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Biomedical subjects

C J Hsu

Publications and source records attributed to C J Hsu.

At least 55 records · Page 3Linked to original sources

Polyamines and autocrine control of tumor growth by prolactin in experimental breast cancer in culture.

We have previously observed that both polyamines and an autocrine mechanism are involved in the stimulation by ovine PRL (oPRL) of growth of the N-nitrosomethylurea-induced rat mammary tumor cultured in vitro in the soft agar clonogenic assay. These experiments were designed to test the interaction between these two pathways of oPRL action in this system. In agreement with previous findings in our laboratory, conditioned medium obtained from oPRL-treated (oPRL-CM), but not untreated, tumors consistently stimulated colony formation when added to N-nitrosomethylurea mammary tumors plated in soft agar under serum-free medium conditions. Administration of alpha-difluoromethyl-ornithine (DFMO), an irreversible inhibitor of polyamine biosynthesis, abolished the colony-stimulating effect of oPRL-CM. The inhibitory effect of DFMO was reversed in a dose-dependent fashion by the addition of spermidine, which entirely restored the growth-promoting action of oPRL-CM. In addition, the administration of increasing amounts of spermidine potentiated the colony-stimulating effect of suboptimal concentrations of oPRL-CM. In contrast, manipulation of the polyamine environment with DFMO and/or spermidine administration did not affect the number of colonies formed when conditioned medium from untreated tumors was added instead of oPRL-CM. We conclude that the polyamine pathways plays an essential role in the expression of autocrine control of tumor growth by oPRL.

Animals↗

The effect of relaxin on cyclic adenosine 3',5'-monophosphate concentrations in rat myometrial cells in culture.

Relaxin, a uterine relaxant secreted by the corpus luteum, was able to elevate cAMP concentrations in the presence of 1-methyl-3-isobutyl xanthine (MIX) (0.1 mM) or forskolin (0.4 microM) in a time- and dose-dependent manner in rat myometrial cells in culture but not in stromal cells. The optimal culture conditions for the cAMP response were determined to be an initial plating density of 1-1.5 X 10(6) cells/ml (3 ml/35-mm dish) and a 2-day culture period. In the presence of MIX, the time course of cAMP elevation in response to relaxin exhibited a lag phase of more than 5 min before cAMP concentrations rose significantly. However, in the presence of forskolin, relaxin elevated cAMP within 1 min. The concentration-response relationships were almost identical in the presence of MIX or forskolin. Isoproterenol was able to increase cAMP concentrations in myometrial cultures in both the absence and presence of MIX and to elevate cAMP levels rapidly within 1 min. These data suggest that cAMP could play some role in the initiation of uterine relaxation mediated by relaxin.

Animals↗

Carbonic anhydrase activity in the inner ear.

The histochemical and cytochemical localization of carbonic anhydrase activity in the inner ear of guinea pig was studied with Hansson's method and modified Yokota's method. All cells possessing the cell membrane infoldings are proved to have the enzymatic activity. The cochlear hair cells show strong enzymatic activity while the vestibular hair cells show no appreciable activity. The supporting cells of both cochlear and vestibular hair cells also show the enzymatic activity. The reaction products showing the enzymatic activity are observed in the cytoplasm, nucleus and cell membrane of the stained cells with both light microscope and electron microscope. The stria vascularis, vestibular dark cells and endolymphatic sac are proved to have the enzymatic activity. The cells, which are already proved to have Na+-K+-ATPase activity, show the enzymatic activity. The regulation of the fluid and ion in the inner ear is probably a complex mechanism involving not only various cells but also several enzymes such as carbonic anhydrase, Na+-K+-ATPase and adenylate cyclase.

Animals↗

Ultrasound-accelerated immunoassay, as exemplified by enzyme immunoassay of choriogonadotropin.

The rate-limiting step in many solid-phase immunoassays is associated with the slow kinetics of binding of macro-molecular antigen and conjugate to the immobilized phase. We demonstrate that the use of ultrasonic energy to enhance mass transport across liquid/solid interfaces can dramatically accelerate antigen binding to immobilized antibodies. We use an ultrasound-accelerated procedure with an enzyme-channelling test strip containing glucose oxidase and specific antibody to the alpha-subunit of human choriogonadotropin (HCG) co-immobilized onto a cellulose support. A horseradish peroxidase conjugate of monospecific antibody to the beta-subunit of HCG is used in the liquid phase to complete the immune "sandwich." Use of ultrasound to accelerate binding and of enzyme channelling to eliminate wash steps result in a simple two-incubation protocol by which 25 int. units of urinary HCG per liter can be detected visually in less than 20 min of assay time.

Chorionic Gonadotropin↗

Identification of functional domains of human erythrocyte spectrin.

Isolated human erythrocyte spectrin is a dimer of two unique polypeptide chains. The dimer (alpha beta) undergoes reversible salt- and temperature-dependent association to form (alpha beta)2 tetramers. Spectrin also binds with high affinity to a protein receptor on the cytoplasmic surface of erythrocyte membrane vesicles. By cleavage of spectrin at its cysteine residues with 2-nitro-5-thiocyanobenzoic acid, a 50,000-dalton peptide fragment has been isolated which inhibits the binding of spectrin to erythrocyte membrane vesicles. This peptide arises from a terminal region of the beta chain. An 80,000-dalton peptide generated by restricted trypsin digestion binds preferentially to dimeric spectrin. This peptide arises from a terminal portion of the alpha chain. Multiple peptides involved in noncovalent associations between the chains have also been identified. These associations indicate that the two subunits of spectrin are aligned parallel to one another and that the tetramer formation site and the high-affinity membrane binding site are in close proximity to one another.

Binding Sites↗

Substructure of human erythrocyte spectrin.

The human erythrocyte structural protein spectrin and its subunits I, II were isolated in the presence of Na-dodecyl-sulfate by gel filtration and preparative gel electrophoresis. After removal of the detergent, spectrin alpha-helical content is comparable to spectrin isolated without detergent. Subunits I and II formed single bands in isoelectric focusing (pI = 5.6) and in Ornstein-Davis disc gel electrophoresis systems, indicating the individual subunits are homogenous in nature. The molecular weights of the subunits I and II, determined by Ferguson plot, are 237,500 and 238,600, respectively, which is in good agreement with values obtained by the standard SDS gel relative mobility method. Limited tryptic digestion of spectrin and two-dimensional peptide maps of the individual subunits cleaved by S-cyanylation reaction showed dissimilar patterns, suggesting differences in primary structure between the two subunits.

Amino Acids↗

Neurogenic regulation of lipid metabolism in the rabbit. A mechanism for the cholesterol-lowering effect of acupuncture.

Plasma and aortic cholesterol levels are known to fall significantly after needling at the specific point which is innervated by a branch of the deep peroneal nerve. Different types of stimulation were applied to the specific nerve, using solely needling, injection of normal saline with needling and injection of a small dose of novocain with needling at the specific point in several groups of rabbits fed a high cholesterol diet for two or three weeks. Significant decreases in plasma and aortic cholesterol were obtained in the combined acupuncture group after one or two weeks of acupuncture. When the specific point was blocked by injection of a larger dose of novocain or the deep peroneal nerve was cut, then needling at the specific point lost its hypocholesterolemic effect. It is thus suspected that stimulation of sensory receptors of the deep peroneal nerve at the specific point may be the mechanism of the cholesterol-lowering action of acupuncture.

Acupuncture Therapy↗

Nuclear magnetic resonance studies of histone IV solution conformation.

The 220-MHz high-resolution proton magnetic resonance (PMR) spectrum of histone IV has been examined as a function of histone concentration, salt concentration, and pD. The hydrophobic C-terminal portion of the histone IV monomer appears to be largely PMR "invisible" indicating that this region of the polypeptide contains rigid secondary structure. Further loss of PMR resonance areas with increased histone IV concentration in neat D2O has been attributed to self-aggregation involving a monomer-dimer equilibrium. An equilibrium between the monomer and large aggregates, on the other hand, appears to dominate at NaCl concentrations above 0.01 M. pD studies reveal an abrupt increase in histone IV aggregation at pD smaller than 0.8 and precipitation of histone IV at pD values in the neighborhood of its isoelectric point, pD similar to 11.

Animals↗

Nuclear magnetic resonance studies on the solution conformation of histone IV fragments obtained by cyanogen bromide cleavage.

Two histone IV fragments obtained by cleavage at Met-84 by cyanogen bromide have been examined by proton magnetic resonance (PMR) spectroscopy as a function of temperature, peptide concentration, ionic strength, and pD. Sedimentation and gel electrophoresis studies on these peptides have also been carried out. The 220-MHz PMR spectrum of the N-peptide in both the high- and low-field regions was shown to be almost identical with that calculated for an extended coil N-peptide monomer. The calculated random coil and experimental C-peptide spectra, on the other hand, differ in many respects. Evidence was obtained for the presence of rigid secondary structure in the C-peptide. In addition, the Val, Leu, Ile CH3 resonance displays a prominent high-field satellite band which shifts downfield with increasing temperature. Sedimentation studies on the N-peptide reveal the formation of extremely large, remarkably homogeneous aggregates at ionic strengths larger than or equal to 0.01. The C-peptide, on the other hand, does not appear to form aggregates of sufficient size to be detectable in velocity sedimentation studies of a few hours duration. The relative area changes which have previously been noted in the PMR spectrum of histone IV with increasing ionic strength were also observed for the N-peptide but not the C-peptide. Interpretation of these relative area changes has been made in terms of the amino acid sequence of histone IV, and an effort was made to identify that segment of the polypeptide which undergoes secondary structural change with increasing ionic strength.

Amino Acids↗