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Biomedical subjects

J Zhao

Publications and source records attributed to J Zhao.

At least 649 records · Page 36Linked to original sources
PubMed1994

Isolation and characterization of the reaction product of 4-diazobenzenesulfonic acid and gamma-carboxyglutamic acid: modification of the assay for measurement of beta-carboxyaspartic acid.

gamma-Carboxyglutamic acid (Gla)-containing proteins are extracellular proteins with enhanced cation or mineral-binding properties. Discovery of new Gla-containing proteins is facilitated by methods that decrease the number of steps and time involved in assaying for Gla. Reaction of 4-diazobenzenesulfonic acid (DBS) and Gla or Gla-containing proteins produces an intensely red-colored product. This method has been used to identify Gla-containing proteins in crude extracts of proteins. The reaction product of Gla and DBS has been purified by reversed-phase HPLC and characterized by uv-visible spectroscopy and electrospray ionization (ESI) mass spectrometry (MS). The red-colored product exhibits an absorption maximum at 530 nm. The ESI-MS data of the colored derivative of Gla are consistent with replacement of the two gamma-carboxyl groups in Gla with two DBS groups. DBS also reacts with another malonic acid derivative, beta-carboxyaspartic acid (Asa). The optimum conditions for colorimetric assay of Asa were established, and the potential of this reaction as an assay for Asa and Asa-containing proteins was studied.

1-Carboxyglutamic Acid↗
PubMed1994

Inhibition of DNA replication in cell-free extracts of Xenopus laevis eggs by extracts of Xenopus laevis oocytes.

Cell-free extracts of Xenopus laevis eggs support replication of Xenopus sperm nuclei and purified double-stranded plasmid DNA templates. In contrast, cell-free extracts of Xenopus oocytes do not support replication of these templates. In this study, we show that extracts prepared from Xenopus oocytes, when added to extracts of Xenopus eggs, inhibited replication of Xenopus sperm nuclei and double-stranded plasmid DNA templates. This inhibition did not result from degradation of template DNA by oocyte extracts, nor from inhibition of chain elongation. Nuclei formed in egg extracts in the presence of oocyte extracts were much smaller compared with nuclei formed in egg extracts. Extracts prepared from Xenopus germinal vesicles (oocyte nuclei) also inhibited replication of Xenopus sperm nuclei, but not synthesis of the complementary strand of single-stranded DNA templates in egg extracts. In contrast to extracts prepared from immature oocytes, extracts prepared from Xenopus oocytes matured in vitro effect much less, if any, inhibition of DNA replication. Taken together, these results suggest that molecules that negatively regulate chromosomal DNA replication may be present in Xenopus immature oocytes. The level of these factors may decrease during oocyte maturation, concurrent with the breakdown of oocyte germinal vesicles and manifestation of the capacity of matured oocytes to initiate DNA replication. To our knowledge, this is the first biochemical identification of negative factors that may be involved in the regulation of eukaryotic chromosomal DNA replication.

Animals↗
PubMed1994

Reorganization and condensation of chromatin in mitotic prophase nuclei of Allium cepa.

This paper studies the process and features of chromosome construction in mitotic prophase cells of Allium cepa. The results showed that a prominent reorganization of chromatin occurred during G2--early prophase. The 250-400 nm thick compact chromatin threads in G2 nuclei began to disorganize into about 30, 100 and 220 nm chromatin fibres which constituted the loosely organized chromosome outlines in early prophase before chromosome condensation. In middle prophase, chromosome condensation was characterized by the formation of many condensed regions (aggregates of chromatin), which increased in size (1-1.5 microns) when prophase proceeded. Meanwhile, the chromatin threads that constituted and connected the condensed regions became increasingly thicker (120-250 nm). In late prophase adjacent condensed regions fused to form cylinder-shaped chromosomes. Based on these observations, we come to the conclusion that the construction of prophase chromosomes is a two-step process, that is, the reorganization and condensation of chromatin. In addition, we report the study of silver-stained, DNA- and histone-depleted prophase chromosomes, describe morphological features of the non-histone protein (NHP) residue in early, middle and late prophase chromosomes, and discuss the roles of NHPs in chromosome construction.

Allium↗
PubMed1994

The biological activity of 23-oxa-, 23-oxa-24-oxo-, and 23-thia-dihydroxyvitamin D3.

Three analogs of 1 alpha,25-(OH)2D3 with an oxygen or another heteroatom at position 23 were synthesized in search of separating the cell-differentiating from the calcemic effects of the vitamin D hormone. Their ability to induce superoxide production in human myeloid leukemia cells (HL-60) was 1 alpha,25-(OH)2D3 > 23-oxa-24-oxo-1 alpha,25-(OH)2D3 > 23-thia-1 alpha,25-(OH)2D3 > 23-oxa-1 alpha, 25-(OH)2D3. 23-oxa-24-oxo-1 alpha, 25(OH)2D3 was slightly more potent than 1 alpha,25-(OH)2D3 in inhibiting cell proliferation in MCF-7 cells and 23-thia- and 23-oxa-1 alpha,25(OH)2D3 were less potent. Their in vitro potency to produce osteocalcin in MG-63 cells was 1 alpha,25-(OH)2D3 > 23-oxa-24-oxo-1 alpha,25-(OH)2D3 > 23-thia-1 alpha,25-(OH)2D3 = 23-oxa-1 alpha,25-(OH)2D3. All three analogs had reduced receptor and DBP affinity compared to 1 alpha,25-(OH)2D3. When these analogs were injected in rachitic chicks, only little calcemic effects were observed. The introduction of a heteroatom in carbon 23 of 1 alpha,25-(OH)2D3 thus creates analogs with dissociated action on cell differentiation and calcium homeostasis.

Animals↗
PubMed1994

Polymerase chain reaction kinetics when using a positive internal control target to quantitatively detect cytomegalovirus target sequences.

High-performance liquid chromatography (HPLC) was used to detect and quantify cytomegalovirus (CMV) specific polymerase chain reaction (PCR) products generated during PCR co-amplification. PCR of CMV AD 169 or a plasmid which contains the CMV AD 169 native target sequence using the CMV primer set of Hsia et al. (J. Clin. Microbiol. 27, 1802-1809) generates a 152 bp PCR product. A CMV control sequence plasmid which shared the primer sequence of native CMV AD 169 but when amplified produces a larger 362 bp product was constructed. Under co-amplification conditions there was a linear relationship (over 3 logs) between the molar ratio of input CMV native and control target sequence and the molar ratio of the output PCR products as detected by HPLC despite differences between the two PCR target and product sizes. Co-amplifying known amounts of CMV control sequence plasmid as an internal standard allowed accurate quantitation of the amount of CMV native target sequence in a sample when the two PCR targets were present in approximately equimolar amounts +/- 1.5 log (coefficient of variation (CV) < 12%). By modifying the amount of CMV control target sequence plasmid used for co-amplification, accurate detection of the amount of CMV native sequence in samples could be extended to 5 logs, standard error (S.E.) < or = 16%. Precise quantitation of PCR targets using co-amplification PCR requires multiple sample dilutions to ensure that the CMV native target sequence was in a close equimolar relationship with the CMV control sequence at the time of PCR amplification.

Base Sequence↗
PubMed1994

Highly cooperative and hysteretic response of the skeletal muscle ryanodine receptor to changes in proton concentrations.

Ryanodine receptors are key molecules in excitation-contraction coupling of skeletal muscle. They form the pore of the calcium release channel, which is regulated by Ca and ATP. Multiple proton titration sites are involved in controlling the different open states of the channel, as indicated by the following: i) the channel had a biphasic response to changes in proton concentrations around neutral pH; ii) the activities of the channel were inhibited by acidic pHs in a highly cooperative manner; and iii) the channel exhibited pronounced hysteresis to changes in pH. Four distinct conductance states can be identified in the single ryanodine-activated calcium release channel. The distribution of the multiple conductance states depends on the level of [Ca], ATP, and pH in the recording solution. The data are consistent with the multimeric structure of the skeletal muscle ryanodine receptor.

Adenosine Triphosphate↗
PubMed1994

Characterization of C-type natriuretic peptide receptors in human mesangial cells.

Our aim was to examine whether the human glomerulus was a target for C-type natriuretic peptide (CNP) and how A, B and C receptors of natriuretic peptides (ANPR-A, ANPR-B, ANPR-C) were distributed in glomerular mesangial and epithelial cells. CNP stimulated cyclic GMP production in cultured human mesangial and epithelial cells with similar threshold concentrations (1 nM) and maximum effects (basal value x 30 at 1 microM). In contrast, atrial natriuretic peptide (ANP) was only stimulatory in epithelial cells. [125I] CNP bound specifically to mesangial cells with a Kd of 0.47 nM and Bmax of 42 fmol/mg. Equilibrium of binding was obtained after four to five hours at +4 degrees C and nonspecific binding represented 10 to 20% of total binding. HS142-1 (100 micrograms/ml), a specific inhibitor of ANPR-A and ANPR-B, suppressed 90% of CNP-dependent cyclic GMP production whereas it had little effect on [125I]-CNP binding, suggesting that C receptors were largely predominant in mesangial cells. No biological effect of CNP on mesangial cells, including change in basal or angiotensin II-induced contractility and inhibition of basal or serum-dependent proliferation, could be demonstrated. Similar results were obtained with 8-bromo-cyclic GMP and sodium nitroprusside. Intraglomerular localization of ANPR-A, ANPR-B and ANPR-C mRNA was studied using reverse transcriptase-polymerase chain reaction with amplification of their corresponding cDNA by different primers. Amplification products were identified on gel electrophoresis by their predicted sizes and sequencing. ANPR-A, ANPR-B and ANPR-C mRNA were present in epithelial cells whereas only ANPR-B and ANPR-C mRNA were detected in mesangial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial Natriuretic Factor↗
PubMed1994

Relationship between different recovery curves representing rate-dependent AV nodal function in rabbit heart.

INTRODUCTION: The rate-dependent changes in atrioventricular (AV) nodal conduction time show different characteristics depending upon whether the conduction times are plotted against the atrial interval (AA-recovery curve) or His-atrial interval (HA-recovery curve). This study characterizes these differences in the context of controlled changes of nodal functional properties, determines their functional significance, and tests the hypothesis that they are related solely to the nodal conduction time of the last beat (last conduction time) before the premature beat. METHODS AND RESULTS: Premature nodal conduction times obtained in isolated rabbit heart preparations under various steady-state and transient conditions were plotted as a function of the corresponding HA and AA intervals, as well as the AA interval corrected for the last conduction time. Under all conditions, the corrected AA-recovery curve was indistinguishable in shape from the HA-recovery curve, and as such reflected similar underlying nodal functional properties. Moreover, a selective increase in the last conduction time, induced in the absence of time-dependent effects associated with the functional property of fatigue, shifted the AA-recovery but not the HA-recovery curve upward with respect to the control curve. CONCLUSION: The last conduction time accounts entirely for differences between AA-recovery and HA-recovery curves that otherwise reflect the same underlying nodal functional state. Thus, a consistent assessment of rate-dependent changes in nodal function can be achieved with either measure of recovery time.

Animals↗
PubMed1994

Mouse p53 represses the rat brain creatine kinase gene but activates the rat muscle creatine kinase gene.

The creatine kinases (CK) regenerate ATP for cellular reactions with a high energy expenditure. While muscle CK (CKM) is expressed almost exclusively in adult skeletal and cardiac muscle, brain CK (CKB) expression is more widespread and is highest in brain glial cells. CKB expression is also high in human lung tumor cells, many of which contain mutations in p53 alleles. We have recently detected high levels of CKB mRNA in HeLa cells and, in this study, have tested whether this may be due to the extremely low amounts of p53 protein present in HeLa cells. Transient transfection experiments showed that wild-type mouse p53 severely repressed the rat CKB promoter in HeLa but not CV-1 monkey kidney cells, suggesting that, in HeLa but not CV-1 cells, p53 either associates with a required corepressor or undergoes a posttranslational modification necessary for CKB repression. Conversely, mouse wild-type p53 strongly activated the rat CKM promoter in CV-1 cells but not in HeLa cells, suggesting that, in CV-1 cells, p53 may associate with a required coactivator or is modified in a manner necessary for CKM activation. The DNA sequences required for p53-mediated modulations were found to be within bp -195 to +5 of the CKB promoter and within bp -168 to -97 of the CKM promoter. Moreover, a 112-bp fragment from the proximal rat CKM promoter (bp -168 to -57), which contained five degenerate p53-binding elements, was capable of conferring p53-mediated activation on a heterologous promoter in CV-1 cells. Also, this novel p53 sequence, when situated in the native 168-bp rat CKM promoter, conferred p53-mediated activation equal to or greater than that of the originally characterized far-upstream (bp -3160) mouse CKM p53 element. Therefore, CKB and CKM may be among the few cellular genes which could be targets of p53 in vivo. In addition, we analyzed a series of missense mutants with alterations in conserved region II of p53. Mutations affected p53 transrepression and transactivation activities differently, indicating that these activities in p53 are separable. The ability of p53 mutants to transactivate correlated well with their ability to inhibit transformation of rat embryonic fibroblasts by adenovirus E1a and activated Ras.

Animals↗
PubMed1994

Coexisting beta-adrenoceptor subtypes: significance for thermogenic process in brown fat cells.

The possible significance of the coexisting beta 1-, beta 2-, and beta 3-adrenoceptors in brown adipose tissue for the thermogenic response was investigated. Oxygen consumption of isolated hamster brown fat cells was analyzed as a measure of thermogenesis. Thermogenesis could be evoked not only by the physiological agent norepinephrine but also by BRL-37344 and CGP-12177. No evidence for biphasic inhibition curves was found with either the selective beta 1-antagonist ICI-89406, the beta 2-antagonist ICI-118551, or the beta 1/beta 2-nonselective beta-antagonist propranolol against 1 microM norepinephrine; pI50 (the negative logarithm of the inhibitory constant for an antagonist, as estimated from the dose-response curve for an antagonist vs. a constant agonist concentration) values for ICI-89406 and ICI-118551 were very low (4-5), implying nonselective inhibition; the pI50 for propranolol was approximately 6 (as expected for the beta 3-receptor). Even with suboptimal norepinephrine, no biphasic inhibition was found. CGP-12177 at concentrations where it is primarily an antagonist to the beta 1-receptor did not influence the dose-response curve for either norepinephrine or BRL-37344. BRL-37344- or CGP-12177-induced thermogenesis was inhibited by the beta-antagonists in a manner similar to norepinephrine-induced thermogenesis. Schild plots for propranolol inhibition of norepinephrine-, isoprenaline-, BRL-37344- and CGP-12177-induced thermogenesis yielded similar pA2 (the negative logarithm of the inhibitory constant for an antagonist, as calculated from a series of agonist dose-response curves at different antagonist concentrations) (approximately 5.5), for interaction with either agonist, implying that the same receptor was stimulated by all agonists. Thus, despite the fact that different beta-receptor subtypes coexist in the tissue, we find no evidence for the participation of beta 1- or beta 2-receptors in the thermogenic response. Within the resolution of the experiments, the results therefore imply that it is predominantly or solely the beta 3-receptor that is coupled to thermogenesis, and it is via this beta-adrenergic receptor that not only norepinephrine but also CGP-12177 and BRL-37344 induce thermogenesis.

Adipose Tissue, Brown↗
PubMed1994

Production and characterization of transformed B-lymphocytes expressing the membrane defect of Scott syndrome.

Scott syndrome is a bleeding disorder associated with an isolated defect in expression of membrane coagulant activity by stimulated platelets. This defect represents a decrease in platelet membrane binding sites for coagulation factors Va and VIIIa, reflecting diminished surface exposure of phosphatidylserine (PS). To gain insight into the cellular and genetic basis for this disorder, B-lymphocytes from a patient with Scott syndrome and from normal donors were immortalized by EBV-transformation, and tested for their capacity to expose plasma membrane PS in response to the Ca2+ ionophore, A23187. Upon incubation with A23187, EBV-lymphoblasts derived from normal donors consistently induced surface expression of PS in > 70% of all cells, as detected by membrane association of the PS-binding proteins, factor Va or annexin V. PS exposure in these cells was maximal after 5 min, and saturated at < 100 microM external free [Ca2+]. By contrast, < 30% of Scott syndrome lymphoblasts exposed PS, and saturation was not observed at > 1 mM external free [Ca2+]. Single-cell clones derived from the Scott lymphoblasts all exhibited a diminished response to A23187 comparable with that of the parental cells, suggesting that all lymphocytes from this patient share this membrane abnormality. Hybridomas prepared by fusion of Scott lymphoblasts with the myeloma cell line UC-LUC showed responses to Ca2+ ionophore comparable to those observed for normal lymphoblasts and for hybridomas prepared by fusion of normal lymphoblasts with UC-LUC. This correction of the Scott abnormality suggests possible complementation of an aberrant gene(s) responsible for this disorder.

Annexin A5↗
PubMed1994

Disruption by interferon-alpha of an autocrine interleukin-6 growth loop in IL-6-dependent U266 myeloma cells by homologous and heterologous down-regulation of the IL-6 receptor alpha- and beta-chains.

IL-6 is an autocrine growth factor for U266 myeloma cells and their growth is inhibited by IFN-alpha or IL-6 mAb. We asked, therefore, whether IFN-alpha-induced growth inhibition involved IL-6. IFN-alpha and mAb against IL-6, the IL-6R alpha-(gp80) or beta-chain (gp130) potently inhibited U266 cells. Remarkably, this effect occurred despite IFN-alpha-augmented secretion of endogenous IL-6. However, examining the IL-6R revealed that IFN-alpha drastically curtailed expression of the IL-6R alpha- and beta-chain. This effect occurred on two different levels (protein and mRNA) and by two different mechanisms (directly and indirectly through IL-6). First, IFN-alpha, but not IL-6, greatly decreased gp80 and, to a lesser extent, gp130 mRNA levels which resulted in a loss of IL-6 binding sites. Second, IFN-alpha-induced IL-6 predominantly down-regulated membrane-bound gp130. IFN-alpha-mediated decrease of gp80 levels was not detected on IL-6-independent myeloma (RPMI 8226) or myeloid cells (U937). We conclude that IFN-alpha inhibited IL-6-dependent myeloma cell growth by depriving U266 cells of an essential component of their autocrine growth loop, a functional IL-6R.

Affinity Labels↗
PubMed1994

Modulation of progesterone-induced Xenopus oocyte maturation by prostaglandins E1 and E2.

Prostaglandins E1 (PGE1) and E2 (PGE2), the general local modulators or "local" hormones, were able to inhibit the progesterone-induced maturation of Xenopus oocytes in vitro, but could not induce maturation by themselves. This inhibition was observed on the sensitivity of the maturation process of the oocyte responses to progesterone, but not on the maximum response or responsiveness. The decreased sensitivity of oocytes to progesterone by prostaglandin E1 or E2 was evident in both the increased concentration of progesterone and the prolonged time required for the maturation of 50% of the oocyte. PGE1 and PGE2 did not appear to affect either the basal level or the progesterone-reduced cAMP level of oocytes. Microinjection of PGE1 or PGE2 into the oocytes had no effect on progesterone-induced oocyte maturation, suggesting that other cell surface-mediated signaling events might be responsible for this modulatory effect of the progstaglandins.

Alprostadil↗
PubMed1994

Effects of parathyroid hormone and estradiol on proliferation and function of human osteoblasts from fetal long bone. An in vitro study.

The effect of parathyroid hormone (PTH) (0.01 nM-10 nM) and 17 beta-estradiol (E2, 1 nmol-10 nM) alone or in combination on 3H-thymidine incorporation, alkaline phosphatase and adenylate cyclase activities were investigated in human fetal osteoblasts using serum-free monolayer primary cultures. The results showed that PTH inhibited cell proliferation while E2 promoted it. On alkaline phosphatase activity, PTH showed a complex results while E2 were slightly inhibitory. PHT-E2 combination suggested that E2 could alter the effect of PTH alone, also potentiated the anabolic and antagonize the catabolic effects of PTH on bone formation.

Adenylyl Cyclases↗
PubMed1994

[Epidemiological study on the infections of Mycoplasma and Chlamydia in immoral persons and healthy controls in seven areas of China].

1,284 immoral persons and 1,161 healthy controls, from seven areas of China, were detected for the antibodies to Ureaplasma urealyticum (Uu), Mycoplasma hominis (Mh), and Chlamydia trachomatis (Ct) by indirect hemoagglutination (IHA). The results showed that: (1) in the immoral persons, the positive rates of antibody was the highest in Uu (25.47%), higher in Ct (18.22%), and lower in Mh (8.80%); (2) the levels of antibody to Mh, Uu, and Ct were significantly higher in the immoral persons than in the healthy controls; and (3) the immoral persons were commonly complicated with infections caused by Uu and Ct. It was indicated that the immoral persons were risk population of Mycoplasma and/or Chlamydial veneral diseases, and it must be noted for us controlling veneral diseases.

Adolescent↗
PubMed1994

Farnesylation of p21 Ras proteins in Xenopus oocytes.

Unprocessed p21 Ras proteins microinjected into Xenopus oocytes were radiolabeled by coinjected [3H]farnesyl pyrophosphate, a direct farnesyl donor substrate for all known mammalian farnesyltransferases. Mevinolin, an inhibitor of HMG CoA reductase which reduces the levels of mevalonate and thus farnesyl pyrophosphate, blocked oncogenic H-Rasva112 induced germinal vesicle breakdown in oocytes. This mevinolin caused block was completely reversed by co-injected farnesyl pyrophosphate. The putative farnesyltransferase in Xenopus oocytes was identified to be similar to those found in mammalian cells in that it requires an intact CAAX box motif in addition to the conserved cysteine residue at the fourth position from the C-terminus of Ras proteins for its farnesylating activity. Peptide inhibitors of farnesyltransferase such as CVIM and TKCVIM were shown to inhibit farnesylation of microinjected Ras proteins thereby blocking its function namely the induction of oocyte maturation. These results demonstrate that Xenopus oocytes process bacterially produced mammalian Ras proteins in a manner similar to, if not identical with that in mammalian cells, thus validating the continued use of the Xenopus oocyte system for unraveling the functions of Ras proteins. Furthermore, our results indicate that the oocyte system may be a useful in vivo model for studying the farnesylation of human Ras proteins, its regulation, and the effects of farnesyltransferase inhibitors.

Alkyl and Aryl Transferases↗
PubMed1994

[Investigation of sites of magnetic coil placement and its influence on CMAPs].

Varying CMAP results with changing coil location during magnetic stimulation of facial nerve have been noted in the literature. In order to investigate parameters for clinical application, 20 normal subjects were tested with Magstim Model 200 Magnetic Stimulator synchronized with Nicolet Viking II with 9 designed points of coil location at the parietal-occipital-temporal area. The amplitude and latency were recorded and analyzed statistically. Results demonstrated that the maximal amplitude was mainly obtained from following 3 points: C (midpoint between vertex and external auditory canal), V-2 (2 cm inferior to C) and H-2 (2 cm posterior to C), which showed the best location of coil for magnetic stimulation among the tested points. The maximal amplitude was also observed in points of H-4 (4 cm posterior to C) and V+2 (2 cm superior to C) in 4 subjects. In contrast to the varying amplitude, latency obtained from each stimulating point was constant, so we postulate that the same segment of facial nerve was excited regardless of moving magnetic coil around those points.

Action Potentials↗