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Biomedical subjects

S S Yang

Publications and source records attributed to S S Yang.

At least 19 recordsLinked to original sources

Determination of tobacco alkaloids by gas chromatography with nitrogen-phosphorus detection.

An improved method, gas chromatography (GC) with nitrogen-phosphorus detection (NPD), has been used for determination of alkaloids in green and cured tobacco. Tobacco alkaloids of interest included nicotine, nornicotine, myosmine, anabasine, and anatabine. Tobacco samples were treated with a small quantity of aqueous ammonia solution to "loosen" tobacco tissue and to adjust pH, then extracted with solvent. The composition of the extraction solvent solution affected recoveries of the alkaloids, particularly nornicotine, and also contributed to other phenomena such as carry-over in the injection liner and "quenching" of the nitrogen-phosphorus detector. Use of a packed injection liner (e.g. with Carbowax-KOH on Chromosorb) to reduce carry-over was studied. Quenching of the nitrogen-phosphorus detector was eliminated by reducing the injection volume (i.e. increasing the split ratio), by use of a packed injection liner, and by reducing the amount of pretreatment with aqueous ammonia. A narrow bore capillary column (i.e. 0.18 mm id) was used to improve sensitivity and resolution and to increase the speed of GC analysis. An internal standard, 2,4'-dipyridyl, was used for quantitative measurement of these tobacco alkaloids.

Alkaloids↗

Optimization of headspace sampling using solid-phase microextraction for volatile components in tobacco.

Solid-phase microextraction (SPME) was evaluated as a tool for headspace sampling of tobacco samples. Several experimental parameters (e.g. sampling temperature, pH, moisture, and the type of SPME fibers) were optimized to improve sampling efficiency in two aspects; maximum adsorption and selective adsorption of volatile components onto SPME fibers. The effect of these parameters was often dominated by the physical and chemical nature (e.g. volatility, polarity) of target compounds, thus, SPME sampling conditions can be adjusted to favor a selected group of compounds, such as organic acids in tobacco.

Gas Chromatography-Mass Spectrometry↗

Effect of pH and soybean cultivars on the quantitative analyses of soybean rhizobia populations.

Quantitative analyses of fast- and slow-growing soybean rhizobia populations in soils of four different provinces of China (Hubei, Shan Dong, Henan, and Xinjiang) have been carried out using the most probable number technique (MPN). All soils contained fast- (FSR) and slow-growing (SSR) soybean rhizobia. Asiatic and American soybean cultivars grown at acid, neutral and alkaline pH were used as trapping hosts for FSR and SSR strains. The estimated total indigenous soybean-rhizobia populations of the Xinjiang and Shan Dong soil samples greatly varied with the different soybean cultivars used. The soybean cultivar and the pH at which plants were grown also showed clear effects on the FSR/SSR rations isolated from nodules. Results of competition experiments between FSR and SSR strains supported the importance of the soybean cultivar and the pH on the outcome of competition for nodulation between FSR and SSR strains. In general, nodule occupancy by FSRs significantly increased at alkaline pH. Bacterial isolates from soybean cultivar Jing Dou 19 inoculated with Xinjiang soil nodulate cultivars Heinong 33 and Williams very poorly. Plasmid and lipopolysaccharide (LPS) profiles and PCR-RAPD analyses showed that cultivar Jing Dou 19 had trapped a diversity of FSR strains. Most of the isolates from soybean cultivar Heinong 33 inoculated with Xinjiang soil were able to nodulate Heinong 33 and Williams showed very similar, or identical, plasmid, LPS and PCR-RAPD profiles. All the strains isolated from Xinjiang province, regardless of the soybean cultivar used for trapping, showed similar nodulation factor (LCO) profiles as judged by thin layer chromatographic analyses. These results indicate that the existence of soybean rhizobia sub-populations showing marked cultivar specificity, can affect the estimation of total soybean rhizobia populations indigenous to the soil, and can also affect the diversity of soybean rhizobial strains isolated from soybean nodules.

China↗

Determination of the chemical structure of the capsular polysaccharide of strain B33, a fast-growing soya bean-nodulating bacterium isolated from an arid region of China.

We have determined the structure of a polysaccharide from strain B33, a fast-growing bacterium that forms nitrogen-fixing nodules with Asiatic and American soya bean cultivars. On the basis of monosaccharide analysis, methylation analysis, one-dimensional 1H- and 13C-NMR and two-dimensional NMR experiments, the structure was shown to consist of a polymer having the repeating unit -->6)-4-O-methyl-alpha-D-Glcp-(1-->4)-3-O-methyl-beta-D-GlcpA-(1--> (where GlcpA is glucopyranuronic acid and Glcp is glucopyranose). Strain B33 produces a K-antigen polysaccharide repeating unit that does not have the structural motif sugar-Kdx [where Kdx is 3-deoxy-D-manno-2-octulosonic acid (Kdo) or a Kdo-related acid] proposed for different Sinorhizobium fredii strains, all of them being effective with Asiatic soya bean cultivars but unable to form nitrogen-fixing nodules with American soya bean cultivars. Instead, it resembles the K-antigen of S. fredii strain HH303 (rhamnose, galacturonic acid)n, which is also effective with both groups of soya bean cultivars. Only the capsular polysaccharide from strains B33 and HH303 have monosaccharide components that are also present in the surface polysaccharide of Bradyrhizobium elkanii strains, which consists of a 4-O-methyl-D-glucurono-L-rhamnan.

Carbohydrate Conformation↗

Structure-based mutagenesis reveals distinct functions for Ras switch 1 and switch 2 in Sos-catalyzed guanine nucleotide exchange.

Ras GTPases function as binary switches in signaling pathways controlling cell growth and differentiation. The guanine nucleotide exchange factor Sos mediates the activation of Ras in response to extracellular signals. We have previously solved the crystal structure of nucleotide-free Ras in complex with the catalytic domain of Sos (Boriack-Sjodin, P. A., Margarit, S. M., Bar-Sagi, D., and Kuriyan, J. (1998) Nature 394, 337-343). The structure demonstrates that Sos induces conformational changes in two loop regions of Ras known as switch 1 and switch 2. In this study, we have employed site-directed mutagenesis to investigate the functional significance of the conformational changes for the catalytic function of Sos. Switch 2 of Ras is held in a very tight embrace by Sos, with almost every external side chain coordinated by Sos. Mutagenesis of contact residues at the switch 2-Sos interface shows that only a small set of side chains affect binding, with the most important contact being mediated by tyrosine 64, which is buried in a hydrophobic pocket of Sos in the Ras.Sos complex. Substitutions of Ras and Sos side chains that are inserted into the Mg(2+)- and nucleotide phosphate-binding site of switch 2 (Ras Ala(59) and Sos Leu(938) and Glu(942)) have no effect on the catalytic function of Sos. These results indicate that the interaction of Sos with switch 2 is necessary for tight binding, but is not the critical driving force for GDP displacement. The structural distortion of switch 1 induced by Sos is mediated by a small number of specific contacts between highly conserved residues on both Ras and Sos. Mutations of a subset of these residues (Ras Tyr(32) and Tyr(40)) result in an increase in the intrinsic rate of nucleotide dissociation from Ras and impair the binding of Ras to Sos. Based on this analysis, we propose that the interactions of Sos with the switch 1 and switch 2 regions of Ras have distinct functional consequences: the interaction with switch 2 mediates the anchoring of Ras to Sos, whereas the interaction with switch 1 leads to disruption of the nucleotide-binding site and GDP dissociation.

Amino Acid Substitution↗

Microbial ecology of soils surrounding nuclear and thermal power plants in Taiwan.

This paper reports a study of the effect of three nuclear and one thermal power plants on the microbial ecology of soils. Populations of bacteria, actinomycetes, fungi, cellulolytic microbes, phosphate-solubilizing microbes and nitrogen-fixing microbes in the soil in the vicinity of each plant were studied. Soils were acidic at three sites, and moisture contents of the power plant soils were lower than those of the surrounding areas. Microbial populations of the topsoils (0-20 cm deep) were higher than the subsoils (21-40 cm deep), and only 10-15% of them showed significant difference (P < .05). Thirty-three percent of the samples from the surrounding areas had higher microbial population than those from the power plant areas, but 19% was the reverse. Populations of cellulolytic, phosphate-solubilizing and nitrogen-fixing microbes varied with sampling locations, season and environmental conditions. Ratios of cellulolytic, phosphate-solubilizing and nitrogen-fixing microbes to total viable counts in some samples of the surrounding areas were significantly higher than in the power plant areas. Although the microbial populations of power plant soil and its surrounding area were somewhat different, it cannot be attributed as an effect of power plant operation, as the differences were not consistent.

Environmental Monitoring↗

Natural product-based anti-HIV drug discovery and development facilitated by the NCI developmental therapeutics program.

During the decade 1987-1996, the Developmental Therapeutics Program (DTP) of the National Cancer Institute (NCI) provided infrastructure support for both intramural and extramural anti-HIV (human immunodeficiency virus) drug discovery research and development. This retrospective review describes some of the anti-HIV lead discovery and development that took place under DTP auspices or which was substantially facilitated by resources made available through the DTP. Examples highlighted include leads identified through the initial screening of pure natural product derived compounds and those derived from bioassay-guided fractionation of crude natural product extracts, and these are classified according to the mechanism of action targeting the critical steps within the replication cycle of HIV.

Amino Acid Sequence↗

The effect of jejunal meal feeding on gastroesophageal reflux.

BACKGROUND: Postprandial gastric distention is frequently associated with transient lower esophageal sphincter relaxation and gastroesophageal reflux (GER). Since the role of nutrient perfusion into the jejunum in inducing GER is not well understood, we studied the effect of jejunal feeding on GER through a percutaneous gastrojejunal tube in patients with and without reflux esophagitis. METHODS: Nine stroke patients with reflux esophagitis were fed through a percutaneous gastrojejunal tube with either a liquid meal (2 kcal/2 ml/min) or saline for 2 h randomly on 2 separate days. An esophageal pH probe was placed 5 cm above the gastroesophageal junction to detect acid reflux. Six stroke patients without esophagitis were enrolled as controls. RESULTS: In both the patients with esophagitis and the controls, esophageal acid exposure (15.3% (4.9%-28.2%) versus 2.7% (0.0%-10.8%), P=0.003; 5.9% (0.5%-6.7%) versus 0.0% (0.0%-1.5%), P = 0.01) and events of acid reflux (5 (1-16) versus 2 (0-8), P = 0.02; 12 (3-17) versus 1 (0-4), P = 0.02) were significantly greater during jejunal meal feeding than during saline infusion. Furthermore, in the reflux patients, but not in the controls, acid clearance time was also greater during jejunal meal feeding than during saline infusion (2.9 min (0.5-9.6 min) versus 0.7 min (0.0-4.3 min), P = 0.04). CONCLUSIONS: We therefore conclude that jejunal nutrient infusion without gastric distention can induce GER in both patients with reflux esophagitis and controls. This implies that GER induced by jejununal nutrients may in part explain the incapability of jejunal tube feeding to prevent gastropulmonary aspiration in patients at risk.

Aged↗

Small intestinal bacterial overgrowth versus antimicrobial capacity in patients with spontaneous bacterial peritonitis.

BACKGROUND: Spontaneous bacterial peritonitis (SBP) is a serious infection in cirrhotic patients with ascites. Both defects in the host defense mechanisms and the enhancement of the offensive factor (small intestinal bacterial overgrowth (SIBO)) may contribute to the development of SBP. Therefore, the aim of this study was to evaluate the role of SIBO versus various antimicrobial capacities in the pathogenesis of SBP in cirrhotic patients. METHODS: Forty-five cirrhotic patients were enrolled in this study. Bacterial overgrowth was evaluated by breath hydrogen test (BH2T). The hepatic reticuloendothelial system phagocytic index (HRESPI) was measured by intravenously injected colloid suspensions. RESULTS: The Child-Pugh scores in the SBP group were higher than in the non-SBP group (10.5 +/- 2.0 versus 8.0 +/- 1.8, P < 0.01). The ascitic protein concentration was significantly lower in the SBP group than in the non-SBP group (897 +/- 425 mg/l versus 1,325 +/- 453 mg/l, P < 0.01). Furthermore, the serum C3 concentration was lower in the SBP group than in the non-SBP group (43.1 +/- 13.6 ng/dl versus 73.2 +/- 26.4 ng/dl, P < 0.01). The serum C4 concentration was also lower in the SBP group than in the non-SBP group (12.4 +/- 4.0 ng/dl versus 16.9 +/- 6.6 ng/dl, P < 0.05). The incidence of SIBO was higher in the SBP group than in the non-SBP group (68.2% versus 17.4%, P < 0.01). HRESPI values were significantly higher in the two groups of cirrhotic patients than in the normal reference. However, there were no statistical differences in HRESPI between the two groups (8.4 +/- 2.8 min in the SBP group versus 7.9 +/- 2.8 min in the non-SBP group). CONCLUSIONS: The results of this study showed that the hepatic reticuloendothelial function is impaired in cirrhotic patients, but the degree of impairment does not differ between patients with and without previous history of SBP. Lower ascitic total protein, lower serum C3 and C4 concentrations, and presence of SIBO are all risk factors for SBP. Based on the results of our study, defects in the host defense mechanisms and the enhancement of the offensive factor (SIBO) may act in concert for the development of SBP.

Ascitic Fluid↗

Formation and regeneration of protoplasts for protease production in Streptomyces rimosus.

To improve the formation and regeneration frequency of protoplasts for protease production, experiments were performed using a cultivation of Streptomyces rimosus TM-55 (CCRC 940061) in a Tryptic-soy broth (TSB) containing 2% of glycine for 2 days. It was found that the protoplast formation decreased with increased incubation temperature and increased ratio of culture broth to vessel volume. The optimal incubation temperature was 28 degreesC and the ratio of culture broth to vessel volume was 2:5. The hypertonic medium containing 10 mM MgCl2, 25 mM CaCl2 and 500 mM sucrose provided high stability for protoplasts. Supplementation with MgSO4, KCl and NaNO3 improved the regeneration frequency of protoplasts. The smear method had a higher protoplast regeneration frequency than the pour plate method. Protoplasts had protease productivity which was similar to that obtained with fresh mycelia, with each milliliter of culture broth yielding 141 units of protease with 3.5 x 10(8) protoplasts and 148 units of protease with 14.25 mg fresh mycelia respectively in a shaking culture, while the values were 15 and eight units of protease in a static culture.

Agar↗

[Cloning of gene related to salt tolerance from Sinorhizobium fredii RT19 and its expression in Escherichia coli].

A 4.4 kb DNA fragment related to salt tolerance containing three open reading frames was isolated from the gene library of S. fredii strain RT19. By subcloning and functional analysis, only ORF2 related to salt tolerance was obtained. The ORF2 was ligated to expression vectors pThioHisA, B and C, respectively, and recombinant expression vectors pGA, pGB and pGC containing 1.5 kb DNA fragment related to salt tolerance were constructed. These recombinant expression vectors were transformed into E. coli DH5 alpha. Inducing by IPTG and analyzing with SDS-PAGE, it was found that the fusion protein encoded by pGC was expressed, and its molecular weight was equal to the sum of thioredoxin encoded by trxA and ORF2 putative protein molecular weight. The Western blot demonstrated that the target gene was successfully expressed in E. coli.

Cloning, Molecular↗

Reoperative Snodgrass procedure.

PURPOSE: We examine the results of a reoperative Snodgrass procedure for complex hypospadias. MATERIALS AND METHODS: A total of 25 patients with hypospadias in whom repair had failed underwent a reoperative Snodgrass procedure. Mean patient age and number of prior repairs were 11.4 years and 2.5, respectively. Before this treatment the meatus was at the distal shaft in 10 cases, mid shaft in 9, proximal shaft in 4 and penoscrotal junction in 2. Preoperatively the associated complications were fistulas in 13 patients, residual chordee in 12 and diverticulum in 1. In 8 cases the fistula was incorporated into the hypospadiac opening and treated as a longer defect. The urethral plates were, subjectively, surgically altered or unaltered in 7 and 18 patients, respectively. In 8 patients (group A) the previous repair(s) did not involve dissection of the urethral plate (unaltered) nor was there a fistula. The remaining 17 patients were classified as group B. The neourethra was then reconstructed as the Snodgrass technique. Followup urethral calibration was performed in 17 patients. RESULTS: Mean followup period was 13.7 months. Mean length and size of neourethra were 19.9 mm. and 14Fr, respectively. There were 7 (28%) postoperative fistulas of the neourethra. The incidences of postoperative fistula of the neourethra were 0% and 41.2% in groups A and B, respectively (p <0.01). Statistically the surgically altered urethral plate and the presence of a preoperative fistula were 2 independent factors predicting a higher postoperative fistula rate. The overall postoperative meatal stenosis rate was 13 (52%) with 3 (37.5%) in group A and 10 (58.8%) in group B, respectively (p >0.05). The meatal stenosis was treated with simple dilation in 11 cases and meatoplasty during subsequent fistula repair in 2. Stricture at the anastomostic site between the normal urethra and neourethra was noted in 2 patients. Fistula repair was successfully performed 6 months later in 4 patients. CONCLUSIONS: The Snodgrass procedure is a viable option for the treatment of previously failed hypospadias repair. It was highly effective in patients with a surgically unaltered urethral plate and no preoperative fistula.

Adolescent↗

Carbon dioxide flux density in cultivated rice paddy field.

The carbon dioxide flux density in a subtropical cultivated rice paddy field were determined over four crop seasons (two consecutive years) at five stages of rice plant development, transplanting, active tillering, booting, flowering and ripening, and an idle period between crops. During 24 h sampling periods at those stages, the CO2 flux densities were measured and found to be -2,000 to 55,000 mg CO2/m2/day in the first year and 600 to 29,000 mg CO2/m2/day in the second year in acrylic chambers covering rice plants. The CO2 flux densities in glass domes without plants but with soil were -700 to 6,000 mg CO2/m2/day in the first year and 147 to 4,500 mg CO2/m2/day in the second year. The data for the CO2 flux density over an entire year are extrapolated. Although our non-controlled environment experiments have inherent drawbacks, the collected data indicate that continuous rice cultivation has a tendency to result in increased CO2 emissions.

Carbon Dioxide↗

Endoscopic ultrasonography to study the causes of extragastric compression mimicking gastric submucosal tumor.

BACKGROUND AND PURPOSE: Many reports have confirmed that endoscopic ultrasonography (EUS) can differentiate gastric submucosal tumor from extragastric compression, but only a few specifically concentrated on EUS in identifying the causes of external compression. MATERIALS AND METHODS: From May 1993 to May 2001, we used EUS in 238 patients to diagnose gastric submucosal tumor or external compression. We excluded 183 patients who had submucosal tumors and analyzed the remaining 55 patients with extragastric compression. Malignant causes of external compression were proved by surgery or biopsy. Benign causes of external compression were proved by other imaging examinations (abdominal ultrasound, computerized tomography, angiography) or surgery. Patients with external compression caused by normal organs were followed up with repeated upper gastrointestinal endoscopy or EUS. RESULTS: The stomach was compressed by normal extragastric organs in 32 patients (spleen 10, splenic vessel 6, gall bladder 9, liver 3, pancreas 3, and intestine 1), by benign pathologic lesions in 12 patients (liver cyst 7, liver hemagioma 2, splenic cyst 1, pancreatic cyst 1, pancreatic cystadenoma 1) and by malignant tumors in 5 patients (hepatoma 1, liver metastasis from colon cancer 2, pancreatic cystadenocarcinoma 1 and lymphoma of spleen 1). In the remaining six patients, neither submucosal tumor nor external compression was found during EUS examination and the external compression was considered transient. CONCLUSION: When an extragastric compression mimicking submucosal tumor is detected by upper gastrointestinal endoscopy, EUS is indicated to identify the cause of extragastric compression.

Adolescent↗

Oxytetracycline production by immobilized Streptomyces rimosus.

This study examined whether the production of oxytetracycline by Streptomyces rimosus TM-55 (CCRC 960061) would be improved with calcium alginate immobilization in submerged fermentation compared with free cells. Results showed that in 1-mL culture broth, free cells produced 121 to 124 microg of oxytetracycline, whereas immobilized cells produced 153 to 252 microg. Immobilization of the cells retarded the growth rate of S. rimosus but increased the length of the growth period and improved the oxytetracycline production. The specific oxytetracycline productivity was 33.3 to 34.2 mg in each gram of free dry cells and was 40.2 to 40.7 mg in immobilized dry cells. The optimum immobilization conditions were alginate 2% and bead diameter of 2.13 mm. Oxytetracycline production increased with increasing inoculum density but decreased with increasing bead diameter. Ethylenediaminetetraacetic acid or monovalent ions could react with calcium in the bead or replace it with sodium ion, thereby reducing the strength of the beads.

Alginates↗

Expression of apoptosis-regulating proteins p53, Bcl-2, and Bax in primary resected esophageal squamous cell carcinoma.

Apoptosis plays a key role in the pathogenesis, aggressiveness, and therapy responsiveness of cancer. Proteins of the Bcl-2 family as well as p53 are important regulators of apoptosis. The present study retrospectively examines the expression of apoptosis-regulating proteins in primary resected esophageal squamous cell carcinoma (ESCC) and the correlation between the outcome of patients' treatment and the expression of the proteins. We used antibodies specific for the human p53, Bcl-2 and Bax proteins to examine the expression of these apoptosis-regulating proteins in 40 archival specimens of patients with primary resected esophageal squamous cell carcinoma. The overall expression of p53, Bcl-2, and Bax was 73%, 18%, and 100%, respectively. No significant correlations were found between the expression of p53, Bcl-2, and Bax. The expression of Bcl-2 had a negative influence on survival in this population of primary resected ESCC patients (p=0.03). But no differences in survival were observed in relation to the expression of p53 or Bax. In conclusion, Bcl-2 expression may provide additional and prognostic information for the clinical course of the disease and therefore to be developed as a prognostic indicator for primary resected esophageal squamous cell carcinoma.

Apoptosis↗

A second photochromic bacteriophytochrome from Synechocystis sp. PCC 6803: spectral analysis and down-regulation by light.

It now appears that photosynthetic prokaryotes and lower eukaryotes possess higher plant phytochrome-like proteins. In this work, a second phytochrome-like gene was isolated, in addition to the recently identified Cph1 phytochrome, from the Synechocystis sp. PCC 6803, and its gene product was characterized photochemically. The open reading frame sll0821 (designated cph2 in this work) has structural characteristics similar to those of the plant phytochromes and the Synechocystis Cph1 with high amino acid sequence homology in the N-terminal chromophore binding domain. The predicted Cph2 protein consists of 1276 amino acids with a calculated molecular mass of 145 kDa. Interestingly, the Cph2 protein has two putative chromophore binding domains, one around Cys-129 and the other around Cys-1022. The Cph2 was overexpressed in E. coli as an Intein/CBD (chitin binding domain) fusion and in vitro reconstituted with phycocyanobilin (PCB) or phytochromobilin (PPhiB). Both the Cph2-PCB and Cph2-PPhiB adducts showed the typical photochromic reversibility with the difference spectral maxima at 643/690 and 655/701 nm, respectively. The Cys-129 was confirmed to be the chromophore binding residue by in vitro mutagenesis and Zn(2+) fluorescence. The microenvironment of the chromophore in Cph2 seems to be similar to that in plant phytochromes. The cph2 gene expression was dark-induced and down-regulated to a basal level by light, like the cph1 gene. These observations suggest that Synechocystis species have multiple photosensory proteins, probably with distinct roles, as in higher plants.

Amino Acid Sequence↗