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H Schulz

Publications and source records attributed to H Schulz.

At least 199 records · Page 11Linked to original sources
PubMed1994

Estimation of peroxisomal beta-oxidation in rat heart by a direct assay of acyl-CoA oxidase.

The contribution of peroxisomes to palmitate beta-oxidation in rat heart was estimated by either inhibiting mitochondrial beta-oxidation or measuring the activity of acyl-CoA oxidase. When respiratory inhibitors such as KCN or antimycin plus rotenone, or inhibitors of mitochondrial fatty acid uptake such as 2-tetradecylglycidic acid or 2-bromopalmitate, were used, degrees of inhibitions ranging from 24% to 87% were observed for palmitate beta-oxidation by a rat heart homogenate. Although the oxidation of palmitoyl-L-carnitine by coupled rat heart mitochondria was almost completely (94%) inhibited by KCN, the inhibition by antimycin plus rotenone was incomplete (77%) and was stimulated by L-carnitine. A direct assay of acyl-CoA oxidase, based on the spectrophotometric measurement at 300 nm of 2,4-decadienoyl-CoA formation from 4-trans-decenoyl-CoA, was evaluated with the aim of obtaining reliable values for the activity of this enzyme, which is presumed to catalyse the rate-limiting step of peroxisomal beta-oxidation. Activities determined by use of this assay were much higher than activities obtained by a coupled assay [Small, Burdett and Connock (1985) Biochem. J. 227, 205-210] commonly used to measure the activity of acyl-CoA oxidase. However, both methods yielded the same relative activities with different tissue homogenates. Based on an estimated palmitoyl-CoA oxidase activity of 0.3 nmol/min per mg of protein, the contribution of peroxisomes to palmitate beta-oxidation in a rat heart homogenate would optimally be 4%, and most likely is several-fold lower.

Acyl-CoA Oxidase↗
PubMed1994

CARE-LASS (calcein-release-assay), an improved fluorescence-based test system to measure cytotoxic T lymphocyte activity.

CARE-LASS is a highly sensitive, fast, simple and safe fluorometric microassay. Target cells are loaded with acetoxymethyl ester of calcein (calcein-AM) that passively crosses the cell membrane. Intracellular esterases convert the molecule to calcein, a polar fluorochrome which, in cells with intact plasma membranes, displays good retention characteristics and low pH sensitivity. In analogy to standard 51Cr release assays, the CARE-LASS system is based on the release of a marker into the supernatant that is measured by an automated fluorescence scanner and correlates with the number of lysed cells. We tested the CARE-LASS system by measuring cytotoxicity in major histocompatibility complex (MHC) class I and MHC class II restricted cytotoxic T lymphocyte (CTL) assays as well as lymphokine activated killer (LAK) mediated cytotoxicity. We applied a small set of target cell lines at various effector to target (E:T) ratios, at different antigen concentrations and compared CARE-LASS CTL data to data resulting from conventional 51Cr release assays. The CARE-LASS system provides a reliable and sensitive method to measure cell-mediated cytotoxicity.

Chromium↗
PubMed1994

Hepatic beta-oxidation of 3-phenylpropionic acid and the stereospecific dehydration of (R)- and (S)-3-hydroxy-3-phenylpropionyl-CoA by different enoyl-CoA hydratases.

The hepatic beta-oxidation of 3-phenylpropionic acid (PPA) was studied by the use of subcellular fractions and purified enzymes with the aim of characterizing intermediates and the subcellular location of this pathway. Respiration measurements with coupled rat liver mitochondria indicate that PPA is efficiently metabolized by mitochondrial beta-oxidation. In contrast, the peroxisomal beta-oxidation of this compound is at best a very slow process, as evidenced by the low activity of peroxisomal acyl-CoA oxidase toward 3-phenylpropionyl-CoA. In mitochondria, 3-phenylpropionyl-CoA is effectively dehydrogenated to cinnamoyl-CoA, which is only slowly converted to benzoylacetyl-CoA due to the unfavorable equilibrium of the hydration of cinnamoyl-CoA to 3-hydroxy-3-phenylpropionyl-CoA. Benzoylacetyl-CoA is a substrate of 3-ketoacyl-CoA thiolase. The dehydration of 3-hydroxy-3-phenylpropionyl-CoA to cinnamoyl-CoA forms the basis for a sensitive and stereospecific assay of enoyl-CoA hydratases. The progress of this reaction, which proceeds to near completion, can be measured spectrophotometrically at 308 nm. Soluble mitochondrial and peroxisomal enoyl-CoA hydratases only act on the (R,L) isomer, whereas the peroxisomal D-3-hydroxyacyl-CoA dehydratase is specific for the (S,D) isomer. Both substrates can be easily prepared from the commercially available enantiomeric acids. It is concluded that PPA, a key compound in Knopp's classical study that led him to formulate the principle of beta-oxidation, is overwhelmingly, if not completely, degraded by mitochondrial beta-oxidation.

Acyl Coenzyme A↗
PubMed1994

Primary structure of the large subunit of trifunctional beta-oxidation complex from pig heart mitochondria.

The amino-terminal and internal sequences of the isolated large subunit of trifunctional beta-oxidation complex from pig heart mitochondria were determined by Edman degradation. The results demonstrated that the sequence of this novel beta-oxidation enzyme is identical with the sequence recently reported for a porcine gastrin binding protein that serves as the gastrin receptor on parietal cell surfaces. Evidence is provided to show that it is unlikely that the porcine gastrin binding protein has such a sequence. The data lead us to conclude that the mature large subunit of porcine trifunctional beta-oxidation complex is composed of 727 amino acid residues with a calculated molecular weight of 79,113, while the precursor of this long-chain fatty acid oxidation enzyme has a mitochondrial presequence consisting of 36 residues and a calculated M(r) of 83,099.

Amino Acid Sequence↗
PubMed1994

Delta 3,5, delta 2,4-dienoyl-CoA isomerase from rat liver mitochondria. Purification and characterization of a new enzyme involved in the beta-oxidation of unsaturated fatty acids.

Mitochondrial delta 3,5, delta 2,4-dienoyl-CoA isomerase, which catalyzes the conversion of 3,5-octadienoyl-CoA to 2,4-octadienoyl-CoA, was purified from rat liver 370-fold at almost 30% yield by a six-step purification procedure. The final preparation appeared to be homogeneous as judged by gel electrophoresis. The molecular weights of the native enzyme and its subunit(s) were estimated to be 126,000 and 32,000, respectively. The purification of delta 3,5, delta 2,4-dienoyl-CoA isomerase completes the characterization of the enzymes functioning in the NADPH-dependent pathway for the beta-oxidation of unsaturated fatty acids with double bonds extending from odd-numbered carbon atoms. This novel pathway may not be operative in peroxisomes because delta 3,5, delta 2,4-dienoyl-CoA isomerase was only detected in mitochondria. Substrates of this pathway are 2,5-dienoyl-CoAs formed from 5-enoyl-CoAs by acyl-CoA dehydrogenase. Two sequential isomerization reactions catalyzed by delta 3, delta 2-enoyl-CoAs isomerase and delta 3,5, delta 2,4-dienoyl-CoA isomerase, respectively, convert 2,5-dienoyl-CoAs to 2,4-dienoyl-CoAs, which are reduced by NADPH-dependent 2,4-dienoyl-CoA reductase (EC 1.3.1.34) before reentering the beta-oxidation spiral.

Animals↗
PubMed1994

Spectrophotometric assay of 2,4-dienoyl coenzyme A reductase with 5-phenyl-2,4-pentadienoyl-coenzyme A as substrate.

The spectrophotometric assay of 2,4-dienoyl coenzyme A (CoA) reductase (EC 1.1.1.34) was modified to improve the linearity and sensitivity of this method. 5-Phenyl-2,4-pentadienoyl-CoA, which has an absorbance maximum at 340 nm with an extinction coefficient of 44,300 M-1 cm-1, was synthesized and used as substrate. This compound is reduced by nicotinamide adenine dinucleotide phosphate (NADPH)-dependent 2,4-dienoyl-CoA reductase to 5-phenyl-3-pentenoyl-CoA. When a tissue homogenate serves as an enzyme source, the product is further metabolized by delta 3, delta 2-enoyl-CoA isomerase (EC 5.3.3.8) to 5-phenyl-2-pentenoyl-CoA, which is hydrated to 5-phenyl-3-hydroxypentanoyl-CoA by enoyl-CoA hydratase (EC 4.2.1.17). The modified assay method, which measures the decrease in absorbance at 340 nm due to the reduction of 5-phenyl-2,4-pentadienoyl-CoA and the oxidation of NADPH, is linear for a longer period of time and is twice as sensitive as the conventional assay with 2,4-decadienoyl-CoA as substrate.

Acyl Coenzyme A↗
PubMed1994

[Malignant melanoma in epiluminescent microscopy].

Comparison by skin epiluminescent microscopy of 60 malignant melanomas and 130 junctional and compound naevocellular naevi revealed statistically significant differences for 12 characteristics. The following microscopical in vivo signs that can be defined by histopathological examination were found only in malignant or premalignant melanocytic lesions: digitated radial streaming, greyish-blue reticular patterns, opaque gypseous alabaster lacunas, brown/black dots on a blue/greyish background, pseudopodia at the periphery and blue-in-pink areas. Nevertheless, specific epiluminescent phenomena of histopathologically defined malignant melanomas are still unknown. Therefore, diagnostic evaluation of malignancy is based mainly on microscopic assessment of combined details of morphology and colour of melanocytic tumours.

Cell Transformation, Neoplastic↗
PubMed1994

The relationship between mitochondrial activation and toxicity of some substituted carboxylic acids.

The activation of 4-bromocrotonic acid, 4-bromo-2-octenoic acid, valproic acid, and 3-methylglycidic acid by conversion to their CoA thioesters and the effects of these carboxylic acids on palmitoylcarnitine-supported respiration were studied with rat liver and rat heart mitochondria. 4-Bromocrotonic acid was activated by both liver and heart mitochondria, whereas 4-bromo-2-octenoic acid and valproic acid were only activated by liver mitochondria. 3-Methylglycidic acid was not a substrate of mitochondrial activation. All of the carboxylic acids that were activated also inhibited palmitoylcarnitine-supported respiration. 3-Methylglycidoyl-CoA was found to irreversibly inhibit 3-ketoacyl-CoA thiolase in a concentration-dependent and time-dependent manner. Together, these results lead to the conclusion that substituted medium-chain carboxylic acids, which enter mitochondria directly, may inhibit beta-oxidation as long as they are activated and perhaps further metabolized in the mitochondrial matrix to compounds that sequester CoA and/or inhibit beta-oxidation enzymes. Liver is more susceptible to inhibition by such xenobiotic carboxylic acids due to the broader substrate specificity of its mitochondrial medium-chain acyl-CoA synthetase (EC 6.2.1.2).

Animals↗
PubMed1994

The effect of valerian extract on sleep polygraphy in poor sleepers: a pilot study.

The effect of acute and repeated treatment (seven days) with a valerian extract (Valdispert forte, 405 mg t.i.d.) on objective and subjective measures of sleep was studied. Polysomnography was conducted in 14 elderly poor sleepers on three nights, at one-week intervals (N0, N1, N2). N0 was an adaptation night, N1 and N2 the first and last night under treatment. Six subjects received placebo and eight subjects valerian. Subjects in the valerian group showed an increase in slow-wave sleep (SWS) and a decrease in sleep stage 1. Density of K-complexes was increased under active treatment. There was no effect on sleep onset time or time awake after sleep onset. REM sleep was unaltered. There was also no effect on self-rated sleep quality. We hypothesize that valerian increases SWS in subjects with low baseline values.

Aged↗
PubMed1994

BMS-181168 for protection of the human brain against hypoxia: double-blind, placebo-controlled EEG mapping studies.

In a double-blind, placebo-controlled, cross-over trial, the antihypoxidotic properties of BMS-181168 (previously BMY 21502)--a 1-[[1-[2-(trifluoromethyl)-4-pyrimidinyl]-4-piperidinyl]methyl]-2- pyrrolidinone alleviating impairment of learning and memory in the animal--were studied utilizing EEG mapping under an experimental hypoxic hypoxidosis. The latter was induced by a fixed gas combination of 9.8% oxygen (O2) and 90.2% nitrogen (N2) (found at 6000 m altitude), which was inhaled for 23 minutes under normobaric conditions by 16 healthy male volunteers (aged 23-35 years, mean 27.2 years). After an adaptation session, they received in randomized order at weekly intervals oral single doses of placebo, or of 100 mg, 200 mg, and 400 mg BMS-181168. Evaluation of blood gases (PO2, PCO2, SO2), adverse events, and EEG mapping was carried out prior to drug administration and 2, 4, 6 and 8 hours post-drug, on each occasion under normoxic and transient hypoxic conditions. Hypoxemia was controlled by drawing arterialized capillary blood samples from the earlobes after hyperemization of the latter (after 0, 14, and 23 minutes of hypoxic gas inhalation) and by oximetry. After 23 minutes of inhalation, analysis showed a drop in PO2 from 98 to 48 mm Hg, in PCO2 from 41 to 31 mm Hg, and in SO2 from 97 to 80%. Descriptive statistical analyses of EEG mapping data demonstrated under hypoxia/placebo conditions an increase in delta/theta activity and a decrease in alpha activity as well as a slowing of the delta/theta centroid and an increase in the alpha and beta centroid, which suggests a marked deterioration in physiological vigilance.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

[Functional orthoses in insufficiency of the anterior cruciate ligament. Biomechanical comparative studies in vivo and with a model].

UNLABELLED: We tested 4 functional braces for anterior cruciate ligament insufficiency (CTI, Lennox Hill, Ultratech, Proshifter) in vitro and in vivo. In vitro all braces limited the anterior drawer. The maximal anterior drawer was 3.08 mm (CTI), 5.16 mm (Ultratech), 6.25 mm (Lennox-Hill), 7.33 mm (Proshifter). In vivo anterior drawer was controlled at low shear loads (67 N). With high shear loads the anterior drawer was significantly reduced but the tibiofemoral translation was significantly reduced but the tibiofemoral translation was higher than in the uninjured knee. Without brace: 16.25 mm; CTI: 10.15 mm; Ultratech: 12.10 mm; Lennox-Hill: 9.80 mm; Proshifter: 12.75 mm; CONTROL: 8.05 mm. The active anterior drawer was reduced but not normalized by all braces. Without brace: 8.75 mm; CTI: 5.35 mm; Ultratech: 5.6 mm; Lennox-Hill: 5.45 mm; Proshifter: 6.75 mm; CONTROL: 4.25 mm. None of the braces tested could control the rotation efficiently.

Anterior Cruciate Ligament↗
PubMed1994

Regulation of fatty acid oxidation in heart.

The control of fatty acid oxidation in heart is reviewed with special emphasis on the energy-linked regulation of this process. Studies with perfused working hearts and isolated mitochondria have revealed an inverse relationship between the energy-dependent rate of fatty acid oxidation and the intramitochondrial ratio of [acetyl-CoA]:[free CoA] at sufficiently high concentrations of fatty acids. Studies with isolated enzymes demonstrated a strong inhibition of 3-ketoacyl-CoA thiolase by acetyl-CoA at low concentrations of free CoA. Together these observations prompted the proposal that the rate of fatty acid oxidation is tuned to the energy demand of heart via the regulation of 3-ketoacyl-CoA thiolase by the [acetyl-CoA]:[free CoA] ratio. Evidence in support of this regulatory model has been obtained with isolated rat heart mitochondria in which either the activity of 3-ketoacyl-CoA thiolase was decreased by use of mechanism-based inhibitors or the intramitochondrial ratio of [acetyl-CoA]:[free CoA] was adjusted with L-carnitine. Because intermediates of beta-oxidation normally do not accumulate in mitochondria, it remains unclear how the entry of fatty acyl-CoA into the beta-oxidation spiral is tuned to the activity of 3-ketoacyl-CoA thiolase. A control of fatty acid oxidation in heart via the regulation of carnitine palmitoyltransferase I by malonyl-CoA has not been established even though malonyl-CoA is present in this tissue and strongly inhibits myocardial carnitine palmitoyltransferase I.

Animals↗