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The reactions of the 2'-deoxyadenosine electron adduct in aqueous solution. The effects of the radiosensitizer p-nitroacetophenone. A pulse spectroscopic and pulse conductometric study.

The reactions of the one electron adduct of 2'-deoxyadenosine dA(-), in aqueous solutions have been studied using pulse radiolysis techniques with optical absorption and conductivity detection. The dA(-) radical anion itself shows a weak and featureless optical absorption at greater than 300 nm. It reacts rapidly with H20 (k equals 6 x 10(4)M(-1)s(-1); t1/2 equals 210 ns) to yield at least three different protonated structures (dAH., dA'H., dA"H.). In neutral solutions the most important of these (dAH., lambda(max) equals 315 nm) decays by a first order process (t1/2 approximately equal to 9 microseconds). In basic solutions dAH. undergoes on OH- catalysed rearrangement into another neutral radical (dA"'H., lambda(max) equals 355 nm; k(dAH. plus OH-) equals 1.7 x 10(8)M(-1)s(-1)). p-Nitroacetophenone (PNAP) reacts rapidly with the protonated electron adducts of 2'-deoxyadenosine (k equals 5 x 10(9)M(-1)s(-1)). An electron transfer occurs with dAH to yield PNAP and a reoxidized form of 2'-deoxyadenosine. As indicated by its pK value of 8.8 the latter is not, however, simply a repaired 2'-deoxyadenosine molecule, but is suggested to include the elements of water. Species dA"'H.(and dA'H.) react with PNAP in a process which is clearly not an electron transfer but likely an addition reaction.

Acetophenones↗

Selective interaction of tirapazamine with DNA bases and DNA. A comparison of cyclic voltammetry and electrolysis techniques.

An electrochemical model has been used to study the reductive activation of the hypoxic cell cytotoxin tirapazamine (TPZ, 3-amino-1,2,4-benzotriazine-1,4-dioxide). Cyclic voltammetry and controlled potential electrolysis have been used to generate and study the 1-electron reduction product, the assumed biologically active species. Cyclic voltammetry of tirapazamine in dimethylformamide shows a quasi-reversible 1-electron reduction with the product showing a tendency to participate in a following chemical reaction. Controlled potential electrolysis to generate the 1-electron reduction product was unsuccessful due to the formation of a new redox-active species at less negative reduction potentials. However, the cyclic voltammetry of tirapazamine in the presence of E. coli DNA shows a decrease in the lifetime of the radical anion, signifying direct interaction with the DNA. The radical lifetime also decreased in the presence of adenine, thymine and guanine, but increased upon addition of cytosine and ribose. The study shows that cyclic voltammetry is an extremely useful tool for investigating the interaction between bio-reductive drugs and biological target molecules.

Adenine↗

An amperometric enzyme electrode for creatine determination prepared by the immobilization of creatinase and sarcosine oxidase in poly(vinylferrocenium).

A new enzyme electrode for the determination of creatine was developed by immobilizing creatinase (CI) and sarcosine oxidase (SO). The enzymes were co-immobilized in a poly(vinylferrocenium) matrix onto the surface of a platinum working electrode. Crosslinking with glutaraldehyte (GA) and bovine serum albumin (BSA) was selected as the best immobilization method for the enzymatic system. Determination of creatine was performed by the oxidation of enzymatically generated H2O2 at + 0.7 V vs. Ag/AgCl. The linear working range of the electrode was 2.0 x 10(-5) - 3.2 x 10(-4) M and the response time was about 50 s. The effects of pH, temperature, enzyme ratio and buffer concentration were investigated and optimum parameters were found to be 7.5, 37 degrees C, 2.5:1 (CI:SO) and 0.05 M, respectively. The stability and reproducibility of the enzyme electrode have been also studied.

Biosensing Techniques↗

Electrochemical study on behavior of EuMo2 complex and its interaction with DNA.

The electrochemical behavior of complex EuMo2 (Mo = Morin, 2',3,4'5,7-pentahydroxyflavone) and its interactions with calf thymus DNA were studied using cyclic voltammetry (CV) and double potential step chronocoulometry (DPSCC) at glass carbon electrode (GCE) and DNA modified GCE, respectively. Information such as diffusion coefficient (D), rate constant (ks) of EuMo2 and intrinsic binding constant (K), binding numbers (n) of bound species per DNA (bp) were obtained. EuMo2 can bind to DNA, and the binding mode is intercalation. By nonlinear fitting with Langmuir equation, a K of 1.02 x 10(6) M-1 and an n of 1 were obtained.

Animals↗

Right ventricular conductance to establish closed-loop pacing.

Innovations in pacing technology, which include the addition of rate-responsive features to programmable pacemakers, can improve the quality of life of patients suffering from sick sinus syndrome. Among the strategies providing rate-adaptive cardiac pacing, the most attractive is the physiological restoration of closed-loop chronotropic control. This paper describes how autonomic nervous system (ANS) control information is extracted from dynamic measures of myocardial contractile performance obtained from unipolar conductance measurements using the stimulation electrode in the right ventricular cavity. The pacemaker uses the ANS information to modulate pacing rate and restore normal physiological control of heart rate. A new algorithm, regional effective slope quantity (RQ), for isolating the ANS signal was developed. The resulting signal, ventricular inotropic parameter (VIP), is a normalized parameter proportional to the strength of the ANS inotropic signals to the myocardium. The efficacy of the ANS control concept was evaluated in multi-centre studies. Patients with AV block and VIP-controlled pulse generators performed defined exercise protocols. The ANS-controlled pacing rate and the spontaneous sinus rate were closely correlated. Blood pressure and subjective patient reports further indicated that good control of the cardiovascular circulation was achieved.

Autonomic Nervous System↗

Bio-electric conductivity potentials in experimental skin grafts.

Prior investigations have established that changes in bio-electric potentials accompany the processes of wound creation and healing. In order to investigate these changes in an experimental full-thickness skin graft model, grafts were harvested from the dorsa of eight albino rabbits. Changes in potential were recorded over a period of 32 days, using silver-silver chloride electrodes and a recording polygraph. The potential measured across the skin graft became increasingly more electropositive until, between days 1 and 2, the potential abruptly reversed polarity. This negative potential lasted until day 4, when the conductivity again became positive, with a slow return to baseline measurements by day 32. Although these observations may indicate a bio-electric counterpart to the cellular events of wound healing, more study is needed.

Animals↗

Niacin assay by monitoring changes in electrical conductance caused by microbial growth.

A computerized Malthus 128 H Growth Analyser was used to determine the changes in conductance produced by growth of a niacin-dependent strain of Lactobacillus plantarum ATCC 8014 in different concentrations of niacin in the growth medium. Changes in conductance varied linearly with the niacin concentration in the range 5-50 ng/ml. The time required to complete a test is inversely related to the inoculum size, and results can be obtained after 6 h.

Conductometry↗

Selection of pH buffers for use in conductimetric microbiological assays.

Many metabolic activities of micro-organisms lead to changes in the pH value of cultures and consequently pH buffer compounds are potentially a major source of conductivity changes in cultures. To maximize changes in conductivity associated with microbial growth the pH buffer-associated changes should occur in a direction that reinforces those due to other metabolic activities. In agreement with this, studies with Escherichia coli showed that fermentation of glucose and aerobic growth on L-alanine yielded greater changes in the conductivity of media containing Tris(hydroxymethyl)aminomethane or L-histidine buffers than in a medium containing phosphate buffer, whereas aerobic growth on glucose or succinate yielded greater changes with phosphate buffer than with Tris(hydroxymethyl)aminomethane or L-histidine buffers. Criteria for the selection of appropriate pH buffer compounds are presented.

Aerobiosis↗

An investigation of indirect conductimetry for detection of some food-borne bacteria.

Indirect conductimetry using a rapid automated bacterial impedance technique was investigated. Strains of Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Aeromonas hydrophila and Salmonella spp. grown in Whitley Impedance broth all elicited indirect conductimetric changes. These indirect conductance responses were improved by the addition of 2 g/l glucose to the medium and resulted in maximum changes of 2340-4300 microS with associated maximum rates of change of 520-1210 microS/h. Furthermore, the indirect conductimetric assay detected growth of staphylococci, listeria and salmonella in media containing high concentrations of salts used as selective agents in culture media for the isolation of these organisms.

Aeromonas↗

The effect of the olive phenolic compound, oleuropein, on growth and enterotoxin B production by Staphylococcus aureus.

The presence of low concentrations (0.1% w/v) of oleuropein, a phenolic compound extracted from olives, delayed the growth of Staphylococcus aureus in NZ amine A and brain heart infusion media modified by the addition of growth factors and glucose (NZA+ and BHI+), as indicated by changes in conductance, whilst higher concentrations (0.4-0.6% w/v) inhibited growth completely. Intermediate concentrations of oleuropein (0.2%) prevented growth in BHI+ but allowed growth to occur in NZA+ despite an extended lag phase (30 h). Concentrations of oleuropein > 0.2% inhibited growth and production of enterotoxin B in both types of media. Lower levels (0.1%) did not affect the final viable count and production of toxin in BHI+ but decreased the number of viable organisms and reduced the toxin production in NZA+ by eightfold. An increase in the concentration of oleuropein resulted in a decrease in the amount of glucose assimilated and consequently the amount of lactate produced. In addition, oleuropein prevented the secretion of a number of exoproteins. Addition of oleuropein during the exponential phase appeared to have no effect on the growth of Staph. aureus in NZA+.

Colony Count, Microbial↗

Quantitative evaluation of antifungal activity of metallic oxide powders (MgO, CaO and ZnO) by an indirect conductimetric assay.

AIMS: To evaluate antifungal activities of MgO, CaO and ZnO powders quantitatively by indirect conductimetric assay. METHODS AND RESULTS: Candida albicans NBRC1060, Saccharomyces cerevisiae NBRC1950, Aspergillus niger NBRC4067 and Rhizopus stolonifer NBRC4781 were used as test micro-organisms. The indirect conductimetric assay, in which the change in electrical conductivity of an alkaline solution (NaOH) is produced by absorption of CO2 from microbial metabolism, could offer a simple and rapid evaluation of the antifungal activity within 24-48 h. The conductivity curves obtained for MgO, CaO and ZnO were analysed using the growth inhibition kinetic model proposed by Takahashi for calorimetric evaluation, and the kinetic parameters and minimum inhibitory concentration ([I]100) could be determined. MgO and CaO powders exhibited the antimicrobial activities against all fungi used in this study and showed little differences between types of fungi. However, although ZnO powder inhibited fungal growth, the values of [I]100 were over 100 mg ml-1. CONCLUSIONS: Although a common method for evaluating antifungal activity requires over 5-7 days, the indirect assay could provide a rapid and quantitative evaluation of antifungal activity within approx. 2 days, and MgO and CaO were found to have antifungal activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The indirect assay can be applicable for simple and rapid evaluation of the antimicrobial activity of insoluble or slightly soluble materials with high turbidity such as antibacterial ceramic powders. Moreover, these materials can be useful for controlling fungi in food processing and the environment.

Antifungal Agents↗

A new detector for fully automatic peptide synthesis.

A new method of monitoring the rate of reactions in solid phase peptide synthesis is described. A conductivity detector in the reaction cell enables the deprotection, washing, and subsequent coupling stages to be examined in detail. The half lives of the reactions can be calculated and hence the optimum reaction times predicted. The aggregation of peptide chains and subsequent collapse of the resin is observed. Difficult sequences are sensed and appropriate action taken completely automatically.

Amino Acid Sequence↗

Peptide aggregates: a novel model system to study self-assembly of peptides.

Ordered aggregates of Val-Leu-Pro-Phe, tetrapeptide 1, have been found in aqueous solutions. Evidence for the formation of aggregates for the above peptide was obtained by conductometric, pH metric, UV and fluorescence spectroscopic techniques. Values of critical micelle concentration (CMC) for the above peptide obtained by these methods are in good agreement with each other. The formation of organized aggregates of the peptide is favoured upon increasing the temperature (viz. the process of aggregation is endothermic). The aggregation number has been determined at different temperatures. Values of delta G0m, delta H0m, delta S0m and delta C0p have also been estimated. Binding studies with the 8-anilinonaphthyl sulfonic acid (ANS) and pyrene indicate that the interior of the aggregate is nonpolar. There are two processes with regard to the change of thermodynamical parameters like delta G0m, delta H0m, delta S0m, delta C0p and aggregation number (N). In the first process (from 5 degrees C to 40 degrees C) the driving force for aggregation seems to be the positive entropy because of water release due to intermolecular association of ionic moieties. The second process (from 40 degrees C and above) is due to intramolecular ionic interaction. The chemical shifts of the amide protons of the peptide have been presented in the light of inter- and intramolecular hydrogen-bond formation, and forces implicated in aggregation for both the first and second processes.

Amides↗

High precision mixing of anesthetic gases based on a new principle.

A machine has been constructed for mixing O2 and N2O. It consists of: (a) a proportional pressure and thereby flow regulator at the inlets for O2 and N2O; (b) a digital gas mixer which determines the gas mixture; and (c) a rotameter to measure the outlet flow. The contents of mixtures obtained from the machine were measured with a quadropole mass spectrometer (at 2 and 5 1 min--1 with downstream pressures of 500 and 3000 Pa). The mean numeric difference between desired and registered vol% O2 varied between 0.3 and 0.5 vol% at the four conditions tested. The maximal deviation was 1.2 vol%. Five conventional machines in daily use at the hospital were also tested. The mean numeric difference for these machines varied between 1.1 and 2.7 vol% O2. The maximum deviation was 7.4 vol%.

Anesthesia, Inhalation↗