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Biomedical subjects

W Göpel

Publications and source records attributed to W Göpel.

At least 19 recordsLinked to original sources

"Electronic nose" detects major histocompatibility complex-dependent prerenal and postrenal odor components.

Mice prefer to mate with individuals expressing different MHC genes from their own. Volatile components presenting MHC-dependent odor types are present in urine and can be detected by mice, as shown by extensive behavioral studies. Similar odor types are suspected to influence human behavior as well. Although a recent report indicates that MHC expression influences the ratio of volatile compounds such as phenylacetic acid, so far no other means than studying the behavior of mice or rats has been available to assess odor types. Here, we report the ability of a gas sensor array (referred to as "electronic nose") to detect MHC-dependent odor types. The electronic nose consists of an array of chemophysical detectors, in our case quartz crystal microbalances and semiconducting metal-oxide sensors that change frequency or conductivity upon binding of very small numbers of individual molecules present in the gas phase of odorous fluids. The pattern of changes is characteristic for a particular smell. Our electronic nose distinguishes the urine odor types of MHC congenic mouse strains, MHC class I mutant mice, and HLA-A2 transgenic mice. In addition, MHC-dependent odor types can be detected in serum. The device also clearly differentiates between individual odor types of human sera from HLA homozygous individuals; however, HLA expression seems to have only a secondary influence. Thus, odor-type research can now be carried out with an objective and fast through-put system independent of behavioral studies.

Animals↗

Rapid detection of expansions by PCR and non-radioactive hybridization: application for prenatal diagnosis of myotonic dystrophy.

The mutation specific for myotonic dystrophy (DM) is an unstable expanded CTG repeat located in the 3'-untranslated region of the myotonin protein kinase gene. Expansion of the CTG repeat shows a positive correlation with the severity of the disease and increases in successive generations of DM patients. Children with the congenital form of DM show the most severe phenotype and have large expansions, usually > 1000 repeats. For pregnant women with DM, prenatal diagnosis of DM may be offered. To reduce the time between chorionic villus sampling or amniocentesis and final results of DNA analysis in these cases, a fast and efficient method has been developed. This method combines direct PCR analyses for normal alleles with a nested PCR system followed by non-radioactive hybridization with a single-stranded probe.

Adult↗

Maturational changes of lymphocyte surface antigens in human blood: comparison between fetuses, neonates and adults.

The objective of this study was to investigate the maturational changes of lymphocyte surface antigens during ontogeny from fetuses to adults using the proven whole blood lysis technique. Two-color flow cytometric analysis of lymphocyte surface markers was performed on 20 fetal blood samples obtained by cordocentesis, 70 cord blood and 30 adult blood samples. The leukocyte count and all T-cell subsets were highest in neonates compared to fetuses and declined into adulthood. In contrast, the percentage of CD2+ T cells (fetal blood 57% versus adult blood 82%; p </= 0.001), CD3+ T cells (fetal blood 52% versus adult blood 77%; p </= 0.001), CD4+ T cells (fetal blood 39% versus adult blood 50%; p </= 0.05) and CD8+ T cells (fetal blood 15% versus adult blood 24%; p </= 0.001) showed a significant increase from fetuses to adults. The absolute and relative amounts of B cells were highest in fetuses (fetal blood 547 x 10(6)/l, 18%) followed by a steady decline to adulthood (adult blood 243 x 10(6)/l, 13%; p </= 0.001). The discordance between the absolute and relative size of lymphocyte subpopulations emphasizes the consideration of both variables in the assessment of lymphocyte maturation. The presented data may contribute to a better knowledge of the maturation of the immune system.

Adult↗

Performance of GC-MS and MOSES II, a hybrid modular sensor system, for the quantitative detection of the evaporation of the insect repellent N,N-diethyl-m-toluamide from two different matrices.

The purpose of this work was to determine the evaporated amount of N,N-diethyl-m-toluamide (DEET), a commonly used insect repellent, from two different matrices medium chain triglycerides (MCT) and Polyethylene glycol 400 (PEG), with GC with mass sensitive detection (GC-MS) and a hybrid modular sensor system (MOSES II). Thus, the binary mixtures are equilibrated at 80 degrees C for 45 min and then an aliquot of this headspace is analyzed by GC-MS and a sensory system consisting of an array of eight metal oxide sensors and eight quartz crystal microbalances. Both analytical methods allow a sensitive detection of even small amounts of DEET in the headspace from binary mixtures with the MCT or PEG, respectively. The two additives are found to be very different concerning their behavior to the vapor phase; i.e., the lipophilic (MCT) delivers an almost constant headspace of DEET, while the hydrophilic matrix PEG releases smaller amounts of DEET, which continuously decrease when a multiple headspace extraction is performed. Furthermore, the results reveal that both analytical methods lead to comparable results with the MCT/DEET mixtures whereas differences were seen with mixtures containing PEG. This can be attributed to the interaction of volatile portions of PEG with the sensors of the MOSES II.

Autoanalysis↗

Effective use of molecular recognition in gas sensing: results from acoustic wave and in situ FT-IR measurements.

To probe directly the analyte/film interactions that characterize molecular recognition in gas sensors, we recorded changes to the in situ surface vibrational spectra of specifically functionalized surface acoustic wave (SAW) devices concurrently with analyte exposure and SAW measurement of the extent of sorption. Fourier transform infrared external-reflectance spectra (FT-IR-ERS) were collected from operating 97-MHz SAW delay lines during exposure to a range of analytes as they interacted with thin-film coatings previously shown to be selective: cyclodextrins for chiral recognition, nickel camphorates for Lewis bases such as pyridine or organophosphonates, and phthalocyanines for aromatic compounds. In most cases where specific chemical interactions--metal coordination, "cage" compound inclusion, or pi-stacking--were expected, analyte dosing caused distinctive changes in the IR spectra, together with anomalously large SAW sensor responses. In contrast, control experiments involving the physisorption of the same analytes by conventional organic polymers did not cause similar changes in the IR spectra, and the SAW responses were smaller. For a given conventional polymer, the partition coefficients (or SAW sensor signals) roughly followed the analyte fraction of saturation vapor pressure. These SAW/FT-IR results support earlier conclusions derived from thickness-shear mode resonator data.

Acoustics↗

Saline lavage with substitution of bovine surfactant in term neonates with meconium aspiration syndrome (MAS) transferred for extracorporeal membrane oxygenation (ECMO): a pilot study.

BACKGROUND: Meconium aspiration syndrome (MAS) is still a condition associated with a high mortality, and many patients require extracorporeal membrane oxygenation (ECMO) as rescue therapy. Beneficial effects of surfactant and perflubron lavage have been reported. However, pure surfactant supplementation has not been proven to be beneficial in the most severe forms of MAS. This study was performed to demonstrate an improvement in oxygenation in neonates transferred for ECMO and fulfilling ECMO criteria with a saline lavage and surfactant resupplementation. METHODS: Twelve newborns with MAS [gestational age 36-40 weeks, mean birth weight 3200 g, age 4-16 h, oxygenation index (OI) > 40] transferred for ECMO therapy were treated with saline lavage (5-10 cm3/kg body weight, as long as green colored retrieval was observed) and resupplementation with bovine surfactant (Alveofact, Boehringer, Ingelheim, Germany). The OI at admission and 3 h after this procedure was compared using the t-test for paired samples. ECMO was available as rescue therapy at all times. RESULTS: The OI decreased from 49.4 (SD +/- 13.3) to 27.4 (SD +/- 7.3), P < 0.01. The decrease was sustained in nine patients, three patients required ECMO and all patients survived. CONCLUSIONS: As MAS is a condition with parenchymal damage, pulmonary hypertension and obstructive airway disease, no simple causative therapy is possible. Surfactant application after removal of meconium by extensive lavage is feasible as long as 16 h after birth even in infants considered for ECMO therapy; it might reduce the necessity of ECMO.

Journal Article↗

Oxygenation and lung morphology in a rabbit pediatric ARDS- model under high peak pressure ventilation plus nitric oxide and surfactant compared with veno-venous ECMO.

UNLABELLED: The aim of the study is to investigate which of two treatment options of saline lavage induced ARDS in rabbits is better in terms of oxygenation and prevention of barotrauma: combined high peak pressure ventilation with surfactant administration and inhaled nitric oxide or veno-venous ECMO combined with low peak inspiratory pressure ventilation. MATERIALS AND METHODS: After saline lavage (10 cc/kg repeated as long as foamy retrieval was observed) two combined therapeutic strategies were examined: ventilation with high inspiratory pressures (35 cm H2O) with additional exogenous surfactant administration (100 mg/kg) and inhaled nitric oxide (10 PPM) (n=5, group 1) and low inspiratory pressure (20 cm H2O) ventilation under veno-venous ECMO support (n=5, group 2). The FiO2 was maintained at 1.0 in both groups. The paO2/FiO2 ratio was calculated in 30 minute intervals for 4 hours. After that the animals were sacrificed and the lungs examined macro- and microscopically. Aeration was described in a semiquantitative method using the alveolar expansion index. Oxygenation in group 1 was significantly better than in group 2, it increased significantly after surfactant but not after additional nitric oxide administration. However, the lungs in group 1 showed severe signs of baro/ergotrauma (Hyaline membranes, air leaks, infiltration of polymorphonuclear (PMN) granulocytes and macrophages, break down of alveolar capillary membranes) after 4 hrs of combined therapy, whereas the lungs in group 2 appeared normal. Adding surfactant and NO to a high tidal volume ventilation improved oxygenation, but did not prevent baro/ergotrauma. Ventilation with low inspiratory pressures combined with ECMO caused little baro/ergotrauma but adequate oxygenation could not be achieved, probably due to anatomical features of the rabbit which do not allow appropriate blood flow within the ECMO-circuit.

Animals↗

Bioelectronic noses: a status report. Part II.

The present state of the art to record or to mimic electronically the human senses of olfaction and taste is characterized. In this part II, strategies are outlined to utilize chemical and biological structures with their different complexities which serve as sensor elements in (bio-) electronic noses. Finally a survey is given on the computer-science aspects of odor recognition based on these elements.

Animals↗

Bioelectronic noses: a status report. Part I.

The present state of the art to record or mimic electronically the human senses of olfaction and taste is characterized. In this part I, an introduction to our present understanding in the development of electronic and bioelectronic noses is given. Finally the natural olfactory system is described in detail.

Brain↗

Biosensor development.

Current biosensor developments can be summarised by different trends. For traditional enzymatic biosensors such as glucose sensors, steady improvements of well known basic principles have been made in order to achieve better sensor stability. On the other hand, new affinity sensors such as nucleic acid sensors, transmembrane sensors, and sensors utilising whole cells or even cell networks have become of increasing interest. New ways to miniaturise biosensors and to control their interfaces down to the molecular level have been introduced (the bioelectronics approach). High-throughput screening based on various signal transduction principles has become of increasing importance.

Antigen-Antibody Reactions↗

Chiral discrimination of inhalation anesthetics and methyl propionates by thickness shear mode resonators: new insights into the mechanisms of enantioselectivity by cyclodextrins.

The discrimination of the enantiomers of methyl lactate, methyl 2-chloropropionate, and the inhalation anesthetics enflurane, isoflurane, and desflurane in the gas phase has been performed using thickness shear mode resonators. The selective coating was a modified perpentylated gamma-cyclodextrin derivative dissolved in a polysiloxane matrix. A new model for the sorption of the chiral compounds into the cyclodextrin cavities and into the polymer matrix was established for the purpose of characterizing the sensor responses. This characterization included the fitting of the sensor responses (preferential and nonpreferential sorption) according to the model and extracting the characteristic parameters. In particular we attempted to explain the observed variation of the chiral discrimination factor alpha with changing analyte or cyclodextrin concentrations and search for an invariable parameter, characteristic for a certain analyte-cyclodextrin combination. The process of chiral or "molecular" recognition was thoroughly investigated.

Anesthetics, Inhalation↗

Chiral discrimination using piezoelectric and optical gas sensors.

Odour perception in humans can sometimes discriminate different enantiomers of a chiral compound, such as limonene. Chiral discrimination represents one of the greatest challenges in attempts to devise selective and sensitive gas sensors. The importance of such discrimination for pharmacology is dear, as the physiological effect of enantiomers of drugs and other biologically active molecules may differ significantly. Here we describe two different sensor systems that are capable of recognizing different enantiomers and of qualitatively monitoring the enantiomeric composition of amino-acid derivatives and lactates in the gas phase. One sensor detects changes in mass, owing to binding of the compound being analysed (the 'analyte'), by thickness shear-mode resonance; the other detects changes in the thickness of a surface layer by reflectometric interference spectroscopys. Both devices use the two enantiomers of a chiral polymeric receptor, and offer rapid on-line detection of chiral species with high selectivity.

Alanine↗

Quartz crystal microbalances for quantitative biosensing and characterizing protein multilayers.

The use of quartz crystal microbalances (QCMs) for quantitative biosensing and characterization of protein multilayers is demonstrated in three case studies. Monolayers of QCM-based affinity biosensors were investigated first. Layers of a thiol-containing synthetic peptide constituting an epitope of the foot-and-mouse-disease virus were formed on gold electrodes via self-assembly. The binding of specific antibodies to epitope-modified gold electrodes was detected for different concentrations of antibody solutions. Oligolayers were studied in a second set of experiments. Dextran hydrogels were modified by thrombin inhibitors. The QCM response was used in a competitive binding assay to identify inhibitors for thrombin at different concentrations. Multilayers of proteins formed by self-assembly of a biotin-conjugate and streptavidin were investigated next. The QCM frequency response was monitored as a function of layer thickness up to 20 protein layers. A linear frequency decay was observed with increasing thickness. The decay per layer remained constant, thus indicating perfect mass coupling to the substrate. Frequency changes a factor of four higher were obtained in buffer solution as compared to measurements in dry air. This indicates a significant incorporation of water (75% weight) in the protein layers. This water behaves like a solid concerning the shear mode coupling to the substrate. The outlook discusses briefly the need for controlled molecular engineering of overlayers for subsequent QCM analysis, and the importance of an additional multiparameter analysis with other transducer principles and with additional techniques of interface analysis to characterize the mechanical coupling of overlayers as biosensor coatings. A promising trend concerns the use of QCM-arrays for screening experiments.

Biosensing Techniques↗

Odor, drug and toxin analysis with neuronal networks in vitro: extracellular array recording of network responses.

Neurons, by virtue of intrinsic electrophysiological mechanisms, represent transducers that report the dynamics of cell death, receptor-ligand interactions, alterations in metabolism, and generic membrane perforation processes. In cell culture, mammalian neurons form fault-tolerant, spontaneously active systems with great sensitivity to their chemical environment and generate response profiles that are often concentration- and substance-specific. Changes in action potential patterns are usually detected before morphological changes and cell damage occur, which provides sensitivity and reversibility. Such biological systems can be used to screen rapidly for novel pharmacological substances, toxic agents, and for the detection of certain odorants. Existing simple culture preparations can already be employed effectively for the detection of chemical compounds. So far, three strategies have been investigated in pilot experiments: (1) Substance-dependent major changes in spontaneous native activity patterns. All synaptically active agents (e.g. glutamate, strychnine, N-methyl D-aspartic acid) as well as metabolic poisons generate such changes. (2) Substance-dependent changes in network oscillations via disinhibition. The regularized, oscillatory activity is altered by synaptically and metabolically active substances, ion channel blockers, and toxins. (3) Detection of paroxysmal responses indicating major, pathological membrane currents in large subpopulation of cells. We have explored these three strategies via 64 channel array recordings using spontaneously active murine spinal cord cultures. The glycine receptor blocker strychnine reliably generated increased multichannel bursting at 5-20 nM and regular, coordinated bursting above 5 microM. During biculline-induced network oscillations many compounds alter oscillation frequencies or terminate activity in a substance-specific manner. Finally, the gp120 protein of the AIDS virus (at 1 microgram/ml) produces massive, unique paroxysmal discharges that may last as long as 2 min. These results indicate that cultured neuronal networks are practical systems that can be used for the detection and identification of a great variety of chemical substances. The concept of dynamic fingerprinting to identify specific compounds is discussed.

Animals↗

Characterization and optimization of microelectrode arrays for in vivo nerve signal recording and stimulation.

Revealing the complex signal-processing mechanisms and interconnection patterns of the nervous system has long been an intriguing puzzle. As a contribution to its understanding the optimization of the impedance behavior of implantable electrode arrays with via holes is discussed here. Peripheral axons will regenerate through these holes allowing for simultaneous nerve stimulation and signal recording. This approach is part of the ESPRIT project INTER and may eventually lead to devices driving sensory motor prosthesis with closed loop control. In the first set of experiments, micromachined platinum electrode arrays were prepared, characterized and optimized for nerve signal recording. The results of these studies are based on impedance spectroscopy and microscopic techniques. Equivalent circuits were modeled describing formally the electrical response behavior with ohmic resistances between 500 omega and 10 k omega. To attain low impedances for all electrodes on the INTER device, platinum from H2PtCl6 was electrodeposited, and sputter technology as well as electrochemical deposition from H2IrCl6 solution were used to produce thin iridium films. For the former, a lift-off process was established at one of the institutes to generate electrode structures with a line width of 5 microns. As a result in all three cases the electrodes showed almost constant impedances over the entire frequency range (10 Hz-1 kHz), which is relevant for nerve signal recording. In the second set of experiments, electrodes were optimized to allow for nerve stimulation. For this purpose, the charge delivery capacity (CDC) had to be increased and the impedance had to be decreased. Iridium oxide is the material of choice, because its CDC is much higher than the CDC of platinum at 75 microC/cm2 (Ziaie et al., 1991, IEEE Sensors & Actuators Transducers, 6, 124-127). A significant increase of the electrochemically active surface of the electrode structures could be observed by measuring the surface roughness. In first experiments, an activated iridium oxide film was formed with cyclic voltammetry and was evaluated using scanning force microscopy and impedance spectroscopy. The evaluation of the cyclic voltammograms showed a CDC up to 400 mC/cm2 for sputter deposited and oxidatively treated iridium films. Further investigations are directed towards increasing the stability of the iridium oxide electrodes with regard to long-term implants. Parallel experiments aim at the controlled axon adhesion without changing the impedance behavior of the described electrodes.

Electric Impedance↗