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Biomedical subjects

Y G Tsay

Publications and source records attributed to Y G Tsay.

14 recordsLinked to original sources
PubMed2003

Management of aortic aneurysm infected with Salmonella.

BACKGROUND: This study reviewed the clinical outcomes of patients with an aortic aneurysm infected with Salmonella treated by a single centre over 6 years. METHODS: Data were collected by a retrospective case-note review. RESULTS: Between September 1995 and December 2001, 121 patients with non-typhoid Salmonella bacteraemia were treated, of whom 24 patients had an aortic aneurysm infected with Salmonella. Ten had a suprarenal and 14 an infrarenal aortic infection. The most common responsible pathogen was group C Salmonella (12 patients). All of the 20 patients who had combined medical and surgical therapy survived, whereas two of four who had medical therapy alone died. There were two late deaths during a mean follow-up of 23 (range 3-63) months. CONCLUSION: The incidence of aortic infection in patients with non-typhoid Salmonella bacteraemia was high in Taiwan. Timely surgical intervention and prolonged intravenous antibiotic therapy resulted in excellent outcomes.

Abdominal Pain↗
PubMed2001

A 60-kilodalton immunodominant glycoprotein is essential for cell wall integrity and the maintenance of cell shape in Streptococcus mutans.

We have demonstrated previously by Western blotting that in naturally sensitized humans, the serum or salivary antibody response to Streptococcus mutans was directed predominantly to a protein antigen with a size of approximately 60-kDa. To identify this immunodominant antigen, specific serum antibodies were eluted from immunoblots and five positive clones with inserts ranging in length from 3 to 8 kb from identical chromosomal loci were obtained by screening a genomic expression library of Streptococcus mutans GS-5. Amino acid sequencing established the identity of this immunodominant antigen, a 60-kDa immunodominant glycoprotein (IDG-60), to be a cell wall-associated general stress protein GSP-781, which was originally predicted to have a molecular mass of approximately 45 kDa based on the derived nucleotide sequence. Discrepancy in the molecular mass was also observed in recombinant his-tagged IDG-60 (rIDG-60) expressed from Escherichia coli. Glycosylation, consisting of sialic acid, mannose galactose, and N-acetylgalactosamine, was detected by lectin binding to IDG-60 in cell wall extracts from S. mutans and rIDG-60 expressed in vivo or translated in vitro. Despite the presence of multiple Asn or Ser or Thr glycosylation sites, IDG-60 was resistant to the effect of N-glycosidase F and multiple O-glycosidase molecules but not to beta-galactosidase. Insertional inactivation of the gene encoding IDG-60, sagA, resulted in a retarded growth rate, destabilization of the cell wall, and pleiomorphic cell shape with multifold ingrowth of cell wall. In addition, distinct from the parental GS-5 strain, the isogenic mutant GS-51 was unable to survive the challenge of low pH and high osmotic pressure or high temperature. Expression of the wild-type gene in trans within GS-51 from plasmid pDL277 complemented the growth defect and restored normal cell shape. These results suggested that IDG-60 is essential for maintaining the integrity of the cell wall and the uniformity of cell shape, both of which are indispensable for bacteria survival under stress conditions.

Adult↗
PubMed2000

A strategy for identification and quantitation of phosphopeptides by liquid chromatography/tandem mass spectrometry.

Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is a state-of-the-art method of structural analysis of peptides/proteins. Here, using activating transcription factor-2 (ATF2) as an example, we report how LC/MS/MS data were processed to generate selected ion tracings for identification of phosphorylated peptides based on their parallel elution behavior with their nonphosphorylated analogs. Via this approach, we verified that amino acid residues Thr-69, Thr-71, and Ser-90 of ATF2 were the in vitro targets for c-Jun kinase. Selected ion tracing method was also used to quantitatively determine phosphorylation states of peptides. We demonstrated that the phosphorylation of Thr-69/Thr-71 was increased in response to ultraviolet irradiation specifically in subconfluent but not in confluent cultures. About 24% of Thr-69/Thr-71-containing segment were singly phosphorylated in subconfluent cultures, while minimal phosphorylation occurred in confluent cultures. In contrast, Ser-112 phosphorylation remained unaffected by cell densities. This strategy could be applied to the studies of a variety of modifications seen in various regulated cellular processes.

Activating Transcription Factor 2↗
PubMed1999

Export of galectin-3 from nuclei of digitonin-permeabilized mouse 3T3 fibroblasts.

Galectin-3 is a galactose-/lactose-binding protein (M(r) approximately 30,000), identified as a required factor in the splicing of pre-mRNA. Immunofluorescence staining revealed that galectin-3 distributes differentially between the nucleus and the cytoplasm, depending on the proliferative state of the cells under analysis. Using digitonin-permeabilized mouse 3T3 fibroblasts, we provide evidence that galectin-3 is rapidly and selectively exported from the nucleus. Although both phosphorylated and nonphosphorylated isoforms of galectin-3 are found in the nuclear fraction, only phosphorylated galectin-3 is identified in the exported fraction, implying that phosphorylation is important for the nuclear export of the protein. The rate of galectin-3 export is temperature dependent and is decreased by the addition of wheat germ agglutinin. More strikingly, galectin-3 export can be inhibited by the addition of leptomycin B, a drug that disrupts the interaction between the leucine-rich nuclear export signal and its receptor, CRM1 (chromosome maintenance region 1). Indeed, a putative leucine-rich nuclear export signal can be found in residues 241-249 of the murine galectin-3 sequence. Finally, gel filtration of the exported material showed that galectin-3 can be found in at least two high molecular weight complexes (approximately 650 and approximately 60 kDa), both of which can be disrupted by lactose.

3T3 Cells↗
PubMed1994

Galectin-3: differential accumulation of distinct mRNAs in serum-stimulated mouse 3T3 fibroblasts.

The murine Galectin-3 gene spans approximately 12 kb of DNA and contains six exons, with the translation initiation codon located in exon II. On the basis of restriction mapping and sequence analysis of the DNA upstream of exon II, primer extension assays, rapid amplification of cDNA ends, and ribonuclease protection assays were designed and carried out to determine the initiation site of transcription and the sequence of exon I. The results revealed at least two transcription initiation sites (alpha and delta), each of which appears to be specifically associated with the use of alternative donor splice sites, resulting in distinct mRNA species. Type I message initiates at transcription start site delta, uses splice donor site No. 2, retaining a 27 bp sequence, whereas type II message initiates at transcription start site alpha, uses splice donor site No. 1, resulting in the loss of the 27 bp sequence. Primer extension assays carried out with mRNA isolated from 3T3 fibroblasts at various times after serum stimulation indicate that while the type II message varies in level only a little over the first 20 h, there is dramatic accumulation of the type I message, which peaks at 16 h post mitogen addition.

3T3 Cells↗
PubMed1991

Optical biosensor assay (OBA).

We describe a new biosensor immunoassay involving optical diffraction to detect clinically important analytes in human body fluids. A silicon wafer is used as a support for immobilization of antigen or antibody. The protein-coated surface is illuminated through a photo mask to create distinct periodic areas of active and inactive protein. When the surface is incubated with a positive sample, antigen-antibody binding occurs only on the active areas. Upon illumination with a light source such as a laser, the resulting biological diffraction grating diffracts the light. A negative sample does not result in diffraction because no antigen-antibody binding occurs to create the diffraction grating. The presence or absence of a diffraction signal differentiates between positive and negative samples, and the intensity of the signal provides a quantitative measure of the analyte concentration. The technique is demonstrated with a quantitative assay of choriogonadotropin in serum.

Biosensing Techniques↗
PubMed1986

IgE FAST inhibition: a rapid technique for the standardization of pollen allergen extract potency.

An IgE FAST(TM) inhibition protocol has been developed as a rapid in vitro method for determining relative potencies of in vivo allergenic extracts. Results are obtained within one day and correlate well with those of the RAST inhibition protocol (r = .96), making the FAST assay a preferable procedure for inprocess potency determination during the manufacture of standardized allergenic extracts.

Allergens↗
PubMed1985

Correlation of allergy test results obtained by IgE FAST, RAST, and prick-puncture methods.

Prick skin testing and IgE FASTTM results were compared in eight atopic patients, using a comparable class system. Identical results were found in 53.5% of the cases; a total of 85.7% of the results were within one class difference. Sera from 436 patients were assayed for specific IgE to 94 allergens for a total of 436 test comparisons using IgE FASTTM and modified RAST. Identical results were found in 41.7% of cases; a total of 81.4% of the tests differed by one class or less. The IgE FAST showed good correlation both with prick testing and modified RAST. IgE FAST appears to be a valid non-radioactive, time-efficient method for detecting specific IgE to common inhalant and environmental allergens.

Allergens↗
PubMed1984

A preoperative chymopapain sensitivity test for chemonucleolysis candidates.

A fluorescence enzyme immunoassay (ChymoFAST test) for the quantitation of chymopapain-specific IgE antibody concentration in human serum is described. The IgE antibody is recognized as the major immunologic mediator for anaphylactic reactions. Serum chymopapain-specific IgE levels serve as an objective screening method for identifying patients most likely to tolerate chymopapain, thereby minimizing associated risks. Preoperative sera obtained from 11,658 chemonucleolysis candidates were tested, of which 0.94% (110/11,658) were found to be positive. Good predictive values for both positive and negative findings are evident based on 4776 postmarketing surveillances obtained from the surgeons.

Antibody Specificity↗
PubMed1984

Chymopapain-induced hypersensitivity following chemonucleolysis.

This study describes the changes in chymopapain-specific IgE antibody levels in patients following chemonucleolysis with Chymodiactin. Using the ChymoFAST method, chymopapain-specific IgE values were studied in 91 patients prior to and for 2 months post-Chymodiactin chemonucleolysis. A total of 8.8% (17/91) developed IgE levels greater than or equal to 0.06 IU/ml. Those patients with detectable IgE levels prior to chemonucleolysis were more likely than those with nondetectable preinjection levels (36.4% versus 4%) to develop chymopapain-specific IgE levels greater than or equal to 0.06 IU/ml.

Antibody Specificity↗
PubMed1981

A solid-phase fluoroimmunoassay of tobramycin.

A solid-phase fluoroimmunoassay has been developed for the determination of tobramycin in serum. The principle of this method is based on direct competition of a fluorescein-labeled and unlabeled hapten for a fixed amount of specific antibody immobilized on a dipstick sampler. The bound and free labeled hapten are separated simply by removing the sampler from the reaction mixture. The fluorescence signal from the bound fluorescein-labeled hapten is measured with a FIAX fluorometer. The fluorescein-labeled tobramycin, which is easily prepared, has excellent shelf life. The assay is fast and simple, involves only one pipetting step, and avoids the use of radiochemicals. Within-assay coefficients of variation are less than 10%. The assay correlates well with radioimmunoassay (r = 0.97) and radioenzymatic assay (r = 0.96).

Anti-Bacterial Agents↗