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C Chang

Publications and source records attributed to C Chang.

At least 361 records · Page 20Linked to original sources

Distribution of androgen and estrogen receptor mRNA-containing cells in the rat brain: an in situ hybridization study.

The distribution of cells that express mRNA encoding the androgen (AR) and estrogen (ER) receptors was examined in adult male and female rats by using in situ hybridization. Specific labeling appeared to be largely, if not entirely, localized to neurons. AR and ER mRNA-containing neurons were widely distributed in the rat brain, with the greatest densities of cells in the hypothalamus, and in regions of the telencephalon that provide strong inputs in the medial preoptic and ventromedial nuclei, each of which is thought to play a key role in mediating the hormonal control of copulatory behavior, as well as in the lateral septal nucleus, the medial and cortical nuclei of the amygdala, the amygdalohippocampal area, and the bed nucleus of the stria terminalis. Heavily labeled ER mRNA-containing cells were found in regions known to be involved in the neural control of gonadotropin release, such as the anteroventral periventricular and the arcuate nuclei, but only a moderate density of labeling for AR mRNA was found over these nuclei. In addition, clearly labeled cells were found in regions with widespread connections throughout the brain, including the lateral hypothalamus, intralaminar thalamic nuclei, and deep layers of the cerebral cortex, suggesting that AR and ER may modulate a wide variety of neural functions. Each part of Ammon's horn contained AR mRNA-containing cells, as did both parts of the subiculum, but ER mRNA appeared to be less abundant in the hippocampal formation. Moreover, AR and ER mRNA-containing cells were also found in olfactory regions of the cortex and in both the main and accessory olfactory bulbs. AR and ER may modulate nonolfactory sensory information as well since labeled cells were found in regions involved in the central relay of somatosensory information, including the mesencephalic nucleus of the trigeminal nerve, the ventral thalamic nuclear group, and the dorsal horn of the spinal cord. Furthermore, heavily labeled AR mRNA-containing cells were found in the vestibular nuclei, the cochlear nuclei, the medial geniculate nucleus, and the nucleus of the lateral lemniscus, which suggests that androgens may alter the central relay of vestibular and auditory information as well. However, of all the regions involved in sensory processing, the heaviest labeling for AR and ER mRNA was found in areas that relay visceral sensory information such as the nucleus of the solitary tract, the area postrema, and the subfornical organ. We did not detect ER mRNA in brainstem somatic motoneurons, but clearly labeled AR mRNA-containing cells were found in motor nuclei associated with the fifth, seventh, tenth, and twelfth cranial nerves. Similarly, spinal motoneurons contained AR but not ER mRNA.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

DNA linkage analysis and studies of the androgen receptor gene in a large kindred with complete androgen insensitivity.

DNA linkage analysis of the X chromosome and studies with cDNA probes specific for the androgen receptor gene were performed on the largest known kindred with the syndrome of complete androgen insensitivity. The affected subjects (XY) have absent binding of dihydrotestosterone to the androgen receptor (the receptor negative form of androgen insensitivity). In this kindred there was maternal transmission of the gene, with all affected males expressing complete genital feminization. Linkage analysis studies were conducted with two DNA probes, DXS1 and PGK1, localized to the Xq11-Xq13 region of the long arm of the X chromosome near the centromere. The results demonstrate linkage to the markers in the order of DXS1-(AR; PGK1), thus localizing the AR gene to an area between Xq11 and Xq13. Three cDNA probes that span various parts of the androgen receptor gene, including the DNA and steroid binding domain, were used to evaluate the androgen receptor gene in normal individuals, carrier mothers, and affected subjects. Identical restriction fragment patterns were found in all three groups studied. Thus the androgen receptor gene was present in affected subjects without detectable DNA polymorphism at the androgen binding domain. Therefore, despite complete absence of binding to the androgen receptor, the defect in the androgen receptor gene in this kindred is not the result of a gene deletion. The results point to a mutation or a small insertion/deletion as the probable cause of the syndrome.

Cell Line↗

Quantitative exploration of the REF52 protein database: cluster analysis reveals the major protein expression profiles in responses to growth regulation, serum stimulation, and viral transformation.

Quantitative protein databases reveal the response of cells to experimental variables, such as exposure to growth factors or transfection with a transforming gene. The nature of the response depends on the type of cell and its internal state at the time of the stimulus. By constructing a protein database to study a given cell line, we can better understand the differentiated state of the cell, the growth regulatory mechanisms it employs, the particular mechanisms it uses to cope with its environment, and the ways these mechanisms may have been compromised through mutation or transformation. The REF52 database is a quantitative database designed to study growth control and transformation in a well-defined family of normal and transformed rat cell lines. The database, which has been described and analyzed elsewhere (J. I. Garrels and B. R. Franza, J. Biol. Chem. 1989, 264, 5283-5298 and J. I. Garrels and B. R. Franza, J. Biol. Chem. 1989, 264, 5299-5312) is further explored here using cluster analysis. This method reveals the most common protein expression profiles for each series of two-dimensional gels without requiring any prior hypothesis or queries on the part of the investigator. This study reveals, for each experiment, large and small clusters of protein expression profiles, most of which have readily apparent biological meaning. For example, large clusters of proteins induced or repressed during growth to confluence have been revealed, and several clusters of transformation-sensitive proteins reveal differential effects of transformation by DNA- and RNA-tumor viruses. This analysis extends our earlier quantitative explorations of the REF52 protein database and helps to show how such a database can be used to provide context and guidance for molecular studies of regulation in a given cell system.

Algorithms↗

Role of neuroexcitation in development of blood-brain barrier and oedematous changes following cerebral ischaemia and traumatic brain injury.

Potential involvement of neuroexcitatory mechanisms was studied in: 1) repetitive forebrain ischaemia in gerbils, 2) global cerebral ischaemia in rats and 3) cryogenic injury to the cerebral cortex in rats and gerbils. Uptake of 45Ca was used as a marker of injury, whereas ultrastructural localization of calcium was assessed with an oxalate-pyroantimonate method. The blood-brain barrier was evaluated with immunostaining for serum albumin. Changes in extracellular glutamate were estimated by microdialysis and an enzymatic cycling assay. Changes in water content were assessed by specific gravity measurements. Repetitive ischaemia of 3 x 5 min carotid occlusions produced a cumulative effect with regard to development of oedema and neuronal injury. This was associated with several-fold increments in glutamate release after repeated insults, whereas there was no apparent correlation with energy metabolism disturbances. Other studies revealed in all models a development of secondary foci distant to the primary impact of ischaemia or cold lesions, which were characterized by calcium accumulation in swollen dendrites, chronic neuronal changes and intraneuronal uptake of serum proteins, all of these changes being potentially compatible with involvement of neuroexcitatory mechanisms.

Animals↗

Raising twin babies and problems in the family.

Out of 296 mothers of twins interviewed about difficulties encountered while rearing twins, 49% mentioned inadequate sleep, 18% financial strain, 39% emotional disturbance, 22% disturbance in the marital relationship, and 43% lack of time to take care of other children. With the growth of twins, the problems gradually decreased, except for emotional disturbance, the most difficult period being the first year. Only 15% reported leaving the babies in the care of relatives during the day time. Overall, 68.2% felt the support from the father, but 39% of them felt the support was useless; 49% felt support from the mother-in-law, but 43% of them regarded the mother-in-law to be of no help; 36% were supported by their own mothers, but 54% mentioned that they had received little help. One must conclude that social support from relatives was inadequate. Public policy should take this fact into account and provide public health nurses and social workers to help the mothers of twins, especially during the first year.

Affective Symptoms↗

Identification of genes expressed in the tobacco shoot apex during the floral transition.

The shoot apex of higher plants contains undifferentiated meristematic cells that serve as the origin of post-embryonic organs. The transition from vegetative to reproductive growth results in the commitment of the apical meristem to floral organ formation. To identify the molecular signals that initiate floral development, we have pursued the isolation of genes that are transcriptionally active in the shoot apex of tobacco during the transition from vegetative to floral growth. The small size of the apex led us to utilize polymerase chain reaction shoot apices. This approach enabled the isolation of the apex-specific and floral apex-specific cDNA clones described in this paper. One clone, A3, detected an equivalent level of transcript in the shoot apex during all developmental stages observed. The second clone, FA2, detected a unique transcript that increased in abundance in the shoot apex during the transition to flowering and showed high levels of expression in developing petals, stamens, and pistils.

Base Sequence↗

Immunohistochemical localization of androgen receptors with mono- and polyclonal antibodies to androgen receptor.

Rat, human, and mouse tissues were stained immunohistochemically using mono- and polyclonal androgen receptor antibodies. Monoclonal antibodies were raised in rats and used to stain human and mouse tissues; polyclonal antibodies were raised in rabbits and used to stain rat tissues. Frozen tissue sections were incubated with the appropriate androgen receptor antibody and staining was completed by the indirect avidin-biotin peroxidase method. A comprehensive survey of rat and mouse tissues was performed. Antibody staining was found exclusively in the nucleus of certain specific cell types, suggesting that the androgen receptor is a nuclear protein. All male sexual organs in the rat showed strong positive nuclear staining for androgen receptor. Weaker positive reactions were seen in kidney, liver, adrenal cortex and pituitary gland. Furthermore, positive staining for androgen receptor was exhibited in skeletal, cardiac and smooth muscle cells, and central nervous tissue. Female reproductive organs also contained androgen receptor-positive cells. The spleen was found to be the only organ examined which did not stain for androgen receptor. The monoclonal antibody could also demonstrate androgen receptor-positive cells in a human prostatic cancer and in a prostate with benign hyperplasia. These data demonstrate the use of antibodies in revealing cellular/subcellular distribution of androgen receptor in target tissues.

Animals↗

Colonoscopic removal of a gallstone obstructing the sigmoid colon.

For the patient with a single gallstone obstructing the colon, this technique offers nonoperative relief of the obstruction, allowing definitive operative therapy to be accomplished at a later time on a more elective basis. The patient should be carefully observed during the interval for intervening recurrent gallstone ileus.

Aged↗

Molecular cloning of new human TR2 receptors: a class of steroid receptor with multiple ligand-binding domains.

Previously we isolated a new group of cDNA clones from human testis cDNA libraries which might code for new steroid receptors. The cDNA and predicted amino acid sequences of two of these receptors, named TR2-5 and TR2-7 receptors, were determined. We report here the nucleotide and deduced amino acid structures of two other receptors that we named TR2-9 and TR2-11 receptors. The calculated MW of TR2-5 receptor, TR2-7 receptor, TR2-9 receptor and TR2-11 receptor are 52,982, 20,528, 50,849 and 67,223 respectively, which match well with the apparent MW of in vitro translated products. The 26 amino acids involved in the formation of "Zn-fingers" are conserved. The ligand-binding domain of TR2-9 receptor is 16 amino acids shorter and has 3 different amino acids compared with TR2-5 receptor. The TR2-11 receptor has a ligand-binding domain which is longer and quite different compared with the other TR2 receptors. The multiple ligand-binding domains of TR2 receptor could be the products of different genes or may be due to RNA splicing errors. So far, we have failed to find binding activity with any known steroid hormone; this promotes the possibility that an unidentified steroid hormone may be involved.

Amino Acid Sequence↗

Identification, characterization, and purification of two mammalian stress proteins present in mitochondria, grp 75, a member of the hsp 70 family and hsp 58, a homolog of the bacterial groEL protein.

We describe the identification, characterization, and purification of two members of the mammalian stress protein family, both of which are shown to be components of the mitochondria. The first, with an apparent mass of 58,000 daltons, is a constitutive protein whose synthesis increases in cells exposed to elevated temperatures and/or amino acid analogs and is therefore referred to as heat shock protein hsp 58 (hsp 58). The second, with an apparent mass of 75,000 daltons, is also a constitutive protein whose synthesis is increased in cells following glucose deprivation or exposure to either a calcium ionophore or 2-deoxyglucose and therefore represents a member of the so-called glucose-regulated proteins (grp 75). In cells treated with the potassium ionophore, nonactin, both hsp 58 and grp 75 were observed to accumulate in precursor form. Since nonactin has been reported to specifically inhibit the processing of cytoplasmic precursor proteins destined for the mitochondria, we investigated whether mature hsp 58 and grp 75 were components of the mitochondria. Mitochondria were isolated from rat liver and shown to contain both hsp 58 and grp 75. Indirect immunofluorescence using antibodies specific to either hsp 58 or grp 75 confirmed their presence within mitochondria. Proteolytic digestion experiments with intact mitochondria indicated that both proteins were not accessible to external proteolytic attack, suggesting that they are not exposed on the cytoplasmic face of the outer membrane. Based on a variety of biochemical and immunological criteria, grp 75 is shown to be a member of the hsp 70 family of stress proteins, while hsp 58 represents the mammalian equivalent of the bacterial groEL protein. Procedures for the purification of both hsp 58 and grp 75 are presented. The possible biochemical role of these two mitochondrial stress proteins is discussed in relation to the known biochemical function of their related stress protein counterparts.

Animals↗

Analysis of gp74 expression by transformed rat fibroblasts from experimental pulmonary metastases following specific ricin A-chain immunotoxin therapy.

Ricin A chain immunotoxin (IT) 45-2D9-RTA mediates regression of spontaneous pulmonary metastases and lung colonies from K-ras transformed rat fibroblasts (TRF cells). However, residual metastases are frequently noted after IT therapy, and therefore, possible mechanisms mediating tumor cell escape were investigated. Individual lung colonies were dissected from lungs of BALB/c mice, and single-cell suspensions of fresh cells from short-term cultures (eight passages) were tested. Immunoperoxidase staining with 45-2D9 monoclonal antibody showed that stable loss of surface antigen by cells cultured from IT-treated mice did not occur after four injections of specific IT. Sensitivity to specific IT in vitro was equal for metastatic tumor cells from mice treated with either two or four doses of specific IT compared to cells from nonspecific IT-treated mice and to parental cells. Clones derived from metastases of IT-treated mice were not resistant to IT. Clones derived from metastases of specific IT-treated mice internalized bound antibody or IT at the same rate as untreated cells. Freshly disaggregated cells from specific IT-treated mice were as sensitive to specific IT as were cells from nonspecific IT-treated or untreated mice. Specific IT successfully mediated reduction of lung colonies derived from fresh suspensions of lung colony TRF cells from IT-treated mice. This reduction was equivalent to that seen for cells not previously exposed to specific IT. Immunoperoxidase stains of lung sections with 45-2D9 showed that colonies consisting entirely of unstained TRF cells were present in both specific IT and phosphate buffered saline-treated mice. There was a trend toward a higher percentage of antigen-negative colonies in mice treated with IT, although 9 days following specific IT therapy, greater than 80% of lung colonies expressed gp74 antigen. When TRF cells were grown on agar plugs, which promoted three-dimensional growth, groups of cells showing absence of immunoperoxidase staining with antibody to gp74 were identified during 2 weeks of growth. Thus, stability of antigen-negative variants is favored by three-dimensional growth conditions and the selective pressure of IT administration. Our results also suggest that impaired trafficking of IT to antigen-positive cells may also contribute to escape from IT therapy.

Animals↗

Variations in lymphokine generation by individual lymph nodes draining human malignant tumors.

Individual lymph nodes draining tumors vary in their degree of immunological activity. Cell suspensions from tumor-free nodes located relatively near to tumors are spontaneously less reactive and respond poorly to exogenous stimulation by mitogens and lymphokines. Diminished spontaneous uptake of tritiated thymidine by lymph node cells not exposed to exogenous stimulation suggests that tumor-proximate immune suppression exists in vivo and is not purely a laboratory artefact. The present study was undertaken to explore that possibility further. Fluid in which cell suspensions from tumor-free nodes were prepared, and supernatants from short-term cultures of nodes located at different distances from tumors were compared for their capacity to inhibit the in vitro migration of the human lymphoblastoid cell line QIMR-WIL. Inhibitory activity of fluids from individual nodes was related to their position relative to the tumor and their immune competence, assessed by the responses to mitogens of cell suspensions prepared from them. Cell suspension fluids from 92/111 nodes (83%) significantly inhibited the migration of QIMR-WIL, at a level similar (44 +/- 14%) to that induced by the supernatants of mixed lymphocyte cultures (43 +/- 17%). Fluids from the nodes of melanoma patients were more inhibitory than those from breast cancer patients (49 +/- 12% and 37 +/- 13%, respectively, P = 0.003). The inhibitory activity of the different nodes of individual node groups varied significantly in 25 of 33 patients (76%), the node nearest the tumor generating least inhibitory activity (indexing the greatest immune suppression) in 20 of these 25 patients (80%). The strength of migration-inhibitory activity was concordant with the responsiveness to mitogen stimulation in up to 14 of 18 patients (78%). Studies of molecular size and heat stability indicated that the inhibitory factors had characteristics consistent with common migration-inhibitory lymphokines such as leukocyte-migration-inhibitory factor, macrophage-inhibitory factor and interleukin-2. Our findings further support the hypothesis that lymph nodes nearest to tumors are relatively immune-suppressed in vivo.

Body Fluids↗

Prostate androgen receptor: immunohistological localization and mRNA characterization.

Four androgen receptor (AR) specific monoclonal antibodies were used for the immunohistochemical localization of AR in the human prostate tissue. The prostate tissue consisted of alveoli embedded in fibromuscular stroma and lined with a single layer of columnar secretory epithelial cells. The immunoreactive ARs were found predominantly in the nuclei of epithelial cell, suggesting ARs, like estrogen receptors and progesterone receptors, are mainly nuclear proteins. Northern blot hybridization showed that AR mRNA is about 9 kilobases (kb) and relative abundant in the androgen-sensitive organs, such as ventral prostate, dorsolateral prostate and seminal vesicle.

Blotting, Northern↗

Isolation and characterization of human TR3 receptor: a member of steroid receptor superfamily.

Complementary DNAs (cDNAs) encoding a member of steroid receptor super-family, named TR3 receptor, were isolated from a human prostate lambda gt11 cDNA library on the basis of homology of oligonucleotide probes to the DNA-binding domain common to members of the steroid receptor super-family. Expression of TR3 receptor cDNA produced a 64 kDa DNA-binding protein in a rabbit reticulocyte lysate. Nucleotide sequence analysis showed that TR3 receptor cDNA contains two regions of sequences which correspond to the DNA- and hormone-binding domains of members of the steroid receptor super-family. The amino acid sequences in the hormone-binding domain of the TR3 receptor shares about 20% homology with estrogen receptor and less than 15% homology with other known steroid receptors. The DNA-binding domain of the TR3 receptor has about 55% homology with all other known steroid receptors. TR3 receptor had 86% nucleotide and 91% amino acid sequence homology with mouse NUR/77, suggesting that TR3 receptor may be a human homologue of mouse NUR/77 gene product.

Amino Acid Sequence↗

Induction of the TRPM-2 gene in cells undergoing programmed death.

RNA and protein products encoded by the testosterone-repressed prostate message-2 gene (TRPM-2) are induced to high levels, coordinate with the onset of cell death, in numerous rodent models of inducible tissue damage. These models include cell death initiated by hormonal stimuli (prostate regression), pressure insult (renal atrophy after ureteral obstruction), developmental stimuli (necrosis of interdigital tissue), and cytotoxic injury (chemotherapeutic regression of a tumor). Sequence analysis of cDNA encoding TRPM-2 revealed its close homology with a product referred to as SGP-2 or clusterin expressed constitutively by Sertoli cells; however, the immunologically related polypeptides expressed in regressing tissues differ in molecular mass from the forms secreted by the testis. Although the function(s) of the products encoded by the TRPM-2 gene remains unclear, their presence provides a remarkable and early indicator of programmed cell death in many types of mammalian cells.

Animals↗

[Prevalence of temporomandibular disorder syndrome (TMD) in university students--a third year report of the epidemiological study in Taiwan].

A total of 2033 university students (1161 boys, 872 girls, ages ranged from 17 to 32 years) were examined according to an examination form. Pain and functional disorders of the head and neck muscles and TM joints, as well as the dental occlusal conditions were recorded. Questionnaires relating to personal history, oral habits, experience of pain and disorder, and psychoemotional status were also given and replied. It was found that 42.9% of the students had one or more signs of TMD, and girls suffered slightly more often. The prevalence was not as high as in junior college students, and was about the same as in teenagers reported in the previous two years. Occlusal factors like slide in centric, balancing side contact, dental restoration and molar guidance during protrusion were more often found in the TMD group, while missing teeth, incisal overlap, attrition and cross bite were not. About 34% of the students had a previous experience of TM joint pain and/or clicking while only 20% of them had such symptoms during the examination. More than 60% of the students having joint clicking were not aware of this symptom. The oral habit and bruxism were not found to be more often in the TMD group. Emotional factors such as Type A personality, emotion, stress, anger, etc. had a higher score in the TMD group while tension, anxiety, and competition did not. It could be concluded that TMD is a fairly common syndrome in the youth of Taiwan although not serious or treatment demanded. Psychoemotional factors are as important as dental factors in relation to the TMD symptomatology, if not more important.

Adolescent↗