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Biomedical subjects

V Brabec

Publications and source records attributed to V Brabec.

At least 145 records · Page 8Linked to original sources

An electrochemical analysis of changes in properties of DNA caused by ionizing and ultraviolet radiations.

Electrooxidation and electroreduction of gamma- and u.v.-irradiated DNA were studied by means of differential pulse voltammetry at the graphite electrode and differential pulse polarography at the dropping mercury electrode. Two separated voltammetric oxidation peaks G and A were used for monitoring conformational changes in guanine . cytosoine (GC) and adenine . thyminde (AT) pairs respectively in irradiated double-stranded (ds) DNA. Pulse-polarographic reduction peak III was used for detection of denatured DNA in the irradiated samples of ds DNA. It was found that the heights of peaks G and A of ds DNA did not change with the radiation dose after relatively low doses of gamma- and u.v.-radiations (up to ca. 40 krads and 1 X 10(4) Jm-2, respectively, when no single-stranded (ss) DNA was detected in the irradiated DNA samples. After higher doses of radiation the occurrence of ss DNA or ss segments in the irradiated samples of ds DNA was accompanied by an increase of peaks G and A; however, peak A grew more rapidly with the increasing dose than peak G. It was concluded that the results obtained support the assumption, according to which regions of ds DNA rich in AT pairs are more susceptible to denaturation caused by gamma- and u.v.-radiations.

Cobalt Radioisotopes↗

Electrochemical behaviour of proteins at graphite electrodes. II. Electrooxidation of amino acids.

Electrochemical oxidation of L,alpha-amino acids at a paraffin-wax impregnated spectroscopic graphite electrode (WISGE) was studied by means of linear sweep, cyclic, phase-sensitive alternating current and differential pulse voltammetric techniques. It was found that out of the amino acids usually occurring in proteins only tyrosine, tryptophan, histidine, cystine, cysteine and methionine were oxidized at the WISGE. At relatively low concentrations of amino acids (up to ca. 2 x 10(-4) M) the electrode process in which the amino acids are oxidized at the WISGE has the characteristics of an irreversible reaction controlled by diffusion. Coulometric measurements showed that oxidation of tyrosine and tryptophan at the WISGE, i.e. of amino acids which are responsible for the oxidizability of proteins at graphite electrodes, is a two-electron process. At higher concentrations of tyrosine-and tryptophan (above ca. 2 x 10(-4) M) adsorption of the oxidation product of these amino adds was demonstrated.

Journal Article↗

Interaction of nucleic acids with electrically charged surfaces. VII. The effect of ionic strength of neutral medium on the conformation of dna adsorbed on the mercury electrode.

Triangular-wave direct current (d.c.) voltammetry at a hanging mercury drop electrode and phase-selective alternating current (a.c.) polarography at a dropping mercury electrode were used for the investigation of adsorption of double-helical (ds) DNA at mercury electrode surfaces from neutral solutions of 0.05-0.4 M HCOONH4. It was found for the potential region T (from -0.1 V up to ca. -1.0 V) that the height of voltammetric peaks of ds DNA is markedly influenced by the initial potential only at relatively low ionic strength (mu) (from 0.05 up to ca. 0.3). Also a decrease of differential capacity (measured by means of a.c. polarography) in the region T depended markedly on the electrode potential only at relatively low ionic strength. The following conclusions were made concerning the interaction of ds DNA with a mercury electrode charged to potentials of the region T in neutral medium of relatively low ionic strength mu < 0.3). (i) When ds DNA is adsorbed, a significantly higher number of DNA segments is anchored in the positively charged electrode surface than in the surface bearing a negative charge, (ii) In the region T, especially adsorbed labile regions of ds DNA are opened in the electrode surface, which are present in ds DNA already in the bulk of the solution, (iii) In the narrow region of potentials in the Vicinity of the zero charge potential a higher number of ds DNA segments can be opened, probably as a consequence of the strain which could act on the ds DNA molecule in the course of the segmental adsorption/desorption process.

Journal Article↗

Agglutinating phenomenon in the rat sera after administration of methylcellulose.

In the serum of Wistar rats following intraperitoneal application of methylcellulose (Kabi) the presence of the aggregation factor was confirmed. Using the more sensitive immunohaematological method-enzyme tests-we proved the agglutination phenomenon, which can be of antibody nature and cannot be induced under the test conditions in vitro by action of methylcellulose on blood cells and serum of normal rats. The described phenomenon will only occur if methylcellulose "passes" the organism of the experimental animal.

Animals↗

Interaction of nucleic acids with electrically charged surfaces. VI. A comparative study on the electrochemical behaviour of native and denatured DNAs at graphite electrodes.

Adsorption and electrochemical oxidation of deoxyribonucleic acid (DNA) at a pyrolytic graphite electrode (PGE) and a paraffin wax-impregnated spectroscopic graphite electrode (WISGE) were studied using differential pulse voltammetry. DNA is adsorbed at the surface of the graphite electrodes in a broad range of potentials including the potentials of electrochemical oxidation of DNA. Both native and denatured DNAs yield two single, well-defined and separated peaks, G and A, on the differential pulse voltammograms at the PGE and WISGE. The more negative peak, G, corresponds to electrochemical oxidation of adenine residues. Peaks G and A of native DNA occur at the same potentials as peaks G and A of denatured DNA. However, electrochemical oxidation of adenine and guanine residues at graphite electrodes is markedly suppressed in native DNA. The heights of the peaks G and A represent a sensitive indicator of the helix-coil transition of DNA. An analysis of the product of interaction of a sample of native DNA with a large pyrolytic graphite electrode in the presence of formaldehyde at approximately neutral pH did not prove changes in the secondary structure of native DNA due to its interaction with the graphite electrode. It is suggested that the decreased differential pulse-voltammetric activity of native DNA is connected with its decreased flexibility.

Animals↗

The effect of intravenous injection of ethyl palmitate emulsion on sequestration of thermally damaged erythrocytes in rats with experimentally induced hypersplenism, and in normal rats.

The authors attempted at experimental elimination of sequestration function of the spleen in Wistar rats using an i.v. injection of ethyl palmitate emulsion, both in "hypersplenic" animals being long-term applied i.p. methyl cellulose solution, and in control rats. In the rats clearance of 51Cr-labelled and thermally damaged erythrocytes from blood was examined and their sequestration in the spleen and liver followed. The ethyl palmitate injection resulted in both experimental groups in a significant decrease of the erythrocyte counts sequestrated in the spleen, and significant prolongation of the elimination half time for thermally damaged erythrocytes from the blood.

Animals↗

Temperature dependence of the interfacial behavior of uracil derivatives.

The effect of temperature on the interfacial behavior of uracil, thymine and 1,5-dimethyluracil has been used to characterize the thermodynamics of adsorption of these compounds at a mercury electrode-aqueous electrolyte interface. Between 4.5 degrees C and 40 degrees C all three compounds exhibit two adsorption regions. The first, a dilute layer where the molecules adsorb in a flat orientation on the electrode surface. The second, a compact layer where the molecules adsorb in a perpendicular orientation. The thermodynamic constants characterizing formation of these surface layers have been deduced.

Adsorption↗

Potential and structure controlled interfacial behavior of uracil derivatives.

The adsorption and related interfacial behavior of uracil, various methylated uracil derivatives, uridine, uridine-5'-monophosphate and uridine-3'5'-cyclic monophosphate has been studied by surface electrochemical measurements at a mercury electrode. All uracil derivatives exhibit an initial "dilute" adsorption region where the virtually flat uracil residue is absorbed flat on the electrode surface. In the case of uracil and its methylated derivatives the area occupied by one molecule is about 60-70 A2. Uracil, thymine and 1,5-dimethyluracil exhibit a second adsorption region where they rearrange on the surface and adopt a perpendicular orientation and occupy about 40 A2 per molecule. In this perpendicular orientation the uracils are bound to the electrode through the N(3)-H or perhaps N(1)-H functions in a manner similar to their Watson-Crick bonding in nucleic acids. When in the perpendicular orientation the adsorbed molecules undergo extensive stacking (association) interactions, again similar to those observed between adjacent bases in nucleic acids. The ability of a uracil derivative to undergo a surface reorientation is critically dependent on electrode potential, bulk-solution concentration and molecular structure.

Electrodes↗

A new simple method for determination of erythrocyte filtrability.

A new simple filtration technique designed for measuring red cell filtrability in the routine laboratory use was developed. The suspension of the whole blood in saline (1:20,000 dilution) was processed on the Sartorius filter membranes, pore size 8 micron. The percentage of passed erythrocytes indicating red cell filtrability was determined. The suitability and perspective applicability of this method for studying various hematological disorders is proposed.

Diagnosis, Differential↗

Lysis of red blood cells in some haemolytic anaemias--the lytic effect of agkistrodon piscivorus venom in vitro.

The lysis of red blood cells induced by the Agkistrodon piscivorus venom (APV) in vitro in the presence and/or in the absence of plasma was examined in autoimmune haemolytic anaemia (AIHA), hereditary spherocytosis (HS) and paroxysmal nocturnal haemoglobinuria (PNH). Virtually haemolysis did not differ from that of normals in AIHA, it was slightly increased in HS and significantly higher in PNH, however, only in the presence of autologous or normal homologous plasma. The mechanism of PNH blood cells lysis which is probably related to the activation of the third complement component is discussed.

Anemia, Hemolytic↗

Interaction of nucleic acids with electrically charged surfaces. II. Conformational changes in double-helical polynucleotides.

The influence of adsorption of double-stranded (ds) DNA, ds RNA and homopolymeric pairs at a mercury electrode on conformation of these polynucleotides was studied. Changes in the polarographic reducibility of polynucleotides, which were followed by means of normal pulse polarography and linear sweep peak voltammetry at the dropping mercury electrode were exploited to indicate conformational changes. It was found that, as a consequence of adsorption of ds polynuclotides on the negatively charged electrode conformational changes similar to denaturation take place in a narrow potential region around -1.2 V (the region U). After sufficiently long time of the contact with the electrode (under our conditions about 10 s) these changes reach limiting values, which can approach total denaturation. Upon adsorption of ds polynucleotides on the electrode charged to more positive potentials than the region U either (1) no conformational changes occur or (2) only a small part of the polynucleotide (probably labile regions of the ds molecule) is very quickly denatured - the remainder of the molecule preserves its ds structure. Conformational changes of adsorbed ds polynucleotides are influenced by factors which change the stability of ds polynucleotides in solution. It is supposed that denaturation of ds polynucleotides in the region U might result from the strains connected with the repulsion of certain segments of the molecule anchored on the electrode from the negatively charged surface.

Animals↗