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J S Liang

Publications and source records attributed to J S Liang.

At least 19 recordsLinked to original sources
PubMed1998

The acute phase response in apolipoprotein A-1 knockout mice: apolipoprotein serum amyloid A and lipid distribution in plasma high density lipoproteins.

In plasma, the bulk of apoSAA, a positive acute phase reactant protein, is transported in high density lipoproteins (HDL), especially HDLH (apoA1-rich HDL). In this study we tested whether apoA1 deficiency would adversely affect apoSAA concentration and lipid distribution in mouse plasma lipoproteins. Acute phase response (APR) was induced in C57BL/6J (apoA1+/+) and apoA1-knockout mice (apoA1-/-) by a subcutaneous injection of silver nitrate. The APR increased cholesterol concentrations in LDL of apoA1-/- mice and apoA1+/+ mice in a like manner. In contrast to apoA1+/+ mice, concentrations of cholesterol, phospholipids and proteins in both HDLL (1.063<d<1.103 g/ml) and HDLH (1.103<d<1.21 g/ml) were significantly increased by the APR in apoA1-/- mice. Total concentration of plasma apoSAA and its distribution in lipoprotein fractions was similar in both APR groups. The bulk of plasma apoSAA was contained in HDL and not in VLDL or LDL even when the HDL concentration was low. In apoA1-/- mice, HDLL and HDLH contained more apoSAA than in apoA1+/+ mice. These results indicate that apoA1-/- mice are not deterred from mounting an apoSAA response similar to apoA1+/+ mice and that apoA1-rich HDL particles are not necessary for apoSAA transport in the plasma.

Acute-Phase Reaction↗
PubMed1998

Catabolism of lipid-free recombinant apolipoprotein serum amyloid A by mouse macrophages in vitro results in removal of the amyloid fibril-forming amino terminus.

Serum amyloid A fibrils are formed when the normally rapid catabolism of the acute-phase reactant apolipoprotein serum amyloid A (apoSAA) is incomplete; thus amyloidosis may be viewed as a condition of dysregulated proteolysis. There is evidence that apoSAA is dissociated from plasma high-density lipoprotein (HDL) prior to fibril formation. The objective of this study was to investigate degradation of lipid-free apoSAA by tissue macrophages derived from amyloid-susceptible CBA/J mice in vitro. Peritoneal macrophages derived from untreated (normal) mice converted apoSAA (12 kDa) to a single 4 kDa C-terminal peptide while splenic macrophages converted apoSAA to 10, 7 and 4 kDa C-terminal peptides and a 4 kDa peptide that lacked the C- and N-terminal regions. Similar patterns of proteolysis occurred when peritoneal and splenic macrophages from amyloidotic CBA/J mice were used. Conditioned medium prepared from peritoneal, but not splenic macrophages, degraded apoSAA. Specific sites of cleavage indicated activity of cathepsin G- and elastase-like neutral proteases. The data indicate that lipid-free apoSAA can be degraded by secreted or cell-associated neutral proteases that are generated by macrophages to yield peptides that lack fibrillogenic potential.

Amino Acid Sequence↗
PubMed1997

Evidence for local production of acute phase response apolipoprotein serum amyloid A in Alzheimer's disease brain.

Acute phase serum amyloid A (A-apoSAA), but not constitutive apoSAA (C-apoSAA), was identified by Western blotting experiments in brain protein extracts from eight of nine patients with Alzheimer's disease (AD), one with a brain tumor and one with multiple sclerosis. A-apoSAA was not detected in six subjects with Pick's or Lewy Body disease or three other non-AD brain specimens. Apolipoprotein A-I and albumin were not found in any of the brain protein extracts. A-apoSAA mRNA was detected in AD brain by reverse transcription-polymerase chain reaction (RT-PCR). These data suggest that apoSAA is locally produced in AD brain and that investigation of the neuroinflammatory effects of this injury specific apolipoprotein is warranted.

Aged↗
PubMed1996

The fibril forming region of the beta-amyloid precursor differs from that of the amyloid A precursor in its interaction with lipids.

Since the amyloid A (AA) precursoir, serum amyloid A (apoSAA), has been shown to bind cholesterol (C) in the AA fibril forming region, we investigated the interaction of the beta-amyloid precursor protein (AbetaPP) and beta-amyloid (Abeta) peptide with C and phosphatidyl choline (PC) by measuring changes in binding to microtiter wells at physiological pH and ionic strength. While either C or PC inhibited AbetaPP binding to the same extent that C inhibited apoSAA binding, neither C nor PC had any effect on binding of the Abeta peptide, although antibodies to Abeta1-40 did block binding. The binding of (125)I-Abeta1-40 and (125)I-AbetaPP was inhibited by apoE3 and apoE4, but not by either apoSAA or bovine serum albumin. Bound (125)I-AbetaPP was partially released into medium containing C, PC, apoE3, apoE4, or antibodies to AbetaPP. Our results indicate that AbetaPP but not Abeta peptide can be retained in solution in the presence of C and PC and suggest that this failure to interact with lipids may account for the greater insolubility of Abeta fibrils than AA fibrils.

Amyloid beta-Protein Precursor↗
PubMed1996

Polymorphism of acute-phase serum amyloid A isoforms and amyloid resistance in wild-type Mus musculus czech.

Until CE/J mice and their offspring were characterized as amyloid-resistant, all mice were thought to be amyloid-susceptible to multiple injections of azocasein or a single injection of silver nitrate following administration of amyloid enhancing factor. We now report, for the first time, that wild-type Mus musculus czech and F1 hybrids bred by crossing M. musculus czech with amyloid-susceptible CBA/J mice are also amyloid resistant. Based on the derived amino acid sequences of two serum amyloid A (SAA) cDNA clones, we describe two unusual SAA gene isoforms in M. musculus czech, one of which differs from four previously characterized acute-phase apoSAA isoforms at several amino acid residues. Our findings support the hypothesis that protection against amyloid fibril formation in wild-type M. musculus czech mice and their offspring is linked to apoSAA gene mutations (molecular motif).

Amino Acid Sequence↗
PubMed1996

Degradation of serum amyloid A in amyloid-susceptible and amyloid-resistant mouse strains.

Degradation of serum amyloid A (apoSAA) by resident peritoneal cells (RPCS) and conditioned medium (CDM), prepared with RPCS, from amyloid-susceptible CBA/J mice, amyloid-resistant CE/J mice and their amyloid-resistant CBA/J x CE/J F1 progeny was investigated in vitro. Serum amyloid A was derived from murine acute phase (AP) plasma and associated with high density lipoprotein (HDL). Degradation of apoSAA by RPCS and CDM from CBA/J mice was complete while degradation by RPCS and CDM from CE/J mice did not occur. Degradation of apoSAA by RPCS and CDM from CBA/J x CE/J F1 hybrid mice was indistinguishable from that by RPCS and CDM from the CBA/J parent. Intermediate fragments were not detected with either RPCS or CDM from CBA/J mice or CBA/J x CE/J F1 hybrid mice. Degradation of apoSAA was inhibited by phenylmethanylsulfonyl fluoride (PMSF) indicating that the enzyme, secreted into the fluid phase, was a serine esterase. Unlike apoSAA, HDL-associated apoA-1 remained intact. It was thus concluded that while selective degradation of HDL-associated apoSAA (apoSAA-HDL) by RPCS from the CBA/J and CE/J mice was significantly different, the genetic study did not support the hypothesis that there was direct linkage between impaired degradation of apoSAA-HDL in the CE/J mouse strain and protection against amyloid fibril formation. As amyloid resistance in CBA/J x CE/J F1 hybrid mice is not attributable to failure to express the amyloidogenic isoform apoSAA2, the study supports the original hypothesis that amyloid resistance may be linked to expression of apoSAAcej.

Amyloid↗
PubMed1996

Amino terminal region of acute phase, but not constitutive, serum amyloid A (apoSAA) specifically binds and transports cholesterol into aortic smooth muscle and HepG2 cells.

The human apoSAA proteins comprise both acute phase (apoSAA1, apoSAA2) and constitutive (apoSAA4) isoforms; all are expressed in human atherosclerotic lesions as well as in liver. Recombinant acute phase apoSAA binds cholesterol with an affinity of approximately 170 nM and enhances cholesterol uptake by HepG2 cells (J. Lipid Res. 1995. 36:37-46). In the present study, we sought to define the region of acute phase apoSAA involved in cholesterol binding and to investigate the ability of constitutive apoSAA4 to bind cholesterol. Binding of [3H]cholesterol to apoSAAp was inhibited by unlabeled cholesterol (1-100 nM), but not significantly by vitamin D and estradiol. Direct binding of acute phase, but not constitutive, apoSAA to the surfaces of polystyrene microtiter wells was strongly diminished in the presence of cholesterol. The ability of apoSAAp to bind cholesterol was inhibited by antibodies to human apoSAA1 and to peptide 1-18 of apoSAA1. There was only slight inhibition of cholesterol binding by antibodies to peptide 40-63, and no inhibition by antibodies to peptides spanning regions containing amino acid residues 14-44 and 59-104. [3H]cholesterol uptake by neonatal rabbit aortic smooth muscle and HepG2 cells was enhanced by a synthetic peptide corresponding to amino acids 1-18 of hSAA1, but not by peptides corresponding to amino acids 1-18 of hSAA4. [3H]cholesterol uptake by HepG2 cells was slightly increased by a peptide corresponding to amino acids 40-63 of hSAA1. These findings suggest that apoSAA modulates the local flux of cholesterol between cells and lipoproteins during inflammation and atherosclerosis.

Amino Acid Sequence↗
PubMed1995

Recombinant human serum amyloid A (apoSAAp) binds cholesterol and modulates cholesterol flux.

During acute inflammation, the serum amyloid A (apoSAA) proteins apoSAA1 and apoSAA2 are transiently associated with high density lipoproteins (HDL) in concentrations of as much as 1000-fold more than their concentrations during homeostasis; however, their effect on HDL function is unclear. Recombinant apoSAAp, a hybrid of the closely related human apoSAA1 and apoSAA2 isoforms, was found to exhibit a high affinity for cholesterol. The adsorption of apoSAAp to polystyrene microtiter wells at physiological pH, temperature, and salt concentration was inhibited and reversed by cholesterol. ApoSAAp, to a greater extent than apoA-I, albumin, or fetal bovine serum, enhanced diffusion of cholesterol from HDL across a membrane that retained molecules > 3.5 kDa. Cholesterol from 25 nM to 125 microM inhibited binding of [3H]cholesterol to 167 nM apoSAAp. A cholesterol binding assay was developed to determine the dissociation constant for binding of [3H]cholesterol to apoSAAp; Kd = 1.7 +/- 0.3 x 10(-7) M and the maximum binding capacity (Bmax) is 1.1 +/- 0.1 mol/mol. After binding cholesterol, the apparent size of apoSAAp as determined by gel filtration on Sephacryl S-100 was increased from 12 to 23 kDa. ApoSAAp enhanced free [14C]cholesterol uptake from tissue culture medium by HepG2 cells, an effect that was dose dependent and blocked by polyclonal antibodies to human apoSAA1 and apoSAA2. ApoSAAp, unlike apoA-I, was taken up from serum-free medium by HepG2 cells and appeared to be degraded by cell-associated enzymes. Unlike peritoneal exudate cells, human HepG2 hepatoma cells do not secrete an enzyme that degrades apoSAAp. These results suggest that apoSAA can potentially serve as a transient cholesterol-binding protein.

Adsorption↗
PubMed1993

m1-toxin.

The venom of the Eastern green mamba from Africa, Dendroaspis angusticeps, contains a number of toxins which block the binding of 3H-antagonists to genetically-defined m1 and m4 muscarinic acetylcholine receptors. Most of the anti-muscarinic activity of the venom is due to the presence of a newly-isolated toxin, "m1-toxin", which has 64 amino acids and a molecular mass of 7361 Daltons. At present m1-toxin is the only ligand which is known to be capable of fully blocking m1 receptors without affecting m2-m5 receptors. It binds very rapidly, specifically and pseudoirreversibly to the extracellular face of m1 receptors on cells, in membranes or in solution, whether or not the primary receptor site is occupied by an antagonist. Bound toxin can either prevent the binding and action of agonists or antagonists, or prevent the dissociation of antagonists. The toxin is useful for identifying m1 receptors during anatomical and functional studies, for recognizing and stabilizing receptor complexes, and for occluding m1 receptors so that other receptors are more readily studied.

Acetylcholine↗
PubMed1993

Differences in the plasma transport and tissue concentrations of tocopherols and tocotrienols: observations in humans and hamsters.

Certain aspects of tocopherol and tocotrienol absorption, plasma transport, and tissue distribution were examined in humans and hamsters. Plasma transport differed in that tocopherols were found primarily in low density lipoprotein and high density lipoprotein in association with plasma surface components, whereas tocotrienols disappeared from plasma with chylomicron clearance. In keeping with transport by triglyceride-rich lipoproteins, tocotrienols were deposited in conjunction with triglycerides in the adipose tissue of hamsters. In hamsters, tocopherols were the only tocol readily detected in all tissues, except adipose during tocotrienol supplementation. In fasting humans, the plasma tocotrienol concentration was not significantly increased after tocotrienol supplementation, whereas the platelet concentration of delta-tocotrienol doubled. Furthermore, tocotrienol intake did not appear to modulate the plasma cholesterol concentration in normolipemic hamsters. Thus, the transport, tissue concentration, and relative biologic function of tocopherol and tocotrienol appear somewhat disparate and possibly unrelated.

Animals↗
PubMed1993

Use of m1-toxin as a selective antagonist of m1 muscarinic receptors.

m1-Toxin is the only ligand which is known to bind specifically to the extracellular face of genetically defined m1 muscarinic receptors; it binds pseudoirreversibly. A variety of studies were performed to evaluate the usefulness of m1-toxin as a selective antagonist of m1 receptors. Exposure of slices of the rat cerebral cortex to m1-toxin in physiological buffer blocked the subsequent binding of 1.0 nM [3H]pirenzepine to m1 receptors in the slices. The toxin also blocked 70% of carbachol-stimulated turnover of radiolabeled inositol phosphates in hippocampal slices. Autoradiographs showed that m1-toxin bound to sections of once-frozen tissue and blocked the binding of [3H]quinuclidinyl benzilate to regions of the rat brain rich in m1 receptors. The toxin blocked the binding of [3H]antagonists to pure m1 receptors on the surface of living Chinese hamster ovary cells, but did not block intracellular receptors. In membrane preparations from the rat cortex and hippocampus the toxin blocked the binding of [3H] antagonists to m1 receptors quantitatively and selectively, but had no effect on binding sites for [3H]nicotine. Subsaturating amounts of the toxin bound to m1 receptors in membranes at 4 degrees C in less than 30 sec. Low concentrations of m1-toxin blocked m1 receptors in solution in digitonin but had no effect on separate preparations of pure m2, m3, m4 or m5 receptors. Thus m1-toxin appears to be a very useful antagonist for m1 receptors in intact tissue, on isolated cells, in membranes and in solution, in a variety of media.

Animals↗
PubMed1993

Stable allosteric binding of m1-toxin to m1 muscarinic receptors.

m1-Toxin was found to slow the dissociation of [3H]N-methyl-scopolamine (NMS) and [3H]pirenzepine from m1 muscarinic receptors expressed in the membranes of Chinese hamster ovary cells. When toxin-NMS-receptor complexes were formed in membranes and then dissolved in digitonin, or when these complexes were formed in solution, the toxin completely stopped the dissociation of [3H]NMS for 6 hr at 25 degrees C. Toxin-receptor complexes formed in membranes or in solution were also highly stable in solution at 25 degrees, as shown by the ability of the toxin to prevent the binding of [3H]quinuclidinyl benzilate (QNB). [3H] QNB-receptor complexes were equally stable, whereas unliganded soluble receptors lost most of their ability to bind QNB within an hour. Toxin-receptor complexes could be partially dissociated by incubation at 37 degrees in the presence of digitonin and [3H]QNB, and the freed receptors were then labeled. These results demonstrate that m1-toxin binds allosterically and pseudoirreversibly to m1 receptors, and that the toxin can stabilize the outward-facing pocket of m1 receptors which contains and binds competitive antagonists. The allosteric nature of the binding of m1-toxin should prove to be useful for such unusual purposes as stabilizing the binding of readily reversible and/or nonselective ligands specifically to m1 receptors, for purifying labeled or unlabeled receptors by affinity techniques which recognize the toxin, for recognizing receptors with genetically or biochemically altered primary binding sites, and for stabilization of the native conformation of m1 receptors for structural studies.

Allosteric Site↗
PubMed1993

Purification and properties of m1-toxin, a specific antagonist of m1 muscarinic receptors.

The venom of the Eastern green mamba from Africa, Dendroaspis angusticeps, was found to block the binding of 3H-quinuclidinyl benzilate to pure m1 and m4 muscarinic ACh receptors expressed in Chinese hamster ovary cells. The principal toxin in the venom with anti-m1 muscarinic activity was purified by gel filtration and reversed-phase HPLC. This toxin has 64 amino acids, a molecular mass of 7361 Da, and an isoelectric point of 7.04. Its cysteine residues are homologous with those in curare-mimetic alpha-neurotoxins, and with those in fasciculin, which inhibits AChE. At low concentrations the toxin blocked m1 receptors fully and pseudoirreversibly while having no antagonist activity on m2-m5 receptors; the toxin is therefore named "m1-toxin." At higher concentrations m1-toxin interacted reversibly with m4 receptors, and half of the toxin dissociated in 20 min at 25 degrees C. The affinity of m1-toxin is therefore much higher for m1 than for m4 receptors. By comparison with m1-toxin, pirenzepine has sixfold higher affinity for m1 than for m4 receptors. Autoradiographs of muscarinic receptors in the rat brain demonstrated that m1-toxin blocked the binding of 2 nM 3H-pirenzepine only in regions known to bind m1-specific antibodies. Thus, m1-toxin is a much more selective ligand than pirenzepine for functional and binding studies of m1 muscarinic receptors.

Amino Acid Sequence↗
PubMed1992

Immunoglobulin A against viral capsid antigen of Epstein-Barr virus and indirect mirror examination of the nasopharynx in the detection of asymptomatic nasopharyngeal carcinoma.

To evaluate the efficacy of population screening for early stage nasopharyngeal carcinoma (NPC) in southern China, the authors recruited 42,048 and 10,402 apparently healthy subjects residing in a high incidence and a low incidence area, respectively; all subjects were between the ages of 30 and 59 years. The subjects' serum specimens were tested for immunoglobulin (Ig) A antibody against viral capsid antigen (IgA/VCA) of Epstein-Barr virus (EBV). Of the subjects from the high incidence area, 2823 were found to be seropositive. In follow-up, they had yearly examinations of the nasopharynx by indirect mirror with or without biopsy; 41 were found to have histologically confirmed asymptomatic NPC during the first 2 years of follow-up. The tumors in most of these cases were localized and were at earlier stages than tumors of symptomatic cases of NPC seen in the same region before the screening. The yearly indirect mirror examination of the nasopharynx seems to have effectively identified most of the tumors at the stage of asymptomatic disease. The risk of harboring NPC was found to be different among the different sex and age subgroups of seropositive individuals. By limiting such screening to those who are at exceedingly high risk, the cost of the screening can be kept within the spending of the public health authority, and the effectiveness of the screening also is improved.

Adult↗
PubMed1979

Isolation and partial synthesis of a new metabolite of medroxyrogesterone acetate.

3 alpha-Hydroxy-17-acetoxy-6 alpha-methyl-5 beta-pregnan-20-one (IIIa) has been isolated from urine of patients receiving medroxyprogesterone acetate (MPA). It was characterized by partial synthesis from MPA by catalytic reduction with palladium-charcoal to 17-acetoxy-6 alpha-methyl-5 beta-pregnan-3,20-dione (IV) and reduction of the latter with sodium borohydride. The isolation of 6 beta, 17,21-trihydroxy-6 alpha-methyl-pregn-4-ene-3,20-dione (IIc) is reported for the first time. The 17- and 21-monoacetates of this compound have been isolated and characterized earlier by other investigators. 7 alpha-3H-Medroxyprogesterone acetate was administered to 4 subjects by intravenous and intramuscular injections and by mouth. The ring A saturated metabolite IIIa was excreted in 0.1% to 4.0% of the administered dose; the highest excretion was after the intravenous dose and lowest after oral ingestion. 6 beta, 17,21-Trihydroxy-6 alpha-methylpregn-4-ene-3,20-dione (IIc) and its 17- and 21-monoacetates were excreted in about 5% of the doses in all subjects. No increase in 6 beta-hydroxylation was observed in the patient treated with o,p'-DDD,2,2-bis(2-chlorophenyl, 4'-chlorophenyl)-l,1-dichloroethane.

Adult↗