Biomedical subjects

L Huang

Publications and source records attributed to L Huang.

At least 667 records · Page 37Linked to original sources

PubMed1990

[Sedative-hypnotic effect of caulis Polygoni multiflori decoction].

The sedative-hypnotic effect of Caulis Polygoni multiflori decoction (CPMD) was studied by the rolling cylinder method in mice and by the somnopolygraphic recording method in rats. The result from the rolling method in mice showed that CPMD (9g/kg) did have synergic action with the subthreshold hypnotic dose of sodium pentobarbital (20mg/kg). The result from the somnopolygraphic recording experiment showed that the immediate hypnotic effect of an effective dose of CPMD (20g/kg) was basically similar to that of diazepam (5mg/kg) used under the same experimental condition, chiefly an increase in the duration of slow wave sleep phase (SWS) and a decrease in the duration of paradoxical sleep phase (PS). By the end of a three-day session in dose of twice a day the hypnotic effect appeared more prominent and the reduction of SWS latency became significant.

Animals↗

PubMed1989

Highly efficient immunoliposomes prepared with a method which is compatible with various lipid compositions.

Monoclonal antibody was conjugated to N-glutaryl-phosphatidylethanolamine in the presence of octylglucoside by using N-hydroxysulfosuccinimide as a carboxyl-activation reagent. The conjugated antibody was then incorporated into liposomes by a simple dialysis method. The method is mild and is compatible with various lipid compositions of the liposomes. We have prepared immunoliposomes containing a lung endothelium-specific monoclonal antibody and showed excellent target binding (approximately 75% injected dose) of the immunoliposomes in mouse. Immunoliposomes can be prepared to contain other acidic lipids such as phosphatidylserine and various amounts of cholesterol. The presence of 20% or more cholesterol in liposomes resulted in high level of target binding. We have used in these experiments a new radioactive lipid-phase marker, 111In-DTPA-SA, which was very stable in vivo. The halflife of clearance in mouse exceeded 3 weeks.

Animals↗

PubMed1989

Immunoliposomes with different acid sensitivities as probes for the cellular endocytic pathway.

By combining dioleoylphosphatidylethanolamine (DOPE) with oleic acid (OA), palmitoylhomocysteine (PHC) or dipalmitoylsuccinylglycerol (DPSG) we have prepared pH-sensitive liposomes with different acid sensitivities. DOPE/OA liposomes are the most acid sensitive, while DOPE/DPSG liposomes are the least acid sensitive. Incubation of DOPE/OA liposomes with mouse L929 cells reduces the pH-sensitivity of these liposomes by altering the lipid composition. Using diphtheria toxin fragment A as a marker for cytoplasmic delivery, we find that the delivery kinetics of pH-sensitive immunoliposomes closely correlates with the modified acid sensitivities of the liposomes. Immunoliposomes encounter pH 6-6.2 with a t1/2 of 5-15 min after internalization. By contrast, acidification of the endosomes to pH 5.0 takes longer (t1/2 approximately 25 min). We also used a whole cell null point technique (Yamishiro and Maxfield (1987) J. Cell Biol. 105, 2713-2721) to directly determine the average pH encountered by the endocytosed immunoliposomes. We find that acidification determined by the null point method proceeds less rapidly than that estimated from DTA delivery data. This is likely due to the fact that the measured DTA delivery is done by those liposomes which first arrive at the endosomes with sufficient acidity. Our data suggests that DOPE/PHC immunoliposomes deliver at the early endosome while DOPE/DPSG immunoliposomes deliver at the late endosomes. The DOPE/OA immunoliposomes, with the altered composition and acid sensitivity, deliver with a kinetics intermediate between the other two immunoliposomes. Thus, pH-sensitive liposomes represent useful probes for studying the kinetics of endosome acidification.

Animals↗

PubMed1989

Highly efficient DNA delivery mediated by pH-sensitive immunoliposomes.

We have previously shown that pH-sensitive immunoliposomes can mediate a target-specific delivery of plasmid DNA to tumor cells grown in a mouse model [Wang, C.-Y., & Huang, L. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7851-7855]. The efficiency of delivery in terms of the target cell transformation frequency has now been characterized for both short- and long-term gene expression in a tissue culture system. Herpes simplex virus thymidine kinase (TK) gene was used as a reporter gene. It was placed under the control of the promoter for the rat phosphoenolpyruvate carboxykinase gene, which contains a cAMP regulatory element. Therefore, the expression of the exogenous gene in the target cell, mouse Ltk- cells, can be regulated by cAMP drugs. The plasmid DNA was encapsulated in liposomes using a detergent dialysis method. The efficiency of gene delivery was optimized with respect to the time course and dose of liposome-associated DNA. The existence of antibody of the liposomes was essential for the maximal level of DNA delivery. Delivery was also dependent on the lipid composition of the liposome. The pH-sensitive lipid composition gave 8-fold higher efficiency than the corresponding pH-insensitive composition. The transformation efficiency of the target cell also depended on the regulation of gene expression; cells incubated with dibutyryl-cAMP and theophylline showed a much higher level of transformation frequency than cells incubated without the drugs. When all liposome and incubation parameters are optimized, the Ltk- cells showed a 47% efficiency for the short-term transformation, and 2% for the long-term transformation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

The role of protein-linked oligosaccharide in the bilayer stabilization activity of glycophorin A for dioleoylphosphatidylethanolamine liposomes.

The importance of the protein-linked carbohydrates for the stabilization of dioleoylphosphatidylethanolamine (DOPE) bilayers has been investigated using glycophorin A, the major sialoglycoprotein of the human erythrocyte membrane, as a stabilizer. Two major types of glycophorin, differing in the sialic acid content, were used in the study. Type MM contains 19.2 +/- 2.5 sialic residues per molecule of glycophorin, and type NN contains 10.8 +/- 1.2. Type MM could stabilize DOPE bilayers at 0.5 mol%, whereas type NN was unable to do so even at 1 mol%. The importance of the sialic acid content to the stabilization activity of glycophorin was further confirmed by the observation that the neuraminidase-treated type MM showed a lower stabilization activity than the untreated type. Since type NN had no stabilizing activity, we attempted to couple a trisaccharide, NeuNAc----Gal----Glc, to type NN by reductive amination. 2.5 +/- 0.8 saccharide chains were added per molecule of type NN. The trisaccharide-attached type NN showed a greater stabilization activity than the parent type NN molecule, indicating again that the sialic acid content of the stabilizer molecule determines the stabilization activity. Addition of wheat-germ agglutinin (WGA), which binds to the sialic acid residues of a glycoprotein, to type MM-stabilized liposomes caused rapid aggregation and destabilization of liposomes, resulting in leakage of an entrapped marker, calcein. The aggregation increased with increasing amount of the lectin; however, the leakage rate was maximum at an optimum concentration of WGA. These results are discussed in terms of the role of sialic acid in the interfacial hydration and charge repulsion which determines the DOPE bilayer stability.

Carbohydrate Sequence↗

PubMed1989

Monoclonal antibody targeting of liposomes to mouse lung in vivo.

A monoclonal antibody (MoAb), 273-34A, specifically binds to an epitope expressed almost exclusively on capillary endothelial cells of the lung. Within 15 min after i.v. injection, approximately 80 to 85% of the injected radioiodinated MoAb 273-34A accumulates in the lung. Approximately 90 to 95% of the recovered dose is found in the lung for up to 1 week postinjection. Ratios of MoAb 273-34A to a nonspecific, irrelevant MoAb 135-14 are 250 to 285 times higher in the lung than in the serum. When 273-34A was coupled with palmitic acid and incorporated into liposomes, the amount of 125I-labeled liposomes recovered per g of tissue was 12 times higher in the lung than in the liver at 15 min postinjection, and 22 times higher at 5 h postinjection. At 24 h postinjection the amount of liposomes per gram of lung tissue was still 6.0 times the amount per gram of liver tissue. Liposomes conjugated to MoAb 273-34A locate in the lung 20 and 15 times better than do liposomes conjugated to the nonspecific MoAb 135-14 at 15 min and 24 h postinjection, respectively. The results indicate that this immunoliposome system could be used as a model for enhanced drug delivery to the lung. The potential use for delivering anticancer drugs for therapy of lung tumors is discussed.

Animals↗

PubMed1989

Use of a quaternary ammonium detergent in liposome mediated DNA transfection of mouse L-cells.

Sonicated liposomes composed of dioleoylphosphatidylethanolamine (DOPE) and a quaternary ammonium detergent (dodecyl-, tetradecyl-, or cetyl-trimethylammonium bromide) mediates functional transfer of pSV2 CAT plasmid DNA to mouse L929 fibroblasts. Successful transfection was determined by assaying for chloramphenicol acetyltransferase activity in cell lysates collected 40 h after exposure to the lipid-DNA complexes. Liposomes prepared with the quaternary ammonium detergents were less toxic than the free detergents at the same concentrations and were more efficient in their delivery of the plasmid DNA to the cells. Analysis of the three detergents in combination with the lipid showed that cetyltrimethylammonium bromide was least toxic to the cells. This detergent, at a minimal concentration of 20 mol% in DOPE, allowed for stable liposome preparations and efficient transfection. Optimal efficiency of transfection occurred with 30 micrograms of DNA. Further increases in the DNA concentration caused a decrease in the transfection efficiency, perhaps due to charge repulsions between the liposomes now saturated with negatively charged DNA and the negatively charged cell surface. The transfection activity of the liposome was limited by its cytotoxicity at high liposome concentrations. These results are compared with that of the Lipofectin, another positively charged liposome preparation which is commercially available. Although the overall transfection activity of the liposome containing the quaternary ammonium detergent is somewhat lower than that of the Lipofectin, it may serve as an inexpensive and convenient alternative.

Animals↗

PubMed1989

Small, but not large, unilamellar liposomes composed of dioleoylphosphatidylethanolamine and oleic acid can be stabilized by human plasma.

Small unilamellar liposomes, composed of dioleoylphosphatidylethanolamine (DOPE) and oleic acid (OA), prepared by sonication, were incubated in the presence of human plasma at 37 degrees C. The release of entrapped calcein after 8-h incubation was about 15% in plasma, compared with about 70% in phosphate-buffered saline under the same conditions. In contrast, dioleoylphosphatidylcholine (DOPC)/OA liposomes under the same conditions release about 70% in plasma and only 10% in PBS. Total release of calcein from the DOPE/OA liposomes was observed in a PBS solution containing bovine serum albumin, and the release was completely blocked by preincubation of the liposomes with plasma. These results indicate that the unstable DOPE/OA liposomes are stabilized by incubation with plasma. The stabilization process was very fast, being completed within 1 min. Only relatively small liposomes (d less than or equal to 200 nm) were completely stabilized by plasma; larger liposomes were progressively less stabilizable. SDS-polyacrylamide gel electrophoresis of liposomes which had been incubated with plasma and then washed indicated that several proteins were tightly associated with liposomes. Using liposomes containing [14C]OA, it was found that about 70% of the original OA was extracted after 1-h incubation with human plasma at 37 degrees C. Thin-layer chromatographic analysis of the plasma-treated liposomes showed the presence of the plasma lipids in the liposomes. These results suggest that liposomes composed of DOPE/OA are stabilized by protein and/or lipid components from human plasma and that the composition of the liposomes is altered. The mechanism of stabilization is discussed in terms of the surface pressure of small vesicles with a high degree of curvature.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗

PubMed1989

Structure and function relationship of phosphatidylglycerol in the stabilization of the phosphatidylethanolamine bilayer.

Differential scanning calorimetry was used to examine the structure-function relationship of the phospholipids on the L alpha-phase stabilization of phosphatidylethanolamine (PE). Phosphatidylglycerol (PG) was chosen as a model stabilizer. Dielaidoylphosphatidylethanolamine (DEPE) was mixed with various PGs to study the effects of (i) chain length, (ii) chain unsaturation, and (iii) chain number of the stabilizer on the L alpha-phase stabilization. At low concentrations of stabilizer, both bilayer stabilization and destabilization were observed. Phase separations also were seen, as revealed by split peaks of the L beta----L alpha transition; these were particularly prone to occur in the destabilization cases. When saturated PGs were compared, shorter chains (C12:0 and C14:0) promoted bilayer stabilization whereas longer chains (C16:0 and C18:0) promoted bilayer destabilization. Unsaturated PG with larger hydrophobic volumes (C18:2) favored bilayer destabilization, relative to unsaturated PG with smaller hydrophobic volumes (C18:1). Lyso-PG (C14:0) showed higher bilayer stabilization activity than their double-chain counterparts. Thus, at low concentrations of stabilizer, the acyl chain composition plays a vital role in bilayer-phase stabilization. However, at higher concentrations (greater than or equal to 8 mol %), all PGs become active bilayer stabilizers. This is probably because the increased head-group hydration becomes the dominant factor in the stabilization. The effect of acyl chain composition of the stabilizer was also studied by using small unilamellar vesicles composed of dioleoylphosphatidylethanolamine (DOPE). Fluorescence quenching of calcein entrapped in liposomes was used to monitor the stability of the liposomes. Similar acyl chain effects on liposomal stabilization were obtained.(ABSTRACT TRUNCATED AT 250 WORDS)

Calorimetry, Differential Scanning↗

PubMed1989

Characterization of plasma-stabilized liposomes composed of dioleoylphosphatidylethanolamine and oleic acid [published errtum appears in Biochem Biophys Res Commun 1989 Sep 29;163(3):1539].

We have previously reported that small unilamellar liposomes (d less than or equal to 200 nm) composed of dioleoylphosphatidylethanolamine and oleic acid can be stabilized by incubating with normal human plasma (Liu and Huang, Biochemistry 1989, in press). The stabilized liposomes were very stable even under relatively harsh conditions such as extreme pH, high salt and trypsin treatment. Fluorescence depolarization of diphenylhexatriene showed that the stabilized liposome had a high microviscosity in the lipid core, which did not decrease even after the majority of proteins were removed by trypsin. These data suggest that plasma proteins inserted into the lipid bilayer are probably responsible for the stabilization activity. After i.v. injection into mouse, stabilized liposomes showed a relatively low affinity to liver and spleen as compared to a conventional liposome composition.

Animals↗

PubMed1989

Role of cholesterol in the stability of pH-sensitive, large unilamellar liposomes prepared by the detergent-dialysis method.

Large unilamellar liposomes prepared by an octyl glucoside-dialysis method were examined for stability at 37 degrees C in the presence or absence of human plasma, using the release of the entrapped calcein as a fluorescence marker. The liposomes were acid-sensitive as they were composed of dioleoylphosphatidylethanolamine, oleic acid and cholesterol. The stability of the liposomes in the absence of plasma was significantly enhanced with increasing cholesterol content. However, the maximal calcein release at pH 5 decreased linearly with increasing cholesterol content of the liposome, indicating that cholesterol had reduced the acid-sensitivity of the liposomes. In the presence of human plasma, calcein release exhibited a biphasic behavior with a fast (plasma-sensitive) and a slow (plasma-resistant) component. Inclusion of cholesterol in the liposomes resulted in an increased proportion of the plasma-resistant component. Liposomes pretreated with human plasma, after removal of excess plasma and the released calcein by gel-filtration, showed a remarkable stability both in the presence and absence of human plasma. The acid-sensitivity of the plasma-treated liposomes with 40% cholesterol was the same as that of the untreated liposomes. These results are discussed in terms of the mechanism by which these liposomes deliver their contents to the cytoplasm of the cells via the endocytic pathway, a known biological activity of the type of liposome described here.

Blood↗

PubMed1989

Regulation of 5-Aminolevulinic Acid (ALA) Synthesis in Developing Chloroplasts : II. Regulation of ALA-Synthesizing Capacity by Phytochrome.

When dark-grown cucumber (Cucumis sativus L.) seedlings previously exposed to white light for 20 hours were returned to darkness, the ability of isolated chloroplasts to synthesize 5-aminolevulinic acid dropped by approximately 70% within 1 hour. The seedlings were then exposed to light, and the synthetic ability of the isolated chloroplasts was determined. Restoration of the synthetic capacity was promoted by continuous white or red light of moderate intensity. Intermittent red light was also effective. Blue and far-red light did not restore the synthetic capability. Blue light given after a red pulse did not enhance the effect of the red light. Far-red light given immediately after each red pulse prevented the stimulation due to intermittent red light. Restoration of the biosynthetic activity by in vivo light treatments was inhibited by cycloheximide indicating the requirement for translation on 80 S ribosomes for the in vivo light response. These findings suggest that the majority of the plastidic 5-aminolevulinic acid synthesis is under phytochrome regulation.

Journal Article↗

PubMed1989

Regulation of 5-aminolevulinic Acid synthesis in developing chloroplasts : I. Effect of light/dark treatments in vivo and in organello.

Intact chloroplasts isolated from greening cucumber (Cucumis sativus L. var Beit Alpha) cotyledons regenerated protochlorophyllide (Pchlide) in the dark with added cofactors from either exogenous glutamate or endogenous substrates. No other intermediates of the chlorophyll biosynthetic pathway accumulated. When inhibitors of 5-aminolevulinic acid (ALA) dehydratase were added, the Pchlide that failed to form was replaced by an excessive amount of ALA. When greening seedlings were returned to the dark, ALA-synthesizing activity in the isolated chloroplasts decreased dramatically and recovered if the dark-treated seedlings were again exposed to continuous white light prior to chloroplast isolation. Both the decline and the recovery of ALA-synthesizing activity were complete in approximately 50 minutes. Changes in chloroplast structure during in vivo light to dark and dark to light transitions (as evidenced by electron microscopy) were much slower. Exposing isolated chloroplasts from dark-treated seedlings to short white flashes before incubation transformed nearly all the endogenous Pchlide, but hardly stimulated ALA synthesis, suggesting that Pchlide does not act as a feed-back inhibitor on ALA synthesis. Chloroplasts isolated from dark-treated tissue did not form Pchlide from glutamate when incubated in the dark with added cofactors; moreover, the endogenous Pchlide did not turn over in organello. However, these chloroplasts did synthesize Pchlide from added ALA at the normal rate and synthesized ALA from glutamate at a reduced, but still significant, rate. Mg chelation was not affected by in vivo dark treatment.

Journal Article↗

PubMed1989

[Distributions and gene frequencies of PGM1 subtype, EsD, GLO1, AK, ADAand 6-PGD in 20 races of China].

The distributions and gene frequencies of PGM1, subtype EsD, GLO1, AK, ADA and 6-PGD of 4171 healthy donors in 20 races of China were detected using gel electrophoresis. Some isoenzymes such as PGM1, PGM1 subtype, EsD and GLO1 have better distribution and higher discriminating power than the others. Sixty-eight samples with PGM1(6) gene were observed in 12 races. The frequency in zhuang population is the highest among 20 races. The phenotype of PGM1 6-1 is up to 4.15%. The band of PGM1(6) of all 68 samples is in the same position in the gel. In general speaking, the gene frequencies of EsD in most races of southern part of China are higher than those of northern part. Phenotype EsD 2 in Hani, Lisu, Dai, Naxi, She, Zhuang, Dong, Miao are over 15%, Hani is up to 32.4%. The gene frequency of GLO1 in Tajik and Uygur are 0.297 and 0.2112 respectively. The others range from 0.0714 to 0.1527. In Qiang it is 0.0583. The gene frequencies of AK1. ADA1 and 6-PGDA are very high in 20 races, and with very low discriminating power.

Adenosine Deaminase↗

PubMed1989

[Clinical characteristics of silent myocardial ischemia and the effect of nifedipine treatment].

The characteristics of ischemic episodes in exercise test and daily activities were observed in a silent myocardial ischemia (SMI) group and an anginal group (23 patients each). 15 patients in the SMI group were treated with nifedipine. In exercise test, the time of onset of ischemia was earlier and the ischemic threshold was lower in SMI group. During daily activities, the frequency of SMI was high. The heart rate just before onset of SMI was lower than the mean heart rate in 24-hour Holter monitoring. The highest frequency of SMI was found between 5 AM and 12 noon. Postinfarction patients had a higher frequency and a longer duration of SMI than noninfarction patients. The frequency and duration of SMI decreased in the 15 patients treated with nifedipine in SMI group. It is concluded that silent ischemic episodes were frequent and occurred easily. They might be associated with poor prognosis in CAD patients. Nifedipine was effective in reducing the frequency and duration of SMI in our patients.

Aged↗

PubMed1989

Anterior ischemic optic neuropathy--recurrent attacks in one eye in a bilateral case.

Subsequent involvement of the fellow eye is common in anterior ischemic optic neuropathy, but recurrent episodes in one eye are extremely rare. Our hypertensive patient had one eye attacked nine years before and two episodes in the second eye within ten days of each other. Visual-field examination revealed progressing field loss, and fluorescein angiography demonstrated an enlarged disc hypofluorescence area.

Fluorescein Angiography↗

PubMed1989

[Studies of the total synthesis of (-)-ranunculin].

The synthesis of (-)-ranunculin from mannitol is described. The oxidation of 1,2,5,6-diacetone d-mannitol with NaIO4 or lead tetraacetate gave 2,3-O-isopropylidene-D-glyceraldehyde with the desired alpha-carbon-(R)-configuration. The Wittig reaction of isopropylidene-D-glyceraldehyde with (C6H5)3P = CHCOOCH3 gave a mixture of Z and E methyl-3-(2,2-dimethyl-1,3-dioxo-lan-4-yl)-2-propenoate with a Z:E ratio of 85:15. The dependence of Z:E ratio upon the solvent present was studied. On careful treatment of the cis isomer with 0.1 mol/L HCl, hydrolysis of the ketal took place along with cyclization to give the optically active aglycone with the expected stereochemistry, which was followed by Koenigs-Knorr reaction to give (-)-ranunculin tetraacetate. Since the glycoside is sensitive to acid and base, hydrolysis was carried out with strongly acidic cation exchange resin which yielded (-)-ranunuculin smoothly. The overall yield from mannitol with all the intermediates purified amounted to 15%.

Furans↗

PubMed1988

beta-Galactosidase-induced destabilization of liposome composed of phosphatidylethanolamine and ganglioside GM1.

A novel type of liposome bilayer destabilization catalyzed by the enzyme, beta-galactosidase, is described. Unsaturated phosphatidylethanolamine (PE), an HII-phase-forming lipid, does not form stable liposomes at physiological temperature and pH. However, stable unilamellar liposomes can be prepared by mixing PE with a minimum of 5 mol% ganglioside GM1, a micellar-phase-forming lipid. Treatment of these GM1/PE liposomes with beta-galactosidase induces a rapid leakage (3-6 min) of the entrapped fluorescent dye, calcein. The studies indicate that liposome destabilization is the result of catalytic degradation of GM1, rather than a stoichiometric binding of GM1 by beta-galactosidase. Kinetic data indicate that the destabilization takes place via liposome collision. This simple, rapid method of liposome destabilization by beta-galactosidase will be useful in designing a liposome-based signal amplification mechanism for assays involving enzymes.

Drug Stability↗