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Biomedical subjects

C C Chang

Publications and source records attributed to C C Chang.

At least 379 records · Page 21Linked to original sources
PubMed1993

Effects of epidermal growth factor on growth control and signal transduction pathways in different human hepatoma cell lines.

The roles of epidermal growth factor (EGF) on cell growth control and phosphatidylinositol signal transduction pathway in human hepatoma cell lines with different differentiated states were evaluated. Ligand binding study showed that only one receptor type with similar affinity was found in all three cell lines. Under serum-free conditions, EGF (10(-8)-10(-11) M) enhanced DNA synthesis and cell proliferation of the three cell lines in a dose-dependent manner. The response of the poorly differentiated HA22T/VGH hepatoma cells was most obvious whereas much smaller effects were found in Hep3B and Chang liver cells. The metabolism of phosphoinositides also increased in HA22T/VGH cells as compared with both Hep3B and Chang liver cells under basal and EGF-treated conditions. Our data indicated that EGF had different effects on different human hepatoma cell lines and its role might be more important in poorly differentiated hepatoma cells than in well differentiated ones.

Carcinoma, Hepatocellular↗
PubMed1993

Effects of insulin on TGF-beta 1-induced cell growth inhibition in the human hepatoma cell lines.

The effects of insulin on cell growth control by transforming growth factor beta 1 (TGF-beta 1) in human hepatoma cell lines were studied. TGF-beta 1 inhibited cell growth and DNA synthesis of Hep3B cells but not that of HA22T/VGH cells. The cell cycle-dependent p34cdc2 kinase activity was inhibited by TGF-beta 1 in a dose-dependent manner in Hep3B cells. In contrast, the p34cdc2 kinase activity of HA22T/VGH cells was not regulated by TGF-beta 1. When insulin (10(-7) M) was added simultaneously with TGF-beta 1, we found that the inhibitory effects of TGF-beta 1 on cell growth and DNA synthesis in Hep3B cells was completely blocked and the p34cdc2 kinase activity of Hep3B cells was recovered after insulin administration. Thus, cell growth inhibition by TGF-beta 1 in Hep3B hepatoma cells can be antagonized by insulin and their interaction may play an important role in the tumor progression stage of hepatocarcinogenesis.

CDC2 Protein Kinase↗
PubMed1993

Status of tryptophan residue in cobrotoxin and alpha-bungarotoxin.

Acrylamide quenching studies indicated that the exposure degree of the Trp residue in cobrotoxin was higher than that in alpha-bungarotoxin. The Trp residue of cobrotoxin was in a positively charged environment as revealed by iodide quenching, while the Trp residue of alpha-bungarotoxin was not accessible for iodide. Analysis of hydrophilicity profile and local concentration of positively charged residues of toxin molecule also indicated that Trp in cobrotoxin was in a highly hydrophilic and positively charged environment. Measurement of Trp fluorescence with increasing temperature showed that the stability of environment of Trp in alpha-bungarotoxin was higher than in cobrotoxin. Result of competitive binding for nicotinic acetylcholine receptor (AchR) between cobrotoxin and alpha-bungarotoxin revealed that the molecular interaction of the two toxins with AchR was not the same. These, together with the fact that the cationic groups of the two toxins are involved in the binding with AchR, suggest that the observed different environment surrounded Trp residue and different AchR binding mechanism might fulfill a different requirement of the invariant Trp in the lethality of cobrotoxin and alpha-bungarotoxin.

Animals↗
PubMed1992

Chemical modification of arginine residues in alpha-bungarotoxin.

The reaction of alpha-bungarotoxin (alpha-BuTX) with 1,2-cyclohexanedione resulted in the modification of only Arg-72 but arginine at position 36 or 72, as well as both were modified by reaction of the toxin with p-hydroxyphenylglyoxal. No derivative modified at Arg-25 was obtained, indicating that this residue may be located in the interior region of alpha-BuTX molecule. Monoderivative at Arg-72 showed about 50% of the lethal toxicity and binding activity of alpha-BuTX to nicotinic acetylcholine receptor (AChR), while the activity was decreased to one-third when the invariant Arg-36 was modified, indicating that the latter residue is more closely related to the interaction of the toxin with AChR. Approx. 13% of the residual activity was observed when both arginine residues at 36 and 72 were modified. The antigenicity of alpha-BuTX was still retained essentially intact after Arg-36 or -72 was modified, whereas it decreased to 50% when both these arginine residues were modified. The present study indicates that Arg-36 and -72 in alpha-BuTX may be involved in the multipoint contact between the toxin and AChR, but neither is absolutely essential for the binding.

Amino Acids↗
PubMed1992

Transcriptional activation of human (2'-5')oligoadenylate synthetase gene expression by the phorbol ester 12-O-tetradecanoyl-phorbol 13-acetate in type-I-interferon-treated HL-60 and HeLa cells.

(2'-5')Oligoadenylate [(2'-5')(A)n] synthetase is a key enzyme in the interferon-elicited antiviral response whose controlled expression in interferon-treated cells has been only partially elucidated. In this investigation, we have compared the modulation of the (2'-5')(A)n synthetase gene by interferon alone and by the combination of interferon and a second cellular effector, 12-O-tetradecanoyl-phorbol 13-acetate (TPA). Although TPA alone had no effect on (2'-5')(A)n synthetase, it potentiated the induction of (2'-5')(A)n of synthetase by interferon in HL-60 and HeLa cells by increasing content of its mRNA and an immunoreactive 40-kDa isoenzyme. Since TPA activates protein kinase C (PKC), other PKC-activating phorbol-ester analogues were tested and found to be effective, whereas the PKC inhibitor staurosporine reduced the potentiative activity of TPA. By using the (2'-5')(A)n synthetase gene promoter linked to a reporter gene, chloramphenicol acetyltransferase (CAT), TPA and interferon were found to result in a doubling of CAT activity compared to cells treated with interferon alone. Moreover, when nuclear extracts prepared from control cells or cells treated with TPA and interferon (IFN), separately or together, were incubated with radioactively labeled oligodeoxynucleotides containing the interferon-responsive element (IRE), TPA was shown to down-regulate an IFN-inducible IRE/protein complex. These data further suggest that TPA regulates (2'-5')(A)n synthetase gene expression at the level of transcription.

2',5'-Oligoadenylate Synthetase↗
PubMed1992

Antagonism by beta-eudesmol of neostigmine-induced neuromuscular failure in mouse diaphragms.

beta-Eudesmol, a sesquiterpenol extracted from a Chinese herb, Atractylodes lancea, at 10-80 microM, did not affect muscle action potentials, miniature and evoked endplate potentials and acetylcholine-induced depolarization in the presence or absence of neostigmine in mouse phrenic nerve-diaphragms. However, the tetanic fade, muscle fasciculation and twitch potentiation induced by neostigmine were effectively antagonized by 20 microM beta-eudesmol. When trains of pulses were applied to the nerve in the presence of neostigmine, beta-eudesmol reduced the incidence of explosive depolarization of the endplate from 95% to 35-67% of junctions, and shortened the duration when it occurred. Moreover, both the maximal and steady-state depolarizations during repetitive stimulation were reduced while the amplitudes of steady-state endplate potentials were increased. The results suggest that beta-eudesmol antagonized neostigmine-induced neuromuscular failure mainly by a presynaptic action to depress the regenerative release of acetylcholine during repetitive stimulation. The mechanism of antagonism is obviously not tubocurarine-like and it is unrelated to desensitization of acetylcholine channels.

Acetylcholine↗
PubMed1992

Microbial adherence on poly(methyl methacrylate) (PMMA) surfaces.

Infection remains a major complication following the use of implanted biomaterials. Often these infections are caused by low-virulence organisms or by a mixture of organisms (polymicrobial). In this study two methods were used to quantitate the bacteria which had adhered to poly(methyl methacrylate) (PMMA) samples. The bacteria were eluted from the sample using an ultrasonic cleaner. The number eluted was then counted by colony counts, which determines viable organisms and by particle counting which counts both viable and nonviable organisms. A known adherent strain of Staphylococcus epidermidis and a strain of Proteus mirabilis were used. In general the adherence of S. epidermidis was greater than that of Proteus. When the two organisms were used together, there was an alteration in the adherence pattern which generally increased the adherence of Proteus and had no effect or decreased the adherence of S. epidermidis. The use of both quantitation techniques provided important information on the adherence of organisms to PMMA to which gentamicin had been added. It was evident that organisms did adhere to the PMMA plus gentamicin samples but were not viable when eluted. The amount of adherence to PMMA plus gentamicin was similar to that of PMMA alone at 30 min but was markedly decreased at 24 h. There was a considerable dead biofilm mass on the PMMA plus gentamicin samples which might be a significant promoter of late infections by providing a surface attractive to other strains of bacteria.

Bacterial Adhesion↗
PubMed1992

Growth of MCF-7 human breast carcinoma in severe combined immunodeficient mice: growth suppression by recombinant interleukin-2 treatment and role of lymphokine-activated killer cells.

The severe combined immunodeficient (SCID) mouse, lacking functional T and B lymphocytes, has been considered by many groups to be a prime candidate for the reconstitution of a human immune system in a laboratory animal. In addition, this immuno-deficient animal would appear to have excellent potential as a host for transplanted human cancers, thus providing an exceptional opportunity for the study of interactions between the human immune system and human cancer in a laboratory animal. However, because this animal model is very recent, few studies have been reported documenting the capability of these mice to accept human cancers, and whether or not the residual immune cells in these mice (e.g. natural killer, NK, cells; macrophages) possess antitumor activities toward human cancers. Thus, the purpose of this study was (a) to determine whether or not a human breast carcinoma cell line (MCF-7) can be successfully transplanted to SCID mice, (b) to determine whether or not chronic treatment of SCID mice with a potent lymphokine (recombinant interleukin-2, rIL-2) could alter MCF-7 carcinoma growth, and (c) to assess whether or not rIL-2-activated NK cells (LAK cells) are important modulators of growth of MCF-7 cells in SCID mice. To fulfill these objectives, female SCID mice were implanted s.c. with MCF-7 cells (5 x 10(6) cells/mouse) at 6 weeks of age. Six weeks later, some of the mice were injected i.p. twice weekly with rIL-2 (1 x 10(4) U mouse-1 injection-1). Results clearly show that MCF-7 cells can grow progressively in SCID mice; 100% of the SCID mice implanted with MCF-7 cells developed palpable measurable tumors within 5-6 weeks after tumor cell inoculation. In addition, MCF-7 tumor growth was significantly (P less than 0.01) suppressed by rIL-2 treatment. rIL-2 treatment was non-toxic and no effect of treatment on body weight gains was observed. For non-tumor-bearing SCID mice, splenocytes treated in vitro with rIL-2 (lymphokine-activated killer, LAK, cells) or splenocytes derived from rIL-2-treated SCID mice (LAK cells) had significant (P less than 0.01) cytolytic activity toward MCF-7 carcinoma cells in vitro. In contrast, splenocytes (LAK cells) derived from tumor(MCF-7)-bearing rIL-2-treated SCID mice lacked cytolytic activities toward MCF-7 cells in vitro. No significant concentration of LAK cells in MCF-7 human breast carcinomas ws observed nor did rIL-2 treatment significantly alter growth of MCF-7 cells in vitro.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Sequence determination and characterization of a phospholipase A2 isozyme from Trimeresurus gramineus (green habu snake) venom.

In addition to phospholipase A2-I (PLA2-I) reported previously (ODA et al., 1991, Toxicon 29, 157), a new PLA2 named PLA2-II was isolated from Trimeresurus gramineus (green habu snake) venom, and its amino acid sequence was determined by sequencing the native protein and the peptides produced by enzymatic (Achromobacter protease I and clostripain) cleavages of the carboxamidomethylated derivative of the protein. The protein consisted of 122 amino acid residues and His-47, Asp-48, and Asp-98 which have been assumed to be essential for PLA2 activity were conserved. Its sequence similarity to PLA2-I was 79%, with 26 residual differences. In contrast to the unique presence of Phe-28 in PLA2-I, PLA2-II contains Tyr-28 as seen in most of other PLA2s. There was no significant difference between the dissociation constants of PLA2-I and PLA2-II for Ca2+. Secondary structure compositions of PLA2-II were similar to those of PLA2-I and Crotalus atrox PLA2. A striking difference was found between these isozymes in contractile activity of isolated smooth muscle preparation of guinea-pig ileum. PLA2-II was over ten times more potent than PLA2-I, although its lipolytic activity toward egg-yolk was even slightly weaker (73%) than that of PLA2-I. The difference in contractile activities of PLA2-I and PLA2-II could be assumed to be due to discriminative lipid recognition brought about by different amino acid residues at the 58th position (Asp for PLA2-I and Asn for PLA2-II).

Alcaligenes↗
PubMed1992

G protein mediated signal transduction: membrane immunoglobulin associated phosphoproteins identified in octyl-beta-glucoside lysates of normal B cells.

Several recent studies have shown that the antigen receptors (mIgM and mIgD) on normal B cell surfaces are part of larger multimolecular complexes, as each is noncovalently coupled to at least two other proteins whose phosphorylation is inducible upon G protein activation. However, B cell receptor complexes appear to be extremely labile, so that extraction of membrane immunoglobulin (mIg) from cell lysates in physical association with noncovalently coupled proteins has been shown to be highly dependent on the choice of detergent. Recent progress has to a large extent been dependent upon the introduction of digitonin, a detergent in which B cell receptor complexes are at least partially stable, as an agent to solubilize B cell membranes. Studies presented here demonstrate that if another well known detergent, octyl-beta-D-glucopyranoside (octyl glucoside), is utilized in place of digitonin, mIg receptors are immunoprecipitated from B cell lysates in association with several newly identified proteins, whose phosphorylation is also G protein dependent. In particular we find three to four mIg associated phosphoproteins in the molecular weight range of 47 to 57 kDa, and at least two others at 82 and 112 kDa. Some of these proteins may correspond to polypeptides that have been functionally linked to mIg dependent signal transduction, but have not as yet been shown to be physically coupled to the receptor.

Animals↗
PubMed1992

Inhibition of Moloney murine leukemia virus reverse transcriptase activity by tetrahydroxyxanthones isolated from the Chinese herb, Tripterospermum lanceolatum (Hyata).

Five tetrahydroxyxanthones (THXs) isolated from Tripterospermum lanceolatum (Hyata) have been shown to have a strong inhibitory effect on Moloney murine leukemia virus reverse transcriptase (Mo-MLV RT) activity when (rA)n-(dT)15 and (rC)n-(dT)12-18 were used as template-primers. 50% inhibitory concentrations of 1,3,5,6-THX, 2,3,6,7-THX 1,3,6,7-THX, 3,4,5,6-THX, and 3,4,6,7-THX were determined to be 0.15, 0.27, 0.58, 0.12, and 0.12 microM, respectively. Their effects were concentration-dependent, and the mode of inhibition was found to be by competitive inhibition with respect to template-primer. The tetrahydroxyl groups of THXs were shown to be important for their inhibitory activity. Acylation of THXs with various groups resulted in a moderate or strong decrease in their inhibitory activity.

Acylation↗
PubMed1992

Perspectives on formaldehyde toxicity: separating fact from fantasy.

Formaldehyde (IUPAC name, methanal) is one of the simplest, most ubiquitous molecules in our environment and troposphere. Exposure to large amounts of formaldehyde can produce a variety of respiratory and dermatologic problems in humans, in both the home and the workplace. However, in spite of anecdotal reports on formaldehyde-induced illness over the past 20 years there is a paucity of data regarding its potential as either an allergen or an antigen in humans. In addition, many of our current impressions about formaldehyde are based on studies of dubious scientific validity. In this review, we discuss the biological and chemical properties of formaldehyde and its presence in materials which we come in contact with, and finally attempt to put in perspective our current understanding of the detrimental effects of formaldehyde on our health, or lack thereof. There is no evidence at present that formaldehyde causes immunological diseases. Finally, and unfortunately, many of the studies have drawn invalid conclusions and are based on poorly controlled anecdotal observations.

Animals↗
PubMed1992

Inhibition by neosurugatoxin and omega-conotoxin of acetylcholine release and muscle and neuronal nicotinic receptors in mouse neuromuscular junction.

Neosurugatoxin and omega-conotoxin, known to be specific ligands for the neuronal nicotinic receptor and Ca2+ channel, respectively, were previously claimed to exert no depressant action on the mouse neuromuscular junction. It was found that in preparations partially blocked with tubocurarine or with low Ca(2+)-high Mg2+ Tyrode's, both toxins, at 3-10 microM, depressed indirect twitches and either produced wanings (neosurugatoxin) or waxings (omega-conotoxin) of indirectly elicited tetanic contractions whilst in normal Tyrode's the contractile forces were not changed. In normal Tyrode's, neosurugatoxin decreased the amplitudes of spontaneous and evoked endplate potentials and enhanced the run-down of endplate potentials as did tubocurarine though with lesser potency. By contrast, omega-conotoxin (10 microM) decreased the amplitude of the evoked but not of the spontaneous endplate potential in low Ca(2+)-high Mg2+ Tyrode's, and produced facilitation of endplate potentials, instead of run-down, on repetitive stimulations. Higher concentrations of omega-conotoxin appeared to depress quantal release in normal Tyrode's. The effects were all reversible. The prolonged endplate depolarization found in preparations treated with neostigmine or 3,4-diaminopyridine, was partially depressed by both toxins. The results suggest that neosurugatoxin blocks the neuron and muscle nicotinic receptors in the neuromuscular junction with comparable potency. The pharmacology of the nicotinic receptor on motor nerve terminal seems more similar to the muscle nicotinic receptor than to that on autonomic ganglia or brain. On the other hand, omega-conotoxin seems to block a small fraction of Ca2+ channels on the motor nerve and decreases the quantal release of evoked endplate potentials.

Acetylcholine↗
PubMed1992

Cloning of a creatinase gene from Pseudomonas putida in Escherichia coli by using an indicator plate.

A genomic library of Pseudomonas putida DNA was constructed by using plasmid pBR322. Transformants of Escherichia coli in combination with Proteus mirabilis cells grown on creatinase test plates were screened for creatinase activity; transformants were considered positive for creatinase activity if a red-pink zone appeared around the colonies. One creatinase-positive clone was further analyzed, and the gene was reduced to a 2.7-kb DNA fragment. A unique protein band (with a molecular weight of approximately 50,000) was observed in recombinant E. coli by minicell analysis.

Cloning, Molecular↗
PubMed1992

Spurious hyperchloremia and decreased anion gap in a patient with dextromethorphan bromide.

Although cold syrup containing dextromethorpan bromide is widely administered, the bromism due to cold syrup has not been reported. We report a patient who had negative anion gap with hyperchloremia and conscious loss because of daily intake of cold complex syrup (containing dextromethorphan bromide 0.4 mg/ml, acetaminophen 8.33 mg/ml) for headache for 4-5 years. The bromide content in cold complex syrup resulted in serum levels of bromide that interfered with the automated analyzers for chloride content. When conscious change is due to bromism, hemodialysis instead of forced hydration and diuresis should be performed immediately. Therefore, patients with a markedly negative anion gap with hyperchloremia should be considered as having halide intoxication.

Bromides↗
PubMed1992

Changes of mitochondrial respiratory functions and superoxide dismutase activity during liver regeneration.

The general objective of this study was to examine the relationship between mitochondrial respiratory function and liver regeneration in the rat. The role that free radicals may play in the process was also evaluated. It was found that the respiratory control and ADP/O ratios were concomitantly decreased to the lowest level at 6 hr after hepatectomy and gradually recovered thereafter. Both ratios were significantly increased at 48 hr and quickly reached plateau levels. Assays of mitochondrial respiratory functions revealed that the activities of Complex I+III, Complex II+III and Complex IV all decreased drastically at 6 hr after hepatectomy and then gradually returned to the original level during 18-24 hr after hepatectomy. Interestingly, the activities of all these enzyme complexes continuously increased and were elevated significantly above the normal levels (145-200%). In contrast, the liver mitochondrial electron transport activities of sham-operated rats returned only to the original level after recovering from the operation-induced decline at 6 hr post-hepatectomy. We measured the superoxide dismutase (SOD) activity of liver mitochondria of the hepatectomized and sham-operated rats. The results showed that the Mn-SOD activity started to increase after hepatectomy, reached a maximum (900% of control) at 6 hr, and then returned to normal levels at 24 hr after operation. The Cu, Zn-SOD activity was increased 9-fold in hepatectomized rats and about 3-fold in sham-operated rats as compared with control rats. The maximum activity of Mn-SOD was found to be about 4 times higher than that of Cu, Zn-SOD after hepatectomy. The amount of lipoperoxides in the liver mitochondria was found to be increased to 140% in hepatectomized rats and to 120% in sham-operated rats as compared with that of the control rats. Taken together these results suggest that the changes in mitochondrial respiratory functions in the early phase of hepatectomy are due to tissue damage caused by the transient elevation of free radicals. These free radicals are then quickly disposed of by the ever-increasing activities of the Mn-SOD and Cu, Zn-SOD in the liver mitochondria, thereby protecting the liver from further damage and gearing the organ to the regeneration process.

Animals↗