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Biomedical subjects

E Koller

Publications and source records attributed to E Koller.

At least 19 recordsLinked to original sources
PubMed1992

Characterization of the chicken alpha 1(VI) collagen promoter.

The promoter of the chicken alpha 1(VI) collagen gene resembles the 5'-flanking regions of many housekeeping genes. It lacks a canonical TATAA box but contains potential binding sites for transcription factors AP1 and SP1. The promoter region has a relatively high GC content and forms a typical CpG island. In accordance with the absence of a TATAA element, the gene contains multiple transcription-initiation sites distributed over 80 bp genomic DNA. A 621-bp fragment derived from the 5' end of the alpha 1(VI) collagen gene is able to direct transcription of a heterologous reporter gene in transient-expression assays. Other DNA fragments that are either shorter or longer than the 621-bp fragment show markedly reduced promoter activity. Thus, the basic promoter element of the alpha 1(VI) collagen gene must reside within this 621-bp fragment.

Animals↗
PubMed1992

Structural comparison of the chicken genes for alpha 1(VI) and alpha 2(VI) collagen.

The chicken alpha 1(VI) polypeptide is encoded by a single gene spanning 21 kbp of genomic DNA. This gene is composed of 34 exons and 33 introns. Its structure is closely related to that of the alpha 2(VI) collagen gene, suggesting that the two genes evolved by gene duplication. Both genes contain 19 exons coding for the triple-helical domain. These exons are multiples of 9 bp (27, 36, 45, 54, 63 and 90 bp) and encode an integral number of collagenous Gly-Xaa-Yaa triplets. Since there is no convincing correlation to a building block of 54 bp, it is unlikely that type VI collagen has evolved from a primordial 54-bp module as suggested for all fibrillar collagens.

Amino Acid Sequence↗
PubMed1992

Binding of 111In-labeled HDL to platelets from normolipemic volunteers and patients with heterozygous familial hypercholesterolemia.

High density lipoproteins (HDLs; d = 1.063 - 1.21 g/ml) were isolated by ultracentrifugation and radiolabeled with 111In. The in vitro binding onto platelets from healthy volunteers (n = 15) and patients (n = 36) with heterozygous familial hypercholesterolemia (FH) was investigated. Binding was saturable and indicated high-affinity binding sites, which bound 1,882 +/- 361 ng protein of 111In-HDL/10(9) platelets (dissociation constant [Kd] = 7 +/- 3 micrograms protein/ml) in healthy volunteers and significantly (p less than 0.01) lower amounts in the FH patients (1,012 +/- 439 ng protein of 111In-HDL/10(9) platelets [Kd = 12 +/- 4 micrograms protein/ml]; p less than 0.01). The capacity to displace one half of the bound ligand (IC50) amounted to 14 +/- 3 micrograms protein/ml in healthy volunteers and 22 +/- 9 micrograms protein/ml in FH patients (p less than 0.001). Treatment with lipid-lowering drugs (gemfibrozil, alone or in combination with cholestyramine) in 10 patients resulted in an increased HDL binding capacity: before treatment, 1,280 +/- 883; after 2 months of treatment, 2,052 +/- 873 (p less than 0.05); and after 6 months of treatment, 2,127 +/- 812 ng protein/10(9) platelets (p less than 0.01). There was a significant (p less than 0.001) correlation between 111In-HDL binding data and plasmatic lipid and lipoprotein values. Furthermore, those FH patients with the additional risk factors of smoking (p less than 0.05) and hypertension (p less than 0.01) showed significantly lower 111In-HDL binding onto platelets. The findings indicate specific 111In-HDL binding sites for human platelets, which may be decreased in patients with heterozygous FH. Upregulation of HDL binding sites during lipid-lowering medication therapy supports the hypothesis that high-affinity HDL binding is involved in hyperlipemic disorders and is possibly related to the reactivity of platelets.

Adult↗
PubMed1991

The promoter of the chicken alpha 2(VI) collagen gene has features characteristic of house-keeping genes and of proto-oncogenes.

The promoter of the chicken alpha 2(VI) collagen gene reveals several interesting features characteristic of house-keeping genes and growth control related genes. It does not possess a typical TATAA or CAAT box, but it contains several potential binding sites for transcription factors Sp1 and ETF. The 5' flanking region of the gene forms a typical 'CpG island' where the dinucleotide sequence CpG occurs with high frequency relative to the bulk genome. Consistent with the lack of a TATAA element, the gene contains multiple transcription initiation sites distributed over 75 bp of genomic DNA. A short DNA fragment (207 bp) encompassing all the transcription initiation sites and the entire CpG island shows strong promoter activity when linked to a heterologous reporter gene. The upstream region of the promoter harbours a long homopurine/homopyrimidine element (403 bp) which is sensitive to endonuclease S1. This element might have the ability to adopt an intramolecular hairpin triplex structure and could play a role in the organization of the chromatin at the alpha 2(VI) collagen locus. Our results demonstrate that the structure of the alpha 2(VI) collagen promoter is completely different from that of any other collagen promoter characterized so far.

Animals↗
PubMed1991

[Pathologico-anatomic characteristics of conservatively treated breast carcinomas in Basel].

Conservation treatment was applied to 406 patients in Basel, between 1977 and 1986, who accounted for 25% of all cases of mammary carcinoma. Patients with favourable prognostic factors had been selected for this therapeutic approach. The primary tumor bed was re-excised in about 50% of all operations. Intramammary recurrences were recorded from 9% of all cases, through an average follow-up period of 53 months. Presence of carcinoma residues in re-excised specimens was an important risk factor for development of locally delimited recurrence. Additional risks were relating to presence of tumor residues in final excision margins as well as to invasive lobular carcinoma. No combination of histopathological risk factors has so far become known as an absolute contraindication to conservation treatment of mammary carcinoma.

Adult↗
PubMed1991

Indium-111-labeled low-density lipoprotein binds with higher affinity to the human liver as compared to iodine-123-low-density-labeled lipoprotein.

The interaction of 111In-low-density lipoprotein (LDL) and 123I-LDL with human liver-plasma membranes was investigated and compared. LDLs were isolated by sequential ultracentrifugation and radiolabeled either with 123I (using lodogen or iodine-monochloride) each followed by purification with gel-chromatography or dialysis) or 111In (using cyclic DTPA-anhydride). LDL concentrations of 0.1 to 32 micrograms protein/ml were used for direct binding assays investigating the specific binding of labeled LDL (in the presence of a 50-fold excess of unlabeled LDL) to human liver apoB-receptors. In separate experiments, displacement of bound 111In-(123I)-LDL by unlabeled LDL was studied. Human liver plasma membranes bound 239 +/- 26 ng protein of 111In-LDL/mg protein and 148 +/- 18 ng protein of 123I-LDL/mg protein specifically (p less than 0.001). The corresponding dissociation constants were 0.6 +/- 0.2 and 1.2 +/- 0.7 micrograms protein/ml, respectively (p less than 0.001). The capacity of unlabeled LDL to displace bound 111In-LDL was four times higher than that for 123I-LDL (IC50: 1.7 +/- 0.7 versus 7.7 +/- 1.0 micrograms protein/ml). No significant differences among the different methods of iodination of LDL were found. The findings show that 111In-labeled lipoproteins might be a better ligand for lipoprotein-receptor binding studies as compared to radioiodinated lipoprotein products.

Adult↗
PubMed1990

myo-inositol oxygenase from rat kidneys. Substrate-dependent oligomerization.

myo-Inositol from rat kidneys, an oligomeric protein with apparent molecular mass of about 270 kDa can be dissociated under mild conditions to structured 16.8-kDa monomers. This dissociation can be reversed at high protein concentrations at room temperature. The corresponding apparent dimerization constant K2app = 1.38 x 10(5) M-1, the corresponding rate constant k2 = 350 s-1.M-1, and the apparent constant for the association of dimers, K4app = 2.7 x 10(6) M-1. Reassociation is significantly enhanced in the presence of the substrate and iron(II) (K2app = 9.8 x 10(5) M-1; K4app = 3.75 x 10(6) M-1, k2 = 1750 s-1.M-1, at 20 mM myo-inositol and 0.5 mM FeSO4). Under these conditions almost 100% of the original enzymatic activity was reconstituted. Monomers, with or without bound ligands, lack catalytic activity, whereas the dimer is likely to be the elementary active enzyme-building unit. The effects of myo-inositol on the dimerization lead to the conclusion that this step is both mediated and facilitated by the substrate.

Animals↗
PubMed1989

Purification and identification of the lipoprotein-binding proteins from human blood platelet membrane.

As reported previously, homologous plasma lipoproteins specifically bind to the plasma membrane of human blood platelets. The two major lipoprotein-binding membrane glycoproteins were purified to apparent homogeneity and identified by their mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, both in the nonreduced and reduced state, by specific antibodies against glycoproteins IIb (GPIIb) and IIIa (GPIIIa), respectively, including the alloantibody anti-PlA1 and monoclonal antibodies. Furthermore, lipoprotein binding to intact platelets is also inhibited in a dose-dependent fashion by preincubation of the platelets with antibodies against these glycoproteins. From these experiments it can be concluded that lipoproteins bind to both components of the glycoprotein IIb-IIIa complex in isolated membranes and intact platelets. High density lipoprotein and low density lipoprotein bind to GPIIIa blotted to nitrocellulose in a way that binding of one species interferes with the binding of the other. Addition of fibrinogen significantly inhibits this binding. The specific binding of fibrinogen to GPIIIa is strongly inhibited in the presence of either of the two lipoproteins. LDL and HDL are specifically bound by isolated GPIIb, too. In our blotting experiments fibrinogen shows no binding to this membrane glycoprotein. On the other hand, fibrinogen significantly interferes with the interaction between GPIIb and the lipoproteins.

Antibodies, Monoclonal↗
PubMed1989

The globular domains of type VI collagen are related to the collagen-binding domains of cartilage matrix protein and von Willebrand factor.

Type VI collagen is a transformation-sensitive glycoprotein of the extracellular matrix of fibroblasts. We have isolated and sequenced several overlapping cDNA clones (4153 bp) which encode the entire alpha 2 subunit of chicken type VI collagen. The deduced amino acid sequence predicts that the alpha 2(VI) polypeptide consists of 1015 amino acid residues that are arranged in four domains: a hydrophobic signal peptide of 20 residues, an amino-terminal globular domain of 228 residues, a collagenous segment of 335 residues and a carboxy-terminal globular domain of 432 residues. The collagenous domain contains seven Arg-Gly-Asp tripeptide units, some of which are likely to be used as cell-binding sites. The globular domains contain three homologous repeats with an average length of 180 amino acid residues. These repeats show a striking similarity to the collagen-binding motifs found in von Willebrand factor and cartilage matrix protein. We therefore speculate that the globular domains of the alpha 2(VI) polypeptide may interact with collagenous structures.

Amino Acid Sequence↗
PubMed1988

Ophthalmic observations in lecithin cholesterol acyltransferase deficiency.

Lecithin cholesterol acyltransferase is an enzyme that esterifies free cholesterol. A complete deficiency of this enzyme results in a diffusely cloudy cornea. This deficiency is thought to be transmitted as an autosomal recessive trait. We studied a family in which four members were homozygote recessive. In the homozygote recessive condition, a central corneal haze caused by deposition of numerous minute gray dots was consistently present. In the heterozygote condition, arcuslike changes were present in some of the patients studied. We found the corneal change in the recessive state to be sensitive and specific as a marker of this condition. Heterozygotes appear to have a higher incidence of arcuslike corneal changes.

Adolescent↗
PubMed1988

Fluorometric continuous kinetic assay of alpha-chymotrypsin using new protease substrates possessing long-wave excitation and emission maxima.

A direct and continuous kinetic method for the fluorometric determination of alpha-chymotrypsin and trypsin is described, and 2-aminoacridone (2-AA) is introduced as a promising new fluorophore in analytical biochemistry. N-Succinyl- and N-glutaryl-phenylalanine as well as N-benzoylarginine were coupled to 2-AA via a peptide bond and the resulting fluorogenic substrates are shown to be cleaved by the two enzymes. Since the substrate and product of hydrolysis have quite different spectral properties, the increase in the long-wave fluorescence of 2-AA (measured at 570 nm under 450-nm excitation) is a parameter for the enzyme activity. Chymotrypsin (0.5 microgram/ml) and trypsin (0.1 microgram/ml) were detectable in a 3-min assay. The major advantages of the new substrates over existing ones are the analytical wavelengths which are distinctly outside the background fluorescence of most biological matter and the somewhat faster reaction rates which can reduce the time of analysis.

Aminoacridines↗
PubMed1987

Investigation of lipid peroxidation in human low density lipoprotein.

Human plasma low density lipoprotein (LDL) exposed to oxygen saturated buffer becomes depleted of alpha-tocopherol within 3 to 6 hours. Thereafter, lipid peroxidation commences as evidenced by the loss of 18:2 (67 nmol/mg LDL) and 20:4 (12 nmol/mg LDL) and the concomitant formation of 4-hydroxynonenal (0.28 nmol/mg LDL) and fluorescent compounds. The major fluorophor in apo B of oxidized LDL has an excitation maximum at 355 nm and an emission maximum at 430 nm. A fluorophor with the same spectral properties is produced in apo B, if LDL is incubated with 4-hydroxynonenal, whereas malonaldehyde gives a fluorophor with excitation and emission maxima at 400/470 nm. Three-dimensional fluorescence spectroscopy proved to be an useful tool in analysing the complex fluorescence of apo B.

Aldehydes↗
PubMed1987

Autoxidation of human low density lipoprotein: loss of polyunsaturated fatty acids and vitamin E and generation of aldehydes.

The alteration of structural and biological properties of human plasma low density lipoprotein (LDL) exposed to oxidative conditions is in part ascribed to lipid peroxidation. The objective of this investigation was to measure quantitatively several parameters in oxidizing LDL indicative for lipid peroxidation. Exposure of freshly prepared EDTA-free LDL to an oxygen-saturated buffer led to a complete depletion of alpha- and gamma-tocopherol within 6 hr, thereafter lipid peroxidation commenced as indicated by the kinetics of the loss of linoleic (18:2) and arachidonic (20:4) acids, the formation of aldehydic lipid peroxidation products and fluorescent apoB. Within 24 hr of oxidation, on average 79 nmol of 18:2 (initial 345) and 12.8 nmol of 20.4 (initial 25.6) were oxidized per mg of LDL and the sample contained in total 7.1 nmol of aldehydes with the following molar distribution: 36.6% malonaldehyde, 25% hexanal, 8.9% propanal, 8.2% 4-hydroxynonenal, 7.6% butanal, 4.1% 2.4-heptadienal, 3.4% pentanal, 3.4% 4-hydroxyhexenal, and 2.5% 4-hydroxyoctenal. Malonaldehyde was predominantly (93%) in the aqueous phase, whereas the other aldehydes remained mostly (34-98%) within the LDL particle, where the total aldehyde concentration was in the range of 12 mM. Oxidized LDL exhibited a 1.6-fold enhanced electrophoretic mobility. Similarily, native LDL incubated for 5 hr with aldehydes showed increased electrophoretic mobility. At equal concentrations (5 mM) 4-hydroxynonenal was most effective, followed by 2,4-heptadienal, hexanal, and malonaldehyde. This study reports for the first time the rate and extent of the change of LDL constituents occurring during lipid peroxidation.

Aldehydes↗
PubMed1986

Possible involvement of the lipid-peroxidation product 4-hydroxynonenal in the formation of fluorescent chromolipids.

The effects of the lipid-peroxidation product 4-hydroxynonenal on the formation of fluorescent chromolipids from microsomes, mitochondria and phospholipids were studied. Incubation of freshly prepared rat liver microsomes or mitochondria with 4-hydroxynonenal results in a slow formation of a fluorophore with an excitation maximum at 360 nm and an emission maximum at 430 nm. The rate and extent of the development of the 430 nm fluorescence can be significantly enhanced by ADP-iron (Fe3+). With microsomes, yet not with mitochondria. NADPH has a catalytic effect similar to that of ADP-iron. Fluorescent chromolipids with maximum excitation and emission at 360/430 nm are also formed during the NADPH-linked ADP-iron-stimulated lipid peroxidation. Phosphatidylethanolamine and phosphatidylserine react with 4-hydroxynonenal revealing a fluorophore with the same spectral characteristics as that obtained in the microsomal and mitochondrial system. The findings suggest that the fluorescent chromolipids formed by lipid peroxidation are not derived from malonaldehyde, but are formed from 4-hydroxynonenal or similar reactive aldehydes via a NADPH and/or ADP-iron-catalysed reaction with phosphatidylethanolamine and phosphatidylserine contained in the membrane.

Adenosine Diphosphate↗