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J Cui

Publications and source records attributed to J Cui.

At least 181 records · Page 10Linked to original sources
PubMed1994

Gating of IsK expressed in Xenopus oocytes depends on the amount of mRNA injected.

IsK is a K+ channel of the delayed rectifier type widely distributed throughout both excitable and nonexcitable cells. Its structure is different from other cloned K+ channels and molecular details of its gating remain obscure. Here we show that the activation kinetics of IsK expressed in Xenopus oocytes depend upon the amount of its mRNA injected, with larger amounts resulting in slower activation kinetics with a longer initial delay during activation. Similar changes in activation kinetics occur with time after a single injection of IsK mRNA. We present two kinetic schemes which illustrate how our experimental results could arise. Both imply an interaction among individual channel proteins during IsK activation. The dependence of channel gating on mRNA concentration provides a novel mechanism for long term regulation of ion current kinetics.

Animals↗
PubMed1994

Effect of CL-316,243, a thermogenic beta 3-agonist, on energy balance and brown and white adipose tissues in rats.

The objective was to assess the effect of a new, highly selective beta 3-adrenergic agonist, CL-316,243 (CL) (J. D. Bloom, M. D. Dutia, B. D. Johnson, A. Wissner, M. G. Burns, E. E. Largis, J. A. Dolan, and T. H. Claus., J. Med. Chem. 35: 3081, 1992), on energy balance and brown and white adipose tissues (BAT and WAT, respectively) in young rats eating a high-fat diet to induce obesity. Chronic treatment with CL increased body temperature and 24-h energy expenditure, mainly by increasing resting metabolic rate. Food intake was not altered but carcass fat was reduced. Interscapular BAT was markedly hypertrophied, with three- to fourfold increases in the content of uncoupling protein (UCP) and cytochrome oxidase. Quantitative immunoelectron microscopy of interscapular BAT of CL-treated rats showed smaller mitochondria with an unchanged total amount of UCP per mitochondrion. The relative frequency of the four major cell types in BAT (mature brown adipocytes, preadipocytes, interstitial cells, endothelial cells) was not altered. The CL-induced hypertrophy differed from that induced by chronic stimulation by endogenous norepinephrine (as in cold-adaptation) in absence of hyperplasia (there was a slightly reduced DNA content), absence of an increase in the thyroxine (T4) 5'-deiodinase activity, and absence of a selective increase in UCP concentration. WAT depots weighed less and had fewer cells (lower DNA content) in the CL-treated rats. Some multilocular adipocytes appeared in these normally almost exclusively unilocular WAT depots (mesenteric, inguinal, epididymal, retroperitoneal). We conclude that CL not only promotes BAT mitochondrial proliferation and thermogenesis and overall energy expenditure and leanness, but also retards the development of WAT hyperplasia during the early stage of diet-induced obesity.

Adipose Tissue↗
PubMed1994

Studies on atrophic change of soleus muscle and its countermeasures in suspended rat.

In the environment of microgravity, the disused atrophy of skeletal muscle, especially leg's muscle, would occur. The three purposes of this study were: 1. To observe the dynamic changes of disused atrophy of skeletal muscle under simulated weightlessness; 2. To approach the mechanism of disused atrophy of muscle; 3. To approach the countermeasures for reducing the degree of atrophy of muscle.

Animals↗
PubMed1993

ATPase activity of TyrR, a transcriptional regulatory protein for sigma 70 RNA polymerase.

The TyrR protein of Escherichia coli is the chief transcriptional regulator of several genes essential for aromatic amino acid biosynthesis and transport. It was established in previous studies that this protein binds ATP, that the TyrR.ATP complex has enhanced affinity for tyrosine, and that the susceptibility of the TyrR protein to hydrolysis by trypsin is altered by ATP. Here we show that the TyrR protein has ATPase activity, which is stimulated by tyrosine. In this respect the TyrR protein resembles the transcriptional activator NtrC. The NtrC protein contains an internal polypeptide segment, 220 amino acid residues in length, with a high degree of identity to the TyrR protein, that contains the presumptive ATPase catalytic center.

Adenosine Triphosphatases↗
PubMed1993

The TyrR protein of Escherichia coli, analysis by limited proteolysis of domain structure and ligand-mediated conformational changes.

The TyrR protein of Escherichia coli K12 is a homodimer containing 513 amino acids/subunit. This protein is important in the transcriptional regulation of several genes whose protein products catalyze steps in aromatic amino acid biosynthesis or transport. Methods were developed for efficiently purifying the TyrR protein to apparent homogeneity. We analyzed the pattern of cleavage of the TyrR protein by trypsin, either in the absence of ligands or in the presence of saturating levels of L-tyrosine, ATP, or poly(dI-dC). At low (1:200 ratio by weight) trypsin levels, in the absence of ligands, two major digestion products accumulated. These were polypeptides of 22 and 31 kDa, shown to contain amino acid residues 1-190 and 191-467, respectively. The pattern of trypsin cleavage was unaffected by tyrosine. In the presence of ATP, an intermediate species of 53 kDa, probably containing amino acid residues 1-467, was observed. The kinetics of appearance of the 53-kDa species were consistent with a role for ATP in accelerating the hydrolysis of the R467-F468 peptide bond. The 53-kDa polypeptide underwent further tryptic hydrolysis to yield fragments of 22 and 31 kDa. When both tyrosine and ATP were present, the rate of formation of the 22- and 31-kDa fragments was more rapid than in the absence of these ligands. It appears that when both ligands are bound, the rates of hydrolysis of peptide bonds R190-Q191 and R467-F468 are both enhanced. Additional limited proteolysis experiments suggested that polypeptide segment 191-467 contains ATP binding site(s), and that the rate of cleavage of peptide bonds R190-Q191 and R467-F468 is altered when the TyrR protein interacts with poly(dI-dC), an analog of target DNA. Our results reveal the presence of two major structural domains within the TyrR protein. The first domain (amino acid residues 1-190) is extremely resistant to hydrolysis by trypsin. The second domain (residues 191-467), which is likely to contain ATP-binding site(s), is homologous to several other transcriptional activators specific for promoters responsive to the sigma 54 form of RNA polymerase. The remainder of the TyrR protein (residues 468-513) contains the operator recognition elements, probably arranged in the form of a helix-turn-helix motif. This polypeptide segment was not detected as a discrete tryptic hydrolysis product.

Adenosine Triphosphate↗
PubMed1993

The change of melanocyte cytotoxicity after systemic steroid treatment in vitiligo patients.

Although there is evidence of a strong association between antimelanocyte autoantibodies and vitiligo, the etiology of vitiligo is still unclear. To elucidate the biological significance of antimelanocyte autoantibodies in vitiligo, we measured the percent cytotoxicity of melanocyte mediated by autoantibody and complement in normal controls (n = 31) and in the patients with active, progressive vitiligo (n = 37). Significant differences in percent cytotoxicity of melanocyte were seen between the control and vitiligo groups (P = 0.0001). There were no significant differences in the percent cytotoxicity of melanocyte between the patients (n = 24) with more than 1 year duration and those (n = 13) with less than 1 year duration. The change of percent cytotoxicity between pre- and post-treatment groups (n = 29) with systemic steroid showed significant differences (P = 0.0243). These findings support the hypothesis that a decrease in the antibody-mediated cytotoxicity against melanocytes may play a role in the improvement of vitiliginous lesions after systemic steroid treatment.

Adolescent↗
PubMed1993

Cytolytic antibodies to melanocytes in vitiligo.

Patients with vitiligo have been found to have circulating antibodies to pigment cells. To evaluate the functional activity of these antibodies, a highly sensitive europium release assay was used to compare complement-mediated cytolysis of human melanocytes by sera of 56 patients with vitiligo (20 with active disease, 25 with inactive disease, 11 with unidentified disease activity) and 47 control individuals. Significant melanocyte lysis was mediated by 32 (57%) of the patients with vitiligo but by only three (6%) of the control sera (p < 0.001), and by 17 (85%) of 20 patients with active vitiligo versus 11 (44%) of 25 patients with inactive disease (p < 0.025). Mean melanocyte lysis by vitiligo sera was 24% versus 6% by control sera (p < 0.0001). A subset of 12 vitiligo sera with high titers of cytolytic antibodies to melanocytes (34% mean cytolysis) reacted minimally (< 2% mean cytolysis) to a panel of control cells that included human and murine melanomas, human fibroblasts, lung carcinoma, and rhabdomyosarcoma. These findings indicate that antibodies present in patients with vitiligo have the functional ability to selectively kill melanocytes and are more common in active disease. These observations support, but do not prove, the hypothesis that vitiligo is an autoimmune disease and that anti-pigment cell antibodies have a role in inducing the disease.

Antibodies↗
PubMed1993

Mutational uncoupling of the transcriptional activation function of the TyrR protein of Escherichia coli K-12 from the repression function.

The tyrosine repressor (TyrR) protein of Escherichia coli can function either as a transcriptional enhancer or as a repressor. The structural basis for these opposite effects was analyzed in specific tyrR deletion mutants constructed in vitro. The functional behavior of the mutant TyrR proteins was evaluated in vivo by using single-copy lacZ reporter systems based on the mtr promoter (10-fold activation by wild-type TyrR protein, mediated by phenylalanine or tyrosine) or the aroF promoter (over 20-fold repression by wild-type TyrR protein, mediated by tyrosine). A mutant TyrR protein lacking amino acids 2 to 9 was completely devoid of transcriptional activation function. Five additional mutant TyrR proteins lacking progressively greater numbers of N-terminal amino acids were likewise activation defective. The mutant TyrR proteins lacking amino acid residues 2 to 9 or 2 to 19 were essentially identical to the wild-type TyrR protein in their ability to repress the aroF promoter. Three other TyrR mutant proteins, lacking up to 143 amino acid residues from the N-terminal end of the protein, retained the ability to repress the aroF promoter, to different extents, in a tyrosine-dependent manner.

DNA Mutational Analysis↗
PubMed1993

A mutational analysis of the structural basis for transcriptional activation and monomer-monomer interaction in the TyrR system of Escherichia coli K-12.

In response to the binding of tyrosine or phenylalanine, the TyrR protein (513 amino acids) activates certain promoters and represses others. In a previous study (J. Cui and R. L. Somerville, J. Bacteriol. 175:303-306, 1993), it was shown that promoter activation was selectively abolished in mutant proteins lacking amino acid residues 2 to 9. An additional series of constructs that encoded mutant TyrR proteins having deletions or point mutations near the N terminus were analyzed. Residues Arg-2 and Leu-3 were shown to be critical for the activation of the mtr promoter. In confirmation of previous findings, none of the activation-defective mutant TyrR proteins had lost significant repression function. The TyrR protein was shown by chemical cross-linking to be dimeric. The polypeptide segments critical for dimer formation in vivo were identified by evaluating the negative dominance phenotypes of a series of mutant proteins, all defective in DNA binding, lacking progressively greater numbers of amino acid residues from either the N terminus or the C terminus. Amino acid residues 194 to 438 were found to contain all of the essential dimerization determinants.

Amino Acid Sequence↗
PubMed1993

[Studies on the pleomorphism of trachoma inclusion].

A series of trachoma inclusion color photograph is taken from the conjunctival smears of 48 trachoma cases. These smears were Giemsa-stained and stored in the slide-boxes for more than 30 years with no distinct changes in color. A description is given to the Pleomorphism of trachoma inclusion and its relation to the host cells. According to the gradual morphological changes of Chlamydia trachomatis multiplied in the infected cells, a 5-staged classification for trachoma inclusions is provided which may be benefit to understanding the pleomorphism of the trachoma inclusion and clinical laboratory diagnosis.

Chlamydia trachomatis↗
PubMed1993

Follow-up of asymptomatic myocardial ischemia in patients with diabetes mellitus.

Early detection of asymptomatic myocardial ischemia in patients with diabetes mellitus (DM) is very important for the prevention of painless myocardial infarction. The purpose of this study is to explore the clinical significance of asymptomatic transient ST changes in ECG in patients with DM and to further clarify the relation between asymptomatic myocardial ischemia and myocardial infarction in DM.

Aged↗
PubMed1992

An improved europium release assay for complement-mediated cytolysis.

An improved assay for complement-mediated cytolysis is described. The target cells are labeled with europium complexed to diethylenetriaminopentaacetate (Eu-DTPA). Cytolysis caused by antibody plus complement leads to the release of the Eu-DTPA complex which is then formed into a highly fluorescent chelate by the addition of 2-naphthoyltrifluoroacetone (2-NTA). The amount of europium chelate formed--a measurement of cell death--is then quantified with a time-resolved fluorometer. The results of the assay are reproducible. Complement-mediated cytolysis when measured by europium release was five times more sensitive than when measured by conventional 51Cr release and three times than when measured by trypan blue exclusion. Because europium does not decay, target cells can be labelled in batches and stored frozen until use, which speeds and simplifies the assay. Thus, europium release assay is a simple and quantitative method to measure complement-mediated cytolysis which is sensitive and more rapid than conventional assays.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1992

Identification of pigment cell antigens defined by vitiligo antibodies.

Patients with vitiligo have circulating antibodies to pigment cells. To characterize this response further and to identify the antigens defined by vitiligo antibodies, sera of 23 patients with vitiligo and 22 patients with unrelated conditions were analyzed by immunoprecipitation and SDS-PAGE analysis of 125I-labeled cell antigens on pigment and control cells. Antibodies to pigment cell antigens were present in 18 (78%) of the patients with vitiligo but in only three (14%) of the control patients (p less than 0.05). The antibodies were directed to one or more antigens with molecular weight (MW) in kilodaltons (kD) of approximately 35, 40-45, 75, 90, or 150. The responses were most commonly directed to the 40-45-kD, 75-kD, and 90-kD antigens. Antibodies to these antigens were present in 74%, 57%, and 35% of vitiligo patients versus in 14%, 9%, and 0% of control individuals. The 35-kD and 90-kD antigens were preferentially expressed on human pigment cells, whereas the 40-45-, 75-, and 150-kD antigens were expressed on both pigment and control cells. These antigens were labeled by the lactoperoxidase technique, suggesting that they are cell surface antigens. These results confirm that antibodies to pigment cells are associated with vitiligo. These antibodies are directed to several cell surface antigens, some of which are preferentially expressed on pigment cells.

Antibodies↗
PubMed1992

Expression and characterization of a canine hippocampal inwardly rectifying K+ current in Xenopus oocytes.

1. An inwardly rectifying potassium current expressed in Xenopus laevis oocytes injected with canine hippocampal poly(A)+ RNA was investigated with the two-microelectrode voltage clamp technique. 2. Xenopus oocytes injected with canine hippocampal poly(A)+ RNA expressed a current activated by hyperpolarization. This current contained an instantaneous and a time-dependent component. Both components were inwardly rectifying and could be blocked by extracellular Cs+ or Ba2+. 3. The expressed current was carried mainly by K+. Its reversal potential measured in different [K+]os could be fitted by the Nernst equation with a slope of -50.7 per tenfold change in [K+]o. Extracellular Cl- and Na+ made minimal contributions to the current. 4. The activation of the expressed current depended on both voltage and [K+]o. Activation started near EK and the activation curve shifted along the voltage axis in parallel with EK when [K+]o was altered. 5. The activation time constants of the expressed current also depended on both voltage and [K+]o. The voltage dependence of the time constants was bell-shaped and the peak value was at a potential 30-50 mV more negative than EK. The voltage dependence of the time constants shifted along the voltage axis when EK was changed. 6. The poly(A)+ RNA extracted from canine hippocampus was fractionated in a 10-31% linear sucrose gradient. The size of the mRNA required to express the inwardly rectifying current was estimated to be around 4 kb. 7. In conclusion, the expressed current is an inwardly rectifying potassium current. The canine hippocampal mRNA should be an excellent source for expression-cloning of the inward rectifier channel.

Animals↗
PubMed1992

Rapid but transient atrophy of brown adipose tissue in capsaicin-desensitized rats.

Our previous studies showed atrophy of brown adipose tissue (BAT) in capsaicin-desensitized rats during the period 11-28 days after injections [Cui et al., Am. J. Physiol. 259 (Regulatory Integrative Comp. Physiol. 28): R324-R332, 1990]. The objective of the present studies was to assess the rapidity with which the atrophy occurred and the extent to which recovery had occurred by 8 wk. Rats, either vehicle-injected controls or capsaicin injected, were studied 1, 3, 14, 28, and 52 days after the last injection. BAT was markedly atrophied at 1 day, having less total protein, fewer mitochondria (less total cytochrome oxidase and total uncoupling protein), and fewer cells (less DNA). Atrophy persisted for up to 14 days but had largely disappeared by 28-52 days. A transient reduction in body weight gain and white epididymal adipose tissue weight had also reversed by 28-52 days. We suggest that the rapid atrophy of BAT after capsaicin desensitization is secondary to the loss of sensory neuropeptides in its sensory nerves, neuropeptides that either exert a trophic effect on synthesis of mitochondria or an inhibitory influence on processes that promote degradation of mitochondria. The retardation of the normal age-associated increase in DNA content of BAT in the capsaicin-desensitized rat suggests that sensory neuropeptides might also modulate cell proliferation.

Acclimatization↗