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Biomedical subjects

M Müller

Publications and source records attributed to M Müller.

At least 1,189 records · Page 66Linked to original sources
PubMed1988

Secondary structure determination for the Antennapedia homeodomain by nuclear magnetic resonance and evidence for a helix-turn-helix motif.

The homeodomain encoded by the Antennapedia (Antp) gene of Drosophila was studied in aqueous solution by nuclear magnetic resonance (NMR). Sequence-specific resonance assignments have been obtained for the complete polypeptide chain of 68 amino acid residues. The secondary structure determined from nuclear Overhauser effects (NOE) and information about slowly exchanging amide protons includes three helical segments consisting of the residues 10-21, 28-38 and 42-52, respectively. Combination of the presently available NMR data with computer modeling provided preliminary evidence for the presence of a helix-turn-helix motif in the homeodomain. Near the turn, this supersecondary structure appears to be very similar to the DNA binding site in the 434 and P22 c2 repressors, but both helices in the homeodomain include 2-3 additional residues when compared with these prokaryotic DNA-binding proteins.

Amino Acid Sequence↗
PubMed1988

Preparation of a one-subunit cytochrome oxidase from Paracoccus denitrificans: spectral analysis and enzymatic activity.

Cytochrome c oxidase was isolated from Paracoccus denitrificans as a two-subunit enzyme. Chymotrypsin-catalyzed proteolysis reduced the molecular weight of each subunit by about 8000. The spectral properties of this preparation, as well as its Km for cytochrome c(1.7 muM), remained unchanged with respect to the native enzyme. Vmax was reduced by about 55% when assayed in Triton X-100 or in Triton X-100 supplemented with asolectin. Following further proteolysis by Staphylococcus aureus V8 protease, subunit I remained unchanged as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whereas subunit II was split into small peptides. These were removed by ion-exchange high-performance liquid chromatography. The one-subunit enzyme had an apparent molecular weight of 43,000. The reduction of molecular weight was also confirmed by the diminution of the ultraviolet/Soret absorption ratio. This value was 1.8-2.1 for the native enzyme and 1.3-1.5 for the one-subunit enzyme. The spectral properties (including the spectrum CO reduced minus reduced) were not modified by the proteolytic treatment, indicating that cytochromes a and a3 were present in equal amounts. The lack of spectral alteration and the known close association of the copper B atom with cytochrome a3 suggest that copper B is also contained within the one-subunit enzyme. The Km of the one-subunit oxidase was similar to that of the two-subunit enzyme; Vmax was decreased by about 50%. The activity of the one-subunit oxidase had a salt-dependent maximum at 30 mM KCl, almost identical with that of the undigested enzyme, and was inhibited by micromolar concentrations of KCN.

Affinity Labels↗
PubMed1988

Subunit III of cytochrome c oxidase is expressed in Paracoccus denitrificans.

Polyclonal antibodies have been obtained against a synthetic dodecapeptide identical to the aminoacid sequence 120-131 DSPIKDGVWPPE (inferred from its DNA sequence) of Paracoccus denitrificans cytochrome c oxidase subunit III. The antibodies had a titer higher than 1:10000 when tested against the antigen. These antibodies have been used to produce immunological evidence that, despite the fact that subunit III is not isolated with cytochrome c oxidase, it exists in Paracoccus denitrificans lysates. The antibodies did not show reactivity with bovine heart cytochrome c oxidase either by ELISA or immunoblotting. It was also shown that the antibodies react with a single polypeptide present in Paracoccus denitrificans cell lysates, having an apparent molecular weight close to that of subunit III of bovine heart oxidase.

Animals↗
PubMed1988

The effect of non-esterified fatty acids on the proton-pumping cytochrome c oxidase reconstituted into liposomes.

Bovine heart cytochrome c oxidase was reconstituted in phospholipid vesicles, and the effect of different non-esterified fatty acids (NEFA) was studied on its proton pump and on the proton permeability of the vesicles. Neither parameter appeared to be affected by concentrations of NEFA known to uncouple oxidative phosphorylation (10 microM). Also the permeability for K+ was not affected by them. The fatty acids caused an increase in the rate of electron transfer in the absence, but not in the presence, of uncoupler and/or valinomycin [diminution of the respiratory-control index (RCI)]. The RCI of 8.7-7.5 was decreased to about 4.5 in the presence of 0.27-10 microM-NEFA. Oleic acid was not effective at the above concentrations. Subunit III-depleted enzyme preparations gave vesicles with an RCI of about 5.5, which was decreased to 4.5 in the presence of NEFA. With both native and subunit III-depleted oxidase the RCI was never decreased to the value of 1 by NEFA, as happens with classical protonophores.

Animals↗
PubMed1988

[Surgery of colon and rectal cancer].

The modern surgery of the colorectal carcinoma is becoming more and more standardized which is reflected by the drastic reduction of postoperative lethality. Since the curing rates have only been marginally improved in the last 20 years the new trend is going in the direction of more radical techniques. Adjuvant therapy methods such as radio-, chemo- and immunotherapy have not yet fulfilled their expectancies. The modern standard and problematic in the colorectal carcinoma surgery is discussed using the results of 257 operated carcinomas in the time from January 1st 1984 to April 28th 1987.

Adenocarcinoma↗
PubMed1988

[Crohn's disease-- 10 years' experience at a city hospital].

Between 1977 and 1986, 92 patients with Crohn's disease (53 females and 39 males, average age 32 years at diagnosis) were admitted for a total of 158 hospital stays at the St. Claraspital, Basel. In 71 cases the diagnosis was based on one or more of radiologic and endoscopic evaluation and/or operative findings. In 4 cases diagnosis was based solely on clinical findings and in the remaining 17 the original diagnostic workup could not be evaluated retrospectively. 65% of cases were histologically proven, while the rest of the patients were either not biopsied or biopsy yielded inconclusive results. Crohn's disease was located simultaneously in the small and large bowels in 41 patients, in the small bowel only in 23 (25%) and in the large bowel only in 20 (22%). The remaining localizations were either proximal GI tract or anal region only. Surgery was performed a total of 65 times in 44 of the 92 patients (48%), the most frequent procedures being ileocecal resection and surgical treatment of perianal fistulas and abscesses. Accordingly, the main indications for surgery were perianal complications (37%) and stenotic bowel lesions (29%). The resection rate was higher in ileal (35%) than in colorectal (25%) involvement. The 3 rectal amputations eventually necessary were all in patients with anorectal involvement.

Adult↗
PubMed1988

Identification of a myometrial oxytocin-receptor protein.

The specific binding of [3H]oxytoxin to uterine membrane preparations derived from different species at late pregnancy was examined. The highest receptor density (bmax value) was found in membranes derived from the myometria of guinea pigs between day 60 post-conception (bmax = 3.6 +/- 0.1 pmol/mg) and day 65 (bmax = 4.4 +/- 0.1 pmol/mg). The similarity of Kd values for oxytocin binding (Kd = 2.6 +/- 0.2 nM) and for vasopressin binding (Kd = 2.1 +/- 0.4 nM) to the same membranes derived from a guinea pig myometrium indicate a homogeneous population of high-affinity binding sites which do not discriminate between these two hormones. Competitive binding experiments with specific oxytocin agonists containing either sarcosine or N-methylalanine in the place of Pro7 demonstrated that these myometrial receptors have the pharmacological properties of oxytocin receptors. The analogue of 1-deamino-[8-lysine]vasopressin containing a photoreactive azidophenylamidino group at the sidechain of Lys8 retained roughly the same receptor affinity as oxytocin. In photoaffinity labelling experiments with the tritium-labelled analogue a membrane protein from guinea pig myometrium with an apparent relative molecular mass Mr of 78,000 +/- 5000 (n = 13) was preferentially labelled. The labelling of this protein was completely suppressed by a 100-fold molar excess of either oxytocin, or [Sar7]oxytocin or [Thr4, Sar7]oxytocin, but not by other peptide hormones. These results provide evidence that the labelled 78,000-Mr protein is a myometrial oxytocin-receptor protein.

Affinity Labels↗
PubMed1988

Processing by inverted plasma membrane vesicles of in vitro synthesized major lipoprotein from Escherichia coli.

Synthesis, lipid modification and proteolytic processing of the major lipoprotein from Escherichia coli is shown to occur in a homologous in vitro transcription-translation system containing inverted plasma membrane vesicles. The primary translation product (cross-reacting with anti-lipoprotein antiserum) is a precursor which is converted into a lower molecular mass species of the size of mature lipoprotein by the addition of inverted membrane vesicles from E. coli. Conversion is prevented by globomycin, a specific inhibitor of the unique lipoprotein-signal peptidase II, which is active only on lipid-containing precursors. The inverted plasma membrane vesicles used here must therefore contain active lipid-modifying enzymes and signal peptidase II.

Cell Membrane↗
PubMed1988

Origin of cochlea efferents in some gerbil species. A comparative anatomical study with fluorescent tracers.

The distribution and the projection patterns of olivo-cochlear neurons (OCN) was studied in three species of gerbils (Meriones unguiculatus, Meriones tristrami, Pachyuromys duprasi) using fluorescent tracers (Fast blue, Diamidino yellow) in double labeling experiments. In all gerbil species up to 1600 retrogradely labeled neurons were found in the superior olivary complex. Small OCN were located inside or at the margins of the lateral superior olive. A second set of larger OCN was seen bilaterally in the periolivary region of the superior olivary complex forming an independent cell population within the trapezoid body. The exact locations of small and large OCN varied among the particular species. Quantitative results suggested that a gradual loss of large OCN might be a general feature of species with highly specialized frequency selectivity.

Amidines↗
PubMed1988

A small population of retinal ganglion cells projecting to the retina of the other eye. An experimental study in the rat and the rabbit.

We traced the retino-retinal projection with Rhodamine B isothiocyanate (RITC), Rhodamin labelled latex microspheres (RLM), horseradish peroxidase (HRP) and choleratoxin conjugated horseradish peroxidase (BHRP). The number and distribution of ganglion cells projecting to the contralateral eye were recorded. Newborn and young rats have up to about 130 ganglion cells projecting to the other retina; this confirms previous findings. We extended these findings in two ways. First, we describe a similar projection in rabbits consisting of fewer cells; second, we describe the persistence of a small component of this projection into adulthood. In addition we show with RITC and Nuclear Yellow double tracing that some of the retino-retinal ganglion cells have an axon collateral which projects to the superior colliculus. We performed control experiments in order to exclude spillover of tracer which might produce false positive labelling.

Animals↗
PubMed1988

Ultrastructural demonstration of lectin binding sites in the Golgi apparatus of rat epiphyseal chondrocytes.

Binding sites for wheat germ agglutinin (WGA), Dolichos biflorus agglutinin (DBA), Ricinus communis I agglutinin (RCA I) and Limax flavus agglutinin (LFA) have been ultrastructurally detected in rat epiphyseal chondrocytes by a post-embedding cytochemical technique using colloidal gold as marker. The four lectins labelled exclusively the Golgi apparatus of chondrocytes embedded in Lowicryl K4M resin by two different methods. WGA binding sites were localized in medial and trans cisternae as well as in immature secretory vesicles, whereas those for DBA were seen concentrated in cis and medial cisternae. Labelling with both RCA I and LFA lectins was distributed throughout all the cisternae of the Golgi stack, and the latter also in vesicles and tubules at the trans face. Neuraminidase pretreatment of the sections abolished LFA staining, decreased reaction with WGA and increased that with RCA I, while it did not affect DBA staining. After chondroitinase ABC treatment only the RCA I reaction was modified, revealing new binding sites in the trans Golgi face, secretory granules and extracellular matrix. These results indicate that the distribution of subcompartments in the Golgi apparatus of chondrocytes is different from that in cells secreting glycoproteins as major products.

Animals↗
PubMed1988

Transport systems for amphipathic compounds in normal and neoplastic hepatocytes.

Photoaffinity labeling of plasma membrane subfractions from liver and of intact liver tissue with a photolabile bile salt derivative, the sodium salt of (7,7-azo-3 alpha,12 alpha-dihydroxy-5 beta-cholan-24-oyl)-2-aminoethanesulfonic acid, revealed that the hepatobiliary transport of bile salts is accomplished by transport systems different for sinusoidal uptake and canalicular secretion. Polypeptides with apparent Mr values 54,000 and 48,000 interact with bile salts at sinusoidal membrane, whereas a polypeptide with an apparent Mr of 100,000 is involved in bile salt secretion through the canalicular membrane. Photoaffinity labeling with photolabile derivatives of uncharged and cationic compounds provided evidence that the sinusoidal membrane polypeptides exhibit a broad binding specificity. Photoaffinity labeling studies and kinetic studies suggest that hepatic uptake of different amphipathic anions, uncharged compounds and even of cations is mediated by the sinusoidal transport systems which are involved in the uptake of bile salts. Relatively little is known about the specificity of the canalicular bile salt transport system. The fluorescent bile salt derivative, the sodium salt of (N-[7-(4-nitrobenzo-2-oxa-1,3-diazol)]-3 beta-amino-7 alpha, 12 alpha-dihydroxy-5 beta-cholan-24-oyl)-2-aminoethanesulfonic acid, is readily taken up into the hepatocytes of all acinar zones and may be used for the evaluation of the functional state of bile salt transport by fluorescence microscopy. Fluorescent microscopic studies with the fluorescent bile salt derivative showed that ascites hepatoma AS 30D cells do not have the ability to take up bile salts and demonstrated the absence of hepatobiliary bile salt transport in the solid Morris hepatoma 7777. Photoaffinity labeling studies revealed that in both tumor cell models, in hepatoma AS 30D and in Morris hepatoma 7777, the plasma membranes were devoid of the polypeptides having affinities to bile salts and amphipathic cations. A slight labeling of bile salt binding membrane polypeptides in plasma membranes from Morris hepatomas 9618A and TC 5123 opens the possibility to study transport in neoplastic hepatocytes.

Animals↗
PubMed1988

Ornithine decarboxylase in the inner ear of the guinea pig.

L-Ornithine decarboxylase, the rate limiting enzyme of polyamine synthesis and a possible marker enzyme for tissue proliferation and maturation, has been found in the developing guinea pig cochlea using the unlabelled horseradish-peroxidase-antiperoxidase technique. Ornithine decarboxylase-like immunoreactive material was detected in the neurons of the Ganglion spirale and in their axonal and/or dendritic fibers. The location of the enzyme and the possible functional role of ornithine decarboxylase plays in the development and maturation of the auditory organ and of the hearing process are discussed.

Aging↗
PubMed1988

Inner ear structure and electrophysiological audiograms of the subterranean mole rat, Spalax ehrenbergi.

Subterranean mole rats of the Spalax ehrenbergi superspecies in Israel have a distinctly developed vocal repertoire, presumably compensating together with olfaction for their complete blindness, thus providing an efficient communication system underground. Here we describe the unique organization of the cochlea of Spalax among mammals. The cochlea is subdivided into different subsystems where in the apical subsystem the fluid space and the organ of Corti differ remarkably from that in the basal subsystem, a feature as yet unknown in other mammals. The audiograms based on cochlear microphonics and on evoked potential recordings from the midbrain and brainstem, reveal a hearing range from 0.1 kHz-10 kHz with a best sensitivity between 0.5 and 1 kHz.

Acoustic Stimulation↗