PubMed Health⌕ Search

Biomedical subjects

C Chang

Publications and source records attributed to C Chang.

At least 217 records · Page 12Linked to original sources
PubMed1997

Functional interaction of the carboxylic acid group of agonists and the arginine residue of the seventh transmembrane domain of prostaglandin E receptor EP3 subtype.

Prostaglandin (PG) E2 binds to PGE receptor EP3 subtype and induces Gi activity. To assess the role of the interaction of the carboxylic acid group of agonists and its putative binding site, Arg-309 in the seventh transmembrane domain of EP3alpha receptor, in receptor activation, we have mutated the positively charged Arg-309 to the polar but uncharged Gln (EP3alpha-R309Q) and Asn (EP3alpha-R309N), and to the non-polar Leu (EP3alpha-R309L). Wild-type, EP3alpha-R309Q and EP3alpha-R309N receptors showed high-affinity binding for PGE2, but the EP3alpha-R309L receptor showed very-low-affinity binding. Guanosine 5'-[gamma-thio]triphosphate increased the PGE2 binding to the wild-type receptor, decreased the binding to EP3alpha-R309Q and EP3alpha-R309N receptors, but did not affect that to the EP3alpha-R309L receptor. Furthermore we examined the Gi activities of two types of EP3 agonist, TEI-3356 with a negatively charged carboxylic acid, and TEI-4343, a methyl ester of TEI-3356 with an uncharged but polar group, towards those receptors. Both agonists inhibited the forskolin-stimulated cAMP formation in wild-type, EP3alpha-R309Q and EP3alpha-R309N receptors in the same concentration-dependent manner, but the agonists showed a very low inhibition of EP3alpha-R309L receptor. These findings demonstrate that the hydrogen-bonding interaction of EP3 agonists and residue 309 in the seventh transmembrane domain of the EP3alpha receptor is sufficient for the functional activation of the EP3alpha receptor.

Animals↗
PubMed1997

Human fatty aldehyde dehydrogenase gene (ALDH10): organization and tissue-dependent expression.

Mutations in the fatty aldehyde dehydrogenase gene (ALDH10) are responsible for Sjögren-Larsson syndrome (De Laurenzi et al., 1996). In this study, the expression and the genomic organization of the ALDH10 gene are reported. The gene spans approximately 31 kb and consists of 10 exons and 9 introns. All exon-intron junction sequences match the classical GT/AG rule. Both S1 nuclease protection assay and primer extension study suggest that the transcription initiation site is located 195 nucleotides upstream from the ATG codon. No canonical TATA box can be found in the 5'-flanking sequence of the gene, but a CCAAT-like box was found 58 bp upstream of the putative transcription start site. Sequence analysis of the 5'-flanking region revealed numerous potential binding sites for transcription factors Sp1 and AP-2 and one putative HIP-1 binding site. Northern blot analysis of poly(A)+ RNA from various tissues revealed two mRNA species, with sizes around 4.0 and 2.0 kb, that are derived from the differential use of two polyadenylation sites. Although this gene is expressed in a variety of human tissues, the expression level of ALDH10 in the liver and skeletal muscle appears to be higher than that in other tissues examined.

Aldehyde Oxidoreductases↗
PubMed1997

Induction of the intronic enhancer of the human ciliary neurotrophic factor receptor (CNTFRalpha) gene by the TR4 orphan receptor. A member of steroid receptor superfamily.

A conserved hormone response element, CNTFR-DR1 (5'-AGGTCAGAGGTCAGG-3'), has been identified in the 5th intron of the alpha component of the ciliary neurotrophic factor receptor (CNTFRalpha) gene for the human TR4 orphan receptor (TR4). Electrophoretic mobility shift assay showed a specific binding with high affinity (Kd = 0.066 nM) between TR4 and the CNTFR-DR1. A reporter gene assay using chloramphenicol acetyltransferase demonstrated that the 5th intron of CNTFRalpha has an enhancer activity which could be induced by TR4 in a dose-dependent manner. Furthermore, our in situ hybridization data showed that abundant TR4 transcripts were detected in adult brain, in regions of cortical and hippocampal neurons, as well as in many developing neural structures, including brain, spinal cord, ganglia (sympathetic and sensory), and neuronal epithelia (retinal, otic, olfactory, and gustatory). The striking similarities in the expression patterns of TR4 and CNTFRalpha in the developing and postnatal nervous systems further support the potential role of TR4 in neurogenesis. Collectively, these data suggest that the human CNTFRalpha gene could represent the first identified neural-specific gene induced by TR4.

Adult↗
PubMed1997

cDNA sequence analysis and expression of kappa-bungarotoxin from Taiwan banded krait.

The cDNAs encoding kappa-bungarotoxin was constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus by reverse transcription-polymerase chain reaction. A high degree of nucleotide sequence homology was observed between kappa-bungarotoxin and other kappa-neurotoxins. The kappa-bungarotoxin was subcloned into the expression vector pET32a(+) and transformed into BL21(DE3) E. coli strain. The recombinant toxin was expressed as a fusion protein. Recombinant kappa-bungarotoxin was separated from the fused protein by cleavage with CNBr and purified by reversed phase high performance liquid chromatography. In addition to kappa-bungarotoxin, the cDNA fragment encoding kappa3-bungarotoxin was also found in the cDNA mixtures prepared from the cellular RNA of the venom glands of the same snake. This result suggests that the venom glands of Taiwanese B. multicinctus should secrete at least two kinds of kappa-neurotoxins.

Amino Acid Sequence↗
PubMed1997

Cloning and expression of the gene for xylose isomerase from Thermus flavus AT62 in Escherichia coli.

The gene encoding xylose isomerase (xylA) was cloned from Thermus flavus AT62 and the DNA sequence was determined. The xylA gene encodes the enzyme xylose isomerase (XI or xylA) consisting of 387 amino acids (calculated Mr of 44,941). Also, there was a partial xylulose kinase gene that was 4 bp overlapped in the end of XI gene. The XI gene was stably expressed in E. coli under the control of tac promoter. XI produced in E. coli was simply purified by heat treatment at 90 degrees C for 10 min and column chromatography of DEAE-Sephacel. The Mr of the purified enzyme was estimated to be 45 kDa on SDS-polyacrylamide gel electrophoresis. However, Mr of the cloned XI was 185 kDa on native condition, indicating that the XI consists of homomeric tetramer. The enzyme has an optimum temperature at 90 degrees C. Thermostability tests revealed that half life at 85 degrees C was 2 mo and 2 h at 95 degrees C. The optimum pH is around 7.0, close to where by-product formation is minimal. The isomerization yield of the cloned XI was about 55% from glucose, indicating that the yield is higher than those of reported enzymes. The K(m) values for various sugar substrates were calculated as 106 mM for glucose. Divalent cations such as Mn2+, Co2+, and Mg2+ are required for the enzyme activity and 100 mM EDTA completely inhibited the enzyme activity.

Aldose-Ketose Isomerases↗
PubMed1997

Metabolic alterations produced by 3-nitropropionic acid in rat striata and cultured astrocytes: quantitative in vitro 1H nuclear magnetic resonance spectroscopy and biochemical characterization.

Quantitative high resolution in vitro 1H nuclear magnetic resonance spectroscopy was employed to study the metabolic effects of 3-nitropropionic acid associated with aging from perchloric acid extracts of rat striata. Systemic injection of 3-nitropropionic acid in rats at a dose of 10 mg/kg/day for seven consecutive days significantly impaired energy metabolism in rats one, four and eight months of age, as evidenced by a marked elevation of succinate and lactate levels. However, a significant decrease in N-acetyl-L-aspartate level, a neuronal marker, was observed in four- and eight-month-old rats but not in one-month-old rats. This would indicate that rats at four to eight months are more susceptible to 3-nitropropionic acid than those at one month. A significant decrease in GABA level was observed in four-month-old 3-nitropropionic acid-treated rats, which is consistent with the literature that GABAergic neurons are particularly vulnerable to 3-nitropropionic acid treatment. In addition, glutamine and glutamate levels were markedly decreased at four and eight months in 3-nitropropionic acid-treated rats. Since glutamine is synthesized predominantly in glia, the observation above suggests that 3-nitropropionic acid intoxication may involve perturbation of energy metabolism, glial injury and consequent neuronal damage. Astrocytes which are essential in the metabolism of glutamate and glutamine were used to further assess 3-nitropropionic acid-induced toxicity. Glial proliferation, mitochondrial metabolism and glutamine synthetase activity were all reduced by 3-nitropropionic acid treatment with a concomitant increase, in a dose-dependent manner, of lactate levels, suggesting that 3-nitropropionic acid is also detrimental to astrocytes in vivo and thus may affect metabolic interaction between neurons and glia. These results not only imply that 3-nitropropionic acid blocks energy metabolism prior to exerting neurotoxic damage but also demonstrate that the degree of energy depletion determines the detrimental effects of 3-nitropropionic acid. In the present study, we also demonstrate that glutamate and glutamine levels as well as astrocytic functions may play pivotal roles in 3-nitropropionic acid-induced striatal lesions.

Animals↗
PubMed1997

LAIR-1, a novel inhibitory receptor expressed on human mononuclear leukocytes.

In the present study, we describe a novel inhibitory receptor, leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1), that is constitutively expressed on the majority of human peripheral blood mononuclear leukocytes. LAIR-1 is a 32 kDa transmembrane glycoprotein with a single immunoglobulin-like domain and a cytoplasmic tail containing two immune receptor tyrosine-based inhibitory motifs. LAIR-1 recruits SHP-1 and SHP-2 phosphatases upon activation, and cross-linking of the LAIR-1 antigen on natural killer (NK) cells results in strong inhibition of NK cell-mediated cytotoxicity. Although it is structurally related to human killer cell inhibitory receptors, LAIR-1 does not appear to recognize human leukocyte antigen (HLA) class I molecules and thus represents a novel HLA class I-independent mechanism of NK cell regulation.

Amino Acid Sequence↗
PubMed1997

When three is less than two: early developments in children's understanding of fractional quantities.

Four experiments examined young children's understanding of the inverse relation between the number of parts into which a quantity is to be divided and the size of each part. In Experiment 1 5-, 6-, and 7-year-old children tended to judge, incorrectly, that bigger shares would result from sharing with more, rather than fewer, recipients. In Experiment 2, 5-year-olds correctly recognized the inverse effect of additional recipients when the sharing was based on subtraction rather than on equal partitioning. In Experiment 3, a modification of the equal-sharing task from Experiment 1 designed to reduce cognitive complexity successfully elicited correct performance from 7-year-olds but not from 5-year-olds. However, 5-year-olds markedly improved when they were given a chance to compare the outcomes of sharing with different numbers of recipients. Experiment 4 corroborated and extended this evidence of learning.

Age Factors↗
PubMed1997

A dominant mutant receptor from Arabidopsis confers ethylene insensitivity in heterologous plants.

Ethylene (C2H4) is a gaseous hormone that affects many aspects of plant growth and development. Ethylene perception requires specific receptors and a signal transduction pathway to coordinate downstream responses. The etr1-1 gene of Arabidopsis encodes a mutated receptor that confers dominant ethylene insensitivity. Evidence is presented here that etr1-1 also causes significant delays in fruit ripening, flower sensecence; and flower abscission when expressed in tomato and petunia plants. The ability of etr1-1 to function in heterologous plants suggests that this pathway of hormone recognition and response is highly conserved and can be manipulated.

Amino Acid Sequence↗
PubMed1997

Localization of androgen receptor in nitric oxide synthase- and vasoactive intestinal peptide-containing neurons of the major pelvic ganglion innervating the rat penis.

Neurons of the rat major pelvic ganglia provide innervation to the pelvic organs and external genitalia. In these ganglia, a subpopulation of neurons containing either nitric oxide synthase or vasoactive intestinal peptide or both molecules, is involved in penile erection. The androgen dependence of penile erection is a well established fact. After castration, decreased testosterone levels have been documented to produce erectile dysfunction possibly resulting from functional alterations in major pelvic ganglion neurons. It was therefore of interest to investigate the presence of androgen receptor within these ganglionic neurons. By using immunohistochemistry and a retrograde labeling technique we have demonstrated that the androgen receptor is present in about 40% of neurons of the major pelvic ganglion innervating the corpora cavernosa of the rat penis. In the major pelvic ganglion, 87% and 81% of the neurons labeled with the fluorescent dye Fast Blue from the penis contained nitric oxide synthase-like immunoreactivity and vasoactive intestinal peptide-like immunoreactivity, respectively. Androgen receptor was present in 20% of neurons containing vasoactive intestinal peptide-like immunoreactivity and about 40% of those containing nitric oxide synthase-like immunoreactivity. These results suggest that androgens, which are known to modulate penile erection, may regulate nitric oxide synthase and vasoactive intestinal peptide within the major pelvic ganglion via a direct interaction with ganglionic neurons.

Amidines↗
PubMed1997

Developmental regulation of the gibberellin biosynthetic gene GA1 in Arabidopsis thaliana.

The GA1 gene of Arabidopsis thaliana encodes ent-kaurene synthase A (KSA), which catalyzes the first committed step in the biosynthetic pathway of the plant hormone gibberellin (GA). Its location in the GA biosynthetic pathway has led to speculation that KSA regulation is one of the controlling steps. However, because KSA activity is so low that it is only measurable in Arabidopsis siliques, GA1 promoter-GUS reporter gene fusions and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were used to examine the expression pattern of GA1. The results from this study indicate that GA1 gene expression is highly regulated during growth and development, and it is restricted to specific cell types at the sites of expression. GA1 promoter activity is highest in rapidly growing tissues, e.g. shoot apicas, root tips, developing flowers and seeds. It is also active in the vascular tissue of some non-growing organs, such as expanded leaves, suggesting that these leaves may be a site of GA synthesis for transport to other organs. It was also found that the first one or two introns in the GA1 gene are required for proper expression. Because of the high degree of regulation, GA1 may act as a gatekeeper, controlling the flow of metabolites into the GA biosynthetic pathway, while the levels of specific bioactive GAs are controlled by other downstream steps.

Alkyl and Aryl Transferases↗
PubMed1997

Mutations associated with Sjögren-Larsson syndrome.

Sjögren-Larsson syndrome (SLS), a rare autosomal disorder characterized by ichthyosis, spastic neurological disorders and oligophrenia, is associated with deficiency of fatty aldehyde dehydrogenase encoded by a gene on chromosome 17q11.2. Mutations of the gene (GDB symbol ALDH10) were recently identified in three SLS patients. Another aldehyde dehydrogenase isozyme. ALDH3, also has a high activity for fatty aldehyde oxidation, and is encoded by a gene in chromosome 17q11.2. Abnormality of the ALDH3 gene could also cause a similar syndrome. The examination of the ALDH3 locus of three additional SLS patients showed that two are heterozygous with C-->G at nt 985 (Pro-->Ala at protein position 329). However, the mutation was found to be common (frequency of the atypical allele is about 0.25) in normal subjects, and not related to SLS. Isoelectric focusing analysis indicated that ALDH3 is hardly expressed in normal as well as patients' fibroblast cells, while ALDH10 expressed in the normal cells is diminished in the three patients' cells. The level of ALDH10 mRNA is also low in the patients' cells. The examination of the ALDH10 locus revealed the existence of a 3 base deletion coupled with a 21 base insertion at intron 6/exon 7 junction in one patient. This abnormality is the same as that found in the patient previously reported by other investigators. One patient is associated with a 2 base deletion at nt 1297 and consequent premature chain termination at protein position 434. Another patient is a compound heterozygote for the same 2 base deletion at nt 1297 and a 5 base insertion at nt 1311 and premature chain termination at protein position 457. Unique characteristics of the SLS mutations are pointed out.

Aldehyde Dehydrogenase↗
PubMed1997

Serum opens tight junctions and reduces ZO-1 protein in retinal epithelial cells.

We have shown previously that serum inhibits tight junction formation in a retinal epithelial cell culture model for the blood-brain barrier. We have now examined in detail the effects of serum on the tight junctions. Our data show that serum induces a breakdown in tight junction function as indicated by decreased transepithelial electrical resistance and increased permeability. Rat serum had effects similar to those of bovine serum, indicating that the activity is species-independent. The effect is concentration-dependent, reversible, and specific for the apical surface, suggesting the involvement of a specific receptor-ligand interaction. Differences in the time course, response magnitude, and structural manifestations between the serum-induced breakdown and that induced by switching the cultures to a low-calcium medium suggest fundamental differences in their mechanisms. The calcium switch results in an immediate and complete junctional breakdown with cell retraction and perinuclear translocation of both actin and the tight junction protein zonula occludens-1. The serum-induced breakdown occurs slowly, is incomplete, and is manifested structurally by decreases in zonula occludens-1 protein, whereas actin organization is unchanged. Thus, serum induces a specific breakdown in retinal epithelial cell tight junctions that may be mediated by effects on the expression of zonula occludens-1.

Animals↗
PubMed1997

Inhibition of hair growth by testosterone in the presence of dermal papilla cells from the frontal bald scalp of the postpubertal stumptailed macaque.

Hair-follicle regression in the bald scalps of stumptailed macaques develops after puberty, which corresponds to an elevation of serum testosterone and dihydrotestosterone. Using the cultured cells from the pre- and postpubertal macaques, we examined the role of dermal papilla cells in testosterone-induced inhibition of outer root sheath cell proliferation. Testosterone showed no effects on proliferation of either dermal papilla cells or outer root sheath cells cultured alone. Testosterone-induced inhibition of outer root sheath cell proliferation occurred only in coculture with dermal papilla cells derived from the bald scalps of adult macaques but not with dermal papilla cells from the hairy occipital scalps of adult macaques or the prebald frontal scalps of juvenile macaques. Furthermore, RU 58841, an androgen receptor blocker, antagonized this testosterone-elicited inhibition. Together our data indicate that the inhibitory effect of testosterone on proliferation of epithelial cells is age dependent, and androgen may play an essential role in hair growth either by inducing repressor(s) from dermal papilla cells, which may then inhibit the growth of epithelial cells of the hair follicle, or by inducing growth factor(s) from dermal papilla cells, which, in turn, may trigger the induction of some repressors in epithelial cells, thereby inhibiting the epithelial cell growth. Our animal studies also showed that RU 58841 has a dramatic effect on hair regrowth in the bald frontal scalp of the stumptailed macaque, which may further support our in vitro culture studies showing that antiandrogens can antagonize testosterone-elicited hair growth. In summary, our studies may provide a model for further isolation of androgen-regulated repressor(s)/growth factors, which may help control hair growth and baldness.

Alopecia↗
PubMed1997

Transcriptional up-regulation of the human androgen receptor by androgen in bone cells.

Androgen regulation of androgen receptor (AR) expression has been observed in a variety of tissues, generally as inhibition, and is thought to attenuate cellular responses to androgen. AR is expressed in osteoblasts, the bone-forming cell, suggesting direct actions of androgens on bone. Here we characterized the effect of androgen exposure on AR gene expression in human osteoblastic SaOS-2 and U-2 OS cells. Treatment of osteoblastic cells with the nonaromatizable androgen 5alpha-dihydrotestosterone increased AR steady state messenger RNA levels in a time- and dose-dependent fashion. Reporter assays with 2.3 kilobases of the proximal 5'-flanking region of the human AR promoter linked to the chloramphenicol acetyltransferase gene in transfected cultures showed that up-regulation of AR promoter activity by androgen was time and dose dependent. Treatment with other steroid hormones, including progesterone, 17beta-estradiol, and dexamethasone, was without effect. The antiandrogen hydroxyflutamide completely antagonized androgen up-regulation. Thus, in contrast to many other androgen target tissues, androgen exposure increases steady state AR messenger RNA levels in osteoblasts. This regulation occurs at least partially at the level of transcription, is mediated by the 5'-promoter region of the AR gene, and is dependent on functional AR. These results suggest that physiological concentrations of androgens have significant effects on AR expression in skeletal tissue.

Bone Neoplasms↗
PubMed1997

A Xenopus type I activin receptor mediates mesodermal but not neural specification during embryogenesis.

Activins and other ligands in the TGFbeta superfamily signal through a heteromeric complex of receptors. Disruption of signaling by a truncated type II activin receptor, XActRIIB (previously called XAR1), blocks mesoderm induction and promotes neuralization in Xenopus embryos. We report the cloning and characterization of a type I activin receptor, XALK4. Like truncated XActRIIB, a truncated mutant (tXALK4) blocks mesoderm formation both in vitro and in vivo; moreover, an active form of the receptor induces mesoderm in a ligand-independent manner. Unlike truncated XActRIIB, however, tXALK4 does not induce neural tissue. This difference is explained by the finding that tXALK4 does not block BMP4-mediated epidermal specification, while truncated XActRIIB inhibits all BMP4 responses in embryonic explants. Thus, the type I and type II activin receptors are involved in overlapping but distinct sets of embryonic signaling events.

Activin Receptors↗
PubMed1997

The distance to community medical care and the likelihood of hospitalization: is closer always better?

OBJECTIVES: This study examined the influence that distance from residence to the nearest hospital had on the likelihood of hospitalization and mortality. METHODS: Hospitalizations were studied for Maine. New Hampshire, and Vermont during 1989 (adults) and for 1985 through 1989 (children) and for mortality (1989) in Medicare enrollees. RESULTS: After other known predictors of hospitalization (age, sex, bed supply, median household income, rural residence, academic medical center, and presence of nursing home patients) were controlled for, the adjusted rate ratio of medical hospitalization for residents living more than 30 minutes away was 0.85 (95% confidence interval [CI] = 0.82, 0.88) for adults and 0.78 (95% CI = 0.74, 0.81) for children, compared with those living in a zip code with a hospital. Similar effects were seen for the four most common diagnosis-related groups for both adults and children. The likelihood of hospitalization for conditions usually requiring hospitalization and for mortality in the elderly did not differ by distance. CONCLUSIONS: Distance to the hospital exerts an important influence on hospitalization rates that is unlikely to be explained by illness rates.

Adolescent↗