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Q Dong

Publications and source records attributed to Q Dong.

156 records · Page 9Linked to original sources
PubMed1990

Identification of different classes of nonessential sulfhydryl groups in Escherichia coli adenylosuccinate synthetase.

Reaction of Escherichia coli adenylosuccinate synthetase with the thiol reagent 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) or N-ethylmaleimide (NEM) leads to modification of one cysteine residue per enzyme monomer without significant loss of enzyme activity. Modification of a second cysteine residue occurs under mild denaturing conditions (3.5 M urea), and derivatization of this thiol followed by dialysis results in complete loss of enzyme activity. The remaining two cysteine residues react with DTNB only after treatment with 8 M urea. The location of the various cysteine residues in the enzyme was established by using [14C]NEM followed by tryptic digestion and radiopeptide isolation. The reactive cysteine has been identified as Cys291, and the thiol exposed with 3.5 M urea is Cys344. When Cys344 was replaced by either serine or alanine, the mutant enzymes were found to be as active as the wild-type enzyme. These findings point to the nonessential role of Cys344 in adenylosuccinate synthetase.

Adenylosuccinate Synthase↗
PubMed1990

Comparison of O-6-methylguanine DNA methyltransferase (MGMT) mRNA levels in Mer+ and Mer- human tumor cell lines containing the MGMT gene by the polymerase chain reaction technique.

O-6-Alkylguanine is a mutagenic and carcinogenic DNA lesion induced by a variety of alkylating agents, including the chloroethylnitrosoureas. The lesion is repaired by the alkyl-accepting suicide enzyme O-6-methylguanine DNA methyltransferase (MGMT). Approximately 25% of cell lines derived from human tumors are phenotypically deficient in this enzyme and are described as Mer-. Recent cloning of the human MGMT cDNA (Tano, K.; Shiota, S.; Collier, J.; Foote, R.S.; Mitra, S. Proc. Natl. Acad. Sci. USA 87:686-690; 1990) has allowed for a more detailed analysis of the basis of the Mer- phenotype in human Mer- tumor cell lines. Using the polymerase chain reaction (PCR) technique, an MGMT cDNA probe based on the published sequence was generated. The probe and the PCR technique were then used to analyze the presence and expression of the MGMT gene in two Mer+ and four Mer- lines, including one SV40-transformed Mer- line and three Mer- human tumor cell lines. The data demonstrate that while all six cell lines contained a relatively nonamplified, nonrearranged MGMT gene, Mer- lines contained levels of MGMT mRNA detectable only by PCR analysis. Of the three Mer- tumor cell lines examined, two (COLO 320 HSR, A1235) contained MGMT mRNA levels that were four to five orders of magnitude lower than that of the prototype Mer+ tumor line (HT-29), while one (BE) contained no consistently detectable MGMT mRNA. These results suggest that in the human Mer- tumor lines tested, the Mer- phenotype was mediated by a severe reduction in MGMT mRNA levels, despite the presence of the MGMT gene.

Base Sequence↗
PubMed1990

[An acute experimental study on the combinations of aluminum and fluorine in various ratios].

The joint actions were assessed by four methods after male Wistar rats had been acutely intoxicated with nine proportional mixtures of Al and F. Strong antagonistic effects of Al on F were observed in the mixtures, in which the ratio of Al to F ranged from 0.8:0.2 to 0.1:0.9. The optimum ratio of Al to F for antagonism ranged from 0.3:0.7 to 0.1:0.9 without periorbital bleeding that was a regular finding in Al acute toxicity on the rat to be seen. When the amount of Al in the mixture was below 5% or above 90%, no antagonistic effect was shown. The kinetics on the absorption under the condition of Al and F existing simultaneously is still unknown, and warrants further investigation.

Aluminum↗
PubMed1989

Towards an antigenic map of human thyroglobulin: identification of ten epitope-bearing sequences within the primary structure of thyroglobulin.

The aim of this study was to define, at the molecular level, epitopes of thyroglobulin recognized by heterologous antibodies and autoantisera. One hundred thousand clones from a lambda gt 11 human thyroid cDNA library were screened using a rabbit antibody to human thyroglobulin at a 1:2000 dilution. Twenty clones were plaque-purified to homogeneity, and characterization and sequencing of their cDNA inserts showed that they represented four distinct regions of the thyroglobulin molecule, one of them being the 22 carboxyl-terminal amino acids. Rescreening of the library with the same rabbit antibody to human thyroglobulin absorbed with peptides encoded by the carboxyl-terminal clone, led to the definition of six further epitope-bearing fragments of thyroglobulin. The ten regions that we have identified were recognized by ten further rabbit antibodies to human thyroglobulin, showing that they are representative of the repertoire of heterologous epitopes. In contrast, none of the ten heteroepitope-bearing fragments was recognized by sera from ten patients with autoimmune thyroid disease with various titres of thyroglobulin antibodies. Screening of 2 x 10(6) clones from the library using a pool of ten autoantisera (individual sera diluted to 1:1000), and of 1 x 10(6) clones using a single autoantiserum of very high antithyroglobulin titre (diluted 1:400) resulted in no thyroglobulin clones being isolated. The significance of these results to the immune process is discussed.

Amino Acid Sequence↗
PubMed1989

Definition, at the molecular level, of a thyroglobulin-acetylcholinesterase shared epitope: study of its pathophysiological significance in patients with Graves' ophthalmopathy.

The nature of the putative autoantigen in Graves' ophthalmopathy (Go) remains an enigma but the sequence similarity between thyroglobulin (Tg) and acetylcholinesterase (ACHE) provides a rationale for epitopes which are common to the thyroid gland and the eye orbit. In an attempt to define the shared epitope, we have screened a lambda gt 11 human thyroid cDNA library using a polyclonal antibody to Torpedo ACHE and isolated two clones, which upon sequencing, were shown to contain Tg segments, corresponding to portions of the C terminal part of the molecule which has a high similarity with ACHE. Having demonstrated the existence of an epitope common to Tg and ACHE, the clones have been further tested and found to be positive in lysis plaque assays with 1/10 sera from patients with Hashimoto's thyroiditis (HT), 8/8 from patients with Graves' ophthalmopathy and 0/8 normal sera. We have investigated the physiological significance of this common epitope by in situ immunolocalization studies in which the polyclonal antibody to Torpedo ACHE (which was used for screening the library) and immunoglobulins (Igs) from 6 Go patients tested were shown to bind to end plate regions of human foetal muscle fibres which were concurrently shown to be rich in cholinesterase activity: Igs from 3 normal individuals and 2 patients with Hashimoto's thyroiditis did not bind. The results demonstrate and characterize an epitope which is common to Tg and ACHE and show that Go patients Igs contain antibodies which bind to muscle end plates rich in cholinesterase. The significance of these findings to the pathogenesis of Go is discussed.

Acetylcholinesterase↗
PubMed1988

Cellular and extracellular plasminogen activator and inhibitor in an experimental tumour.

We determined plasminogen activator (PA) and PA inhibitor (PAI) activities in the intra- and extracellular compartments of an experimental pancreatic ascites tumour with indirect and direct functional assays, and partially characterized these activities on SDS-polyacrylamide gels coupled with fibrin and reverse fibrin autography. Intact tumour cells caused lysis of plasminogen-rich but not plasminogen-free fibrin clots, and the extent of lysis of the former was related to tumour cell count. Direct assay of PA with a synthetic substrate yielded an equivalent of 109 urokinase units per 10(9) tumour cells. No PAI activity was demonstrated in tumour cells with functional assays. Contrary to tumour cells, cell-free ascitic fluids caused no lysis of fibrin clots. Instead, it inhibited tumour cell- and urokinase-induced, but not plasmin-induced, clot lysis in a dose-dependent fashion. Although functional assays failed to demonstrate PA in ascitic fluid and PAI in tumour cells, both activities were detected in electrophoresed samples of cell lysates and fluids by fibrin and reverse fibrin autography. In tumour cells, a mixture of tissue-type PA (tPA) and urokinase-type PA (uPA) were present. In the fluid, uPA together with two other PAs with greater molecular weights than tPA were detected.

Animals↗
PubMed1988

Susceptibility of irradiated bovine aortic endothelial cells to injury.

Using cultured bovine aortic endothelial cells (BAEC), the authors attempted to determine whether prior irradiation would alter the susceptibility of these cells to three known injurious stimuli and, if so, whether the alteration would be related to radiation dose. BAEC were irradiated with 0, 5, or 10 Gy of gamma rays and, on the third postirradiation day, exposed to fibrin, nicotine, or bacterial endotoxin (lipopolysaccharide, LPS). Release of prelabeled 51Cr, representing cell lysis, cell detachment, or a combination of the two, was determined. Significant differences between irradiated and control cells were determined by using paired Student's t-tests. Irradiation did not appear to have altered the sensitivity of BAEC to fibrin-induced injury. Cells irradiated with 10 Gy of gamma rays, but generally not those irradiated with half this dose, showed a heightened susceptibility to nicotine. Contrary to the nicotine results, irradiated cells showed less cell detachment and lysis after exposure to LPS. These results suggest that the susceptibility of irradiated BAEC to harmful stimuli depends largely on the nature of the stimulus as well as the radiation dose.

Animals↗
PubMed1987

A nonsense mutation causes hereditary goitre in the Afrikander cattle and unmasks alternative splicing of thyroglobulin transcripts.

The hereditary goitre of Afrikander cattle is an autosomal recessive disease characterized in homozygotes by the production of abnormal thyroglobulin (Tg) and the coexistence in the thyroid of normal-sized 8.4-kilobase (kb) Tg mRNA with a misspliced 7.3-kb message having lost exon 9. We have cloned and sequenced the cDNA segment corresponding to the abnormal exon 8-exon 10 junction and the relevant genomic DNA region. The mutation responsible for the disease is a cytosine to thymine transition creating a stop codon at position 697 in exon 9. The original reading frame is maintained in the 7.3-kb mRNA, which, as it lacks the mutated exon, is translatable into a potentially functional protein. This puzzling phenotype in which a mutated exon is apparently removed selectively from transcripts by alternative splicing leads us to suggest that the 7.3-kb transcript could be present in normal animals. Using a sensitive oligonucleotide hybridization assay, we have demonstrated that a 7.3-kb mRNA lacking exon 9 does exist in normal thyroids as a minor mRNA species. As it is fully translatable, the 7.3-kb mRNA is expected to be more stable than the normal-sized 8.4-kb message. This probably accounts for the higher proportion of 7.3-kb transcript found in the goitre.

Amino Acid Sequence↗
PubMedDate not supplied

New global communication process in thermodynamics: impact on quality of published experimental data.

Thermodynamic data are a key resource in the search for new relationships between properties of chemical systems that constitutes the basis of the scientific discovery process. In addition, thermodynamic information is critical for development and improvement of all chemical process technologies. Historically, peer-reviewed journals are the major source of this information obtained by experimental measurement or prediction. Technological advances in measurement science have propelled enormous growth in the scale of published thermodynamic data (almost doubling every 10 years). This expansion has created new challenges in data validation at all stages of the data delivery process. Despite the peer-review process, problems in data validation have led, in many instances, to publication of data that are grossly erroneous and, at times, inconsistent with the fundamental laws of nature. This article describes a new global data communication process in thermodynamics and its impact in addressing these challenges as well as in streamlining the delivery of the thermodynamic data from "data producers" to "data users". We believe that the prolific growth of scientific data in numerous and diverse fields outside thermodynamics, together with the demonstrated effectiveness and versatility of the process described in this article, will foster development of such processes in other scientific fields.

Chemistry↗
PubMedDate not supplied

Diagnosis of renal vascular disease with MR angiography.

Renal magnetic resonance (MR) angiography allows accurate evaluation of patients suspected to have renal artery stenosis without the risks associated with nephrotoxic contrast agents, ionizing radiation, or arterial catheterization. Other applications of renal MR angiography are mapping the vascular anatomy for planning renal revascularization, planning repair of abdominal aortic aneurysms, assessing renal bypass grafts and renal transplant anastomoses, and evaluating vascular involvement by renal tumors. A variety of pulse sequences provide complementary information about kidney morphology, arterial anatomy, blood flow, and renal function and excretion. Three-dimensional gadolinium-enhanced MR angiography can be combined with several other sequences to produce a comprehensive approach to renal MR angiography. This comprehensive approach is designed to allow hemodynamic characterization of renal artery stenosis with a single MR imaging examination that can be easily completed in 1 hour. Three-dimensional gadolinium-enhanced MR angiography demonstrates the renal arteries along with the abdominal aorta, iliac arteries, and mesenteric arteries in a 20-30-second acquisition that can be performed during breath holding. Numerous projections are reconstructed from a single three-dimensional volume of data acquired with a single injection of contrast material to obtain perpendicular and optimized views of each renal artery.

Anastomosis, Surgical↗
PubMedDate not supplied

MR angiography after renal revascularization: spectrum of expected anatomic results and postintervention complications.

The use of magnetic resonance (MR) angiography in screening for renal artery stenosis has been extensively evaluated. However, the MR angiographic findings after renal artery revascularization are not as well characterized. The renal artery and parenchyma can be evaluated after revascularization with a comprehensive MR imaging protocol that includes T1- and T2-weighted spin-echo sequences, three-dimensional (3D) gadolinium-enhanced MR angiography, and 3D phase-contrast MR angiography. Because surgical techniques for revascularization vary, knowledge of the surgical procedure is necessary to ensure inclusion of the pertinent anatomy at 3D gadolinium-enhanced MR angiography and to define appropriate 3D phase-contrast MR angiography volumes. The 3D gadolinium-enhanced MR angiography volume can be manipulated to view relevant vascular anatomy at the optimal obliquity and section thickness. This protocol allows robust, noninvasive evaluation of the expected arterial anatomy after revascularization, including renal artery endarterectomy, aortorenal bypass grafts, and extraanatomic reconstructions. In cases of suspected postrevascularization complications, gadolinium-enhanced MR angiography is useful because of its lack of nephrotoxicity and radiation exposure. Immediate complications of renal revascularization include renal artery thrombosis, renal infarction, and perinephric hemorrhage. Long-term complications include aneurysms of bypass grafts and recurrent stenosis of the renal artery.

Adult↗