Biomedical subjects

S R Lin

Publications and source records attributed to S R Lin.

At least 37 records · Page 2Linked to original sources

PubMed2000

Up-regulation of adrenal cortical and medullary atrial natriuretic peptide and gene expression in rats with deoxycorticosterone acetate-salt treatment.

Our previous study demonstrated that human adrenal medulla is a site of atrial natriuretic peptide (ANP) synthesis. To further evaluate the role of adrenal ANP in body fluid homeostasis, we investigated the changes in adrenal ANP in rats receiving deoxycorticosterone acetate (DOCA)-salt treatment. In situ hybridization and immunohistochemical study showed that adrenal ANP messenger RNA (mRNA) and ANP-like immunoreactivities (ANP-LI) were mainly localized in the zona glomerulosa and medulla of vehicle-treated rats. DOCA-salt treatment activated ANP mRNA and peptide expression in all adrenal zones, especially in the zona fasciculata/reticularis from 12 h to the entire 8-day study period. Using a semiquantitative RT-PCR technique, the relative quantities of ANP mRNA in the adrenals of the DOCA-salt-treated group were significantly increased from 1 to 8 days, whereas the adrenal weights of DOCA-salt-treated rats were significantly decreased from day 2 to day 8. Our results are the first to indicate that ANP is synthesized not only in the adrenal medulla but also in the adrenal cortex and their syntheses are markedly increased in DOCA-salt-treated rats. These results imply that adrenal ANP may participate in the intraadrenal regulation of adrenal function on water-electrolyte homeostasis in an autocrine or paracrine manner.

Adrenal Cortex↗

PubMed2000

Mice deficient in hepsin, a serine protease, exhibit normal embryogenesis and unchanged hepatocyte regeneration ability.

Hepsin, a liver-enriched novel serine protease, has been implicated in participating with normal cell growth, embryogenesis, and blood coagulation pathway. To study its function in vivo, we have disrupted the mouse hepsin gene by homologous recombination. Targeted disruption of the hepsin gene and ablation of hepsin message were demonstrated by Southern blotting, Northern blotting and RT-PCR analysis. Homozygous hepsin -/- mice were viable, fertile, and exhibited no gross abnormalities, as judged by the size, weight and blood coagulation (PT) assays. However, the serum concentration of the bone form of alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase of the hepsin -/- mice was mildly elevated, in spite of no obvious pathological change of hepatocytes. To examine whether hepsin is involved in controlling cell growth in adult tissues, 70% hepatectomy was applied to the hepsin -/- mice. Liver regeneration proceeded normally in the hepsin -/- mice as judged by the liver mass restoration rate. These results suggest that loss of hepsin function causes no effect in cell growth and embryogenesis in vivo, which is in contradiction to the studies using in vitro cell culturing system. Moreover, gross mass regeneration of liver after damage proceeds normally in the absence of functional hepsin.

Animals↗

PubMed1999

Disulfide isomers of alpha-neurotoxins from King cobra (Ophiophagus hannah) venom.

Two novel alpha-neurotoxins, Oh-6A and Oh-6B, isolated from the king cobra (Ophiophagus hannah) venom, consist of 70 amino acid residues with 10 cysteine residues and share the same amino acid sequences as determined by Edman degradation on the peptide fragments generated from the proteolytic hydrolysates. Their sequences share 46-53% homology with Oh-4, Oh-5, Toxin a, and Toxin b from the same venom. The finding that Oh-6A and Oh-6B had different retention times in the reversed-phase column suggested that the two toxin molecules should not have the same conformation. Selective reduction on the disulfide bond, Cys26--Cys30, at the tip of their loop II structures resulted in the production of the partially reduced derivatives eluted at the same position. Under redox conditions, the partially reduced Oh-6A and 6B exclusively converted into native Oh-6A as evidenced by HPLC analyses. This suggests that Oh-6A and Oh-6B are disulfide isomers which probably arise from cis-trans isomerization of the Cys26--Cys30 disulfide bond. Alternatively, the two toxins exhibited binding activity toward nAChR and lethal toxicity equally. It reflects that the diversity around the extra loop at the loop II structure does not exert a significant effect on the manifestation of the neurotoxicity of Oh-6A and Oh-6B.

Amino Acid Sequence↗

PubMed1999

Determination of urinary arsenic, mercury, and selenium in steel production workers.

The fumes and dust of trace elements and their compounds are very toxic and have been related to an increase in the incidence of diseases. Occupational exposure to toxic metals and metalloids can be determined by means of workplace air measurements and biological monitoring. The aim of our investigation was to determine the concentrations of As, Hg, and Se in urine samples under routine clinical laboratory conditions. To assess the reliability of these methods, critical factors such as detection limit(s), calibration range(s), cost, accuracy, and precision were studied. The method was employed for the quantitative determination of arsenic, mercury, and selenium in urine samples from steel production and quality control workers and healthy unexposed controls. After pretreatment with acids, the samples were digested by means of a microwave oven. Arsenic was determined by hydride atomic absorption spectrometry and mercury was determined by cold vapor atomic absorption spectrometry, whereas selenium was determined by a graphite furnace atomic absorption spectrometry. The results indicate those urinary arsenic, mercury, and selenium levels of the exposed workers are significantly higher than those of the controls. The possibility that these elements are involved in the etiology of diseases is discussed and recommendations are made to improve workplace ventilation and industrial hygiene practices.

Arsenic↗

PubMed1999

The influence of age at onset and gender on the HLA-DQA1, DQB1 association in Chinese children with insulin dependent diabetes mellitus.

Certain alleles of human leukocyte antigen (HLA)-DR and -DQ genes have been strongly associated with susceptibility and resistance to insulin- dependent diabetes mellitus (IDDM). To further clarify the association of HLA DQ alleles with IDDM and the influence of age at onset and gender on the association with IDDM, we investigated the association of HLA-DQA1, -DQB1 in 54 childhood onset Chinese (21 male) IDDM patients and 65 normal controls by using polymerase chain reaction-sequence specific primer (PCR-SSP). The mean age plus or minus SD at onset of IDDM patients was 8.37+/-3.54 year old. Our results revealed that the frequencies of DQA1 *0301, *0302, DQB1 *0201, and *0302 in IDDM patients were significantly higher than that in the control group (p < 0.025, < 0.005, < 0.001, and < 0.001, respectively). The frequency of DQA1 *0301, *0302, DQB1 *0201, and *0302 were susceptible alleles to IDDM with relative risks of 2.0, 3.5, 5.0 and 4.3, respectively. The protective alleles to IDDM were DQA1 *0101, *0103, DQB1 *0301, *0503, and *0602. We divided IDDM patients into three groups according to age at onset (1-5, 6-10, and 11-15 years old). The frequency of DQA1 *0302 decreased as age increased, and the frequency of DQA1 *0501 increased as age increased. Our results also showed that male IDDM patients had higher frequencies of DQA *0501, DQB1 *0201 than female IDDM patients (p < 0.025 and < 0.025, respectively), while female IDDM patients had higher frequencies of DQB1 *0502 than male IDDM patients (p < 0.05). In our study significant susceptibility haplotypes to IDDM were DQA1 *0301-DQB1 *0302, DQA1 *0501-DQB1 *0201, DQA1 *0301-DQB1 *0201, and DQA *0302-DQB1 *0201.

Adolescent↗

PubMed1999

Chromosome 11q13 and atopic asthma.

Asthma is a complex syndrome in which bronchial inflammation and smooth muscle hyperactivity lead to labile airflow obstruction. The commonest form of asthma is that due to atopy, which is an immune disorder where production of IgE to inhaled antigens leads to bronchial mucosal inflammation. The ultimate origins of asthma are interactive environmental and genetic factors. The genetics is acknowledged to be heterogeneous, and one chromosomal region of interest and controversy has been 11q13. To clarify the nature of the chromosome 11q13 effect in atopy and asthma, we conducted a genetic association study in subjects with marked atopic asthma and matched controls, which incorporated the study of 13 genetic variants over a distance of 10-12 cM and which took account of detailed immune and clinical phenotyping. Association with high IgE levels was limited to the interval flanked by D11S1335 and CD20 in a 0.8-Mb interval and was greatest for variants of Fc epsilonRIbeta and HTm4; these variants also associated with asthma (recurrent wheeze with labile airflow obstruction and need for regular inhaler treatment). At the more telomeric marker, D11S480, variants associated with asthma, but not with high IgE levels. The data might support the possibility of multiple loci relevant to atopic asthma on chromosome 11q13.

Alleles↗

PubMed1999

Mutational spectrum of p53 gene in arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan.

To understand the role of p53 tumour suppressor gene in the carcinogenesis of arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan, we collected tumour samples from 23 patients with Bowen's disease, seven patients with basal cell carcinomas (BCC) and nine patients with squamous cell carcinomas (SCC). The result showed that p53 gene mutations were found in 39% of cases with Bowen's disease (9/23), 28.6% of cases with BCC (2/7) and 55.6% of cases with SCC (5/9). Most of the mutation sites were located on exon 5 and exon 8. Moreover, the results from direct sequencing indicated that missense mutations were found at codon 149 (C-->T) in one case, codon 175 (G-->A) in three cases, codon 273 (G-->C) in three cases, codon 292 (T-->A) in one case, codon 283 (G-->T) in one case, codon 172 (T-->C) in one case and codon 284 (C-->A) in one case. In addition, silent mutations were also found in four cases. These mutations were located at codons 174, 253, 289 and 298 respectively. In immunohistochemistry analysis, p53 overexpression was found in 43.5% (10/23) of cases with Bowen's disease, 14% (1/7) of cases with BCC and 44% (4/9) of cases with SSC. These findings showed that p53 gene mutation rate in arsenic-related skin cancers from the blackfoot disease endemic area of Taiwan is high and that the mutation types are different from those in UV-induced skin cancers.

Aged↗

PubMed1999

Increased nitric oxide synthase mRNA expression in the renal medulla of water-deprived rats.

UNLABELLED: Increased nitric oxide synthase mRNA expression in the renal medulla of water-deprived rats. BACKGROUND: Experiments were performed to investigate whether renal nitric oxide synthase (NOS) mRNA and protein expression are responsive to the alteration of body volume. METHODS: Four days of water deprivation (WD) was initiated in 16 male Wistar rats, and 16 normal rats (NC) served as the control group. Neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS) mRNAs and immunoreactivity were measured by reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern blot hybridization and immunohistochemistry, respectively. Plasma angiotensin II, vasopressin, and atrial natriuretic peptide (ANP) concentrations were measured by radioimmunoassay. RESULTS: The four-day WD increased plasma sodium and osmolality levels, but severely decreased daily urine sodium excretion and urine volume. Plasma angiotensin II and vasopressin concentrations were increased, but the plasma ANP level was significantly decreased in WD rats. nNOS, eNOS, and iNOS mRNA levels were increased by 5.2-, 3.3-, and 3. 4-fold in the outer medulla and 1.7-, 1.5-, and 1.8-fold in the inner medulla, whereas no significant difference was found in the renal cortex of WD rats as compared with NC rats. Additionally, immunohistochemistry revealed that the immunostaining intensity of nNOS, eNOS, and iNOS was clearly enhanced in the medullary thick ascending limb, proximal straight tubule, inner medullary collecting duct, and proximal convoluted tubule in WD rats. Kidney angiotensin II content as well as renin mRNA levels in renal cortex, outer medulla, and inner medulla in WD rats were apparently increased. CONCLUSIONS: Our results indicate that the increases of nNOS, eNOS, and iNOS synthesis in the kidney, particularly in the renal medulla, may have a role in the adaptation of renal function to volume depletion in the face of an increase of systemic and intrarenal vasoconstrictive substances.

Angiotensin II↗

PubMed1999

Seroepidemiologic survey for hepatitis B virus infection in Taiwan: the effect of hepatitis B mass immunization.

A hepatitis B mass immunization program was launched in Taiwan in July 1984, beginning with newborns of hepatitis B carrier mothers for the first 2 years of the program, which was then extended to all newborns. Seroepidemiology was studied in 3 cohorts at age 6 years. Each cohort consisted of 1500 children proportionally and randomly sampled from those entering elementary school in 1989, 1991, and 1993, representing those born 1 year before the program began and years 1 and 3 of the program, respectively. By RIA, the hepatitis B surface antigen positivity rates in the groups were 10.5%, 6.3%, and 1.7%, respectively; hepatitis B surface antibody positivity rates were 36.9%, 62.0%, 65.4%; and hepatitis B infection rates were 25.0%, 15.9%, 4.3%. Thus, universal immunization was more effective in reducing hepatitis B carriage than selective immunization of newborns of carrier mothers only. The program has proved effective in controlling chronic hepatitis B infection in Taiwan.

Biomarkers↗

PubMed1999

The hemodynamic changes of gastroduodenal regional blood flow after Helicobacter pylori eradication in patients with duodenal ulcer scar.

Duodenal ulcer (DU) is frequently accompanied by Helicobacter pylori (Hp) infection and associated with the imbalance of aggressive factors and defensive factors. To investigate the possible relationship between Hp and regional gastric blood flow, 26 endoscopically proved DU (scar stage) and Hp infection patients were included and received triple therapy (colloid bismuth subcitrate 120 mg qid for 4 weeks, amoxicillin 500 mg qid for 2 weeks and metronidazole 250 mg tid for 2 weeks). Regional gastroduodenal blood flow (RGDF) was measured at DU scar area and antrum lesser curvature site by laser Doppler flowmetry during endoscopic examination, before and one month after triple therapy. In 22 patients with Hp eradication the RGDF was significantly elevated at antrum lesser site after triple therapy (p < 0.05) but there was no difference at DU scar area. However, in 4 patients without Hp eradication no difference of RGDF in these two points was found. Therefore, Hp appears to have direct effects on gastric microcirculation.

Adult↗

PubMed1999

Stability of self-prepared fortified antibiotic eyedrops.

Self-prepared fortified antibiotic eye drops are essential for the severe ocular infection. The relation of potency decay and storage conditions including temperature, concentration and duration were studied by the changes of MIC. 1% and 10% amikin, 10% and 50% pipril, and 5% and 25% vancomycin were diluted from the prarenteral antibiotics with the 5% glucose and storage at 4, -18 and -80 degrees C for 3, 7, 14 and 28 days. MICs of amikin against Pseudomonas aeruginosa, pipril and vancomycin against Staphylococcus aureus were determined by the agar diffusion method. Fluctuations of the MIC were noted during the observation period. Most of the significant changes of MIC were found during the first 7 days. When the potencies between time of zero and 28 days are compared, only 10% pipril and 25% vancomycin stored at -80 degrees C had significant change. Our conclusion is that all of the fortified antibiotic eye drops in this study can be stored in the house refrigerator or freezer for up to 28 days. High concentration may show a negative result of the preservation. Temperature does not influence the preservative effect within four weeks' observation.

Anti-Bacterial Agents↗

PubMed1999

Fungal keratitis caused by Candida utilis--case report.

We report the first case of an ocular fungal infection caused by Candida utilis in a 72-year-old woman. Although candidiasis is the most common opportunistic fungal infection in humans, C. utilis had not been associated with human morbidity until two cases of fungemia were reported in 1988 and 1993. The clinical features exhibited typical feather-like infiltration at the ulceration margin of this case. After treatment with topical fluconazole and amphotericin-B, the ulceration healed within 3 weeks. The patient underwent penetrating keratoplasty and regained useful vision. We concluded that C. utilis is a new corneal pathogen and should be included in the differential list of mycotic keratitis.

Aged↗

PubMed1998

Human ARHGDIG, a GDP-dissociation inhibitor for Rho proteins: genomic structure, sequence, expression analysis, and mapping to chromosome 16p13.3.

GDP-dissociation inhibitors (GDIs) play a primary role in modulating the activity of GTPases. We recently reported the identification of a new GDI for the Rho-related GTPases named RhoGDIgamma. This gene is now designated ARHGDIG by HUGO. Here, in a detailed analysis of tissue expression of ARHGDIG, we observe high levels in the entire brain, with regional variations. The mRNA is also present at high levels in kidney and pancreas and at moderate levels in spinal cord, stomach, and pituitary gland. In other tissues examined, the mRNA levels are very low (lung, trachea, small intestine, colon, placenta) or undetectable. RT-PCR analysis of total RNA isolated from exocrine pancreas and islets shows that the gene is expressed in both tissues. We also report the genomic structure of ARHGDIG. The gene spans over 4 kb and is organized into six exons and five introns. The upstream region lacks a canonical TATA box and contains several putative binding sites for ubiquitous and tissue-specific factors active in central nervous system development. Using FISH, we have mapped the gene to chromosome band 16p13.3. This band is rich in deletion mutants of genes involved in several human diseases, notably polycystic kidney disease, alpha-thalassemia, tuberous sclerosis, mental retardation, and cancer. The promoter structure and the chromosomal location of RhoGDIgamma suggest its importance and underscore the need for further investigation into its biology.

Base Sequence↗

PubMed1998

Unfolding/folding studies on cobrotoxin from Taiwan cobra venom: pH and GSH/GSSG govern disulfide isomerization at the C-terminus.

Refolding of cobrotoxin was assessed by the exposure degree of its single Trp determined by an acrylamide quenching study. The change in the accessibility of Trp for acrylamide quantitatively reflected the formation of folded cobrotoxin, and the data were confirmed by HPLC and gel electrophoresis analyses. However, the site-specific information provided by quenching Trp fluorescence revealed that the ordered structure in the neighborhood of Trp was attained prior to the complete formation of the tertiary structure of cobrotoxin. HPLC analyses showed that, in addition to refolded cobrotoxin, two novel species (cobrotoxin II and cobrotoxin III) with isomerization of disulfide bonds at the C-terminus of the toxin molecule were produced along the folding reaction. The disulfide pairings in cobrotoxin II and cobrotoxin III were Cys43-Cys55 and Cys54-Cys60 and Cys43-Cys60 and Cys54-Cys55, respectively. Among the three possible two-disulfide species at the C-terminus, the disulfide linkages Cys43-Cys60 and Cys54-Cys55 of cobrotoxin III caused a marked decrease in lethality and resulted in a conformation which was notably different from that observed with the native toxin molecule as evidenced by CD spectra. The refolding reaction was accelerated by the addition of GSH/GSSG, and the resulting products were mostly folded cobrotoxin. However, if GSH/GSSG was not added into the initial folding materials, the yields of cobrotoxin II and cobrotoxin III greatly increased. The conversion of cobrotoxin to its isomers was to be irreversible and pH-dependent: the higher the pH, the faster the rate of conversion. However, this conversion could be partly inhibited by GSH/GSSG. Cobrotoxin II and cobrotoxin III were purified from Taiwan cobra venom as well, and their yields in comparison to that of cobrotoxin in venom were similar to that noted with the folded products in the presence of GSH/GSSG. Moreover, the rate of disulfide isomerization was expected to be slow in venom fluid in which the pH was approximately pH 6.2. Thus, the finding that cobrotoxin represents the predominant neurotoxin species in Taiwan cobra venom is probably associated with the synergistic effects of GSH/GSSG and pH.

Amino Acid Sequence↗

PubMed1998

Mutations of K-ras oncogene in human adrenal tumours in Taiwan.

Recently, we have found a high frequency of p53 gene mutations in human functional adrenal tumours. As the tumorigenesis is a multigene defect, we believe that other oncogenes may also be involved in the initiation or progression of adrenal tumours. Using the single-strand conformational polymorphism (SSCP) method, we chose the ras oncogenes as the target in this screening procedure because their high mutation rates were detected in thyroid tumours. For the ras oncogenes analysed, exon 1 to exon 2 of H-ras and K-ras genes in the tumour tissues of 13 Conn's syndrome, two adrenal Cushing's syndrome, two non-functional adrenal tumours, one adrenocortical hyperplasia and eight phaeochromocytomas and its paired adjacent normal adrenal tissues were amplified and sequenced. No mutations were detected in the H-ras gene. But mutations of the K-ras gene were detected in 46% (6 of 13) of Conn's syndrome; the hot spots were located at codon 15, 16, 18 and 31, which were different from those previously found in other tumours (codon 12, 13 and 61). Northern blot analysis with 1.1 kb K-ras cDNA revealed that K-ras mRNA was more than tenfold over-expressed in four of Conn's syndrome, one case of Cushing's syndrome and one case of adrenocortical hyperplasia. The mutation sites and mutation type were not found in other tissues, which conferred that this was highly related to adrenocortical tumours. Yet, the correlation between K-ras oncogene and adrenocortical tumours needs to be clarified by further studies.

Adrenal Cortex Neoplasms↗

PubMed1998

Structural factors affect the interactions of anticardiotoxin antibodies and cobra venom cardiotoxins.

Two antibody preparations against cardiotoxins were raised by immunizing rabbits with cardiotoxin 1 and cardiotoxin 3, respectively. The two antibody preparations showed precipitin reactions with cardiotoxins 1, 2, 3 and 5, respectively. However, the results of competitive enzyme-linked immunoassay revealed that the respective cardiotoxin molecules exhibited different reactivity toward anticardiotoxin antibodies. Moreover, the order of reactivity with antibodies was not in line with the degree of their sequence identity. This suggest that the anticardiotoxin antibodies may recognize conformational epitopes rather than sequential ones in the toxin molecules. Alternatively, the four cardiotoxins reacted well with the antibodies in the absence of competitor, suggesting that sequence variations with cardiotoxin molecules may not exclusively influence the potential use of the anticardiotoxin antibodies for the neutralization of the activity of cardiotoxin variants.

Amino Acid Sequence↗

PubMed1998

Characterization and cloning of long neurotoxin homolog from Naja naja atra.

The cDNA encoding a long neurotoxin homolog was constructed from the cellular RNA isolated fom the venom glands of Naja naja atra (Taiwan cobra) by reverse transcription-polymerase chain reaction. BLAST searches for sequence similarity in the GenBank databases reveal that the cDNA sequence of the long neurotoxin homolog is not highly homologous with long and short neurotoxins. Although the long neurotoxin homolog exhibited an activity to inhibit acetylcholine-induced muscle contractions as Naja naja atra cobrotoxin, the degree of inhibition caused by the addition of long neurotoxin homolog was only approximately 35% of that observed with the addition of cobrotoxin. Moreover, the primary structure of the long neurotoxin homolog did not fulfill the characteristic features of long or short neurotoxins. Together with long neurotoxin homologs from other snake species, they probably represent an evolutionary divergence between long and short neurotoxins.

Acetylcholine↗

PubMed1998

Cytokines in murine lyme carditis: Th1 cytokine expression follows expression of proinflammatory cytokines in a susceptible mouse strain.

The cardiac infiltrate seen in murine Lyme carditis is composed predominantly of macrophages, but small numbers of T cells are also present. To identify the cytokines present in cardiac lesions from susceptible mice, semiquantitative polymerase chain reaction was done on cardiac tissue from mice infected with Borrelia burgdorferi. The temporal expression of proinflammatory and T cell-derived cytokines was characterized in cardiac tissue at days 0, 3, 7, 14, 21, and 42 after infection with B. burgdorferi. Early in the course of infection, up-regulation of the proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha was detected. The Th1 cytokine interferon-gamma appeared after the expression of the proinflammatory cytokines and remained elevated throughout the study. Interleukin-4 was not detectable at any time in cardiac lesions. These data are the first to identify cytokines expressed at the lesional level in murine Lyme carditis and to demonstrate a Th1 pattern of cytokine expression in this lesion.

Animals↗