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Biomedical subjects

A Q Wang

Publications and source records attributed to A Q Wang.

14 recordsLinked to original sources

High-throughput sample preparation procedures for the quantitation of a new bone integrin alpha(nu)beta(3) antagonist in human plasma and urine using liquid chromatography-tandem mass spectrometry.

High throughput LC-MS/MS assays to quantitate a new alpha(nu)beta(3) bone integrin antagonist (I) in human plasma and urine have been developed using instruments programmed to automate sample preparation procedures. Packard liquid handling system-MultiPROBE II EX was programmed for preparing calibration standards in control plasma and urine, acidifying all standards, quality control (QC), and clinical samples with necessary dilutions, and adding the internal standard to the acidified samples. TOMTEC Quadra 96 was programmed to perform the solid phase extraction (SPE) process on a 3M 96-well mixed phase cation standard density (MPC-SD) plate to isolate the analytes from the sample matrix. The extract collected from both types of matrices was directly injected into reversed-phase LC-MS/MS system with a Turbo Ion Spray (TIS) interface in the positive ionization mode. The plasma and urine assays have the calibration range of 0.5-1500 and 2-6000 ng/mL, respectively. Validation of the automated and the manual plasma assays showed that application of MultiPROBE II to sample preparation gave comparable accuracy and precision. Overall, the automated approaches with minimum manual intervention enhanced the throughput of sample preparation.

Bone and Bones↗

Determination of a beta(3)-agonist in human plasma by LC/MS/MS with semi-automated 48-well diatomaceous earth plate.

Methods for the determination of a beta(3)-agonist (A) in human plasma were developed and compared based on high-performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection using a turbo ion spray (TIS) interface. Drug and internal standard were isolated from plasma by three sample preparation methods, liquid-liquid extraction, Chem Elut cartridges and 48-well diatomaceous earth plates, that successively improved sample throughput for LC/MS/MS. MS/MS detection was performed on a PE Sciex API 365 tandem mass spectrometer operated in positive ion mode and using multiple reaction monitoring (MRM). The precursor/product ion combinations of m/z 625/607 and 653/515 were used to quantify A and internal standard, respectively, after chromatographic separation of the analytes. Using liquid-liquid extraction and Chem Elut cartridges, the assay concentration range was 0.5-100 ng/ml. Using diatomaceous earth plates, the concentration range of the assay was extended to 0.5-200 ng/ml. For all three assays, the statistics for precision and accuracy is comparable. The assay accuracy ranged from 91-107% and intraday precision as measured by the coefficient of variation (CV) ranged 2-10%. The sample throughput was tripled when the diatomaceous earth plate method was compared with the original liquid-liquid extraction method.

Adrenergic beta-3 Receptor Agonists↗

[Studies of gene regulation of de novo biosynthetic pathway of purine in Salmonella typhimurium. X. Isolation of purR(am) mutants and preliminary studies of amino acid substitution].

Starting from a super-repressed mutant of purR, 3-18, 439 independent candidates of purR- mutants were isolated by using NCE selecting plate with lactose as sole carbon source. Among these mutants. 11 amber mutants were detected by introducing a tRNA suppressor gene. Cotransduction analysis proved that the amber mutation sites of 11 amber mutants all located on purR. Amino acid substitution experiments were performed with three tRNA suppressors, supD, supE and supF, for each purR(am). The results showed that the same amino acid substitution occurred in different site of PurR protein could result in varied effects on purR function; different amino acid substitution occurred at the same position of PurR protein also could produced varied effects on purR function.

Bacterial Proteins↗

[Regulation of purine biosynthetic genes expression in Salmonella typhimurium].

To study binding funtion of 4 consensus bases in 16 bp PUR box with purR protein, the directed site mutation for each was carried out, which mutate from C to G, A to G, A to G, T to C, respectively. Gel retardation showed that the PUR box carrying a reserved mutation could not bind with purR protein. It suggested that all these consensus base pairs are necessary to hold the normal binding function of PUR box with purR protein.

Gene Expression Regulation, Bacterial↗

Diagnostic yield of repeated smear microscopy examinations among patients suspected of pulmonary TB in Shandong province of China.

OBJECTIVES: To analyse the yield of five repeated smear microscopy examinations for the diagnosis of smear-positive pulmonary tuberculosis (TB). METHODS: Patients with respiratory symptoms and abnormal chest X-rays provided five spontaneous sputum samples for acid-fast bacilli (AFB) smear microscopy in one of nine county laboratories. RESULTS: Of 9302 patients with respiratory symptoms and abnormal X-rays, 6437 (69%) had at least one smear-positive sputum. Of these, 84.5% were diagnosed on the first smear, 96.7% on the first two smears, and 99.9% on the first three sputum smears. The fourth and fifth sputum smears yielded only seven additional cases (0.1%). CONCLUSIONS: Smear microscopy examination of two spontaneous sputum specimens is the most efficient, and three sputum smear examinations provide a diagnosis in almost all cases.

China↗

[Studies on non-anthraquinones in Rheum officinale Baill].

OBJECTIVE: To investigate the non-anthraquinones from the root and rootstock of Rheum officinale. METHOD: The chemical constituents were obtained from the 85% alcohol extract of the radix and rhizoma of R. officinale by column chromatography and identified by spectroscopic analysis. RESULT: Ten non-anthraquinones were obtained, of which eight were identified by spectroscopic analysis as rheosmine, daucosterol, d-catechin, 6-cinnamoylisolindleyin, (-)-epicatechin-3-O-gallate, resveratrol-4'-O-beta-D-(6"-O-galloyl)-glucopyranoside, gallic acid, D-sorbitol. CONCLUSION: Compounds rheosmine and D-sorbitol were obtained from the genus Rheum for the first time.

Plant Roots↗

[Expression regulation of purine biosynthetic genes in Salmonella typhimurium. VI. Isolation and characterization of super-repressor mutants].

Starting from strain of Salmonella typhimurium purD::lac, 86 exponential cultures were mutagenized with NTG and white or light blue clones on E + ado + Xgal plate were selected as candidates of purRs mutant. Total 66 independent candidate strains were obtained. By assaying their beta-galactosidase activity under the repressed and derepressed conditions, determining their frequency of revertional mutation, Conducting transductional analysis of mutational site and dorminance test, 11 candidates strains were proved to be super-repressor mutants. These mutants are useful for studying the expression of purine biosynthetic gene and relationship between protein structure and function in general.

Bacterial Proteins↗

Hyperselective posterior rhizotomy in treatment of spasticity of paralytic limbs.

One hundred and eight patients with spasticity of the paralytic limbs were treated successfully with hyperselective posterior rhizotomy (SPR). Of the 108 patients, 100 had cerebral palsy, 2 hemiplegia, 3 sequelae of cerebral injury, 2 paraplegia and 1 multiple sclerosis. Twelve patients received cervical SPR and 96 lumbosacral SPR. Laminectomy is performed to open the dura and to separate the posterior spinal root into several rootlets. The lower threshold rootlets were divided after electrical stimulation. Follow-up for 6 to 30 months showed that the effective rate of reducing spasticity was over 95% and functional improvement rate over 80%.

Adolescent↗

[Regulation of riboflavin biosynthesis in Salmonella typhimurium].

7 independent rib genes fusions with MudJ (lacZ, Kanr) were isolated by transposon MudJ mutagenesis in Salmonella typhimurium. 5 of them are blue on the X-gal plate, and the beta-galactosidase activity of the cells grown in E medium containing various concentration of riboflavin were assayed. The results showed that the expression of rib gene are not repressed by riboflavin. It appears to be synthesized constitutively in Salmonella typhimurium.

Gene Expression Regulation, Bacterial↗

Phosphate groups as substrate determinants for casein kinase I action.

Phosphorylation of rabbit muscle glycogen synthase by cyclic AMP-dependent protein kinase has been shown to enhance subsequent phosphorylation by casein kinase I (Flotow, H., and Roach, P. J. (1989) J. Biol. Chem. 264, 9126-9128). In the present study, synthetic peptides based on the sequences of the four phosphorylated regions in muscle glycogen synthase were used to probe the role of substrate phosphorylation in casein kinase I action. With all four peptides, prior phosphorylation significantly stimulated phosphorylation by casein kinase I. A series of peptides was synthesized based on the NH2-terminal glycogen synthase sequence PLSRTLS7VSS10LPGL, in which phosphorylation at Ser7 is required for modification of Ser10 by casein kinase I. The spacing between the P-Ser and the acceptor Ser was varied to have 1, 2, or 3 intervening residues. The peptide with a 2-residue spacing (-S(P)-X-X-S-) was by far the best casein kinase I substrate. When the P-Ser residue at Ser7 was replaced with P-Thr, the resulting peptide was still a casein kinase I substrate. However, substitution of Asp or Glu residues at Ser7 led to peptides that were not phosphorylated by casein kinase I. Phosphorylation of one of the other peptides showed that Thr could also be the phosphate acceptor. From these results, we propose that there are substrates for casein kinase I for which prior phosphorylation is a critical determinant of protein kinase action. In these instances, an important recognition motif for casein kinase I appears to be -S(P)/T(P)-Xn-S/T- with n = 2 much more effective than n = 1 or n = 3. Thus, casein kinase I may be involved in hierarchal substrate phosphorylation schemes in which its activity is controlled by the phosphorylation state of its substrates.

Amino Acid Sequence↗

In vivo cloning of proline genes and its expression in Escherichia coli.

The wild type proA+, B+ genes of E. coli were cloned in vivo using a plasmid containing a mini-Mu replicon, pEG5005. The cloning frequency was about 1.46 x 10(-3)/Kanr transductant. Genetic and biochemical analysis of these clones indicated that the proA+, B+ genes are on the plasmid pEG5005. The secretion of proline were assayed for 500 Pro+ clones. However, no proline accumulation was detected. A Pro+ clone pPR3 was mutagenized in vivo by NTG and the mutants resistant to D-proline were obtained. One of the Dpr mutants pPR7 was found to produce 0.35 mg/ml proline in a proA B deletion strain. When pPR7 was transferred into a proline producing strain, the yield of proline increased up to 2.5 mg/ml, which is 7 and 2.5 times higher than that of the donor and recipient respectively. The physical maps of pEG5005 and pPR3 were roughly established.

Chromosome Mapping↗

[Transpon Tn5 mutagenesis in Citrobacter].

When E. coli 1830/PJB4JI mating with four Citrobacter strains all were Kanamycin resistant, but a majority of KanrGens transconjugants were obtained from C-3-1. Among 3000 KanrGens 21 were auxotrophs, these are Lys-(1), Ura-(1), Arg-(2), Iso-(2), His-(2), Met-(1), Phe-(1), Tyr-(1), Ser-(1), Thr-(1), Leu-(3), Pro-(1), Ade-(3), Lac-(1), PJB4JI plasmid DNA were detected in parent strain E. coli 1830, but not in auxotrophs strains which carrying Tn5 induced mutations. Twenty auxotrophs chromosome DNA were hybridized with Tn5 DNA labeled with 32P respectively, all auxotrophs has positive reaction. Therefore, we concluded from genetic and physical data that auxotrophs resulted from Tn5 transposition from PJB4JI into C-3-1 chromosome.

Citrobacter↗