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Biomedical subjects

C K Chou

Publications and source records attributed to C K Chou.

At least 55 records · Page 3Linked to original sources
PubMed1998

Phosphatidylinositol 3-kinase is required for the regulation of hepatitis B surface antigen production and mitogen-activated protein kinase activation by insulin but not by TPA.

Insulin suppresses hepatitis B surface antigen (HBsAg) gene expression and stimulates cell proliferation in human hepatoma Hep3B cells. 12-O-tetradecanoyl phorbol-13-acetate, TPA, has been demonstrated to mimic insulin actions in these cells. We examined the role of phosphatidylinositol 3-kinase (PI 3-kinase) in the signaling pathways of insulin and TPA towards these two biological phenomena in Hep3B cells. The pre-treatment of 5 microM of wortmannin diminished insulin suppressed HBsAg production and completely abolished insulin stimulated cell proliferation. However, wortmannin had no effect on TPA actions in both HBsAg suppression and cell growth stimulation. We further investigated the effect of wortmannin in mitogen-activated protein kinases (MAPKs) activation induced by insulin or TPA. After the pretreatment of wortmannin, insulin activated MAPKs was completely blocked, but TPA was still capable to activate MAPKs. These results suggest that PI 3-kinase is involved in insulin actions but not in TPA effects, and allow us to dissociate the signaling pathways of insulin and TPA in human hepatoma Hep3B cells.

Androstadienes↗
PubMed1998

Apoptotic signal of Fas is not mediated by ceramide.

Ceramide has been suggested as the secondary messenger mediating the apoptotic signal for Fas engagement. By using different inhibitors, we demonstrated here that ceramide is unlikely a mediator of Fas-initiated apoptosis. First, cAMP prevented cell death induced by ceramide but not by Fas. Second, ceramide-triggered, but not Fas-triggered, apoptosis was antagonized by the free radical scavenger C60. Third, the metal chelator pyrrolidinedithiocarbamate suppressed ceramide-initiated DNA fragmentation but had no effect on the Fas-induced cell death. Fourth, the SAPK/ERK kinase dominant negative mutant, which attenuated ceramide-induced cell death, did not prevent Fas-induced apoptosis. Finally, activation of NF-kappaB inhibited ceramide-induced but not Fas-initiated apoptosis. The fact that many antagonists of ceramide-induced apoptosis could not suppress Fas-mediated cell death clearly indicates that ceramide is not the mediator for Fas-initiated apoptotic signal.

Apoptosis↗
PubMed1998

A conserved sequence block in murine and human T cell receptor (TCR) Jalpha region is a composite element that enhances TCR alpha enhancer activity and binds multiple nuclear factors.

A conserved sequence block (CSB) located in a noncoding region of the mouse and human TCR alpha/delta loci, showing six differences over 125 nucleotide positions (95% similar), was subjected to detailed analyses in this study. Transient transfection results showed that the CSB-containing element in conjunction with the TCR alpha enhancer up-regulated the alpha enhancer activity, whereas no enhancer activity was detected when CSB alone was assayed. In vitro occupancy analyses of CSB by nuclear factors reveal the existence of an unexpectedly intricate network of CSB-protein and protein-protein interactions. Lymphoid-specific as well as T-lineage-specific nuclear factors are involved to differentially form CSB-bound complexes in extracts of various tissues and cell lines. Liver was shown to contain factor(s) sequestering thymic CSB-binding factors. Furthermore, the putative binding sites for transcription factors known to be important for lymphoid-lineage development are present in CSB and are targeted by nuclear factors. On the basis of these results, we propose that the CSB element may play a role in shaping the chromatin structure by which the accessibility of TCR alpha/delta loci to the recombinase complex and/or to the transcriptional apparatus can be controlled.

Animals↗
PubMed1998

Genomic organization and characterization of the promoter region of the round-spotted pufferfish (Tetraodon fluviatilis) JAK1 kinase gene.

Seventeen kilobases of genomic DNA containing the promoter and the coding region of the round-spotted pufferfish JAK1 gene was isolated and completely sequenced. This gene consists of 25 exons and 24 introns spanning about 13.5 kb, compared to > 30kb in carp JAK1 gene. Primer extension analysis revealed one transcription initiation site which was 376 bp upstream of the translation initiation site. The sequence of the 2.9 kb region upstream of the transcription initiation site contains numerous potential binding sites for transcription factors including HNF-5, GCF, Sp1, CRE, AP2, GATA, GAGA, E2A, p53, and NF-IL6. When this region was placed upstream of the chloramphenicol acetyltransferase (CAT) reporter gene and transfected into a carp CF cell line, it could drive the synthesis of CAT enzyme three times more efficiently than could the common carp JAK1 promoter.

Animals↗
PubMed1998

Hepatocyte growth factor stimulates the growth and activates mitogen-activated protein kinase in human hepatoma cells.

Hepatocyte growth factor (HGF) is a potent mitogen for hepatocytes and various epithelial cells. Unexpectedly, it has been reported to inhibit the growth of hepatoma cells in vitro. To clarify this phenomenon, we examined the effects of recombinant baculovirus-expressed HGF on the growth of 6 human hepatoma cell lines. The growth of Hep3B and HepG2 cells was markedly stimulated to 1.8- and 1.7-fold, respectively, PLC/PRF/5 to 1.4-fold, and SK-Hep-1 to 1.2-fold in a dose-dependent manner under HGF concentrations below 20 ng/ml. Neither HuH-7 nor HCC36 were affected. None of these cells were inhibited. All these cells expressed c-Met, the membrane receptor for HGF, and their c-Met would be activated to be phosphorylated upon addition of HGF. They also contained the ERK2 subgroup of mitogen-activated protein kinases (MAPKs). When HGF was added, their ERK2 would also be phosphorylated. The extent of ERK2 phosphorylation was partially correlated to their growth response to HGF. In conclusion, HGF could stimulate the growth of certain human hepatoma cells, probably through activation of c-Met and MAPKs.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1998

Bcl-2 blocks apoptotic signal of transforming growth factor-beta in human hepatoma cells.

Transforming growth factor-beta (TGF-beta) has been shown to induce apoptosis on normal hepatocytes and hepatoma cells both in vitro and in vivo. However, how the TGF-beta induces apoptosis is still not clear. We examined the expression of anti-apoptosis proteins and sensitivity to TGF-beta in three well differentiated human hepatoma cell lines. Two TGF-beta sensitive cell lines Hep3B and HuH7 totally lacked Bcl-2. In contrast, the TGF-beta resistant HepG2 cells expressed a substantial amount of Bcl-2. All three cell lines expressed equal amounts of Bcl-X(L), Bcl-X(S) and Bax. Overexpression of Bcl-2 in Hep3B and HuH7 cells protected them from TGF-beta-induced apoptosis. TGF-beta treatment increased intracellular peroxide production and suppressed the expression of glutathione-S-transferase in the Hep3B cells, and these effects were partially suppressed by the overexpression of Bcl-2. These results suggest that Bcl-2 may protect cell from TGF-beta-F-induced apoptosis by interfering TGF-beta generated signals leading to induce reactive oxygen species production.

Apoptosis↗
PubMed1998

Isolation and identification of novel protein kinase genes from the round-spotted pufferfish (Tetraodon fluviatilis) genomic DNA.

The round-spotted pufferfish Tetraodon fluviatilis has a genome size of 380 Mb which is slightly smaller than that of another pufferfish, Fugu rubripes rubripes (Fugu). Due to their compact genome and small introns, both pufferfishes have been proposed as model organisms for genome studies. In this study, we have used genomic DNA as template to perform PCR to screen for protein kinase (pk) genes. Forty-one T. fluviatilis pk genes encoding 7 receptor tyrosine kinases, 14 nonreceptor tyrosine kinases, 16 serine/threonine kinases, 1 dual kinase and 3 novel kinases have been identified. The success of this approach depends on the size and location of the introns. Most of the identified pk gene fragments contain introns, ranging from 71 to 300 bp, with an average of 120 bp. It is noteworthy that the intron/exon boundaries of certain genes which belong to the same family are identical. We also analyzed by specific RT-PCR primers the expression profile of those 3 novel genes as well as some selected pk genes in a variety of tissues. We found that erbB3, pku a, mrk, CaMK I, CaMKIIgamma, and two novel kinase genes (133 and 3-26) are expressed in all tissues examined. However, the novel clone 146 is strongly expressed in the brain and weakly in the intestine, kidney and heart.

Amino Acid Sequence↗
PubMed1998

From mosquito to man: identification of a novel protein kinase, HsHPK, which is highly expressed in human hepatoma tissues.

Protein kinases play an important role in the signaling pathway of growth factors in most of the higher organisms. During the study of protein kinase profiles of mosquitoes using RT-PCR and degenerate primers for consensus catalytic domain motifs to amplify protein kinase genes, we have noticed that a novel mosquito kinase, AaPK-38, shares a stretch of amino acids identical to the corresponding domain in Tousled gene of Arabidopsis thaliana that is required for leaf and flower development. A 2.1-kb cDNA encoding human HsHPK gene, which is a homolog of AaPK-38, was isolated from human testis cDNA library. This cDNA contains an open reading frame of 563 amino acids, with a complete kinase domain in its carboxyl terminus. The expressed Flag-tagged HsHPK was shown to have kinase activity based on in vitro autophosphorylation. Northern blot analysis revealed that human HsHPK mRNA is most abundant in testes, much less in heart and skeletal muscle and almost undetectable in liver and lung. Finally, we found that the expression of HsHPK in 4 out of 6 human hepatoma tissues is much higher than that in the adjacent normal counterpart. This result suggests HsHPK may play a role in the development of human hepatoma.

Amino Acid Sequence↗
PubMed1998

Mechanical properties of laser-welded cast titanium joints under different conditions.

STATEMENT OF THE PROBLEM: Data on optimal conditions for laser welding commercial pure cast titanium used in dental restorations are sparse and not well established. PURPOSE: The mechanical properties of laser-welded cast commercially pure titanium under different laser welding conditions were evaluated to find the optimal parameters in terms of duration and voltage (energy level). MATERIAL AND METHODS: Fifty-seven cast pure titanium specimens according to ISO specification 6871 were divided into nine experimental groups and one control group. The sectioned titanium bars were laser-welded under different duration (8, 10, and 12 ms) and energy (290, 300, and 310 V) levels and evaluated for ranges of ultimate tensile strength (374 to 562 MPa), 0.2% yield strength (206 to 338 MPa), and percentage elongation (2.49% to 10.58%). RESULTS: Under a suitable laser-welding setting parameter the ultimate tensile strength and 0.2% yield strength of experimental groups were weaker than the unsectioned control specimens (control group ultimate tensile strength 540 +/- 11 MPa, 0.2% yield strength 258 +/- 49 MPa) or too brittle (control group percentage elongation 12.41% +/- 1.83%). Two-way ANOVA, nonparametric tests showed that voltage was the only significant factor for all parameters studied (p < 0.01). Optimal conditions were determined with three-dimensional response curves (305 V, 12 ms ultimate tensile strength; 310 V, 10 ms 0.2% yield strength; 300 V, 12 ms percentage elongation). CONCLUSION: Optimal duration and voltage (energy level) used in laser welding for cast CP grade I titanium bar was superior to or comparable with and produced properties that were superior to or comparable with the unsectioned control specimens.

Analysis of Variance↗
PubMed1998

Endoscopically-assisted adipofascial flap harvest for soft tissue defects of the lower leg.

Five cases of soft tissue defect of the lower leg were reconstructed successfully with the distally based medial adipofascial flap which was harvested with endoscopically-assisted surgery. The size of these flaps ranged from 4.0 x 7.0 cm to 5.0 x 15.0 cm in size. All the flaps had good perfusion and uneventful course postoperatively. The donor site skin was intact except for an additional access-scar 2.5 cm in length. The donor site morbidity of the adipofascial flap was further minimised in this series.

Accidents, Traffic↗
PubMed1998

The distally based lateral adipofascial flap.

The distally based lateral adipofascial flap can be based either on the lowermost perforator or the anterior perforating branch of the peroneal artery avoiding sacrifice of the main peroneal artery. It has been used successfully to resurface soft tissue defects either on the lateral or medial aspect of the lower third of the leg in 13 cases. The size of these flaps varied from 2.0 cm x 5.0 cm to 4.5 cm x 15.0 cm in size. These cases had minimal donor site morbidity and had positive aesthetic results. The advantages were: 1. Choice of either the lowermost perforator or the anterior perforating branch giving a wide arc of rotation; 2. Preservation of the superficial peroneal nerve; 3. Primary closure of the donor site.

Adult↗
PubMed1998

Round-spotted pufferfish (Tetraodon fluviatilis) snf5 gene is oriented in a tail-to-tail manner with the set gene which encodes an inhibitor of protein phosphatase 2A.

The round-spotted pufferfish Tetraodon fluviatilis has a genome size of 380 Mb which is slightly smaller than that of another pufferfish Fugu rubripes rubripes (Fugu). Due to its compact genome and small introns, Fugu has been introduced as a model for genome studies. Recently, the round-spotted pufferfish has also been proposed as a new model for genome studies because of the ease in obtaining material and high-sequence homology to that of Fugu. In this study, we have cloned and characterized the snf5 and set genes from the round-spotted pufferfish. The snf5 gene is composed of 9 exons spanning about 2.9 kb whereas the set gene consists of 8 exons spanning about 2.7 kb. They are linked in a tail-to-tail manner with an intergenic region of about 6.5 kb. So far, the genomic structures of human snf5 and set genes are unknown. Based on our data, the pufferfish SNF5 and SET display high amino acid sequence identity (>90%) with the respective human genes. By primer extension and sequence analysis, we found that putative promoter region of the snf5 gene contains a typical TATA box and numerous potential binding sites for transcription factors including AP1, AP2, AP3, c-Myb, HNF-5, and NF-IL6. As for the set gene, its promoter region does not have any TATA or CCAAT motif and contains a few potential binding sites for transcriptional factors such as c-Myb and gamma-IRE. When these promoter regions were placed upstream of the CAT reporter gene and transfected into a carp CF cell line, the 5'-upstream 1.6-kb DNA fragment of the snf5 gene displayed stronger promoter activity, approximately three-fold higher than that of the 5'-upstream 1.3 kb DNA fragment of the set gene. By transient expression and immunofluorescent staining, we also showed that the pufferfish SNF5 and SET are nuclear proteins, consistent with their postulated roles as transcriptional factors.

Amino Acid Sequence↗
PubMed1998

mcl-1 is an immediate-early gene activated by the granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling pathway and is one component of the GM-CSF viability response.

mcl-1, a bcl-2 family member, was originally identified as an early gene induced during differentiation of ML-1 myeloid leukemia cells. In the present study, we demonstrate that Mcl-1 is tightly regulated by the granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling pathway. Upon deprivation of survival factor from TF-1 myeloid progenitor cells, Mcl-1 levels quickly dropped prior to visible detection of apoptosis of these cells. Upon restimulation of these deprived cells with GM-CSF, the mcl-1 mRNA was immediately induced and its protein product was accordingly resynthesized. Analysis with Ba/F3 cells expressing various truncation mutants of the GM-CSF receptor revealed that the membrane distal region between amino acids 573 and 755 of the receptor beta chain was required for mcl-1 induction. Transient-transfection assays with luciferase reporter genes driven by various regions of the mcl-1 promoter demonstrated that the upstream sequence between -197 and -69 is responsible for cytokine activation of the mcl-1 gene. Overexpression of mcl-1 delayed but did not completely prevent apoptosis of cells triggered by cytokine withdrawal. Its down regulation by antisense constructs overcame, at least partially, the survival activity of GM-CSF and induced the apoptosis of TF-1 cells. Taken together, these results suggest that mcl-1 is an immediate-early gene activated by the cytokine receptor signaling pathway and is one component of the GM-CSF viability response.

Amino Acid Sequence↗
PubMed1998

The role of CD8+ cells and CD11a in the rejection-reaction of the allogeneic microskin grafting.

By using histological tissue sections and immunohistochemical stains, the rejection-reaction of the allogeneic microskin grafting of rabbit-skin wounds was studied. The results revealed that the rejection started early in the days following transplantation. In the second week following transplantation, severe graft rejection with epithelial destruction was noted. In addition, an increase of CD8+ cells and expression of the adhesion molecule-CD11a-were also noticed and persisted throughout the days following transplantation. These findings suggest that the CD8+ cells and CD11a may play a major role in the rejection of allogeneic skin grafting.

Animals↗
PubMed1997

Suppressive effects of destruxin B on hepatitis B virus surface antigen gene expression in human hepatoma cells.

Destruxin B, a cyclodepsipeptide was originally identified as a plant pathogen from the fungus, Alternaria brassicae. We examined the antiviral activity of destruxin B and found that it suppresses the expression of the hepatitis B viral surface antigen (HBsAg) gene in human hepatoma Hep3B cells which carry an integrated viral gene in its chromosome. In contrast, destruxin B shows no cytotoxic effect on the viability of the cells. Furthermore, it can be shown that destruxin B can reversibly suppress HBsAg production by Hep3B cells in a concentration-dependent manner with EC50 of 0.5 microM. Northern blot analysis indicates that the suppression of HBsAg gene expression by destruxin B is mainly at the mRNA level. Destruxin B not only suppresses the endogenously expressed HBsAg in the Hep3B cells but also suppresses the HBsAg produced either from the stable transfected HBV DNA in another human hepatoma HuH-7 cell line which carry no endogenous HBV genome. These results suggest that destruxin B may have future potential for development as a specific anti-HBV drug.

Antiviral Agents↗
PubMed1997

RF heating of implanted spinal fusion stimulator during magnetic resonance imaging.

Radio frequency (RF) heating of an implanted spinal fusion stimulator (SpF) during magnetic resonance imaging (MRI) was studied on a full-size human phantom. Heating during MRI scans (GE Signa 4X, 1.5 T) was measured with RF-transparent fiberoptic sensors. With the implant correctly connected, the maximum temperature rises were less than 2 degrees C during the 26 min that the scans were at maximum RF power. At the tip of a broken stimulator lead (connecting the SpF generator and its electrodes), the maximum temperature rise was 11-14 degrees C. Regular 4-min scans of the spinal cord produced similar temperature rises at the broken tip. After the generator and the leads were removed, heating at the electrode connector tip was less than 1.5 degrees C. The control temperature rises at the same locations, without the stimulator, were less than 0.5 degree C. This study shows that spinal fusion stimulator heating is within the Food and Drug Administration safety guideline of 2 degrees C. However, if a lead wire is broken, it is unsafe during MRI scans. Radiological examinations will be necessary to ensure the integrity of the implant.

Electric Stimulation↗
PubMed1997

Electrochemical treatment of mouse and rat fibrosarcomas with direct current.

Electrochemical treatment (ECT) of cancer utilizes direct current to produce chemical changes in tumors. ECT has been suggested as an effective alternative local cancer therapy. However, a methodology is not established, and mechanisms are not well studied. In vivo studies were conducted to evaluate the effectiveness of ECT on animal tumor models. Radiation-induced fibrosarcomas were implanted subcutaneously in 157 female C3H/HeJ mice. Larger rat fibrosarcomas were implanted on 34 female Fisher 344 rats. When the spheroidal tumors reached 10 mm in the mice, two to five platinum electrodes were inserted into the tumors at various spacings and orientations. Ten rats in a pilot group were treated when their ellipsoidal tumors were about 25 mm long; electrode insertion was similar to the later part of the mouse study, i.e., two at the base and two at the center. A second group of 24 rats was treated with six or seven electrodes when their tumors were about 20 mm long; all electrodes were inserted at the tumor base. Of the 24 rats, 12 of these were treated once, 10 were treated twice. and 2 were treated thrice. All treated tumors showed necrosis and regression for both mice and rats; however, later tumor recurrence reduced long-term survival. When multiple treatments were implemented, the best 3 month mouse tumor cure rate was 59.3%, and the best 6 month rat tumor cure rate was 75.0%. These preliminary results indicate that ECT is effective on the radiation-induced fibrosarcoma (RIF-1) mouse tumor and rat fibrosarcoma. The effectiveness is dependent on electrode placement and dosage.

Animals↗