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S T Liu

Publications and source records attributed to S T Liu.

At least 37 records · Page 2Linked to original sources
PubMed1998

Isolation and characterization of plasmid pSW200 from Erwinia stewartii.

The nucleotide sequence of pSW200 of Erwinia stewartii SW2 was determined. This plasmid is 4367 bp long, consisting of four mobilization genes, mobCABD, and an origin of replication homologous to those of ColE1-type plasmids. The plasmid also contains a region of forty-one 15-bp repeats. Deleting this region does not affect the stability or the copy number when maintained as sole plasmid in the cell. However, the plasmid is rapidly lost when a homoplasmid with the intact repeat region is introduced into the cell. The function of this region may provide pSW200 an advantage in competing with an incompatible plasmid in the cell.

Amino Acid Sequence↗
PubMed1998

Transcriptional activation of NF-kappa B activity by Epstein-Barr virus (EBV) LMP1 as a selective therapeutic strategy for EBV-associated diseases.

Epstein-Barr virus (EBV) has been known to be associated with many malignant tumors, including nasopharyngeal carcinoma (NPC). Previous studies have indicated that an EBV-encoded oncoprotein, latent membrane protein 1 (LMP1), is expressed in many NPC tissues. LMP1 has been shown to stimulate HIV LTR through the two NF-kappa B binding sites within this promoter. In this study, we examined the feasibility of using this property of LMP1 as a therapeutic strategy for the treatment of NPC. This therapy consists of the preferential killing of the LMP1-expressing cells by gene transfer using the NF-kappa B-mediated herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) system. The 800-bp HIV-LTR, which contains two NF-kappa B binding sites, was used to drive the HSVtk gene. Stable C33A cell clones expressing the LMP1 and the HSVtk genes were subjected to the GCV sensitivity test. Results showed that cells expressing both the LMP1 and the HSVtk genes were highly sensitive to GCV treatment. These cells were introduced into nude mice subcutaneously and tumors became palpable within 2 weeks. GCV was then introduced intraperitoneally to these mice and the sizes of the tumors were measured daily. Results showed that the tumors regressed in the group of mice carrying cells that stably expressed both the LMP1 and the HSVtk genes, but not in mice carrying cells containing LMP1 or HSVtk alone. Our data indicate that the HSVtk gene expressed from a NF-kappa B-binding motif-containing promoter that is regulated by LMP1 may be used as an in vivo gene therapy strategy of EBV LMP1-expressing cancers such as NPC.

Animals↗
PubMed1998

Characterization of the BcLF1 promoter in Epstein-Barr virus.

The location of the BcLF1 promoter of Epstein-Barr virus (EBV) has been identified by primer extension, which indicates that the +1 site of the BcLF1 mRNA is located at nucleotide 137676 of the EBV genome. According to deletion analysis, the region upstream from nucleotide -38 is not essential for transcription of BcLF1. A 23 bp region in the promoter, from nucleotide -38 to -16, was identified as necessary for regulating the expression of BcLF1, i.e. the promoter activity is activated by 12-O-tetradecanoylphorbol 13-acetate but is repressed by phosphonoacetic acid. The results presented also demonstrate that the oriLyt sequence in cis is essential for enhancing the expression of BcLF1.

Capsid↗
PubMed1998

Molecular cloning and characterization of fengycin synthetase gene fenB from Bacillus subtilis.

A fengycin synthetase gene, fenB, has been cloned and sequenced. The protein (FenB) encoded by this gene has a predicted molecular mass of 143.6 kDa. This protein was overexpressed in Escherichia coli and was purified to near homogeneity by affinity chromatography. Experimental results indicated that the recombinant FenB has a substrate specificity toward isoleucine with an optimum temperature of 25 degrees C, an optimum pH of 4.5, a Km value of 922 microM, and a turnover number of 236 s(-1). FenB also consists of a thioesterase domain, suggesting that this protein may be involved in the activation of the last amino acid of fengycin.

Amino Acid Sequence↗
PubMed1998

Role of Rta in the translation of bicistronic BZLF1 of Epstein-Barr virus.

The BZLF1 gene of Epstein-Barr virus (EBV), which encodes a transcription factor, Zta, is transcribed into monocistronic and bicistronic mRNAs from two different promoters during the immediate-early stage of the EBV lytic cycle. It is generally accepted that the Zta protein translated from the monocistronic mRNA profoundly influences the activation of the EBV lytic cycle. In this study, we constructed a plasmid, pCMV-RZLUC, which can transcribe a bicistronic mRNA consisting of BRLF1 and a BZLF1-luc fusion gene under latent conditions. P3HR1 cells transfected with this plasmid produce a luciferase activity which is approximately 17-fold higher than the activity exhibited by pRZLUC, a plasmid incapable of transcribing the bicistronic mRNA. Genetic analyses indicated that mutations in BRLF1 not only can decrease the translation of the fusion gene from the bicistronic mRNA but can also be complemented by a functional BRLF1 gene in cis. This observation implies that the product of BRLF1, Rta, is involved in the translation of the downstream gene. Results presented herein also demonstrate that these mutations cannot be complemented in trans with a plasmid overexpressing Rta, suggesting that the amount of Rta in the vicinity of the intercistronic region may be crucial for the translation. Furthermore, our results correspond to those of previous investigations indicating that the Zta protein can be translated from the bicistronic mRNA and that, similar to the translation of bicistronic ZLUC, mutations in BRLF1 also hinder the translation of Zta from the BRLF1-BZLF1 bicistronic mRNA. Translation of Zta from the bicistronic mRNA may play an essential role in the activation of the EBV lytic cycle.

Animals↗
PubMed1998

Effects of ciclosporin on whole blood chemiluminescence of renal transplant patients.

AIM: To examine the possible inhibitory role of ciclosporin (Cic) on luminol-dependent chemiluminescence (CL) of whole blood in renal transplant patients. METHODS: Luminol-dependent CL was used to measure active oxygen species generation in respiratory burst of whole blood stimulated by zymosan A. Fluorescence polarization immunoassay was used to monitor the blood concentration of Cic. RESULTS: CL values of Cic group (n = 50) decreased in comparison with those of normal group (n = 10) (P < 0.01). The blood concentration of Cic was negatively related to CL value (P < 0.01). The serum of renal transplant patients directly inhibited respiratory burst of peritoneal macrophages of rats in a concentration-dependent manner. CONCLUSION: Cic inhibits the phagocytic activity of neutrophils in renal transplant patients.

Adult↗
PubMed1997

Cloning and characterization of the ori region of pSW1200 of Erwinia stewartii: similarity with plasmid P1.

The ori region of an Erwinia stewartii plasmid, pSW1200 (106 kb), has been cloned and sequenced. This region consists of a gene encoding a protein which has 91% similarity and 73% identity with the RepA protein of bacteriophage P1. The ori region also consists of eight copies of 19-bp iterons which are highly homologous to the iterons of P1. Similar to plasmid P1, pSW1200 replicon has a copy number of approximately 1. On the other hand, the copy number increases about ninefold if three of the iterons located downstream from repA gene are deleted. We also demonstrate that pGEM-5Z consisting of a copy of P1 iteron is incompatible with a pSW1200 derivative, pSW1201, suggesting that pSW1200 and P1 DNA are incompatible and both belong to the IncY group.

Base Sequence↗
PubMed1997

The presence of RNA splicing signals in the cDNA construct of the E2 gene of classical swine fever virus affected its expression.

E2 is the major neutralizing antigen for classical swine fever virus (CSFV) infection. Previously, we have cloned and sequenced the E2 cDNA of Taiwan strain p97 by the reverse transcription-polymerase chain reaction (RT-PCR) method from CSFV-infected tissue. The presence of RNA splicing donor and acceptor sites were found in the cDNA sequence. In this study, transfection of E2 cDNA into mammalian cells resulted in the production of a spliced RNA. Site-directed mutagenesis of the donor and acceptor sites prevented the RNA splicing event and generated a full length transcript in COS7 cells. Although the spliced E2 transcript has not been reported in natural infection of CSFV, this study suggested that the potential splicing sites affected the E2 gene expression when the plasmid-based E2 gene was introduced into mammalian cells.

Amino Acid Sequence↗
PubMed1997

Measurement of synovial tumor necrosis factor-alpha in diagnosing emergency patients with bacterial arthritis.

Because of the high morbidity and mortality in patients with bacterial arthritis, rapidly and correctly diagnosing this critical condition is a challenge to emergency clinicians. Synovial fluid samples were obtained from 75 patients with arthritis disorders who presented to an emergency service, and levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-6 (IL-6) were measured. Twenty patients with culture-proven bacterial arthritis had higher levels of synovial TNF-alpha than patients with osteoarthritis or with inflammatory arthritis, including gouty arthritis, rheumatoid arthritis, reactive arthritis, and lupus arthritis. There was a good sensitivity for synovial TNF-alpha level in diagnosing patients with bacterial arthritis. Nearly 100% of patients with bacterial arthritis had elevated synovial TNF-alpha levels. However, synovial IL-1 beta and IL-6 levels failed to discriminate bacterial arthritis from other inflammatory arthritis. Measurement of synovial TNF-alpha level may be useful as a diagnostic aid in emergency patients with bacterial arthritis disorders.

Adult↗
PubMed1997

Sequence variations between two Epstein-Barr virus LMP 1 variants have no effect on the activation of NF-kappaB activity.

Previously, we reported that the LMP 1 gene of Epstein-Barr virus (EBV) derived from nasopharyngeal carcinoma (NPC) tissues (i.e., NLMP 1 gene) was able to transform BALB/c3T3 cells. On the other hand, LMP 1 gene of B95-8 strain (i.e., BLMP 1 gene) was not able to transform these cells (Chen et aL, 1992). Further studies indicated that a 10-amino-acid deletion in the carboxyl terminus of NLMP 1 played an important role in transformation (Li et al., 1996). In this study, we tested if this 10-amino-acid deletion affected the induction of NF-kappaB activity by LMP 1. The long terminal repeat of the human immunodeficiency virus type 1 (HIV-1 LTR) contained two copies of NF-kappaB sites and was used to construct the Luc gene-based reporter plasmid, p kappaB-Luc. Plasmid p kappaB-Luc was co-transfected with plasmids containing the NLMP 1 gene, BLMP 1 gene, and their chimeric or deletion constructs, respectively, into C-33A and BALB/c3T3 cells. The activation was then measured by the luciferase activity. Results showed that the full-length proteins induced a similar level of NF-kappaB activity, the two 3' mutants (R15delta and D4delta) still induced a relatively high level of activity, and the two 5' deletion mutants (delta3058 and delta3243) of NLMP 1 gene did not show any significant activation in C-33A cells. However, none of these LMP 1 proteins induced NF-kappaB activity in BALB/c3T3 cells. Using subcellular fractionation analysis and an immunocytostaining method, the truncated proteins of delta3058 and delta3243 were detected in the cytoplasm of the cells whereas the full-length NLMP 1 protein was located at the cytoplasmic membrane. Stable BALB/c3T3 cell clones that expressed both truncated proteins were established and then their ability to induce tumors in nude mice was examined. Data showed that both truncated NLMP 1 proteins still maintained partial transformation activity. Our results suggested that there was no direct correlation between NF-kappaB activation and transformation activity of LMP 1 in BALB/c3T3 cell transformation and that the amino-terminal membrane-spanning domain was important for maintaining both functions of LMP 1.

3T3 Cells↗
PubMed1997

Brachial plexus injury during surgery--report of two cases.

Brachial plexus is the most commonly injured peripheral nerve by malposition during operation. We present two cases of transient brachial palsy after surgery under general anesthesia. Symptoms of the first case persisted about 60 min. Electromyography (EMG) and nerve conduction velocity (NCV) revealed no abnormal finding three days later. In the second case, axonal neuropathy was found at left axillary and suprascapular nerves by EMG and NCV three weeks later. Symptoms persisted for three months and had complete remission after conservative treatment.

Adenocarcinoma↗
PubMed1996

Effect of a 10-amino acid deletion on the oncogenic activity of latent membrane protein 1 of Epstein-Barr virus.

A previous study has shown that the BNLF1 of Epstein-Barr virus (EBV), isolated from a nasopharyngeal carcinoma biopsy (BNLF1-1510), was able to transform Balb/3T3 cells. On the other hand, BNLF1 of a prototype virus B95-8 (BNLF1-958) was not transforming unless the gene was transcribed from a strong promoter. In this study, we have generated chimeric BNLF1 by exchanging the DNA fragments between BNLF1-1510 and BNLF1-958 and examined their expression and transformation ability in Balb/3T3 cells. Results showed that transformation of Balb/3T3 cells by BNLF1-1510 was not due to the excessive expression of the gene. Transfection of Balb/3T3 cells with chimeric BNLF1 showed that the genes with 3' 453 bp sequence of BNLF1-1510 were oncogenic to the cells. Study also revealed that changing the numbers of the 33 bp repeats in the 3' region of the two BNLF1s did not affect the transformation characteristics. On the other hand, deletion of a 30 bp sequence of BNLF1-958, which is absent in BNLF1-1510, changed the gene from non-oncogenic to oncogenic and insertion of this 30 bp sequence into BNLF1-1510 abolished the transformation ability. BNLF1 without this 30 bp sequence was also found in the tumours of other EBV-related neoplastic disease, suggesting that absence of this 30 bp sequence in BNLF1 may be associated with the oncogenesis of these diseases.

3T3 Cells↗
PubMed1996

Characterization of the replicon of plasmid pSW500 of Erwinia stewartii.

A 1.6-kb DNA region required for the replication of pSW500 from Erwinia stewartii SW2 has been identified. DNA sequencing analysis revealed that this DNA fragment consists of a DnaA box, seven 16-bp direct repeats, and a 1005-bp open reading frame. The seven direct repeats have been demonstrated to mediate the incompatibility function of the plasmid. Primer extension analysis showed that the 1005-bp ORF is transcribed in vivo and the +1 site of the transcript is located 113 bp upstream from the translation initiation codon of the ORF. Complementation studies showed that this ORF is required for the replication of the plasmid and may encode a replication protein, RepA. Gene fusion studies revealed that the expression of repA is autoregulated by RepA. We also found that the pSW500 replicon has a copy number of approximately two and that the plasmid is stably maintained in Escherichia coli, thus demonstrating that the replicon contains all the elements required for copy number control and plasmid stability in E. coli. Curing of pSW500 from E. stewartii SW2 revealed that loss of pSW500 did not have any obvious effect on morphology or physiology of the cells, suggesting that pSW500 does not encode a function that is indispensable for the survival of the organism.

Amino Acid Sequence↗
PubMed1996

The study of circulating CD5 positive B lymphocytes in Chinese patients with rheumatoid arthritis.

Circulating CD5+ B lymphocytes were studied in 39 Chinese patients with rheumatoid arthritis (RA). Blood cells were stained with anti-CD5 and anti-CD19 monoclonal antibodies and were analyzed by flow cytometry. The results showed that there was no elevation of CD5+ B cells in RA patients when compared with 41 healthy control subjects. The circulating levels of CD5+ B cell correlated neither with serum titers of rheumatoid factor (RF) nor with disease activities in these patients. The CD5+ B cell levels remained relatively constant after a serial follow-up for 12 months. The similar pattern of epitope density of CD5 antigens also existed in the same patients. Although most studies in Caucasians revealed increased levels of CD5+ B cells in RA patients, measurements of this marker were not useful for the evaluation of disease activities in Chinese patients. Levels of CD5+ B cells may reflect more individual genetic background and may play a minor role in the flare-up of activities in Chinese patients with RA.

Adult↗
PubMed1996

Molecular cloning and nucleotide sequence determination of three envelope genes of classical swine fever virus Taiwan isolate p97.

A strain of classical swine fever virus (CSFV) has been isolated in Taiwan. The cDNA coding for three envelope glycoproteins E1, E2 and E3 were molecularly cloned from purified viral particles using the reverse transcription-polymerase chain reaction (RT-PCR) method and sequence-specific primers. The resulting PCR products (1113 bp for E1. 699 bp for E2 and 567 bp for E3) were cloned into the SmaI site of pUC19 and then subjected to DNA sequence analysis. Data showed that nucleotide sequence of the three envelope genes shared a 82-83% homology with the corresponding genes of three other strains (Alfort, Brescia and Weybridge). However, the homology of the deduced amino acid sequence was greater than 90% among the four strains. The potential asparagine-linked glycosylation sites for E1 (5 sites), E2 (7 sites) and E3 (2 sites) were conserved. This suggests that the Taiwan p97 strain is distinct from other three strains described. The variations may have implications for future vaccine development. The sequence has been submitted to GenBank. The accession numbers are U43924 and U03290.

Amino Acid Sequence↗
PubMed1995

Detection of an Epstein-Barr-virus variant in T-cell-lymphoma tissues identical to the distinct strain observed in nasopharyngeal carcinoma in the Taiwanese population.

An EBV variant has been identified in NPC tissues in Taiwan. This EBV variant contains a point mutation in exon I of the LMP I gene. This mutation results in the loss of an XhoI site at nt 169,426, which is present in strain B95-8. In addition, this variant contains a 30-bp deletion in exon 3 of the gene. The recent demonstration of the prevalence of EBV-containing nasal and peripheral T-cell lymphoma in this region drove us to evaluate the presence of this NPC-EBV strain in 7 cases of T-cell lymphoma, as well as in 48 NPC tissues, 2 cases of Hodgkin's disease and I B-cell lymphoma. Four samples of normal lymph node tissue, 40 of normal nasopharynx tissue and 78 throat washings of healthy individuals were included for comparison. We used sequence-specific primers and the polymerase chain reaction (PCR) method to amplify LMP I gene fragments containing these variations. Mutations were then confirmed by restriction-enzyme digestion and the DNA sequencing analysis. Our results showed that 57 of 58 tumor-tissues samples were EBV-positive. Among them, 56, including 6 T-cell-lymphoma samples, belonged to the NPC strain. This strain of EBV was also present in 92% of EBV-positive normal nasopharynx tissues and in 84% of EBV-positive throat washings of the healthy individuals tested. These results suggest that the NPC-EBV strain is prominently present in Taiwan.

Base Sequence↗
PubMed1995

Functional analysis of EA-D of Epstein-Barr virus.

Different mutations were generated in the diffused-form early antigen (EA-D) of Epstein-Barr virus (EBV) for study of the effects of these mutations in DNA binding, stimulating the activity of EBV-specific DNA polymerase (EBV-DP), and binding to monoclonal antibody R3. Results revealed that the N-terminal 303 amino acids were essential for DNA binding and were sufficient for the enhancement of the activity of EBV-specific DNA polymerase. Deletion study also showed that the region recognized by the R3 monoclonal antibody was located between aa 315 and aa 377. Our results failed to demonstrate the binding between EA-D and EBV-DP, using the proteins synthesized in vitro, suggesting that direct contact between the two proteins is not required for the EBV-DP activity in vitro. We have generated fusion between EA-D and DNA-binding domain of yeast GAL4 protein; however, this fusion protein was not able to transactivate the promoter containing UAS sequence in P3HR1 cells.

Antigens, Viral↗