Biomedical subjects

L Zheng

Publications and source records attributed to L Zheng.

At least 19 recordsLinked to original sources

PubMed1995

Induction of apoptosis in mature T cells by tumour necrosis factor.

T-cell receptor-induced apoptosis regulates immune responses and can result from interactions between Fas (Apo1/CD95) and Fas ligand (FasL). Mutations in the genes for Fas and FasL cause disorders resembling human autoimmune diseases in lpr and gld mice, respectively. However, peripheral T-cell deletion takes place in lpr mice, and autoimmune syndromes occur in mouse strains without Fas or FasL defects. Here we show that tumour necrosis factor (TNF) can mediate mature T-cell receptor-induced apoptosis through the p75 TNF receptor. Blockage of both TNF and FasL is required to abrogate T-cell death and TNF mediates the death of most CD8+ T cells, whereas FasL mediates the death of most CD4+ T cells. Our results suggest that autoregulatory apoptosis of the mature T cells can occur by two distinct molecular mechanisms.

Animals↗

PubMed1995

Analysis of the CD4 coreceptor and activation-induced costimulatory molecules in antigen-mediated mature T lymphocyte death.

We have compared the signaling requirements for activation and lymphokine production in mature T lymphocytes to those required for TCR-driven programmed cell death (PCD). Both processes require TCR engagement and ligation of the CD4 coreceptor in the case of a T cell clone that recognizes Ag in the context of an MHC class II molecule. By contrast, stimulation through the CD28/B7 pathway does not appear to positively or negatively influence TCR-induced PCD, although it was required for IL-2 production in both resting and proliferating T cells. T cells that had been activated and induced to proliferate with IL-2 were found to express high levels of IL-2 mRNA upon TCR rechallenge, without a requirement for accessory cells. This was due to a strong up-regulation of the B7-1 molecule, but not the B7-2 molecule, on the T cell surface. These T cells that strongly costimulate each other are highly susceptible to TCR-induced death providing independent evidence that costimulatory signals are not protective. Thus, these results provide evidence that in mature T cells there exists a difference in the requirement for CD28 to achieve activation and IL-2 production compared with TCR-mediated PCD.

Abatacept↗

PubMed1995

Role of protein kinase C isozymes in Fc gamma receptor-mediated intracellular killing of Staphylococcus aureus by human monocytes.

Intracellular killing of Staphylococcus aureus by human monocytes after cross-linking Fc gamma R is known to be a phospholipase C (PLC)-dependent process. Activation of PLC leads to the formation of second messengers that synergistically activate protein kinase C (PKC). The aim of this study was to obtain more insight into the role of PKC in Fc gamma R-mediated killing process. PKC inhibitors H-7 and staurosporine markedly suppressed the killing of S. aureus by monocytes stimulated by cross-linking Fc gamma RI or -II. Cross-linking Fc gamma R caused a transient increase in PKC activity in the membranes of monocytes, as measured by Ca2+/phospholipid-dependent phosphorylation of histone. Western blot analysis revealed that cross-linking Fc gamma R stimulated a transient increase in PKC-beta in the membranes of monocytes with kinetics that correlated closely with the translocation of PKC activity. Cross-linking Fc gamma R on monocytes also stimulated the translocation of PKC-epsilon but not PKC-alpha. PMA and 1-oleoyl-2-acetylglycerol (OAG), which caused translocation of PKC-alpha, -beta, and -epsilon, did not stimulate the killing process. Incubation with these PKC activators for 10 min rendered monocytes unresponsive to stimulation of killing of S. aureus via Fc gamma R. It could be that activation of certain PKC isozymes, probably PKC-alpha and -epsilon, by these activators causes feedback inhibition of PLC and, consequently, the killing in monocytes, because PMA blocks the Fc gamma R-mediated intracellular inositol(1,4,5)P3 formation and PKC translocation. Together, our results indicate that PKC isozymes play an important role in both stimulation and inhibition of the Fc gamma R-mediated intracellular killing of bacteria by monocytes.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1995

Two genes encoding midgut-specific maltase-like polypeptides from Anopheles gambiae.

Full-length cDNA clones of two genes have been isolated from the African malaria vector mosquito, Anopheles gambiae. These genes, designated Agm1 and Agm2, encode maltase-like polypeptides of 498 and 599 residues, respectively. Deduced amino acid sequences contain a putative signal peptide sequence and four potential glycosylation sites. Agm1 and Agm2 show highest similarities to the Mal1 gene from Aedes aegypti and three clustered maltase genes from Drosophila melanogaster. Both genes are located at position 46D, in the terminal division of the left arm of the third chromosome. Agm2 has very strict tissue and temporal specificity, being expressed exclusively in the adult midgut. The specificity of Agm1 is similar but appears slightly broader; transcripts of this gene are detected at a low level in the pupae, and occasionally in the adult carcass after removal of the midgut.

Amino Acid Sequence↗

PubMed1995

MCML--Monte Carlo modeling of light transport in multi-layered tissues.

A Monte Carlo model of steady-state light transport in multi-layered tissues (MCML) has been coded in ANSI Standard C; therefore, the program can be used on various computers. Dynamic data allocation is used for MCML, hence the number of tissue layers and grid elements of the grid system can be varied by users at run time. The coordinates of the simulated data for each grid element in the radial and angular directions are optimized. Some of the MCML computational results have been verified with those of other theories or other investigators. The program, including the source code, has been in the public domain since 1992.

Absorption↗

PubMed1995

Mechanisms of transport of quinapril in Caco-2 cell monolayers: comparison with cephalexin.

PURPOSE: To determine the transport mechanisms of quinapril and cephalexin in Caco-2 cell monolayers, a cell culture model of the human small intestinal epithelium. METHODS: Uptake, transepithelial transport and intracellular accumulations of these two drugs were measured using Caco-2 cell monolayers grown onto Millicells and magnetically stirred diffusion chambers. RESULTS: Transepithelial transport, apical (AP)4 uptake and intracellular accumulation of both drugs depended on the maintenance of a transepithelial proton gradient and temperature of the medium. However, quinapril transport and accumulation, which did not display a maximum at approximately pH 6, was more sensitive to proton gradient change, whereas cephalexin transport was more sensitive to concentration change (range 0.5-5 mM). In addition, quinapril (1 mM) transport was decreased significantly (p < 0.05) by 10 mM cephalexin, loracarbef, Gly-Pro and Phe-Pro, but not by enalapril; whereas cephalexin (0.1 mM) transport was decreased significantly (p < 0.05) by all four compounds. Similarly, AP quinapril (1 mM) uptake was also decreased by 10 mM loracarbef, Gly-Pro, cephalexin, and enalapril, but these inhibitory effects (20-50%) were quantitatively less than their inhibitory effects on cephalexin uptake (50-90%). Finally, the AP uptake of quinapril was also significantly (p < 0.05) inhibited by FCCP (10 micrograms/ml), amiloride (0.5 mM), DEP (0.5 mM), and staurosporine (5 nM). CONCLUSIONS: The transport of quinapril in the Caco-2 cells is via a combination of the carrier-mediated proton gradient-dependent peptide transporter and passive diffusion.

Amino Acid Sequence↗

PubMed1995

Cyclosporin A, rapamycin and FK506 decrease prolactin release from rat pituitary cells in primary culture.

It is at present well established that prolactin exerts a non-specific immunoactivating function. In this work we tested whether the immunosuppressant drugs cyclosporin A, FK506 and rapamycin influence prolactin release from rat pituitary cells in primary culture. The tested drugs had no effect on the prolactin release measured during a 2h incubation period, indicating that they do not influence the secretion of prolactin from intracellular stores into the culture medium. During longer incubation times (48h), however, prolactin release was diminished to 56% +/- 18 (10 microM cyclosporin A), 64% +/- 14 (1 microM rapamycin) or 64% +/- 7 (1 microM FK506), suggesting an effect on prolactin production. At these drug concentrations no toxic effects were observed. The data indicate that inhibition of pituitary prolactin synthesis might contribute to the immunosuppressant action of cyclosporin A, rapamycin and FK506.

Animals↗

PubMed1995

Dependence of stimulus-transcription coupling on phospholipase D in agonist-stimulated pituitary cells.

Stimulation of phospholipase D activity is frequently observed during agonist activation of Ca(2+)-mobilizing receptors, but the cellular functions of this signaling pathway are not well defined. Pituitary gonadotrophs express Ca(2+)-mobilizing receptors for gonadotropin-releasing hormone (GnRH) and endothelin (ET), activation of which stimulates luteinizing hormone secretion and transient expression of c-fos. In pituitary cells and alpha T3-1 gonadotrophs, GnRH action was associated with both initial and sustained diacylglycerol (DG) production, whereas ET-1 induced only a transient DG response. Also, phospholipase D activity, estimated by the production of phosphatidylethanol from phosphatidylcholine in the presence of ethanol, was stimulated by GnRH but not ET-1. Such formation of phosphatidylethanol at the expense of phosphatidic acid (PA) during GnRH-induced activation of phospholipase D significantly reduced the production of PA, DG, and cytidine diphosphate diacylglycerol. Inhibition of PA-phosphohydrolase activity by propranolol also decreased GnRH-induced DG production and, in contrast to ethanol, increased PA and cytidine diphosphate diacylglycerol levels. The fall in DG production caused by ethanol and propranolol was accompanied by inhibition of GnRH-induced c-fos expression, whereas agonist-induced luteinizing hormone release was not affected. In contrast to their inhibitory actions on GnRH-induced early gene expression, neither ethanol nor propranolol affected ET-1-induced c-fos expression, or GnRH- and ET-1-induced inositol trisphosphate/Ca2+ signaling. These findings demonstrate that phospholipase D participates in stimulus-transcription but not stimulus-secretion coupling, and indicate that DG is the primary signal for this action.

Animals↗

PubMed1995

Temporal and spatial expression of amygdalin hydrolase and (R)-(+)-mandelonitrile lyase in black cherry seeds.

In black cherry (Prunus serotina Ehrh.) macerates, the cyanogenic diglucoside (R)-amygdalin undergoes stepwise degradation to HCN catalyzed by amygdalin hydrolase (AH), prunasin hydrolase, and (R)-(+)-mandelonitrile lyase (MDL). A near full-length AH cDNA clone (pAH1), whose insert encodes the isozyme AH I, has been isolated and sequenced. AH I exhibits several features characteristic of beta-glucosidases of the BGA family, including their likely nucleophile center (isoleucine-threonine-glutamic acid-asparagine-glycine) and acid catalyst (asparagine-glutamic acid-proline/isoleucine) motifs. The temporal expression of AH and MDL in ripening fruit was analyzed by northern blotting. Neither mRNA was detectable until approximately 40 days after flowering (DAF), when embryos first became visible to the naked eye. Both mRNAs peaked at approximately 49 DAF before declining to negligible levels when the fruit matured (82 DAF). Taken together with enzyme activity data, these time courses suggest that AH and MDL expression may be under transcriptional control during fruit maturation. In situ hybridization analysis indicated that AH transcripts are restricted to the procambium, whereas MDL transcripts are localized within cotyledonary parenchyma cells. These tissue-specific distributions are consistent with the major locations of AH and MDL protein in mature seeds previously determined by immunocytochemistry (E. Swain, C.P. Li, and J.E. Poulton [1992] Plant Physiol 100:291-300).

Aldehyde-Lyases↗

PubMed1995

Microsatellite DNA and isozyme variability in a west African population of Anopheles gambiae.

Microsatellites are defined as tracts of tandemly repeated short DNA sequences. Polymorphisms in this class of DNA are currently being used to generate a genetic map of the mosquito Anopheles gambiae. In the present study we explore the potential of microsatellites as a tool for studying the genetic structure of natural populations of this malaria vector. Genetic polymorphism at twenty enzyme coding gene loci and eleven microsatellite DNA loci was surveyed in a population of An. gambiae from Mali, West Africa. All of the microsatellite loci surveyed were polymorphic, as compared to 40% of the isozyme loci. The mean heterozygosity for the isozyme loci was only 0.097 (+/- 0.0035), but for the microsatellite loci it was 0.732 (+/- 0.060). The pattern of variability was very different between isozymes and microsatellites. Typically, at an isozyme locus a single allele occurred at a frequency > or = 0.75, whereas at microsatellite loci the most common allele had a frequency < 0.50. We conclude that microsatellites provide a rich source of genetic polymorphisms for the study of the population genetics of An. gambiae and are in many ways superior to isozymes for this purpose. We discuss the potential for utilizing genetically mapped microsatellite loci to explore the effect of chromosomal inversions on the distribution of genetic polymorphisms in An. gambiae.

Africa, Western↗

PubMed1995

Soluble CD14 is increased in bronchoalveolar lavage of active sarcoidosis and correlates with alveolar macrophage membrane-bound CD14.

CD14 is a myeloid differentiation antigen which exists in a membrane-bound (55 kD) and a soluble (48 kD) form. This antigen is a receptor for lipopolysaccharide (LPS) structures and triggers the production of various cytokines. The aim of this study was to evaluate whether in active sarcoidosis, a disease with increased proportions of alveolar macrophages (AM) with CD14 expression in BAL fluid, the soluble form of CD14 (sCD14) is also increased. The sCD14 levels were measured in BAL fluid with an ELISA, and membrane-bound CD14 was determined by an immunoperoxidase assay, in active sarcoidosis (n = 13), inactive sarcoidosis (n = 9), idiopathic pulmonary fibrosis (IPF) (n = 6), and control subjects (n = 8). Higher concentrations of sCD14 were present in BAL fluid of patients with active sarcoidosis (58 +/- 34 ng/ml) than in those with inactive disease (13 +/- 10 ng/ml), patients with IPF (5 +/- 5 ng/ml), or control subjects (10 +/- 8% ng/ml) (p < 0.01). Similarly, the proportions of AM expressing membrane-bound CD14 were increased in active sarcoidosis (91 +/- 6%) compared with inactive sarcoidosis (82 +/- 6%), patients with IPF (76 +/- 13%), and control subjects (79 +/- 9%) (p < .05). In sarcoidosis, a significant correlation was found between the sCD14 concentration in BAL fluid and AM membrane expression of CD14 (r = 0.57, p < 0.01). We conclude that sCD14 is increased in BAL of active sarcoidosis suggesting a potential role for this substance as marker of activity and in the pathogenesis of pulmonary sarcoidosis.

Adult↗

PubMed1995

Lymphocyte subsets in bronchoalveolar lavage fluid of children without bronchopulmonary disease.

Bronchoalveolar lavage (BAL) is increasingly used in the pediatric age group. However, normal values for BAL fluid (BALF) constituents are lacking. As part of an investigation to define reference values in children, we studied lymphocyte surface markers of BALF in 28 children 3 to 16 yr of age without bronchopulmonary disease. All of them were undergoing elective surgery for nonpulmonary illnesses. BAL was performed under general anaesthesia with tracheal intubation. A flexible bronchoscope (Pentax 3.5 or 4.9 mm) was wedged into the middle lobe or into one of its segments, and 3 x 1 ml/kg body weight normal saline warmed to body temperature were instilled and immediately withdrawn. The first sample was studied separately; subsequent samples were pooled for analysis. Studies on lymphocyte surface markers were performed on the pooled sample only. The distribution of B-cells, pan T-cells, and CD57 positive cells was within the range reported for adult subjects. However, CD4/CD8 ratios were lower than in adults (0.7 +/- 0.4, mean +/- SD). This decrease in the CD4/CD8 ratio was caused by an increase in CD8 cells. Comparative analysis of blood and BALF lymphocytes in a subgroup of children showed that CD4/CD8 ratios in blood were within the normal range reported for this age group and significantly higher when compared with BALF. The lower CD4/CD8 ratios in normal children have to be considered in the interpretation of BALF in children with pulmonary diseases.

Adolescent↗

PubMed1995

Alveolar macrophage TNF-alpha release and BAL cell phenotypes in sarcoidosis.

The aim of this study was to investigate the relationship between release of tumor necrosis factor-alpha (TNF-alpha) by alveolar macrophages (AM) and the phenotypic characteristics of bronchoalveolar lavage (BAL) cells in sarcoidosis. We studied the spontaneous release of TNF-alpha by AM in vitro and the phenotypic characteristics of freshly recovered BAL T-cells and AM in 31 individuals (13 with active sarcoidosis, nine with inactive sarcoidosis, and nine normal controls). TNF-alpha was measured by enzyme-linked immunosorbent assay (ELISA) in supernatants from unstimulated AM after 24 h culture. Phenotypic markers of BAL cells were determined by an immunocytochemical assay. AM of patients with active sarcoidosis released more TNF-alpha (1,355 +/- 133 pg/ml/ 10(6) AM/24 h) than those of the inactive group (651 +/- 142 pg/ml/10(6) AM/24 h) or the normal controls (425 +/- 121 pg/ml/10(6) AM/24 h), with p < 0.001 for both comparisons. The amount of TNF-alpha released correlated positively with the percentage expression of CD4 (r = 0.72) and CD25 (r = 0.70) by lymphocytes, and of CD14 (r = 0.63), VLA-4 (r = 0.59), FRD1 (r = 0.67) and 27E10 (r = 0.67) by AM, with p < 0.001 for all correlations. In conclusion, this relationship suggests that these antigens may be considered as cellular activation markers, and that some of these AM antigens may indirectly characterize the AM phenotype that is capable of producing TNF-alpha.

Adult↗

PubMed1995

Effects of the phospholipase-C inhibitor, U73122, on signaling and secretion in pituitary gonadotrophs.

The effects of inhibition of phosphoinositide hydrolysis by U73122 [1-(6-[17 beta-3-methoxyestra-1,3,5- (10) triene-17-yl] amino/hexyl) 1H-pyrroledione] and neomycin on agonist-stimulated intracellular signaling and secretory responses were analyzed in cultured pituitary cells and alpha T3-1 gonadotrophs. GnRH (100 nM)- and endothelin-1 (ET-1; 100 nM)-induced inositol (1,4,5)-trisphosphate and diacylglycerol formation in normal cells and immortalized gonadotrophs were reduced by U73122 in a concentration-dependent manner, with IC50 values of about 2 microM and complete inhibition at 10 microM U73122. Neomycin also reduced GnRH- and ET-induced inositol phosphate production in both cell types. Agonist-induced intracellular Ca2+ responses were also inhibited in both cell types by U73122 and neomycin at the same concentrations that inhibited their inositol phosphate responses. In cultured pituitary cells, agonist-induced LH release was inhibited by U73122 and neomycin in a dose-dependent manner. In perifused pituitary cells, U73122 completely inhibited GnRH- and ET-1-induced LH release, but after 10 min caused a progressive and substantial increase in basal LH release. In static cultures, U73122 inhibited agonist-induced LH response at low concentrations (up to 3 microM), but stimulated LH release at higher concentrations due to direct activation of exocytosis by the compound. When added alone, U73122 caused a concentration-dependent increase in LH release with an EC50 of about 7 microM and a maximum response similar that that elicited by GnRH. The stimulatory action of U73122 on LH release was not reduced in the absence of extracellular Ca2+. In contrast to cultured pituitary cells, alpha T3-1 gonadotrophs showed only constitutive exocytosis that was not affected by either neomycin or U73122. These results demonstrate that GnRH and ET(A) receptors are coupled to the phosphoinositide/Ca2+ transduction system in pituitary gonadotrophs, and provide evidence for the dependence of agonist-regulated exocytosis on this signaling pathway. The ability of U73122 to stimulate LH release could reflect an additional action of the compound at late steps in the exocytic pathway.

Animals↗

PubMed1995

Gonadotropin-releasing hormone-induced sensitization of calcium-dependent exocytosis in pituitary gonadotrophs.

Agonist-induced increases in cytoplasmic calcium concentration ([Ca2+]i) play a pivotal role in regulated exocytosis by promoting the fusion of secretory vesicles with the plasma membrane. In permeabilized and ATP-primed pituitary cells, increases in ambient [Ca2+]i stimulated the release of LH from gonadotrophs with an EC50 of 2-3 microM. In contrast, the responses of intact gonadotrophs to agonist stimulation by GnRH were characterized by transient [Ca2+]i elevations of up to 1.5 microM, followed by a plateau of 300-400 nM. The sensitivity of the exocytotic response of permeabilized cells to [Ca2+]i was significantly increased by GnRH, which reduced the EC50 for [Ca2+]i to the submicromolar concentration range. The stimulatory action of GnRH on LH release in permeabilized cells was not a consequence of intracellular Ca2+ release, but was associated with increased cytidine diphosphate diacylglycerol production. Activation of protein kinase C by phorbol esters caused a similar increase in the Ca2+ sensitivity of LH release from permeabilized cells, and this effect was not additive to that of GnRH. Furthermore, the GnRH-induced increase in the sensitivity of the exocytotic response to Ca2+ was attenuated by inhibitors of protein kinase C. These findings indicate that although elevated [Ca2+]i per se can promote LH release from permeabilized gonadotrophs, concomitant activation of protein kinase C is necessary to support exocytosis at the physiological [Ca2+]i levels that prevail in GnRH-stimulated intact cells. Such sensitization of the Ca(2+)-dependent secretory mechanism by protein kinase C may be an important step in the agonist-induced release of LH from pituitary gonadotrophs.

Adenosine Triphosphate↗

PubMed1995

frizzled regulates mirror-symmetric pattern formation in the Drosophila eye.

Coordinated morphogenesis of ommatidia during Drosophila eye development establishes a mirror-image symmetric pattern across the entire eye bisected by an anteroposterior equator. We have investigated the mechanisms by which this pattern formation occurs and our results suggest that morphogenesis is coordinated by a graded signal transmitted bidirectionally from the presumptive equator to the dorsal and ventral poles. This signal is mediated by frizzled, which encodes a cell surface transmembrane protein. Mosaic analysis indicates that frizzled acts non-autonomously in an equatorial to polar direction. It also indicates that relative levels of frizzled in photoreceptor cells R3 and R4 of each ommatidium affect their positional fate choices such that the cell with greater frizzled activity becomes an R3 cell and the cell with less frizzled activity becomes an R4 cell. Moreover, this bias affects the choice an ommatidium makes as to which direction to rotate. Equator-outwards progression of elav expression and expression of the nemo gene in the morphogenetic furrow are regulated by frizzled, which itself is dynamically expressed about the morphogenetic furrow. We propose that frizzled mediates a bidirectional signal emanating from the equator.

Animals↗

PubMed1995

Increased soluble CD14 levels in BAL fluid in pulmonary tuberculosis.

INTRODUCTION: CD14 is a cell surface glycoprotein expressed mainly on mature monocytes and macrophages. Soluble CD14 (sCD14) is present in normal plasma and is found increased in serum of patients with septicemia, polytrauma, and sarcoidosis. In active sarcoidosis, increased levels of sCD14 in BAL supernatant have been demonstrated. STUDY OBJECTIVE: To investigate sCD14 levels in BAL of pulmonary tuberculosis (PTB), another inflammatory disease characterized by granuloma formation. METHODS: BAL was performed in 12 patients with active but untreated PTB and 12 healthy persons. Cytologic and immunocytologic characteristics were determined. sCD14 was measured by a sandwich enzyme-linked immunosorbent assay. RESULTS: The level of sCD14 in patients with PTB was increased compared with controls (mean +/- SEM: 34.4 +/- 9.6 ng/mL vs 11.5 +/- 2.2 ng/mL; p = 0.02). No correlation was found between sCD14 levels and BAL cell differentials or lymphocyte surface markers. CONCLUSION: Similar to sarcoidosis, increased levels of sCD14 are found in BAL supernatant of PTB patients.

Adult↗

PubMed1995

[A study on expression of epidermal growth factor receptor in pleomorphic adenoma of lacrimal gland].

Expression of epidermal growth factor (EGF) receptor was examined in 32 cases of pleomorphic adenoma of lacrimal gland by means of an immunohistochemical method. While normal lacrimal glands were all negative for the antigen, EGF receptor was positive in 10 pleomorphic adenomas and the positive staining was mainly limited in the tumor cells in trabecular or duct-like arrangement or squamous metaplastic epithelium. These data suggest that the expression of EGF receptor be significantly higher in pleomorphic adenoma than in normal lacrimal gland, and also suggest that EGF receptor be expressed in the neoplastic cells which are considered to be of duct origin.

Adenoma, Pleomorphic↗