Biomedical subjects

L L Lin

Publications and source records attributed to L L Lin.

At least 37 records · Page 2Linked to original sources

PubMed1999

Engagement of tumor necrosis factor (TNF) receptor 1 leads to ATF-2- and p38 mitogen-activated protein kinase-dependent TNF-alpha gene expression.

Engagement of the tumor necrosis factor-alpha (TNF-alpha) receptors by the TNF-alpha ligand results in the rapid induction of TNF-alpha gene expression. The study presented here shows that autoregulation of TNF-alpha gene transcription by selective signaling through tumor necrosis factor receptor 1 (TNFR1) requires p38 mitogen-activated protein (MAP) kinase activity and the binding of the transcription factors ATF-2 and Jun to the TNF-alpha cAMP-response element (CRE) promoter element. Consistent with these findings, TNFR1 engagement results in increased p38 MAP kinase activity and p38-dependent phosphorylation of ATF-2. Furthermore, overexpression of MADD (MAP kinase-activating death domain protein), an adapter protein that binds to the death domain of TNFR1 and activates MAP kinase cascades, results in CRE-dependent induction of TNF-alpha gene expression. Thus, the TNF-alpha CRE site is the target of TNFR1 stimulation and mediates the autoregulation of TNF-alpha gene transcription.

Activating Transcription Factor 2↗

PubMed1999

Activation of the erythropoietin receptor is not required for internalization of bound erythropoietin.

Erythropoietin (EPO) is required for the survival and expansion of red blood cell progenitor cells and supports continued differentiation of these committed progenitors to mature red blood cells. After binding to its cognate receptor, EPO promotes receptor homodimerization, activation of receptor-associated JAK2, subsequent receptor tyrosine phosphorylation, and transduction of signal. EPO is also internalized and degraded in lysosomes. The contribution of EPO-induced receptor internalization to modulation of EPO signals has not been determined. To examine this question, we generated a panel of hematopoietic cell lines containing progressively truncated isoforms of the erythropoietin receptor (EPO-R) and determined the rate and extent of EPO internalization and receptor downregulation. We demonstrated that a membrane-proximal domain of the cytoplasmic tail of the EPO-R was the minimal region required for EPO-induced receptor internalization. This cytoplasmic domain is also the minimal domain required for activation of JAK2, a cytosolic tyrosine kinase essential for the function of the EPO-R. However, neither EPO activation of cytosolic JAK2 tyrosine kinase activity nor tyrosine phosphorylation of the EPO-R cytoplasmic tail was required for EPO-induced receptor downregulation. Both functional and nonfunctional cell surface receptor isoforms were internalized equally. These results suggest that, for downregulation of cell surface ligand occupied EPO-R and possibly for signaling receptors of the cytokine receptor superfamily in general, internalization of cell surface ligand occupied receptors may follow a pathway distinct from signaling receptors of the receptor tyrosine kinase (RTK) family.

Animals↗

PubMed1999

The role of a conserved histidine residue, His324, in Trigonopsis variabilis D-amino acid oxidase.

To investigate the functional role of an invariant histidine residue in Trigonopsis variabilis D-amino acid oxidase (DAAO), a set of mutant enzymes with replacement of the histidine residue at position 324 was constructed and their enzymatic properties were examined. Wild-type and mutant enzymes have been purified to homogeneity using the His-bound column and the molecular masses were determined to be 39.2 kDa. Western blot analysis revealed that the in vivo synthesized mutant enzymes are immuno-identical with that of the wild-type DAAO. The His324Asn and His324Gln mutants displayed comparable enzymatic activity to that of the wild-type enzyme, while the other mutant DAAOs showed markedly decreased or no detectable activity. The mutants, His324/Asn/Gln/Ala/Tyr/Glu, exhibited 38-181% increase in Km and a 2-10-fold reduction in kcat/Km. Based on the crystal structure of a homologous protein, pig kidney DAAO, it is suggested that His324 might play a structural role for proper catalytic function of T. variabilis DAAO.

Amino Acid Sequence↗

PubMed1999

Protein kinase C epsilon-dependent pathway of extracellular signal-regulated protein kinase activation by P2Y1 and P2Y2 purinoceptors that activate cytosolic phospholipase A2 in endothelial cells.

The aim of this study was to investigate the stimulating effects on arachidonic acid release of P2Y1 and P2Y2 receptor-selective agonists, 2-methylthio-ATP (2MeSATP) and UTP, respectively, in bovine pulmonary artery endothelial cells. Exposure of cells to 2MeSATP and UTP led to the release of arachidonic acid, a response which was abolished by the removal of extracellular Ca2+ and methyl arachidonyl fluorophosphonate. Phorbol 12-myristate 13-acetate (PMA) itself not only stimulated arachidonic acid release but also played a permissive role in the response to UTP. However, PMA failed to enhance the arachidonic acid response induced by 2MeSATP, probably due to greater attenuation of the [Ca2+]i increase caused by 2MeSATP than UTP. Inhibition of protein kinase C with Ro 31-8220 (1-[3-(amidinothio) propyl-1H-indoyl-3-yl]-3-(1-methyl-1H-indoyl-3-yl)-maleimide -methane sulphate) and staurosporine, but not with Go 6976 (12-(-2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-indolo(2, 3-a)pyrrolo(3,4-c)carbazole), reduced the arachidonic acid response of 2MeSATP, UTP and PMA. PMA-induced potentiation of the UTP response reached a maximum after a 1-h preincubation, then declined and eventually lost its effect when the preincubation lasted up to 8 h. Among the protein kinase C isoforms present in endothelial cells, betaI and epsilon could be down-regulated by treatment with PMA for 4-24 h. PD 098059 (2-(2-Amino-3-methoxyphenyl)-4H-1-benzopyran-4-one) inhibited extracellular signal-regulated protein kinase activation, cytosolic phospholipase A2 phosphorylation and arachidonic acid release caused by 2MeSATP, UTP and PMA. Taken together, our results demonstrate that P2Y1 and P2Y2 purinoceptors mediate arachidonic acid release by activating cytosolic phospholipase A2 through an elevation of [Ca2+]i and protein kinase C epsilon-, extracellular signal-regulated protein kinase-dependent phosphorylation.

Adenosine Triphosphate↗

PubMed1999

Antimicrobial activity of tea as affected by the degree of fermentation and manufacturing season.

Bacillus subtilis, Escherichia coli, Proteus vulgaris, Pseudomonas fluorescens, Salmonella sp. and Staphylococcus aureus were used to test the antimicrobial activity of tea flush extract and extracts of various tea products. Among the six test organisms, P. fluorescens was the most sensitive to the extracts, while B. subtilis was the least sensitive. In general, antimicrobial activity decreased when the extents of tea fermentation increased. The antimicrobial activities of tea flush extract and extracts of tea products with different extents of fermentation varied with test organisms. Tea flush and Green tea, the unfermented tea, exerted the strongest antimicrobial activity followed by the partially fermented tea products such as Longjing, Tieh-Kuan-Ying, Paochung, and Oolong teas. On the other hand, Black tea, the completely fermented tea, showed the least antimicrobial activity. It was also noted that extracts of Oolong tea prepared in summer exhibited the strongest antimicrobial activity, followed by those prepared in spring, winter and fall.

Anti-Bacterial Agents↗

PubMed1999

Evaluation of the Strep B OIA test compared to standard culture methods for detection of group B streptococci.

OBJECTIVE: This study evaluated the accuracy of the commercial product Strep B OIA (optical immunoassay) compared to the standard agar and broth culture methods for detecting vaginal colonization with group B streptococcus (GBS). METHODS: Preoperative vaginal cultures were obtained from 141 nonpregnant gynecological patients undergoing major gynecologic surgery. Major gynecologic surgery was defined as benign gynecologic, gyne-oncology, and urogynecologic procedures. The results of the Strep B OIA test were compared to the results obtained from SXT agar (selective for GBS), colistin-nalidixic acid ((CNA) agar, and Todd-Hewitt broth cultures. RESULTS: The prevalence of vaginal GBS colonization in this population was 20.6%. The sample sensitivity and specificity of the OIA method were 58.6% and 85.7%, respectively. These values are lower than the sensitivity and specificity of 85.4% and 91.5%, respectively, given in the OIA package insert. Although the sample negative predictive value was fairly high (88.9%), the positive predictive value was low (51.5%). CONCLUSION: Although a previous study stated that the product Strep B OIA reduces the time required to obtain results (30 minutes versus days) and can, therefore, function as a useful diagnostic tool in the management of early-onset GBS disease, the present study's finding of low sensitivity and low positive predictive value indicates that this test may have very limited clinical value.

Adult↗

PubMed1999

Effects of different concentrations of atropine on controlling myopia in myopic children.

Although 1% atropine effectively slows myopia progression, it is associated with adverse effects, including photophobia, blurred near vision, and poor compliance. We investigated whether lower doses of atropine would control myopia progression. One hundred and eighty-six children, from 6 to 13 years of age, were treated each night with different concentrations of atropine eye drops or a control treatment for up to 2 years. The mean myopic progression in each of the groups was 0.04 +/-0.63 diopter per year (D/Y) in the 0.5% atropine group, 0.45+/-0.55 D/Y in the 0.25% atropine group, and 0.47+/-0.91 D/Y in the 0.1% atropine group. All atropine groups showed significantly less myopic progression than the control group (1.06+/-0.61 D/Y) (p<0.01). Our study also showed that 61% of students in the 0.5% atropine group, 49% in the 0.25% atropine group and 42% in the 0.1% atropine group had no myopic progression. However, 4% of children in the 0.5% atropine group, 17% in the 0.25% atropine group, and 33% in the 0.1% atropine group still had fast myopic progression (>-1.0 D/Y). In contrast, only 8% of the control group showed no myopic progression and 44% had fast myopic progression. These results suggest that all three concentrations of atropine had significant effects on controlling myopia; however, treatment with 0.5% atropine was the most effective.

Adolescent↗

PubMed1999

Epidemiologic study of ocular refraction among schoolchildren in Taiwan in 1995.

PURPOSE: In order to understand and update the prevalence of myopia in Taiwan, a nationwide survey was performed in 1995. METHODS: We stratified the cluster sampling by developmental grading of the city, using a size proportional to the population. Two cities were randomly selected from each city grading. The total number of students enrolled was 11,178, including 5,676 boys and 5,502 girls. The refractive status and corneal radius of each student were measured with an autorefractometer under cycloplegia and checked with retinoscopy. Axial length was measured with biometric ultrasound. RESULTS: The myopic rate was from 12% at the age of 6, it increased to 56% at the age of 12, and then to 76% at the age of 15. A myopic rate of 84% was found for the age range of 16 to 18. The prevalence of high myopia (over -6.0 D) at the age of 18 was 20% in girls and 12% in boys. The mean refractive status became myopic at the age of 9, then increased to -3.92 D in girls and -2.71 D in boys at the age of 18. The increase of axial length is correspondent with the progression of myopia. The anterior chamber depth (ACD) was deeper with age and the severity of myopia, whereas the corneal curvature remained unchanged. The lens thickness became thinner from age 7 to 13, then it became thicker with age and the severity of myopia after age 15. The prevalence and degree of myopia in girls was more severe than in boys. CONCLUSIONS: The prevalence of myopia in Taiwan increased year by year. The increase in severity and prevalence of high myopia may be due to earlier onset.

Adolescent↗

PubMed1998

A conserved aspartate is essential for FAD binding and catalysis in the D-amino acid oxidase from Trigonopsis variabilis.

To evaluate the possible contribution of Asp206 of Trigonopsis variabilis D-amino acid oxidase (DAO) to its flavin adenine dinucleotide (FAD) binding and catalytic function, six mutant enzymes were constructed by site-directed mutagenesis. Western immunoblot analysis revealed that a protein with an apparent molecular mass of about 39.2 kDa was present in the cell-free extracts of wild-type and mutant strains. Replacement of Asp206 with Leu, Gly, and Asn resulted in the loss of DAO activity and characteristic absorption spectrum for flavoenzyme, while the other mutant DAOs, Asp206Glu, Asp206Ser, and Asp206Ala, exhibited a similar spectral profile to that of wild-type enzyme and retained about 6-90% of the enzyme activity. These results suggested that Asp206 of T. variahilis DAO might play an important role in the binding of FAD.

Amino Acid Sequence↗

PubMed1998

A novel calcium-independent phospholipase A2, cPLA2-gamma, that is prenylated and contains homology to cPLA2.

We report the cloning and characterization of a novel membrane-bound, calcium-independent PLA2, named cPLA2-gamma. The sequence encodes a 541-amino acid protein containing a domain with significant homology to the catalytic domain of the 85-kDa cPLA2 (cPLA2-alpha). cPLA2-gamma does not contain the regulatory calcium-dependent lipid binding (CaLB) domain found in cPLA2-alpha. However, cPLA2-gamma does contain two consensus motifs for lipid modification, a prenylation motif (-CCLA) at the C terminus and a myristoylation site at the N terminus. We present evidence that the isoprenoid precursor [3H]mevalonolactone is incorporated into the prenylation motif of cPLA2-gamma. Interestingly, cPLA2-gamma demonstrates a preference for arachidonic acid at the sn-2 position of phosphatidylcholine as compared with palmitic acid. cPLA2-gamma encodes a 3-kilobase message, which is highly expressed in heart and skeletal muscle, suggesting a specific role in these tissues. Identification of cPLA2-gamma reveals a newly defined family of phospholipases A2 with homology to cPLA2-alpha.

Amino Acid Sequence↗

PubMed1998

A case-control study on risk factors for Type 1 diabetes in Taipei City.

To search for non-genetic risk factors for Type 1 diabetes among Chinese, a total of 117 Type 1 diabetes patients identified from the Taipei Type 1 Diabetes Registry and 193 frequency-matched controls with normal glucose tolerance were recruited. The controls were selected from classmates or colleagues of cases with similar distributions of age, sex, and parental and individual educational levels. Information on Type 1 diabetes risk factors was obtained through standardized interviews according to a structured questionnaire administered by two trained nurses. In the univariate analyses, the order of pregnancy of the study subject, the number of spontaneous and induced abortions reported by the mother before giving birth to the study subjects, the duration of breast-feeding and monthly family income were significantly associated with Type 1 diabetes in a dose-response manner. There was a U-shaped relationship with Type 1 diabetes for birth weight and paternal age at the conception of the study subjects. After further adjustment for multiple risk factors through logistic regression analysis, the biological gradient for pregnancy order, duration of breast-feeding, and the U-shaped relationship for paternal age remained statistically significant.

Abortion, Induced↗

PubMed1998

The cycloplegic effects of cyclopentolate and tropicamide on myopic children.

Thirty-seven myopic children were given either 1-2 drops of 1% cyclopentolate or 1% tropicamide twice with 5 min intervals to evaluate the time course and maximal cycloplegic effect of both agents. The other fifteen subjects were given 1% tropicamide initially, then 1% cyclopentolate given after 30 min of maximal effect of tropicamide appeared to evaluate whether the effect of cyclopentolate was superior to tropicamide. Cycloplegic refraction was measured with an auto-refractometer (Topcon RK-3000) before drug delivery and every 15 min thereafter, for 90 min. The maximal cycloplegic effect of cyclopentolate was around 45 min, then it remained stable until 90 min after the last instillation. The effect of tropicamide was faster than that of cyclopentolate. It was around 30 min, then it stabilized until 75 min. The extra effect of cyclopentolate over tropicamide was minimal (only -0.1D). The power of cornea and astigmatism were not affected by either agent. However, a big variation in astigmatism was noted during the course, especially with cyclopentolate. This study suggests that 1% tropicamide should be a good agent for routine refractive status checking on myopic children.

Accommodation, Ocular↗

PubMed1998

The effect of intravitreal injection of atropine on the proliferation of scleral chondrocyte in vivo.

Atropine was found to be effective in arresting the progression of myopia. However, the actual mechanism is still unclear. Thus, we tried to investigate the in vivo effect of atropine on the proliferation of scleral chondrocytes in chicks of form-deprivation myopia. Twenty chicks were equally divided into 4 groups which included intravitreal injection of normal saline (IVN), IVN with goggling (IVNG), intravitreal injection of atropine (1%) (IVA), and IVA with goggling (IVAG) groups. Intravenous injection of bromodeoxyuridine (BrdU) (30 mg/kg) from subaxillary vein was performed 2 hours before being sacrificed. The eyeballs were then fixed in 10% neutral-buffered formalin at 4 degrees C. Standard BrdU immunohistochemical staining was performed. The BrdU labeling index was obtained from the average of positive labelings of BrdU in scleral chondrocytes for every 100 counting cells in posterior poles and anterior scleral margins by two experienced technicians. The BrdU index on the anterior scleral margin of the IVAG group was less than that of the IVNG group. The index on the anterior scleral margin of the IVNG group was higher than the IVN group. Although the index on the posterior poles of the IVNG group was also higher than the IVN group, it was statistically not different. Also, no statistical difference was found between IVN and IVA on the anterior scleral margins or posterior poles. The index was significantly different on the anterior scleral margins, but not on the posterior pole among each group. Therefore, intravitreal injection of atropine could inhibit the proliferation of chondrocytes on the anterior margins of sclera, but not the posterior poles in form-deprivation myopia.

Animals↗

PubMed1998

Influence of destruction of retina-RPE complex on the proliferation of scleral chondrocytes in chicks.

We studied the role of the retina-retinal pigment epithelium (RPE) complex in the proliferation of scleral chondrocytes in chicks. Seventy-two chicks were allocated to one of four groups: intravitreal gentamicin (400 microg) injection (destruction of retina-RPE complex); intravitreal gentamicin injection with goggling; goggling only (form-deprivation myopia); and intravitreal saline injection (control). The chicks were killed and retina-RPE complexes were harvested under a microscope. Retina-RPE complexes were then co-cultured with primary culture of first day scleral chondrocytes in Transwell-COL co-culture systems (Costar), with two different pore sizes (0.4 and 3.0 microm) and serum-deprivation medium. An MTT assay was performed at A550 after 4 days. In the 0.4 microm pore size system, the absorbency at A550 showed no differences between groups. However, in the 3.0 microm pore size system, the absorbency at A550s in the intravitreal gentamicin groups was significantly lower than in the control and the goggle groups (p<0.05), indicating that destruction of the retina-RPE complex inhibited chondrocyte proliferation. The absorbency in the goggle group was higher than in the control group (p<0.05). These results indicate that the retina-RPE complex exerts a positive effect on the proliferation of scleral chondrocytes via a molecule sized between 0.4.and 3.0 microm in diameter.

Animals↗

PubMed1998

Effects of lid suturing and trans-scleral cryotherapy on ocular growth in a piglet model.

PURPOSE: To study whether lid suturing can induce axial myopia and explore whether trans-scleral cryotherapy can affect ocular growth in piglets or not. METHODS: A total of fourteen 2- to 3-week-old piglets were studied. Three groups were included: lid suture, cryotherapy, and both lid suture/cryotherapy groups. The lid suturing group (N = 6) was given lid suturing to produce visual deprivation. The cryotherapy group (N = 4) received trans-scleral cryotherapy 360 degrees to encircle anterior to the equator. Both treatment groups (N = 4) received both cryotherapy and lid suturing. The cycloplegic refraction, corneal power, biometric axial length, and intraocular pressure (IOP) were measured before the experiments and 4 months later. RESULTS: Mild axial myopia was induced in five lid-sutured eyes of the lid suturing group. The ocular refraction and eye size of cryotherapy eyes were not different from the control eyes in the cryotherapy group. More myopia was found in all four piglets that received both cryotherapy and lid suturing. CONCLUSIONS: Lid suturing can induce axial myopia; however, cryotherapy did not affect normal eye growth and did not prevent the development of lid suturing myopia in piglets.

Animals↗

PubMed1998

Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs.

This article describes the nucleotide sequence of a porcine circovirus (PCV) which possesses a high degree of association with postweaning multisystemic wasting syndrome (PMWS), a newly described disease of young pigs. The DNA sequence of this PMWS-associated PCV (pmws PCV) has 68% homology with that of a previously published nonpathogenic strain of PCV. The strains appear to be closely related yet distinct from one another.

Amino Acid Sequence↗

PubMed1998

Production and properties of a raw-starch-degrading amylase from the thermophilic and alkaliphilic Bacillus sp. TS-23.

The optimum temperature and initial medium pH for amylase production by Bacillus sp. TS-23 were 55 degrees C and 8.5 respectively. Maximum amylase activity was obtained in a medium containing peptone and soluble starch as nitrogen and carbon sources. Activity staining revealed that two amylases with molecular masses of 150 and 42 kDa were produced when maltose, soluble starch or amylose was used as carbon source for growth, whereas only the 150 kDa protein was detected in the medium containing water-insoluble carbon sources. A raw-starch-degrading amylase was purified from culture supernatant of Bacillus sp. TS-23. The molecular mass of the purified amylase was estimated at 42 kDa by electrophoresis. The enzyme had a pI of 4. 2. The optimal pH and temperature for activity were 9.0 and 70 degrees C respectively. The thermoactivity of the purified enzyme was enhanced in the presence of 5 mM Ca2+; under this condition, enzyme activity could be measured at a temperature of 90 degrees C. The enzyme was strongly inhibited by Hg2+, Pb2+, Zn2+, Cu2+ and EDTA, but less affected by Ni2+ and Cd2+. The enzyme preferentially hydrolysed high-molecular-mass substrates with an alpha-1, 4-glucosidic bond except glycogen. The raw starches were partly degraded by the purified amylase to yield predominantly oligosaccharides with degrees of polymerization 3, 4 and 5.

Bacillus↗

PubMed1997

MADD, a novel death domain protein that interacts with the type 1 tumor necrosis factor receptor and activates mitogen-activated protein kinase.

The death domain of the type 1 tumor necrosis factor receptor (TNFR1) mediates interactions with several proteins involved in signaling the downstream effects of TNF. We have used the yeast interaction trap to isolate a protein, MADD, that associates with the death domain of TNFR1 through its own C-terminal death domain. MADD interacts with TNFR1 residues that are critical for signal generation and coimmunoprecipitates with TNFR1, implicating MADD as a component of the TNFR1 signaling complex. Importantly, we have found that overexpression of MADD activates the mitogen-activated protein (MAP) kinase extracellular signal-regulated kinase (ERK), and expression of the MADD death domain stimulates both the ERK and c-JUN N-terminal kinase MAP kinases and induces the phosphorylation of cytosolic phospholipase A2. These data indicate that MADD links TNFR1 with MAP kinase activation and arachidonic acid release and provide further insight into the mechanisms by which TNF exerts its pleiotropic effects.

Amino Acid Sequence↗