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Biomedical subjects

Y C Liu

Publications and source records attributed to Y C Liu.

At least 19 recordsLinked to original sources

Comparison of different protein immobilization methods on quartz crystal microbalance surface in flow injection immunoassay.

In this study, a quartz crystal microbalance (QCM) system operated repetitively in flow injection analysis (FIA) mode, is reported. Four immobilization approaches of seven different methods include: (i) physical adsorption; (ii) two thioamine thiolation methods, using cysteamine and cystamine for gold chemisorption and further coupling; (iii) two oxidized dextran spacer methods, coupling of cysteamine and cystamine thiolated QCM surface with periodate-oxidized dextran for further Schiff acid-base reaction; and (iv) two thiol-gold chemisorption-based self-assembled monolayer (SAM), applying short-chain, C(3), and long-chain, C(11), mercapto fatty acids to insolubilize human serum albumin (HSA) on QCM surface. Effects of these protein immobilization methods on FIA immunoassay of anti-HSA were compared. At the 0.01 mg/ml anti-HSA level, the lowest analyte concentration tested, the SAM using 11-mercaptoundecanoic acid as QCM surface activating agent generated a larger frequency shift than the other immobilization methods. This implied that the use of thiolated long-chain fatty acid constructed as self-assembled monolayer may thereby potentially be a useful protein immobilization method in QCM-FIA application.

Absorption↗

Three-dimensional mapping of brain neuropils in the cockroach, Diploptera punctata.

Herein, we present a complete three-dimensional (3D) map of major neuropil structures in the central brain of the cockroach Diploptera punctata. The positions of the structures have been ascertained by confocal microscopy, which, until now-for reasons of tissue opacity and nonhomogeneity-has been thought impractical in imaging fluorescently labeled structures thicker than 150 microm. In this report, however, we have used digestive enzymes and microwave-aided fixation to stain, clear, and optically section, in its entirety, an intact central brain more than 500 microm thick. The central brain from an adult female cockroach was stained thoroughly with the membrane probe NBD-ceramide and the DNA probe propidium iodide. The central brain as well as such neuropil regions as mushroom bodies, central complex, antennal glomeruli, and lobus glomerulati were individually outlined, segmented, and reconstructed in three dimensions to a spatial resolution of approximately 1 microm in the X-Y plane and 3 microm in the Z plane. The volume and surface area of each neuropil compartment were determined, and Kenyon cells of the mushroom bodies were counted. We determined that each brain hemisphere contains about 230,000 Kenyon cells, 99 antennal lobe glomeruli, and 40 lobus glomerulatus glomeruli. Segmented compartments were assigned as separate channels and merged into a single data base to reconstruct a 3D central brain containing eight different channels. This is the first 3D map at submicron resolution of an entire animal's brain that measures more than 500 microm in thickness.

Animals↗

Inappropriate use of antibiotics and the risk for delayed admission and masked diagnosis of infectious diseases: a lesson from Taiwan.

BACKGROUND: Antibiotic resistance is a serious problem worldwide. It is particularly alarming in Taiwan and other countries of the Pacific Rim, where antimicrobial drugs are used excessively. OBJECTIVE: To determine whether use of antimicrobial drugs before coming to an emergency department was associated with delayed admission or masked or missed diagnoses at a large general hospital in Taiwan. METHODS: Antimicrobial activity in urine (AAU) was determined in all patients seen in the emergency department during a 3-month study. A physician, unaware of the results of the urine tests, reviewed the medical charts of patients who were admitted to the hospital to determine whether admission was delayed for at least 7 days or the diagnosis was masked or missed. RESULTS: Of the 1182 patients, 444 were admitted to the hospital. In 220 patients (49.5%), AAU was detected. There was no significant difference in AAU between patients with or without an infectious disease (53.0% vs 46.3%, respectively; P =.41). For patients with infection, 34.8% of those with AAU had a delayed admission, compared with only 21.6% without AAU (relative risk [RR], 1.61; 95% confidence interval [CI], 1.03-2.52; P =.03). For patients without infection, 36.2% of those with AAU had a delayed admission compared with 31.1% without AAU (RR, 1.16; 95% CI, 0.81-1.68; P =.64). For patients with infection, 48.7% of those with AAU had a masked or missed diagnosis, compared with 25.5% without AAU (RR, 1.91; 95% CI, 1.30-2.80; P<.001). For patients without infection, 27.6% of those with AAU had a masked or missed diagnosis compared with 14.8% without AAU (RR, 1.87; 95% CI, 1.11-3.17; P =.02). CONCLUSION: Use of antimicrobial drugs before coming to an emergency department was associated with a significantly increased risk for delayed and masked or missed diagnoses of infectious diseases and missed diagnosis of noninfectious diseases.

Adolescent↗

Detection of elevated serum beta-chemokine levels in seronegative Chinese individuals exposed to human immunodeficiency virus type 1.

The mutations in the CCR5 coding region, such as CCR5Delta32 and CCR5m303, that suppress the transmission of human immunodeficiency virus (HIV) type 1 do not exist in Chinese people. However, 9 Chinese subjects in Taiwan with histories of multiple sexual exposures to HIV remained uninfected, suggesting that certain anti-HIV factors do indeed exist. Experiments were therefore designed to investigate the immune mechanism that protects this cohort against HIV infection. Peripheral blood samples from these 9 subjects and 7 healthy people who had not been exposed to HIV were obtained for the quantitation of the levels for beta-chemokines and interleukin 16 (IL-16) in serum samples or secreted by peripheral blood lymphocytes. Significantly higher serum levels for nearly all 3 beta-chemokines, regulation on activation, normal T cell-expressed and secreted, macrophage inflammatory protein (MIP)-1alpha, and MIP-1beta (P<.05, P<.05, and P=.05, respectively), but not IL-16, were detected in the 9 HIV-uninfected subjects as compared with control subjects. The result suggests that among the host genes and cellular factors thus far identified, beta-chemokines are the major HIV-suppressive factors that protect Chinese people from infection with HIV.

Adult↗

Cbl promotes ubiquitination of the T cell receptor zeta through an adaptor function of Zap-70.

Triggering of the T cell antigen receptor (TCR).CD3 complex induces its ubiquitination. However, the molecular events that lead to ubiquitin conjugation to these cell surface molecules have not been defined. Here we report that Cbl, a RING-type E3 ubiquitin-protein ligase, promotes ubiquitination of TCR zeta chain, which requires its functional variant Src homology 2 domain and an intact RING finger. The tyrosine kinase Zap-70, which binds to both TCR zeta and Cbl, plays an adaptor role in these events. Mutations in TCR zeta, Zap-70, or Cbl that disrupt the interaction between TCR zeta and Zap-70 or between Zap-70 and Cbl reduce ubiquitination of TCR zeta. Our results suggest a novel mechanism by which Cbl negatively regulates T cell development and activation by inducing ubiquitination of the TCR.CD3 components.

Cell Line↗

On-line concentration of s-triazine herbicides in micellar electrokinetic chromatography using a cationic surfactant.

On-line concentration of neutral species of s-triazine herbicides in micellar electrokinetic chromatography using tetradecylammonium bromide (TTAB) as a cationic surfactant was investigated. Factors affecting the stacking of analytes were examined. The results indicate that the stacking efficiency is markedly improved with addition of phosphate buffer in the sample matrix. It was found that, depending on the nature of the analytes, the most effective stacking of these analytes occurs when the ratio of the conductivity of buffer electrolyte to that of sample matrix is in the range 1.4-1.2, with sample matrix containing phosphate buffer. Micelle concentration in the separation buffer is also a crucial factor to enhance the stacking efficiency and detection sensitivity of analytes. Moreover, the stacking efficiency of each individual analyte depends on its binding constant to TTAB micelles. The concentration effect is primarily based on sweeping mechanism which is operated in a normal stacking mode with reversed electrode polarity in the presence of reversed electroosmotic flow. As a result of concentration enhancement, the detection limits of these herbicides can reach about 9-15 ng/ml with UV detection.

Chromatography, Micellar Electrokinetic Capillary↗

Comparative study of the effects of auditory, visual and multimodality displays on drivers' performance in advanced traveller information systems.

A simulator study was conducted to compare 16 younger (mean age 22 years) and 16 older (mean age 68 years) drivers' ratings of workload (time, visual, psychological stress) and performance of navigation and button-pushing (identification of vehicle or road hazards) tasks under both high- and low-load driving conditions when simple or complex advanced traveller information (ATI) was presented visually only, aurally only or by multimodality (visual and auditory) display. For all participants, both the auditory and multimodality displays produced better performance in terms of response times, total number of correct turns and subjective workload ratings than those of using the visual-only display. Participants using the multimodality display also made the fewest errors related to push-button and navigation tasks, and controlled their vehicles properly. The visual display led to less safe driving, apparently because it imposed higher demands on the drivers' attention. An age effect was found in the present study, with younger drivers performing better and reporting less stress than older drivers. Notably, however, use of the multimodality display significantly improved the older drivers' performance in the button-pushing task.

Adult↗

Temporal trends and molecular epidemiology of HIV-1 infection in Taiwan from 1988 to 1998.

Eight hundred and seventy-nine HIV-1-infected patients (comprising 46% of reported HIV-1/AIDS cases in Taiwan) were recruited for this study of the molecular epidemiology of HIV-1 in Taiwan from 1988 to 1998. HIV-1 subtypes were determined using a modified peptide-enzyme immunoassay complemented with DNA sequencing and phylogenetic analysis. Of the 807 HIV-1 infected men, 68.2% were infected with HIV-1B, 29.5% with HIV-1 circulating recombinant form (CRF)01_AE and 2.3% with other subtypes. Of the 72 HIV-1-infected women, 72.2% were infected with HIV-1 CRF01_AE, 13.9% with HIV-1B, and 13.9% with other subtypes. All of 8 foreign-born, Southeast Asian women and 6 of 7 (85.7%) Taiwan-native female commercial sex workers were infected with HIV-1 CRF01_AE. Fourteen of the 33 (42.4%) heterosexual married men with CRF01_AE had transmitted HIV-1 to their wives, whereas only 1 of 17 (5.9%) men with HIV-1 B had transmitted HIV-1 to their spouses (p < .01). Of 18 heterosexual male injecting drug users, 1 of 12 (8.5%) with HIV-1B and 5 of 6 (83.3%) with HIV-1 CRF01_AE had had sexual contact with female commercial sex workers (p < .01). Therefore, in this population, CRF01_AE was preferentially associated with heterosexual risk groups, a finding compatible with differences in transmission capability between B and non-B subtypes.

Adult↗

Optimization of separation and migration behavior of chloropyridines in micellar electrokinetic chromatography.

The separation and migration behavior of pyridine and eight chloropyridines, including three monochloropyridines, four dichloropyridines, and 2,3,5-trichloropyridine were investigated by micellar electrokinetic chromatography using either sodium dodecyl sulfate (SDS) as an anionic surfactant or SDS-Brij 35 mixed micelles. Various parameters such as buffer pH, SDS concentration, Brij 35 concentration and methanol content that affect the separation were optimized. Complete separation of these chloropyridines was optimally achieved with a phosphate buffer containing SDS (30 mM) and methanol (10%, v/v) at pH 7.0. The resolution and selectivity of analytes could be considerably affected by the addition of methanol and/or Brij 35 to the background electrolyte. The migration order of these chloropyridines depends primarily on their hydrophobicity. However, electrostatic interactions may also play a significant role in the determination of the migration order of the positional isomers of chloropyridines.

Buffers↗

Regulation of Cbl molecular interactions by the co-receptor molecule CD43 in human T cells.

CD43, one of the most abundant glycoproteins on the T cell surface, has been implicated in selection and maturation of thymocytes and migration, adhesion, and activation of mature T cells. The adapter molecule Cbl has been shown to be a negative regulator of Ras. Furthermore, it may also regulate intracellular signaling through the formation of several multi-molecular complexes. Here we investigated the role of Cbl in the CD43-mediated signaling pathway in human T cells. Unlike T cell receptor signaling, the interaction of the adapter protein Cbl with Vav and phosphatidylinositol 3-kinase, resulting from CD43-specific signals, is independent of Cbl tyrosine phosphorylation, suggesting an alternative mechanism of interaction. CD43 signals induced a Cbl serine phosphorylation-dependent interaction with the tau-isoform of 14-3-3. protein. Protein kinase C-mediated Cbl serine phosphorylation was required for this interaction, because the PKC inhibitor RO-31-8220 prevented it, as well as 14-3-3 dimerization. Moreover, mutation of Cbl serine residues 619, 623, 639, and 642 abolished the interaction between Cbl and 14-3-3. Overexpression of Cbl in Jurkat cells inhibited the CD43-dependent activation of the mitogen-activated protein kinase (MAPK) pathway and AP-1 transcriptional activity, confirming nevertheless a negative role for Cbl in T cell signaling. However, under normal conditions, PKC activation resulting from CD43 engagement was required to activate the MAPK pathway, suggesting that phosphorylation of Cbl on serine residues by PKC and its association with 14-3-3 molecules may play a role in preventing the Cbl inhibitory effect on the Ras-MAPK pathway. These data suggest that by inducing its phosphorylation on serine residues, CD43-mediated signals may regulate the molecular associations and functions of the Cbl adapter protein.

14-3-3 Proteins↗

Endothelial exposure to hypoxia induces Egr-1 expression involving PKCalpha-mediated Ras/Raf-1/ERK1/2 pathway.

Hypoxia induces endothelial dysfunction that results in a series of cardiovascular injuries. Early growth response-1 (Egr-1) has been indicated as a common theme in vascular injury. Here we demonstrates that in bovine aortic endothelial cells (ECs) subjected to hypoxia (PO(2) approximately 23 mmHg), rapidly increased Egr-1 mRNA expression which peaked within 30 min and decreased afterwards. Treatment of ECs with PD98059, a specific inhibitor to mitogen-activated protein kinase (MAPK/ERK), inhibited this hypoxia-induced Egr-1 expression. The involvement of ERK pathway was further substantiated by the inhibition of Egr-1 promoter activities when ECs were co-transfected with a dominant negative mutant of Ras (RasN17), Raf-1 (Raf 301), or a catalytically inactive mutant of ERK2 (mERK). In addition, the hypoxia-induced transcriptional activity of Elk-1, an ERK substrate, was abolished by administration of PD98059. Addition of calphostin C, a protein kinase C (PKC) inhibitor, completely blocked the hypoxia-augmented Egr-1 expression. The likewise occurred while exposing ECs to D609 to inhibit phospholipase C and BAPTA/AM to chelate intracellular calcium. Hypoxia to ECs increased ERK phosphorylation within 10 min and which was abolished by administration of PD98095, calphostin C, and BAPTA/AM. Hypoxia triggered a transient translocation of PKCalpha from cytosol to membrane fraction concurrent with the association of PKCalpha to Raf-1. Involvement of PKCalpha in mediating ERK activation was further confirmed by the inhibition of ERK and the subsequent Egr-1 gene induction with antisense oligonucleotides to PKCalpha. These results indicate that ECs under hypoxia induce Egr-1 expression and this induction requires calcium, phospholipase C activation, and PKCalpha-mediated Ras/Raf-1/ERK1/2 signaling pathway. Our finding support the importance of specific PKC isozyme linked to MAPK pathway in the regulation of endothelial responses to hypoxia.

Animals↗

Stereoselective reduction with NADH model BNAH through chiral induction in cyclodextrins.

Stereoselective reductive debromination-cyclopropanation of 2-bromo-1-phenylethylidene-malononitrile and 2-bromo-1-beta-naphthylethylidenemalononitrile by coenzyme NADH model BNAH through chiral induction in cyclodextrins is reported. The matching between substrates and cyclodextrins, the substituent effect, and the effect of cyclodextrin concentration on the optical yields have been investigated.

Cyclodextrins↗

In situ detection of hTERT mRNA relates to Ki-67 labeling index in papillary thyroid carcinoma.

BACKGROUND: Telomerase is activated in most human cancers but is inactivate in adult somatic tissues except for some proliferating cell lineages. The maintenance of telomerase activity may be a critical step of cellular immortalization and transformation. MATERIALS AND METHODS: We analyzed the expression of human telomerase reverse transcriptase (hTERT) using in situ hybridization and compared it to Ki-67 immunoreactivity in 29 cases of papillary thyroid carcinoma (PTC) and 17 cases of benign thyroid disease. RESULTS: The hTERT messenger RNA (mRNA) was expressed in the cytoplasm of carcinoma cells with moderate (n = 10) to strong intensity (n = 10) in 69% (20 of 29) PTC cases. Human TERT was found in only 29% (5 of 17) cases of benign thyroid disease. Human TERT gene expression was preferentially detected in PTC (P = 0.021). The Ki-67 labeling index was observed in 16 cases of PTC (16 of 29; 55.2%). This result was significantly different from that of benign thyroid disease (P = 0.014). The Ki-67 labeling index related to the intensity of hTERT mRNA expression (r = 0.51; P = 0.005) and was inversely associated with the follicular variant of PTC (r = -0.413; P = 0.026). No statistically significant difference was found between hTERT expression and histological subtype of PTC. CONCLUSIONS: Our results demonstrated that expression of hTERT could be detected using in situ hybridization in PTCs and was significantly distinguishable from that of benign thyroid disease. Human TERT expression was related to the Ki-67 labeling index, indicating that coupling of telomerase activation with cell proliferation was the associated mechanism for tumorigenesis.

Adolescent↗

An efficient method for the culture of Helicobacter pylori from gastric biopsies with two-section petri dishes.

PURPOSE: Helicobacter pylori is the principal cause of chronic gastritis and is strongly associated with peptic ulcer disease and gastric cancer, as well as with gastric lymphoma (mucosa-associated lymphoid tissue [MALT] type). H. pylori could be recovered by many kinds of media, however, it could not be detected efficiently. This study was trying to ameliorate the isolation and detection rate with selective and non-selective media. METHODS: First, type strain (ATCC 43504) was cultured with twelve different kinds of sheep blood agar based on brain heart infusion medium for five days. Then five better non-selective media and five selective media were chosen for the biopsy, which were incubated at 37 degrees C in a microaerophilic incubator for five days. Endoscopic biopsies were collected from 623 patients at Kaohsiung Medical University Hospital. RESULTS: It was found that non-selective media, which contain 5% modified chocolate agar (the ratio of fresh and heated sheep blood is 2) with 1% Isovitalex, had the best detection rate (91.1 +/- 0.6)%. Furthermore, 5% modified chocolate agar with 1% Isovitalex and 1% antibiotics would increase the detection rate to (97.1 +/- 0.3)%. CONCLUSIONS: According to Ansorg's method, the best detection rate (98.4 +/- 0.7)% could be obtained with two-section petri dishes which had 5% modified chocolate agar with antibiotics and the other without antibiotics.

Animals↗

Eosinophilic meningitis caused by Angiostrongylus cantonensis: report of 17 cases.

PURPOSE: To describe two outbreaks of Angiostrongylus cantonensis infection that occurred in Kaohsiung, Taiwan, during 1998 and 1999, and to characterize the source of the outbreaks and the clinical manifestations of the disease. SUBJECTS AND METHODS: We performed a retrospective cohort study among Thai laborers with eosinophilic meningitis who ate raw snails (Ampullarium canaliculatus), as well as an environmental surveillance of larvae in snails. RESULTS: We enrolled 17 Thai laborers in whom severe headache and eosinophilia developed within 4 to 23 days after eating raw snails. Twelve (71%) developed eosinophilic meningitis. Third-stage larvae were found in the cerebrospinal fluids of 2 patients and in all 12 tested snails. Specific antibodies to A. cantonensis were detected in serum from 16 of the patients and in cerebrospinal fluid from 5 of the patients. Central nervous system manifestations included headache (n = 17 [100%]), fever (n = 11 [65%]), Brudzinski's sign/stiff neck (n = 11 [65%]), hyperesthesia (n = 3 [18%]), cranial nerve palsy (n = 2 [12%]), diplopia (n = 2 [12%]), and ataxia (n = 1 [6%]). Laboratory findings included peripheral eosinophilia (n = 15 [88%]) and cerebrospinal fluid eosinophilia (n = 12 [71%]); elevated immunoglobulin (Ig) E levels (n = 13 [100%]); and transient increases in white blood cell count (n = 7 [41%]) and in serum levels of creatine kinase (n = 7 [41%]), transaminase (n = 3 [18%]), and lactate dehydrogenase (n = 2 [12%]). The severity of illness and eosinophilia were correlated with the number of ingested snails. Meningeal and basal ganglion enhancement was noted on magnetic resonance imaging in several patients. Treatment with mebendazole combined with glucocorticosteroids appeared to shorten the course of the infection, but not the number of relapses. The eosinophil count fell to normal within 3 months, but IgE levels remained elevated for as long as 6 months. All patients recovered with minimal neurologic sequelae. CONCLUSION: Eosinophilic meningitis caused by A. cantonensis should be considered in patients who have headache or central nervous system manifestations after eating raw snails.

Adult↗

Long-term results of duodenectomy with highly selective vagotomy in the treatment of complicated duodenal ulcers.

BACKGROUND: Highly selective vagotomy and complete circular or partial duodenectomy have been applied to complicated duodenal ulcer for many years. These procedures seem to provide better clinical results than truncal vagotomy and antrectomy. METHODS: A retrospective analysis was conducted of 120 patients with complicated duodenal ulcer who underwent surgical treatment between 1986 and 1999. Patients with obstruction were treated with either circular complete (17) or partial duodenectomy (3) combined with highly selective vagotomy or truncal vagotomy and antrectomy (37). Those with perforation were treated primarily with highly selective vagotomy and partial duodenectomy, highly selective vagotomy alone, or truncal vagotomy and pyloroplasty. Every patient was followed up either by a clinic visit (75%) or questionnaire to determine the presence of ulcer pain, dumping, diarrhea, vomiting, weight loss, and Visick grade. RESULTS: Long-term follow-up of patients treated with duodenectomy and highly selective vagotomy for obstruction showed that 94% had sustained weight gain whereas more than half of those treated with truncal vagotomy and antrectomy had weight loss. In patients with perforation, duodenectomy and highly selective vagotomy offered no advantage over highly selective vagotomy alone. CONCLUSIONS: Highly selective vagotomy and complete circular or partial duodenectomy provide fewer sequelae and better weight gain long term than truncal vagotomy and antrectomy for patients with obstructing duodenal ulcers.

Adolescent↗

Soluble Kit receptor blocks stem cell factor bioactivity in vitro.

Stem cell factor (SCF) is a growth factor that promotes the survival, proliferation, and differentiation of hematopoietic cells. SCF and its receptor, Kit, are normally present in both cell surface and soluble forms. Both forms of Kit can bind SCF. However, the function of soluble Kit is unknown. In order to determine if soluble Kit can modulate SCF activity, we produced a fusion protein, Kit-Fc, comprised of the extracellular domain of murine Kit and the Fc portion of human IgG(1) and investigated its ability to bind 125I-SCF and to inhibit SCF-stimulated hematopoietic colony growth in vitro. Stable cell lines expressing Kit-Fc were generated and Kit-Fc was purified to greater than 95% purity. Scatchard analysis demonstrated that Kit-Fc binds iodinated SCF with high affinity (Kd 570 pM). Kit-Fc also bound to transmembrane SCF displayed on the surface of fibroblasts. The murine mast cell line IC2 was engineered to express murine Kit on the cell surface and was demonstrated to proliferate in the presence of SCF. Kit-Fc completely blocked SCF-stimulated proliferation of IC2-Kit cells, but not IL-3-stimulated growth of IC2-Kit cells, demonstrating the specificity of Kit-Fc. We investigated the ability of Kit-Fc to block SCF-stimulated murine hematopoietic colony growth. Kit-Fc blocked SCF-stimulated erythroid colony growth as effectively as a neutralizing anti-Kit monoclonal antibody, ACK2, but did not block erythropoietin-stimulated erythroid colony growth. Likewise, Kit-Fc blocked SCF-stimulated myeloid colony growth as effectively as ACK2 antibody, but did not block IL-3- or GM-CSF-stimulated myeloid colony growth. These results indicate that a form of soluble Kit binds SCF with high affinity, and can specifically block the ability of SCF to stimulate hematopoietic colony growth, suggesting that one function of soluble Kit may be to modulate SCF bioactivity.

3T3 Cells↗