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Biomedical subjects

C S Chen

Publications and source records attributed to C S Chen.

At least 55 records · Page 3Linked to original sources
PubMed2000

Overexpression of the soluble form of chicken cystatin in Escherichia coli and its purification.

A cDNA encoding chicken cystatin was cloned into the pET-23a(+) expression vector and then transformed into Escherichia coli AD494(DE3)pLysS expression host. An active soluble form of cystatin was expressed in the cytoplasm of E. coli induced by isopropyl beta-D-thiogalactopyranoside. The recombinant chicken cystatin was purified to electrophoretic homogeneity by a simple and rapid method involving heat treatment and Sephacryl S-100 gel filtration chromatography. The recombinant cystatin behaved as a thermal-stable protein and exhibited papain-like protease inhibition activity comparable to the natural chicken cystatin.

Animals↗
PubMed2000

E-cadherin expression in surgically-resected non-small cell lung cancers--a clinicopathological study.

BACKGROUND: E-Cadherin is a subclass of the cadherin family that plays a major role in the maintenance of intercellular junctions in normal epithelium. Decreased expression of E-cadherin might be closely related to invasiveness and dedifferentiation in human cancers. This study is aimed at investigating the clinicopathological significance of E-cadherin expression and its impact on the prognosis in surgically resected non-small-cell lung cancer patients. METHODS: Using immunohistochemical staining, the expression of E-cadherin was studied in 207 surgically resected lung cancer specimens from January 1990 through December 1994. The clinicopathological data and survival status were recorded and analysed against the E-cadherin expression level in each tumor. RESULTS: E-cadherin expression was detected in 122 of the 207 lung tumors (59.0%), and the expression was significantly lower in tumors with poor differentiation (p < 0.001), in tumors with vascular invasion (p < 0.05), and in tumors with direct invasion into surrounding structures (p < 0.01). There was no correlation between E-cadherin expression, and tumor stage and regional lymph-node metastasis. There was no significant difference in survival rate between higher (> 40%) and lower (< 40%) E-cadherin expression groups; however, in tumors 3 cm or less, a significant difference was found between higher and lower E-cadherin expressions (p < 0.0001). CONCLUSIONS: Underexpression of E-cadherin is associated with poor differentiation and invasiveness in NSCLC. In patients with small NSCLC (< or = 3 cm), higher E-cadherin expression (> 40%) significantly had a favorable prognosis.

Adult↗
PubMed2000

Successful treatment of recurrent pelvic desmoid tumour with tamoxifen: case report.

The case report of a young woman with recurrent pelvic desmoid tumour successfully treated with tamoxifen is described. The desmoid tumour recurred within 6 months after the initial exploratory laparotomy. Tamoxifen therapy led to complete relief of ascites within 2 months and complete tumour regression by the end of the fourth month, and the patient has remained stable for 6 years. Without sacrificing pelvic organs or major vessels and preserving reproductive ability, tamoxifen should be considered as the first drug of choice in such a recurrent condition.

Adult↗
PubMed2000

Immunolocalization of macrophages and transforming growth factor-beta 1 in induced rat periapical lesions.

Apical periodontitis was induced in Wistar rats by exposing the pulp chamber of right mandibular first molars to the oral environment. Animals were killed 0, 5, 10, 15, 20, 30, 60, and 80 days after lesion induction. Microradiographic and automated image analysis showed that the lesions expanded significantly in a time-dependent manner from day 0 to day 20 (0.039 mm2/day, p < 0.05, active phase) and stabilized thereafter (chronic phase). A linear regression test revealed a positive correlation between the numbers of ED-1 positive macrophage per microscopic high power field and the periapical lesion size during the active phase (r = 0.98, p < 0.01). Immunohistochemical studies showed that transforming growth factor-beta 1 positive macrophages distributed around the root apex and areas showing bone resorption during active lesion phase, whereas TGF-beta 1-positive osteoblasts were detected during the chronic stage (days 30, 60, and 80 after pulp exposure). Histologically TGF-beta 1 positive osteoblasts possessed a large, round nucleus as well as an abundant cytoplasm and located in close vicinity to areas exhibiting reparative bone formation. These results suggest that macrophages may play important role(s) in the initiation and development of periapical lesions and TGF-beta 1 may play dual roles in both bone resorption and deposition in induced rat periapical lesions.

Acute Disease↗
PubMed2000

Characterization of the distal tail fiber locus and determination of the receptor for phage AR1, which specifically infects Escherichia coli O157:H7.

Phage AR1 is similar to phage T4 in several essential genes but differs in host range. AR1 infects various isolates of Escherichia coli O157:H7 but does not infect K-12 strains that are commonly infected by T4. We report here the determinants that confer this infection specificity. In T-even phages, gp37 and gp38 are components of the tail fiber that are critical for phage-host interaction. The counterparts in AR1 may be similarly important and, therefore, were characterized. The AR1 gp37 has a sequence that differs totally from those of T2 and T4, except for a short stretch at the N terminus. The gp38 sequence, however, has some conservation between AR1 and T2 but not between AR1 and T4. The sequences that are most closely related to the AR1 gp37 and gp38 are those of phage Ac3 in the T2 family. To identify the AR1-specific receptor, E. coli O157:H7 was mutated by Tn10 insertion and selected for an AR1-resistant phenotype. A mutant so obtained has an insertion occurring at ompC that encodes an outer membrane porin. To confirm the role of OmpC in the AR1 infection, homologous replacement was used to create an ompC disruption mutant (RM). When RM was complemented with OmpC originated from an O157:H7 strain, but not from K-12, its AR1 susceptibility was fully restored. Our results suggest that the host specificity of AR1 is mediated at least in part through the OmpC molecule.

Bacteriophages↗
PubMed2000

Modified technique for introducer-type percutaneous endoscopic gastrostomy and jejunostomy: assisted by a novel trocar and fasteners.

BACKGROUND AND PURPOSE: Commercially available kits for percutaneous endoscopic gastrostomy (PEG) and jejunostomy comprise a substantial proportion of the cost of patients for this procedure. A modified introducer-type technique and new instrumentation for PEG and jejunostomy that substantially reduces the cost of the kit were tested for efficacy and safety. METHODS: This technique was tested on 10 pigs as a pilot study, and then applied to seven consecutive patients undergoing gastrostomy and three patients undergoing gastrostomy with jejunostomy. The endoscopy and site selection for gastrostomy were the same as in standard PEG. Two novel fasteners for fixing the gastric wall to the abdominal wall and a guide-wire in the selected site were inserted separately into the stomach through 15-gauge needles. A stainless steel trocar with a detachable sheath was introduced into the stomach over the guide-wire. After the inner stylet was removed, a 24-French Foley catheter was inserted as a feeding tube. Gastrostomy was completed after balloon inflation and external fixation. If jejunostomy was indicated, a 12-French nasogastric tube was inserted through the gastrostomy. The procedure time, complications, and costs were compared with those for another 15 consecutive patients who underwent the conventional pull-through method of PEG and jejunostomy using commercially available kits. RESULTS: No significant difference was found in procedure time between patients who underwent the modified or conventional gastrostomy procedures (mean +/- standard deviation, 15.4 +/- 5.6 min). There was a similar incidence of short-term complications in the two treatment groups. The feeding catheters required replacement more quickly than did those in the commercial kits (80 +/- 58 vs 217 +/- 140 d). The cost to patients was much less with the new method than with conventional PEG. CONCLUSIONS: The new gastrostomy method achieves the same medical quality at far less cost for patients.

Adult↗
PubMed2000

Validation of the Chinese Health Questionnaire (CHQ-12) in community elders.

There has been no validated screening instrument for use in non-psychotic illness of the elderly in Taiwan. This study aims to test the validity of the 12-item Chinese Health Questionnaire (CHQ-12) among the elderly in a community study. The CHQ-12 was administrated via reading-out to 222 subjects aged 65 and over from three communities. Psychiatrists using the Geriatric Mental Status Schedule (GMS) assessed psychiatric condition while the diagnosis was made according to the computerized program, AGECAT. Validity indices of the CHQ-12 were calculated, using the Relative Operating Characteristic (ROC) analysis for its optimal cut-off point. Variables hypothesized to affect its performance were assessed. Validity of the CHQ-12 at optimal cut-off point 2/3 were estimated with a sensitivity of 79.7%, a specificity of 83.6%, a positive predictive value of 68.9%, a negative predictive value of 90.0% and an overall misclassification rate of 17.6%, and an estimate of the area under the ROC curve of 0.81. The performance of the CHQ-12 was better in males, in those who were literate, and in those without any physical illness. This study demonstrated that the use of CHQ-12 in the elderly community is as valid as in the general population survey. However, it should be read out by the investigator rather than self-administered due to the high proportion of illiteracy among the Taiwanese elderly.

Aged↗
PubMed2000

Coronary artery bypass grafting on the beating heart using the octopus method--a case report.

A 74-year-old male was admitted due to chest tightness for one month. He had received percutaneous transilluminal coronary angioplasty (PTCA) because of single-vessel disease one year ago. Cardiac catheterization examination carried out during this admission revealed 90% stenosis of the left anterior descending coronary artery (LAD) and circumflex branch (CX). Because another attempt of PTCA was considered not optimal, the patient was advised to undergo surgical grafting to which he consented. After expediently balancing the merits and demerits of every practicable surgical procedure--the essential determinant in selection of which was that the patient's condition and criteria of indication of that procedure were in perfect harmony--we decided to carry out minimally invasive direct coronary artery bypass (MIDCAB) with the application of Octopus tissue stabilizer. We report the surgical course and anesthetic management of the patient and discourse some detail in MIDCAB.

Aged↗
PubMed2000

The role of free to total prostate-specific antigen ratio for prostate cancer in screening patients with total serum levels between 4 and 20 ng/ml.

BACKGROUND: We retrospectively tried to determine if the free to total prostate-specific antigen (f/t PSA) ratio could improve the specificity of PSA in prostate cancer screening of patients with total serum levels between 4 and 20 ng/ml. METHODS: Two hundred ninety-five patients with serum PSA levels from 4 to 20 ng/ml had undergone sextant prostate needle biopsy. Each patient had no prior history of prostate cancer, acute urine retention, or prostatitis. Prebiopsy free PSA values were measured in 155 patients. Total PSA levels were determined with the AxSYM enzyme-linked immunosorbent assay. Free PSA levels were measured with the AxSYM microparticle enzyme immunoassay. RESULTS: Mean f/t PSA ratios were 0.114+/-0.004 in men of the cancer group and 0.161+/-0.008 in men of the benign group (p<0.002). Based on the analysis of sensitivity and specificity in relation to f/t PSA ratios, use of the 18% cutoff point could detect 89% of cancer cases, and at the same time could avoid 35% of unnecessary prostate biopsies. The areas under the receiver-of-characteristic curve for f/t PSA ratio and total PSA were 0.649 and 0.545, respectively. CONCLUSION: Serum f/t PSA ratios were significantly lower in patients with prostate cancer than in patients with benign disease. The determination of an appropriate f/t PSA ratio should be based on the generated data such as that demonstrated in this study in order to improve diagnostic accuracy and specificity for patients with equivocal PSA values and to avoid conducting unnecessary prostate biopsies.

Aged↗
PubMed1999

Enzymatic catalysis in cosolvent modified pressurized organic solvents.

An important advantage of carrying out enzymatic catalysis in organic media is the increased solubility of hydrophobic substrates. This study compares a model lipase catalyzed esterification of cholesterol using vinyl acetate (VA) in two such nontraditional media: high-pressure hexane and supercritical (SCF) ethane. The effect of using one of the reactants (VA) as a cosolvent to increase the solubility of the other reactant (cholesterol) in SCF ethane has been investigated. The thermodynamic activity of water (a(w)) in the reaction media was controlled by the direct addition of the salt hydrate pair Na(4)P(2)O(7)/Na(4)P(2)O(7).10H(2)O. The a(w) of the salt hydrate system is shown to be a function of pressure and its variation over the pressure range 104-173 bar has been estimated. The initial reaction rate in pressurized hexane was found to vary linearly with the cholesterol concentration. The reaction rate was also a function of pressure-the effect being more pronounced in ethane than in hexane. This is consistent with the large negative partial molar volumes observed in SCFs, although the sign of the resulting activation volume differs from previous investigations of lipase-catalyzed reactions in SCFs. When corrected for substrate concentration, the initial rate of catalysis in SCF ethane was determined to be greater than in pressurized hexane over the conditions investigated. This study shows that proper solvent choice can be used to regulate reaction rates in pressurized solvents.

Catalysis↗
PubMed1999

Broad screening test for sphingolipid-storage diseases.

BACKGROUND: Lipid-storage diseases are collectively important because they cause substantial morbidity and mortality, and because they may present as dementia, major psychiatric illness, developmental delay, or cerebral palsy. At present, no single assay can be used as an initial general screen for lipid-storage diseases. METHODS: We used a fluorescent analogue of lactosylceramide, called N-[5-(5,7-dimethylborondipyrromethenedifluoride)-1-pentanoyl]D- lactosylsphingosine (BODIPY-LacCer), the emission of which changes from green to red wavelengths with increasing concentrations in membranes, to examine the intracellular distribution of the lipid within living cells. FINDINGS: During a brief pulse-chase experiment, the fluorescent lipid accumulated in the lysosomes of fibroblasts from patients with Fabry's disease, GM1 gangliosidosis, GM2 gangliosidosis (Tay-Sachs and Sandhoff forms), metachromatic leucodystrophy, mucolipidosis type IV, Niemann-Pick disease (types A, B, and C), and sphingolipid-activator-protein-precursor (prosaposin) deficiency. In control cells, the lipid was mainly confined to the Golgi complex. In a masked study, replicate samples of 25 of 26 unique cell lines representing ten different lipid-storage diseases, and 18 of 20 unique cell lines representing controls were correctly identified; the sensitivity was 96.2% (95% CI 80.4-99.9) and the specificity 90.0% (68.3-98.8). INTERPRETATION: This method may be useful as an initial general screen for lipid-storage diseases, and, with modification, could be used for large-scale automated screening of drugs to abrogate lysosomal storage in various lipidoses. The unexpected accumulation of BODIPY-LacCer in several biochemically distinct diseases raises important questions about common mechanisms of cellular dysfunction in these disorders.

Antigens, CD↗
PubMed1999

Identification of multiple phosphoinositide-specific phospholipases D as new regulatory enzymes for phosphatidylinositol 3,4, 5-trisphosphate.

In the course of delineating the regulatory mechanism underlying phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) metabolism, we have discovered three distinct phosphoinositide-specific phospholipase D (PI-PLD) isozymes from rat brain, tentatively designated as PI-PLDa, PI-PLDb, and PI-PLDc. These enzymes convert [3H]PI(3,4,5)P3 to generate a novel inositol phosphate, D-myo-[3H]inositol 3,4,5-trisphosphate ([3H]Ins(3,4,5)P3) and phosphatidic acid. These isozymes are predominantly associated with the cytosol, a notable difference from phosphatidylcholine PLDs. They are partially purified by a three-step procedure consisting of DEAE, heparin, and Sephacryl S-200 chromatography. PI-PLDa and PI-PLDb display a high degree of substrate specificity for PI(3,4, 5)P3, with a relative potency of PI(3,4,5)P3 >> phosphatidylinositol 3-phosphate (PI(3)P) or phosphatidylinositol 4-phosphate (PI(4)P) > phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) > phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2). In contrast, PI-PLDc preferentially utilizes PI(3)P as substrate, followed by, in sequence, PI(3,4,5)P3, PI(4)P, PI(3,4)P2, and PI(4,5)P2. Both PI(3, 4)P2 and PI(4,5)P2 are poor substrates for all three isozymes, indicating that the regulatory mechanisms underlying these phosphoinositides are different from that of PI(3,4,5)P3. None of these enzymes reacts with phosphatidylcholine, phosphatidylserine, or phosphatidylethanolamine. All three PI-PLDs are Ca2+-dependent. Among them, PI-PLDb and PI-PLDc show maximum activities within a sub-microM range (0.3 and 0.9 microM Ca2+, respectively), whereas PI-PLDa exhibits an optimal [Ca2+] at 20 microM. In contrast to PC-PLD, Mg2+ has no significant effect on the enzyme activity. All three enzymes require sodium deoxycholate for optimal activities; other detergents examined including Triton X-100 and Nonidet P-40 are, however, inhibitory. In addition, PI(4,5)P2 stimulates these isozymes in a dose-dependent manner. Enhancement in the enzyme activity is noted only when the molar ratio of PI(4,5)P2 to PI(3,4, 5)P3 is between 1:1 and 2:1.

Animals↗
PubMed1999

Geometric control of switching between growth, apoptosis, and differentiation during angiogenesis using micropatterned substrates.

Past studies using micropatterned substrates coated with adhesive islands of extracellular matrix revealed that capillary endothelial cells can be geometrically switched between growth and apoptosis. Endothelial cells cultured on single islands larger than 1500 microm2 spread and progressed through the cell cycle, whereas cells restricted to areas less than 500 microm2 failed to extend and underwent apoptosis. The present study addressed whether island geometries that constrained cell spreading to intermediate degrees, neither supporting cell growth nor inducing apoptosis, cause cells to differentiate. Endothelial cells cultured on substrates micropatterned with 10-microm-wide lines of fibronectin formed extensive cell-cell contacts and spread to approximately 1000 microm2. Within 72 h, cells shut off both growth and apoptosis programs and underwent differentiation, resulting in the formation of capillary tube-like structures containing a central lumen. Accumulation of extracellular matrix tendrils containing fibronectin and laminin beneath cells and reorganization of platelet endothelial cell adhesion molecule-positive cell-cell junctions along the lengths of the tubes preceded the formation of these structures. Cells cultured on wider (30-microm) lines also formed cell-cell contacts and aligned their actin cytoskeleton, but these cells spread to larger areas (2200 microm2), proliferated, and did not form tubes. Use of micropatterned substrates revealed that altering the geometry of cell spreading can switch endothelial cells among the three major genetic programs that govern angiogenesis-growth, apoptosis and differentiation. The system presented here provides a well-defined adhesive environment in which to further investigate the steps involved in angiogenesis.

Animals↗
PubMed1999

Use of N-[5-(5,7-dimethyl boron dipyrromethene difluoride-sphingomyelin to study membrane traffic along the endocytic pathway.

We have used N-[5-(5,7-dimethyl boron dipyrromethene difluoride)-1-pentanoyl]-D-erythro-sphingosylphosphorylcholine (C5-DMB-SM or 'BODIPY-SM'), a fluorescent analog of sphingomyelin, to study lipid transport along the endocytic pathway of human skin fibroblasts. The unique spectral properties of the BODIPY fluorophore allow the investigator to distinguish various populations of labeled endosomes and lysosomes within the living cell by fluorescence microscopy, and in conjunction with quantitative fluorescence microscopy, to estimate the concentration of these lipids in different intracellular compartments. This methodology is also applicable for visualizing the accumulation of lipids in the endosomes and lysosomes of storage disease fibroblasts.

Biological Transport↗
PubMed1999

Elevated lysosomal pH in Mucolipidosis type IV cells.

The lysosomal pH in Mucolipidosis type IV (ML-IV) and several other storage disease fibroblasts (Niemann Pick, type A; Niemann Pick, type C; Hunter (MPS II); and Farber) and in normal human skin fibroblasts was determined in situ. Cells were pulse labeled with a fluorescein-conjugated dextran to label the lysosomes. Quantitative fluorescence microscopy was then carried out on living cells to measure the ratio of fluorescence at two different excitation wavelengths. An image processing routine was used to quantify fluorescence from individual lysosomes. Ratiometric data were converted to an absolute value of pH using an appropriate standard curve. Lysosomal pH varied between 4.3 and 4.5 for all the cell types examined except ML-IV cells which was almost one pH unit higher (pH approximately 5.2). Qualitatively similar results were obtained using acridine orange, another fluorophore whose fluorescence emission is pH dependent, ruling out the possibility that the stored molecules in ML-IV cells might induce an artifact in the fluorescein-based pH measurements. We conclude that elevated lysosomal pH is unique to ML-IV cells. This property may be an important factor, if not the cause, for the accumulation of the broad spectrum of substances, including sphingolipids, phospholipids, and acid mucopolysaccharides, even though the lysosomal hydrolases participating in the catabolism of these molecules appear to be normal.

Humans↗