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Biomedical subjects

K Beyer

Publications and source records attributed to K Beyer.

83 records · Page 5Linked to original sources

ADP/ATP carrier protein from beef heart mitochondria has high amounts of tightly bound cardiolipin, as revealed by 31P nuclear magnetic resonance.

An unusual binding of cardiolipin to the ADP/ATP carrier has been found, which is distinguished by the relatively large amount and by the tightness of binding. High-resolution 31P NMR studies on the detergent-solubilized ADP/ATP carrier from beef heart mitochondria revealed narrow signals from phosphatidylcholine and phosphatidylethanolamine and a broadened signal of 30-40-Hz line width, suggestive of cardiolipin. Line broadening of this magnitude is to be expected when tumbling of the whole protein-detergent micelle is the only source of phosphorus spin-spin relaxation. Thus a strong immobilization of the protein-bound cardiolipin is inferred. By sucrose density gradient centrifugation phosphatidylcholine and phosphatidylethanolamine were removed, while approximately six +/- one molecules of cardiolipin remained tightly bound in the dimeric protein molecule. The cardiolipin binding was stable against treatment with sodium dodecyl sulfate although release of the inhibitor carboxyatractyloside revealed at least partial protein denaturation. Ca2+ ions did not readily interact either with the bound cardiolipin. Complete detachment of the bound phospholipid was achieved by a short heat pulse in the presence of sodium dodecyl sulfate. Denaturation of the carrier protein by guanidinium chloride or NaClO4 also led to release of the bound phospholipid. Thus different stages of protein denaturation must be envisaged.

Animals↗

[Normal lymph node cross sections in different anatomic regions and their significance for computed tomographic diagnosis].

In 102 patients, lymph node cross section measurements in different anatomic regions were taken from standard bipedal lymphograms without evidence of lymphatic disease. Corrections were made for geometric magnification due to anatomic changes in lymph node: film distances. In each region, the arithmetic mean, standard deviation, median value, Gaussian distribution, and 95% confidence limits were calculated. In 51 male patients (age range: 15 to 78 years) and 51 female patients (age range: 20 to 76 years), the following upper limits of normal lymph node cross section measurements were found: Retrocrural nodes - 7 mm, lumbar nodes - 10 mm, iliac lymph nodes - 12 mm, inguinal lymph nodes - up to 18 mm. The values obtained showed no statistically significant differences between male and female patients and between the right and left sides of the body. The problem of size as the only criterion of malignancy in computed tomography as well as in sonography of the retroperitoneal lymph nodes is discussed.

Adult↗

Biochemical expression of mosaicism in female mice heterozygous for the jimpy gene.

Expression of the jimpy gene in heterozygous females was analyzed by measuring galactolipid synthesis in brain during early myelination. Sulfatide labeling in brains of heterozygous females at 13-14 days is decreased to 40-80% that of female littermates who do not carry the jimpy gene. The activities of ceramide galactosyl transferase and cerebroside sulfotransferase and levels of myelin basic protein were similarly depressed. Since the jimpy gene was maintained with the Tabby gene in these studies, the effect of the Tabby gene on these parameters was examined and found to have no effect. These biochemical findings indicate that myelination is retarded in the brains of heterozygous jimpy females during the second week of development. However, at 18 days and thereafter, sulfatide labeling is less reduced, suggesting that the oligodendrocytes in brain attempt to compensate, in agreement with morphologic studies which show that myelin is decreased in brain during early development, but appears normal in adult animals.

Animals↗

Binding of spin-labeled carboxyatractylate to mitochondrial adenosine 5'-diphosphate/adenosine 5'-triphosphate carrier as studied by electron spin resonance.

The spin-label 2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid was attached to the inhibitor carboxyatractylate of the mitochondrial ADP/ATP carrier. Being closely linked to the inhibitor, the spin-label should reflect the mobility of the carboxyatractylate. When bound to the carrier in mitochondria, spin-labeled carboxyatractylate reveals a most unusual hyperfine splitting of 72 G. A second spectral component with a hyperfine splitting of 62 G is also mainly due to carrier-bound inhibitor. A similar spectrum with somewhat reduced hyperfine splitting was observed with the detergent-solubilized protein, whereas reincorporation into phospholipid membranes yielded almost the same spectra as in mitochondria. The carrier-bound spin-label is concluded to be highly immobilized. The less immobilized spectral component is discussed in terms of strongly anisotropic label motion. In addition, the unusual splitting is interpreted to indicate the highly polar environment of the nitroxide. The interpretations are supported by the temperature dependence, which indicates a reversible progressive spin-label mobilization up to 50 degrees C. Membrane-impermeable reducing agents showed that the spin-label is easily accessible from the aqueous phase.

Animals↗

Reincorporation of adenosine 5'-diphosphate/adenosine 5'-triphosphate carrier into phospholipid membranes. Phospholipid-protein interaction as studied by phosphorus-31 nuclear magnetic resonance and electron microscopy.

Combined phosphorus-31 nuclear magnetic resonance (31P NMR) and electron microscopic studies were performed on the ADP/ATP carrier protein from beef heart mitochondria. The protein was incorporated into phospholipids by addition of Triton-protein micelles to a lipid suspension or to the dry lipid. All of the phospholipid (egg phosphatidylcholine or mixtures of egg phosphatidylcholine and egg phosphatidylethanolamine) that contributed to the observed 31P NMR signal under these conditions appeared to be in a bilayer configuration. Freeze-fracturing and negative-staining electron microscopy showed unilamellar vesicles and multilayers. An isotropic signal could be attributed to vesicle rotation, judging from its sensitivity to increasing viscosity. The presence of small vesicles was also noticeable in the 31P NMR spectra of planar oriented membranes. In the presence of phosphatidylethanolamine, aggregation of protein particles was observed. Gel chromatography of the protein-Triton-phospholipid mixture revealed that, before Triton removal, large amounts of protein are associated with multibilayers. Separation of loaded and unloaded membranes by centrifugation in D2O showed that, upon stepwise addition, protein incorporates preferentially into unloaded liposomes. From these findings a mechanism of protein reincorporation was deduced.

Animals↗

A 31P- and 2H-NMR study on lecithins in liquid crystalline polyoxyethylene detergents.

Phosphatidylcholines were incorporated into hexagonal liquid crystalline mixtures of the non-ionic detergents Triton X-100 and octaethyleneglycoldodecylether with D2O. It is shown by nuclear magnetic resonance (NMR) that the phospholipids adopt the hexagonal liquid crystalline structure of the detergent host lattice. The anisotropic motion of the phospholipid headgroups seems to be unaffected, whereas the acyl chains are disordered. Increasing phospholipid concentration leads to separation of a lamellar phase. The lamellar structure is also preferred at elevated temperatures. Phosphatidylcholines with saturated acyl chains undergo a transition from the hexagonal liquid crystalline to an ordered lamellar state. The shape of the 31P-NMR signals suggests that pure gel phase phospholipid separates out. The headgroup region of this gel phase phospholipid becomes immobilized after a few weeks of storage below the transition temperature as judged from 31P-NMR. At the same time 2H-NMR exhibits a new signal from D2O undergoing slow isotropic motion. This behavior bears resemblance to the formation of a coagel in fatty acid-water systems.

Egg Yolk↗

Interaction of an amine oxide detergent with lecithin vesicles as studied by nuclear magnetic resonance.

The interaction of an amine oxide detergent with single bilayer lecithin vesicles was investigated with proton and phosphorus magnetic resonance. The addition of the detergent micelles to vesicles suspensions leads to rapid detergent incorporation into the vesicle bilayer, resulting in a heterogenous vesicle population. Initially, some vesicles take up the equivalent of one detergent micelle, whereas others contain no detergent. Subsequently, the detergent is distributed between the vesicles by vesicle-vesicle collisions. This can be followed by the change in the Pr3+-shifted spectral positions of the detergent and lecithin head groups with time. From the intensity of the head-group signals, it can be concluded that after about 20 h the detergent is almost equally distributed between the outer and inner vesicle membrane monolayers. Vesicles obtained by cosonication of the detergent and lecithin take up metal ions. This ion permeability depends on the vesicle concentration and can be attributed to vesicle-vesicle or vesicle-mixed micelle collisions. Egg lecithin vesicles are stable against the detergent up to molar ratios of detergent to lecithin of 0.2--0.3. At larger ratios mixed micells and multibilayers are formed. Measurements of proton spin-lattice relaxation times confirmed that the internal architecture of the vesicle bilayer is almost unaffected by the incorporated detergent.

Detergents↗

Studies on the tolerance of silk, nylon, dacron and collagen suture material in the cornea of the rabbit.

With rabbit corneas we did comparative biomicroscopical and histological investigations on material tolerance of the following threads at present used in microsurgery: Barraquer silk (virgin silk), nylon 30 mum (Tübinger nylon), dacron (polyester) 40 mum and plain collagen 7-0. The sutures were implanted into the cornea intra-lamellarly, completely under the microscope. The examinations showed only very little tissue reaction in all these materials.

Animals↗