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Biomedical subjects

L S Chang

Publications and source records attributed to L S Chang.

At least 127 records · Page 7Linked to original sources
PubMed1995

Chemical modification of cationic groups of a novel alpha-neurotoxin (Oh-4) from king cobra (Ophiophagus hannah) venom.

The cationic groups of arginine and lysine residues in Oh-4, a novel alpha-neurotoxin from king cobra (Ophiophagus hannah) venom were subjected to modification with p-hydroxyphenylglyoxal (HPG) and trinitrobenzene sulfonate (TNBS), respectively. Monoderivatization of Arg-35, resulted in a drastic loss in neurotoxicity to 25% of the native toxin. The activity was decreased to a greater extent with the derivative extensively modified on Arg-35, -9, and -37. The Arg-35-modified derivative retained about a half of the antigenicity of the native toxin, and extensive modification on Arg-9 and Arg-37 caused a further decrease in the antigenicity of the toxin molecule. Selective trinitrophenylation (TNP-) of Lys-51 caused losses of neurotoxicity and antigenicity by 77 and 83%, respectively. These results indicate that Arg-35 and Lys-51 in Oh-4 have important roles in the neurotoxicity. In contrast to the Arg residues at 9, 35, and 37, Lys-51 plays a more critical role in the antigenicity.

Amino Acids↗
PubMed1995

Characterization of epitopes in native and unfolded cobrotoxin: evidence of an immunodominant C-terminal region related to the production of precipitating and non-precipitating antibodies against cobrotoxin.

Rabbits hyperimmunized with cobrotoxin from Taiwan cobra venom produced non-precipitating as well as precipitating antibodies. Both antibody preparations exhibited higher affinity for native cobrotoxin than for reduced and S-carboxymethylated (RCM) cobrotoxin. This indicated that the epitope structures in cobrotoxin are mostly conformation-dependent. In order to identify the conformational epitopes, native cobrotoxin was hydrolyzed with acid protease A, and 12 peptides were obtained on HPLC. Three peptide fragments, AP-10, AP-11, and AP-12, showed pronounced antigenicities toward precipitating as well as non-precipitating antibodies. AP-10, AP-11, and AP-12 contained a common segment in the C-terminal region of cobrotoxin, residues 43 to 62, with intact disulfide linkages. Complete removal of the C-terminal antibodies from antisera and precipitating antibodies on a C-terminal segment-Sepharose affinity column resulted in the loss of their precipitability with cobrotoxin, whilst restoration of precipitability was observed on the addition of the C-terminal antibodies to the C-terminal antibody-depleted antisera and precipitating antibodies. Studies on the antigenic structures of RCM-cobrotoxin revealed that RCM-cobrotoxin contains an immunodominant epitope at positions 22-38. The N-terminal and C-terminal regions of RCM-cobrotoxin encompass other epitopes which exhibit low reactivities toward anti-RCM-cobrotoxin antibodies. However, no precipitated antigen-antibody complexes were observed with the mixture of anti-RCM-cobrotoxin antibodies and RCM-cobrotoxin. These results suggest that the inherently different immunogenicities with different segments might affect the precipitabilities of the resulting antibodies, and that the notable immunogenecity of the C-terminal region is related to the production of precipitating and non-precipitating antibodies against cobrotoxin.

Amino Acid Sequence↗
PubMed1995

Role of amino and carboxyl groups of cobrotoxin in the conformational stability and the interaction with acetylcholine receptor.

To study the functional involvements of the common interaction of the Leu-1 alpha-amino group and Asp-58 in cobrotoxin, the lysine epsilon-amino groups of cobrotoxin were initially guanidinated with o-methylisourea. The alpha-amino group of Leu-1 was them modified with TNBS after the guanidination of cobrotoxin. Both modified derivatives displayed no significant changes in the secondary structure and antigenicity of cobrotoxin, whereas the binding affinity for nicotinic acetylcholine receptor (nAChR) was pronouncedly decreased when Leu-1 was modified. Six out of seven free carboxyl groups and the remaining buried Glu-21 carboxyl group of cobrotoxin were modified with glycine methyl ester in the absence and presence of guanidine HCl, respectively. Alternation in the beta-sheet secondary structure of cobrotoxin was observed with the carboxyl-group modified derivatives, which caused a decrease in the binding activity of the toxin molecule to the antibody and nAChR. Moreover, modification of the Glu-21 carboxyl group of cobrotoxin further reduced the nAChR binding activity, while the antigenicity remained unchange. Thus, our results conclude that the Glu-21 residue and the common interaction of the terminal Leu-1 alpha-amino group and the Asp-58 carboxyl group are related to the nAChR-binding activity of cobrotoxin, and the free carboxyl groups in cobrotoxin are conformation-essential.

Amines↗
PubMed1995

Differential roles of two tandem E2F sites in repression of the human p107 promoter by retinoblastoma and p107 proteins.

Although many lines of evidence indicate that the cellular protein p107 is closely related to the retinoblastoma protein, the exact function of the p107 gene and its regulation are presently not known. To investigate the molecular mechanism controlling expression of the human p107 gene, a 5' flanking sequence of this gene was isolated and shown to promote high-level expression of a luciferase reporter gene in cycling human 293 and Saos-2 cells. Sequencing and transcription mapping analyses showed that the human p107 promoter is TATA-less and contains a tandem, direct repeat of E2F-binding sites, with the 3' copy overlapping the major transcription initiation site. Deletion analysis of the p107 promoter showed that a promoter DNA fragment containing only the two E2F sites together with the leader sequence could direct relatively efficient expression in 293 cells. Site-directed mutagenesis of these E2F sites revealed that although both sites were important for p107 promoter activity, mutation on the proximal, initiation site copy of the E2F site showed a stronger effect. The human p107 promoter could be repressed by the retinoblastoma protein and its own gene product. Interestingly, the repression was found to be mediated through the 5' copy of the E2F site. These studies demonstrate for the first time differential roles of two tandem E2F sites in promoter regulation.

Base Sequence↗
PubMed1995

Evidence showing a different repertoire of antibodies against unfolded cobrotoxin in anticobrotoxin and anti-reduced and S-carboxymethylated cobrotoxin antisera.

Antibodies which specifically reacted with reduced and S-carboxy-methylated(RCM) cobrotoxin were purified from anticobrotoxin antisera and anti-RCM-cobrotoxin antisera, respectively. Results using a competitive immunoassay revealed that the antibodies from anti-RCM-cobrotoxin antisera had a greater affinity for RCM-cobrotoxin than for cobrotoxin. Whereas the reactivity of the antibodies from the anticobrotoxin antisera toward cobrotoxin and RCM-cobrotoxin had a reversed order of binding. In contrast to observations made with S. aureus V8 protease-digest hydrolysates, the antigenic structures of RCM-cobrotoxin recognized by antibodies from anti-RCM-cobrotoxin antisera were notably affected following hydrolysis of RCM-cobrotoxin with chymotrypsin. Moreover, the chymotryptic hydrolysates showed a comparable reactivity as RCM-cobrotoxin toward the antibodies purified from anticobrotoxin antibodies, but a decrease in antigenicity with the V8 protease hydrolysates. These results reveal that the repertoire of antibodies against the unfolded cobrotoxin are not the same in anticobrotoxin and anti-RCM-cobrotoxin antisera. Moreover, it suggests that the repertoire of antibodies from the different sources against the same antigen can be differentiated by measurement of their reactivities with the proteolytic hydrolysates of the antigen.

Animals↗
PubMed1995

The impact of pneumoperitoneum, pneumoretroperitoneum, and gasless laparoscopy on the systemic and renal hemodynamics.

BACKGROUND: The use of therapeutic laparoscopy has become widespread recently. It is important to assess the effects of gaseous and gasless laparoscopy on systemic and renal hemodynamics. STUDY DESIGN: A prospective controlled animal study was performed on 40 well-hydrated pigs. Systemic and renal hemodynamics were monitored during peritoneal insufflation, retroperitoneal insufflation, and abdominal wall lifting for a period of two hours. A laser Doppler flow meter was applied laparoscopically to measure the renal cortical tissue perfusion. RESULTS: Peritoneal insufflation of carbon dioxide to a pressure of 15 mm Hg elicited transient elevations of the aortic pressure and carotid arterial blood flow. Unilateral pneumoretroperitoneum caused a smaller change on systemic hemodynamics. Pneumoperitoneum and pneumoretroperitoneum caused oliguria. Superficial renal cortical blood flow reduction decreased by an average of 60 percent in the compressed kidney, and blood flow returned to the pre-insufflation level after the pressure was released. A gradual decrease of tissue perfusion in the contralateral kidney and a concomitant gradual increase of the intra-abdominal pressure were observed when pneumo-retroperitoneum was maintained for two hours. No significant changes in urinary output and in systemic and renal hemodynamics were found when the abdominal wall was lifted up with a force equivalent to 15 mm Hg. CONCLUSIONS: Significant systemic and renal hemodynamic changes were elicited in gaseous but not in gasless laparoscopy, which may explain the decreased urinary output observed during gaseous laparoscopy. Pneumoperitoneum caused greater systemic and renal hemodynamic alterations than pneumoretroperitoneum; however, the effects were transient and reversible after a period of two hours.

Animals↗
PubMed1995

Metabolic and behavioral response in rats treated with amphetamine chronically with and without challenge.

Chronic amphetamine (AMPH) treatment may cause behavioral sensitization in animals and can be used as an animal model of psychosis. The aim of the study was to check the behavioral and metabolic response in this animal model. In rats pretreated with normal saline (NS) or AMPH, with or without AMPH challenge, the [14C]deoxyglucose method was employed to check the metabolic changes in 42 regions. Behavioral testing was performed in rats with the same treatment. The results showed that after challenge with AMPH, glucose utilization was enhanced in most of the regions investigated. However, metabolic enhancement of the AMPH-pretreated group was lower in the caudate nucleus when compared with that of the NS-pretreated group though the stereotypy rating was higher in the former. Dissociation between the metabolic enhancement and behavioral response was noted. Furthermore, more significant differences between the two pretreated conditions of glucose utilization were found with challenge than without challenge. Further evaluation using procedures which include advanced techniques can be applied in further investigation.

Amphetamine↗
PubMed1995

Does prediction of epitopes from the primary structure of a protein represent the epitope in the native structure? A study using cobrotoxin.

In contrast to observations made with S. aureus V8 protease-digest hydrolysates, the antigenic structures of reduced and S-carboxymethylated (RCM)-cobrotixin were notably affected following hydrolysis of RCM-cobrotoxin with chymotrypsin. The peptide separated from the V8 protease-digest hydrolysates with a sequence at positions 22-38 of cobrotoxin exhibited a nearly equal reactivity toward the anti-RCM-cobrotoxin antibodies as RCM-cobrotoxin. Chymotryptic cleavage on this segment caused a precipitous drop in the antigenicity of RCM-cobrotoxin. Alternatively, the N-terminal and C-terminal regions of RCM-cobrotoxin encompassed other antigenic determinants which exhibited low reactivities toward anti-RCM-cobrotoxin antibodies. The epitope structures of RCM-cobrotoxin are in line with those predicted from the hydrophobicity profile of cobrotoxin, but the notably immunoreactive region in the C-terminal region of native toxin molecule (Ref. 1) cannot be predicted from analysis of its primary structure. Moreover, RCM-cobrotoxin had a superior reactivity toward anti-RCM-cobrotoxin antibodies than cobrotoxin did. These results indicate that the epitope structures in RCM-cobrotoxin and cobrotoxin are different.

Antibodies↗
PubMed1995

The subjective assessment and pressure-flow study of outcome of surgical treatment for patients with prostatism and high voiding pressure.

BACKGROUND: In 1992, the International Continence Society (ICS) began to conduct an international multicenter study to correlate the symptoms and urodynamics in patients with BPH. Our institute was invited to participate this ICS-BPH study, and the preliminary (phase I) results have revealed poor correlation between preoperative symptoms and parameters of the pressure-flow study. This phase II study continued the phase I ICS-BPH study to assess the changes of subjective symptoms and objective parameters of pressure-flow study before and after TURP. METHODS: Fifty-two patients with prostatism, aged from 56 to 80, were evaluated using the ICS-BPH questionnaire, free urine flow rate and pressure-flow study. According to the criteria of maximum urine flow rate (Qmax) 15 ml/sec and voiding intravesical pressure at Qmax (Pves at Qmax) 100 cm H2O, low flow-high pressure (LFHP) was proved in 20 patients (38%), high flow-high pressure (HFHP) in 22 (42%), low flow-low pressure (LFLP) in 3 (6%), and high flow-low pressure (HFLP) in 7 (14%). Transurethral resection of the prostate (TURP) was performed for 33 of 42 patients with high voiding pressure (LFHP and HFHP groups). RESULTS: The weight of the TURP specimen was 28 +/- 6.3 gm. Sixteen patients repeated the questionnaire and pressure-flow study 9.0 +/- 2.1 months after surgery. Both irritative and obstructive symptom scores decreased in 15 (94%) of the 16 patients (p < 0.001). The opening intravesical pressure (openPves) and Pves at Qmax improved from 136 +/- 34 and 124 +/- 30 cm H2O preoperatively to 89 +/- 25 and 94 +/- 22 cm H2O postoperatively respectively (p < 0.004). However, UFRmax, 16.7 +/- 6.0 ml/sec preoperatively and 19.3 +/- 6.4 postoperatively, showed no statistical difference. If the HFHP and LFHP groups were compared separately, the degree of improvement in symptomatology or parameters of pressure-flow study was similar, although changes in HFHP group appeared more evident. UFRmax in LFHP group changed significantly from 11.9 +/- 2.8 preoperatively to 22.1 +/- 6.7 ml/sec postoperatively. For the patients with high flow obstruction (HFHP group), although UFRmax did not increase after TURP, the symptom score, openPves and Pves at Qmax, decreased as much as the LFHP group did. CONCLUSIONS: Based on the analysis of the pressure flow study, TURP not only offers an advantage to the patient with classical obstruction (LFHP group), but also to patients with high flow obstruction (HFHP group). However, this does not suggest that a pressure-flow study should be performed routinely for a male with prostatism. It is advised only for patients with Qmax > 15 ml/sec but exhibiting typical symptoms.

Aged↗
PubMed1994

Hb E/Hb LeporeHollandia in a family from Bangladesh.

We describe a family from Bangladesh in which three children are compound heterozygotes for Hb E (alpha 2 beta 2, beta 26Glu Lys) and Hb Lepore (delta-beta fusion gene). PCR amplification and direct nucleotide sequencing established that the fusion gene is Hb LeporeHollandia, with the cross-over localized to a 40 bp window between codon 22 and IVS-1 nt 16 of the delta- and beta-globin genes. This unusual combination of mutations is associated with a relatively mild clinical phenotype, with all three affected siblings having microcytic anemia of moderate severity without the need for transfusions.

Bangladesh↗
PubMed1994

Energy transfer from tryptophan residues of proteins to 8-anilinonaphthalene-1-sulfonate.

The binding of the apolar fluorescent dye 8-anilinonaphthalene-1-sulfonate (ANS) to bovine serum albumin (BSA), phospholipase A2 (PLA2), ovalbumin, lysozyme, cobrotoxin and N-acetyltryptophanamide was used to assess the factors affecting the efficiency of energy transfer from Trp residues to the ANS molecule. We found that the efficiency of energy transfer from Trp residues to ANS was associated with the ability of proteins to enhance the ANS fluorescence. At the same molar concentration of protein, BSA enhanced ANS fluorescence most among these proteins; its Trp fluorescence was drastically quenched by the addition of ANS. Fluorescence enhancement of ANS in PLA2-ANS complex increased upon addition of Ca2+ or change of the buffer to acidic pH, resulting in a higher efficiency of energy transfer from Trp residues to ANS. There was limited ANS fluorescence enhancement with ovalbumin, lysozyme, cobrotoxin, and N-acetyltryptophanamide and a less efficient quenching in Trp fluorescence. The capabilities of proteins for binding with ANS correlated with the decrease in their Trp fluorescence being quenching by ANS. However, the microenvironment surrounding Trp residues of proteins did not affect the energy transfer. Based on these results, the factors that affected the energy transfer from Trp residues to ANS are discussed.

Anilino Naphthalenesulfonates↗
PubMed1994

Functional involvement of Lys-6 in the enzymatic activity of phospholipase A2 from Bungarus multicinctus (Taiwan banded krait) snake venom.

Phospholipase A2 (PLA2) from Bungarus multicinctus snake venom was subjected to Lys modification with 4-chloro-3,5-dinitrobenzoate and trinitrobenzene sulfonic acid, and one major carboxydinitrophenylated (CDNP) PLA2 and two trinitrophenylated (TNP) derivatives (TNP-1 and TNP-2) were separated by high-performance liquid chromatography. The results of amino acid analysis and sequence determination revealed that CDNP-PLA2 and TNP-1 contained one modified Lys residue at position 6, and both Lys-6 and Lys-62 were modified in TNP-2. It seemed that the Lys-6 was more accessible to modified reagents than other Lys residues in PLA2. Modification of Lys-6 caused a 94% drop in enzymatic activity as observed with CDNP-PLA2 and TNP-1. Alternatively, the enzyme modified on both Lys-6 and Lys-62 retained little PLA2 activity. Either carboxydinitrophenylation or trinitrophenylation did not significantly affect the secondary structure of the enzyme molecule as revealed by the CD spectra, and Ca2+ binding and antigenicity of Lys-6-modified PLA2 were unaffected. Conversion of nitro groups to amino groups resulted in a partial restoration of enzymatic activity of CDNP-PLA2 to 32% of that of PLA2. It reflected that the positively charged side chain of Lys-6 might play an exclusive role in PLA2 activity. The TNP derivatives could be regenerated with hydrazine hydrochloride. The biological activity of the regenerated PLA2 is almost the same as that of native PLA2. These results suggest that the intact Lys-6 is essential for the enzymatic activity of PLA2, and that incorporation of a bulky CDNP or TNP group on Lys-6 might give rise to the distortion of the interaction between substrate and the enzyme molecule, and the active conformation of PLA2.

Amino Acid Sequence↗
PubMed1994

The essentiality of B chain in stabilizing the structure of the A chain in beta 1-bungarotoxin from Bungarus multicinctus venom.

The dynamic of Trp residue in beta 1-bungarotoxin (beta 1-Bgt), the A chain of beta 1-Bgt and phospholipase A2 (PLA2) was assessed by fluorescence measurement. Acrylamide quenching studies showed that the exposure degree of the Trp in PLA2 is higher than the Trp in beta 1-Bgt. The Trp of beta 1-Bgt had a higher accessibility for iodide, reflecting that the basic nature of the B chain might exert an attractive electrostatic force for iodide and increase the susceptibility of Trp in the A chain to iodide. Removal of the B chain of beta 1-Bgt did not significantly affect the exposure degree of Trp in the A chain. Alternatively, the polarity of the environment around the Trp and the hydrophobic character of ANS and substrate binding sites in the separated A chain changed. Measurement of Trp fluorescence with increasing temperature showed that the stability of structure of beta 1-Bgt was higher than those of the separated A chain and PLA2. These results suggest that the B chain might interact with the A chain and stabilize the conformation of the A chain in beta 1-Bgt.

Animals↗
PubMed1994

The time for serum testosterone to reach castrate level after bilateral orchiectomy or oral estrogen in the management of metastatic prostatic cancer.

OBJECTIVE: To understand the time for serum testosterone to reach castrate level after bilateral orchiectomy or oral estrogen in the management of metastatic prostatic cancer. METHODS: A total of 20 consecutive patients with adenocarcinoma of the prostate with bony metastasis were enrolled in this study. Their mean age was 72.8 years old (range, 57 to 82 years). Pretreatment serum testosterone levels were obtained in all men. Thirteen men were treated with bilateral orchiectomy. Immediately after removal of testes, serial blood samplings for serum testosterone levels were drawn every fifteen minutes for the first two hours, then hourly for another sixteen hours. Seven men were treated with oral estrogen diethylstilbestrol (DES), 3 mg per day. Serum testosterone levels were checked on a weekly basis for two months, and then biweekly for another two months. RESULTS: Castration time of bilateral orchiectomy ranges from three to twelve hours (mean, 8.6 hours). The biological half-life of serum testosterone was from thirty to sixty minutes (mean, 45 minutes). Castration time of oral estrogen (DES) was from twenty-one to sixty days (mean, 38.3 days). CONCLUSIONS: Bilateral orchiectomy and oral estrogen were both effective ways of castration for patients with bony metastatic prostatic cancer. Bilateral orchiectomy provides a more rapid castration and is one hundred seven times faster than oral estrogen in reaching castrate level.

Adenocarcinoma↗
PubMed1994

The long-term stone recurrence rate and renal function change in unilateral nephrectomy urolithiasis patients.

We evaluated the stone recurrence rate and long-term renal function in 50 patients with a solitary kidney 60 to 84 months (mean 70.6) after unilateral nephrectomy for urinary tract stone disease. Followup evaluation included a detailed history, physical examination, blood and urine biochemistry studies, urinalysis, urine culture, excretory urography, ultrasonography of the kidney and 131iodine-ortho-iodohippurate renography. The overall stone recurrence rate in unilateral nephrectomy urolithiasis patients was 30% (15 of 50). The mean interval until stone recurrence was 31.1 months (range 6 to 74) and the mean episodes of recurrence were 2.1 times per patient (range 1 to 5). Function of the remnant kidney in most patients was unchanged during followup. However, 2 of the 15 patients with recurrent stones had anuria during the acute attack of renal colic and, thus, required percutaneous nephrostomy urinary diversion, while 1 had proteinuria (3 gm. per day) and progressive renal failure 47 months after nephrectomy. The metabolic stone patients seemed to experience recurrence more easily than metabolic stone patients seemed to experience recurrence more easily than infection stone patients (37% versus 13%) but no statistically significant difference was noted (p = 0.198).

Adult↗
PubMed1994

Immunological neutralization of cobrotoxin by its homologous precipitin and non-precipitin antibodies.

Anti-cobrotoxin antibodies can be separated into precipitin and non-precipitin antibodies. The precipitin antibody possesses the same binding affinity to cobrotoxin as non-precipitin antibody, but the neutralizing capability of the latter is superior to that of the former in blocking cobrotoxin binding to nicotinic acetylcholine receptor (nAChR). After preincubation with antibodies, cobrotoxin completely lost its binding activity to nAChR. The dissociation of cobrotoxin-nAChR complex by the antibodies was low, and 60% of the complex formation appeared to be irreversible. These results indicate that the neutralization of cobrotoxin by the antibody may predominantly involve unbound, receptor-free cobrotoxin. The relationships of neutralization capacity and binding affinity as well as bond strength between cobrotoxin and its antibodies are incongruous. Different local conformational changes of a unique Trp in cobrotoxin on binding with the precipitin and non-precipitin antibodies seem to lead to different accessibility for fluorescence quenchers. Characterization of the binding domains by immunoprecipitation with the antibodies correlated with the quenching results. Thus, the binding topography of cobrotoxin may play an important role over the binding affinity and bond strength in neutralization by cobrotoxin antibody.

Animals↗
PubMed1994

Morphological change in the urothelium after electrohydraulic versus pulsed dye laser lithotripsy.

OBJECTIVE: To observe morphological changes in the urothelium after exposure to laser beam and electrohydraulic spark. MATERIALS AND METHODS: The pulse dye laser lithotripter (MDL-2, Candela Corporation, Boston, USA) and the electrohydraulic lithotripter (AEH-2, Circon ACMI Corporation Stamford, USA) were used directly on the bladder mucosa in six pigs. Immediately after treatment, the bladder mucosa was prepared for light microscopic and scanning electron microscopic examination. RESULTS: Both electrohydraulic spark and laser beam caused mucosal denudation, submucosal swelling and haemorrhage. There was a positive correlation between the degree of urothelial damage and the power and pulse numbers applied. CONCLUSION: Direct contact of the urothelium in patients treated with laser beam or electrohydraulic spark should be avoided.

Animals↗