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Biomedical subjects

G Weidemann

Publications and source records attributed to G Weidemann.

At least 19 recordsLinked to original sources

Transfer and counterconditioning of conditional control in the rabbit nictitating membrane response.

Two experiments using the rabbit nictitating membrane response investigated whether training in one conditional discrimination (A-->X+, B-->X-), enabled the feature cues (A and B) to modulate responding to another CS (Y) trained as a target stimulus in a second conditional discrimination (C-->Y+, D-->Y-). There was near-complete transfer of the feature cue's conditional control, indicating that the feature cue's ability to modulate responding is not based on an association specific to the training target. Experiment 2 also revealed that the role of a stimulus to act as a conditional cue is affected by its ability to act as a simple conditioned excitor or inhibitor. Following initial acquisition of two conditional discriminations, two feature cues were reinforced in a pattern consistent with the initial conditional discrimination (A-->+, B-->-), where as the other two feature cues were reinforced in the reverse pattern to that of the original conditional discrimination (C-->-, D-->+). Subsequent tests revealed that the reversed training of the feature cues interfered with the original conditional discriminations. The results are consistent with theories that the feature cue gains an association with a representation of the emotional attributes of the US, which acts to modulate responding to the target stimulus through a diffuse change in motivational level. However, hierarchical theories of conditional discriminations that assume a lack of CS-specificity may also be able to explain the findings.

Analysis of Variance↗

Long-range tilt orientational order in phospholipid monolayers: a comparative study.

Monolayers of dipalmitoyl-phosphatidyl-N-monomethylethanolamine (DPP(Me)E) and dipalmitoyl-phosphatidyl-N,N-dimethylethanolamine (DPP(Me2)E) are studied and compared with dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), and dipalmitoyl-phosphatidylcholine (DPPC) to characterize the influence of the headgroup size. The properties of the condensed phases of DPP(Me2)E and DPP(Me)E are between those of DPPC and DMPE or DPPE. DPPC domains are elongated and the orientation changes continuously, whereas DMPE domains are compact and the orientation jumps at curved lines. The domains of DPP(Me2)E and DPP(Me)E are compact, and not elongated. The orientation changes continuously by 360 degrees around a point in the centered domains, and jumps of the orientation occur only in the case of twinning. Furthermore, the size of the headgroup influences the erection of the aliphatic chains. For DPPC and (DPP(Me2)E), no complete erection of chains occurs, whereas for DPP(Me)E the surface pressure required for the complete erection of chains is much higher than for DPPE. The same tendency is found for the collapse. DPPC monolayers do not collapse. DPP(Me2)E monolayers collapse at a much higher surface pressure than those of DPP(Me)E and DPPE.

1,2-Dipalmitoylphosphatidylcholine↗

Determination of potassium by dry reagent carrier technology: a multicentre evaluation.

We describe the construction, the reaction principle and the performance of Reflotron K+, a new Reflotron test for the quantitative determination of potassium in serum and heparinized plasma. The reaction principle is based on the introduction of the potassium cation via valinomycin into a non-polar phase; the accompanying loss of protons from the non-polar phase is detected by the colour change of a pH indicator. The multicentre evaluation of the reagent carrier system showed in median CVs of < 0.9% (within-series in heparinized plasmas) and 1.3% (run-to-run in control sera). The recovery in control sera was +/- 4% for seven laboratories. In the method comparison with flame emission spectrometry, using sera and heparinized plasma samples, regression analysis yielded correlations with slopes of 1.00 +/- 0.04 (median slope 1.005) and negligible intercepts. The reagent carrier system showed a linear response in the measuring range 2-12 mmol/l. Bilirubin (up to 513 mumol/l), triacylglycerols (up to 5,7 mmol/l), sodium (135-189 mmol/l) and ammonium ions (up to 590 mumol/l) did not interfere with the test. Comparison with the results from flame atomic emission spectrometry shows that the recoveries of Reflotron K+ and the direct potentiometric method are slightly and similarly influenced by total protein. With a panel of 28 drugs tested, no interference could be detected. Reflotron K+ provides a precise and reliable procedure for the measurement of potassium in serum and heparinized plasma.

Chemistry, Clinical↗

Development and evaluation of a reagent carrier with a new reaction sequence for the determination of creatinine in blood, plasma, serum and urine.

After a short outline of the history of creatinine determination methods we describe the development of a dry-reagent-carrier system for the reflometric determination of the creatinine concentration in blood, plasma, serum and urine (Reflotron Creatinine (new)). The method is based on a sequence of enzymatically catalyzed reactions producing H2O2, but which in contrast to the previously used procedure do not lead to the formation of creatine as an intermediate. Hence, pretreatment of sample material to eliminate endogenous creatine is no longer necessary. In the indicator reaction, use is made of an imidazole derivative as the chromogen. The dye formed in the presence of peroxidase can be measured by reflectance photometry beyond the long-wave absorption bands of haemoglobin and bilirubin at 642 nm. We present in detail the results of the multicentre evaluation of the analytical properties of this new test principle. The data obtained show that Reflotron Creatinine (new) correlates well with the routine method Creatinine PAP, which was used as a comparison method, with respect to accuracy and precision and even surpasses it with respect to specificity. Advantages over the first generation of Reflotron Creatinine are: shorter reaction time, longer stability of the reagent carrier, no interference by bilirubin and reduced interference by haemoglobin.

Blood Chemical Analysis↗

A baseline-ecosystem approach to the analysis of ecotoxicological effects.

Since 1980 the German Ministry of Research has sponsored a number of ecotoxicological research projects aimed at the analysis of effects of environmental chemicals on terrestrial ecosystems. Some of these field studies were conducted by using a baseline-ecosystem approach, the theoretical framework of which is given. It is demonstrated by the baseline ecosystem derived from a ruderal site. The results from the German ecotoxicology program concerning some features of structural and functional responses of ecosystems to chemicals are summarized. Many effects observed in "real nature" could be attributed to the substances under study as side effects due to the integrative approach. This means that without knowledge of the structure and function of a biogeocoenosis reliable predictions of chemically induced changes at the ecosystem level will not be possible.

Ecology↗

Alpha-amylase determination with the Reflotron reagent carrier system: use of whole blood, plasma, and serum, and effect of isoenzymes.

In the Reflotron Amylase dry-reagent carrier system (Boehringer Mannheim GmbH) a new substrate is used for determining total amylase (EC 3.2.1.1) activity:indolyl-alpha-D-maltoheptaoside. The procedure shows low imprecision (median CV less than 3.2%), and results for sera, plasma, and capillary and venous blood (y) correlate well with those of a conventional alpha-amylase method involving p-nitrophenyl (PNP)-maltoheptaoside substrate (x) (for 209 blood samples: y = 0.981x + 9.7; r = 0.994). Correlation was also excellent with a method involving maltotetraose as substrate (r = 0.987). Attachment of an indoxyl residue rather than a PNP group to the maltoheptaoside did not affect the substrate response to pancreatic or salivary isoenzyme activity. Therefore, the relative proportion of these isoenzymes did not affect the correlation between the Reflotron Amylase reagent carrier and the alpha-amylase PNP-maltoheptaoside method. With a reaction time of less than 3 min, this system is especially suitable for amylase determination in situations where a prompt result is required.

Humans↗

Quantitative determination of valproic acid by means of gas chromatographic headspace analysis.

We have developed a simple but accurate procedure to monitor, by means of gas chromatographic headspace analysis, the concentration of valproic acid (VPA) in serum. Unlike previously described gas chromatographic methods, a preanalytical separation step is unnecessary. Irreversible sample loss, a pitfall in gas chromatographic extraction procedures caused by evaporation during the extraction process of this relatively volatile substance, is avoided. After acidifying and adding internal standard (2- ethylbutanoic acid), the serum sample is sealed hermetically in a special headspace tube. After heating, a vapour phase fraction is injected into the chromatographic column. This method reduces sample preparation time to a few minutes. Analysis time is only three minutes, and the method can easily be automated. Contamination of chromatographic columns by higher molecular mass serum ingredients is avoided.

Chromatography, Gas↗

A sensitive dual wavelength microspectrophotometer for the measurement of tissue fluorescence and reflectance.

The construction of a microscope photometer using prefabricated elements is described. To illuminate the tissue, a Leitz Ultropac is applied. To enlarge the wavelength range, its illuminating glass lens is replaced by an Acryl glass zonal lens. Two separate light channels with separate lamps, monochromators and photomultipliers allow the measurement of fluorescence excitation and emission spectra as well as of reflection spectra. By chopping the light, light pulses and dark currents are measured 8.33 times a second. By an integration circuit the signal-to-noise ratio for small signals is improved. The instrument allows us to detect the increase of 4 ng/ml NADH (solution of pH 7.38) in an area of 0.2 mm2.

Brain Chemistry↗

Separation of glutathione reductase in human serum by gradient gel electrophoresis.

A system of high resolution polyacrylamide gel electrophoresis is introduced. It allows the rapid separation of proteins from serum and tissue homogenates under non-denaturating conditions, and proves especially suited for the comparative analysis of enzymes. The enzyme glutathione reductase is identified on the gradient gels by contact printing with agar substrate gels, and the multiple forms of glutathione reductase in serum are demonstrated.

Electrophoresis, Polyacrylamide Gel↗

[Light sensitivity of creatine kinase in control sera].

The light sensitivity of creatine kinase in different control sera was investigated. Control sera were dispensed into reaction vessels, then exposed for 4 h at 25 degrees C to a light source equivalent in intensity to normal laboratory illumination. In 12 out of 22 control sera, the catalytic concentration of the creatine kinase fell by 25-63%. Under the same conditions, in patient sera, creatine kinase is stable. In the "light sensitive" control sera, the instability of the enzyme increased with the intensity of illumination. When exposed to sunlight creatine kinase was unstable in all the investigated control and patient sera. The addition of mercaptoethanol stabilized the creatine kinase in "light sensitive" control sera. The reason for the different light sensitivity of different control sera is not known.

Creatine Kinase↗

[Interference by glutathione reductase in the "CK-MB-inhibition-test": elimination of the catalytic activity of glutathione reductase by anti-glutathione reductase (author's transl)].

Glutathione reductase interferes in the determiniation of creatine kinase-MB by the "CK-MB-inhibition-test". In 36 out of 55 serum samples with normal and elevated catalytic activities of creatine kinase, "negative" values greater than 14 U/l were obtained for the creatine kinase-MB. This interference by glutathione reductase is avoided by preincubation of the serum samples for 5 min with anti-glutathione reductase.

Catalysis↗

[Multiple forms of NADPH-dependent glutathione reductase in serum. Studies on the NADPH-dependent glutathione-reductase in serum II. (author's transl)].

Sera with elevated activities of glutathione reductase were investigated by gel electrophoresis in agar or polyacrylamide, and by gel filtration. In both separation methods the glutathione reductase activity in individual samples was resolved, showing up to three fractions differing in rate of migration and molecular size. The fractions with the lowest and highest molecular weight, corresponded respectively to the slowest and fastest migrating bands in agar gel electrophoresis. Preincubation of the serum samples with FAD or neuraminidase had no effect on the rate of migration of the three fractions. After the addition of beta-mercaptoethanol to the serum, gel electrophoresis and gel filtration showed only the enzyme fraction with the slowest rate of migration and the lowest molecular weight (140,000). The other two fractions reappeared after removal of the thiol from the serum. Further studies on the isolated (agar gel electrophoresis) fractions showed the existence of oligomeric forms of the enzyme, which are reversibly interconvertible.

Electrophoresis, Agar Gel↗

[New method for intranodular administration of 2,4-dinitro-1-chlorobenzene].

2,4-Dinitro-1-chlorobenzene (DNCB) proved to be an excellent stimulator for immunotherapy of melanom. Because of its poor solubility the therapy is limited to topical application. Our investigations have shown that an aqueous, injectable solution may be obtained by using dimethylsulfoxide (DMSO) that permits solubilization.

Adjuvants, Pharmaceutic↗

[Optimized determination and properties of NADPH-dependent glutathione reductase in serum. Studies on serum glutathione reductase, I. (author's transl)].

Reaction conditions were optimized for the determination of serum glutathione reductase, which has not yet been investigated systematically. Imidazole was found to be the most suitable buffer material; the highest glutathione reductase activity in serum was always obtained with imidazole/HCl buffer, which, in contrast to all other tested buffers, also resulted in the maximal enzyme activity without preincubation. In imidazole buffer, the pH-activity curve of serum glutathione reductase shows a broad optimum between pH 6.5 and 6.9. A GSSG concentration of 2 mmol/l and a NADPH concentration of 0.43 mmol/l gave maximal enzyme activity and a linear reaction over 10 min up to 20 U/l test solution. An investigation of serum glutathione reductase activity from 100 clinically healthy probands gave values between 20 and 50 U/l. In the optimized assay system the glutathione reductase in the serum reacts specifically with GSSG and NADPH.

Autoanalysis↗