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Biomedical subjects

L Y Chen

Publications and source records attributed to L Y Chen.

At least 19 recordsLinked to original sources
PubMed2003

A novel pathway to enzyme deactivation: the cutinase model.

Cutinase in aqueous solution at pH 4.5 deactivates following a parallel pathway. At 53 degrees C, 88% of the cutinase molecules are in the unfolded conformation, which can aggregate with a reaction order of 3 if the protein concentration is high (>/=12 microM). The aggregates show a sixfold increase in size as determined by dynamic light scattering. This aggregation process is the first phase observed during a deactivation experiment; however, after significant cutinase depletion and maturation of the aggregates, a first-order step starts to dominate and a second phase independent of the protein concentration is observed. Kinetic partitioning between aggregation and first-order irreversible changes of the unfolded conformation can occur during enzyme deactivation when the equilibrium between the native and the unfolded conformation is shifted and kept toward the unfolded conformation.

Carboxylic Ester Hydrolases↗
PubMed2002

Finding transition paths and rate coefficients through accelerated Langevin dynamics.

We present a technique to resolve the rare event problem for a Langevin equation describing a system with thermally activated transitions. A transition event within a given time interval (0,t(f)) can be described by a transition path that has an activation part during (0,t(M)) and a deactivation part during (t(M),t(f))(0<t(M)<t(f)). The activation path is governed by a Langevin equation with negative friction while the deactivation path by the standard Langevin equation with positive friction. Each transition path carries a given statistical weight from which rate constants and related physical quantities can be obtained as averages over all possible paths. We demonstrate how this technique can be used to calculate activation rates of a particle in a two dimensional potential for a wide range of temperatures where standard molecular dynamics techniques are inefficient.

Journal Article↗
PubMed2001

Psychiatric morbidity and post-traumatic symptoms among survivors in the early stage following the 1999 earthquake in Taiwan.

This study reports the clinical psychiatric presentations and post-traumatic symptoms among 525 survivors at Yu-Chyr District in Nantou County who sought psychiatric service in the first month following the devastating earthquake that struck the central area of Taiwan. All subjects received psychiatric interviews and assessments using the 12-item Chinese Health Questionnaire (CHQ-12) and a checklist for post-traumatic symptoms. The most common psychiatric symptoms reported were insomnia, palpitations, nervousness, and dizziness with headache. Eleven percent of the subjects reported having thought of death or having suicidal ideation. The mean score on the CHQ-12 was 6.43 (S.D.=2.89). The rate of probable psychiatric morbidity as defined by a CHQ-12 score > or =3 was 89.9%. Post-traumatic symptoms were very prevalent, particularly symptoms of re-experiencing the earthquake and hyper-arousal. Factors significantly associated with high psychiatric morbidity were being female, serious destruction of property and house, and personality characteristics of nervousness and obsessiveness. Findings of this study suggest that early psychiatric intervention, including pharmacological treatment for acute stress disorder, is indicated during the early stages following a disastrous earthquake.

Adult↗
PubMed2001

[Influence of human cytomegalovirus infection on the expression of HOXB5, HOXB6, HOXB7, and HOXB8 genes in gliomaous cells].

OBJECTIVE: The expressions of HOXB5, HOXB6, HOXB7, and HOXB8 genes of U251 cell infected by human cytomegalovirus and/or treated with all trans-retinoic acid(ATRA) were detected by semi-quantitative RT-PCR. The results were that U251 cell did not express HOXB5, HOXB6, and HOXB8 but expressed HOXB7 without infecting HCMV and/or being treated with ATRA. After infected with HCMV and/or treated with ATRA, the expressions of HOXB7 and HOXB8 were up-regulated and the expression of HOXB7 lasted the fourth generation, while HOXB8 was up-regulated only in the second generation. These results indicate that the abnormal expression of HOXB gene induced by HCMV may play an important role in the maldevelopment.

Brain Neoplasms↗
PubMed2001

Chemoattractant-stimulated NF-kappaB activation is dependent on the low molecular weight GTPase RhoA.

Chemoattractants bind to seven transmembrane-spanning, G-protein-coupled receptors on monocytes and neutrophils and induce a variety of functional responses, including activation of the transcription factor NF-kappaB. The signaling mechanisms utilized by chemoattractants to activate NF-kappaB in human peripheral blood monocytes are poorly defined. We previously demonstrated that fMet-Leu-Phe (fMLP) stimulates NF-kappaB activation, and this function of fMLP requires phosphatidylinositol 3-kinase (PI3K). Here we present evidence that fMLP activates RhoA and that fMLP-induced NF-kappaB activation requires this small GTPase. Stimulation of monocytes with fMLP rapidly activated RhoA as well as NF-kappaB, and their activation was markedly reduced by pertussis toxin treatment. Pretreatment of monocyte with a RhoA inhibitor, C3 transferase from Clostridium botulinum, effectively blocked fMLP-induced NF-kappaB activation as well as interleukin-1beta gene expression. A dominant negative form of RhoA (T19N) also inhibited fMLP-stimulated reporter gene expression in a kappaB-dependent manner. Cotransfection of the monocytic THP1 cells with a constitutively active form of RhoA (Q63L) with the promoter reporter plasmid results in a marked increase in NF-kappaB-mediated reporter gene expression. Furthermore, the PI3K inhibitors wortmannin and LY294002 block RhoA activation induced by fMLP. These results demonstrate that low molecular weight GTPase RhoA is a novel signal transducer for fMLP-induced NF-kappaB activation and Galpha(i) or Galpha(o) class of heterotrimeric G proteins likely mediate RhoA activation via PI3K in human peripheral blood monocytes.

Base Sequence↗
PubMed2001

[Listeria monocytogenes induces thymocyte apoptosis in mice].

The murine thymocyte apoptosis induced by Listeria monocytogenes(LM) was detected with morphology, FCM, and DNA electrophoresis. The results were that LM elicited typical morphological changes of thymocyte apoptosis; the typical apoptosis peak was displayed with FCM, and typical "ladder pattern" with agarose gel electrophoresis. The apoptotic cells were found at 8 h after the mice had infected LM and reached climax at 48 h. The thymus weight significantly reduced at 16 h, and reached the lowest at 48 h after the mice had infected LM. The percentage of apoptotic cells was raised with the increasing of LM. These results suggest that LM induces thymocyte apoptosis in dose- and time-dependent manner.

Animals↗
PubMed2001

[Influence of human cytomegalovirus infection on the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung cells].

OBJECTIVE: To study the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung (HEL) cells and influence of human cytomegalovirus (HCMV) infection on the expressions of these genes. METHOD: The expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes were detected with semi-quantitative RT-PCR method. RESULTS: 1. HEL cells expressed HOXB5 and HOXB6 genes, but they did not express HOXB1 and HOXB9 genes. 2. After HCMV infection, HEL cell was induced to express HOXB9 gene, as the expression of HOXB6 gene was elevated. The expression of HOXB5 gene had no significant change. HOXB1 was still not expressed. 3. Treated with all-trans retinoic acid (ATRA), the expression of HOXB9 gene in HEL cells infected by HCMV was significantly increased. But during the advanced infect period, the expression of HOXB6 was significantly decreased. CONCLUSION: HCMV can induce abnormal expressions of HOXB6 and HOXB9 genes, which may play an important role in HCMV-induced abnormal embryogenesis.

Cells, Cultured↗
PubMed2001

[Study on molecular epidemiology of HCMV infection in mothers and their newborns in Changsha].

The detection of HCMV-IgG, IgM in plasma by ELISA and DNA in plasma and P(C) BMCs from mothers and their newborns by PCR was carried out. Positive HCMV-IgG, HCMV-IgM, P(C)BMCs HCMV DNA and plasma HCMV DNA were demonstrated in 119(95.2%), 7(5.6%), 27(21.6%) and 14(11.2%) in 125 mothers and 117(93.6%), 1(0.8%), 18(14.4%) and 7(5.6%) in their mewborns, respectively. There were significant differences among the positive rate of newborn's HCMV-IgM, plasma HCMV DNA and CBMCs HCMV DNA (P < 0.05), but the difference between HCMV-IgM and plasma HCMV DNA had no statistic significance (P > 0.05). This study suggests that HCMV-IgG and CBMCs HCMV PCR are preferable epidemiological index of HCMV infection of pregnant women and newborns respectively.

Adult↗
PubMed2001

Surgical correction of buried penis: a review of 60 cases.

BACKGROUND/PURPOSE: The authors reviewed 60 cases of buried penis, treated in a single institution with postoperative follow-up for at least 6 months. The review was aimed at providing information that might help to optimize the results in future surgical correction of this uncommon, but not rare, congenital disorder. METHODS: From January 1, 1989 to December 31, 1998, 62 boys with buried penis were treated with 1 of the following procedures: group 1A (n = 6), preputial unfurling alone; group 1B (n = 8), modified preputial unfurling; group 1C (n = 12), penoplasty devised by the first author with preservation of the preputial skin; and group 2 (n = 36), penoplasty with trimming of the inner preputial skin. Postoperative follow-up of more than 6 months after operation was achieved in 60 of 62 patients for a total of 25 patients in group 1 (A through C) and 35 patients in group 2. RESULTS: Recurrent buried penis developed in 8 of 60 patients (13%), and redundant penile skin with or without lymphedema occurred in 18 (30%). The complications occurred in 18 of 25 patients (72%) in group 1 (A through C) but in only 8 of 35 (23%) in group 2. The difference was significant (P =.001). Most of the complications were mild and acceptable. A second procedure was required in 5 of the 25 patients in group 1 but in none of group 2. The second procedures were required to correct recurrent buried penis in 1 and to resect excess redundant penile skin in 4. CONCLUSION: The superior results achieved in group 2 suggest that in addition to penoplasty with adequate fixation of the unfurling prepuce, resection of excess inner preputial skin is required to achieve an optimal outcome in children requiring surgical correction of buried penis.

Child↗
PubMed2001

Involvement of the XpsN protein in formation of the XpsL-xpsM complex in Xanthomonas campestris pv. campestris type II secretion apparatus.

The xps gene cluster is required for the second step of type II protein secretion in Xanthomonas campestris pv. campestris. Deletion of the entire gene cluster caused accumulation of secreted proteins in the periplasm. By analyzing protein abundance in the chromosomal mutant strains, we observed mutual dependence for normal steady-state levels between the XpsL and the XpsM proteins. The XpsL protein was undetectable in total lysate prepared from the xpsM mutant strain, and vice versa. Introduction of the wild-type xpsM gene carried on a plasmid into the xpsM mutant strain was sufficient for reappearance of the XpsL protein, and vice versa. Moreover, both XpsL and XpsM proteins were undetectable in the xpsN mutant strain. They were recovered either by reintroducing the wild-type xpsN gene or by introducing extra copies of wild-type xpsL or xpsM individually. Overproduction of wild-type XpsL and -M proteins simultaneously, but not separately, in the wild-type strain of X. campestris pv. campestris caused inhibition of secretion. Complementation of an xpsL or xpsM mutant strain with a plasmid-borne wild-type gene was inhibited by coexpression of XpsL and XpsM. The presence of the xpsN gene on the plasmid along with the xpsL and the xpsM genes caused more severe inhibition in both cases. Furthermore, complementation of the xpsN mutant strain was also inhibited. In both the wild-type strain and a strain with the xps gene cluster deleted (XC17433), carrying pCPP-LMN, which encodes all three proteins, each protein coprecipitated with the other two upon immunoprecipitation. Expression of pairwise combinations of the three proteins in XC17433 revealed that the XpsL-XpsM and XpsM-XpsN pairs still coprecipitated, whereas the XpsL-XpsN pair no longer coprecipitated.

Bacterial Proteins↗
PubMed2001

A common precursor for the three subunits of L-glutamate oxidase encoded by gox gene from Streptomyces platensis NTU3304.

A segment of DNA containing the L-glutamate oxidase (gox) gene from Streptomyces platensis NTU3304 was cloned. The entire nucleotide sequence of the protein-coding portion consisting of 2130 bp (710 codons, including AUG and UGA) of the cloned DNA fragment was determined. The gox gene contained only one open reading frame (ORF) which coded for a 78-kDa polypeptide, the precursor of active extracellular Gox. Mature Gox is composed of three subunits, designated as alpha, beta, and gamma, with molecular masses of 39, 19, and 16 kDa, respectively. Analyses of the N-terminal amino acid sequences of the subunits revealed that the order of subunits in the precursor polypeptide encoded by the ORF, from N-terminus to C-terminus, is alpha-gamma-beta. The presence of the flavin adenine dinucleotide (FAD)-binding motif place Gox as a member of the flavoenzyme family. Furthermore, a negative effect of glucose on the biosynthesis of Gox was observed when it was used as carbon source.

Amino Acid Oxidoreductases↗
PubMed2000

Benzylidene ketal derivatives as M2 muscarinic receptor antagonists.

Benzylidene ketal derivatives were investigated as selective M2 receptor antagonists for the treatment of Alzheimer's disease. Compound 10 was discovered to have subnanomolar M2 receptor affinity and 100-fold selectivity against other muscarinic receptors. Also, 10 demonstrated in vivo efficacy in rodent models of muscarinic activity and cognition.

Acetylcholine↗
PubMed2000

Structural requirements for the stability and microsomal transport activity of the human glucose 6-phosphate transporter.

Deficiencies in glucose 6-phosphate (G6P) transporter (G6PT), a 10-helical endoplasmic reticulum transmembrane protein of 429 amino acids, cause glycogen storage disease type 1b. To date, only three missense mutations in G6PT have been shown to abolish microsomal G6P transport activity. Here, we report the results of structure-function studies on human G6PT and demonstrate that 15 missense mutations and a codon deletion (delta F93) mutation abolish microsomal G6P uptake activity and that two splicing mutations cause exon skipping. While most missense mutants support the synthesis of G6PT protein similar to that of the wild-type transporter, immunoblot analysis shows that G20D, delta F93, and I278N mutations, located in helix 1, 2, and 6, respectively, destabilize the G6PT. Further, we demonstrate that G6PT mutants lacking an intact helix 10 are misfolded and undergo degradation within cells. Moreover, amino acids 415-417 in the cytoplasmic tail of the carboxyl-domain, extending from helix 10, also play a critical role in the correct folding of the transporter. However, the last 12 amino acids of the cytoplasmic tail play no essential role(s) in functional integrity of the G6PT. Our results, for the first time, elucidate the structural requirements for the stability and transport activity of the G6PT protein.

Amino Acid Sequence↗
PubMed2000

[Influence of human cytomegalovirus infection on the expression of HOX genes in human embryo lung cells].

The expressions of HOX genes in human embryo lung (HEL) cells were detected with semi-quantitative RT-PCR method. The results were that HEL cells expressed HOXB7 gene and its expression increased after human cytomegalovirus (HCMV) infection. The level of expression reached maximum at 48 h after HCMV infection. Treated with all-trans retinoic acid (ATRA), the expression of HOXB7 in HEL cells infected by HCMV was significantly increased. The results suggest that the abnormal expression of HOX genes induced by HCMV might possibly play a role in virus-induced abnormal embryogenesis.

Cells, Cultured↗
PubMed2000

Diphenylsulfone muscarinic antagonists: piperidine derivatives with high M2 selectivity and improved potency.

Piperidine analogues of our previously described piperazine muscarinic antagonists are described. Piperidine analogues show a distinct structure-activity relationship (SAR) that differs from comparable piperazines. Compounds with high selectivity and improved potency for the M2 receptor have been identified. The lead compound, 12b, increases acetylcholine release in vivo. Compounds of this class may be useful for the treatment of cognitive disorders such as Alzheimer's disease (AD).

Acetylcholine↗
PubMed2000

Retinoic acid-mediated G1 arrest is associated with induction of p27(Kip1) and inhibition of cyclin-dependent kinase 3 in human lung squamous carcinoma CH27 cells.

Retinoids are promising agents for the prevention and treatment of several human malignancies including lung cancer. In this study, the effect of retinoic acid (RA) on cell growth and the mechanism of growth modulation were examined in human lung squamous carcinoma CH27 cells. Here we report that RA mediated the dose- and time-dependent growth arrest in G1 phase, accompanied by the up-regulation of p27(Kip1) and the down-regulation of the cyclin-dependent kinase 3 (Cdk3) and p21(CIP1/Waf1) proteins. Furthermore, RA-induced growth arrest of CH27 cells was also associated with increased retinoic acid receptor beta (RARbeta) and reduced c-Myc expression. However, RA had no effect on the levels of cyclins A, D1, D3, E, or H, or on Cdk2, Cdk4, Cdk5, CDk6, Cdk7, p16(Ink4A), p15(Ink4B), p53, or pRb proteins in CH27 cells. Evaluation of the kinase activity of cyclin-Cdk complexes showed that RA increases p27(Kip1) expression in CH27 cells leading to markedly reduced cyclin A/Cdk2 kinase activity and slightly reduced cyclin E/Cdk2 kinase activity, with no effect on cyclin D/Cdk4 and cyclin D/Cdk6 activities. Moreover, coincident with the decrease in kinase activity was a drastic increase in cyclin A-bound p27(Kip1). These results suggest that increases in the levels of p27(Kip1) and its binding to cyclin A, as well as reduction of Cdk3 protein expression, are strong candidates for the cell cycle regulator that prevents the entry into the S phase in RA-treated CH27 cells, with prolongation of G1 phase and inhibition of DNA synthesis.

CDC2-CDC28 Kinases↗
PubMed2000

Cellular release of [18F]2-fluoro-2-deoxyglucose as a function of the glucose-6-phosphatase enzyme system.

[(18)F]-2-Fluoro-2-deoxyglucose (FDG) is a glucose analog currently utilized for positron emission tomography imaging studies in humans. FDG taken up by the liver is rapidly released. This property is attributed to elevated glucose-6-phosphatase (Glc-6-Pase) activity. To characterize this issue we studied the relationship between Glc-6-Pase activity and FDG release kinetics in a cell culture system. We overexpressed the Glc-6-Pase catalytic unit in a Glc-6-Pase-deficient mouse hepatocyte (Ho-15) and in A431 tumor cell lines. Glc-6-Pase enzyme activity and FDG release rates were determined in cells transfected with the Glc-6-Pase gene (Ho-15-D3 and A431-AC3), in mock-transfected cells of both cell lines, and in wild-type mouse hepatocytes (WT10) as control. Although the highest level of Glc-6-Pase activity was measured in A431-AC3, Ho-15-D3 cells showed much faster FDG release rates. The faster FDG release correlated with the level of glucose 6-phosphate transporter (Glc-6-PT) mRNA, which was found to be expressed at higher levels in Ho-15 compared with A431 cells. Overexpression of Glc-6-PT in A431-AC3 produced a dramatic increase in FDG release compared with control cells. This study gives the first direct evidence that activity of the Glc-6-Pase complex can be quantified in vivo by measuring FDG release. Adequate levels of Glc-6-Pase catalytic unit and Glc-6-PT are required for this function. FDG-positron emission tomography may be utilized to evaluate functional status of the Glc-6-Pase complex.

Animals↗