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Biomedical subjects

J Lin

Publications and source records attributed to J Lin.

At least 559 records · Page 31Linked to original sources
PubMed1995

8-Azido-ATP modification of cytochrome c: retardation of its electron-transfer activity to cytochrome c oxidase.

Horse heart cytochrome c has been modified by 8-azido-ATP and the electron-transfer activity of the modified cytochrome c's to bovine heart cytochrome c oxidase (CcO) under physiological ionic strengths has been studied by the laser flash photolysis technique with 5-deazariboflavin and EDTA as the electron donor. The intermolecular electron transfer between the redox protein partners was shown to be extremely slow. The 8-azido-ATP-modified system exhibited less than 5% of the intracomplex electron-transfer rate observed between native cytochrome c and CcO under otherwise identical conditions. The binding affinity of the modified cytochrome c was greatly reduced (3 orders of magnitude) at low ionic strengths; however, it was only slightly reduced (by a factor of 2) relative to the native protein at physiological ionic strengths. Thus, the binding affinity of the ATP-cytochrome c adducts is relatively insensitive to the ionic strength compared to the native enzyme, suggesting that a different docking conformation is assumed by the ATP-cytochrome c adducts in their interaction with the oxidase. Since the redox potential of the modified cytochrome c is close to the value of its native form, we conclude that there has been a change in the docking of the cytochrome c to CcO and the electronic coupling between heme c and CuA upon 8-azido-ATP modification.

Adenosine Triphosphate↗
PubMed1995

Sarcoma botryoides of the cervix treated with limited surgery and chemotherapy to preserve fertility.

Sarcoma botryoides rarely arises in the uterine cervix. It usually presents in the adolescent age group and is treated primarily by surgery followed by adjuvant chemotherapy. It has been reported in the literature that is possible to limit surgery to local excision in stage I cases, when the tumor is confined to the cervix. In an attempt to preserve reproductive function in a young female patient, we performed cervicectomy alone, followed by adjuvant chemotherapy. After 36 months follow-up she remains well, suggesting that limited excision with adjuvant chemotherapy may be sufficient for early-stage disease.

Adult↗
PubMed1995

Effect of platelet-activating factor on secretion of mucous glycoprotein from chinchilla middle ear epithelial cells in vitro.

Platelet-activating factor (PAF) is a naturally occurring phospholipid that acts as a pleiotropic mediator and mediates cell-cell reactions under physiological and pathological conditions. Recently, it has been shown that PAF is a strong secretagogue of mucous glycoprotein in the airways, suggesting its role in mucous glycoprotein secretion and the pathogenesis of otitis media with effusion. In the current study, we examined the effect of PAF on mucous glycoprotein secretion in cultured chinchilla middle ear epithelial cells. PAF at 1 microM significantly stimulated mucous glycoprotein secretion from cultured chinchilla middle ear epithelial cells. This action was concentration-dependent, with secretions reaching near maximum when the cells were incubated with PAF at 100 microM. In a time-dependent study, PAF demonstrated an initial rapid stimulation of mucous glycoprotein secretion, followed by a gradual increase thereafter. A six-fold increase was seen in the first 2 h compared with controls. Cycloheximide, a protein synthesis inhibitor, demonstrated an inhibitory effect on PAF-stimulated mucous glycoprotein secretion in this study. These findings suggest that PAF plays an important role in the pathogenesis of otitis media with effusion by stimulating mucous glycoprotein secretion in vitro.

Animals↗
PubMed1995

Quantitative evaluation of thallium-201 uptake in predicting chemotherapeutic response of osteosarcoma.

Thallium-201 has been shown to be useful in predicting tumour viability in patients undergoing neoadjuvant chemotherapy for osteogenic sarcoma. Early studies relied upon qualitative assessment of analog images to obtain predictive results. Recently, the lesion to normal tissue uptake ratio of 201Tl has been used in evaluating bone and soft tissue sarcomas. This study attempts to quantitate changes in tumour to normal tissue ratio following chemotherapy. Eight consecutive patients with classical osteosarcoma received standard preoperative chemotherapy with a combination of cisplatin, adriamycin and high-dose methotrexate. 201Tl gamma scintigraphic images were obtained both before and after chemotherapy. The average counts taken over the tumour divided by that from the contralateral normal tissue area yielded a tumour-to-normal tissue (T/N) ratio. The percentage change in the T/N ratio before and after preoperative chemotherapy was correlated with the percentage of tumour necrosis from pathological section. The median post-chemotherapy T/N ratio was 1.85 (range 0.5-7.7). The median percentage change in T/N ratio after chemotherapy was -58% (range +26% to -83%). The median percentage of necrosis from pathological section was 80% (range 0%-95%). There was a good correlation between the percentage of tumour necrosis and the percentage change in T/N ratio (rank correlation coefficient r = 0.84, P = 0.0085). Quantitative assessment of changes in 201Tl uptake by osteosarcoma correlates well with tumour necrosis after preoperative chemotherapy. This method may be used to predict response to chemotherapy at an earlier stage, enabling the clinician to consider alternative chemotherapeutic regimens or salvage surgery.

Adolescent↗
PubMed1995

Interruption of a transforming growth factor alpha autocrine loop in Caco-2 cells by antisense oligodeoxynucleotides.

BACKGROUND & AIMS: We have previously shown that Caco-2 cell proliferation is driven by basolateral membrane epidermal growth factor receptors. The aim of this study was to investigate whether autocrine production of transforming growth factor alpha (TGF-alpha) activates these receptors and stimulates proliferation using antisense oligodeoxynucleotides. METHODS: Caco-2 cells grown on microporous membranes or Jurkat cells were exposed to conventional or 5' cholesterol-modified oligodeoxynucleotides synthesized with random, antisense, or missense base sequences. Indices of proliferation were measured, including [3H]thymidine or [3H]uridine uptake for studies of short-term stimulation and the methylthiotetrazole assay as an index of cell number increase over longer periods. Secretion of TGF-alpha by cells was detected using a soft agar bioassay. RESULTS: Incubation with antisense oligodeoxynucleotides inhibited TGF-alpha secretion compared with controls. Random and missense oligodeoxynucleotides had no effect on proliferation. The TGF-alpha antisense oligodeoxynucleotides markedly inhibited proliferation, an effect that was abolished by adding TGF-alpha to the medium. Oligonucleotides had no effect on Jurkat cells, a lymphocytic cell line lacking epidermal growth factor receptors. Cholesterol-modified oligodeoxynucleotides were more effective and specific than unmodified oligodeoxynucleotides. CONCLUSIONS: Caco-2 cell proliferation is driven by autocrine stimulation of epidermal growth factor receptors by TGF-alpha. This mechanism may be effectively inhibited by antisense oligodeoxynucleotides, particularly those modified by the 5' attachment of cholesterol.

Base Sequence↗
PubMed1995

Vaccination of sheep against Schistosoma japonicum with either glutathione S-transferase, keyhole limpet haemocyanin or the freeze/thaw schistosomula/BCG vaccine.

The protective potential of glutathione S-transferase (GST), keyhole limpet haemocyanin (KLH) and the freeze/thaw (F/T) schistosomula/BCG vaccine was evaluated against Schistosoma japonicum in the natural sheep host. Groups of ten sheep each were vaccinated as follows: Group I: 2 x F/T 30,000 schistosomula+BCG 3 x 10(8) organisms, with a 2 week interval between vaccinations (F/T 'Low'). Group II: 3 x F/T 20,000 schistosomula+BCG 3 x 10(8), with 4 week interval (F/T 'High'). Group III: 2 x GST 0.24 mg+FCA (Freund's complete adjuvant) with 2 week interval (GST 'Low'). Group IV: 3 x GST 0.24 mg+FCA, with 4 week interval (GST 'High'). Group V: 2 x KLH 1.0 mg in phosphate-buffered saline (PBS), with 2 week interval (KLH 'Low'). Group VI: 3 x KLH 1.0 mg in PBS, with 4 week interval (KLH 'High'). Group VII: control (not vaccinated). Specific antibody, detected by GST-enzyme-linked immunosorbent assay (ELISA) and KLH-ELISA on the day after the last vaccination and 1, 2 and 3 weeks post-challenge, was found in all GST- or KLH-vaccinated groups. The same was found in F/T schistosomula-vaccinated groups against crude adult worm antigen (AWA). In Western blotting all GST-vaccinated sera recognized 26 kDa and 28 kDa bands on the challenge day and at 3 and 11 weeks post-challenge. Mean faecal egg counts between Weeks 6 and 10 post-challenge were reduced in a statistically significant way at five time points in the four groups, i.e. 83.38% (P < 0.005) in Group II, 49.29% (P < 0.025) in Group III, 47.9% (P < 0.05) and 71.15% (P < 0.01) in Group IV, 52.0% (P < 0.025) and 66.38% (P < 0.025) in Group VI. On autopsy and perfusion 1 week after the last faecal count, adult worm reductions were obtained of 40.36% (P < 0.05) in Group I, 37.26% (P < 0.025) in Group II, 24.73% (not significant) in Group III, 35.93% (P < 0.025) in Group IV, 27.46% (P < 0.05) in Group V and 33.81% (P < 0.01) in Group VI. Mean tissue egg densities were also reduced significantly in Groups III, IV and VI, especially in Group IV vaccinated animals. Mean liver egg granuloma diameters of the vaccinated groups were found to be less than those of the controls but there was no statistical significance.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Percutaneous trans-hepatic cholangioscopy and lithotripsy in the treatment of intrahepatic stones: a study with 5 year follow-up.

BACKGROUND: Intrahepatic stones are characterized by high treatment failure and recurrence rates. In the past, surgery played a major role in the management of this disease, but surgical intervention may not be feasible in previously operated patients or those classified as poor surgical risks. The development of percutaneous trans-hepatic cholangioscopy and lithotripsy has played an important role in the treatment of these patients. METHODS: We reviewed our results in 165 patients with intrahepatic stones treated by percutaneous trans-hepatic cholangioscopy and lithotripsy in the past 12 years. RESULTS: The rate of complete stone removal was 80% (132 cases) and the rate of recurrent biliary stones after a mean 58 month follow-up interval was 32.6% (43 of 132 cases). There were 2 deaths during the treatment period and 10 deaths during follow-up. CONCLUSIONS: Percutaneous trans-hepatic cholangioscopy and lithotripsy are good alternatives in treating primary intrahepatic stones, especially for those patients unsuitable for surgery.

Bile Ducts, Intrahepatic↗
PubMed1995

Analysis of a conformation-independent epitope and a conformational epitope in a protein: a study on cobrotoxin from Taiwan cobra venom.

The antibodies against cobrotoxin were separated into two antibody preparations by successive affinity chromatographies on reduced and S-carboxymethylated (RCM)-cobrotoxin-Sepharose, and cobrotoxin-Sepharose columns. The antibodies (abbreviated as Abcf-i) that bound with the RCM-cobrotoxin-Sepharose were verified to specifically recognize the continuous epitopes of cobrotoxin, which were insensitive to conformational changes. Whilst the antibodies (abbreviated as Abcf-d) that did not bind with the RCM-cobrotoxin-Sepharose column recognized the conformational epitopes in cobrotoxin. The two antibody preparations were employed to screen the antigenic peptides derived from the proteolytic hydrolysate of cobrotoxin and RCM-cobrotoxin. Five antigenic peptides (AP-4, AP-5, AP-10, AP-11, and AP-12) were obtained from the acid protease A-digested hydrolysate of cobrotoxin, and two antigenic peptides (V8-2 and V8-4) were found in the hydrolysate of RCM-cobrotoxin after hydrolysis with Saccharomyces aureus V8 protease. The segments at positions 1-21 and 22-38 encompassed the peptide fractions, AP-4, AP-5, V8-2, and V8-4, that reacted with Abcf-i, indicating that the two segments bore the continuous epitopes of cobrotoxin. Alternatively, AP-10, AP-11, and AP-12 reacted with both Abcf-i and Abcf-d. The structures of the three peptides had a common segment at positions 43-62, suggesting that this region comprised the conformation-independent epitopes as well as conformational epitopes in cobrotoxin. These results reflected that the conformation-independent and conformational epitopes in a protein can be separately identified.

Amino Acid Sequence↗
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

The stationary-phase sigma factor sigma S (RpoS) is required for a sustained acid tolerance response in virulent Salmonella typhimurium.

The acid tolerance response (ATR) of log-phase Salmonella typhimurium is induced by acid exposures below pH 4.5 and will protect cells against more extreme acid. Two systems are evident: a transiently induced system dependent on the iron regulator Fur that provides a moderate degree of acid tolerance and a more effective sustained ATR that requires the alternate sigma factor sigma S encoded by rpoS. Differences between the acid responses of virulent S. typhimurium and the attenuated laboratory strain LT2 were attributed to disparate levels of RpoS caused by different translational starts. The sustained ATR includes seven newly identified acid shock proteins (ASPs) that are dependent upon sigma S for their synthesis. It is predicted that one or more of these ASPs is essential for the sustained system. The sustained ATR also provided cross-protection to a variety of other environmental stresses (heat, H2O2 and osmolarity); however, adaptation to the other stresses did not provide significant acid tolerance. Therefore, in addition to starvation, acid shock serves as an important signal for inducing general stress resistance. Consistent with this model, sigma S proved to be induced by acid shock. Our results also revealed a connection between the transient and sustained ATR systems. Mutations in the regulator atbR are known to cause the overproduction of ten proteins, of which one or more can suppress the acid tolerance defect of an rpoS mutant. One member of the AtbR regulon, designated atrB, was found to be co-regulated by sigma S and AtbR. Both regulators had a negative effect on atrB expression. The results suggest AtrB serves as a link between the sustained and transient ATR systems. When sigma S concentrations are low, a compensatory increase in AtrB is required to engage the transiently induced, RpoS-independent system of acid tolerance. Results also suggest different acid-sensitive targets occur in log-phase versus stationary-phase cells.

Acids↗
PubMed1995

Video-assisted thoracoscopic wedge resections of pulmonary metastatic osteosarcoma: should it be performed?

We studied the use of video-assisted thoracoscopic (VAT) surgery in the management of metastatic osteosarcoma. From September 1993 to March 1994, we performed a total of 11 VAT wedge resections of pulmonary metastatic osteosarcoma in seven patients (six males, one female, age 12 to 46 years). Three patients had bilateral procedures performed either under the same anaesthesia or in stages. One patient had two operations on the same side. The average number of nodules excised was three. Two patients subsequently required formal lobectomies when the metastatic tumours were either too big or too close to the hilum for safe wedge resections. There was one death on postoperative day 3 due to dysrhythmia. One patient died 5 months later from a progression of his underlying disease. Two patients remained disease free up to 8 and 12 months, respectively, from their first operations. The average postoperative chest drain duration was 1.4 +/- 0.7 days and hospital stay 2.3 +/- 1.1 days. The procedure was well tolerated and postoperative morbidity was minimal. We conclude that although VAT wedge resection of pulmonary metastatic osteosarcoma is feasible technically and is associated with a short hospital stay and minimal morbidity, this approach cannot be recommended when complete resection of all metastases is the goal as the technique relies heavily on computed tomographic scans to detect nodules. Recurrence of metastasis from 4 to 6 months in three of seven patients argues against VAT surgery being an adequate procedure. The high cost of the staplers, in addition, is a secondary consideration.

Adolescent↗
PubMed1995

Protein synthesis in Brucella abortus induced during macrophage infection.

Brucella abortus is a facultative, intracellular, pathogenic bacterium that replicates within macrophages and resists macrophage microbicidal mechanisms. To study gene expression and to elucidate the defense mechanisms used by B. abortus to resist destruction within macrophages, protein synthesis by B. abortus was examined by pulse-labeling techniques during intracellular growth within J774A.1, a macrophage-like cell line. Prominent changes observed include increased synthesis of Brucella proteins with estimated molecular masses of 62, 28, 24, and 17 kDa. The 62-kDa protein was identified by immunoprecipitation analysis as Hsp62, a GroEL homolog. A protein of 60 kDa was expressed during acid shock and may represent a modified form of Hsp62. The 28- and 17-kDa proteins have not been observed under any in vitro stress condition and presumably represent macrophage-specific induction. The 24-kDa protein comigrates with an unidentified protein induced by acid shock, designated Asp24. Expression of Asp24 is optimal at pH values below 4.0 and within the first 3 h following a shift from pH 7.3 to 3.8. This corresponds directly with a period of optimal bacterial survival at a reduced pH and suggests an active role for this protein in resistance to such environments. The identification of these gene products and the mechanisms controlling their expression is an important step in understanding the resistance of Brucella spp. to intracellular destruction within macrophages.

Animals↗
PubMed1995

Comparative analysis of extreme acid survival in Salmonella typhimurium, Shigella flexneri, and Escherichia coli.

Several members of the family Enterobacteriaceae were examined for differences in extreme acid survival strategies. A surprising degree of variety was found between three related genera. The minimum growth pH of Salmonella typhimurium was shown to be significantly lower (pH 4.0) than that of either Escherichia coli (pH 4.4) or Shigella flexneri (pH 4.8), yet E. coli and S. flexneri both survive exposure to lower pH levels (2 to 2.5) than S. typhimurium (pH 3.0) in complex medium. S. typhimurium and E. coli but not S. flexneri expressed low-pH-inducible log-phase and stationary-phase acid tolerance response (ATR) systems that function in minimal or complex medium to protect cells to pH 3.0. All of the organisms also expressed a pH-independent general stress resistance system that contributed to acid survival during stationary phase. E. coli and S. flexneri possessed several acid survival systems (termed acid resistance [AR]) that were not demonstrable in S. typhimurium. These additional AR systems protected cells to pH 2.5 and below but required supplementation of minimal medium for either induction or function. One acid-inducible AR system required oxidative growth in complex medium for expression but successfully protected cells to pH 2.5 in unsupplemented minimal medium, while two other AR systems important for fermentatively grown cells required the addition of either glutamate or arginine during pH 2.5 acid challenge. The arginine AR system was only observed in E. coli and required stationary-phase induction in acidified complex medium. The product of the adi locus, arginine decarboxylase, was responsible for arginine-based acid survival.

Adaptation, Physiological↗