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Biomedical subjects

C S Liu

Publications and source records attributed to C S Liu.

At least 109 records · Page 6Linked to original sources

Influence of Mg2+ and pH on n.m.r. spectra and radioligand binding of inositol 1,4,5-trisphosphate.

We and others have shown that the binding of Ins(1,4,5)P3 to its receptor is pH-sensitive and can be inhibited by Mg2+. In the present study we have used 1H- and 31P-n.m.r. spectroscopy to study whether these effects results from increased ionization of Ins(1,4,5)P3 and a direct interaction with Mg2+ respectively. Under near-physiological conditions of ionic strength (100 mM-KCl), three ionizable groups were observed. The pH titration curve of the 1-phosphate was monophasic, with a pKa of 6.3. The titration curves of the 4- and 5-phosphates were biphasic, suggesting that these groups interact; the pKa values for the 4-phosphate determined by 31P-n.m.r. were 5.7 and 7.8, and for the 5-phosphate they were 5.3 and 7.9. 1H- and 31P-n.m.r. measurements suggest that Mg2+ binds weakly to Ins(1,4,5)P3 at physiological pH. Mg2+ non-competitively inhibited binding of Ins(1,4,5)P3 to its receptor in rat cerebellum and bovine adrenal cortex. Inhibition curves for rat cerebellum at pH 7.1 and 8.5, and also for bovine adrenal cortex at pH 8.5, appeared to be monophasic, with IC50 values (concn. of displacer giving 50% inhibition of specific binding) of 214 microM, 572 microM and 9.1 mM respectively. Scatchard analysis revealed that Mg2+ inhibited binding of Ins(1,4,5)P3 to bovine adrenal cortex at pH 8.5 in a non-competitive manner. Our results suggest that the previously reported pH-sensitivity of the binding of Ins(1,4,5)P3 may be caused by ionization of the phosphate groups in positions 4 and 5, and that the ability of Mg2+ to inhibit the binding of Ins(1,4,5)P3 is not mediated by direct chelation but through a site located on, or close to, the Ins(1,4,5)P3 receptor. Inhibition by Mg2+ is pH-sensitive and can vary at least 10-fold between tissues, suggesting possible receptor heterogeneity. Mg2+ may exert an important regulatory control on the release of Ca2+ by Ins(1,4,5)P3.

Adrenal Cortex↗

Purification and characterization of an antiplatelet peptide, arietin, from Bitis arietans venom.

By means of Fractogel TSK-50, CM-Sephadex C-50 column chromatography, gel filtrations on Sephadex G-75 and Sephacryl S-200 columns and reverse-phase HPLC, an antiplatelet peptide, arietin, was purified from venom of Bitis arietans. Arietin was shown to be an Arg-Gly-Asp-containing peptide with a NH2-terminus, Ser-Pro-Pro-Val-Cys-Gly-Asn-Lys- (Mr 8500). Arietin dose-dependently inhibited aggregation of human platelet suspension stimulated by ADP, thrombin, collagen and U46619 with IC50, 1.3-2.7.10(-7) M, while it had no effect on the initial shape changes and only slightly affected ATP release of platelets caused by thrombin and collagen. Arietin also blocked platelet aggregation in platelet-rich plasma and whole blood, and inhibited thrombin-induced clot retraction of platelet-rich plasma. Furthermore, arietin (6.5.10(-8) M) completely blocked the fibrinogen-induced aggregation of elastase-treated platelets, indicating that arietin interferes with the fibrinogen binding to fibrinogen receptors on platelet membranes. In conclusion, arietin, an Arg-Gly-Asp-containing peptide, inhibits platelet aggregation probably through the blockade of fibrinogen binding to the activated platelets.

Adenosine Diphosphate↗

The amino acid sequence and properties of an edema-inducing Lys-49 phospholipase A2 homolog from the venom of Trimeresurus mucrosquamatus.

Three phospholipase A2 enzymes or homologs were purified from the venom of Trimeresurus mucrosquamatus (Taiwan habu). The most abundant one was found to be a phospholipase homolog without enzyme activity, and its complete amino acid sequence was determined using oligopeptide fragments derived from digestion by endopeptidases Glu-C, Asp-N, Lys-C and alpha-chymotrypsin, and by means of gas-phase sequencing. The sequence revealed that the protein belonged to the Lys-49 family of snake venom phospholipase A2. This protein's function was characterized as edema-inducing. The Lys-49 protein has the potential to bind membrane phospholipid and Ca2+ (Kd = 1.6 x 10(-4) M) as shown by ultraviolet difference spectra; however, the catalytic site appeared to be inactive and the edematous response was independent of the protein's hydrolytic activity. Mast cells and platelets were shown to be subject to activation by the Lys-49 protein.

Amino Acid Sequence↗

Systemic amyloidosis presenting as splenic tumor.

We report a case of amyloidosis manifesting as a splenic tumor and pleural effusion. Only one case of a splenic amyloid tumor has been reported in the literature, but its radiological findings were not described. Therefore, we present the computed tomographic (CT), sonographic, and angiographic features of splenic amyloid tumor.

Amyloidosis↗

A randomised double blind trial comparing the treatment of episcleritis with topical 2-(2-Hydroxy-4-methylphenyl) Aminothiazole Hydrochloride 0.1% (CBS 113A) and placebo.

A randomised double blind trial of 2-(2-Hydroxy-4-methylphenyl) Aminothiazole Hydrochloride 0.1% (CBS 113A) versus placebo was carried out in 43 eyes with episcleritis. Our results show that CBS 113A is effective in reducing the signs of conjunctival and episcleral inflammation in mild episcleritis within the first week of administration of the drug (Day 3: Total Score p = 0.0013, Conjunctival Injection p = 0.017, Episcleral Injection p = 0.0018 and Day 7: Total Score p = 0.01, Conjunctival Injection p = 0.014, Episcleral Injection p = 0.027). CBS 113A was not effective against severe episcleritis. No significant side effects were found apart from a stinging sensation. There was no effect on intraocular pressure. The potential use of this new drug is discussed.

Adult↗

[The chromatographic-mass spectrometric analysis and detection of anabolic steroids in human urines and metabolic study].

Nineteen anabolic steroids were separately administered to healthy men. Anabolic steroid metabolites were isolated and extracted from the collected urine samples following an integrated procedure. The main metabolic pathways of these drugs were made clear after the investigation by GC-MS. Based on the obtained chromatographic-mass spectrometric data of anabolic steroids, a method for large scale and routine analysis of anabolic steroids was set up.

Adult↗

[Investigation on the metabolism of dehydrochloromethyltestosterone in man].

A GC-MS method for the investigation on the metabolism of dehydrochloromethyltestosterone (DHCMT) in man is described. Steroids in human urine were adsorbed on a macroporous XAD-2 resin, eluted with methanol, hydrolyzed with glucuronidase, extracted and concentrated for derivatization and then analyzed with GC-MS. DHCMT and its 7 metabolites were detected in urine samples 8 to 30 h after oral administration. Their structures and the variation of their concentration in urine were determined and the possible metabolic pathways of DHCMT were proposed on the basis of GC-MS feature of its metabolites. The specific metabolites and characteristic ions for screening DHCMT positive urine were chosen. Furthermore, the influence of the sample pretreatment methods on GC-MS results has been studied.

Gas Chromatography-Mass Spectrometry↗

[GC-MS analysis of norethandrolone and its metabolites in man].

Steroids in human urine were adsorbed on a macroporous XAD-2 resin, eluted with methanol, hydrolyzed with glucuronidase, extracted and concentrated for TMS derivatization and then analyzed with GC-MS. Norethandrolone (N) and its 9 metabolites were detected in urine samples 9-35 h after oral administration. Met-2 and Met-7 could be detected even in 84 h urine sample. The structures of 7 metabolites were elucidated and the variations of their concentration in urine were determined. The specific metabolites and characteristic ions for screening N positive urine were chosen. This method is sensitive enough; the detection limit of N was 10 ng per ml urine, i.e. 10 ppb.

Gas Chromatography-Mass Spectrometry↗

Triflavin, an antiplatelet Arg-Gly-Asp-containing peptide, is a specific antagonist of platelet membrane glycoprotein IIb-IIIa complex.

Triflavin, an antiplatelet peptide containing Arg-Gly-Asp, purified from Trimeresurus flavoviridis venom, inhibits aggregation of human platelets stimulated by a variety of agonists. It blocks aggregation through interference with fibrinogen binding to its specific receptor on the platelet surface membrane in a competitive manner, but it has no apparent effect on intracellular events, such as thromboxane B2 formation, phosphoinositides breakdown and intracellular Ca2+ mobilization of thrombin-activated platelets. In this study, we determined the complete sequence of triflavin, which is composed of a single polypeptide chain of 70 amino acids. Its sequence is rich in cysteine and contains Arg-Gly-Asp at residues 49-51 in the carboxy-terminal domain. Triflavin shows about 68% identity of amino acid sequence with trigramin, which is a specific antagonist of the fibrinogen receptor associated with glycoprotein IIb/IIIa complex. [125I]Triflavin binds to unstimulated and ADP-stimulated platelets in a saturable manner and its Kd values are estimated to be 76 and 74 nM, respectively; the corresponding numbers of binding sites are 31,029 and 34,863 per platelet, respectively. [125I]Triflavin binding is blocked by Gly-Arg-Gly-Asp-Ser in a competitive manner. EDTA, the Arg-Gly-Asp-containing peptides (including naturally occurring polypeptides, trigramin and rhodostomin), and monoclonal antibody, 7E3, raised against GP IIb/IIIa complex, inhibit [125I]triflavin binding to unstimulated and ADP-stimulated human platelets. In conclusion, triflavin specifically binds to fibrinogen receptor associated with GP IIb/IIIa complex and its binding site is located at or near GP IIb/IIIa complex, overlapping with those of 7E3 and another Arg-Gly-Asp-containing polypeptide, rhodostomin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

[Gas chromatographic/mass spectrometric analysis of boldenone urinary metabolites in man].

The metabolism of boldenone (17 beta-hydroxy-1,4-androstem-3-one) in man has been investigated by gas chromatography/mass spectrometry. After oral administration of a 20 mg dose to man, six metabolites were detected in the conjugated fraction of the urinary samples. Boldenone, the major compound excreted in urine, was detected within 34 h after administration. In addition, several metabolites, resulting from the hydroxylation of boldenone and the reduction of the unsaturated carbon bonds of boldenone, were detected in the urine samples varying from 9 to 83 h. Extraction and fractionation of these metabolites were achieved by using XAD-2 column and gas chromatography. The recovery of the whole procedure was studied. Furthermore, the mass spectra of the metabolites are presented and major fragment pathways are discussed.

Gas Chromatography-Mass Spectrometry↗

Purification, characterization, and molecular cloning of gonadotropin subunits of silver carp (Hypophthalmichthys molitrix).

The alpha and beta subunit of silver carp gonadotropin (scGTH-alpha and scGTH-beta) were isolated by high-performance liquid chromatography. Heterogeneity of N-terminal amino acid sequence was observed in scGTH-alpha but not in scGTH-beta. For determining the complete primary structures of scGTH-alpha and scGTH-beta, their cDNAs were cloned. Combining the data of N- and C-terminal sequences determined from proteins and the amino acid sequences deduced from cDNAs, we infer that scGTH-alpha consists of 95 and/or 93 residues and scGTH-beta consists of 115 residues. Both scGTH-alpha and scGTH-beta are glycoprotein. Their carbohydrate content is about 20 g per 100 g protein. The molecular weights of scGTH-alpha and scGTH-beta were calculated to be 12,700 and 15,700 Da, respectively. The amino acid sequences of scGTH-alpha and scGTH-beta are very similar to those of the corresponding subunit of carp GTH, different in only 2 and 4 residues, respectively. In addition, a high extent of homology (70%) was also observed between the alpha subunits of fish and mammalian GTHs. In the case of beta subunit, homology among various species of fish (75 to 98%) is much higher than that between fish and mammal (40%). These data suggest that the alpha subunit is conserved while the beta subunit is diversified during the molecular evolution of vertebrate GTH.

Amino Acid Sequence↗

Revised amino acid sequences of the three major phospholipases A2 from Bungarus fasciatus (banded krait) venom.

The structures of three cardiotoxin-like proteins obtained from the venom of Bungarus fasciatus (banded krait) were elucidated previously (Lu and Lo, 1980, 1981). Since their molecular sizes are similar to that of phospholipase A2 and since they show weak phospholipase A2 activities (Chang et al., 1983), a further study of their primary structures was carried out. Fractions Va, Vb-2 and VI, corresponding to the former V-2, V-3 and VI were determined to be typical phospholipases A2. Among 118 amino acid residues, they all have in common a Pro29 between Gly28 and Gly30, the latter two residues being implicated in Ca2+ binding together with Tyr26 and Asp47.

Amino Acid Sequence↗

Pike eel (Muraenesox cinereus) gonadotropin. Amino acid sequences of both alpha and beta subunits.

The amino acid sequences of pike eel gonadotropin alpha and beta subunits have been determined by standard sequencing analytical methods. The alpha subunit is composed of 93 amino acid residues while the beta subunit comprises 113 amino acid residues. All the invariant half-cystine residues are in the same positions as those found in other gonadotropins. It is noteworthy that the first, putative glycosylation site (Asn56) found in the alpha subunit of other gonadotropins was replaced by Asp56 in the alpha subunit of pike eel gonadotropin. Similarity analyses indicate that both subunits are structurally more similar to other known fish gonadotropin subunits than to those of the mammalian gonadotropins.

Amino Acid Sequence↗

Unusual amino acid sequence of fasciatoxin, a weak reversibly acting neurotoxin in the venom of the banded krait, Bungarus fasciatus.

A weak reversibly acting neurotoxin, fasciatoxin, was found in the venom of Bungarus fasciatus. The sequencing was completed by manual and automated Edman analyses of the reduced and carboxymethylated protein and of the peptides obtained from enzyme digestions. It is composed of 63 amino acid residues with four disulphide bonds and a unique sequence at the C-terminal end. According to the criteria set by Ryden, Gabel & Eaker [(1973) Int. J. Pept. Protein Res. 5, 261-273], fasciatoxin lacks all of the five functionally invariant residues of neurotoxins. The hydropathy index indicates that fasciatoxin is devoid of a strong hydrophilicity domain for binding to the receptor site. Structural comparison with some typical neurotoxins also reveals the uniqueness of fasciatoxin in that the extent of similarity is only about 30%.

Amino Acid Sequence↗

Amino acid sequence of a short chain neurotoxin from the venom of banded krait (Bungarus fasciatus).

The amino acid sequence of a short chain neurotoxin obtained from Bungarus fasciatus venom consists of 64 amino acid residues: Arg-Ile-Cys-Leu-Asn-Gln-Gln-Gln-Ser- Thr-Pro-Glu-Asp-Gln-Pro-Thr-Asn-Gly-Gln-Cys-Tyr-Ile-Lys-Thr-Asp-Cys-Gln- Asn-Lys - Thr-Trp-Asn-Thr-His-Arg-Gly-Ser-Arg-Thr-Asp-Arg-Gly-Cys-Gly-Cys-Pro-Lys- Val-Lys - Pro-Gly-Ile-Asn-Leu-Arg-Cys-Cys-Lys-Thr-Asp-Lys-Cys-Asn-Glu. The above result was obtained primarily from the amino acid analyses and sequencing of tryptic peptides accompanied with the necessary analyses and sequencing of the chymotryptic and lysyl endopeptidic peptides for alignment.

Amino Acid Sequence↗

Amino acid sequence of a neutral phospholipase A2 (III) in the venom of Bungarus fasciatus.

Two phospholipases were found in the venom of Bungarus fasciatus, one in fraction III, the other in fraction X of the chromatographic separation. A neutral PLA2(III) purified from fraction III was subjected to amino acid sequencing by means of an automated sequenator applied to the intact RCM-PLA2 (III) and the individual peptides obtained from HPLC separation of the three types of enzymatic peptides. PLA(III) was shown to consist of 118 amino acid residues with 14 half-cystines. It is 65% homologous to the basic PLA2 obtained from fraction X.

Amino Acid Sequence↗

Double-blind crossover controlled study on antiepilepsirine.

Antiepilepsirine (AES) is one of the derivatives of a Chinese folk prescription. It is a new antiepileptic drug (AED) synthesized in cooperation by Chinese medical and pharmaceutical workers. Pharmacological experiments on animal models prove that its antiepileptic action is marked, but there has been little evaluation of its clinical effects. We used the double-blind placebo-controlled crossover study which is generally acceptable as a proper method for new drug study. This study covered 58 epileptic children treated with classical AEDs and observed 6.5 months. Every patient took AES and placebo for 3 months each by random crossover on the basis of add-on therapy. The blood levels of AES and other antiepileptic drugs were determined regularly. The results show that there are no significant differences in clinical effects between AES and placebo in pediatric epilepsies as a whole, but AES is effective in tonic-clonic seizures (P less than 0.05), the most common type of seizure in the series. There are no significant differences in AED blood levels between the AES effective and ineffective groups. AES has no effect on the blood levels of other AEDs. AES is very safe, children given large doses (10 mg/kg/day) demonstrate no serious side-effects. It is suggested that there is potential improvement in patient psychological and cognitive status. This article also discusses the evaluation of new drugs for clinical effects, subject sampling, the criteria for efficacy evaluation and relationship between animal information and human outcome. The AES chemical structure is different from other well-known AEDs, this is a unique advantage.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗