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Biomedical subjects

M Hogan

Publications and source records attributed to M Hogan.

At least 73 records · Page 4Linked to original sources
PubMed1988

Site-specific derivatives of wheat germ calmodulin. Interactions with troponin and sarcoplasmic reticulum.

Wheat germ calmodulin (CaM) was derivatized at its single cysteine (Cys27) with either the fluorescent reagent, N-(iodoacetylaminoethyl)-5-naphthylamine-1-sulfonic acid (I-EDANS) or the photoactivable cross-linker benzophenone-4-maleimide. Comparison of the native and derivatized wheat germ CaMs with native bovine testis CaM indicates that the concentrations of these proteins required for half-maximal stimulation of either erythrocyte membrane Ca2+-ATPase activity or cardiac sarcoplasmic reticulum phosphorylation are very similar. Affinity labeling of troponin subunits with 125I- and benzophenone-4-maleimide-labeled CaM demonstrates CaM binding to troponin I (TnI) and troponin T (TnT) in binary complexes, as well as to both subunits in the CaM.TnI.TnT ternary complex. This suggests that both subunits are within 10 A of Cys27 of calmodulin. Affinity labeling of cardiac sarcoplasmic reticulum vesicles with 125I- and benzophenone-4-maleimide-labeled CaM exhibits a Ca2+- and Mg2+-dependent labeling of phospholamban, as shown previously with bovine calmodulin (Louis, C.F., and Jarvis, B. (1982) J. Biol. Chem. 257, 15187-15191). Thus, it appears that Ca2+-binding site I of calmodulin is at or near binding sites of calmodulin for TnI, TnT, and phospholamban. Analysis of the time-resolved fluorescence decay curves of I-EDANS-labeled calmodulin indicates a major component with a lifetime of 11.9 ns (+Ca2+), which accounts for 81% of the total fluorescence. The lifetime decreases slightly to 11.3 ns in the absence of Ca2+. Fluorescence anisotropy experiments indicate that I-EDANS-labeled CaM binds TnI with Kd = 6 x 10(-8) M in the presence of Ca2+. This study suggests that these single-site derivatives will be useful for characterizing a variety of calmodulin-receptor interactions because they lack ambiguities inherent in less specific labeling methods.

Animals↗
PubMed1987

A nuclear specific glycoprotein representative of a unique pattern of glycosylation.

Whole rat liver nuclei were reacted with UDP-[14C]galactose in the presence of bovine beta(1----4) galactosyltransferase. The reaction mixture was electrophoresed on a reducing sodium dodecyl sulfate-polyacrylamide gel. Autoradiograms of the gel demonstrated a major labeled broad band migrating with an apparent molecular weight of 65,000-66,000. A number of other less prominently labeled bands were also present. The labeled 65,000-66,000 band when cut from the gel and subjected to alkaline reduction while in the gel matrix exclusively yielded a 14C-labeled disaccharide that co-migrated with a [14C]Gal-GlcNAcol standard in descending paper chromatography. Treatment of this disaccharide with beta-galactosidase (beta-D-galactoside galactohydrolase; EC 3.2.1.23) from Aspergillus niger removed all the [14C]galactose label. Treatment of the labeled 65,000-66,000 polypeptide with Endoglycosidase F, however, did not remove the [14C]galactose label. Western transfer blots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels performed with horseradish peroxidase-labeled succinyl wheat germ agglutinin, a lectin specific for GlcNAc, on unlabeled nuclei revealed a dominant band at 63,000-64,000. Subjecting 14C-labeled nuclei to this procedure resulted in a shift of the major horseradish peroxidase-labeled succinyl wheat germ agglutinin band to 65,000-66,000. The shifted band was coincident with the [14C]galactose band as visualized on an autoradiogram. A survey of other rat tissue nuclei revealed the same spectrum of [14C]galactose acceptor proteins with a dominant 65,000-66,000 galactose-labeled band.

Animals↗
PubMed1986

Properties of the 23,000-Da phosphoproteins in cardiac sarcolemma and sarcoplasmic reticulum.

The calmodulin- and cAMP-dependent protein kinase-mediated phosphorylations of isolated sarcolemma and sarcoplasmic reticulum vesicles have been compared. Similarities in the calmodulin-mediated phosphorylation of the sarcolemma and sarcoplasmic reticulum 23,000-Da phosphoproteins included their Mg2+, Na+, Ca2+, and calmodulin sensitivities, as well as the size of their dissociated subunits. In contrast, a number of differences between these phosphoproteins were indicated in their sensitivity to detergents (Triton X-100 and sodium dodecyl sulfate) and calmodulin antagonists (R24571 and trifluoperazine). Furthermore, in contrast to the sarcoplasmic reticulum phosphoprotein, the sarcolemma phosphoprotein could not be affinity labeled with 125I-calmodulin. While these results indicate the probable chemical similarity of the sarcolemma and sarcoplasmic reticulum 23,000-Da phosphoproteins, they also indicate there are differences in the lipid/phosphoprotein interactions in these two membranes.

Alamethicin↗
PubMed1985

[Catheter occlusion during intraperitoneal infusion of insulin with implanted infusion devices].

Improvement in blood sugar control can be achieved, even in patients difficult to treat, using implantable insulin infusion devices for basal insulin infusion, together with one to two additional insulin injections. However, problems involving the insulin infusion route may arise, especially when insulin is infused intraperitoneally. Catheter adhesions and insulin precipitations can occur. Catheter blockages due to insulin aggregates can largely be avoided by the use of neutral sodium dihydrogen phosphate buffer. Insulin aggregations or precipitations are readily dissolved in vivo by alkaline buffer.

Adult↗
PubMed1985

An equilibrium between distorted and undistorted DNA in the adult chicken beta A-globin gene.

We have used single strand specific nucleases to map DNA distortion in the adult chicken beta A-globin gene. We have detected two structures of that kind and have mapped nuclease-cutting sites at one base resolution. One prominent site is centered at -190 relative to the RNA capping site and is positioned at the center of a stretch of contiguous C residues. The second site is near the first intron/exon junction (+620) and appears as a series of discrete 1-base-long enzyme-cutting sites. Based upon the pattern of nuclease cutting and the kinetics of nuclease cutting we conclude that the "poly(C)" stretch may assume a looped geometry in supertwisted DNA molecules which is similar to that proposed by Felsenfeld (Nickol, J. M., and Felsenfeld, G. (1983) Cell 35, 467-477). We show that S1 nuclease cuts within the intron occur mainly at the end points of polypurine segments and suggest that such end points may assume a distorted transitional geometry. We find that Neurospora crassa endonuclease cuts both the promotor and intron sites in linear DNA molecules but that in linear DNA the cutting process is limited by a first order conformation change of the DNA substrate. Based upon those kinetics we propose that in unstressed DNA, each of the two sites can convert between a distorted and undistorted geometry. In the enzyme assay buffer at 37 degrees C, the time constant for the equilibrium is nearly 10 h for the promotor site and 7 h for the intron.

Animals↗
PubMed1985

Cyanide intoxication among silver-reclaiming workers.

Thirty-six former workers in a silver-reclaiming facility who had been exposed over a long-term to excessive levels of cyanide were studied to determine acute and residual toxic reactions. The study involved physical examinations, laboratory studies, and a questionnaire to determine levels of exposure, symptoms during employment, and current symptoms. Questionnaire data showed that during the time of active employment there was a high prevalence of symptoms that are consistent with acute cyanide toxic reactions. A significant positive trend for prevalence of cyanide-related symptoms measured against levels of exposure was demonstrated, supporting a dose-response effect. Some symptoms occurring seven or more months after exposure had ceased also exhibited a dose-response trend. Mild abnormalities of vitamin B12, folate, and thyroid function were detected and suggest long-term cyanide effects.

Adult↗
PubMed1985

Lateral diffusion in nuclear membranes.

Chemical modification of rat liver nuclei with citraconic anhydride selectively removed outer nuclear membrane. This conclusion was based on (a) transmission electron microscopy, (b) lipid analysis, (c) lamin B as an inner membrane-associated marker, and (d) the demonstration of phospholipid lateral mobility on outer membrane-depleted nuclei as a criteria for inner membrane integrity. Addition of urea or N-ethylmaleimide resulted in the additional disruption of inner membrane. Fluorescence photobleaching was used to determine the long range (greater than 4 microns) lateral transport of lectin receptors and a phospholipid analog in both membranes. The diffusion coefficient for wheat germ agglutinin on whole nuclei was 3.9 X 10(-10) cm2/s whereas the diffusion coefficient for wheat germ agglutinin in outer membrane-depleted nuclei was less than or equal to 10(-12) cm2/s. Phospholipid mobilities were the same in whole and outer membrane-depleted nuclei (3.8 X 10(-9) cm2/s). The protein diffusion differences observed between whole and outer membrane-depleted nuclei may be interpreted in the context of two functionally different membrane systems that compose the double bilayer of the nucleus.

Animals↗
PubMed1985

The cultural perspective of therapeutic relationship--a viewpoint from Africa.

Therapeutic relationship has been considered an important ingredient of all psychotherapies. In communities in which no familiar conventions of such a relationship are available, the therapeutic encounter poses very different problems from those in the West, where such conventions freely prevail. This study has been carried out by five therapists representing three widely disparate cultures, but all working together in Tanzania. It brings together their perceptions of these problems and the strategies they employed to resolve them while working with African patients. In their view, in spite of great disparity between the world view behind Western psychotherapy and that of African communities, it is not impossible to forge a therapeutic relationship if empathic understanding and cultural sensitivity are added to the attitude of acceptance. After all, the therapist must attract and keep the patient before he can expect anything from him. The authors describe how this can be done with African patients.

Cross-Cultural Comparison↗
PubMed1984

Nuclear magnetic resonance imaging in gynecology.

Nuclear magnetic resonance proton imaging is a new imaging technique that holds promise for gynecologic diagnosis. Without the use of ionizing radiation, it provides images with excellent definition of the major pelvic organs. The extent and nature of disease are well demonstrated. In this report, an introduction to the principles of nuclear magnetic resonance imaging is given and several clinical examples that reveal the potential uses of nuclear magnetic resonance imaging in the female pelvis are shown.

Adult↗
PubMed1983

Triplet anisotropy decay measurements of DNA internal motion.

Triplet anisotropy decay techniques have been used to measure the internal flexibility and overall rotational motions of DNA over a time range of 15 ns to 200 microseconds. Nearly monodisperse DNA fragments with lengths varying from 65 to 600 base pairs were studied using the intercalating dye methylene blue as a triplet probe. The slow end-over-end tumbling of short DNA fragments (less than 165 base pairs) is as predicted for a rigid rod. As expected, a longer DNA fragment (600 base pairs) experiences slow segmental motions of its helix axis. At the earliest times, anisotropy decays more rapidly than expected for a rigid rod, suggesting that, when it is bound, methylene blue monitors fast internal motions of the helix. Since the rod-like end-over-end tumbling rules out fast bending motions (for short DNA fragments), the fast components of DNA anisotropy decay must be due to twisting motions of the helix, occurring with a time constant near 50 ns. The same techniques were used to measure the conformational flexibility of DNA in the nucleosome. It is concluded that, when the DNA helix is wrapped to form a nucleosome, it experiences substantial internal flexibility, occurring with a time constant near 30 ns. The amplitude and time-scale of this motion appear to be similar to that seen in the uncomplexed DNA helix.

Animals↗
PubMed1982

Molecular motion of DNA as measured by triplet anisotropy decay.

We have used triplet anisotropy decay techniques to measure the internal flexibility and overall rotational motion of DNA, covering a time range from 15 ns to 200 mus. Nearly monodisperse DNA fragments 65--600 base pairs long were studied by using the intercalating dye methylene blue as a triplet probe. We found that the slow end-over-end tumbling of short DNA fragments (less than or equal to 165 base pairs) is as predicted for a rigid rod. As expected, a longer DNA fragment (600 base pairs) experiences slow segmental motion of its helix axis. We found that, at the earliest times, anisotropy decays more rapidly than expected for a rigid rod, suggesting that, when bound, methylene blue monitors fast internal motion of the helix. Since the rod-like end-over-end tumbling of short fragments rules out fast bending motions, we conclude that the fast components of DNA anisotropy decay are due to twisting motion of the helix, occurring with a time constant near 50 ns.

DNA↗
PubMed1982

DNA motions in the nucleosome core particle.

We have used time-resolved triplet state anisotropy decay techniques to measure the conformational flexibility of DNA in the nucleosome. From these measurements we conclude that, in a nucleosome, the DNA helix experiences substantial internal flexibility, which occurs with a time constant near 30 nsec. We find that our data can be fit well by a modified version of the Barkley-Zimm model for DNA motion, allowing only DNA twisting motions and the overall tumbling of the nucleosome. That fit yields a calculated torsional rigidity equal to 1.8 X 10(-19) erg X cm, a value equal to that measured for uncomplexed DNA. We conclude from such similarity that large, fast twisting motions of the DNA helix persist, nearly unaltered, when DNA is wrapped to form a nucleosome.

Animals↗