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Biomedical subjects

L S Chang

Publications and source records attributed to L S Chang.

At least 91 records · Page 5Linked to original sources
PubMed1996

Chemical modification of cationic residues in toxin a from king cobra (Ophiophagus hannah) venom.

The cationic groups of arginine and lysine residues in alpha-neurotoxin, Toxin a, isolated from king cobra (Ophiophagus hannah) venom were subjected to modification with trinitrobenzene sulfonate (TNBS) and p-hydroxyphenylglyoxal (HPG), respectively. The trinitrophenylated (TNP) derivatives of Toxin a at Lys-10, 56, or 71 showed approximately 25% residual lethality, and modifications on Lys-10 and 56 or Lys-10 and 50 resulted in a decrease of lethality by 84% and 86%, respectively. Modifications on Arg-34, 37, and 70 and Arg-34, 37, and 72 in Toxin a caused a decrease in lethality by 92% and 93%, respectively, and it almost completely lost its lethality and binding activity to nicotinic acetylcholine receptor (nAChR) when all four arginine residues were modified. These results indicate that in addition to the cationic residues on loop II (Arg-34, 37), loop III (Lys-50, 56), and the C-terminal tail (Arg-70, 72; Lys-71), Lys-10 on loop I is also related to the neurotoxicity of Toxin a.

Amino Acid Sequence↗
PubMed1996

Chemical modification of notexin from Notechis scutatus scutatus (Australian tiger snake) venom with pyridoxal-5'-phosphate.

Notexin from Notechis scutatus scutatus snake venom was subjected to Lys modification with pyridoxal 5'-phosphate (PLP), and one major modified derivative was purified on a cation-exchanger SP-8HR column. The results of amino acid analysis and sequence determination revealed that only 2 Lys residues at positions 82 and 115 out of 11 Lys residues in notexin were modified. The incorporation of PLP into the protein was accompanied by the loss of 53% lethal toxicity, but the modified notexin showed an about 1.2-fold increase in enzymatic activity. However, the secondary structure of the toxin molecule did not significantly change after modification with PLP as revealed by the CD spectra, and the antigenicity of PLP derivative remained unchanged. The modified derivative retained its affinity for Ca2+, indicating that the modified Lys residues did not participate in Ca2+ binding. These results indicate that modification of Lys residues causes a differential effect on the enzymatic activity and lethal toxicity of notexin, and suggest that notexin might possess two functional sites, one responsible for the catalytic activity and the other associated with its lethal effect.

Amino Acid Sequence↗
PubMed1996

The essentiality of His-47 and the N-terminal region for the binding of 8-anilinonaphthalene-1-sulfonate with Taiwan cobra phospholipase A2.

The binding of the apolar fluorescent dye 8-anilinonaphthalene-1-sulfonate (ANS) to Naja naja atra phospholipase A2 (PLA2) as well as the enhancement of ANS fluorescence of the PLA2-ANS complex decreased with increasing pH in a pH range from 3 to 9. These pH-dependent curves can be well interpreted as the perturbation of an ionizable group with pK value of 5.8, which was assigned as His-47 in the active site of PLA2. The ionizable group with pK 5.8 was no longer observed after methylation of His-47, supporting the idea that the pH dependence of ANS binding arose from an electrostatic interaction between His-47 and the bound ANS. Removal of the N-terminal octapeptide of PLA2 caused a precipitous drop in the capability of PLA2 for binding with ANS and enhancing ANS fluorescence, reflecting that the integrity of the N-terminal region was essential for maintaining the hydrophobic character of the ANS-binding site. However, the nonpolarity of the ANS-binding site in the N-terminus-removed derivative was still partially retained at low pH, but was completely lost at high pH. Evidently, the N-terminal region plays a more crucial role in ANS binding at high pH than at low pH. These results indicate that hydrophobic interaction as well as electrostatic interaction are involved in the binding of ANS to PLA2, and that the relative contributions of both interactions in ANS fluorescence enhancement may be different under different pH.

Anilino Naphthalenesulfonates↗
PubMed1996

The functional involvement of Lys-38 in the heavy subunit of rat kidney gamma-glutamylcysteine synthetase: chemical modification and mutagenesis studies.

Rat kidney gamma-glutamylcysteine synthetase (gamma GCS) was inactivated by reaction with trinitrobenzene sulfonate (TNBS), and the reaction followed pseudo-first-order kinetics. Inactivation kinetics revealed that only one of the amino acid residues modified by TNBS was essential for gamma GCS activity. The addition of 10 mM Mg2+ to the TNBS inactivation reaction resulted in a 16-fold increase in the rate of inactivation. Chromatographic analysis on the tryptic hydrolyzates of trinitrophenylated (TNP) derivatives showed that Lys-38 in the gamma GCS heavy subunit was significantly modified in the presence of Mg2+. In contrast to small changes in the catalytic properties observed by mutation of Lys-38 to Arg, the mutants K38N and K38E had a marked decrease in enzymatic activity and about twofold increase in Km for glutamate. These results suggest that the positively charged Lys-38 may be involved in the binding of glutamate to gamma GCS.

Adenosine Triphosphate↗
PubMed1996

The essentiality of calcium ion in the enzymatic activity of Taiwan cobra phospholipase A2.

In order to address the mechanism whereby Ca2+ wad crucial for the manifestation of the enzymatic activity of phospholipase A2 (PLA2), four divalent cations were used to assess their influences on the catalytic activity and the fine structures of Naja naja atra PLA2. It was found that substitution of Mg2+ or Sr2+ for Ca2+ in the substrate solution caused a decrease in the PLA2 activity to 77.5% or 54.5%, respectively, of that in the presence of Ca2+. However, no PLA2 activity was observed with the addition of Ba2+. With the exception of Mg2+, the nonpolarity of the 8-anilinonaphthalene-1-sulfonate (ANS)-binding site of PLA2 markedly increased with the binding of cations to PLA2. In the meantime, the accessibilities of Lys-6 (65) and Tyr-3 (63) toward trinitrobenzene sulfonate and p-nitrobenzenesulfonyl fluoride were enhanced by the addition of Ca2+, Sr2+, and Ba2+, but not by Mg2+. The order of the ability of cations to enhance the ANS fluorescence and the reactivity of Lys and Tyr residues toward modified reagents was Ba2+ > Sr2+ > Ca2+ > Mg2+, which was the same order as the increase in their atomic radii. These results, together with the observations that the ANS molecule binds at the active site of PLA2 and that Tyr-3, Lys-6, and Tyr-63 of PLA2 are involved in the binding with the substrate, suggest that the binding of Ca2+ to PLA2 induces conformational changes at the active site and substrate-binding site. However, the smaller atomic radius with Mg2+ or the bigger atomic radii with Sr2+ and Ba2+ might render the conformation improperly rearranged after their binding to PLA2 molecule.

Anilino Naphthalenesulfonates↗
PubMed1996

cDNA sequence analysis and mutagenesis studies on the A chain of beta-bungarotoxin from Taiwan banded krait.

The cDNA encoding the A chain of beta-bungarotoxin (beta-Bgt) was constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus (Taiwan banded krait). The deduced amino acid sequence encoding the A chain revealed that the determined chain was different from the known A chains (A1, A2, A3, A4, and A5). Nevertheless, the amino acid sequence and the cDNA sequence of the novel A chain were highly homologous with those of other A chains. The gene encoding the A chain of beta-Bgt was subjected to mutagenesis, and the Tyr-11, Cys-15, and Leu-72 of the A chain were substituted by Cys-11, Ser-15, and Cys-72, respectively. Instead of the six disulfide bonds observed with the A chain, the resulting mutant contained seven disulfide linkages in its molecular structure which simulated those of presynaptic PLA2 neurotoxins and PLA2 enzymes. However, the mutant did not exhibit a higher phospholipase activity than that noted with the recombinant A chain. These results seem to suggest that, in the absence of the B chain, the six pairs of disulfide bonds in the recombinant A-chain molecule are enough to maintain its active conformation for exerting the phospholipase activity.

Amino Acid Sequence↗
PubMed1996

Alterations in pulmonary function after retroperitoneoscopic surgery.

OBJECTIVE: To evaluate and compare changes in pulmonary function after retroperitoneoscopic and open surgery. PATIENTS AND METHODS: From June 1994 to October 1995. 11 patients (five men and six women, mean age 44.7 years, range 29-69) underwent retroperitoneoscopic procedures (Group 1) and 11 patients (eight men and three women, mean age 57.5 years, range 22-73) underwent flank-incision procedures (Group 2). The surgery comprised eight adrenalectomies and one each of nephroureterectomy, nephrectomy and pyelolithotomy in Group 1, and five adrenalectomies, four nephroureterectomies, one ureterolithotomy and one excision of a retroperitoneal tumour in Group 2. Pulmonary function tests (PFTs) were performed before and 3 days after surgery; the forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), forced expiratory volume at 25% (FEV25%), FEV1/FVC, vital capacity (VC), total lung capacity (TLC), residual lung volume (RV) and functional residual capacity (FRC) were compared between the groups. The post-operative changes in the PFTs were assessed using a paired t-test and the degree of change in both groups compared using the Mann-Whitney U-test. Other factors possibly influencing PFT were analysed using multiple regression. RESULTS: Pulmonary function was impaired in both groups on the third day after surgery. In Group 2, the FVC, FEV1, FEV25%, VC and TLC had declined significantly (all P < 0.05) from the pre-operative value. In Group 1, only the FVC and VC decreased significantly (P < 0.05). Post-operative pulmonary complications occurred in two patients in Group 2 but in none of those in Group 1, showing that pulmonary function was generally less affected in Group 1 than in Group 2. CONCLUSION: Although pulmonary function was impaired 3 days after surgery in both groups, retroperitoneoscopic surgery, by eliminating a large flank incision, caused less post-operative pulmonary dysfunction than open surgery.

Adult↗
PubMed1996

The significance of tumour grade in predicting disease progression in stage Ta transitional cell carcinoma of the urinary bladder.

OBJECTIVE: To determine the significance of tumour grade in predicting disease progression in stage Ta transitional cell carcinoma (TCC) of the urinary bladder. PATIENTS AND METHODS: From August 1975 to January 1991, 140 patients (121 men and 19 women, mean age 64.2 years, range 30-83) with stage Ta TCC of the urinary bladder were treated at the Veterans General Hospital-Taipei by transurethral resection and post-operative adjuvant intravesical chemotherapy. There were 48 patients (34%) with grade 1, 78 patients (56%) with grade 2 and 14 patients (10%) with grade 3 tumours. Disease progression and muscle invasion were assessed during a mean follow-up of 74.3 months and related to grade, urine cytology and the number and location of tumours. RESULTS: A total of 10 patients (7%) had tumour recurrence which invaded the muscle, of whom one had grade 1, six had grade 2 and three had grade 3 tumours. The patient with a grade 1 tumour developed muscle invasion within 16 months of surgery. The mean interval to the development of muscle invasion was 49.2 months in patients with grade 2 and 39.4 months with grade 3 tumour. In patients with stage Ta TCC, a grade 3 tumour was more likely to progress to muscle invasion (21%) than was a grade 1 (2.1%; P < 0.05) or grade 2 (7.7%; P < 0.05) tumour. One patient with a grade 2 and one patient with grade 3 tumour developed distant metastases without muscle invasion. Overall, the disease progression rate was 8.6% (12/140) in stage Ta TCC. Disease progression was more common with grade 3 (28.5%) than with grade 2 (9.0%; P < 0.05) and grade 1 (2.1%; P < 0.05) tumours. CONCLUSION: Tumour grade may be used to predict disease progression after resection of stage Ta urinary bladder cancer. A higher grade of tumour was associated with a higher incidence of disease progression.

Administration, Intravesical↗
PubMed1996

The interaction of 8-anilinonaphthalene-1-sulfonate with His-47 of Taiwan cobra phospholipase A2 perturbing by the binding of calcium ion.

The 8-anilinonaphthalene-1-sulfonate (ANS) fluorescence intensity of ANS-Naja naja atra (Taiwan cobra) phospholipase A2 (PLA2) complex increased with the addition of Ca2+, but the observed fluorescence enhancement markedly decreased after methylation of His-47 in PLA2 molecule. However, the binding affinities of methylated PLA2 for ANS and Ca2+ were similar to or even greater than those observed with native PLA2. These results, together with the finding that ANS electrostatically interacted with His-47 of PLA2, suggest that the increase in the intensity of ANS fluorescence upon the addition of Ca2+, in part, arises from the ionic interaction of His-47 with ANS being perturbed by the binding of Ca2+.

Anilino Naphthalenesulfonates↗
PubMed1996

Structural determinants of cobrotoxin affect its mobility on sodium dodecyl sulfate polyacrylamide gel.

In order to identify the structural determinants of cobrotoxin that caused its anomalous mobility in sodium dodecyl sulfate (SDS) gel, comparative studies on the capacities of cobrotoxin and its modified derivatives for binding with SDS were made. Cobrotoxin migrates faster in SDS gel after modification of Lys residues or carboxylated groups. However, the capacities of modified derivatives for binding with SDS molecules did not show a positive correlation with their electrophoretic mobilities. Moreover, it was found that the segment at positions 22-38 had the greatest capacity to bind with SDS than other segments of cobrotoxin, but guanidination of Lys residues of this peptide fragment did not significantly change its migration. These results suggest that the SDS-binding capacity of cobrotoxin is not solely responsible for its anomalous mobility in SDS gel, and that the effects may arise from the electrostatic interactions between Lys residue and carboxylated groups.

Cobra Neurotoxin Proteins↗
PubMed1996

cDNA sequence analysis of a novel cardiotoxin-like protein from Taiwan banded krait.

The cDNAs encoding a novel protein was constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus by reverse transcription-polymerase chain reaction. The deduced amino acid sequence of this novel protein contained 65 amino acid residues with 8 cysteine residues. Comparative sequence analysis showed that it was structurally related to cardiotoxin rather than neurotoxins. These results suggest that the venom of Bungarus multicinctus may contain cardiotoxin(s) which was not noticed before.

Amino Acid Sequence↗
PubMed1996

Testosterone enhances whereas estrogen inhibits calcium oxalate stone formation in ethylene glycol treated rats.

PURPOSE: The present study was conducted to extend our earlier study on the role of testosterone in the pathogenesis of urolithiasis and to further investigate the influence of sex hormone on the pathogenesis of calcium oxalate stone. MATERIALS AND METHODS: Adult Sprague-Dawley rats were divided into 9 groups, each containing 10 rats. Two groups of rats were left untreated and served as male and female controls. Another 7 groups of rats were fed a 0.5% ethylene glycol (EG) lithogenic diet for 4 weeks. Among them, 2 groups were male and female rats, 2 groups were castrated, 2 groups were castrated and then were subcutaneously implanted with testosterone, and 1 group of intact female rats was also subcutaneously implanted with testosterone. The stone and crystal deposits were examined by infrared spectrometer and polarizing and scanning electron microscope, respectively. Serum testosterone, creatinine and electrolytes and the renal excretion of, oxalate, citrate and creatinine were determined. RESULTS: Subcutaneous implantation of exogenous testosterone restored calcium oxalate stone formation in castrated, EG-treated male rats (80%) and enhanced urolithiasis in castrated female rats receiving EG (40%). However, the testosterone effect was less striking in intact female rats fed EG (10%). CONCLUSIONS: These data suggest that testosterone can promote and estrogen may inhibit calcium oxalate stone formation in EG-treated rats.

Animals↗
PubMed1996

Laparoscopic ligation of bilateral spermatic varices under epidural anesthesia.

Feasibility and safety of laparoscopic ligation of bilateral internal spermatic varices under epidural anesthesia were assessed in 11 patients. Another 11 patients undergoing the same procedure under ventilation-assisted anesthesia served as controls. Patients in both groups belonged to the American Society of Anesthesia functional class I. Arterial blood analyses were obtained (1) in the horizontal supine position; (2) in the 15 degrees Trendelenburg position; (3) at 15 min after carbon dioxide pneumoperitoneum insufflation, and (4) at 15 min after desufflation in the supine position. In the epidural anesthesia group arterial blood parameters and respiratory rate remained stable in the Trendelenburg position. After intraperitoneal insufflation of carbon dioxide for 15 min, the arterial carbon dioxide level increased from 40.1 +/- 2.2 to 42.1 +/- 2.6 mm Hg, the respiratory rate increased from 17.0 +/- 1.4 to 20.6 +/- 1.2/min, the blood pH value decreased from 7.386 +/- 0.027 to 7.355 +/- 0.034, all values showing statistically significant differences. These changes returned to the preinsufflation level 15 min after release of the pneumoperitoneum. The above-mentioned parameters remained unchanged under the pneumoperitoneum by assisted ventilation in the control group. The mean time of surgery was similar in both groups: 82 and 90 min for the groups having general and epidural anesthesia, respectively. All laparoscopic procedures were accomplished successfully under general anesthesia. However, failure to ligate the internal spermatic varices occurred in 3 patients under epidural anesthesia, mainly due to patient intolerance to abdominal distension. The operation was continued under intubated general anesthesia for relaxing the abdominal muscle to provide an adequate working space. In 8 patients being successfully operated under epidural anesthesia, 5 experienced mild but tolerable abdominal distension; 2 complained of shoulder pain intraoperatively. Although laparoscopic ligation of internal spermatic varices could be accomplished in some patients under epidural anesthesia, it carried a high failure rate, more intraoperative morbidity, and significant arterial blood gas alterations. Routine ventilation-assisted anesthesia is suggested for therapeutic laparoscopy even for an easy procedure such as the ligation of the internal spermatic varices.

Anesthesia, Epidural↗
PubMed1996

Adjuvant methotrexate, vinblastine and cisplatin chemotherapy for invasive transitional cell carcinoma: Taiwan experience.

PURPOSE: The feasibility of adjuvant cisplatin, methotrexate and vinblastine chemotherapy was evaluated in Taiwanese patients with invasive transitional cell carcinoma at high risk for recurrence. MATERIALS AND METHODS: We assigned 56 patients with high risk transitional cell carcinoma (vascular or lymphatic invasion in the primary tumor, poorly differentiated stage P2, P3, P4 or N+ and M0) to receive adjuvant chemotherapy after radical urological surgery. The chemotherapy consisted of 40 mg./m.2 methotrexate and 4 mg./m.2 vinblastine on days 1 and 8, and 100 mg./m.2 cisplatin on day 2 given in 6 courses at 21-day intervals. RESULTS: Median followup was 44 months. An average of 4.63 cycles of chemotherapy was administered. The median actual survival was 44 months, and the 1 and 3-year survival probabilities were 92% and 50%, respectively. The median disease-free survival was 15.5 months, and the 1 and 3-year disease-free survival probabilities were 66% and 28%, respectively. Only 5 (9%) and 1 (2%) patients had grades 3 and 4 leukopenia, respectively, and none died of sepsis. CONCLUSIONS: The use of adjuvant cisplatin, methotrexate and vinblastine chemotherapy in patients with invasive transitional cell carcinoma at high risk for recurrence is feasible with tolerable toxicity but randomized controlled trials will be required to assess the benefit.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1996

Dynamic infusion cavernosometry and cavernosography (DICC) in the evaluation of vasculogenic impotence.

BACKGROUND: This study was conducted to find more accurate physiological assessment of the penile vascular status in impotent patients. METHODS: Twenty-two males, aged 25-76 years (mean 54.0 years) with possible underlying vascular causes of impotence, had dynamic infusion cavernosometry and cavernosography (DICC) examination to ascertain the arterial and/or venous insufficiency in the penis. Prostaglandin E1 20 micrograms was injected intracavernously in this 4-phase study. RESULTS: No side effect or complication was noted during or after the examination in all 22 patients. During the phase 1 test, the mean resting intracavernous pressure (ICP) was 6.8 +/- 3.6 mmHg and the mean plateau ICP after PGE1 injection was 47.2 +/- 23.9 mmHg. The mean ratio of plateau ICP to mean arterial pressure was 50.2 +/- 23.4%. In the phase 2, the mean ICP determined 30 seconds after cessation of saline infusion without and with perineal compression was 65.3 +/- 26.8 mmHg and 129.1 +/- 26.2 mmHg, respectively. The mean ICP decay from 150 mmHg was 84.7 +/- 26.8 mmHg. The mean systolic pressure of each cavernous artery, 87.6 +/- 18.9 mmHg and 93.6 +/- 17.6 mmHg in left and right side, respectively, was found during phase 3 evaluation. The mean ratio of systolic pressure of each cavernous artery to brachial artery (cavernous-brachial index, CBI) was 70.1 +/- 12.7% and 75.1 +/- 11.5% in left and right side, respectively. Six patients with CBI greater than 75% had normal Doppler duplex sonography (diameter of cavernous artery greater than 0.9 mm and peak flow velocity greater than 25 cm/sec). Cavernosography in phase 4 evaluation showed venous leakage in 20 of 22 patients (90.9%) with ICP decay greater than 60 mmHg. Two patients with ICP decay less than 45 mmHg were found to have no venous leakage. CONCLUSIONS: The cavernous-brachial index is usually less than 75% in penile arterial insufficiency. The patients with ICP decay from 150 mmHg greater than 60 mmHg would have penile venous leakage demonstrated in the cavernosography. The results of cavernosometry are highly correlated with cavernosography. Our results suggest that DICC may provide a good examination for more physiological assessment and accurate diagnosis of vasculogenic impotence.

Adult↗
PubMed1996

Recurrence rate and risk factors for urinary calculi after extracorporeal shock wave lithotripsy.

PURPOSE: We determined the recurrence rate after extracorporeal shock wave lithotripsy (ESWL*) and evaluated its associated risk factors, such as patient sex and age, volume of daily water intake, related diseases, family history of urinary calculi, number, burden and location of stones, residual stones after ESWL, serum uric acid level before ESWL and so forth. MATERIALS AND METHODS: We conducted a retrospective study of 841 patients who underwent ESWL for primary treatment of urinary calculi between 1985 and 1987. Charts were reviewed for stone characteristics, outcomes and associated laboratory data. Followup 6 to 9 years later was done by mail and telephone interviews. RESULTS: Followup data were available for 436 patients (52%). Overall, stones recurred in 51% of the 436 patients at a mean followup of 7.1 years. Late recurrences were common, and were noted in as many as 70% of the patients after 9 years. This rate is greater than what has been previously reported after other treatments. Cox regression analysis indicated that the major risk factors associated with recurrence were male gender (relative risk 1.406, p < 0.05) and multiple stones (relative risk 1.777, p < 0.05). CONCLUSIONS: The frequent late recurrences support the speculation that ESWL might lead to greater recurrence rates. Therefore, the cost-effectiveness of ESWL versus other modalities for stone removal warrants more long-term evaluation.

Adolescent↗
PubMed1996

Structural determinants of cobrotoxin for binding to an enzyme-linked-immunoassay plate.

In order to assess the manner in which the structural state of a protein affects the results of an ELISA, the antigenic reactivities of cobrotoxin and reduced and S-carboxymethylated (RCM-) cobrotoxin with anti-RCM-cobrotoxin antibodies were studied. The results of competitive enzyme-linked immunoassay showed that the affinity of RCM-cobrotoxin for anti-RCM-cobrotoxin antibodies was higher than that of cobrotoxin. However, the cobrotoxin-coated wells had a greater reactivity towards anti-RCM-cobrotoxin antibodies than against RCM-cobrotoxin-coated wells. The lower reactivity observed with RCM-cobrotoxin-coated plates could be improved by adding 0.01% glutaraldehyde during the coating procedure. Studies on the antigenic structures of RCM-cobrotoxin showed that it contained an immunodominant epitope at residues 22-38. Moreover, the N-terminal and C-terminal regions of RCM-cobrotoxin encompassed other antigenic determinants which exhibited low reactivities towards anti-RCM-cobrotoxin antibodies. After removal of the antibodies against residues 22-38 of cobrotoxin from anti-RCM-cobrotoxin antibodies by passage through an affinity column, the remaining antibodies exhibited a similar reactivity towards cobrotoxin and RCM-cobrotoxin. The antibodies against residues 22-38 retained a little reactivity with the RCM-cobrotoxin-coated wells. These results suggest that the structural determinants of cobrotoxin and RCM-cobrotoxin for binding to the microtitre plates differ. Unlike RCM-cobrotoxin, the loop II structure of cobrotoxin encompassing residues 22-38 is not exclusively involved in the binding of cobrotoxin to microtitre plates.

Antibodies↗