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Biomedical subjects

S Y Liu

Publications and source records attributed to S Y Liu.

At least 55 records · Page 3Linked to original sources
PubMed1997

Modification of heart sarcolemmal phosphoinositide pathway by lysophosphatidylcholine.

Although lysophosphatidylcholine (lyso-PtdCho) accumulates in the sarcolemmal (SL) membrane and alters its function during myocardial ischemia and diabetic cardiomyopathy, the effects of lyso-PtdCho on SL signalling processes have not yet been investigated. The present study was carried out to examine the actions of lyso-PtdCho on the rat heart SL membrane enzymes involved in the phosphoinositide pathway. Different lyso-PtdCho species (10 to 200 microM) inhibited the activities of both phosphatidylinositol kinase and phosphatidylinositol-4-phosphate kinase in the SL membrane in a concentration-dependent manner. The inhibitory potency of lyso-PtdCho compounds for phosphatidylinositol kinase was lyso-PtdCho plasmalogen > 1-oleoyl-lyso-PtdCho > 1-stearoyl-lyso-PtdCho > 1-palmitoyl-lyso-PtdCho, and that for phosphatidylinositol-4-phosphate kinase was lyso-PtdCho plasmalogen > 1-oleoyl-lyso-PtdCho > 1-palmitoyl-lyso-PtdCho > 1-stearoyl-lyso-PtdCho. The inhibitory effect of lyso-PtdCho on phosphatidylinositol-4-phosphate kinase was greater than that on phosphatidylinositol kinase. Lyso-PtdCho structural analogues, such as phosphatidylcholine, lysophosphatidic acid, lysophosphatidylethanolamine, L-alpha-glycerophosphate, oleate and phosphorylcholine, did not affect the phosphoinositide kinases, suggesting that the intact structure of lyso-PtdCho was required for the inhibition of the kinases. The detrimental action of lyso-PtdCho on PtdIns kinase was potentiated by acidosis. Unlike Ca2+, ATP (0.1 and 4 mM) increased lyso-PtdCho-induced deactivation of the kinases. Both enzyme activities were found to be depressed in the ischemic-reperfused or diabetic hearts. None of the tested lyso-PtdCho species altered phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) hydrolysis by SL phospholipase C. These results indicate that accumulation of lyso-PtdCho in the SL membrane under pathological conditions may diminish the availability of the PtdIns(4,5)P2 substrate for the production of second messengers by receptor-linked phospholipase C.

1-Phosphatidylinositol 4-Kinase↗
PubMed1997

Heparin-binding properties of selenium-containing thioredoxin reductase from HeLa cells and human lung adenocarcinoma cells.

Mammalian selenocysteine-containing thioredoxin reductase (TR) isolated from HeLa cells and from human lung adenocarcinoma cells was separated into two major enzyme species by heparin-agarose affinity chromatography. The low-affinity enzyme forms that were not retained on heparin agarose showed strong crossreactivity in immunoblot assays with anti-rat liver TR polyclonal antibodies, whereas the high-affinity enzyme forms that were retained by the heparin column were not detected. Both low and high heparin-affinity enzyme forms contained FAD, were indistinguishable on SDS/PAGE analysis, and exhibited similar catalytic activities in the NADPH-dependent DTNB [5,5'-dithiobis(2-nitrobenzoate)] assay. The C-terminal amino acid sequences of 75Se-labeled tryptic peptides from lung adenocarcinoma low- and high heparin-affinity enzyme forms were identical to the predicted C-terminal sequence of human placental TR. These two determined peptide sequences were -Ser-Gly-Ala-Ser-Ile-Leu-Gln-Ala-Gly-Cys-Secys-(Gly). Occurrence of the Se-carboxymethyl derivative of radioactive selenocysteine in the position corresponding to TGA in the gene confirmed that UGA is translated as selenocysteine. The presence of cysteine followed by a reactive selenocysteine residue in this C-terminal region of the protein may explain some of the unusual properties of the mammalian TRs.

Adenocarcinoma↗
PubMed1997

Selenophosphate synthetase: enzyme labeling studies with [gamma-32P]ATP, [beta-32P]ATP, [8-14C]ATP, and [75Se]selenide.

Selenophosphate synthetase catalyzes a reaction in which ATP and selenide are converted to H3SeP03, H3P04, and AMP in a 1:1:1 ratio. Selenophosphate is derived from the gamma phosphoryl group and orthophosphate from the beta phosphoryl group of ATP. In the absence of selenide, a slow reaction in which ATP is converted quantitatively to 2 H3P04 and AMP occurs. Labeling experiments carried out to detect a putative enzyme-bound pyrophosphate intermediate in the overall reaction showed that up to 0.6 equivalent of the 32P label from [gamma-32P]ATP was bound to protein under enzyme turnover conditions, but only a negligible amount of 32P from [beta-32P]ATP was present. Thus, no Enz-PP intermediate was present in a detectable amount under the experimental conditions used. Isolated enzyme samples contained 75Se from 75Se-labeled selenide and [14C]AMP from [8-14C]ATP in amounts similar to the bound 32P from [gamma-32P]ATP, suggesting that two of the final products, selenophosphate and AMP, were the radioactive compounds detected in these experiments.

Adenosine Triphosphate↗
PubMed1997

A non-radioactive and two radioactive assays for selenophosphate synthetase activity.

Selenophosphate synthetase catalyzes the formation of monoselenophosphate (SePO3(3-)) from ATP and selenide (reaction 1). [formula: see text] In one assay frequently used, [8-14C]AMP formation from [8-14C]ATP is estimated after separation of the nucleotides by thinlayer chromatography. An alternative non-radioactive assay in which the AMP product is estimated using AMP deaminase is described. The highly oxygen-labile selenophosphate product can be estimated in an assay employing [gamma-32P]ATP. The 32P-labeled selenophosphate is converted to [32P]orthophosphate by treatment with iodine and estimated after removal of residual [32P]ATP on charcoal.

Adenosine Monophosphate↗
PubMed1997

Preventive effects of traditional Chinese (Kampo) medicines on experimental osteoporosis induced by ovariectomy in rats.

Preventive effects by traditional Chinese (Kampo) medicines, Unkei-to, Hachimi-jio-gan, and Juzen-taiho-to, on the progress of bone loss induced by ovariectomy in rats were investigated for a period of 49 days. The bone mineral density (BMD) of tibia in ovariectomized (OVX) rats decreased by 20% from those in sham-operated (Sham) rats, with the decrease completely inhibited by the administration of any one of these Kampo medicines or 17beta-estradiol. From scanning electron microscopic (SEM) analyses, the surface of a trabecular bone of tibia in OVX rats had a porous or erosive appearance, whereas that of the same bone in Sham rats was composed of fine particles. The administration of three Kampo medicines and 17beta-estradiol to OVX rats preserved the fine particle surface of the trabecular bone. These results strongly suggest that any of these three gynecological Kampo medicines is as effective as 17beta-estradiol in preventing the development of bone loss induced by ovariectomy in rats.

Absorptiometry, Photon↗
PubMed1997

The potential for immunoconjugates in lymphoma therapy.

Contemporary combination chemotherapy offers curative treatment for 30% to 50% of patients with advanced-stage, aggressive non-Hodgkin's lymphomas (NHL). However, conventional therapies cure few patients with indolent lymphomas or relapsed lymphomas of any histology. Myeloablative chemoradiotherapy with bone marrow or stem cell transplantation can provide pro-longed disease-free survival for a minority (20% to 50%) of patients with relapsed NHL, but new treatment approaches are clearly needed. In recent years, several groups of investigators have provided preliminary evidence suggesting that monoclonal antibodies (mAbs), in unmodified form or conjugated to toxins, drugs, or radioisotopes, may offer another effective therapeutic modality for patients with relapsed lymphomas. This article reviews current immunotherapy of NHL using antibody immunoconjugates.

Animals↗
PubMed1997

Anthraquinones in Rheum palmatum and Rumex dentatus (Polygonaceae), and phorbol esters in Jatropha curcas (Euphorbiaceae) with molluscicidal activity against the schistosome vector snails Oncomelania, Biomphalaria and Bulinus.

Hot water extracts of Rheum plamatum and Rheum dentatus (from China) showed molluscicidal activity against the snails Oncomelania hupensis, Biomphalaria glabrata and Bulinus globosus, which are vectors of Schistosoma japonicum, S. mansoni and S. haematobium, respectively. Activity was correlated with antraquinones which were identified by HPLC: rhein and chrysophanol-anthron were most active (> 50% dead snails after 2 days in a 0.03% solution). Molluscicidal activity was intermediate with Rheum-emodin and physcion and was not detectable with cinnamic acid or Aloe-emodin. The snail O. hupensis tended to be more sensitive for several compounds than B. glabrata. Extracts of Jatropha curcas seeds (from Mali) showed molluscicidal activity against both B. glabrata and O. hupensis, the latter being the more sensitive snail. The activity was associated with phorbol esters extracted from Jatropha oil. Of the pure phorbol esters tested, 4 beta-phorbol-13-decanoate killed both snail species at a concentration of 0.001% (10 p.p.m). As Jatropha is locally grown in Mali for other purposes, it might potentially be exploited for schistosomiasis control.

Animals↗
PubMed1997

[Synthesis and biological activity of substance P analogs].

Substance P (SP) and its three analogs were synthesized through solid-phase procedure. The fully-protected peptide-resin was cleaved with anhydrous HF.--SH of Cys in [Cys5,9] SP(4-11) was protected with Acm, then deprotected through Iodine oxidation method, the disulfide bridge cyclization was formed subsequently. The crude product was purified on C18 RP-HPLC. The results of guinea-pig isolated ileum test (GPI) showed that the potency of the analogs was as follows: [AcGln5, Pro9] SP(5-11) > SP > SP(4-11) > [Cys5,9] SP(4-11). They induced GPI to contract very fast and showed the characteristics of tachykinin. On the writing test in mice, the results showed: [Cys5,9] SP(4-11) > SP > SP(4-11) > [AcGln5, Pro9] SP(5-11). The biological tests showed that the SP analogs have some extent of selectivity.

Animals↗
PubMed1996

Evaluation of a monoclonal antibody-based sandwich-ELISA for detection of circulating Schistosoma japonicum antigen (Sj31/32) in an endemic area of China.

A sandwich-ELISA specific for 31/32 kDa schistosome proteins was used to trace infections in a Chinese population living in a low endemicity area for schistosomiasis japonica. The study population included 304 persons for whom records were available from 1992 onwards. In 1994, 2 individuals were positive in the miracidial hatching assay performed with 30 g of stools. The ELISA gave positive results for 8 individuals, including these 2 with active infections. Of the remaining 6 ELISA-positive, 5 had excreted eggs and received praziquantel in 1993 and may have harboured residual worms in 1994. It is concluded that this ELISA is able, in a low prevalence situation, to identify the few infected individuals.

Adolescent↗
PubMed1996

Effects of a rhubarb (Rhei rhizoma) solution and its fractions on the formation of calcium phosphate precipitates.

Inhibitory effects of a rhubarb (Rhei rhizoma) solution and its fractions on the formation of calcium phosphate precipitates were studied. The rhubarb solution inhibited both the amorphous calcium phosphate (ACP) formation and the rate of hydroxyapatite (HAP) transformation, and extended the induction time. When the solution was fractionated using membrane filters, a filtrate with the molecular weight between 3 and 10 kDa (with 2/3 recovery of polyphenols) was found to be responsible for both the ACP formation and the extension of the induction time. Another filtrate with the molecular weight below 3 kDa (with 1/3 recovery of polyphenols) may be responsible for the inhibition of both the ACP formation and the rate of HAP transformation, and the extension of the induction time. When the extract of rhubarb was fractionated using a Sephadex LH-20 column chromatography, fraction IV greatly inhibited the formation of calcium phosphate precipitates, while fractions I, II and III slightly inhibited that reaction. Our finding suggests that fraction IV may contain useful substance(s) for the prevention of oral calcium phosphate precipitation (calculus formation). However, strong calcium chelating properties would limit the concentration that could be safely employed.

Analysis of Variance↗
PubMed1996

Purification and cloning of a thermostable xylose (glucose) isomerase with an acidic pH optimum from Thermoanaerobacterium strain JW/SL-YS 489.

An unusual xylose isomerase produced by Thermoanaerobacterium strain JW/SL-YS 489 was purified 28-fold to gel electrophoretic homogeneity, and the biochemical properties were determined. Its pH optimum distinguishes this enzyme from all other previously described xylose isomerases. The purified enzyme had maximal activity at pH 6.4 (60 degrees C) or pH 6.8 (80 degrees C) in a 30-min assay, an isoelectric point at 4.7, and an estimated native molecular mass of 200 kDa, with four identical subunits of 50 kDa. Like other xylose isomerases, this enzyme required Mn2+, Co2+, or Mg2+ for thermal stability (stable for 1 h at 82 degrees C in the absence of substrate) and isomerase activity, and it preferred xylose as a substrate. The gene encoding the xylose isomerase was cloned and expressed in Escherichia coli, and the complete nucleotide sequence was determined. Analysis of the sequence revealed an open reading frame of 1,317 bp that encoded a protein of 439 amino acid residues with a calculated molecular mass of 50 kDa. The biochemical properties of the cloned enzyme were the same as those of the native enzyme. Comparison of the deduced amino acid sequence with sequences of other xylose isomerases in the database showed that the enzyme had 98% homology with a xylose isomerase from a closely related bacterium, Thermoanaerobacterium saccharolyticum B6A-RI. In fact, only seven amino acid differences were detected between the two sequences, and the biochemical properties of the two enzymes, except for the pH optimum, are quite similar. Both enzymes had a temperature optimum at 80 degrees C, very similar isoelectric points (pH 4.7 for strain JW/SL-YS 489 and pH 4.8 for T. saccharolyticum B6A-RI), and slightly different thermostabilities (stable for 1 h at 80 and 85 degrees C, respectively). The obvious difference was the pH optimum (6.4 to 6.8 and 7.0 to 7.5, respectively). The fact that the pH optimum of the enzyme from strain JW/SL-YS 489 was the property that differed significantly from the T. saccharolyticum B6A-RI xylose isomerase suggested that one or more of the observed amino acid changes was responsible for this observed difference.

Aldose-Ketose Isomerases↗
PubMed1996

Cloning, sequencing, and expression of the gene encoding a large S-layer-associated endoxylanase from Thermoanaerobacterium sp. strain JW/SL-YS 485 in Escherichia coli.

The gene (xynA) encoding a surface-exposed, S-layer-associated endoxylanase from Thermoanaerobacterium sp. strain JW/SL-YS 485 was cloned and expressed in Escherichia coli. A 3.8-kb fragment was amplified from chromosomal DNA by using primers directed against conserved sequences of endoxylanases isolated from other thermophilic bacteria. This PCR product was used as a probe in Southern hybridizations to identify a 4.6-kb EcoRI fragment containing the complete xynA gene. This fragment was cloned into E. coli, and recombinant clones expressed significant levels of xylanase activity. The purified recombinant protein had an estimated molecular mass (150 kDa), temperature maximum (80 degrees C), pH optimum (pH 6.3), and isoelectric point (pH 4.5) that were similar to those of the endoxylanase isolated from strain JW/SL-YS 485. The entire insert was sequenced and analysis revealed a 4,044-bp open reading frame encoding a protein containing 1,348 amino acid residues (estimated molecular mass of 148 kDa).xynA was preceded by a putative promoter at -35 (TTAAT) and -10 (TATATT) and a potential ribosome binding site (AGGGAG) and was expressed constitutively in E. coli. The deduced amino acid sequence showed 30 to 96% similarity to sequences of family F beta-glycanases. A putative 32-amino-acid signal peptide was identified, and the C-terminal end of the protein contained three repeating sequences 59, 64, and 57 amino acids) that showed 46 to 68% similarity to repeating sequences at the N-terminal end of S-layer and S-layer-associated proteins from other gram-positive bacteria. These repeats could permit an interaction of the enzyme with the S-layer and tether it to the cell surface.

Amino Acid Sequence↗
PubMed1996

Inhibitory effects of 1-hydroxyethylidene-1,1-bisphosphonate and Chinese traditional (kampo) medicines on calcification of the heart and tongue in DBA/2NCrj mice.

The effects of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) and two Chinese traditional (Kampo) medicines, Dai-saiko-to and Saiko-ka-ryukotsu-borei-to on spontaneous soft tissue (heart and tongue) mineralization in DBA/2NCrj mice were studied. These agents were given orally for 4 weeks to DBA/2NCrj mice. After 2 weeks of administration in the heart, 0.006 and 0.03% (w/v) HEBP decreased calcium content by 90 and 30%, respectively, while 0.27 and 2.7 mg/ml Dai-saiko-to reduced calcium content by 30 and 45%, respectively. Saiko-ka-ryukotsu-borei-to (0.27 mg/ml) reduced both calcium and phosphorus content by 50 and 35%, respectively. However, their inhibitory effects on the heart were not observed after 4 weeks of administration. The compounds delayed the onset of increases of bulk calcium and phosphorus content. In the tongue, at 4 weeks, 0.006 and 0.03% (w/v) HEBP reduced calcium content by 30 and 45%, respectively, while two Kampo medicines (at both concentrations used) significantly reduced the content of calcium (by 27-79%) and phosphorus (by 24-32%). These results strongly suggest that two Kampo medicines as well as HEBP may be useful in preventing and curing soft tissue calcification.

Animals↗
PubMed1996

Alpha 1-adrenergic receptors in the mammalian cochlea.

The presence of alpha 1-adrenergic receptors in the mammalian cochlea has previously been suggested by physiological experiments using antagonists specific to the receptor. However, the characteristics of adrenergic receptors in the cochlea have not been described. By employing [3H]-prazosin, high affinity, specific binding sites with characteristics of alpha 1-adrenergic receptors have now been identified and characterized in the chinchilla cochlea. Analysis of the specific [3H]-prazosin binding indicates that prazosin binds to a single class of high-affinity sites with a dissociation constant, kd, of 2.9 x 10(-9) M and a maximum number of binding sites, Bmax, of 30 fmol/mg dry tissue. Furthermore, the binding characteristics suggest that these receptors may be related to the microvasculature of the cochlea. These results provide a rational basis for the observed actions of alpha-adrenergic drugs on the auditory system.

Adrenergic alpha-Antagonists↗