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Biomedical subjects

S F Lin

Publications and source records attributed to S F Lin.

At least 37 records · Page 2Linked to original sources

Autologous peripheral blood stem cells harvesting in Kaohsiung Medical University Hospital.

We report our clinical experience in autologous peripheral blood stem cells harvesting. A total of 40 patients with 112 apheresis procedures were analyzed, 88 with Cobe system and 24 with MCS3P system. Our results revealed that there was no significant difference in efficiency of CD34(+) cell harvesting between both apheresis systems, but the Cobe system had more nucleated cells collected and less red cell contaminated in the final PBSCs collections. The percentage of CD34(+) cells collected decreased significantly following the first day's harvesting (p = 0.026). There was a good correlation between the percentage of CD34(+) cells in PBSCs and colony forming units-granulocyte macrophage (CFU-GM) or burst forming unit-erythrocyte (BFU-E)(r = 0.909, p < 0.0001; r = 0.788, p < 0.0001, respectively). However, it was negatively correlated with the patient's age. The CD34(+) cells collected in patients with acute leukemias were also higher than those patients with other solid tumors. Ten cases (13%) with 15 apheresis procedures experienced side effects like numbness, nausea, fever, or headache etc. The Cobe system seemed to have higher frequency of side effects than that of MCS3P system (16% vs 4%). From our results, we concluded that both COBE and MCS3P system have similar efficiency and all patients could tolerate the apheresis procedures in peripheral blood stem cells harvesting. The CD34(+) cell can be used as a good parameter to estimate the amount of stem cells collected. The patient's age and disease pattern were significant factors influencing the CD34(+) stem cells collection in autologous peripheral blood stem cells harvesting.

Adolescent↗

[Isolation of a kind of endophytic fungus which can produce taxol compounds].

OBJECTIVE: To isolate a kind of endophytic fungus which can produce taxol compounds from the bark of Taxus Cuspidata Sieb et Zucc. METHODS: Endophytic fungus was isolated from the bark of Taxus Cuspidata Sieb et Zucc by aseptic techniques and the analysis of the culture fluid from the fermentation of fungus was carried out by high performance liquid chromatography (HPLC). RESULTS: Endophytic fungus was obtained from the bark of Taxus Cuspidata Sieb et Zucc and the chemical analysis showed that the culture of fungus contain taxol compounds. CONCLUSION: Endophytic fungus which can produce taxol compounds exists in the bark of Taxus Cuspidata Sieb et Zucc.

Chromatography, High Pressure Liquid↗

The protein tyrosine kinase family of the human genome.

As the sequencing of the human genome is completed by the Human Genome Project, the analysis of this rich source of information will illuminate many areas in medicine and biology. The protein tyrosine kinases are a large multigene family with particular relevance to many human diseases, including cancer. A search of the human genome for tyrosine kinase coding elements identified several novel genes and enabled the creation of a nonredundant catalog of tyrosine kinase genes. Ninety unique kinase genes can be identified in the human genome, along with five pseudogenes. Of the 90 tyrosine kinases, 58 are receptor type, distributed into 20 subfamilies. The 32 nonreceptor tyrosine kinases can be placed in 10 subfamilies. Additionally, mouse orthologs can be identified for nearly all the human tyrosine kinases. The completion of the human tyrosine kinase family tree provides a framework for further advances in biomedical science.

Animals↗

Mechanisms of ventricular fibrillation induction by 60-Hz alternating current in isolated swine right ventricle.

BACKGROUND: The mechanisms by which 60-Hz alternating current (AC) can induce ventricular fibrillation (VF) are unknown. METHODS AND RESULTS: We studied 7 isolated perfused swine right ventricles in vitro. The action potential duration restitution curve was determined. Optical mapping techniques were used to determine the patterns of activation on the epicardium during 5-second 60-Hz AC stimulation (10 to 999 microA). AC captured the right ventricles at 100+/-65 microA, which is significantly lower than the direct current pacing threshold (0.77+/-0.45 mA, P:<0.05). AC induced ventricular tachycardia or VF at 477+/-266 microA, when the stimulated responses to AC had (1) short activation CLs (128+/-14 ms), (2) short diastolic intervals (16+/-9 ms), and (3) short diastolic intervals associated with a steep action potential duration restitution curve. Optical mapping studies showed that during rapid ventricular stimulation by AC, a wave front might encounter the refractory tail of an earlier wave front, resulting in the formation of a wave break and VF. Computer simulations reproduced these results. CONCLUSIONS: AC at strengths less than the regular pacing threshold can capture the ventricle at fast rates. Accidental AC leak to the ventricles could precipitate VF and sudden death if AC results in a fast ventricular rate coupled with a steep restitution curve and a nonuniform recovery of excitability of the myocardium.

Animals↗

Dihydrofolate reductase from Kaposi's sarcoma-associated herpesvirus.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the first human virus known to encode dihydrofolate reductase (DHFR), an enzyme required for nucleotide and methionine biosynthesis. We have studied the purified KSHV-DHFR enzyme in vitro and analyzed its expression in cultured B-cell lines derived from primary effusion lymphoma (PEL), an AIDS-associated malignancy. The amino acid sequence of KSHV-DHFR is most similar to human DHFR (hDHFR), but the viral enzyme contains an additional 23 amino acids at the carboxyl-terminus. The viral DHFR, overexpressed and purified from E. coli, was catalytically active in vitro. The K(m) of KSHV-DHFR for dihydrofolate (FH(2)) was 2.4 microM, which is significantly higher than the K(m) of recombinant hDHFR (rhDHFR) for FH(2) (390 nM). K(m) values for NADPH were similar for the two enzymes, about 1 microM. KSHV-DHFR was inhibited by folate antagonists such as methotrexate (K(i): 200 pM), aminopterin (K(i): 610 pM), pyrimethamine (K(i): 29 nM), trimethoprim (K(i): 2.3 microM), and piritrexim (K(i): 3.9 nM). In all cases, K(i) values for these folate antagonists were higher for KSHV-DHFR than for rhDHFR. The viral enzyme was expressed at levels two- to tenfold higher than hDHFR in PEL cell lines as an early lytic cycle gene. KSHV-DHFR mRNA and protein appeared from 6 to 24 h after chemical induction of the KSHV lytic cycle. Epitope-tagged KSHV-DHFR and rhDHFR both localized to the nucleus of transfected cells, while other KSHV nucleotide metabolism genes localized to the cytoplasm. DHFR activity was not essential for viral replication in cultured PEL cells. Since hDHFR was not detectable in peripheral blood mononuclear cells (PBMCs), KSHV-DHFR may function to provide increased DHFR activity in vivo in infected cells that have little or none of their own enzyme.

Amino Acid Sequence↗

Mutation analysis of PTEN/MMAC1 in acute myeloid leukemia.

Recently, a putative tumor suppressor gene, PTEN/MMAC1, has been identified at chromosome 10q23.3, which encodes a 403 amino acid dual-specificity phosphatase containing a region of homology to tensin and auxillin. Somatic mutations of the PTEN/MMAC1 gene have been identified in a number of cancer cell lines and primary cancers. Mutations in PTEN/MMAC1 are most frequently found in advanced cancers. To evaluate the role of the PTEN/MMAC1 gene in leukemia, bone marrow and/or peripheral blood from 62 acute myeloid leukemia (AML) patients, 5 hemopoietic cell lines (HL60, U937, Raji, KG-1, K562), and 30 normal controls were analyzed. The results showed aberrant PTEN/MMAC1 transcripts in 15 of the 62 (24%) AML patients, 4 of the 5 cell lines (80%), and 4 of the 30 (13%) normal controls. As in our previous study of TSG101, the abnormal transcripts may result from aberrant RNA splicing as evidenced by the presence of both these aberrant transcripts and normal full length transcripts in all specimens examined. Loss of heterozygosity (LOH) analysis and PCR-SSCP of the entire coding region showed that none of the AML cases had LOH or mutation. Only one frameshift mutation at codon 130 (insertion of CCCG) with premature termination of coding sequence was observed in the U937 cell line. Our results indicate that the PTEN/MMAC1 gene may play a role in a small percentage of AML, but its significance needs to be further evaluated.

Acute Disease↗

Lewis (FUT3) genotypes in Taiwanese, Thai, and Filipino populations.

The Lewis (Le) blood type comprises two major antigens, Le(a) and Le(b), which are encoded by alpha (1,2)-fucosyltransferase (FUT2) and a (1,3/1,4)-fucosyl-transferase (FUT3). In this study, we analyzed the mutations of FUT3 in Taiwanese, Thai, and Filipino populations and correlated these with serologic phenotypes. One hundred and thirty-seven Taiwanese, 71 Thai, and 125 Filipino were studied unselectively. The frequency of the normal and four other mutant alleles for Taiwanese, Thai, and Filipino, respectively, were as follows: 187/274 (68.2%), 87/142 (61.3%), and 160/250 (64.0%) were wild type (Le); 14/274 (5.1%), 1/142 (0.7%), and 1/250 (0.4%) were a T202C/C314T mutation (le202,314); 35/274 (12.8%), 15/142 (10.6%), and 22/250 (8.8%) had the G508A mutation (le508); and 38/274 (13.9%), 39/142 (27.4%), and 67/250 (26.8%) carried the T1067A mutation (le1067). The le445 and le1007 were not detected in this study. Our result provided the first genetic data of the FUT3 gene in these three populations, and the frequency distribution of mutant alleles among Taiwanese, Thai, and Filipinos demonstrates a significant difference (P<0.001). In our study, the le202,314 mutation had considerable frequency in the Taiwanese, but the le1067 mutation had a higher frequency in Thai and Filipinos.

Adult↗

Three-dimensional surface reconstruction and fluorescent visualization of cardiac activation.

Optical imaging of transmembrane potentials in cardiac tissue is a rapidly growing technique in cardiac electrophysiology. Traditional studies typically use a monocular imaging setup, thus limiting investigation to a restricted region of tissue. However, studies of large-scale wavefront dynamics, especially those during fibrillation and defibrillation, would benefit from visualization of the entire epicardial surface. To solve this problem, a panoramic cardiac visualization algorithm was developed which performs the two tasks of reconstruction of the surface geometry of the heart, and representation of the panoramic fluorescence information as a texture mapping onto the geometry that was previously created. This system permits measurement of epicardial electrodynamics over a geometrically realistic representation of the actual heart being studied. To verify the accuracy of the algorithm, the procedure was applied to synthetic images of a patterned ball; further verification was provided by application of the algorithm to a model heart placed in the experimental setup. Both sets of images produced mean registration image errors on the order of 2 pixels, corresponding to roughly 3 mm on the geometry. We demonstrate the algorithm by visualizing epicardial wavefronts on an isolated, perfused rabbit heart.

Algorithms↗

Clinical implications of aberrant TSG101 transcripts in acute myeloblastic leukemia.

Tsg101 is a mouse tumor suppressor gene whose homozygous deletion produces transformation of NIH3T3 cells and leads to metastases in nude mice. The human homologue of the gene, TSG101, is localized in chromosome 11p15.1-p15.2. Reduced TSG101 expression may cause the defect of the cell cycle checkpoint that leads to genetic instability and consequently to the progression of neoplasia. Aberrant TSG101 transcript have been identified in many types of cancers, and the relaxation of RNA splicing fidelity may be an onco-developmental marker in cancers and could play a general role in tumorigenesis. In our previous study, smaller TSG101 transcripts were found in AML specimens, hematopoietic cell lines and normal controls. The aberrant transcripts occurred more frequently in the AML cases and cell lines. The patients with aberrant TSG101 transcripts had higher initial white cell count, lower LDH level, and lower complete remission rate after induction chemotherapy. However, further multivariate analysis of clinical data revealed that there was no relationship to the TSG101 aberrant transcripts. The clinical significance of TSG101 aberrant transcript in AML needs further evaluation.

3T3 Cells↗

Aberrant transcripts of FHIT, TSG101 and PTEN/MMAC1 genes in normal peripheral mononuclear cells.

Aberrant transcripts of FHIT and TSG101 using nested RT-PCR were reported in many human tumours. The role of these aberrant transcripts in tumourigenesis is not clear. We, therefore, analyzed the aberrant transcripts of FHIT, TSG101 and PTEN/MMAC1 in peripheral mononuclear cells of normal individuals using nested RT-PCR to explore the role of these genes in cancer development. The results showed that there are at least five types of aberrant transcripts: type I is the deletion at junction located in-between normal exon and intron; type II has deletion of some bases and subsequent insertion of several bases in the deletion area; type III accommodates splicing donor or acceptor site-like sequence; type IV has homologous sequences near the deleted junction; and type V comprises the homologous sequences at the deletion junction. A normal healthy person can have more than one aberrant transcripts of FHIT, TSG101 and PTEN/MMAC1 genes. The size and the number of the transcripts vary and the diversity is unconstrained. It is not depended on the time, condition of the reaction, or the isolation method. From these results, we suggested that the aberrant transcripts of FHIT, TSG101 and PTEN/MMAC1 genes may be the imperfect products of splicesome which occur one in every thousands, ten thousands or more. As a result, these data implied no direct association between the aberrant transcripts and tumourigenesis.

Acid Anhydride Hydrolases↗

Mutation analysis of PTEN/MMAC 1 in sporadic thyroid tumors.

Recently, a putative tumor suppressor gene, PTEN/MMAC1, has been identified at chromosome 10q23.3. This gene encodes a 403 amino acid dual specificity phosphatase containing a region of homology to tensin and auxillin. Somatic mutations of the PTEN/MMAC1 gene have been found in a number of cancer cell lines and primary cancers. Cowden disease, an autosomal dominant harmartoma syndrome associated with thyroid and breast tumors, has been found to be associated with mutations of PTEN/MMAC1 gene. To evaluate the role of the PTEN/MMAC1 gene in sporadic thyroid tumors, we studied 17 sporadic thyroid tumors, of which 12 were papillary thyroid carcinomas, 1 was follicular thyroid carcinoma, 1 was medullary thyroid carcinoma and 3 were thyroid adenomas. Direct sequencing of PCR-amplified products was performed for all nine exons of PTEN/MMAC1. No mutations of PTEN/MMAC1 gene were observed in any of the sporadic thyroid tumors. Our results indicate that the PTEN/MMAC1 gene may not play a major role in sporadic thyroid tumors.

Genes, Tumor Suppressor↗

Molecular basis of Lewis blood type in Taiwanese.

The Lewis (Le) histo-blood group system comprises two major antigens, Le(a) and Le(b) which are determined by alpha (1,2)-fucosyltransferase (FUT2) and alpha (1,3/1,4)-fucosyltransferase (FUT3). In this study, we analyzed the mutations of FUT2 and FUT3 genes in 101 Taiwanese by molecular biology method and compared them with their serologic phenotypes to explore their relationship. There is at least one wild allele of FUT2 and FUT3 genes in phenotype of Le (a-b+). The phenotypes of Le (a+b-) and Le (a+b+) are caused by mutations of both alleles of FUT2 gene and at least one wild allele of FUT3 gene. The genotypes of Le (a+b-) and Le (a+b+) are the same. Twenty cases are phenotype of Le (a-b-), which are caused by mutations of both alleles of FUT 2 gene and/or FUT 3 gene. Twelve cases were caused by both alleles mutations of FUT 3 gene only, while three cases were caused by mutations of both alleles of FUT2 gene and the rest of the cases were caused by mutations of both alleles of FUT2 and FUT3 genes. Our findings confirm that the Le histo-blood group is determined by the interaction of FUT 2 and FUT 3 genes. Our report is the first study of FUT 2 gene and FUT 3 gene in a Taiwanese population. We suggest that the genetic analysis of Le blood group should include FUT 2 and FUT 3 genes together.

Fucosyltransferases↗

[Analysis of imidacloprid and damanlin by high performance liquid chromatography].

A new method for determination of imidacloprid and damanlin by using HPLC was established. It was carried out on a mu Bondapak C18 column (3.9 mm i.d. x 300 mm) with ACN-MeOH-H2O (60:10:30, V/V) as eluent and detected at 240 nm using photodiode array detector. The results of imidacloprid and damanlin were 3.71% and 15.65% respectively. The standard deviations for imidacloprid and damanlin were 0.88% and 0.76% respectively.

Animals↗

Molecular characterization of secretor type alpha(1, 2)-fucosyltransferase gene deficiency in the Philippine population.

We analyzed the seven mutations which are responsible for the deficiency of the secretor type alpha(1,2)-fucosyltransferase gene product, Se enzyme, in the Philippine population. One hundred and one unrelated Filipinos in Taiwan were studied. A new mutation, a 3-base pair deletion from nt 688 through 690, was found in two (0. 1%) of 202 chromosomes. The frequencies of six other mutated alleles were as follows: 71/202 (35.2%) were cDNA 385 A-->T missensed mutation (se2), 28/202 (13.9%) were C571T nonsense mutation (se3), 16/202 (7.9%) were G849A nonsense mutation (se4), 4/202 (1.9%) were G428A nonsense mutation (se1), and 81/202 (40.1%) were wild-type allele (Se). No C628T nonsense mutations (se5) or fusion genes of pseudogene and FUT2 gene (se 6) were found in this population. For the molecular basis of phenotype Le(a+ b-): eight cases had se2/se2, six cases had se2/se3, two cases had se3/se4, one case was homozygous of se4, one case was se3/se1, and two cases were se2/se7. For the Le(a+ b+) phenotype: four cases had se2/se2, two cases had se2/se3, one case was se3/se3, and one case was se2/se4. For the Le(a- b+) phenotype: 16 cases were Se/Se, 21 cases were Se/se2, six cases were Se/se3, five cases were Se/se4, and two cases had Se/se1. Our results suggest that the genotypes of the alpha(1, 2)-fucosyltransferase gene in phenotypes Le(a+ b+) and Le(a+ b-) are the same. Other factors that play important roles may cause the differences between these two phenotypes. Several hotspot mutations in the alpha(1,2)-fucosyltransferase gene are responsible for the nonsecretor phenotype.

Fucosyltransferases↗

Excavatolides F-M, new briarane diterpenes from the gorgonian Briareum excavatum.

Eight new briarane-type diterpenes, excavatolides F-M (1-8), have been isolated from the gorgonian Briareum excavatum. The structures and relative stereochemistry of these compounds were established by spectral analysis and chemical methods. The cytotoxicity of these compounds toward various cancer cell lines has also been determined.

Animals↗

Quatrefoil reentry in myocardium: an optical imaging study of the induction mechanism.

INTRODUCTION: The "critical point hypothesis" for induction of ventricular fibrillation has previously been extended to infer the coexistence of four critical points, and hence four simultaneous spiral reentries or a quatrefoil reentry, resulting from only one premature stimulus delivered to the same location as the pacing stimulus. An optical imaging technique was used to explore its existence and to study the induction mechanism of this peculiar reentry pattern. METHODS AND RESULTS: In 16 isolated, Langendorff-perfused rabbit hearts, high-speed optical imaging at 133 or 267 frames/sec was performed to observe the induced response with a unipolar point electrode. A novel quatrefoil-shaped reentry pattern consisting of two pairs of opposing rotors was created by delivering long stimuli during the vulnerable phase. Successful induction occurred in a narrow range of coupling intervals. A dogbone pattern of virtual electrodes was established during the premature stimulus. Propagating wavefronts launched from the virtual anodes immediately after the termination of S2. The alternating blocking and conducting effects of the virtual electrodes, as well as the boundary between virtual cathode and virtual anode, provided the necessary pathways for quatrefoil reentry. Propagation directions of the reentrant spiral wavefronts reversed with a reversal in S2 polarity. Quatrefoil reentries were not sustained and lasted 1 to 4 complete cycles. CONCLUSION: The initiation of quatrefoil reentry followed anodal- or cathodal-break stimulation as a result of local symmetrical enhancement of the dispersion of tissue excitability. The "critical point hypothesis" provides the minimum topology required for this type of reentry; the "graded response hypothesis" can be viewed as providing a more detailed explanation of how this topology is actually realized. Triggering mechanisms due to the "break" mode of stimulation also posits a new mechanism for defibrillation.

Animals↗

Kaposi's sarcoma-associated herpesvirus encodes a bZIP protein with homology to BZLF1 of Epstein-Barr virus.

Kaposi's sarcoma-associated herpesvirus (KSHV) is a recently discovered human gamma herpesvirus strongly implicated in AIDS-related neoplasms. We report here the identification in the KSHV genome of a gene for a protein designated K-bZIP and belonging to the basic-leucine zipper (bZIP) family of transcription factors. K-bZIP shows significant homology to BZLF1, which plays a key role in the replication and reactivation of Epstein-Barr virus. K-bZIP is a homodimerizing protein of 237 amino acids with a prototypic bZIP domain at the C terminus. The N-terminal portion of K-bZIP is derived from the K8 open reading frame which, through in-frame splicing, adjoins the ZIP domain. This structure was revealed by rapid analysis of cDNA ends, followed by cloning of the entire cDNA. A 1.35-kb transcript encoding K-bZIP was detected in BCBL-1 cells treated with 12-O-tetradecanoylphorbol-13-acetate. The synthesis of this transcript was blocked by the protein synthesis inhibitor cycloheximide but not by the viral DNA synthesis inhibitor phosphonoacetate, a result which classifies it as an early lytic gene. RNase protection analysis further mapped the major transcription start site for the 1.35-kb K-bZIP mRNA and identified two other splice variants which encode proteins with the N-terminal portion of K-bZIP but lacking the C-terminal ZIP domain. Full-length K-bZIP forms dimers with itself, and the C terminus encompassing the ZIP domain is required for this process. Our studies set the stage for understanding the role of K-bZIP in the replication and reactivation of the KSHV genome.

Amino Acid Sequence↗