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H Xu

Publications and source records attributed to H Xu.

At least 793 records · Page 44Linked to original sources
PubMed1988

A general method for the chromosomal amplification of genes in yeast.

The yeast retrotransposon Ty can be used to insert multiple copies of a gene at new sites in the genome. The gene of interest is inserted into a GALI-Ty fusion construct; the entire "amplification cassette" is then introduced into yeast on a high copy number plasmid vector. Transposition of the Ty element carrying the gene occurs at multiple sites in the genome. Two genes, a bacterial neomycin phosphotransferase gene and the yeast TRPl gene, were amplified in this way. Although the amplified genes were about 1 kilobase in length, they were amplified to about the same extent as a 40-base pair segment. The benefit of this "shotgun" approach is that amplification can be achieved in one set of manipulations.

DNA Transposable Elements↗
PubMed1988

The non-catecholamine-mediated electrophysiological effects of intravenous bethanidine sulfate on the immature mammalian heart.

In a preliminary study, we found that bethanidine sulfate had important electrophysiologic effects on the neonatal canine heart, specifically that bethanidine increased atrial effective and functional refractory periods. Other effects (increase in heart rate blood pressure and enhanced atrioventricular conduction) were thought to be due to a release of endogenous catecholamines. To investigate the non-catecholamine-mediated effects of bethanidine, we administered 10 mg/kg i.v. bethanidine to eight neonatal puppies ages 6-14 days pretreated with 0.6 mg/kg of propranolol and compared them with a control group of six neonates that received propranolol followed by a placebo. In the bethanidine group, the mean atrial effective and functional refractory periods increased significantly from 58 to 109 ms and from 108 to 185 ms, respectively (p less than 0.0001). Bethanidine also caused a decrease in resting heart rate (from 154 beats/min postpropranolol to 144 beats/min postbethanidine, p less than 0.002). These effects were not observed in the placebo group. Wenckebach periodicity during incremental atrial pacing did not change significantly. There was a modest increase in the ventricular refractory periods following bethanidine. Thus, the direct electrophysiologic effects of bethanidine in the neonate include a significant prolongation of atrial refractoriness and a decrease in sinus node automaticity. Ventricular refractory periods, while increased, did not show the dramatic prolongation exhibited by the atrium. The atrial specificity of bethanidine is unique and may prove useful in the treatment of supraventricular arrhythmias in the neonatal period.

Animals↗
PubMed1988

Isolation of Sperms from the Pollen Tubes of Flowering Plants during Fertilization.

Sperm cells have been isolated from pollen tubes growing in style segments of the dicotlyledon Rhododendron macgregoriae and the monocotyledon Gladiolus gandavensis by the in vivo/in vitro method at various stages of fertilization. Pollen tubes emerged from the cut end of the style into agar medium, and more than 95% contained sperm cells. Sperm cells were released from the pollen tubes by osmotic shock or by placing styles in wall-degrading enzymes: 0.5% macerozyme and 1% cellulase. The isolated sperms were ellipsoidal protoplasts of diameter about 2 x 3 micrometers in Gladiolus and about 3 x 4 micrometers in Rhododendron. After isolation, a proportion of the sperm cells occurred in pairs linked at one end by finger-like connections. The pairs of isolated sperms were dimorphic in terms of surface area and volume. By cutting the styles at various positions and times after pollination, the potential exists to detect changes in sperm gene expression associated with fertilization.

Journal Article↗
PubMed1988

Atrioventricular nodal function in the immature canine heart [corrected and republished article orginally printed in Pediatr Res 1987 Nov;22(5):616-20].

Previous studies have suggested that the atrioventricular nodal functional refractory period in the neonate is equal to or shorter than that of the ventricle, providing little or no protection to the ventricle against rapid atrial rates and allowing closely coupled atrial beats to fall within the ventricular vulnerable period. We evaluated atrioventricular node function in 21 mongrel neonatal puppies, 3-15 days old, and 15 adult dogs utilizing intracardiac His bundle recording and stimulation techniques. The mean atrioventricular nodal functional refractory period (173.1 +/- 20.0 ms) exceeded both the ventricular effective refractory period (139.5 +/- 14.3) and ventricular functional refractory period (163.3 +/- 14.5) in the neonates. Furthermore, the atrioventricular node was the site of limiting antegrade conduction in all neonates. No ventricular arrhythmias were induced by atrial extrastimulation in any of the neonates. The site of conduction delay during atrial extrastimulation was confined to the atrioventricular node in 15/16 neonates (94%) while 1/16 (6%) had combined nodal and infranodal delay. The neonates developed Wenckebach, at significantly faster heart rates than the adults, but both groups developed Wenckebach at approximately twice the resting heart rate. Retrograde conduction was a consistent finding in the neonates. However, antegrade Wenckebach occurred at a significantly faster heart rate than retrograde Wenckebach suggesting different functional properties. Our data suggest that in the neonatal canine, the atrioventricular node functional refractory period is longer than both the ventricular effective refractory period and ventricular functional refractory period. Furthermore, the degree of protection offered by the neonatal atrioventricular node to the ventricle appears to be comparable to that of the adult.

Animals↗
PubMed1988

In vivo demonstration of maturational changes of the chronotropic response to alpha-adrenergic stimulation.

In vitro studies suggest that neonates and adults may have different cardiac chronotropic responses to alpha-adrenergic stimulation. To investigate these differences in vivo, three groups of dogs were studied. Group I = 12 puppies, ages 3-7 days; group II = 12 puppies ages 8-15 days, and group III = seven adult dogs. Heart rate and blood pressure determinations were made in the control setting and then after combined beta-adrenergic and parasympathetic blockade (propranolol 0.6 mg/kg and bilateral vagotomies). Alpha-stimulation was then achieved with phenylephrine given in doses of 0.5, 1.0, and 10.0 micrograms/kg/min. A second, high dose of propranolol (1.0 mg/kg intravenously) was administered after the highest phenylephrine infusion dosage to assure complete beta-blockade. Finally, alpha-blockade was achieved with phentolamine (groups I and II: 0.5 mg intravenously; group III: 5.0 mg intravenously). An alpha-mediated positive chronotropic effect was observed in 42 and 100% of subjects in groups I and II, respectively, but never observed in the adults. Whereas alpha-blockade with phentolamine resulted in a large decrease in heart rate of all puppies (groups I and II), it had no effect on adults. Blood pressure responses were similar in all three groups. Thus, there are important maturational changes in the chronotropic response to alpha-adrenergic stimulation and blockade demonstrable in the intact neonatal canine.

Aging↗
PubMed1988

Multiple interactions of unsaturated fatty acids with opiate and ouabain binding sites and beta-adrenergic sensitive adenylate cyclase system.

The unsaturated fatty acids oleic, linoleic and arachidonic inhibited binding of ligands to the ouabain, opiate, and beta-adrenergic plasma membrane receptors. Low concentrations of fatty acids slightly increased the binding of ouabain to its binding sites. The effect of these fatty acids on beta-adrenergic sensitive adenylate cyclase was more complex. 0.2-0.3 mM fatty acids increased adenylate cyclase activity, while higher concentrations of arachidonic and linoleic acids, but not oleic acid, inhibited basal, beta-agonist- and NaF-stimulated activities in membranes of A431 and C6 cells. To evaluate which aspects of the unsaturated fatty acid molecules might be responsible for the observed effects, myristic acid, monoolein and taurodeoxycholic acid were studied. They also inhibited binding to the opiate receptor. Myristic acid, did not inhibit ouabain binding, binding to beta-receptor, nor adenylate cyclase activity. Monoolein, had no inhibitory effect on ouabain binding but behaved similar to oleic acid in the beta-receptor/adenylate cyclase system. Taurodeoxycholic acid inhibited binding to all three receptors as well as adenylate cyclase activity. We conclude that the effects of unsaturated fatty acids on ligand binding and adenylate cyclase activity are the result of their multiple interactions with various molecular processes rather than any unique property of long chain unsaturated fatty acids, per se.

Adenylyl Cyclases↗
PubMed1988

Regulation of biliary protein secretion in dogs.

The biliary secretion of protein in response to bile acids and other agents known to increase bile flow was examined in a chronic bile fistula dog model. Infusion of 25, 50, or 75 mumole/kg/hr sodium taurocholate after 3 hr of bile fistulization increased biliary protein output significantly by 52, 86, and 108% respectively compared to preinfusion values. A proportionate increase in biliary albumin output during taurocholate choleresis was demonstrated. Protein outputs during bile fistulization without taurocholate replacement were unchanged. The non-micelle-forming bile acid dehydrocholate markedly increased bile flow but did not change protein output. Similarly, the hormonal choleretics glucagon and secretin caused significant decreases in biliary protein concentration but no change in protein output. These data indicate a correlation between biliary protein secretion and bile acid-dependent bile flow. It is likely that regulation of certain proteins is dependent on the micelle-forming properties of bile acids.

Animals↗
PubMed1987

Binary mixtures of asymmetric phosphatidylcholines with one acyl chain twice as long as the other.

High-resolution differential scanning calorimetry and 31P NMR spectroscopy have been used to study aqueous phosphatidylcholine (PC) dispersions prepared from colyophilized mixtures of C(10):C(22)PC/C(22):C(12)PC of various molar ratios. These two lipid species are highly asymmetric but have a common structural feature; namely, one acyl chain in the fully extended conformation is about twice as long as the other. Our experimental results support two conclusions: (1) These two component lipids are miscible in all proportions in both gel and liquid-crystalline states. This type of system behaves as a nearly ideal mixture. Its calorimetric parameters are those expected on the basis of the mole fraction weighted average of the corresponding parameters for the pure components. (2) The component lipids appear to self-assemble, at T less than Tm, into a mixed interdigitated bilayer in excess water. In a mixed interdigitated bilayer, the short acyl chain of one asymmetric phosphatidylcholine on one side of the bilayer leaflet is apposed with the short acyl chain of another lipid molecule on the other side of the bilayer leaflet, while the longer acyl chain from each of the two leaflets crosses the entire hydrocarbon width of the bilayer. The fundamental packing unit, as well as the dynamic unit describing the axial rotator motion about the bilayer normal for this mixed interdigitated bilayer, is thus a dimer, whereas the packing unit assigned for the noninterdigitated bilayer such as C(16):C(16)PC lamellae is a monomer.

Calorimetry, Differential Scanning↗
PubMed1987

Scanning calorimetric study of fully hydrated asymmetric phosphatidylcholines with one acyl chain twice as long as the other.

The asymmetric C(18):C(10)PC molecules are known by X-ray diffraction to self-assemble, in excess water, into a lamellar structure known as the mixed interdigitated bilayer at T less than Tm. In this structure, the long C(18)-acyl chain is interdigitated fully across the entire hydrocarbon width of the bilayer, while the shorter C(10)-acyl chain, which is about half as long as the C(18)-acyl chain, packs end to end with a C(10)-acyl chain of another lipid molecule in the opposing bilayer leaflet. We have synthesized the following asymmetric phosphatidylcholines (PC's): C(16):C(9)PC, C(16):C(10)PC, C(18):C(10)PC, C(18):C(11)PC, C(20):C(11)PC, C(20):C(12)PC, C(22):C(12)PC, C(22):C(13)PC, C(8):C(18)PC, and C(10):C(22)PC. These 10 asymmetric phosphatidylcholines have a common characteristic; i.e., the length of the longer extended acyl chain is about twice as long as that of the shorter acyl chain. On the basis of the known lamellar structure of C(18):C(10)PC, we anticipate that these asymmetric phosphatidylcholines will also form mixed interdigitated bilayers. We have employed high-resolution differential scanning calorimetry (DSC) to investigate the thermotropic behavior of liposomes prepared from these asymmetric phosphatidylcholines. If our anticipation is correct, one would find that the thermodynamic data (Tm, delta H, or delta S) associated with the main thermal phase transitions of these asymmetric phosphatidylcholine dispersions will fit into a continuous curve as they are plotted as a function of the hydrocarbon width of the putative mixed interdigitated bilayer. Experimental data presented in this paper indeed bear this out. For comparison, a DSC study of multilamellar dispersions prepared from a series of saturated symmetric phosphatidylcholines has also been carried out.(ABSTRACT TRUNCATED AT 250 WORDS)

Calorimetry, Differential Scanning↗
PubMed1987

High-frequency deletion between homologous sequences during retrotransposition of Ty elements in Saccharomyces cerevisiae.

By following the fates of genetically marked Ty elements, we observed a very high frequency (80-90%) of deletion between directly repeated marker sequences during transposition. From blot hybridization analyses of Ty RNA and DNA species found in the Ty virus-like particles, we determined that the deletion events occurred during or immediately after reverse transcription of Ty RNA but before integration of Ty DNA. The results suggest that the Ty reverse-transcription machinery can recognize homologous sequences in the template. This capacity may be utilized in the replication and recombination processes of retrotransposons and retroviruses.

Chromosome Deletion↗
PubMed1987

The electrophysiologic effects of propafenone hydrochloride in the neonatal canine.

The electrophysiologic effects of the new antiarrhythmic agent, propafenone, were investigated in 10 mongrel canine neonates, ages 5 to 11 days. Utilizing standard His bundle recording and stimulation techniques, assessments of sinus and atrioventricular (AV) node function and atrial, AV nodal, and ventricular refractory periods were performed prior to (control) and after cumulative intravenous doses of 1, 2, and 4 mg/kg of propafenone. Propafenone depressed the spontaneous heart rate and prolonged the postatrial pacing recovery times. AV nodal function was depressed as manifested by Wenckebach periodicity occurring at slower pacing rates, increases in AV nodal conduction time, and increases in AV nodal refractoriness. Atrial and ventricular refractory periods were prolonged significantly in a dose-dependent fashion. Prolongation of the His-Purkinje conduction time occurred at the highest dose. Thus, propafenone exerts a generalized depressant effect on neonatal myocardial conduction and refractoriness which suggests that this agent may be useful in the therapy of atrial and ventricular dysrhythmias in the immature heart.

Animals↗
PubMed1987

Demonstration and characterization of two classes of cardiac glycoside binding sites to rat heart membrane preparations using quantitative computer modeling.

Cardiac glycoside binding to rat heart membrane preparations was measured by rapid filtration technique. The binding data were analyzed using quantitative computer analysis. The experimental results using [3H]-ouabain as the labeled ligand were consistent with a model in which cardiac glycoside specific binding occurs at two independent classes of sites. The high affinity sites were characterized by a dissociation constants of 40 nM, 50 nM, and 61 nM for ouabain, digoxin and digitoxin, respectively, with a binding capacity of 1.3 pmoles/mg protein. The lower affinity sites for ouabain were characterized by dissociation constants of 2.3 microM, 67 nM and 71 nM for ouabain, digoxin and digitoxin, respectively, with a binding capacity of 3 pmoles/mg protein. Potassium ions inhibit [3H]-ouabain binding in a dose dependent manner with an IC50 of 500 microM. Quantitative computer modelling indicated that potassium inhibits ouabain binding at both binding sites.

Animals↗