of ecdysteroid-containing compounds is shown to be associated with an influence on the parameters of the physicochemical regulatory system of lipid peroxidation (LPO). Possessing pronounced membrane-tropic properties due to alterations in the exchange of predominantly choline-containing fractions of phospholipids, ecdysteroid-containing preparations are capable of modifying a cell membrane phase state. A substantial dependence of a biological effect of the compounds on a dose, duration of their
xii
Nodar Lekishvili, Gennady Zaikov and Bob Howell
application as well as on an intensity of the LPO processes in the tissues and an animal’s sex require a more detailed research on the properties of the given ecdysteroids. Chapter 9 - Nanoscopic electrochemical devices composed of SAM-modified electrodes and redox probes (RP) of different kind (complex ions, proteins, organic molecules, etc.), were proven to be suitable systems for studying intrinsic electron transfer (ET) mechanisms and interplay between them. In the present review the authors consider the author’s recent results on the mechanistic studies of Au/SAM/RP nanoscale devices in which the RP were either redox-active protein cytochrome C (CytC) dissolved in aqueous solution, or the complex compound ferrocene (Fe(Cp)2]0/+, Cp=cyclopentadienyl) dissolved in a room temperature ionic liquid (RTIL), [bmim][NTf2]. The SAM composition was either [-S(CH2)n-OH] with n=2,3,4,6 and 11, or [-S-(CH2)n-CH3] with n=1,2,3,5,7,11 and 17, respectively. The modern electrochemical methodology including fast scan cyclic voltammetry and data processing techniques were applied to extract ET rate constants and an impact of the variation of ET distance, viscous additives (or temperature) and high pressure was determined, allowing for a rigorous discrimination of intrinsic ET mechanisms. In particular, at short electrode-reactant separations, n=2-3 for Au/SAM/CytC and n=1-3, for Au/SAM/Fe(Cp)2]0/+, the adiabatic mechanism of ET controlled by the viscosity-related relaxational modes of the RP environment, found to be operating. At larger electrode-reactant separations, n=6-11 for Au/SAM/CytC and n=7-11 for Au/SAM/Fe(Cp)2]0/+, the nonadiabatic ET mechanism can be observed manifested through the exponential decay of rate constant with the increase of n. At n=4 and n=5, the intermediate (mixture) regime of ET can be detected. Furthermore, for the case of n=17 for Au/SAM/Fe(Cp)2]0/+ in [bmim][NTf2], dynamical arrest (broken ergodicity) for ET has been demonstrated. In overall, together with other matching proceedings, the reviewed results, despite of essentially diverse nature of complex environments invoked within two different series of congruent systems (protein/aqueous solution versus RTIL) demonstrated common general patterns of the mechanism changeover, in a nice agreement with the predictions of a generalized theory of ET. Chapter 10 - The spherical PLGA nanoparticles (NP) calorimetric investigation is presented in this paper. Such nanoparticles is used for biological active substances (drugs) encapsulating inside of them with the purpose of medicine transferring into the cell. It is clear that without determination of particle stability it is impossible their practical usage. From calorimetric study of PLGA nanoparticles with PLA/PGA ratio 70:30 it was determined the entirety conditions of such particles and the temperature interval, where the particle destructions take place. It was unambiguously shown that for noncoated PLGA NP and for chitosancoated PLGA NP the stability temperature are equal to 370C and less than physiological temperature, which exclude their practical application. Also it was determined that hermiticity destroy temperature depends on heating rate. At the same time it was established that strongly alkaline and acid area (pH2 – pH9) do not destroy noncoated PLGA NP and chitosancoated PLGA NP what gives possibility for their using orally. Chapter 11 - Three industrial samples of Poly(methyl methacrylate) (PMMA), prepared under different conditions, have been extensively analyzed by means of 1H-NMR spectroscopy. Starting from the mm, rr and mrandrm triad contents, as given by the spectra, the type of tacticity statistics distribution has been deduced. Sample X appears to be completely Bernoullian, while samples Y and Z deviate somewhat from this behaviour
Preface
xiii
exhibiting a tiny trend towards Markovian statistics. The fraction of mmrm and rrrm pentads and that of pure heterotactic and atactic triad moieties has been calculated by assuming either a Markovian statistics for samples Y and Z or a Bernoullian statistics for all the samples. On the other hand, the fraction of the same pentads has been determined by deconvoluting the overall triad signals of the spectra into the corresponding pentad signals. An appreciably good agreement with the values obtained assuming Bernoullian statistics for all the samples appears evident. As a result, the evolution of every pentad content from sample X to Sample Z could be stated. Thus the samples prove to be appropriate models to study the relationship between any physical property and the stereomicrostructure of PMMA as was done previously for Poly(vinyl- chloride) (PVC) and Polypropylene (PP). Chapter 12 - It was investigated the fermentative stationary kinetics of hydrogen peroxide decomposition under the action of catalase in the presence of bioSAS. It were obtained the kinetic parameters of this process. It was shown, that the bioSAS have influence on the fermentative process, which can be explained by the change of the fermentative center activity or by the change of substrate concentration. It was determined that the temperature of a process has an insignificant influence on the value of kinetic parameters. Chapter 13 - The results of studying the axial coordination of tetraphenylporphyrin complexes of high-charged metal cations (AcO)CrTPP, O=Mo(OH)TPP and O=W(OH)TPP with molecular ligands (hydrogen sulfide, imidazole and pyridine) in toluene are discussed. The thermodynamic and the kinetic characteristics of reactions between metalloporphyrin and molecular ligand were obtained by the method of spectrophotometric titration and chemical kinetics. Correlations between the molecular ligand basicity and the molecular complex stability are discussed. Chapter 14 - The catalytic rate constants for the process in the presence of bioSAS by different concentrations have been obtained. It was shown, that the constants some increase at bioSAS concentration increasing up to their micelle−formation beginning. The temperature has a slight influence on the value of catalysis constants, that can be explained by practically zero activation energies and depend on activation entropy. Chapter 15 – The authors studied dynamic trend of changes in the activity of creatinу kinase, aldolase and succinatdehydrogenase in brain cells under 30-day long stress induced by isolation and violated diurnal cycle. It was shown that these enzymes heterogeneously responded to 30-day long stress. Particular sensitivity was recorded by succinatedehydrogenase that showed the decline of activity in various sections of the brain at 60-80% on average. Unlike succinatedehydrogenase, aldolase activity increased on the 10th day of stress and then declined. Similar results were seen in relation to phosphokinase activity. It was observed that the change in the activity of the enzymes in question was accompanied by quantitative changes in nitric oxide-NO. In accordance with experimental data the authors suggested that the main signal molecule causing changes in enzyme activity should be NO. Chapter 16 - Hydrazides are characterized with different pharmacological activity, amongst, compounds with inhibitory action of hydrophobia virus and human immunodeficiency virus. Therefore, the authors considered as perspective synthesis of adamantine containing hydrazides and anaemia opposite ferroocenes admixtures. FerroceneA(ferrocenyl-1-phenyl-dioxy-1,4-butin-2) hes significant antitumour and antibacterial properties. The derivatives of adamantine have broad pharmacological activity, low toxicity,
xiv
Nodar Lekishvili, Gennady Zaikov and Bob Howell
high membranotropic and antibacterial properties. The search new biologically active compounds, the authors have synthesized ferrocene- and adamantine-containing derivatives. Chapter 17 - Perspectives in the field of creation of highly effective anticancerogenic preparations have been evaluated. For their creation is offered a new regio-selective method of glycosylation of alkylurea in conditions of nucleophilic catalysis with some following nitrosing of glycosyl carbamides of the D- and L-rows. This method opens principally new possibilities for modification of compounds by means of glycosylamides bond leading to preparations possessing small toxicity and high selectivity. Chapter 18 - Physical and biochemical functionalisation of bast fibres are ways to improve thermo- and moisture regulation, anti-bacterial anti-allergies, hygiene, creating “smart” textile. Enhancing natural properties of vegetable fibres is an intermediary step in the obtaining of new products with special applications. The vegetable fibers are biodegradable, can be recycled, and in natural state are highly polar and hydrophilic. To improve the properties of the cellulosic fibers, the chemical and/or physico – chemical modifications were applied. The surface esterification of the natural polymer with acids can be carried out to obtain biodegradable materials, novel fibres with tailored functionalities for special applications. In this paper, starting from Spanish broom (Spartium junceum, syn. Genista juncea) fibers, under action of cold plasma discharges, and using different kinds of carboxylic acids, cellulose esters with short and long side chains have been synthesized. The new grafted polymers were characterized by FT – IR spectroscopy (ATR), XPS and SEM in order to assess the existence of incorporated functional groups. The thermal characterization of the obtained fibres reveals their particular behaviour. Chapter 19 - Miniaturisation of technical systems creates the need for today’s science and engineering to assess the mechanical properties of small volumes of material. A specific feature of the structure and the combination of the desirable properties across several different length scales are fascinating by the many examples in biology. Determining the extraordinary properties of natural materials at the nanometer scale is regarded as a very attractive target for materials science. Mechanical behavior of various biological materials such as insect and plant cuticles was studied by applying experimental approaches of material science in order to explain their structural design and working principles. Experiments were performed on the head articulation cuticle of the beetle designed for friction minimisation and on the wax covered plant surfaces adapted for attachment prevention. Both insect and plant cuticles are multifunctional composite materials and have a multilayered structure. Gula cuticle of the beetle Pachnoda marginata is a part of the head articulation, which is a micromechanical system similar to a ball bearing. The surfaces in this system operate in contact and must be optimised against wear and friction and provide high mobility within the joint. The measurements on the gula cuticle were performed in order to understand structure and mechanical behavior of the material working for friction minimizing. The wax layer on the plant surfaces is a barrier for the attachment system of insects. Antiattachment function could be caused by contamination of attachment pads of insect with the wax crystalloids. Increase in roughness due to location of the wax crystals on the plant surface causes decrease in the real contact area between the plant surface and attachment pads of the insect. These are two of the hypotheses why insects cannot walk on the plant surfaces structured with wax. Nanoindentations on different plant surfaces were performed in order to understand the deformation behavior of the wax layer. This study is believed to be one of the first for
Preface
xv
mechanically testing insect cuticle and the very first for wax-coated plant surfaces in native condition. Chapter 20 - New carbofunctional oligosiloxanes containing trifluorinepropyl and methacrylic groups at silicon atoms have been synthesized and studied. Biological active nitroanilides with spatial adamantane-containing groups and cadmium complex compounds based on them have been obtained. By using the data of IR and NMR spectral analyses the composition and the structure of synthesized compounds have been established. New composite materials of multifunctional application for individual and environmental protection, based on the obtained silicon-organic carbofunctional oligomers and complex compounds, have been created. It was shown that the created composites could be used as: a) protective covers (film materials and impregnating compositions) stable to biocorrosion; b) materials with antimycotic properties for prophylaxis and treatment of mycosis; c) biologically active polymer materials for protection of archaelogical and museum exhibits; d) for human protection during their contact with microorganisms. Preliminary investigations have shown that the synthesized compounds have also a real perspective to be utilized as accessible antioxidants towards the cancer. Chapter 21 - Quaternary arsonium triiodides [(Ph3AsCH2I]I3 and [(i-Bu)3AsCH2I]I3 have been synthesized and studied. The x-ray structures of [(Ph3AsCH2I]I3 and [(i-Bu)3AsCH2I]I3 have been determined. Crystals belong to the monoclinic (comp.1) system, space group P 21/n (No. 14) with a = 10.97 (1)Å, b=13.152 (1)Å, c=16.882 (1)Å , β=93.01 (1)o and to the triclinic system (comp.2), space group P-1 (No. 2) with a=8.413 (1)Å, b=9.109 (1)Å, c=15.876 (1)Å, α = 76.24 (1)o , β=75.60 (1)o, γ=75.26 (1)o. The structures were refined to an R value of 0.063 from 4082 (comp. I) and 0.091 from 4475 (comp.2) observed reflections. The As atom is coordinated tetrahedraly to the substituents and the anion has a linear structure. The synthesis of [R2(R')AsCH2I]I3 (where R= R' or R≠ R') are described. The possibility of the perspective application of synthesized compounds has been shown. Chapter 22 - Pollution of the environment is caused by human industrial and agricultural activity. It poses a harmful factor to the genetic apparatus of organisms to which are connected hereditary diseases, premature aging, cardiovascular problems, etc. Numerous experimental investigations have shown that many chemical factors are characterized by a mutagenic influence. Following these discoveries, the author’s laboratory elaborated many prophylactic measures, directed at prevention of genetic damage from the influence of harmful mutagenic agents on the organism. In the laboratory, the antimutagenic and anticytotoxic effect of bioenergoactivators (biorag, ragozan, ematon and ragil) on mice have been studied during mutation and cytotoxicity induced with the fertilizer ammonium nitrate and the pesticides (phosphamide, trichlorfon and celtan). The cytologic and genetic methods of investigation have been applied in the study. Experiments showed that tested bioenergoactivators exerted highly effective automutagenic and anticytotoxic action. Environmental pollution, which basically is caused by agricultural and industrial activity of human beings, comes back to them as factors harmful to organisms and their genetic apparatus, resulting in not only hereditary illness, malignant tumors and premature senescence, but also illnesses such as cardiovascular and digestive system disease, neural, allergy, and others.
xvi
Nodar Lekishvili, Gennady Zaikov and Bob Howell
In connection with an annual increase of chemical pollutants, science stands in the front of genetic danger. Among those problems whose solutions are first and foremost, the protection of organisms and their progenies from chemical mutagens existeing in the environment is the most actual one. Wide application of chemical preparations in medicine, agriculture, industry and everyday life, as well as the existence of a great amount of chemical pollutants in the soil, water and atmosphere, allow us to talk about a sharp alteration of the ecologic situation [1]. Chapter 23 - This article is dedicated to the 100 years anniversary of birth of Georgian prominent scientist, Professor Akaki Gakhokidze. He is one of outstanding representatives of Georgian chemist’s school. High theoretical preparation, mastery of experiment conduction, unusual scientific flair and intuition allow him to leave the great and light footstep for the posterity on the way of scientific research and pedagogical activity. Fundamental investigation of A.Gakhokidze won the international recognition. His woks was published and broadly considered in the special literature, monographs and manuals of chemistry. There are created the specific paragraphs as “Synthesis of Gakhokidze”, “method of DanilowGakhokidze” etc.
PART 1. CHEMISTRY, USE AND MOLECULAR MODELLING OF BIOLOGICALLY ACTIVE COMPOUNDS
In: Advanced Biologically Active Polyfunctional Compounds… ISBN 978-1-60876-114-2 © 2010 Nova Science Publishers, Inc. Ed: N. Lekishvili, G. Zaikov and B. Howell
Chapter 1
MOLECULAR DESIGN AND REACTIVITY OF THE 1-HYDROXYCYCLOHEXYL HYDROPEROXIDE ALK4NBR COMPLEXES N.A. Turovskij11, E.V. Raksha1, E.N. Pasternak1, I.A. Opeida2, And G.E. Zaikov23 1 Donetsk National University, Donetsk, Universitetskaya str. 24, 83055, Ukraine; Ukrainian-American Laboratory of Computational Chemistry, Kharkov, Ukraine, Jackson, USA
2 Institute of Physico-Organic and Coal Chemistry of the National Academy of Sciences of Ukraine, R. Luxemburg 70, 83114 Donetsk, Ukraine 3 Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygina, 117334 Moscow, Russian Federation
ABSTRACT Chemically activated 1-hydroxycyclohexyl hydroperoxide decomposition in the presence of ammonium salts is proposed to proceed through the complexation stage. Complex structure and reactivity have been investigated by molecular modelling methods. Kinetics of the chemically activated hydroperoxide decomposition in the presence of quaternary ammonium salts (Et4NBr, Pr4NBr, Bu4NBr, and Hex4NBr) has been studied. The correlation between reactivity and structural characteristics of ammonium cations was found.
1
E-mail:
[email protected]. 2 E-mail:
[email protected].
2
N. A. Turovskij, E. V. Raksha, E. N. Pasternak et al.
INTRODUCTION Peroxides and hydroperoxides are widely used as initiators as well as components of binary initiating systems in the processes of vinyl monomers polymerization, polymers modification [1-3], oxidation of organic substances by molecular oxygen [4, 5]. Initiating systems on the base of quaternary ammonium bromides were found to be the most effective in the case of the liquid phase oxidation of isopropyl benzene by O2 [5]. 1-hydroxycyclohexyl hydroperoxide - Et4NBr system initiates the reaction of liquid phase isopropyl benzene oxidation at 308-340 K [6] while the hydrocarbon oxidation in the presence of only one component of the binary system was not occurred. The present work presents the study of 1hydroxycyclohexyl hydroperoxide decomposition activated by the tetraalkylammonium bromides to investigate the role of ammonium salt cation in the process of chemical activation of peroxide bond.
EXPERIMENTAL 1-hydroxycyclohexyl hydroperoxide has been prepared from cyclohexanone and H2O2 in anhydrous ether according with [7]. Tetraalkylammonium bromides (Et4NBr, Pr4NBr, and Bu4NBr) were recrystallized from the saturated acetonitrile solution by addition of diethyl ether excess. The solvent (acetonitrile) purity was controlled by electrical conductivity value, which was within (8.5 ± 0.2)·10-6 Оm-1 sm-1 at 303 K. Reactions were carried out in the glasssoldered ampoules in argon atmosphere. Hydroperoxide kinetic concentration was controlled by the iodometric titration with potentiometric fixation of the equivalent point.
CALCULATION METHODS Quantum chemical calculations of hydroperoxides molecules and corresponding radicals were carried out by AM1 semiempirical method implemented in MOPAC package [8]. The RHF method was applied to the calculation of the wave function. Optimization of structure parameters of hydroperoxide and hydroperoxide complexes was carried out by Eigenvector Following procedure. The molecular geometry parameters were calculated with boundary gradient norm 0.01. Solvent effect was considered in COSMO approximation [9].
RESULTS AND DISCUSSION It was shown recently that cyclohexanone peroxides decomposition in the presence of Et4NBr proceeds with lower activation barrier as compared with thermolysis. This fact can be explained by the complexation in the studied system [6]. We assume that the activation extent of the peroxide bond in the complex correlates with hydroperoxide reactivity in the reaction of radical decomposition. Thus we investigated the molecular design and reactivity of the complexes in the system 1-hydroxycyclohexyl hydroperoxide – Alk4NBr.
Molecular Design and Reactivity…
3
MOLECULAR DESIGN OF THE 1-HYDROXYCYCLOHEXYL HYDROPEROXIDE - ALK4NBR COMPLEX To obtain the structure information the molecular modelling of the ROOH-Alk4NBr ionmolecular complexes has been carried out for the case of Et4NBr. In contrast to the aralkyl hydroperoxides (like isopropylbenzene hydroperoxide) the molecule of 1-hydroxycyclohexyl hydroperoxide does not contain the aromatic ring but there is the hydroxide group OH in their structure that also can participate in the intermolecular hydrogen bonds formation. We assume the model of complex formation with combined action of cation and anion (Figure 1) such as previously proposed model of substrate separated ion pair (SubSIP) [10, 11]. Formation of such type associate is accompanied by considerable conformation changes of the hydroperoxide fragment. Association between hydroperoxide molecule and the salt ions occurs by the intermolecular hydrogen bonds formation. It is confirmed by charge increasing on the hydrogen atoms of the corresponding bonds C-H3, O-H1, and O-H2 and elongation of these bonds from 1.11 Å to 1.17 Å and from 0.96 Å to 1.08 Å correspondingly. Calculations in COSMO approximation show that the electron density transfer from bromideanion to the hydroperoxide fragment is lower in the case of solvent effect taken into account (Table 1). Stage determining the rate of the hydroperoxide decomposition is that of O-O bond cleavage. Thus the reaction activation energy will be determined by the energy of peroxide bond homolytic decomposition. Some possible elementary stages of the complex decomposition with the free radical formation are listed in Table 2. It can be seen from the Table 2 that the electrone transfer from the bromide ion to the peroxide bond is improbable in this system because it is needed more energy inputs as compared with homolysis. We assume that SubSIP decomposes with free radicals formation due to homolytical rupture of the peroxide bond. On the Figure 1 the enthalpies of complexation are shown. In the case of SubSIP we assume that the complex is formed according to the exchange mechanism (1) while possibility of the threemolecular reaction (2) is low. Solubility of the Et4NBr in acetonitrile is sufficiently high, and reactivity of the ions and ion pairs in peroxide bond activation is similar. The threemolecular reaction (2) determines the second reaction order on salt while this value changes from 1 (at small salt concentrations) to 0 (at salt abundance) in experimental conditions. Thus the reaction (1) is rather preferable. Table 1. Electron and stereochemical characteristics of COOH-fragment in ROOH – Et4NBr complexes parameters qOα, e qOβ, e qH1, e qH2, e qH3, e ∠COαOβH qBr-
ROOH - 0.18 - 0.20 0.20 0.22 0.10 90.8 -
Et4N+…ROOH…Br- 0.24 - 0.26 0.25 0.27 0.17 33.4 - 0.81
ROOH…Br- 0.18 - 0.29 0.25 0.27 0.11 68.3 - 0.85
ROOH… - 0.20 - 0.23 0.22 0.04 0.10 77.5 -
4
N. A. Turovskij, E. V. Raksha, E. N. Pasternak et al. Table 2. Heats of some elementary stages of ROOH – Et4NBr complex decomposition
ROOH → RO· + ·OH Et4N+…ROOH…Br− → RO· + ·OH + Et4N+ + BrEt4N+…ROOH…Br− → RO· + ·OH + Et4NBr Et4N+…ROOH…Br− → RO- + ·OH + Et4N+ + Br· Et4N+…ROOH…Br− → RO· + OH- + Et4N+ + Br· ROOH…Et4N+ → RO· + ·OH + Et4N+
ΔH0, kJ mol-1 gas phase 148 481 65 615 435 192
acetonitrile 151 239 108 354 249 179
ROOH…Br- → RO· + ·OH + BrROOH…Br- → RO- + ·OH + Br· ROOH…Br- → RO· + OH- + Br·
242 188 376
208 219 323
Reaction
Figure 1. Complexes of 1-hydroxycyclohexyl hydroperoxide bromide.
Molecular Design and Reactivity…
5
Et4N+…Solv…Br− + ROOH → Et4N+…ROOH…Br− + Solv (ΔHr(2) = - 53.7 kJ·mol-1)
(1)
Et4N+ + ROOH + Br− → Et4N+…ROOH…Br− (ΔHr(3) = - 88.1 kJ·mol-1)
(2)
Corresponding models like Et4N+…ROOH…Br− were obtained for tetraalkylammonium salts (Pr4NBr, Bu4NBr, и Hex4NBr). Changes in peroxide bond energy (i.e. total of electronic and nuclear energies of two-center term O-O atom pair) can be suggested as extent of the peroxide bond activation. Stable peroxide structure has the lowest O-O bond energy (-12.08 eV for free hydroperoxide) and less stable (or activated) peroxide conformation has the highest one (-11.78 eV for complex Et4N+…ROOH…Br−). Difference between energy of stable and activated peroxide structure increases in the catalyst row: Hex4NBr, Bu4NBr, Pr4NBr, and Et4NBr indicating weakening of the O-O bond in the ROOH-Alk4NBr complex. From this point of view the O-O bond activation in the case of Et4N+…ROOH…Br− is the highest and in complex with Hex4NBr is the lowest. This suggestion is in correspondence with experimental data.
KINETIC FEATURES OF THE ACTIVATED HYDROPEROXIDE DECOMPOSITION Reaction of 1-hydroxycyclohexyl hydroperoxide (ROOH) decomposition activated by the tetraalkylammonium bromides (Alk4NBr) has been investigated at 323 – 353 K under conditions of ammonium salts abundance ([ROOH]0 = 5·10-3 mol·dm-3, [Alk4NBr]0 = 2.5·102 – 1·10-1 mol·dm-3). Kinetics of ROOH activated decomposition on these conditions could be described by the first order reaction proceedings low. The reaction was carried out up to 50 % hydroperoxide conversion and the products did not effect on the reaction proceeding as kinetic curves anamorphouses are linear in the corresponding first order coordinates. The effective rate constant (kef, seс-1) was found to be independent from the hydroperoxide initial concentration within [ROOH]0 = 2.5·10-3 – 1·10-2 mol·dm-3 and constant amount of Alk4NBr (5·10-2 mol·dm-3). Nonlinear character of the dependence of reaction rate effective constants from the ammonium salt initial concentration (Figure 2) in the present conditions points out onto the occurrence of complexation stage between the ROOH and Alk4NBr. These facts are in an agreement with reaction scheme for the isopropyl benzene hydroperoxide activated decomposition recently proposed [12]. Thermolysis contribution into the total reaction rate the ROOH activated decomposition was found to be negligibly small because the thermal decomposition rate constant [6] was two orders lower then corresponding kef values. Some experiments were carried out under conditions of ROOH excess as compared with salts concentration ([Alk4NBr]0 = 5·10-3 mol·dm-3, [ROOH]0 = 5·10-2 mol·dm-3) at 323 K. Even at present concentration ratio of ROOH and Alk4NBr the rate of hydroperoxide activated decomposition is several orders higher then ROOH thermolysis rate. The Brconcentration during reaction proceeding is unaltered. These facts point out onto catalytic character of Alk4NBr action in the process. Catalytic scheme of the hydroperoxide decomposition is supported also in works [13, 14] in which it was noted that O-O bond
6
N. A. Turovskij, E. V. Raksha, E. N. Pasternak et al.
cleavage in the hydroperoxide – catalyst complex proceeds homolytically; catalyst is not consumed and deactivated in the system.
Figure 2. Dependence of kef from the catalyst initial concentration in the direct (a) and inverse (b) coordinates. [ROOH]0 = 5.0·10-3 mol·dm-3, 333 K, 1 – Hex4N+, 2 – Bu4N+, 3 – Pr4N+, 4 – Et4N+.
The kinetic scheme is proposed for the chemically activated ROOH decomposition. It includes the complexation stage between ROOH and salts ions as well as stage of the complex-bonded hydroperoxide decomposition with catalyst regeneration: kef dependence on the salt concentration can be expressed by the relationship (3):
1 1 1 = + k ef k d K C [ Alk 4 NBr ] k d
(3)
The dependence of kef on Alk4NBr concentration is linear in double inverse coordinates (Figure 2). It is in the agreement with the proposed kinetic scheme. The values of rate constants of complex-bonded peroxide decomposition (kd) obtained for the investigated salts decrease in the following order: Et4N+ > Pr4N+ > Bu4N+ > Hex4N+. The values of equilibrium constants of complexation between ROOH and Alk4NBr ions change similarly (Table 3). Isokinetic relationship between complexation parameters in the system leads to the insignificant changes in free energy of complexation for different alkyl substituent in ammonium cation. Considering the intermolecular bonds energy the strongest complex is formed between hydroperoxide and Et4NBr, the weakest - in the case of Hex4NBr (see the corresponding ΔHcom values in Table 3). The reactivity of complex-bonded ROOH also decreases with increasing of alkyl substituent size. Isokinetic relationship for the obtained activation parameters points out upon the unified mechanism of ROOH - Alk4NBr complex decomposition. Changes of the peroxide bond activation in the complex in the case of different ammonium cations point out the role of steric factor at the stage of complex formation as well as at the formation of their decomposition transition state. In the simplest case the own
Molecular Design and Reactivity…
7
volume of the investigated cations could describe the steric effect. A good correlation between activation parameters of the complex decomposition and calculated values of Vander-Waals volumes of cations has been obtained (Figure 3). VVDW values for the tetraalkylammonium cations were calculated in HyperChem package. Calculated values are in agreement with corresponding own volumes of cations listed in [15]. Linear relationship has been observed between the complex heat of formation - ΔH0f and experimental activation parameters - ΔH≠ (Figure 4). Table 3. Kinetic parameters of 1-hydroxycyclohexyl hydroperoxide decomposition in the presence of thetraalkylammonium bromides parameters kd·104, sec-1
KC, dm3mol-1 Ea, kJmol-1 lgA, (A, c-1) ΔHcom, kJmol-1 ΔScom, Jmol-1K-1
T, K 333 343 353 333 343 353 333353
ROOH + Et4NBr 1.14 ± 0.04 2.5 ± 0.1 5.1 ± 0.2 36 ± 2 29 ± 2 23 ± 1 73 ± 1 7.5 ± 0.6 -20 ± 1 -30 ± 4
ROOH + Pr4NBr 0.8 ± 0.1 1.80 ± 0.09 4.20 ± 0.09 28 ± 3 24 ± 1 20 ± 1 81 ± 2 8.6 ± 0.3 -16 ± 2 -21 ± 5
ROOH + Bu4NBr 0.59 ± 0.06 1.51 ± 0.04 3.48 ± 0.08 23 ± 2 20 ± 1 18 ± 2 87 ± 3 9.4 ± 0.2 -12 ± 2 -10 ± 4
ROOH + Hex4NBr 0.34 ± 0.01 0.97 ± 0.02 2.40 ± 0.06 18 ± 3 16 ± 2 15 ± 2 96 ± 2 10.5 ± 0.3 -9 ±1 -3 ± 3
Figure 3. Relationship between activation parameters of the complex decomposition and Van-derWaals volume of the tetraalkylammonium cations.
8
N. A. Turovskij, E. V. Raksha, E. N. Pasternak et al.
Figure 4. Relationship between the complex heat of formation (ΔH0f) and experimental activation parameters (ΔH≠).
Obtained experimental facts have shown that the salt cation participates both in complexation stage and in stage of complex ROOH - Alk4NBr decomposition. Ammonium cation has the regulating action upon the catalytic reactivity of the halide-anion in the reaction of catalytic decomposition of the 1-hydroxycyclohexyl hydroperoxide in the presence of Alk4NBr. Thus the cation structure influences on the reactivity of the hydroperoxide complex and on the extent of peroxide bond activation. The molecular modeling of the hydroperoxidecatalyst reactive complex can be used to preliminarily predict the reactivity of the system.
CONCLUSIONS Summarizing, obtained experimental facts have shown that the chemical activation of the peroxide bond is observed in the presence of Alk4NBr. The kinetic parameters of the hydroperoxide catalytic decomposition have been obtained for the Et4NBr, Pr4NBr, Bu4NBr, and Hex4NBr. Catalysis of the1-hydroxycyclohexyl hydroperoxide decomposition has shown to occur through accompanied action of the ammonium salt cation and anion.
REFERENCES [1] [2] [3]
E.T. Denisov, T.G. Denisova, T.S. Pokidova. Handbook of Free Radical Initiators. John Wiley and Sons Inc.: Hoboken, New Jersey, 2003, 879 p. N.A. Turovskij, I.O. Opeida, O.V. Kush, E.L. Baranovskij. J. Appl. Chem. 2004, 77, 1887. C.J. Perez, E.M. Valles, M.D. Failla. Polymer, 2005, 46, 725,
Molecular Design and Reactivity… [4] [5] [6] [7] [8] [9] [10]
[11] [12] [13] [14] [15]
9
I.A. Opeida, N.M. Zalevskaya, E.N. Turovskaya. Kinetika i Kataliz, 2004, 45, 776, I.A. Opeida, N.M. Zalevskaya, E.N. Turovskaya, U.I. Sobka, Petroleum Chemistry, 2002, 42, 6. 460. M.A. Turovskyj, I.A. Opeida, E.N. Turovskaya, O.V. Raksha, N.O. Kuznetsova, G.E. Zaikov. Oxid. Commun., 2006, 29, 249. N.A. Milas, S.A. Harris, P.C. Panagiotacos. J.Am.Chem.Soc., 1939, 61, 9, 2430. J.J.P. Stewart. MOPAC 2000.00 Manual; Fujitsu Limited: Tokyo: Japan, 1999. A. Klamt. J.Chem. Soc. Perkin Trans., 1993, 2, 799. M.A. Тurovskyj, I.O. Оpeida O.M. Turovskaya, O.V. Raksha, N.O. Kuznetsova, and G.E. Zaikov. Order and Disorder in Polymer Reactivity. Howell New York: Nova Scince Publishers, Inc. 2006, pp. 37-51. N.A. Turovskij, S.Yu. Tselinskij, Yu.E. Shapiro, A.R. Kal’uskij. Teor. Eksp. Khim., 1992, 28, 4. 320. M.A. Turovskyj, A.M. Nikolayevskyj, I.A. Opeida, V.N. Shufletuk. Ukrainian Chem. Bull., 2000, 8, 151. L.M. Pisarenko, O.T. Kasaikina. Russ Chem Bull, 2002, 3, 419. L.M. Pisarenko, V.G. Kondratovich, O.T. Kasaikina. Russ Chem Bull, 2004,10, 2110. Y.Marcus. Ion salvation. Chichester etc.: Wiley. 1985, 306 p.
In: Advanced Biologically Active Polyfunctional Compounds… ISBN 978-1-60876-114-2 © 2010 Nova Science Publishers, Inc. Ed: N. Lekishvili, G. Zaikov and B. Howell
Chapter 2
RECEPTION OF PHYSIOLOGICALLY ACTIVE SUBSTANCES BY PLASMATIC MEMBRANE OF VEGETABLE CELL Jaba Oniani1, Vladimir Yurin2, Ramaz Gakhokidze11, Tea Mchedluri3, Tamar Oniani1, and Lela Abadovskaya1 1
Iv. Jvakhishvili Tbilisi State University, Georgia 2 Minsk State University, Belarus 3 Telavi State University, Georgia
ABSTRACT Electrical phenomena and cytoplasm movement on an example of seaweed Characea have been briefly described. The results concerning cyclosis nascent mechanism in the cells of seaweed Characea and connection of cytoplasm movement velocity (CMV) with the difference of electrical potentials (DEP) of plasmatic membrane have been adduced. For specification of the appropriateness of interconnection of the cyclosis and DEP, we carried out the simultaneous measurement. A suggestion was made for a model in the frame of which it is possible to set up, in every separate case, a way to realize the observed effects by over regulation of the membrane potentials, i.e., finally by changing of ion penetrability of the membrane and in what—due to presence of other influences (for example, by influence on the difference intercellular processes). For ascertainment of the possibility of the presence on the plasmatic membrane of vegetable cell structures, similar to animal receptors and the interaction character of them, effectors, a detailed research had been conducted of acetylcholine acting and some biogenic amines on the velocity of cytoplasm and DEP of cell of seaweed Characea. Nitella flexilis. Results show that a reaction of the cell on adrenomimetics, active in respect to α and β-receptors, is discerned by sign. Investigation of combined action of adrenomimetics and α-type blockaders (nor adrenalin-fenitron) and β-type (isadrin – 1 E-mail:
[email protected].
12
Jaba Oniani, Vladimir Yurin, Ramaz Gakhokidze et al. dihydroergotoxin) showed the strongly pronounced antagonistic effect, realizing by concrete mechanism. There were adduced also other proofs allowing for the existence of interacting centers in the cells with molecules of catecholamines, similar to α and βreceptors of animal cells; while this resemblance is spread as on the structure organization, so it is on some functional features. Data about the presence of cAMF, cGMF, enzymes of their synthesis and catabolism (adenilatciclase, gaunilatciclase, phospodietherase) and protein-target – proteincinases, also G-protein, allowed authors of the work to make the conclusion about the specific interaction of acetylcholine and biogenic amines with the specialized structures of plasmatic membrane of vegetable cells and signal transmitting by trigger mechanisms on the G-protein with following participation of secondary intermediary messengers. Following from the above-stated consideration, we proposed the presumptive conclusions: 1. 2. 3.
There is revealed the phenomenon of receptor regulation of intercellular physiologic processes of vegetable cells. In the experiments with biogenic amines was stated the appropriateness in the direct correlation between alteration of CMV and DEP. The cells of Nitellaceae can be used in the model experiments for testing of biological active substances, inasmuch as there was revealed by them the property to react by constellation receptor system on the exogenous influence of low concentration of testing compounds.
All those presuppositions show that in the plant cells is the presence of contractive proteins in view of actinomyozin complexes, otherwise who may answer the question which is open till today: What gives the first push to the cytoplasm particles to move constantly along the perimeter of the vegetable cell?
As long ago as XIX century by known physiologist of animals Claude Bernard, the idea had been stated by what the irritability is one of the main properties of living organisms and for which the common mechanisms of perception and quick reaction on the outer influence are inherent. But the mechanisms of irritability, including the perception of outer stimulus, information on its transmission and in return reaction had been studied in the XX century [13]. Plasmatic membrane is the primary target of influence of exogenous factors, realizing the perception of their influence and transmission of the signal into the cell. Let us consider the approach developed by us of description of the physiological active substances reception with the structure elements of the plasmatic membrane of the vegetable cell on the base of analysis of regularity of an alteration of electrical characteristics of cyclosis speed. With this purpose, we will characterize phenomena and movement of cytoplasm on the example of seaweed Characea. Ability to generate electrical potential is one of the universal properties of the living system which plays an important role in their viability. It is known that bioelectrical potential today is subdivided on the two basic types: potential of immovability (PI) and potential of excitation (PE). PI is the difference of electrical potential (DEP) in the condition of immovability, PI-alteration of PE at the irritation [4-6]. PI or membrane potential is the main electrical characteristic of the plant cell and corresponding to it is the condition at physiologic
Reception of Physiologically Active Substances by Plasmatic Membrane…
13
immovability when the metabolism is in the balance condition. Disturbance of normal physiologic conditions of the cell inevitably leads to the change of PI value. On the membrane of any compartment of a living cell, there exists DEP, the absence of which would be amazing. It would mean the absolute equality of electrolyte concentration in all the cells, organs, outer solutions or complete coincidence membrane penetration value to all the cations and anions. Some examples of membrane potential value of the plant cell is shown in the table. On the base of PI all types of electro physiologic reactions of the cell, in particular excitation potentials, are formed. In the plants, the excitation potentials are represented by two types: potential of action (PA) and potential of variability (PV) [6,7]. PA is the temporary alternation of DEP between the excited section and the section being in the condition of immobility. It appears due to irritation and is spread as excitation waves. PA appearing as a result of the definite irritation force of the plant can be divided on the spreading—i.e. transmitting signal from cell to cell in the limits of the organ or tissue—and the local one, transmitting only in the limits of irritating cells. Table. Membrane potential of the plant cells (Klarkson D. 1978; Lutge Y., Khiginbotam H 1984) Species Tissue Crustof of root Pisum sativum Epicotel Caleoptil Avena sativa Root Zea mays Root Vicia sativa Root Nitella Internodal cell Nitellopsis obtusa Internodal cell Halicystis Cell Valonia Cell
Value of DEP, mV –110 –119 –102 – –109 –71 – –84 –76 – –104 –130 –120 – –150 –200 – –250 –80 +17
Elaborations of corresponding techniques and the interest of investigators in the electrical process in the plants allowed researchers, in the beginning of the past century, to register PA at their irritation. It was that the membrane potential of many vegetable cells, like those of animal origination, at the irritation is displaced to the side of depolarization. At the definite value of membrane potential displacement to the positive side till some critical level there is observed PA. Volume of displacement of membrane potential from initial level to critical one is called the threshold potential. The lower the threshold potential, the higher is the cell irritability. In comparison with animal objects PA, the potential of activity of vegetable cells is developed slowly. Another distinguished feature is that the PA of plant cells is developed on the two membranes—plasmalemma and tonoplast [9]. In its turn PA on the tonoplast is developed more slowly in comparison with plasmalemma. PA on the plasmalemma precedes the PA on the tonoplast. Conductivity of the membrane at the PA generation is increased, reaching its maximum in the peak of PA, then returns to the initial level. As plasmalemma so tonoplast behave, it is
14
Jaba Oniani, Vladimir Yurin, Ramaz Gakhokidze et al.
shown in figure 1. But for first of them it is characterized the considerable alternation of potential and conductivity.
Figure 1. Development of the wave of PA on the chariophytes cell: a) between vacuole and external media, b) on the tonoplast, c) on the plasmolemme.
Independent of outer conditions (environment composition, temperature, light, etc.) and also of physiologic conditions of the cells, the potential of action has a different amplitude and duration. Likely in the case of nerve fiber, PA can exceed potential zero value, i.e., there can be observed overshoot. Cells, independent of medium conditions, physiologic state, etc. can generate PA difference form and value [8,9]. At the end of the XVIII century, it was stated that at a weak electrical current the movement stops in the cytoplasm of seaweed Characea [10]. For the first time, the movement of the cytoplasm of plant cells, which later received the name cyclosis, was observed by Corti in 1774 [11]. At the accumulation of facts they came to the conclusion that cytoplasm movement occurs in the many intact cells, and the character of movement is quite various [12-16]. Reports stated the presence of intercellular movement cytoplasm along plasmodesm, in particular in the cells of parenchyma of different Allium sativum tissues. It is supposed that with the cytoplasm current there is carried away such low molecular substances as sugar amino acids and inorganic salts, etc. [12]. There exists a dependence between growth and movement direction of cytoplasm. Most strong growth of fungi floccus is marked when cytoplasm moves to the top direction. Probably moving cytoplasm carries the substances necessary for floccus growth. If the movement had been stopped, the growth practically was being discontinued [12]. It is supposed that the translocation of generative and vegetative nucleuses at the pollen tube of covered semen plants happens due to movement of cytoplasm. It is clear that in this case the cyclosis has a great importance for impregnation [17]. In the experiments series the movement intensification of cytoplasm serves as an index of particular condition of the cell or
Reception of Physiologically Active Substances by Plasmatic Membrane…
15
proceeds of cell transition in this condition. Researchers observed the activation of the cyclosis in the cell of some plants after implantation of parasite fungus [18]. As an example of interconnection of cytoplasm movement intensity and physiologic state in the natural condition can be served the locked cells of stomas. At the closed stoma in the locked cells of the leaf Vicia there had been observed intensive movement, and in the open ones the cytoplasm did not move or there was revealed only oscillatory movement in a separate section ]12]. In accordance with the works of some researchers, the cells and tissues of different plants have different physiological and physical and chemical characteristics depending on the age and closeness to apical and basal ends of the plant. It brings us to the thought about the presence of distinctions in speed of cyclosis of those cells. In reality, it is shown that the most high value of cytoplasm movement speed is characterized for upper growing cells (about 40 mkm/s) of seaweed Characea Nitella, the lowest is for inferior ones. Difference in the values between first and fourth cells was the rate of 10 mkm/s, the tendency of cyclosis speed decrease from upper to down was expressed quite distinctly. These results, apparently, can be interpreted as an effect of metabolism of different level in the cells of different age; the young cells have most high cyclosis velocity —as a reflection of high activity of metabolic processes [19]. Consequently, the velocity of cytoplasm movement is connected with physiologic conditions of the cells and metabolic processes flowing in them. An electrical field influencing itself on the physiologic processes in the cells, affects the different kinds of movement into the plant [20] and as it was mentioned, on the cyclosis velocity also. Current influence on the cytoplasm movement strongly depends on its intensity, durability, etc. At the effect of significant outer electrical current and at the appearing of PA, cyclosis stoppage occurred. At first this phenomena was revealed on the seaweed Characea Nitella cells [21]. Interconnection between PA and cytoplasm movement stoppage were stated for different kinds of seaweed Characea [22-25]. It was revealed that cytoplasm movement stopping takes place after 1–2 sec. of PA beginning or through 1 sec. after PA peak [25-27]. Complete restoration of cyclosis velocity occurs after 5–10 min [26]. This time depends on the kind of object and external conditions [24,25]. For example, in the autumn cells, the movement speed had decreased sharply at the cell excitation, was not stopped completely and then was being turned to the initial level [25]. It must be noted that appearance of PA is always accompanied by stopping or deceleration of the cyclosis but it is not necessary to have a sharp stopping of movement for the appearance of PA. Velocity of cytoplasm movement depends on contents of ATP and is inhibited by intercellular Ca2+ [28, 29]. At the injection of protein of akvorin in the cytoplasm of cells of Chara and Nitella, luminescence intensity of which is proportional to the concentration of free Ca2+ showed that the peak of intensity of luminescence was corresponding to the moment of cyclosis stopping and it had place at the maximum of the sluggish phase of PA. Concentration of free Ca2+ was being changed from 0.1–0.2 mcM to 6.7 mcM, in the peak of PA for the Chara cells, but for Nitella cells—from 0.44–1.1 to 43.0 mcM [28-30]. Increase of inter-cellular concentration of Ca2+ from 10-7 to 10–6 M at the absence of tonoplast notably slowed down the cyclosis in the cells of Nitella; discontinuance of the movement begins in at 10-3 M Ca2+. For the analogous fragments of Chara cells of critical concentration, the inhibited movement was 5·10-4 M. At the calcium removal had been observed the partial restoration of movement velocity [28,31,32].
16
Jaba Oniani, Vladimir Yurin, Ramaz Gakhokidze et al.
Due to the data mentioned above, we can suppose that the factor causing cytoplasm movement stopping at the actuation of membrane is Ca2+. This is confirmed that the fragments of the cells, without tonoplast at the introduction of EDTA complexon, connecting the Ca2+ was not observed at decrease of cytoplasm movement at the excitation of the cell [33]. It has been stated in the early works that in the cells of seaweed Characea the motive force is formed at the interaction between flowing endoplasm and immovable cortical layer and for the motion of cytoplasm it is necessary to have a definite organization of the cortical layer [34-35]. Afterwards were obtained a number of data touching on the presence of subcortical fibrils and endoplasmatic filaments. The last, as it was shown, are the branches of subcortical fibrils [36-39]. By such a method, it is possible to have a double action of calcium ions on the cyclosis. Such action at first is the reversible inhibition Ca2+ of sensitive mechanisms. Secondly is the inhibition through it acting on the filament [32]. Supposition about cytoplasm movement in the vegetable cells connecting with functioning of retractive proteins partially was confirmed after disclosure of actin in the higher plant Ammaryllis [40]. But the earlier works [41] indicated the Myozin similar character of extract obtained by cells of Nitella. More careful identification of protein extract from cells of Nitella has been conducted in the work [42]. At the significant ion force, the activity of identified ATphase protein complex by acting of Ca2+ or EDTA was being increased, but under influence of Mg2+- decreased. Purified preparation of the moizin from the cells of Nitella has had a higher molecular mass as compared to myosin, extracted from skeleton muscles. At last time myosin was identified also to the cells of other high plants, for example, Elodea [43]. Lucopersicon [44]. Extracted from all kind of plants myosin possessed the ability to activate actinstimulated ATphase and formed biopolymer aggregate [45-47]. One of the problem at the research of cytoplasm movement mechanism is an elucidation of intercellular myosin localization and construction its acting model. There was realized the separate processing of endoplasm and cortical layer by specific agents [48,49]. With this goal there were conducted the soft centrifugation intermodal cells of seaweed Characea, at which the cytoplasm was collected to the end of cell and the cortical layer, including the chloroplast and subcortical fibrils remained intact on the opposite (centripetal) end of the cell. Then one end of cell (cortical layer or endoplasm) were processed by corresponding agent and accomplished repetitive centrifugation, at which treated cytoplasm came into contact with non processed cortical layer. Processing of cellular cortical layers with N-ethylmaleinimide (EMI) inhibited with Factin activated myosin of ATphase did not influence the cyclosis. At processing of endoplasm there was not observed the movement. Differentiated holding of separate cell fragments during two minutes at the 47.5°C temperature was being led to acting analogous EMI. This can be explained that myosin is easy denaturizing at the temperature increase, while the actin is more thermostable. At application of cytohalasin B, inhibiting action of which on the cytoplasm movement in the vegetable cells is investigated well [50-53], an opposite effect was being noted as compared to above description. Cytoplasm movement was being stopped only after processing of cortical layer of cell by cytohalasin [54] at the acting of it on endoplasm the cyclosis was being continued. Adducted experimental data allowed authors to consider that cytohalasin, most likely inhibits the microfilaments of subcortical fibrils. Moreover they had
Reception of Physiologically Active Substances by Plasmatic Membrane…
17
stated the supposition that movable filaments of endoplasm do not play any role in the mechanism of protoplasm movement. But this contradicts the stated facts; there is shown by direct observation that the particles move in a quite similar way along endoplasmatic filaments and subcortical fibrils [13]. In connection with the abovementioned, most probable process providing generation of motive forces of cyclosis, is interaction, oriented to the movement direction and polarized actin filaments with citoplamatic oligomers of myosin [55]. Source of movement energy of cytoplasm is the conversion of chemical energy of hydrolyzes ATP by actinomyosin complex in conformation alteration of oligomers of myosin [55-58] which lead to the mechanical directed movement of cytoplasm. By analogy with acsoplasm there are considered that in the cells of seaweed Characea the places of filaments attachment to the excitable membrane are the protein molecules with high affinity to ions Ca2+In result of calcium ion entrance and their tying with active centers, there is weakened the connection between protomers of actin filaments, occur their unfastening from membrane, what leads to the cyclosis stopping. So there was pointed earlier that PA or potassium depolarization in the akson was being caused unfastening membrane connected filaments of cytoskeleton. Following removal of ions Ca 2+leads to fastening of actin filaments [59-60]. Such, there was appeared the circumstantial results, touching to mechanism of cyclosis beginnings in the vegetable cells and with DEP cells. For detailing of regularity of cyclosis interconnection and DEP, it was under-taken by us the attempt of their simultaneous measuring. As it is shown in fig.2, changing of DEP value, entail for them displacement of cytoplasm movement [61].
Figure 2. Kinetics of depolarisation (a) displacement of protoplasm movement (b) under the action of 3.0 mM (1) and 10 mM KCl (2).
For revealing of the qualitative character of this dependence, as an influencing factor on DEP, there was selected K+ ion. It is known that in the physiologic conditions, content this ion in the cytoplasm (10-150mM) of the vegetable cells are 2-3 orders above then in the environment. Therefore the little changes its concentration in the outer solutions (from 0,1 to
18
Jaba Oniani, Vladimir Yurin, Ramaz Gakhokidze et al.
10,0 mM) will not influence essentially on the concentration in the cytoplasm and the same it do not act directly on the structure, responsible for cyclosis process. Considering this displacement appearing in this conditions of DEP (ΔΨ) as the single factor, causing cyclosis speed changing (ΔV), there can be supposed as first approximation, that velocity induced ΔΨ alteration V will be proportional of it. But it must be taken into consideration the presence of opposite, adaptation tendency, proportional to the current displacement ΔV: dV/dt = k1ΔΨ (t) – k2ΔV(t)
(1)
Or after integration t ΔV(t)= k1 / k2 exp( k2 t) ∫ ΔΨ (τ)exp( k2 τ) dτ
(2)
0
Kinetics of Changing DEP and cytoplasm movement speed were approximated by equation of (2). Obtained equation describes well the experimental dependence (see fig. 2), what allows us to use it for estimation of potential dependence of cyclosis speed displacements, coming by acting of different factors [61,62]. In the frame of this model, in case, when experimental points on diagram (ΔV, ΔΨ) is placed on the straight line, passing on the origin of the coordinate system, the reaction is strongly potential dependent (Fig.3).
Fiigure 3. Connection between means of displacement ΔV and corresponding them displacement ΔΨ in case of potential dependence of VPM depending on relation (2).
Reception of Physiologically Active Substances by Plasmatic Membrane…
19
If the condition of analyzing set of points essentially declines from described dependence – this means that in considered case there has place the effects stipulated by interaction of effector with regulating cyclosis velocity by structures directly. Those, there is possible in the every separate case to determine how the observed effects are realized by overregulation of membrane potential, i.e. in sum it is realized by changing of membrane penetration, and in which - due to presence other acting way (for example, through influence on the different intercellular processes). For clarification of possible presence on the plasmatic membrane of vegetable cells structures, similar animal receptors and character of interaction with them effector, by us were conducted fundamental research of acetylcholine acting and number of biogenic amines on the speed of cytoplasm movement and DEP of cells of seaweed Characea [62,63]. Acetylcholine with concentration of 5·10-7 - 5·10-6M caused cyclosis velocity decrease and drop of DEP, which gradually was being turned to the initial level. More expressed action of acetylcholine was being provided at the concentration 5·10-5 M. Reaction of the cell was reversible i.e after removal it from solution was restored the initial movement velocity of cytoplasm. At cumulative acting of increased concentration of acetylcholine on the cell in the interval 5·10-7-5·10-5 M the concentration caused plasmolysis. Serotonin was being provided the opposite action of acetylcholine on the cyclosis speed. Process of cytoplasm movement at the serotonin action bore the oscillatory character. Concentration of serotonin in the medium till 10-5 M caused cyclosis speed raise while at the action of more high one, (10-4 M) the velocity decreased and the DEP meaning was being fallen. Adrenaline and noradrenaline was being begun its influence on the cyclosis and DEP of Nitella cells at the concentration 10-7 M and above it, at that the changes bore oscillatory character. Cell treatment by isodrin, which is stimulator of β adrenoreceptors in the animal organism at the concentration 10-6 M was being led to the increase of cytoplasm movement speed and DEP of the cell. Inderal (blockader of β adrenoreceptors) at concentration 10-6 M caused rapid decrease of cyclosis and DEP velocity. At the increase of direct effect (acting of testing agent on the intercellular system, responsible for keeping of cyclosis) on the movement of acetylcholine cytoplasm and biogenic amines arrange in the row: phenitron
