Biomedical subjects

Ching-Wen Chang

Publications and source records attributed to Ching-Wen Chang.

9 recordsLinked to original sources

PubMed2005

Transcriptome analysis in blastocyst hatching by cDNA microarray.

BACKGROUND: Hatching is an important process for early embryo development, differentiation and implantation. However, little is known about its regulatory mechanisms. By integrating the technologies of RNA amplification and cDNA microarrays, it has become possible to study the gene expression profile at this critical stage. METHODS: Pre-hatched and hatched ICR mouse embryos (25 blastocysts in each group were used in the triplicate experiments) were collected for RNA extraction, amplification, and microarray analysis (the mouse cDNA microarray, 6144 genes, including expressed sequence tags). RESULTS: According to cDNA microarray data, we have identified 85 genes that were expressed at a higher level in hatched blastocyst than in pre-hatched blastocysts. In this study, 47 hatching-related candidate genes were verified via re-sequencing. Some of these genes have been selected and confirmed by real-time quantitative RT-PCR. These hatching-specific genes were also expressed at a lower level in the delayed growth embryos (morula or blastocyst without hatching at day 6 post hCG). These genes included: cell adhesion and migration molecules [E-cadherin, neuronal cell adhesion molecule (NCAM), lectin, galactose binding, soluble 7 (Lgals7), vanin 3 and biglycan], epigenetic regulators (Dnmt1, and SIN3 yeast homolog A), stress response regulators (heme oxygenase 1) and immunoresponse regulators [interleukin (IL)-2-inducible T-cell kinase, IL-4R, interferon-gamma receptor 2, and neurotrophin]. The immunostaining of E-cadherin and NCAM showed strong and specific localization in hatched blastocyst. CONCLUSIONS: This work provides important information for studying the mechanisms of blastocyst hatching and implantation. These hatching-specific genes may have potential as new drug targets for controlling fertility.

Animals↗

PubMed2005

FBW2 targets GCMa to the ubiquitin-proteasome degradation system.

The GCM proteins GCMa/1 and GCMb/2 are novel zinc-containing transcription factors critical for glial cell differentiation in fly and for placental as well as parathyroid gland development in mouse. Previous pulse-chase experiments have demonstrated differential protein stabilities of GCM proteins with half-lives from approximately 30 min to 2 h (Tuerk, E. E., Schreiber, J., and Wegner, M. (2000) J. Biol. Chem. 275, 4774-4782). However, little is known about the machinery that controls GCM protein degradation. Here, we report the identification of an SCF complex as the GCM ubiquitin-protein isopeptide ligase (E3) that regulates human GCMa (hGCMa) degradation. We found that SKP1 and CUL1, two key components of the SCF complex, associate with hGCMa in vivo. We further identify the human F-box protein FBW2 (hFBW2) as the substrate recognition subunit in the SCF E3 complex for hGCMa. We show that hFBW2 interacts with hGCMa in a phosphorylation-dependent manner and promotes hGCMa ubiquitination. Supporting a critical role for hFBW2 in hGCMa degradation, knockdown of hFBW2 expression by RNA interference leads to a reduction in hGCMa ubiquitination and a concomitant increase in hGCMa protein stability. Our study identifies the SCF(hFBW2) E3 complex as the key machinery that targets hGCMa to the ubiquitin-proteasome degradation system.

Animals↗

PubMed2004

Identification of a novel microtubule-destabilizing motif in CPAP that binds to tubulin heterodimers and inhibits microtubule assembly.

We have previously identified a new centrosomal protein, centrosomal protein 4.1-associated protein (CPAP), which is associated with the gamma-tubulin complex. Here, we report that CPAP carries a novel microtubule-destabilizing motif that not only inhibits microtubule nucleation from the centrosome but also depolymerizes taxol-stabilized microtubules. Deletion mapping and functional analyses have defined a 112-residue CPAP that is necessary and sufficient for microtubule destabilization. This 112-residue CPAP directly recognizes the plus end of a microtubule and inhibits microtubule nucleation from the centrosome. Biochemical and functional analyses revealed that this 112-residue CPAP also binds to tubulin dimers, resulting in the destabilization of microtubules. Using the tetracycline-controlled system (tet-off), we observed that overexpression of this 112-residue CPAP inhibits cell proliferation and induces apoptosis after G2/M arrest. The possible mechanisms of how this 112-residue motif in CPAP that inhibits microtubule nucleation from the centrosome and disassembles preformed microtubules are discussed.

Amino Acid Motifs↗

PubMed2004

Serum and endometrial markers.

Endometriosis is a benign but aggressive disease. It occurs when shed endometrium from the female reproductive tract grows at a site outside the uterus. The physiological changes in endometriosis-abnormal tissue growth, invasion, and adhesion phenomena-are similar to those seen in tumorous tissues. Although the etiology of endometriosis is not well understood, the disease is widely accepted to result from the ectopic implantation of refluxed menstrual tissues. In addition, immunologic changes, genetic factors, and environmental factors might also affect a woman's susceptibility to develop endometriosis. Thus far, laparoscopic examination is required to confirm the presence of endometriosis; there is no reliable marker for its diagnosis. Many studies are therefore focusing on identifying markers for the diagnosis and follow-up of endometriosis. This chapter provides a systematic review of these studies, including recent findings from our group on the identification of molecules, in serum and/or endometrium, which are associated with the development of endometriosis at different stages. From this research, we hope to be able to suggest how to approach the potential markers. The identification of highly sensitive and specific markers of endometriosis should facilitate the development of accurate and non-invasive techniques for diagnosis and prognosis.

Autoantibodies↗

PubMed2003

Mutational analysis of the KIX domain of CBP reveals residues critical for SREBP binding.

Structure-based mutagenesis was used to probe the binding surface for the activation domain of sterol-responsive element binding protein (SREBP) in the KIX domain of CREB binding protein. A set of conserved residues scattering in the alpha2 helix and the extended C-terminal region of alpha 3 helix in the KIX domain including two arginines previously characterized as a hot spot for cofactor-mediated methylation was shown to be crucial for SREBP-KIX interaction, and was not essential for phosphorylated KID recognition. Therefore, our results suggest the existence of a SREBP binding site formed by positively charged residues in the C-terminal part of the extended alpha 3 helix of the KIX domain distinct from the previously identified phosphorylated KID binding site.

Amino Acid Sequence↗

PubMed2002

Using structural equation model to explore occupational lead exposure pathways.

The aim of the present study was to demonstrate the use of a structural equation model to explore the complicated lead exposure pathways in a lead battery plant. A total of 96 out of 113 assembly workers were recruited in this study. Lead measurements included blood lead, respirable airborne lead, and body surface lead loadings. Latent variables of inadvertent contact of lead on fingers and ingested lead had indirect and direct effects on the blood lead levels, respectively, and so did the variables of respirable airborne lead, smoking, and the working-place clothes being washed. A model chi(22)2 value of 31.1 (P=0.094) for a goodness of fit test indicates that the derived structural equation model appropriately accounted for the variation of blood lead levels. It is concluded that lead loadings on fingers and lips had significant effects on occupational lead uptake, and, after-work hours and break times at work were the critical time periods for inadvertent lead exposure.

Activities of Daily Living↗

PubMed2002

Global analysis of differentially expressed genes in early gestational decidua and chorionic villi using a 9600 human cDNA microarray.

The global gene expression profiles of the decidua and chorionic villi of early human pregnancies were analysed by using cDNA microarray technology. Decidual and villous placental tissues were obtained from first trimester abortus and mRNA was extracted for cDNA microarray analysis. The human cDNA microarray [9600 clones, including known regulatory genes and expressed sequence tags (EST)] with colorimetric detection was used to identify differentially expressed genes between early gestational decidua and villi. According to cDNA microarray analysis, we have identified 641 genes with highly expressed mRNA in both decidua and villi, 49 genes with higher expressions in decidua, and 75 genes with higher expression in chorionic villi. These differentially expressed genes were further grouped into categories by their putative functions, including: cell growth-related factors, hormones/cytokines, cell adhesion molecules, signal transduction molecules, apoptosis-related factors, cytoskeleton/extracellular matrix proteins, and EST. Immunohistochemical stainings of cathepsin L, leukaemia inhibitory factor-receptor, and proliferative cell nuclear antigen showed results consistent with the microarray data. Identification of the differentially expressed genes between decidua and villi by microarray provide a global profiling of the gene expression pattern. This work adds to our understanding of placentation by reporting the gene expression profiles during first trimester human pregnancies using cDNA microarray.

Adult↗

PubMedDate not supplied

Fungi genus and concentration in the air of onion fields and their opportunistic action related to mycotic keratitis.

The authors determined fungi concentrations and genera from onion fields located in monsoon and nonmonsoon areas of Taiwan, as well as concentrations from a county housing location, which served as the comparison site. The authors explored the relationship(s) among onion harvesting, monsoon characteristics, airborne fungal levels, and mycotic eye infection. Airborne fungal levels in the onion field in the nonmonsoon area were the highest, followed by decreasing levels in the monsoon area and control sites, regardless of the sampling periods. The geometric mean of airborne fungal concentrations was the highest during onion harvesting. Cladosporium was the most dominant fungus during pre- and postharvest seasons, whereas Aspergillus, Fusraium, Acremonium, Alternaria, Penicillium, Monilia, Mycelia, and Rhizopus were increased significantly during harvesting. Although fungal concentrations in monsoon onion fields were lower than concentrations measured in the nonmonsoon area, the invasion of dangerous fungal pathogens following eye trauma by airborne sharp exotics (e.g., onion flakes) dispersed in high wind may account for the increase in corneal ulcer cases that occur in the monsoon area.

Adult↗

PubMedDate not supplied

Suspended onion particles and potential corneal injury in onion harvesters.

The authors suspected that suspended onion particles contributed to corneal ulcers in onion harvesters in southern Taiwan. In the present study, the authors used manikins to study suspended onion particles in fields in an effort to simulate typical conditions experienced by onion harvesters. An animal eye-exposure simulation study was also performed by the authors, who impacted suspended soil grains or onion particles onto the corneas of guinea pigs via aerosol generated from the Palas dispersion nozzle. The average size of 25.9 pm for suspended particles collected during the digging of onions was the largest one of those for various harvesting activities. Some onion skin flakes were found in samples obtained from gathering and packing activities; the typical flake size was approximately 3.5 x 2.5 mm2. The results of the animal study indicated that the size of soil grains has a demonstrable effect on the severity of corneal injury (p = .009). With respect to onion skin flakes, wind velocity was also associated significantly with the occurrence of corneal injury (p = .0004). A wind velocity threshold of 7 m/sec is recommended for the maintenance of safety, and if the wind speed exceeds this threshold level, workers should not engage in harvesting activities. Furthermore, use of appropriately designed goggles is necessary for the protection of onion harvesters who work in high-wind conditions.

Agricultural Workers' Diseases↗