Biomedical subjects

F Lisdat

Publications and source records attributed to F Lisdat.

11 recordsLinked to original sources

PubMed2004

Comparing an in vitro electrochemical measurement of superoxide scavenging activity with an in vivo assessment of antioxidant potential in Chinese tonifying herbs.

The in vitro superoxide scavenging activity (as determined by electrochemical measurement) and the in vivo antioxidant potential (as determined by a mouse model of carbon tetrachloride (CCl(4)) hepatotoxicity) of methanolic extracts prepared from 10 Chinese tonifying herbs were compared. Electrochemical measurement using a cytochrome c (Cyt. c) sensor showed that all of the tested herbal extracts exhibited a medium superoxide scavenging activity of different potency, as indicated by their IC(50) values. The in vivo measurement demonstrated that 80% of the herbal extracts displayed in vivo antioxidant potential, as assessed by the percentage of protection of the activity of plasma alanine aminotransferases and the hepatic glutathione regeneration capacity under CCl(4)-intoxicated condition. Although the in vitro antioxidant activity did not correlate quantitatively with the in vivo antioxidant potential, for 8 out of 10 samples a similar tendency was found. The rapid amperometric assessment of antioxidant potential by Cyt. c sensor may offer a convenient and direct method for screening as well as the quality control of herbal products.

Animals↗

PubMed2003

Epigallocatechin gallate can significantly decrease free oxygen radicals in the reperfusion injury in vivo.

OBJECTIVE: There is experimental evidence that oxygen-derived free radicals (Superoxide (O(2)(-))) play a key role in tissue damage in ischemia-reperfusion injury. Among various antioxidants tested in vitro, natural polyphenols like Epigallocatechine gallate (EGCG) show a 164-fold higher scavenging activity for O(2)(-) than ascorbic acid We therefore conducted an animal study in order to investigate the impact of EGCG on O(2)(-) production during reperfusion after defined periods of ischemia in the muscle tissue of the rat, using a recently developed cytochrome c-based biosensor for on-line in vivo monitoring of O(2)(-). MATERIALS AND METHODS: Femoral artery and vein were dissected below the inguinal ligament in male Wistar rats. The cytochrome c-based biosensor was placed in the gastrocnemius muscle. Ischemia was induced by clamping the femoral vessels. Ischemia times were either 60 (n = 14) or 120 (n = 14) minutes. Six animals in each group received 4 mg/kg body weight EGCG intravenously at the time of reperfusion, another six animals in each group served as controls (no treatment). Additionally, two animals in each group received the same volume of saline instead of EGCG. The current response of the biosensor corresponding to the O(2)(-) concentrations in vivo was recorded on a PC. The gastrocnemius muscles were harvested for histological evaluation. RESULTS: The average maximum O(2)(-) concentration after 60 minutes of ischemia was 188, 18 nmol/L (23 pA) compared to 90 nmol/L (11 pA) (P <.01) with EGCG application. The mean O(2)(-) value after 120 minutes was 220 nmol/L (27 pA) versus 135 nmol/L (16.5 pA) (P <.01) with EGCG, respectively. Histological analysis showed advanced muscle cell injury and neutrophil infiltration in the group without EGCG. No O(2)(-) reduction could be verified administering saline instead of EGCG. CONCLUSION: For the first time the scavenging activity of an antioxidant was verified in vivo on-line. EGCG significantly diminished O(2)(-) tissue concentrations after 60 or 120 minutes of ischemia by an average of nearly 50%, suggesting its therapeutic potential.

Animals↗

PubMed2003

Electrochemistry of immobilized CuZnSOD and FeSOD and their interaction with superoxide radicals.

Copper, zinc superoxide dismutase (CuZnSOD) from bovine erythrocytes and iron superoxide dismutase from Escherichia coli (FeSOD) were immobilized on 3-mercaptopropionic acid (MPA)-modified gold electrodes, respectively. The characterization of the SOD electrodes showed a quasi-reversible, electrochemical redox behavior with a formal potential of 47+/-4 mV and -154+/-5 mV (vs. Ag/AgCl, 1 M KCl) for surface adsorbed CuZnSOD and FeSOD, respectively. The heterogeneous electron transfer rate constants were determined to be about 65 and 35/s, respectively. Covalent fixation of both SODs was also feasible with only slight changes in the formal potential. The interaction of superoxide radicals (O(2)(-)) with the SOD electrode was investigated. No catalytic current could be observed. However, due to the fast cyclic redox reaction of SOD with superoxide, the communication of the protein with the electrode was strongly influenced. The amperometric detection of superoxide radicals is discussed.

Adsorption↗

PubMed2002

Amperometric biosensor based on a functionalized gold electrode for the detection of antioxidants.

A method for the electrochemical detection of antioxidants has been developed, which is based on a radical measurement with a cytochrome c modified electrode. A controlled enzymatic production system for superoxide radicals based on xanthine oxidase was used. The addition of antioxidants facilitated the decomposition of the radical in addition to the spontaneous dismutation. The steady-state of superoxide generation and decomposition was thus shifted to a new situation due to the higher decomposition rate after antioxidant addition. This resulted in a decreased current level at the electrode. Antioxidant activity could be quantified from the response of the sensor electrode by the percentage of the signal decrease. The 50% inhibition value (IC(50)) for different antioxidants was calculated and the antioxidant activity of numerous substances was compared. Thus, a hierarchy of superoxide radical scavenging abilities of flavonoids was established: flavanols>flavonols>flavones>flavonones>isoflavonones.

Antioxidants↗

PubMed2002

Copper proteins immobilised on gold electrodes for (bio)analytical studies.

Copper electrochemistry at modified gold electrodes was investigated with two different states of the metal ion: first bound in azurin from Pseudomonas aeruginosa and second introduced via metal ion uptake in metallothionein (MT) from rabbit liver. Azurin was immobilised on a mercaptosuccinic acid (MSA) layer self-assembled on gold. The redox behaviour in the adsorbed as well as in the covalently immobilised state was found to be quasi-reversible with a formal potential of +198 mV versus Ag/AgCl. The pH variation suggests an optimal pH range for efficient electrode communication in the neutral range. MT was fixed at electrochemically cleaned gold using the accessible cysteins of the protein. Copper was found to bind to the MT-modified gold electrode. The electrochemical behaviour of the bound copper was characterised in copper-free solution with a formal potential of +245 mV versus Ag/AgCl. Stability and potential use is discussed.

Animals↗

PubMed2001

An optical method for the detection of oxidative stress using protein-RNA interaction.

The cytosolic 4Fe-4S protein aconitase can be converted under the influence of reactive oxygen species into an iron-regulatory protein (IRP1). Therefore, the IRP1 level is considered as an indirect marker of oxidative stress. An experimental approach is presented here to detect the concentration of this marker protein by surface plasmon resonance. The optical method exploits the natural binding affinity of IRP1 to an iron-responsive element (IRE) which was in vitro transcribed with a linker sequence and subsequently immobilized on a BIACORE sensor chip. The detection was found to be reproducible and sensitive in the range 20-200 nM IRP. Conditions of the binding process, such as pH and thiol concentration, were characterized. Feasibility of the method to detect and quantify IRP1 in physiological media was demonstrated.

Aconitate Hydratase↗

PubMed2001

Research and development in biosensors.

Progress in biosensors has mainly been made by the improvement of the biological components and the implementation of microsystem technologies. Enzymes are still the most appropriate recognition elements because they combine high chemical specificity and inherent biocatalytic signal amplification. A breakthrough has been achieved in the application of membrane-integrated receptor systems for analyte recognition and signal transduction in biosensors. Sensor integration of RNA aptamers has been initiated, and the performance of fully synthetic molecularly imprinted polymers has been improved.

Antibodies↗

PubMed2000

Electrochemical investigation of cellobiose oxidation by cellobiose dehydrogenase in the presence of cytochrome c as mediator.

An important aspect of the cytochrome c electrochemistry is the possibility of coupling the 'heterogeneous reactions' with other redox enzymes. Cellobiose dehydrogenase, a 89170 Da glycoprotein that contains both FAD and a b-type haem as prosthetic groups, donates electrons to a number of acceptors, including cytochrome c. While haem b is surrounded mainly by acidic amino acids, cytochrome c displays positive charged lysine groups around the haem site. Thus a fast reaction between both proteins is explicable. In the presence of cellobiose, a catalytic current was observed, owing to the interaction of cellobiose dehydrogenase with electrostatically adsorbed cytochrome c. Adsorption of cytochrome c provides a technological model surface for vectorial electron transfer.

Carbohydrate Dehydrogenases↗

PubMed1997

Catecholamine detection using enzymatic amplification.

Different amplification sensors based on the substrate recycling principle were investigated with respect to their applicability to catecholamine detection. In the bioelectrocatalytic approach, glassy carbon electrodes were modified by laccase or a PQQ-dependent glucose dehydrogenase. Substrate recycling occurs and the detection limit is in the lower nanomolar concentration range (e.g. 10 nM dopamine and 1 nM noradrenaline for the laccase- and glucose dehydrogenase-modified electrodes, respectively). Combinations of glucose dehydrogenase with laccase or tyrosinase were investigated as bienzymatic probes. Among the systems we studied, the laccase/glucose dehydrogenase sensor is the most sensitive (detection limit: 0.5 nM adrenaline). The selectivities of the different sensor systems are discussed. Application of the laccase/glucose dehydrogenase electrode in different media (i.e. brain homogenate, heart effluate) was successfully shown. For samples with high concentrations of interfering substances (uric and ascorbic acid), the interferences can be effectively removed using enzymatic methods.

Biosensing Techniques↗

PubMed1997

Enzymatic substrate recycling electrodes.

A weak chemical signal might result in a large response when biochemically amplified. Enzymatic recycling of the analyte is one of the biochemical ways of providing an effective increase in biosensor sensitivity by several orders of magnitude. The enhancement of sensitivity is provided by consecutive consumption and generation of the analyte on the sensor surface. The principle of enzymatic substrate regeneration using bioelectrocatalysis and coupled enzymes is shortly reviewed and illustrated with some recent developments of biosensors for catecholamines, and its potential for electrochemical immunoassays is outlined.

Biosensing Techniques↗

PubMed1997

Ultrasensitive bienzyme sensor for adrenaline.

A biosensor consisting of an analyte-recycling two-enzyme system using laccase (Coriolus hirsutus) and PQQ-dependent glucose dehydrogenase in combination with the electrochemical detection of oxygen depletion at a platinum electrode was used for adrenaline determination in the nano- and subnanomolar concentration range. Measurements were performed in a flow cell providing excellent baseline stability and fast recovery of the sensor. Improved design of the polymer matrix resulted in a lower detection limit of 200 pmol/l for adrenaline. The sensor has successfully been applied to the analysis of adrenaline in effluate of isolated rabbit hearts.

Animals↗