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Biomedical subjects

Z Cai

Publications and source records attributed to Z Cai.

At least 145 records · Page 8Linked to original sources
PubMed1997

IkappaB alpha overexpression in human breast carcinoma MCF7 cells inhibits nuclear factor-kappaB activation but not tumor necrosis factor-alpha-induced apoptosis.

Nuclear factor-kappaB (NF-kappaB) is one of major component induced by tumor necrosis factor-alpha (TNF), and its role in the signaling of TNF-induced cell death remains controversial. In order to delineate whether the involvement of NF-kappaB activation is required for triggering of the apoptotic signal of TNF, we inhibited the nuclear translocation of this transcription factor in TNF-sensitive MCF7 cells by introducing a human MAD-3 mutant cDNA coding for a mutated IkappaB alpha that is resistant to both phosphorylation and proteolytic degradation and that behaves as a potent dominant negative IkappaB alpha protein. Our results demonstrated that the mutated IkappaB alpha was stably expressed in the transfected MCF7 cells and blocked the TNF-induced NF-kappaB nuclear translocation. Indeed, TNF treatment of these cells induced the proteolysis of only the endogenous IkappaB alpha but not the mutated IkappaB alpha. The nuclear NF-kappaB released from the endogenous IkappaB alpha within 30 min of TNF treatment was rapidly inhibited by the mutated IkappaB alpha. There was no significant difference either in cell viability or in the kinetics of cell death between control cells and the mutated IkappaB alpha transfected cells. Furthermore, electron microscopic analysis showed that the cell death induced by TNF in both control and mutated IkappaB alpha transfected cells was apoptotic. The inhibition of NF-kappaB translocation in mutated IkappaB alpha-transfected cells persisted throughout the same time course that apoptosis was occurring. Our data provide direct evidence that the inhibition of NF-kappaB did not alter TNF-induced apoptosis in MCF7 cells and support the view that TNF-mediated apoptosis is NF-kappaB independent.

Apoptosis↗
PubMed1997

Glutamate receptor-mediated calcium responses in acutely isolated hippocampal astrocytes.

GFAP(+) cells were acutely isolated from the hippocampi of 1-10 day old rats, and the intracellular calcium responses to L-glutamate, ATP, and 5-HT were studied in single cells. Eighty-two percent of such cells responded to glutamate, 20% of them responded to ATP, and none responded to 5-HT. The same cells that failed to respond to ATP and 5-HT often responded to glutamate. These proportions of cells responding to glutamate and ATP are very similar to those reported for GFAP(+) astrocytes in hippocampal slices (Porter and McCarthy, 1995a,b). After culturing for 1-2 days in serum-containing medium, 60% of such acutely isolated cells responded to either glutamate or ATP, and 5% to 5-HT. After 1 week in culture, the percentage of cells responding to glutamate remained essentially the same (62%) but the percentages of cells responding to ATP and 5-HT increased to 92% and 62%, respectively. These percentages were very close to the results obtained from primary hippocampal astrocyte cultures prepared from 1 day old rats and cultured for 1-2 weeks in vitro. Pharmacological characterization showed that the Ca2+ responses of acutely isolated hippocampal astrocytes from P1-10 rats was due to activation of a group I metabotropic glutamate receptor. The calcium responses to ATP and 5-HT in acutely isolated cells that were then cultured were mediated by P2y and 5-HT2A receptors, respectively. These data show that, like cortical astrocytes (Kimelberg et al., 1997), GFAP(+) astrocytes cultured from the hippocampi of young rats showed marked differences in receptor expression compared to their acutely isolated counterparts. Also, since the astrocytes acutely isolated from these 2 different brain regions showed qualitatively the same responses for the 3 receptors selected, it indicates a degree of homogeneity of receptor expression for astrocytes from these 2 brain regions.

Adenosine Triphosphate↗
PubMed1997

Transmission electron microscopic investigation of high-palladium dental casting alloys.

OBJECTIVE: The purpose of this study was to use transmission electron microscopy to examine four representative high-palladium alloys and gain insight into possible strengthening mechanisms. METHODS: Castings of two Pd-Cu-Ga alloys and two Pd-Ga alloys were thinned by jet polishing and ion milling, followed by plasma cleaning, to yield foil specimens. Multiple specimens were prepared for each alloy. Bright-field images, dark-field images and selected-area electron diffraction patterns for the alloys in the as-cast condition, after simulated porcelain-firing heat treatment, and after annealing at 980 degrees C were analyzed by standard transmission electron microscope (TEM) techniques. The overall compositions of the ultrastructures for the specimen foils were determined by conventional standardless energy-dispersive spectroscopic analyses with the TEM, and mean values of the elemental compositions were compared to the nominal alloy compositions provided by the manufacturers. RESULTS: There was generally good agreement (differences less than 2 wt%) between the overall ultrastructure composition and each nominal alloy composition, except for Protocol from which in may have been lost during casting or formed intermetallic compounds that were not detected by TEM. The same fine-scale tweed structure within parallel bands of approximately 100-200 nm width was observed for all four alloys in the as-cast condition and after simulated porcelain-firing heat treatment. The persistence of the ultrastructure in the specimens of the two Pd-Cu-Ga alloys annealed at 980 degrees C and quenched in ice water indicated very rapid formation from the palladium solid solution. The presence of ¿100¿ and ¿110¿ forbidden reflections for the <001> zone suggested that the tweed structure is ordered, although further research is necessary to establish this conclusion. SIGNIFICANCE: The presence of a similar tweed structure in both the Pd-Cu-Ga alloys and the Pd-Ga alloys of substantially lower hardness shows that some other strengthening mechanism accounts for the high hardness and strength generally observed for Pd-Cu-Ga alloys.

Copper↗
PubMed1997

Implication of an increased oxidative stress in the formation of advanced glycation end products in patients with end-stage renal failure.

Recent studies have demonstrated a marked increase in the level of advanced glycation end products (AGEs) in the plasma, skin and amyloid fibrils of hemodialysis (HD) patients. The presence of AGEs in (beta2m) forming amyloid fibrils has been established in a previous immunochemical study relying on a monoclonal anti-AGE antibody. In the present study, Western blot analysis and immunohistochemistry reveal that the epitope recognized by this antibody is N epsilon-(carboxymethyl)lysine (CML) and that CML is one of the AGE structures present in amyloid fibrils. Thus, two AGE structures, CML and pentosidine, are now recognized in dialysis-related amyloidosis. AGE accumulation in uremia is not accounted for by elevated glucose levels. Since CML and pentosidine formation are closely linked to oxidative processes, we tested the hypothesis that a high oxidative stress enhanced AGE formation in HD patients. We focused on ascorbic acid (AA) because AA is easily oxidized under oxidative stress and its oxidized form (oxiAA) is a source of CML and pentosidine. In vitro incubation of beta2m with AA under atmospheric oxygen resulted in: (1) the rapid appearance of characteristic physicochemical properties of AGEs (brown color, fluorescence, polymerization tendency); (2) the transformation of beta2m into AGE-modified beta2m recognized by a specific monoclonal antibody; and (3) the accelerated formation of CML in beta2m and beta2m-peptide, recognized by mass spectrometry. A similar in vitro incubation of human serum albumin disclosed a parallel production of pentosidine measured by high-performance liquid chromatographic assay. In HD patients, the degree of AA oxidation, assessed as the ratio of oxiAA to total ascorbate, was more than twice as high as that of normal subjects (0.87 +/- 0.16 vs. 0.35 +/- 0.11, P < 0.0001), suggesting the presence of an increased oxidative stress. Interestingly, plasma level of oxiAA was correlated with the plasma levels of protein linked (P < 0.01, r2 = 0.25) and free (P < 0.05, r2 = 0.22) pentosidine. Altogether these results demonstrate that AGE, that is, CML and pentosidine, production is accelerated under oxidative stress, even in the absence of glucose. They suggest that, in uremia, CML and pentosidine production is determined both by an increased oxidative stress and the availability of precursors such as oxiAA. Finally, both CML and pentosidine contribute to the AGEs present in dialysis-related amyloid fibrils.

Amino Acid Sequence↗
PubMed1997

Transmitter-induced calcium responses differ in astrocytes acutely isolated from rat brain and in culture.

Glial fibrillary acid protein (GFAP)-positive astrocytes isolated from the cerebral cortices of 3-10-day-old rats frequently showed increased intracellular Ca2+ concentration responses to L-glutamate and glutamate analogues. However, few of the acutely isolated cells responded to ATP, and no such cells responded to serotonin [5-hydroxytryptamine (5-HT)]. The same cell that failed to respond to ATP or 5-HT often responded to glutamate. Culturing acutely isolated cells in media containing horse serum decreased Ca2+ responses to glutamate but increased the responses to ATP and induced responses to 5-HT. In primary cultures prepared from the cerebral cortices of 1-day-old rats and cultured in horse serum, fewer of the cells responded to glutamate, but almost all cells responded to ATP and 5-HT. The lack of or limited response to, 5-HT or ATP in the acutely isolated cells seems unlikely to be due to selective damage to the respective receptors because acutely isolated GFAP-negative cells showed responses to ATP, several different proteases and mechanical dissociation yielded cells that also responded to glutamate but not to ATP, and exposure of primary cultures to papain did not abolish Ca2+ responses to several transmitters. The responses of the acutely isolated cells to glutamate but limited or lack of responses to ATP and 5-HT also correspond to what has been seen so far for astrocytes in situ. Thus, the present studies provide direct evidence that some of the receptors seen in primary astrocyte cultures may reflect a response to culture conditions and that, in the context of the relevant information so far available, acutely isolated astrocytes seem to reflect better the in vivo state.

Adenosine Triphosphate↗
PubMed1997

[Treatment of atrial fibrillation using maze procedure by radiofrequency ablation].

From May 1994 to May 1996, 20 cases of atrial fibrillation were treated by means of Maze procedure by radiofrequenncy ablation, at the same time 19 cases of these patients were complicated with rheumatic heart valve disease and valve replacement operations were perfomned, in the other case atrial septal defect was repaired. Yoshio Kosakai's operation route was adopted in radiofrequency ablation procedure. After operation 16 patients of atrial fibrillation resumed sinus rhythm (80%), in 4 casess of atrial fibrillation sinus rhythm was unsuccessfully restored, two patients remained atrial fibrillation, one patient was of atrial flutter, the other was of nodal rhythm. Short time was needed in radiofrequency ablation Maze procedure, average time increase of aortic clamping was 20.5 minutes, and there was no danger of hemorrhage related to this kinds of Maze procedure. During 7-10 days after operation, there appeared superventricular arrhythmia which might be related to ill-distribution of radiofrequency ablation, and interference of atrial electric activity.

Adult↗
PubMed1997

[Histopathological study on traumatic facial nerve injury in the rabbits].

Six patterns of injuries (exposure, compression, crush, stretch, division and post-division anastomosis) were analysed in 136 facial nerve buccal branches of 68 rabbits in order to establish the experimental model for study of traumatic facial nerve injury. Changes of histopathology at definite time were studied. The results showed that: exposure can create degree I injury; compression, stretch and crush can create degree II-III injury; the prognosis in anastomosis group was obviously better than unanastomosis group. In moderate to serious injuries, changes of histopathology and ENoG were synchronized, but were not so in mild and later stage of serious injury. The 15th day postinjury was the best time of estimating the facial nerve function clinically. The area of injury would obviously influence the mylin lesion, while the quality and degree of injury would obviously influence the axonal lesion.

Animals↗
PubMed1997

Stable expression of human cytochrome CYP2B6 and CYP1A1 in Chinese hamster CHL cells: their use in micronucleus assays.

With specific designed primers, CYP2B6 and CYP1A1 cDNA were generated by reverse transcription-polymerase chain reaction(RT-PCR) technique performed on total RNAs isolated from human liver and 3-methylcholanthrene(3-MC) induced human amnion FL cells. Cell lines (CHL-2B6 and CHL-1A1) capable of expressing human cytochrome P450(CYP)2B6 and 1A1 were established after transfection of corresponding eukaryotic recombinant expression plasmid with human CYP2B6 and 1A1 cDNA inserts respectively. These cell lines stably expressed the mRNAs and the enzymatic activities corresponding to CYP2B6 and CYP1A1, respectively. Compared with Chinese hamster 1ung(CHL) cells, the micronucleus frequency in CHL-2B6 cells is markedly increased when exposed to nitrosamines, aflatoxin B1(AFB1) and cyclophosphamide (CPA). This is also in CHL-1A1 cells, when exposed to carcinogenic polycyclic aromatic hydrocarbons.

Animals↗
PubMed1997

Improved methods for making and using glass knives.

We have observed over time that the right side of a glass knife is the optimal cutting edge for microtomy if the counterpiece (heel opposite the edge) is controlled within 1 mm. The right cutting edge has been considered the "saw toothed" side and has not been used for ultrathin sectioning. We have observed that the right cutting edge is sharper and more durable than the left. Light and scanning electron microscopy were used to observe the cutting edge, and transmission electron microscopy was used to examine semithin and ultrathin sections of animal and plant tissues cut by the right and left sides of the cutting edge. The results indicate that the cutting edge becomes sharper and more durable from left to right. Both the quality and efficiency of ultrathin sectioning is improved by using the right cutting edge.

Animals↗
PubMed1996

Transfected Drosophila cells as a probe for defining the minimal requirements for stimulating unprimed CD8+ T cells.

Stimulation of naive T cells by antigen-presenting cells (APC) is thought to involve two qualitatively different signals: signal one results from T-cell receptor (TCR) recognition of antigenic peptides bound to major histocompatibility complex (MHC) molecules, whereas signal two reflects contact with one or more costimulatory molecules. The requirements for stimulating naive T cells were studied with MHC class I-restricted CD8+ T cells from a T-cell receptor transgenic line, with defined peptides as antigen and transfected Drosophila cells as APC. Three main findings are reported. First, stimulation of naive T cells via signal one alone (MHC plus peptide) was essentially nonimmunogenic; thus T cells cultured with peptides presented by MHC class I-transfected Drosophila APC lacking costimulatory molecules showed little or no change in their surface phenotype. Second, cotransfection of two costimulatory molecules, B7-1 and intercellular adhesion molecule 1 (ICAM-1), converted class I+ Drosophila cells to potent APC capable of inducing strong T-proliferative responses and cytokine (interleukin 2) production. Third, B7-1 and ICAM-1 acted synergistically, indicating that signal two is complex; synergy between B7-1 and ICAM-1 varied from moderate to extreme and was influenced by both the dose and affinity of the peptide used and the parameter of T-cell activation studied. Transfected Drosophila cells are thus a useful tool for examining the minimal APC requirements for naive T cells.

Animals↗
PubMed1996

Impairment of Fas-antigen expression in adriamycin-resistant but not TNF-resistant MCF7 tumor cells.

Anti-cancer drugs and cytotoxic cytokines such as members of the TNF/Fas-ligand family play a predominant role in apoptosis induction in tumor cells, and are critical in cancer therapy. In this study we used the human breast-carcinoma cell line MCF7, its derivatives MCF7Adr (resistant to adriamycin) and R-A1 (resistant to TNF), to determine the impact of acquired drug and cytokine resistance on susceptibility to Fas-induced cytotoxicity and Fas-antigen expression. While MCF7 and R-A1 cells were killed by anti-Fas in the presence of IFN-gamma, MCF7Adr was found to be resistant to Fas-mediated apoptosis. This resistance was correlated with a loss of surface Fas-protein expression. Fas-gene transfer in MCF7Adr resulted in high sensitivity to Fas-mediated cytotoxicity, indicating that the Fas signalling pathway is virtually intact in this cell line. Over-expression of the MDR1 gene in MCF7 following gene transfer did not affect Fas expression and anti-Fas sensitivity, suggesting that the P-gp-mediated multidrug-resistance phenotype is not directly involved in the loss of Fas expression, contrary to what has been observed by others in T-cell lines. Furthermore, the down-regulation of Fas expression and subsequent resistance to anti-Fas were observed in drug-resistant human ovarian-carcinoma IGR-OV1/VCR cells and leukemic lymphoblast CEM/VLB cells, suggesting that the alteration of Fas expression following drug-resistance selection is not restricted to one cell type.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

Differing roles for B7 and intercellular adhesion molecule-1 in negative selection of thymocytes.

To ensure self tolerance, immature thymocytes with high binding affinity for self peptides linked to major histocompatibility complex (MHC) molecules are eliminated in situ via apoptosis (negative selection). The roles of two costimulatory molecules, B7-1 and intercellular adhesion molecule-1 (ICAM-1), in negative selection was examined by studying apoptosis of T cell receptor transgenic CD4+8+ thymocytes cultured with specific peptides presented by MHC class I-transfected Drosophila cells. When coexpressed on these cells, B7-1 and ICAM-1 act synergistically and cause strong class 1-restricted negative selection of thymocytes. When expressed separately, however, B7-1 and ICAM-1 display opposite functions: negative selection is augmented by B7-1, but is inhibited by ICAM-1. It is notable that B7-1 is expressed selectively in the thymic medulla, whereas ICAM-1 is expressed throughout the thymus. Because of this distribution, the differing functions of B7-1 and ICAM-1 may dictate the sites of positive and negative selection. Thus, in the cortex, the presence of ICAM-1, but not B7-1, on the cortical epithelium may preclude or reduce negative selection and thereby promote positive selection. Conversely, the combined expression of B7-1 and ICAM-1 may define the medulla as the principal site of negative selection.

Amino Acid Sequence↗
PubMed1996

Intrauterine hypoxia-ischemia reduces phosphoinositide hydrolysis stimulated by metabotropic glutamate receptor agonists in cultured rat cerebellar granule cells.

Effects of intrauterine hypoxia-ischemia (HI) on receptor-stimulated phosphoinositide (PPI) hydrolysis were studied in rat cerebellar granule cell cultures prepared from an in utero HI model. On gestation day 17, HI conditions were achieved by complete clamping of the uterine vasculature for 30 min followed by removal of the clamps to permit reperfusion. Sham operation (SH, surgery without vasculature ligation) was performed as the control. Intrauterine HI did not affect the basal level of PPI hydrolysis (in the absence of stimulants) in cells prepared from either the SH or the HI group. PPI hydrolysis stimulated by quisqualate (QA) or trans-(1S,3R)-1-amino-1,3-cyclo-pentanedicarboxylic acid (trans-ACPD) was significantly reduced in cells prepared from the HI group, whereas intrauterine HI did not affect the PPI hydrolysis induced by ionotropic glutamate receptor agonists or by norepinephrine or serotonin. At a dose range of 100-300 microM, QA-stimulated PPI hydrolysis in cells prepared from the SH group increased by 3-to 4.5-fold, while this increase was only 2- to 2.5-fold in cells prepared from the HI group. Presence of L-NG-monomethyl-arginine (L-NMMA), a nitric oxide (NO) synthase inhibitor, did not increase QA-stimulated PPI hydrolysis in cells prepared from either the SH or the HI group, indicating that stimulation of NO formation is unlikely involved in the suppressive effects of intrauterine HI on QA-induced PPI hydrolysis. The QA-stimulated PPI hydrolysis in cells prepared from the HI group, but not from the SH group, was further inhibited by L-(+)-2-amino-3-phosphono-propionic acid (L-AP3). The overall results suggest that intrauterine HI has long-lasting suppressive effects on metabotropic glutamate receptor agonist-stimulated PPI hydrolysis and these effects might be associated with alterations in expression of metabotropic glutamate receptor subtypes.

Alanine↗
PubMed1996

Solution structure of loop A from the hairpin ribozyme from tobacco ringspot virus satellite.

The solution structure of loop A from the hairpin ribozyme found in the minus strand of tobacco ringspot virus satellite has been determined by NMR spectroscopy. The ribozyme consists of two internal loops flanked by short helices: loop A and helices I and II include the substrate and substrate binding site; loop B and helices III and IV are the catalytic domain. Loop A is a symmetric internal loop of eight nucleotides that contains the cleavage site. The 2-amino group of the guanine immediately 3' to the cleavage site is essential for catalysis. NMR results show that this guanine forms a sheared G.A base pair. The cytosine residue immediately 5' to the cleavage site forms an AH+.C base pair with an adenine whose pKa is shifted to 6.2 to allow partial protonation near neutral pH. Although the residues flanking the cleavage site are stacked in an A-form pattern, the phosphodiester backbone next to the cleavage site on the 3' side is splayed apart. This places the following base-a uracil-in the expanded major groove. The conformational flexibility and the lack of steric hindrance of the uracil as well as the unoccupied Watson-Crick positions on the sheared G.A base pair can allow loop A to specifically interact with the catalytic domain (loop B) without drastically changing its own conformation. The three-dimensional structure of loop A provides explanations for previously published mutation and structural mapping results.

Base Sequence↗
PubMed1996

Influence of antigen dose and costimulation on the primary response of CD8+ T cells in vitro.

The influence of costimulation on the primary response of CD8+ T cells to class I alloantigens was studied with the aid of a T cell receptor transgenic model and defined peptides as antigen. With small doses of antigen, the proliferative response of CD8+ cells was high early in culture but was of brief duration and declined to low levels by day 4; this abbreviated response was associated with limited production of interleukin 2 (IL-2) and was strongly dependent upon costimulation via CD8-major histocompatibility complex class I and CD28-B7 interactions. The response to large doses of antigen was quite different in two respects. First, large doses of antigen inhibited the early (day 3) proliferative response but caused a marked elevation of the response late in culture (day 5); these altered kinetics were associated with increased production of IL-2. Second, the initial proliferative response to large doses of antigen did not require costimulation: indeed, blocking costimulation with CTLA4lg or anti-CD8 monoclonal antibody enhanced the early proliferative response. However, blocking costimulation impaired IL-2 production and prevented the late proliferative response. These findings indicate that the requirement for costimulation of T cells can be partly overcome by increasing the dose of antigen to a high level. However, costimulation plays a key role in prolonging the response, presumably by triggering strong and sustained production of IL-2.

Abatacept↗
PubMed1996

Fas-mediated apoptosis in cultured human eosinophils.

Previous studies have shown that cytokine-dependent eosinophils undergo apoptosis, yet the mechanisms governing this phenomenon remain obscure. Fas antigen is a transmembrane glycoprotein belonging to the tumor necrosis factor receptor family. Cross-linking of Fas antigen in numerous cell types leads to apoptosis. In the present study, we examined the potential role of Fas antigen in the apoptosis of purified blood eosinophils from healthy donors. Cytokine-deprived eosinophils exhibited a time-dependent loss in viability, accompanied by an increase in the number of apoptotic nuclei and in the expression of Fas antigen and its mRNA, as shown by flow cytometry and reverse transcriptase-polymerase chain reaction, respectively. Cross-linking of Fas antigen with an agonistic anti-Fas monoclonal antibody (MoAb) induced a dose- and time-dependent increase in the number of apoptotic nuclei. Furthermore, using an in vitro coculture system, we showed engulfment of anti-Fas MoAb-treated eosinophils by monocyte-derived macrophages. Finally, incubation of eosinophils with the corticosteroid, dexamethasone, induced apoptosis and augmented that triggered by anti-Fas MoAb. Together, these observations suggest that Fas antigen expression and activation is involved in the apoptosis of human eosinophils and may contribute to the resolution of inflammatory allergic reactions in which eosinophil accumulation is a prominent feature.

Apoptosis↗