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Biomedical subjects

S J Yang

Publications and source records attributed to S J Yang.

At least 19 recordsLinked to original sources
PubMed1996

Tirapazamine (SR 4233) interrupts cell cycle progression and induces apoptosis.

Tirapazamine (Tira), a bioreductive agent, is highly toxic to cells under low oxygen conditions. Since active investigations of this agent are focusing on its potential as an adjunct of radiotherapy to improve overall effects on radioresistant hypoxic tumor cells, understanding its toxic mechanisms under aerobic conditions is important to the clinical application of this agent. Tira-treated V79 Chinese hamster cells were tested for cytotoxicity by colony assay and growth inhibition by the MTT assay. The survival of V79 cells after being exposed to 100 microM of Tira for 2 h was about 78% of untreated controls. The mitotic cell counts of V79 cells approached zero after 4 h treatment of Tira at 100 microM or 3 h at 300 microM. The fragmentation pattern of DNA isolated from cells 2 h after 300 microM Tira treatment showed characteristics of apoptotic cells. The induction of apoptosis by Tira was also detected by flow cytometric analysis and microscopic observation. These effects of Tira may be part of underlying toxic mechanisms to cells (including normal cells) under aerobic conditions.

Animals↗
PubMed1996

Subunit structure of vacuolar proton-pyrophosphatase as determined by radiation inactivation.

Vacuolar proton-pyrophosphatase (H(+)-PPase) of mung bean seedlings contains a single kind of polypeptide with a molecular mass of approx. 73 kDa. However, in this study, a molecular mass of approx. 140 kDa was obtained for the purified vacuolar H(+)-PPase by size-exclusion gel-filtration chromatography, suggesting that the solubilized form of this enzyme is a dimer. Radiation inactivation analysis of tonoplast vesicles yielded functional masses of 141.5 +/- 10.8 and 158.4 +/- 19.5 kDa for PP1 hydrolysis activity and its supported proton translocation respectively. These results confirmed the in situ dimeric structure of the membrane-bound H(+)-PPase of plant vacuoles. Further target-size analysis showed that the functional unit of purified vacuolar H(+)-PPase was 71.1 +/- 6.7 kDa, indicating that only one subunit of the purified dimeric complex would sufficiently display its enzymic reaction. Moreover, in the presence of valinomycin and KCl, the functional size of membrane-bound H(+)-PPase was decreased to approx. 63.4 +/- 6.3 kDa. A working model was proposed to elucidate the structure of native H(+)-PPase on vacuolar membrane as a functional dimer. Factors that would disturb the membrane, e.g. membrane solubilization and the addition of valinomycin and KCl, may induce an alteration in its enzyme structure, subsequently resulting in a different functional size.

Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone↗
PubMed1996

Involvement of tyrosine residue in the inhibition of plant vacuolar H(+)-pyrophosphatase by tetranitromethane.

Plant vacuolar vesicles contain a novel H(+)-translocating pyrophosphatase (H(+)-PPase, EC 3.6.1.1). Modification of tonoplast vesicles and purified vacuolar H(+)-PPase from etiolated mung bean seedlings with tetranitromethane (TNM) resulted in a progressive decline in H(+)-translocating pyrophosphatase activity. The half-maximal inhibition was brought about by 0.6, 1.0, and 0.8 mM TNM for purified and membrane-bound H(+)-PPases, and its associated proton translocation, respectively. The maximal inhibition of vacuolar H(+)-PPase by TNM occurred at a pH value above 8. Loss of activity of purified H(+)-pyrophosphatase followed pseudo-first order rate kinetics, yielding a first-order rate constant (k2) of 0.039 s(-1) and a steady-state dissociation constant of inactivation (Ki) of 0.02 mM. Covalent modification of vacuolar H(+)-PPase by TNM increased Km value of the enzyme for its substrate without a significant effect on Vmax. Double logarithmic plots of the pseudo-first order rate constant (kobs) versus TNM concentration exhibited a slope of 0.88, suggesting that at least one tyrosine residue was involved in the inactivation of H(+)-PPase enzymatic activity. Further spectrophotometric measurements of the nitrated H(+)-pyrophosphatase indicated that TNM could modify approximately two tyrosine residues/subunit of the enzyme. However, Tsou's analysis revealed that only one of those modified tyrosine residues directly participated in the inhibition of enzymatic activity of vacuolar H(+)-PPase. The physiological substrate, i.e., dimagnesium pyrophosphate, provided substantial protection against inactivation by TNM. Moreover, NEM pretreatment of the enzyme decreased the number of subsequent nitration of vacuolar H(+)-PPase. Taken together, we suggest that vacuolar H(+)-pyrophosphatase contains a substrate-protectable tyrosine residue conferring to the inhibition of its activity and this tyrosine residue may be located in a domain sensitive to the modification of Cys-634 by NEM.

Electrophoresis, Polyacrylamide Gel↗
PubMed1996

Group-specific identification of polioviruses by PCR using primers containing mixed-base or deoxyinosine residue at positions of codon degeneracy.

We have developed a method for differentiating polioviruses from nonpolio enteroviruses using PCR. A pair of panpoliovirus PCR primers were designed to match intervals encoding amino acid sequences within VP1 that are strongly conserved among polioviruses. The initiating primer hybridizes with codons of a 7-amino-acid sequence that has been found only in polioviruses; the second primer matches codons of a domain thought to interact with the cell receptor. The panpoliovirus PCR primers contain mixed-base and deoxyinosine residues to compensate for the high degeneracy of the targeted codons. All RNAs from 48 vaccine-related and 110 wild poliovirus isolates of all three serotypes served as efficient templates for amplification of 79-bp product. None of the genomic sequences of 49 nonpolio enterovirus reference strains were amplified under equivalent reaction conditions. Sensitivities of poliovirus detection were as low as 100 fg (equivalent to approximately 25,000 genomic copies or 25 to 250 PFU) when the amplified products were visualized by ethidium bromide fluorescence. These degenerate PCR primers should aid in the detection of all polioviruses, including those wild poliovirus isolates for which genotype-specific reagents are unavailable.

Amino Acid Sequence↗
PubMed1996

Effects of tetrandrine on cytosolic free calcium in cultured rat myocardial cells.

AIM: To study the effects of tetrandrine (Tet) on myocardium. METHODS: Using Fura 2-AM and AR-CM-MIC cation measurement system, cytosolic free calcium ([Ca2+]i) was examined in cultured rat single myocardial cells. RESULTS: The resting [Ca2+]i was 90 +/- 12 nmol.L-1 in the presence of Ca2+ 1.3 mmol.L-1 in Hanks' solution. Tet 1-100 mumol.L-1 had no effect on the resting [Ca2+]i, but 10-100 mumol.L-1 depressed the [Ca2+]i elevation when extracellular Ca2+ was 5 mmol.L-1. Tet 1-100 mumol.L-1 inhibited KC1 (30 and 60 mmol.L-1) induced [Ca2+]i elevation in a concentration-dependent manner, the IC50 value was 8.8 mumol.L-1 (95% confidence limits: 3.3-23.7 mumol.L-1) and 6.9 mumol.L-1 (95% confidence limits: 2.8-17.4 mumol.L-1), respectively. Norepinephrine (NE) 10 mumol.L-1 caused a rapid increase in [Ca2+]i in the presence or absence of extracellular Ca2+, Tet 30-100 mumol.L-1 only decreased the former. Tet 10-100 mumol.L-1 also decreased ouabain (Oua)-induced elevation in [Ca2+]i. CONCLUSION: Tet had inhibitory effects on Ca2+ transmembrane movement, but it is not a selective calcium channel blocker in rat myocardial cells.

Alkaloids↗
PubMed1995

Demonstration of the presence of protease-cutting site in the spacer of hepatitis B viral Pol protein.

Molecular genetic studies have revealed that the human hepatitis B viral (HBV) Pol protein, a polypeptide of about 94 kDa, contains four domains. These are the 5'-terminal protein, spacer, RNA reverse transcriptase/DNA polymerase, and RNase H, respectively, from the amino (N) to carboxy (C) terminus. No evidence indicates as yet the involvement of a specific protease in cleaving the Pol protein or the presence of protease-cutting sites in the Pol protein. An in vitro-translated Pol protein was shown to be cleaved by purified thrombin but not in the presence of its inhibitor, hirudin. Two thrombin-cutting sites, spanning 194 amino acids, were then deduced by thrombin digestion of Pol protein with various lengths of C-terminal deletion. These two putative cutting sites, one located in the spacer region and the other in the beginning of the polymerase region, were found to be conserved at similar positions in the Pol protein of all hepadnaviruses. By using a novel method called the LacZ localization assay (LLA), it was demonstrated that a tripartite fusion protein containing the nucleus localization sequence (NLS) of SV40 large T Ag, the putative thrombin cutting sequence (Ile-Arg-Ile-Pro-Arg320-Thr) of HBV Pol protein and the full length beta-galactosidase of E. coli, exhibited a lower percentage (approximately 53%) of targeting into the nucleus of transfected hepatoma cells when compared with a similar tripartite protein containing a single mutation (Arg320 residue into Trp320) of HBV Pol protein (approximately 78%) or with a bipartite protein of SV40 NLS-beta-galactosidase (approximately 90%). These results indicate that the putative thrombin-cutting site in the spacer region of HBV Pol protein could be cleaved by a cellular protease resulting in the separation of NLS sequence from the beta-galactosidase and rendering a lower frequency of X-gal staining in the nucleus.

Amino Acid Sequence↗
PubMed1995

[Influence of the 2nd and 3rd grade abstracts from scorpion venom of Buthus martensii Karsch on action potentials of fast response myocardiocytes].

AIM: To screen the active components of sodium channel blockade action from Buthus martensii Karsch (BMK) scorpion venom. METHODS: Myocardiocytes of mice were cultured. Action potentials of fast response myocardiocytes were recorded. The effects of 22 contents of BMK scorpion venom in a concentration of 3 mg.L-1 were tested and compared with that of TTX 2.5 mg.L-1, nimodipine (Nim) 3 mg.L-1 and BaCl2 24.4 mg.L-1. RESULTS: BMK-1 and other 19 contents significantly decreased depolarization parameters Vmax, APA, OS, and MDP, which was similar to that of TTX and different from that of Nim and BaCl2. CONCLUSION: The variations in the action potentials of fast response myocardiolcytes indicated that 20 contents of BMK scorpion venom had sodium channel blockade action.

Action Potentials↗
PubMed1995

[Calcium channel blockade and anti-free-radical actions of panaxadiol saponins Rb1, Rb2, Rb3, Rc, and Rd].

AIM: To identify the calcium channel blockade and anti-free-radical actions of panaxadiol saponins Rb1, Rb2, Rb3, Rc, and Rd. METHODS: On ventricular myocardiocytes of Wistar rats, single channel activities of L, T, and B type Ca2+ channels were recorded with the cell-attached configuration of patch-clamp technic, and free radical contents were measured with electron spin resenance method. RESULTS: Rb1, Rb2, Rb3, and Rc 200 mumol.L-1 shortened the open times, prolonged the close times, and reduced the openstate probabilities of calcium channels and 30 mumol.L-1 antagonized the increase of free radical content induced by xanthine 0.42 mmol.L-1-xanthine oxidase 5.3 nmol.L-1, but Rd in the same dose behaved none of the effects. CONCLUSION: Rb1, Rb2, Rb3, and Rc had both the calcium channel blockade and anti-free-radical actions.

Animals↗
PubMed1995

[Effects of tetrahydroprotoberberines on cytosolic free calcium in cultured rat single myocardial cells].

Effect of tetrahydroberberine (THB), l-tetrahydropalmatine (THP) and l-stepholidine (SPD) were supposed to be related to the blocking of calcium influx. In this paper, using Fura-2/AM and AR-CM-MIC cation measurement system, the effects of THB, THP and SPD on cytosolic free calcium ([Ca2+]i) in cultured rat single myocardial cells were examined and compared with verapamil (Ver). THB, THP and SPD (10-100 mumol.L-1) were found to increase resting [Ca2+]i gently, which was not depressed by tetrodotoxin. THB, THP and SPD (1-100 mumol.L-1) were also shown to inhibit the KCl-induced [Ca2+]i elevation, the IC50 values of THB and SPD were 50.9 (95% confidence limits: 18.5-140) mumol.L-1 and 23.5 (95% confidence limits 7.6-73.4) mumol.L-1, respectively. THB, THP and SPD 30 mumol.L-1 were also shown to inhibit the elevation of [Ca2+]i induced by high extracellular calcium and norepinephrine; but the inhibitory effects of these drugs were weaker than those of Ver. The three compounds showed no significant effect on ouabain induced [Ca2+]i increase. These results suggest that the inhibitory effects of THB, THP and SPD on [Ca2+]i in myocite by blocking voltage-dependent calcium channels were similar but inferior to Ver.

Animals↗
PubMed1994

[Anti-hypoxia and anti-glucose deficiency effects of PQS on guinea pig papillary muscle in low frequency electrical stimulation].

Panax quinquefolium saponin (PQS) increases the contractibility of papillary muscle of guinea pigs during electrical stimulation with frequency of 1/8-1/64Hz (P < 0.05). In Krebs-Henseleit solution purged with gas containing 95% N2 and 5% CO2 regardless of containing glucose or not, PQS (0.3mg/kg) has the effect of anti-hypoxia and anti-glucose deficiency (P < 0.05, P < 0.01) in the electrical stimulation with frequency of 1/8-1/64Hz.

Animals↗
PubMed1994

[The effects of Panax quinquefolium saponin (PQS) and its monomer ginsenoside on heart].

Experiments have shown that PQS (0.03-3 mg/ml) can inhibit the contractility of papillary muscle of guinea pigs, and on depolarized sample of papillary muscle with high potassium, PQS (0.03-0.3 mg/ml) can increase this contractility. Monomer saponin-Re (10 mg/kg),-Rb3 (30 mg/kg) can inhibit the hemodynamic indication of rats, but pseudogisenoside-F11 (10 mg/kg) acts the other way round. These results prove that PQS contains two components of opposite actions.

Animals↗
PubMed1994

Characteristics of immunocomplex in autopsy tissues of hemorrhagic fever with renal syndrome.

Using repeated PAP or repeated PAP and ABC immunocytochemical methods, we were able to demonstrate viral antigens, Ig and Clq in the tissues of 20 autopsy materials that had been preserved for 3-30 years. Serial paraffin sections were stained with the first antibodies against both viral antigens (G2 and Np) and human IgG, IgM, C3 and Clq. Immunocomplex, composed of viral antigen, IgG and Clq were found diffusely in the cells of each organ and extracellularly in the sera, various secretions and exudates. When stained by A25 etc, coarse granular antigen or inclusion bodies were found without demonstrable Ig and Clq. It was concluded that the immunocomplex in tissues of epidemic hemorrhagic fever in Shaanxi Province, China was both intracellular and extracellular and was perhaps soluble due to antigens in excess, with characteristics quite different from that of other immune diseases.

Animals↗
PubMed1992

Genotype-specific in vitro amplification of sequences of the wild type 3 polioviruses from Mexico and Guatemala.

The extensive nucleotide sequence heterogeneity among independent genotypes of wild polioviruses permits the systematic design of genotype-specific molecular reagents. We have prepared two sets of polymerase chain reaction (PCR) primer pairs specific for the genotype of wild poliovirus type 3 recently endemic to Mexico and Guatemala. Nucleotide sequences of a representative wild type 3 virus isolated in Mexico in 1989 differed from the corresponding Sabin 3 (Leon 12 a1b) sequences at 167 of 900 positions within the VP1 region. From the sequence data, wild virus-specific primer pairs were designed to complement regions of high mismatch (greater than 33%) with Sabin 3 templates. Primer binding sites were spaced along the genome so that the predicted amplification products (142 bp and 163 bp) could be easily resolved electrophoretically from the products generated with our Sabin strain-specific primers (Sabin 1: 97 bp; Sabin 2: 71 bp; Sabin 3: 53 bp). RNAs of all wild type 3 poliovirus isolates from Mexico and Guatemala obtained over a 13-year period (1977-1990) served as efficient templates for amplification of the 142-bp and 163-bp products. Genomic templates derived from vaccine-related polioviruses and most heterologous wild polioviruses were inactive under equivalent reaction conditions. Amplifications generating a 114-bp product with a broadly reacting primer pair, matching highly conserved sequences in the 5'-noncoding region, provided a positive control for the presence in samples of poliovirus (or enterovirus) RNAs. Selective amplification of wild Mexico-Guatemala type 3 poliovirus sequences was obtained with either primer set in reactions containing large stoichiometric excesses (up to 10(6)-fold) of vaccine-related RNAs. We have used wild genotype-specific PCR primer sets to facilitate identification of wild polioviruses present in both clinical and environmental samples.

Amino Acid Sequence↗
PubMed1992

[A case of fatal malabsorption syndrome caused by strongyloidiasis complicated with isosporiasis and human cytomegalovirus infection].

This 54-year-old Korean coal miner suffered from continuous watery diarrhea and weight loss after corticosteroid treatment (beta-methasone, 4 mg daily for 1 week) due to hip-bone fracture in January 1991. Except for the short therapy of steroid, no other histories were contributory. The malabsorption syndrome was aggravated while the case was treated under the impression of amebiasis or intestinal tuberculosis. AIDS antibody test by EIA was negative and quantitative analysis of serum immunoglobulins was in normal ranges. Nine months after the onset of symptoms, the case was diagnosed as malabsorption syndrome caused by complexed and aggravated infection by Strongyloides stercoralis, Isospora and cytomegalovirus in the small intestine, which were proved by stool examination and duodenal biopsy. His clinical course became worse even after high-dosed and prolonged albendazole treatment for strongyloidiasis with supportive fluid therapy. The patient was discharged in hopeless status in November, 1991 and died after one week at home.

Animals↗
PubMed1991

[Preparation and diagnostic application of monoclonal antibodies against Schistosoma japonicum].

The present paper reported on an anti-CCA monoclonal antibody, McAb-IIID 10, which could be used in determinations of both parasite-oriented circulating antigens and specific anti-CCA antibodies. The established competitive ELISA (C-ELISA) using McAb-IIID 10 to detect schistosome-antibodies showed high sensitivity and specificity in the diagnosis of schistosomiasis with few cross-reactions. In field trials, coincident rates in 3 separate batches of serum samples when subjected to double-blind detections were obtained. A total of 1,915 serum samples had been determined by C-ELISA, among them 113 acute cases achieved a 100% positive rate, 765 chronic and 25 late cases showed 96.3% and 72% positive respectively. 70% of the 66 cured schistosomiasis cases turned to be negative. None of the 750 normal individuals showed positive reactions. No cross reaction was found in 27 sera from hydatidosis, whereas 1 and 2 positive reactions were found in 43 paragonimiasis sera and 126 clonorchiasis sera respectively. The established McAb-IIID 10 involved Dot-ELISA was found of value in the assessment of effective chemotherapy and showed a high negative conversion rate of 97.9% in 48 cured schistosomiasis patients. In 16 experimentally infected rabbits, 12 became negative in Dot-ELISA determinations at the 8th week post treatment, and the remaining 4 treated ones, the titer as well as the reaction intensity were also found reduced. A good coincidence rate was also found between C-ELISA and Dot-ELISA, their detection results may be complementary each other.

Animals↗
PubMed1989

Detection of Entamoeba histolytica trophozoites in liver pus by the indirect fluorescent antibody test for the aetiological diagnosis of amoebic liver abscess.

The indirect fluorescent antibody test was used to detect trophozoites of Entamoeba histolytica in liver pus aspirated from patients with amoebic liver abscess. The test can be carried out in no more than two hours. Trophozoites with fluorescence were observed in 17 of the 18 patients with amoebic liver abscesses who were studied. Cells with fluorescence were not found in any negative control specimens from patients with bacterial liver abscess, primary liver cancer, cirrhosis or tuberculous peritonitis. These results indicate that this sensitive, specific and rapid test is very useful in aetiological diagnosis of amoebic liver abscess.

Adult↗
PubMed1989

T lymphocyte subsets in leprosy. A study of 24 Egyptian patients.

Phenotypic analysis was done on 24 Egyptian leprosy patients and 11 healthy controls. The type of leprosy, duration of disease at the time of testing, and age were found to affect T cell subset distribution. As compared with controls, neural leprosy tended to have a decreased total T cell percentage, borderline leprosy an increased T suppressor cell percentage, and reactional borderline leprosy an increased T helper/suppressor ratio. Patients with the disease for less than 1 year had a higher mean percentage of T suppressor cells and a lower mean T helper/suppressor ratio than patients with leprosy for more than 1 year. The same was true in older (50-70 years old) versus younger (12-41 years old) patients.

Adolescent↗