Biomedical subjects

Y L Lin

Publications and source records attributed to Y L Lin.

At least 73 records · Page 4Linked to original sources

PubMed1999

Prevalence of microalbuminuria and relationship to the risk of cardiovascular disease in the Japanese population.

The prevalence of microalbuminuria and its relationship to cardiovascular disease risk factors were examined in subjects participating in an annual physical and laboratory examination program. The urinary albumin concentration and the urinary albumin/creatinine ratio were determined in morning urine specimens. A turbidimetric immunoassay was used for the measurement of urinary albumin. Of the 731 subjects, 41 (5.6%) who were weakly positive or positive on a routine dipstick test for protein were excluded from the final analysis of data. Microalbuminuria was present in 14.5% of the men, in 12.4% of the women, and in 13.2% of the entire subject population when defined as a urinary albumin concentration of 30-299 microgram/ml. The prevalence of microalbuminuria was significantly higher in subjects with a high normal blood pressure (15.0%) or hypertension (26.2%) as compared with normotensive subjects (6.5%). Subjects with impaired glucose tolerance (24.3%) or hyperglycemic subjects (50.0%) had a significantly higher prevalence of microalbuminuria than normoglycemic subjects (11.3%). The prevalence of microalbuminuria was significantly higher in subjects with left ventricular hypertrophy (47.1%) as compared with those with normal electrocardiograms (11.3%). A good correlation was observed between urinary albumin concentration and albumin/creatinine ratio, and both showed a significant positive correlation with age, systolic and diastolic blood pressures, and fasting plasma glucose, total serum protein, albumin, and triglyceride levels, but not with angiotensin-converting enzyme activity. Multiple regression analysis demonstrated that both the urinary albumin concentration and the albumin/creatinine ratio show a significant positive correlation with systolic blood pressure and fasting plasma glucose. The prevalence of microalbuminuria was about 13% in this Japanese cohort, and the systolic blood pressure and the fasting plasma glucose level were demonstrated as independent risk indicators for both urinary microalbumin level and urinary microalbumin/creatinine ratio.

Adult↗

PubMed1999

Signaling pathway and pepsinogen secretion in Helicobacter pylori-infected human gastric adenocarcinoma.

Although in vitro studies have suggested that Helicobacter pylori not only attaches to cultured cells but also induces signal transduction events in host cells, the underlying mechanism of H. pylori action has yet to be fully investigated. In the present study, a cytotoxin-positive H. pylori was used to infect and examined for its effect on the stimulation of second messengers in human gastric adenocarcinoma (AGS). Results showed that H. pylori increased cytosolic free calcium concentration [Ca2+]i in host cells in a dose-dependent manner. The increase of [Ca2+]i was due to release from the intracellular Ca2+ store as well as entry to the extracellular Ca2+. H. pylori infection on host cells was also found to induce the generations of inositol phosphates, adenosine 3', 5'-cyclic monophosphate, and guanosine 3',5'-cyclic monophosphate, and to stimulate the secretion of pepsinogen.

Adenocarcinoma↗

PubMed1999

Association between serotonin transporter gene polymorphism and smoking among Japanese males.

The serotoninergic system may be involved in smoking behavior because nicotine increases brain serotonin secretion, nicotine withdrawal decreases serotonin levels, and a selective serotonin reuptake inhibitor antagonizes the response to nicotine. Compared with the L allele, the S allele of the polymorphism in the upstream regulatory region of the serotonin transporter gene is associated with decreased transcription efficiency of the 5-HTT gene promoter. We examined this polymorphism in a Japanese population consisting of 387 males from two different areas in Japan. The L allele was observed significantly more often in smokers (21%) than in nonsmokers (lifetime nonsmokers + ex-smokers, 14%; P = 0.005). The presence of the L allele (the L/L + L/S genotypes) was also significantly increased in smokers (37%) compared with that in nonsmokers (24%; P = 0.003). The present study suggests that individuals with the S/S genotype are less inclined to smoke and/or can more easily stop smoking than others, supporting a role of the serotoninergic system in smoking behavior.

Adult↗

PubMed1998

An eukaryotic RuvB-like protein (RUVBL1) essential for growth.

A human protein (RUVBL1), consisting of 456 amino acids (50 kDa) and highly homologous to RuvB, was identified by using the 14-kDa subunit of replication protein A (hsRPA3) as bait in a yeast two-hybrid system. RuvB is a bacterial protein involved in genetic recombination that bears structural similarity to subunits of the RF-C clamp loader family of proteins. Fluorescence in situ hybridization analysis demonstrated that the RUVBL1 gene is located at 3q21, a region with frequent rearrangements in different types of leukemia and solid tumors. RUVBL1 co-immunoprecipitated with at least three other unidentified cellular proteins and was detected in the RNA polymerase II holoenzyme complex purified over multiple chromatographic steps. In addition, two yeast homologs, scRUVBL1 and scRUVBL2 with 70 and 42% identity to RUVBL1, respectively, were revealed by screening the complete Saccharomyces cerevisiae genome sequence. Yeast with a null mutation in scRUVBL1 was nonviable. Thus RUVBL1 is an eukaryotic member of the RuvB/clamp loader family of structurally related proteins from bacteria and eukaryotes that is essential for viability of yeast.

ATPases Associated with Diverse Cellular Activitie↗

PubMed1998

Phosphatidylinositol 3-kinase is required for the regulation of hepatitis B surface antigen production and mitogen-activated protein kinase activation by insulin but not by TPA.

Insulin suppresses hepatitis B surface antigen (HBsAg) gene expression and stimulates cell proliferation in human hepatoma Hep3B cells. 12-O-tetradecanoyl phorbol-13-acetate, TPA, has been demonstrated to mimic insulin actions in these cells. We examined the role of phosphatidylinositol 3-kinase (PI 3-kinase) in the signaling pathways of insulin and TPA towards these two biological phenomena in Hep3B cells. The pre-treatment of 5 microM of wortmannin diminished insulin suppressed HBsAg production and completely abolished insulin stimulated cell proliferation. However, wortmannin had no effect on TPA actions in both HBsAg suppression and cell growth stimulation. We further investigated the effect of wortmannin in mitogen-activated protein kinases (MAPKs) activation induced by insulin or TPA. After the pretreatment of wortmannin, insulin activated MAPKs was completely blocked, but TPA was still capable to activate MAPKs. These results suggest that PI 3-kinase is involved in insulin actions but not in TPA effects, and allow us to dissociate the signaling pathways of insulin and TPA in human hepatoma Hep3B cells.

Androstadienes↗

PubMed1998

In vitro stimulation of C1s proteolytic activities by C1s-presenting autoantibodies from patients with systemic lupus erythematosus.

Anti-C1s autoantibodies (IgG forms), which recognize the conjunction of C1s heavy chain and light chain (C1s-presenting autoantibodies) from patients with systemic lupus erythematosus (SLE), have been found to stimulate C1s enzymatic activities. This is due to acceleration of the proteolytic hydrolysis of the synthetic substrate C1-1 by C1s, enhancement of the complex formation of C1s with its natural pseudosubstrate, C1 inhibitor (C1 inh), and promotion of proteolytic activation of its natural substrate, C4. Seven of fifteen samples from patients with SLE were found to contain such autoantibodies. The hydrolysis of the synthetic substrate C1-1 catalyzed by C1s in 25 to 27 min in the presence of anti-C1s autoantibodies was equivalent to the hydrolysis of C1-1 catalyzed by C1s alone or C1s with control IgG from healthy sera in 110 min, approximately fourfold faster than the reaction in the absence of anti-C1s autoantibodies. Densitometry scanning data showed that the formation of the C1s-C1 inh complex in the presence of anti-C1s autoantibodies was three to four times greater than that with control IgG. It was also noticed that the autoantibodies convert almost all of the latent forms of C1s to an active form that binds to C1 inh. Another group of Western blots showed that C1s cleaved C4 alpha-chain three times faster in the presence of autoantibodies than of control IgG. It is likely that the overconsumption of complement components is common in the pathogenesis of tissue damage occurring in SLE.

Antigen-Antibody Complex↗

PubMed1998

Taxol induces concomitant hyperphosphorylation and reorganization of vimentin intermediate filaments in 9L rat brain tumor cells.

Taxol, a microtubule stabilizing agent, has been extensively investigated for its antitumor activity. The cytotoxic effect of taxol is generally attributed to its antimicrotubule activity and is believed to be cell cycle dependent. Herein, we report that taxol induces hyperphosphorylation and reorganization of the vimentin intermediate filament in 9L rat brain tumor cells, in concentration- and time-dependent manner. Phosphorylation of vimentin was maximum at 10(-6) M of taxol treatment for 8 h and diminished at higher (10(-5) M) concentration. Enhanced phosphorylation of vimentin was detectable at 2 h treatment with 10(-6) M taxol and was maximum after 12 h of treatment. Taxol-induced phosphorylation of vimentin was largely abolished in cells pretreated with staurosporine and bisindolymaleimide but was unaffected by H-89, KT-5926, SB203580, genistein, and olomoucine. Thus, protein kinase C may be involved in this process. Hyperphosphorylation of vimentin was accompanied by rounding up of cells as revealed by scanning electron microscopy. Moreover, there was a concomitant reorganization of the vimentin intermediate filament in the taxol-treated cells, whereas the microtubules and the actin microfilaments were less affected. Taken together, our data demonstrate that taxol induces hyperphosphorylation of vimentin with concomitant reorganization of the vimentin intermediate filament and that this process may be mediated via a protein kinase C signaling pathway.

Animals↗

PubMed1998

Identification of neutral and acidic sphingomyelinases in Helicobacter pylori.

We demonstrated for the first time the presence of sphingomyelinase (SMase) in Helicobacter pylori. Activation of SMase has been implicated as the cause of elevation of cellular ceramide levels and consequently of apoptosis. The data indicate that there are two classes of SMase, defined by their optimal pHs and cellular locations, existing in H. pylori. One is an Mg(2+)-dependent membrane-bound enzyme with an optimal activity at pH 7, and the other is an Mg(2+)-independent cytosolic enzyme with an optimal activity at pH 5. Bisalumin, a bismuth salt, was found to inhibit the activities of both forms of SMase regardless of the presence of Mg2+. By Western blot analysis, the membrane-bound SMases of H. pylori and Bacillus cereus were shown to be antigenically related and to have a similar denatured molecular mass of 28 kDa.

Bismuth↗

PubMed1998

The evolutionarily conserved zinc finger motif in the largest subunit of human replication protein A is required for DNA replication and mismatch repair but not for nucleotide excision repair.

The largest subunit of the replication protein A (RPA) contains an evolutionarily conserved zinc finger motif that lies outside of the domains required for binding to single-stranded DNA or forming the RPA holocomplex. In previous studies, we showed that a point mutation in this motif (RPAm) cannot support SV40 DNA replication. We have now investigated the role of this motif in several steps of DNA replication and in two DNA repair pathways. RPAm associates with T antigen, assists the unwinding of double-stranded DNA at an origin of replication, stimulates DNA polymerases alpha and delta, and supports the formation of the initial short Okazaki fragments. However, the synthesis of a leading strand and later Okazaki fragments is impaired. In contrast, RPAm can function well during the incision step of nucleotide excision repair and in a full repair synthesis reaction, with either UV-damaged or cisplatin-adducted DNA. Two deletion mutants of the Rpa1 subunit (eliminating amino acids 1-278 or 222-411) were not functional in nucleotide excision repair. We report for the first time that wild type RPA is required for a mismatch repair reaction in vitro. Neither the deletion mutants nor RPAm can support this reaction. Therefore, the zinc finger of the largest subunit of RPA is required for a function that is essential for DNA replication and mismatch repair but not for nucleotide excision repair.

Binding Sites↗

PubMed1998

Localizations of NS3 and E proteins in mouse brain infected with mutant strain of Japanese encephalitis virus.

Infection with a mutant Japanese encephalitis virus (JEV) strain RP-2ms showed reduced neurovirulence than wild type or RP-9 strains after inoculation in BALB/c mice. However, higher intracellular viral titer was detected in Rp-2ms infected cultured cells. Localizations of non-structural 3 (NS3) and envelope (E) proteins were demonstrated by immunocytochemistry. NS3 protein was primarily found in the pyramidal neurons in cerebrum, in the molecular and granular layers of cerebellum. Neither E nor NS3 protein was detected in Purkinje cells of the cerebellum. Immunoelectron microscopic observations showed that E and NS3 proteins were positive in JEV-induced membranous systems, mainly hypertrophic rough endoplasmic reticulum (rER) and membrane vesicle structure (MVS) but not smooth membrane structure. Virus particles were seen in the Golgi apparatus, rER, nuclear envelope, MVS and cytoplasmic vacuoles. Different mechanisms of intracellular trapping in vivo provide a possible basis for attenuation of RP-2ms strains of JEV.

Animals↗

PubMed1998

Comparison of the immunogenicity of wasp venom peptides with or without carbohydrate moieties.

Three synthetic vespulakinin analogues either with or without carbohydrate moieties and mastoparan B isolated from Vespa basalis venom were investigated for their immunogenic activity and solution conformation. Mice immunized with these wasp venom peptides, with the exception of (Gal alpha)Thr3, (Gal alpha)Thr4-vespulakinin 1, showed positive antibody responses. However, the response elicited by mastoparan B was much higher than those induced by vespulakinin analogues. The class of antibody induced by these peptides was identified as an IgG1 isotype with kappa-light chain, suggesting stimulation of a T-cell-dependent immune response by these peptides. According to the circular dichroism spectra of these peptides, the structures of the vespulakinin analogues in solution were largely unordered, while mastoparan B exhibited a conformation rich in alpha-helices. The presence of carbohydrate moieties and the rather random structure in vespulakinins may interfere with T-cell recognition of the peptides, leading to lower immune responses.

Allergens↗

PubMed1998

Structures and biological activities of new wasp venom peptides isolated from the black-bellied hornet (Vespa basalis) venom.

The black-bellied hornet (Vespa basalis) is the most dangerous species of vespine wasps found in Taiwan. The hornet possesses a highly toxic venom which is rich in toxin, enzymes and biologically active peptides. Using ultrafiltration to remove the high molecular weight toxin and enzymes followed by reverse-phase HPLC, three bioactive tridecapeptides, designated "HP-1, HP-2 and HP-3" were isolated from the venom. Their amino acid sequences were determined as: HP-1: LFRLIAKTLGSLM, HP-2: LFRLLANTLGKIL, HP-3: IFGLLAKTLGNLF. The primary structures of these peptides appear to be homologous to those of chemotactic peptides isolated from other vespid venoms. However, these peptides show little chemotactic activity on human neutrophils and have distinct tripeptide sequences at the amino and carboxylic terminal sides, as compared with other hornet chemotactic peptides. The lack of Pro3 which is a characteristic structure of vespid chemotactic peptides in their sequences is most distinctive. Circular dichroism spectra of these peptides measured in 20% trifluoroethanol show a high content of alpha-helical conformation. All three peptides provoked local edema in rat hind paw, which could be inhibited by antihistamine (diphenhydramine) and drug with antiserotonin activity (cyproheptadine). These peptides also exhibited a potent hemolytic activity which was potentiated by a non-lytic dose of the hornet lethal protein, suggesting a supporting role of these peptides in the lethal effect of Vespa basalis venom.

Amino Acid Sequence↗

PubMed1998

Thrombin-induced DNA synthesis of cultured human dental pulp cells is dependent on its proteolytic activity and modulated by prostaglandin E2.

To clarify the roles of alpha-thrombin and prostaglandin E2 (PGE2) in the healing and inflammatory processes of dental pulp, their effects on the DNA synthesis of human pulp cells were investigated by measurement of [3H]thymidine incorporation. At a concentration range of 1 to 25 units/ml, alpha-thrombin stimulated DNA synthesis of the pulp cells by 1.5 to 2.6-fold. On the contrary, PGE2 (> 0.05 microgram/ml) suppressed DNA synthesis by 24 to 39%. Using reverse transcriptase-polymerase chain reaction, thrombin receptor mRNA expression was identified in the pulp cells. Furthermore, alpha-thrombin-induced DNA synthesis could be inhibited by antithrombin III (2 units/ml) with heparin (2 units/ml) or D-Phe-Pro-ArgCH2Cl (50 micrograms/ml). PGE2 (0.1 to 0.5 microgram/ml) also inhibited the thrombin-induced DNA synthesis by 39 to 64%. These results imply that pulp cells express the thrombin receptor that is activated by the serine protease activity of thrombin. Interactions of thrombin and PGE2 are important in modulating the inflammatory and healing processes of the pulp.

Cells, Cultured↗

PubMed1998

Promotional effect of N-nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (a nitrosated Maillard reaction product) in mouse fibroblast cells.

N-Nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (NO-NTA) is a product of model browning system generated in the presence of sodium nitrite. The chemical structure of this compound has been confirmed by UV, mass and nuclear magnetic resonance, and infrared spectroscopy in our previous study. A two-stage transformation protocol was used to chemically transform the mouse embryo fibroblasts C3H10T1/2 cells. To initiate transformation, the cells were treated with benzo[a]pyrene (BaP) (0.1 mg/ml), and NO-NTA (0.01, 0.1 and 1 mg/ml) was employed subsequently to complete the transformation process. Malignant transformed foci were formed in BaP-initiated and NO-NTA promoted C3H10T1/2 cells after 8 wk. Cells treated with NO-NTA alone failed to induce transformation. However, cells initiated with BaP and promoted by cells initiated with BaP and promoted by NO-NTA demonstrated oncogenic properties. Cell lines transformed with NO-NTA-transformed colonies exhibited enhanced growth rate, anchorage independence and tumorigenicity in animals relative to parent cells. These results indicate that NO-NTA is a new tumour promoter and may induce tumour promotion by two-stage oncogenesis. Further studies on the mechanism of action of NO-NTA are now in progress.

Animals↗

PubMed1998

The impact of National Health Insurance on the volume and severity of emergency department use.

National Health Insurance (NHI) was introduced in Taiwan on March 1, 1995. To evaluate the Influence of NHI on the volume and severity of emergency department (ED) visits, a retrospective analysis was conducted. The results showed that NHI did not result in a substantial increase in ED volume (P > .05), but the severity as measured by triage nurses did increase (P < .01). There was no change in the number of admissions to the intensive care units of patients transferred from smaller hospitals, but its percentage significantly decreased (P < .05), which may be a result of the significant increase in the total number of interhospital transfers. It was hypothesized that the universal coverage and a copayment contributed to these changes.

Emergency Service, Hospital↗

PubMed1998

Four new sesquiterpenes from Petasites formosanus.

Four new eremophilane-type sesquiterpenes, petasones A (1) and B (2), S-petasitin (3), and petasinol (4), were isolated from the aerial parts of Petasites formosanus. Their structures were elucidated by spectroscopic and chemical methods as 3alpha-[(Z)-3-methylsulfinylacryloxy]eremophila-7(11), 9-dien-8-one; 3alpha-[(E)-3-methylsulfinylacryloxy]eremophila-7(11), 9-dien-8-one; 3alpha-[(Z)-3-methylthioacryloxy]-11-hydroxyeremophila -6, 9-dien-8-one; and 3alpha-[(E)-3-methylsulfinylacryloxy]-8beta-hydroxy eremophila-9, 11-diene, respectively.

Carbohydrate Sequence↗

PubMed1998

DNA immunization with Japanese encephalitis virus nonstructural protein NS1 elicits protective immunity in mice.

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is a zoonotic pathogen that is prevalent in some Southeast Asian countries and causes acute encephalitis in humans. To evaluate the potential application of gene immunization to JEV infection, we characterized the immune responses from mice intramuscularly injected with plasmid DNA encoding JEV glycoproteins, including the precursor membrane (prM) plus envelope (E) proteins and the nonstructural protein NS1. When injected with the plasmid expressing prM plus E, 70% of the immunized mice survived after a lethal JEV challenge, whereas when immunized with the plasmid expressing NS1, 90% of the mice survived after a lethal challenge. As a control, the mice immunized with the DNA vector pcDNA3 showed a low level (40%) of protection, suggesting a nonspecific adjuvant effect of the plasmid DNA. Despite having no detectable neutralizing activity, the NS1 immunization elicited a strong antibody response exhibiting cytolytic activity against JEV-infected cells in a complement-dependent manner. By contrast, immunization with a construct expressing a longer NS1 protein (NS1'), containing an extra 60-amino-acid portion from the N terminus of NS2A, failed to protect mice against a lethal challenge. Biochemical analyses revealed that when individually expressed, NS1 but not NS1' could be readily secreted as a homodimer in large quantity and could also be efficiently expressed on the cell surface. Interestingly, when NS1 and NS1' coexisted in cells, the level of NS1 cell surface expression was much lower than that in cells expressing NS1 alone. These data imply that the presence of partial NS2A might have a negative influence on an NS1-based DNA vaccine. The results herein clearly illustrate that immunization with DNA expressing NS1 alone is sufficient to protect mice against a lethal JEV challenge.

Animals↗

PubMed1998

Study of Dengue virus infection in SCID mice engrafted with human K562 cells.

Here we report that severe combined immunodeficient (SCID) mice engrafted with human K562 cells (K562-SCID mice) can be used as an animal model to study dengue virus (DEN) infection. After intratumor injection into K562 cell masses of PL046, a Taiwanese DEN-2 human isolate, the K562-SCID mice showed neurological signs of paralysis and died at approximately 2 weeks postinfection. In addition to being detected in the tumor masses, high virus titers were detected in the peripheral blood and the brain tissues, indicating that DEN had replicated in the infected K562-SCID mice. In contrast, the SCID mice were resistant to DEN infection and the mock-infected K562-SCID mice survived for over 3 months. These data illustrate that DEN infection contributed directly to the deaths of the infected K562-SCID mice. Other serotypes of DEN were also used to infect the K562-SCID mice, and the mortality rates of the infected mice varied with different challenge strains, suggesting the existence of diverse degrees of virulence among DENs. To determine whether a neutralizing antibody against DEN in vitro was also protective in vivo, the K562-SCID mice were challenged with DEN-2 and received antibody administration at the same time or 1 day earlier. Our results revealed that the antibody-treated mice exhibited a reduction in mortality and a delay of paralysis onset after DEN infection. In contrast to K562-SCID, the persistently DEN-infected K562 cells generated in vitro invariably failed to be implanted in the mice. It seems that in the early stage of implantation, a gamma interferon activated, nitric oxide-mediated anti-DEN effect might play a role in the innate immunity against DEN-infected cells. The system described herein offers an opportunity to explore DEN replication in vivo and to test various antiviral protocols in infected hosts.

Animals↗