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Biomedical subjects

V Neuhoff

Publications and source records attributed to V Neuhoff.

At least 37 records · Page 2Linked to original sources
PubMed1987

Regional protein patterns of the embryonic chick brain: developmental shifts and the effects of light stimulation.

The developmental changes in the regional protein patterns of the embryonic brain of chick were analysed by micro-two-dimensional polyacrylamide gel electrophoresis stained by a highly sensitive procedure with Coomassie Brilliant Blue G-250 that completely avoids background staining. More than 400 protein spots were resolved in a computerized plot. Many proteins of the various embryonic brain regions varied with age. At least six of these proteins changed remarkably during development. One of them (Mr 14K) was prominent at 6 days and disappeared after 12 days of incubation. One acidic protein (Mr 30K) appeared at 12 days together with two acidic protein spots near tubulin and increased with development. A group of more basic proteins (Mr 94K) was prominent at 6 day incubation and decreased during development. One more basic protein (Mr 20K) was first observed at 18 days of incubation in the metencephalon. It was identified as myelin basic protein, and strikingly increased in all brain regions until hatching. A group of basic proteins with high molecular weight (Mr 94K) appeared at day 12 of incubation, and increased remarkably before hatching. They were identified to be collagen associated with brain capillary vessels. Stimulation by intermittent light in ovo from day 10 to day 16 caused a retarded appearance of these collagen proteins, and an increased formation of tubulin in the optic lobe of the developing chick embryo.

Animals↗
PubMed1987

[Facilitating cervix dilatation of the non-pregnant uterus by intracervical administration of gels containing prostaglandin and calcium chloride].

In a prospective, randomised study, 50 non-pregnant patients were treated intracervically with 3 ml 5% tylose, 50 micrograms sulprostone, 100 micrograms sulprostone gel, 3 ml 2.5 mM or 9.0 mM calcium chloride gel in order to soften the cervix 12-14 hours before diagnostic curettage. The gel was not used in a further 20 patients. To objectively demonstrate the priming effect, the force required for dilatation of the cervical canal was measured in Newtons, using a mechanical tonometer both before gel application and before the operation. In comparison with the administration of tylose only, the intracervical application of either sulprostone gel or calcium chloride gel led to a significant improvement in cervical dilatability. Tylose alone had a slight but measurable effect on the cervix. An increase in sulprostone from 50 micrograms to 100 micrograms or calcium molarity from 2.5 mM to 9.0 mM brought no further improvement in the dilatory effect. Dilatation-induced cervical lesions could be avoided by preoperative cervical ripening. After application of sulprostone, 3 out of 20 patients experienced doses-dependent uterine cramps, while all patients treated with calcium chloride gel were free of side effects. The intracervical administration of sulprostone and calcium chloride gel allowed gentle dilatation of the non-pregnant cervix, thus lowering the risk of uterine lesions. Under clinical aspects, cervical priming facilitates diagnostic and therapeutic procedures, which, in exceptional cases, can be performed without anaesthesia.

Administration, Intravaginal↗
PubMed1986

GABA neurones in retinas of different species and their postnatal development in situ and in culture in the rabbit retina.

The localisation of GABA immunoreactive neurones in retinas of a variety of animals was examined. Immunoreactivity was associated with specific populations of amacrine neurones in all species examined, viz. rat, rabbit, goldfish, frog, pigeon and guinea-pig. All species, with the exception of the frog, possessed immunoreactive perikarya in their retinal ganglion cell layers. These perikarya are probably displaced amacrine cells because GABA immunoreactivity was absent from the optic nerves and destruction of the rat optic nerve did not result in degeneration of these cells. GABA immunoreactivity was also associated with the outer plexiform layers of all the retinas studied; these processes are derived from GABA-positive horizontal cells in rat, rabbit, frog, pigeon and goldfish retinas, from bipolar-like cells in the frog, and probably from interplexiform cells in the guinea-pig retina. The development of GABA-positive neurones in the rabbit retina was also analysed. Immunoreactivity was clearly associated with subpopulations of amacrine and horizontal cells on the second postnatal day. The immunoreactivity at this stage is strong, and fairly well developed processes are apparent. The intensity of the immunoreactivity increases with development in the case of the amacrine cells. The immunoreactive neurones appear fully developed at about the 8th postnatal day, although the immunoreactivity in the inner plexiform layer becomes more dispersed as development proceeds. The immunoreactive horizontal cells become less apparent as development proceeds, but they can still be seen in the adult retina. The GABA immunoreactive cells in rabbit retinas can be maintained in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Fatty acid composition of myelin lipids from developing rat forebrain and spinal cord: influence of experimental hyperphenylalaninaemia.

The fatty acid composition of individual myelin lipids from rat forebrain and spinal cord was analysed at 20 and 30 days p.p. During this phase of rapid myelination the proportions of C 16:0 and C 18:0 fatty acids decreased whereas the relative amounts of long chain and unsaturated fatty acids increased in most lipid classes. This developmental shift was more pronounced in the forebrain and was different with respect to both magnitude and direction for each myelin lipid. The experimental induction of chronic hyperphenylalaninaemia (hyper-Phe) from day 3 p.p. lead to alterations in the rate of myelination, which were most pronounced in the forebrain. At 20 days, especially in the forebrain, chain elongation and desaturation of fatty acids were delayed. This delay was maximal in sphingomyelin fatty acids, C 18:0 and C 24:1. In hydroxycerebrosides, the shift in the fatty acid composition from C 16:0 to C 22:0 was accelerated in hyper-Phe rats. No significant difference in the fatty acid composition of any myelin lipid was found in the more mature myelin (spinal cord, 30 days p.p.). No evidence was found of a primary effect of hyper-Phe on myelination.

Animals↗
PubMed1986

Lysosomal protein degradation in experimental hyperphenylalaninaemia.

The influence of alpha-methylphenylalanine-induced hyperphenylalaninaemia (HYP) on the lysosomal protein degradation system in brain and liver of suckling rats was investigated. In both tissues cathepsin D and L activities, measured at 5, 10 and 15 days post partum (p.p.), exhibited no differences between experimental and control animals. N-Acetyl-beta-D-glucosaminidase (NAGase) activity in brain, measured at 10 and 15 days p.p., was not affected by HYP either. The release of valine and lysine from liver and brain homogenates respectively, serving as a measure for the lysosomal content of degradable proteins, was not influenced by HYP. Lysosomal integrity during incubation of homogenate was monitored by the recovery of NAGase activity in the cytosolic supernatant, and by the relative NAGase activity in total homogenates in the absence of the lysosome disrupting detergent Triton X-100. In conclusion, experimental HYP appears unlikely to influence the lysosomal protein degradation system in brain and liver of suckling rats.

Acetylglucosaminidase↗
PubMed1986

Induction of hyperphenylalaninemia in mice by ethionine and phenylalanine.

Female NMRI mice were fed diets containing l-ethionine (0.1 and 0.3% w/w) and phenylalanine (3% w/w), as well as respective control diets. Ethionine, the S-ethylated analog of methionine, was shown to inhibit phenylalanine hydroxylase in vivo, whereby in vitro kinetics remained unaffected. Treatment with ethionine resulted in fatty liver, reduced ATP content of liver, and alterations in serum amino acid concentrations. In the high dosage ethionine group, for instance, concentrations of Ala, Gly, Ser, Met, and Phe were increased whereas concentrations of Lys, Asp, and Pro were decreased. Applying ethionine together with phenylalanine resulted in hyperphenylalaninemia and phenylketonuria. Feeding phenylalanine alone also led to decreased activity of phenylalanine hydroxylase and increased concentration of Phe in serum. Ethionine only had a minimal effect on body weight gain; however, the hyperphenylalaninemic condition induced by application of the high dosage of ethionine and phenylalanine induced severe loss of body weight. A disturbed protein synthesis and protein phosphorylation might be the underlying mechanism of ethionine-induced suppression of phenylalanine hydroxylase.

Adenosine Triphosphate↗
PubMed1985

Amino acid depletion in the blood and brain tissue of hyperphenylalaninemic rats is abolished by the administration of additional lysine: a contribution to the understanding of the metabolic defects in phenylketonuria.

The elevated phenylalanine concentration in the blood of untreated phenylketonuric children is known to be paralleled by decreased concentrations of other amino acids in the blood and brain tissue. Due to the low availability of other large, neutral amino acids in the brain, protein synthesis in, and the normal development of, the brain are disturbed. A similar effect is observed in suckling rats rendered hyperphenylalaninemic by the daily injection of phenylalanine plus alpha-methylphenylalanine, an in vivo inhibitor of the phenylalanine-hydroxylating pathway in the liver. In this study, the simultaneous injection of lysine is shown to prevent the depletion of amino acids from the blood and brain tissue, and the retardation of brain growth, in suckling hyperphenylalaninemic rats. It is suggested that both amino acids, phenylalanine and lysine, are important rate-limiting substrates for the rapid protein anabolism of developing tissues. In the presence of an excess of phenylalanine, other amino acids, and in relation to its requirement during the phase of hyperplastic growth in particular lysine, are less available from the circulation and limit phenylalanine-stimulated protein synthesis in developing tissues. The supplementation of lysine to developing hyperphenylalaninemic rats prevents the consequences of this effect, i.e., the depletion of amino acids in the blood, and therefore, in the brain tissue, and the retardation of brain growth.

Age Factors↗
PubMed1985

Amino acid transport across the human blood-CSF barrier. An evaluation graph for amino acid concentrations in cerebrospinal fluid.

The correlation of cerebrospinal fluid (CSF)/serum concentration quotients was used as a method for identification of amino acids which are transported by a common carrier system across the blood-CSF barrier. Isoleucine, leucine, valine, phenylalanine, tyrosine and lysine were found to compete for the same carrier system. This group of amino acids in man was found to be different from the system described as a neutral amino acid carrier at the blood-brain barrier in rats. In man, methionine and tryptophan do not compete with the other neutral amino acids for the same carrier system. In contrast, lysine as a basic amino acid is found to be correlated with the same transport system as the five neutral amino acids. A graph for the evaluation of pathological amino acid concentrations in CSF is presented. Patients with a blood-CSF barrier dysfunction for proteins showed partly normal, partly increased, CSF/serum concentration quotients for the amino acids. Hydroxyproline could be identified as a constituent of the amino acid pool in CSF. For proline and hydroxyproline a special control system has to be suggested because of their smaller biological variance in CSF than in blood. Contrary to the other amino acids proline and hydroxyproline have a smaller biological variation in CSF than in serum.

Adolescent↗
PubMed1985

Glycerol phosphate dehydrogenase activity of oligodendrocytes isolated from adult pig brain: its inducibility by hydrocortisone.

Developing oligodendrocytes cultured in vitro express glycerol phosphate dehydrogenase (GPDH; EC 1.1.1.8) and are known to respond to glucocorticoid treatment by increased activity of GPDH. We present evidence that GPDH is enriched in white matter and oligodendrocytes of adult pig brain. Bulk-isolated oligodendrocytes maintained in culture for several weeks exhibit an almost constant level of GPDH activity. Furthermore, a 4-day stimulation with hydrocortisone induces GPDH specific activity of long-term cultured oligodendrocytes from adult pig brain.

Animals↗
PubMed1984

Localization of tyrosine-hydroxylase immunoreactive cells in rabbit retinal cultures.

Tyrosine-hydroxylase immunoreactivity is demonstrable by the indirect immunofluorescence technique in a small sub-population of retinal neurones cultured from 2-day-old rabbits. Staining was only weak, restricted to a few neurones, and could not be observed in cultures of less than 10 days. Autoradiographical analysis of [3H]dopamine uptake by retinal cultures strongly suggests that neurones containing tyrosine-hydroxylase immunoreactivity have the capacity to take up exogenous dopamine. These findings are in concordance with studies on rabbit retinal preparations.

Animals↗
PubMed1984

Isolation and cultivation of mature oligodendroglial cells.

CNS axons are ensheathed by myelin which is produced and maintained by oligodendrocytes. A disorder of this assembly results in functional disturbances, e.g., paralysis in multiple sclerosis. Methods are now available to isolate and cultivate oligodendrocytes in vitro. Thus, basic oligodendroglial properties can be now investigated: signals for oligodendroglial gene expression and their role in myelinogenesis and the interaction between oligodendrocytes and other neural cells by, e.g., the release of informational substances.

Animals↗
PubMed1984

Hereditary ocular dysproteinhydria of the aqueous humour with crystalline deposits.

We describe a family with coloured crystalline deposits on the corneal endothelium and on the lens surface. A protein with a molecular weight of about 110,000 and an isoelectric point of 7-8 could be demonstrated in the aqueous humour, which was not found in the controls or in the serum of the family members. The pedigree is suggestive of autosomal dominant inheritance. To our knowledge this is the first report of this entity.

Adult↗
PubMed1984

Bulk separation and long-term culture of oligodendrocytes from adult pig brain. I. Morphological studies.

A method is described by which oligodendrocytes from adult pig brains can be isolated. It results in a cellular preparation suitable for long-term culture. The entire procedure can be accomplished within 2-3 h. The purity of oligodendrocytes ranges between 80 and 95% depending on the Percoll gradient used and on the time in vitro. Yields between 2.5 and 4 X 10(7) cells per brain and plating efficiencies on the order of 60% make the system very useful for biochemical investigations. It was shown by immunocytochemical studies that oligodendrocytes produce extensive networks of processes, some of them having elaborate membranous expansions. Anti-galactocerebroside (GC) antibodies as well as anti-myelin basic protein (MBP), anti-Wolfgram protein (WP), anti-glial fibrillary acidic protein (GFAP), and monoclonal antibodies O1 and O4 are used to identify the cell types and to characterize the cellular composition of the cultures. Anti-GC and O1 are suitable markers for these oligodendrocytes. Both antibodies label similar cells, and the staining intensities are equally strong. In the case of O4, variable staining intensities are observed, and a few additional cells are labeled that are anti-GC-. After 3 1/2 weeks in culture, about 60% of the cells can be labeled by anti-MBP. Here too differences in staining intensities are observed. The anti-WP stain is too weak to be defined as positive. The percentage of GFAP+ cells lies in the range 15-20% at maximum. Cells were also mixed into collagen gels. This method appears to be more useful for outgrowth and branching of fibers than are monolayer systems. Drawbacks, however, include limited access for the antibodies and poor recovery of undamaged cells with their fibers.

Animals↗
PubMed1984

Bulk separation and long-term culture of oligodendrocytes from adult pig brain. II. Some biochemical data.

Oligodendroglial proteins labeled with radioactive amino acids were subjected to one- and two-dimensional polyacrylamide electrophoresis. Bands comigrating with myelin proteins, the basic protein (MBP), the proteolipid protein (PLP), and the Wolfgram protein (WP) doublet, were detected by Coomassie Blue staining and by autoradiography. The identity of the MBP and WP in the cellular material is evidenced by immunoblotting with specific antibodies. A comparative study of myelin samples from rat and pig CNS reveals that WP can be detected immunochemically in both species. Different protein patterns, however, are observed. Three protein bands are found with antibodies against the myelin-associated glycoprotein (MAG). The high-molecular-weight component prevails in pig myelin, whereas the medium-molecular-weight component is predominant in rat myelin. Moreover, two protein bands, of molecular weights 35,000 and 33,000 (Ol 1 and Ol 2), are present in high amounts in oligodendroglial particulate material but are not detectable in myelin. These oligodendroglial characteristic proteins are not species-specific, since they are found in preparations of cat oligodendrocytes as well. Activities of cerebroside sulfotransferase (EC 2.8.2.11) are low in freshly isolated cells and increase during the first week of culture. A reverse course of enzyme activities is observed with 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC 3.1.4.37). Values reach a minimum about day 5 in culture and recover their initial values. At day 10 they remain stable until the end of the third week of the culture period.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1984

Ammonia inhibits protein synthesis in slices from young rat brain.

Ammonium chloride, widely used as an inhibitor of lysosomal protein degradation, was shown to inhibit strongly protein synthesis in neocortical slices from 10-day-old rats at a concentration of 10 mmol/L. Its usefulness in experiments with brain tissue is doubted. The possible meaning of the finding with respect to ammonia toxicity is briefly discussed.

Alanine↗
PubMed1984

A "high-performance" 2D gel scanner.

We describe the design principles of a photometric flatbed scanner with a scan area of 250 X 250 mm2, a dynamic range of 4000 gray levels, a signal/noise ratio of 2000/1, a step size of 0.1 mm, and a step frequency of 2000 steps per second. It is controlled by a microcomputer and used to acquire data for quantitative evaluation of gels or chromatograms. The performance of the instrument is demonstrated by the data obtained by scanning gratings, gray step filters, and high-resolution electropherograms.

Electronic Data Processing↗
PubMed1984

Spot identification in two-dimensional patterns by a least-squares template matching.

The coordinates of the center of measured spots are nonlinearly transformed to get an optimal match between the transformed coordinates and the given coordinates of a reference pattern. The parameters of the transformation are determined by the minimum of a function of squared distances between all spots of the sample and of the reference pattern. The algorithm requires a priori defined correspondences between some pivot points in the sample and the reference and treats their distances differently from the others. The parameters of the transformation are the solutions of a system of nonlinear equations; their numerical values are obtained by iteration.

Amino Acids↗