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H Chang

Publications and source records attributed to H Chang.

At least 163 records · Page 9Linked to original sources
PubMed1996

4(R)- and 4(S)-(5-phenylpentyl)-glutamic acids attenuate excitatory postsynaptic potential on hippocampus without inhibiting postsynaptic NMDA and AMPA receptors.

The electrophysiological effects of 4(R)- and 4(S)-(5-phenylpentyl)-glutamic acids (C5R and C5S analogues) were investigated. In the presence of 5 microM C5R or C5S, excitatory postsynaptic potentials (EPSPs), elicited by stimulating the Schaffer collateral-commissural pathway, recorded in the hippocampal CA1 region on rat brain slices were reduced by more than 50% of the control. Both the NMDA receptor- and the non-MNDA receptor-mediated components of the EPSP were attenuated to the similar extent. The blockade of the presynaptic input, by 0.5 microM tetrodotoxin (TTX), resulted in that C5 analogues no longer inhibited the depolarization induced by AMPA or NMDA on brain slices. On the other hand, the AMPA- and NMDA-induced whole cell inward current of the dissociated hippocampal neurons was not affected by the presence of C5 analogues, investigated with the whole cell patch-clamping technique. These results suggest that the site of the inhibitory action of C5R and C5S may be at the presynaptic termini and that the release of the excitatory neurotransmitter, most likely the glutamate, may be tampered.

Animals↗
PubMed1995

Angiotensinogen gene expression is induced by cyclical mechanical stretch in cultured rat cardiomyocytes.

The effect of cyclical mechanical stretch on angiotensinogen gene expression was examined using a neonatal rat cardiocyte culture system. Cultured cardiocytes grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation, at 60 cycles/min. The angiotensinogen gene was activated 2 to 5 fold after stretch for 3 to 24 hr, as shown by quantitative reverse transcription polymerase chain reaction. The 5'-flanking region of the angiotensinogen promoter was activated after stretch for 24 hr. This gene expression could be completely suppressed by losartan, a specific antagonist of angiotensin II receptor. These results indicate that (1) cyclical mechanical stretching of cardiocytes is a good model for the study of cardiac hypertrophy-related gene expression; (2) cyclical stretch up-regulates expression of the angiotensinogen gene and (3) the increase in promoter activity may contribute to the induction of angiotensinogen mRNA by cyclical stretch.

Angiotensinogen↗
PubMed1995

Regulation of human cardiac myosin heavy chain genes by cyclical mechanical stretch in cultured cardiocytes.

We examined the effect of cyclical mechanical stretch on the regulation of cardiac myosin heavy chain genes using an isolated neonatal rat cardiocyte culture system. Cultured cardiocytes grown on a flexible membrane were deformed by vacuum to 20% of maximum elongation, at 60 cycles/min in a serum-free medium. Cyclical stretch did not cause myocyte damage as assessed by supernatant LDH measurement and trypan blue exclusion test. The levels of myosin heavy chain (MyHC) mRNA increased as early as 1 h after stretch, reaching 12-fold over the control in 24 h, as shown by Northern blot analysis. However, the proximal 5'-flanking regions of the alpha- and beta-MyHC gene which were linked to chloramphenicol acetyltransferase (CAT) reporter gene did not exhibit enhanced CAT activity following cyclical stretch. Deletion of the chimeric constructs to shorten the 5'-flanking regions of the MyHC genes generated by polymerase chain reaction amplification did not enhance the CAT activity under cyclical stretch. This finding suggests that the stretch-response element of the alpha- and beta-MyHC gene promoter is probably not present in the proximal region of either the alpha- or beta-MyHC genes.

Animals↗
PubMed1995

Cloning and sequencing of the catechol 2,3-dioxygenase gene of Alcaligenes sp. KF711.

The catechol 2,3-dioxygenase is an aromatic ring-fission enzyme catalyzing the conversion of catechol to 2-hydroxymuconic semialdehyde. A catechol 2,3-dioxygenase gene has been cloned from chromosomal DNA of Alcaligenes sp. KF711, and its sequence was determined. The catechol 2,3-dioxygenase gene was consisted of 927 nucleotides with ATG initiation codon and TGA termination codon, which can encode a polypeptide of molecular weight 35 kDa containing 308 amino acid residues. G+C content of the gene was 58 mol%, and a putative ribosome-binding sequence was identified at about 10 nucleotides upstream from the ATG initiation codon. The sequence of catechol 2,3-dioxygenase from Alcaligenes sp. KF711 exhibited 81-92% homology at nucleotide level and 84-92% homology at amino acid level with those of corresponding enzymes encoded in xylE of TOL plasmid, nahH of NAH7 plasmid, and dmpB of Pseudomonas CF600.

Alcaligenes↗
PubMed1995

Reliability of maximum number of simultaneously open channels as an estimator for the number of channels in single-channel recordings.

The single-channel recording technique has revolutionized the electrophysiological study of the ion channels. Interpretation of single-channel recordings often requires the exact number of channels present in a given patch--for example, derivation of the probability of a channel being open--and most theories handling gating kinetics assume that there is only one channel in the patch. Most investigators have estimated the number of channels as the number of maximal simultaneously open channels observed. Although this method is easy to understand and apply, examination of the validity of such estimation has not been much conducted before. As an attempt for such an examination, we have applied computer simulation method in order to obtain practically useful aids in judging the reliability of such estimates. Results show that (i) large estimated values (for example, larger than 10) are likely to be incorrect, and (ii) when the estimated value is small and the derived probability of a channel being open is within a certain range, the estimated number of channels is most likely to be correct.

Animals↗
PubMed1995

Highly efficient adenovirus-mediated gene transfer into renal cells in culture.

In summary, we have shown that adenovirus vector efficiently introduced foreign gene into cultured renal cells both of mesangial and tubular origin. Genes transferred were properly expressed to produce the molecules of expected function. It was possible to introduce the gene into nearly 100% of the cells treated. Expression of the gene began as early as 12 hours after the infection, increased until 48 hours and persisted at least up to eight days. Finally, the vector was non-toxic to the cells, as judged from simple toxicity tests. Successful application of adenovirus vector enables for us to study function of pertinent molecules in suitable host cells and opens a new way for examining renal cellular physiology and pathophysiology.

Adenoviruses, Human↗
PubMed1995

Reduction of reperfusion injury in rat skeletal muscle following administration of cinnamophilin, a novel dual inhibitor of thromboxane synthase and thromboxane A2 receptor.

We used cinnamophilin, a novel dual inhibitor of thromboxane synthase and thromboxane A2 (TXA2) receptor, and superoxide dismutase (SOD) with catalase to examine their protective effect against reperfusion injury in rat skeletal muscle. In 5 groups of 6 wistar rats three hours of ischaemia were induced in one hind limb by application of a tourniquet to the proximal thigh; the contralateral limb served as an internal, nonischaemic control. The first group did not receive any drug nor was it reperfused. In the other four groups, normal saline (reperfusion control), dimethylsulphoxide (DMSO), cinnamophilin, or SOD with catalase was given before removal of the tourniquet and one hour of reperfusion followed. Skeletal muscle injury was measured by a quantitative spectrophotometric assay of triphenyltetrazolium chloride (TTC) reduction and by muscle weight gain. One hour of reperfusion significantly (p<0.05) lowered TTC reduction in ischaemic limbs in the reperfusion control group in comparison with the rats in 3h ischaemia alone. Among the four reperfusion groups, only the cinnamophilin group had significantly lower decrease of TTC reduction and significantly lower muscle weight gain. These results demonstrate the protective effect of cinnamophilin against reperfusion injury of the ischaemic skeletal muscle in rats.

Animals↗
PubMed1995

Sources of anticipatory emotional distress in women receiving chemotherapy for breast cancer.

BACKGROUND: The contribution of classical conditioning processes to patients' distress before chemotherapy infusions (anticipatory distress) was compared to other potential sources of distress (e.g., trait anxiety). We hypothesized that posttreatment distress (putative unconditioned response) would become a stronger predictor of anticipatory distress as patients underwent more treatment infusions (putative conditioning trials). MATERIALS AND METHODS: Fifty women with early stage breast cancer, undergoing standard chemotherapy, completed questionnaires in the clinic prior to each of eight consecutive treatment infusions, as well as telephone interviews to assess side effects following infusions. RESULTS: Consistent with the conditioning hypothesis, posttreatment distress became significantly related to anticipatory distress at the fourth infusion and became the strongest predictor by the sixth. Path analysis indicated that posttreatment distress had a direct influence on anticipatory distress, and that trait anxiety had an indirect influence by influencing apprehension about chemotherapy which, in turn, directly predicted anticipatory distress. CONCLUSIONS: The results of the present study contribute to an emerging view of anticipatory distress as a conditioned response in chemotherapy patients. Results demonstrate that conditioning factors may be one of the strongest predictors of anticipatory distress in the later phases of chemotherapy treatment.

Adult↗
PubMed1995

Activation of CLN1 and CLN2 G1 cyclin gene expression by BCK2.

The Saccharomyces cerevisiae CLN3 protein, a G1 cyclin, positively regulates the expression of CLN1 and CLN2, two additional G1 cyclins whose expression during late G1 is activated, in part, by the transcription factors SWI4 and SWI6. We isolated 12 complementation groups of mutants that require CLN3. The members of one of these complementation groups have mutations in the BCK2 gene. In a wild-type CLN3 genetic background, bck2 mutants have a normal growth rate but have a larger cell size, are more sensitive to alpha-factor, and have a modest defect in the accumulation of CLN1 and CLN2 RNA. In the absence of CLN3, bck2 mutations cause an extremely slow growth rate: the cells accumulate in late G1 with very low levels of CLN1 and CLN2 RNA. The slow growth rate and long G1 delay of bck2 cln3 mutants are cured by heterologous expression of CLN2. Moreover, overexpression of BCK2 induces very high levels of CLN1, CLN2, and HCS26 RNAs. The results suggest that BCK2 and CLN3 provide parallel activation pathways for the expression of CLN1 and CLN2 during late G1.

Cyclins↗
PubMed1995

Diagnostic accuracy of transesophageal echocardiography for detecting patent ductus arteriosus in adolescents and adults.

To compare the accuracy of transesophageal echocardiography (TEE) with that of transthoracic echocardiography (TTE) in the detection of patent ductus arteriosus (PDA) in the adolescent and the adult, 40 patients with PDA and 50 patients with other congenital heart diseases were studied. All echocardiograms were recorded before cardiac catheterization and surgery. The echocardiographic diagnosis of PDA was made by direct visualization of a shunt flow in the duct. A mosaic flow in the pulmonary artery without direct visualization of the duct was considered possible but not definitely diagnostic of PDA. TEE showed greater sensitivity and negative predictive value than TTE (97% vs 42%, and 98% vs 68%, respectively; p < 0.001) in confirming the diagnosis of PDA. The specificity and positive predictive value in establishing the diagnosis of PDA were the same for both techniques. In the subgroup of patients with Eisenmenger's syndrome, the sensitivity of TEE and TTE in confirming diagnosis of PDA was 100% and 12% (p < 0.01), respectively. The sensitivity of monoplane and biplane TEE in the diagnosis of PDA was comparable (95% and 100%, respectively; p = NS). In conclusion, TEE was highly sensitive and specific in detecting PDA in adolescents and adults. It was also highly valuable for detecting the cause of pulmonary hypertension in patients with Eisenmenger's syndrome.

Adolescent↗
PubMed1995

p53 mutations, c-myc and bcl-2 rearrangements in human non-Hodgkin's lymphoma cell lines.

Fourteen Non-Hodgkin's lymphoma cell lines were generated and assessed for the presence of structural p53, c-myc and bcl-2 gene changes. Single or multiple changes were observed in 11 of the lines. Alterations of the p53 gene were most frequent and documented for 10 lines by immunoprecipitation using the antibodies PAb 240 and PAb 1801, sequencing studies and Southern blot analysis. A detailed study was performed in one of the cell lines (OCI-Ly 4) for which material of the original tumor sample was available. Two point mutations identified by sequencing cDNA derived from the cell line were also present in the original tumor specimen. In contrast, DNA prepared from fibroblasts of the same patient did not show the mutations. Six of the 14 lines demonstrated c-myc rearrangements, while bcl-2 changes were observed in 4. The presence of c-myc was associated with shorter survival of this group of patients with aggressive disease. None of the other changes present as single or composite alterations were correlated with clinical outcome measures.

Adult↗
PubMed1994

In situ transcription with Tth DNA polymerase and fluorescent nucleotides.

We and others have described methods to label specific nucleic acid sequences in fixed cells by reverse in situ transcription (IST). They are simple alternatives to the tedious steps of in situ hybridization with labeled probes. We have favored use of thermostable DNA polymerases after heat denaturation of template secondary structure, accompanied by synthesis of cDNA from an annealed primer, but the approach has been limited by the low reverse transcriptase (RT) activity of Taq polymerase and delayed detection methods. We have improved the technique by the use of recombinant Thermus thermophilus (rTth) DNA polymerase and fluorescein-12-dUTP (FIST). Jurkat T lymphocytes were stimulated with ionomycin + phorbol myristate acetate to produce interleukin-2 (IL-2) mRNA in vitro overnight. They were cytospun onto slides and fixed in 70% ethanol + 30% DEPC-treated water, acetone, and air-dried. The slides were placed on a temperature-controlled heating block, and the cell spot was covered with a plastic coverslip. The temperature was raised to 95 degrees C, and 5-10 microliters of modified Perkin-Elmer/Cetus rTth RT reaction mix was injected under the edge of the coverslip. Each 10 microliters of mix in DEPC-water contained 10 mM Tris-HCl, pH 8.3, 90 mM KCl, 1 mM MnCl2, 1 mM dithiothreitol, 10 U placental ribonuclease inhibitor, 0.125 mM dA,C,GTPs, 0.1 mM fluorescein-12-dUTP, 2 U rTth DNA polymerase, and 4 pM 22-mer oligonucleotide primer, which spanned the second intron of IL-2. After 3 min at 95 degrees C, 1 min at 50 degrees C and 10 min at 72 degrees C, the slides were washed in 0.5 x phosphate-buffered saline, pH 7.0, at 42 degrees C, in 70% ethanol, 100% ethanol, and air-dried. The cells were mounted in antifade solution (2% n-propyl gallate in 70% glycerol), and could be viewed immediately by fluorescence microscopy. Image analysis showed that stimulated Jurkat cells were brighter than uninduced controls or those treated with RNase or without polymerase or primer. FIST appears to be useful for the detection of specific mRNAs in single cells.

Base Sequence↗
PubMed1994

Potentiation of progesterone receptor-mediated transcription by the immunosuppressant FK506.

The nontransformed steroid receptors contain several non-steroid binding proteins, such as hsp90, hsp70, and p59. Recently, we and others have shown that p59 (FKBP59) is an immunophilin which binds two potent immunosuppressants, FK506 and rapamycin. This raises the possibility that FK506 or rapamycin may modify the function of steroid receptors. To develop this line of inquiry, we chose a yeast model system in which the human progesterone receptor form B (hPR-B) was cotransformed with a reporter gene. The reporter contains two copies of a progesterone response element/glucocorticoid response element (PRE/GRE) upstream of the CYC1 promoter which are linked to the lacZ gene of Escherichia coli. We found that FK506 potentiated the ability of progesterone in activating transcription. To gain insight into the mechanism of FK506's regulation of PR action, we questioned whether calcineurin is involved, because it has been shown that FK506 is a specific inhibitor of calcineurin, a Ca(2+)- and calmodulin-regulated phosphatase, through the formation of an FKBP12-FK506-calcineurin-calmodulin complex. We found that 15-O-desmethyl-FK520, an FK506 analogue which is an excellent ligand of FKBP12, but a poor inhibitor of calcineurin, failed to induce the same effect as FK506. We also found that calmidazolium, a calmodulin antagonist, mimicked FK506's action. Furthermore, immunoblot analysis showed that both FK506 and calmidazolium potentiated the effect of progesterone in decreasing the mobility of hPR-B upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This suggests that FK506 and calmidazolium may cooperate with progesterone in increasing the level of hPR-B phosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcineurin↗