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Biomedical subjects

Q Li

Publications and source records attributed to Q Li.

At least 631 records · Page 35Linked to original sources
PubMed1997

Apoptosis in patients with posterior uveitis.

BACKGROUND: Apoptosis plays a part in the pathogenesis of autoimmune diseases. OBJECTIVE: To investigate the expression of apoptotic markers in the eyes of patients with uveitis. METHODS: With the use of immunohistochemical and in situ apoptotic detection techniques, apoptotic molecules (Fas or Fas ligand [FasL]) and nuclear DNA fragmentation were examined in 8 enucleated eyes with Behçet's disease (1), sarcoidosis (1), subretinal fibrosis and uveitis (1), sympathetic ophthalmia (4), and the Vogt-Koyanagi-Harada syndrome (1); in 5 chorioretinal biopsy specimens with acute retinal necrosis (2), multifocal choroiditis (1), sarcoidosis (1), and subretinal fibrosis and uveitis (1); and in 3 normal control eyes. RESULTS: Fas and FasL were constitutively expressed in the normal human retina, but they were expressed much less in the choroid. Increased expression of Fas and FasL was found in the retina, chorioretinal scar, and choroidal granulomas in uveitic eyes. However, Fas and FasL expression was absent in the biopsy specimens with acute retinal necrosis, and little Fas or FasL was noted on infiltrating lymphocytes. DNA fragmentation was also identified in eyes with chorioretinal scar and gliosis. CONCLUSIONS: Apoptosis occurs in uveitic eyes and may play a regulatory role in limiting ocular inflammation. In uveitic eyes, a dysregulation of the Fas-FasL apoptotic pathway may lead to gliosis and fibrosis.

Adolescent↗
PubMed1997

Mutagenesis screens for interacting genes reveal three roles for fat facets during Drosophila eye development.

The Drosophila fat facets gene encodes a deubiquitinating enzyme that regulates a cell communication pathway essential early during eye development to inhibit the determination of excess photoreceptors. Ubiquitin is a small polypeptide that tags proteins for degradation by a multisubunit proteolytic complex called the proteasome. The FAT FACETS protein is thought to be required to remove ubiquitin from a particular protein, thereby rescuing if from proteolysis. In order to identify the genes encoding the substrate of FAT FACETS and other components of the neural inhibition pathway, a mutagenesis screen for dominant enhancers of the fat facets mutant eye phenotype was performed. Several genes were identified, one of which is an excellent candidate for encoding a component of the pathway regulated by FAT FACETS. Three different eye phenotypes were observed when the fat facets mutants were dominantly enhanced by different mutations, suggesting that fat facets has other functions in addition to its critical role early in eye development.

Animals↗
PubMed1997

Comparison of swallowing function in Parkinson's disease and progressive supranuclear palsy.

Dysphagia is common in both Parkinson's disease (PD) and progressive supranuclear palsy (PSP). Although it is believed to be more common in PSP, there are no controlled data and no comparison of swallowing function between these two disorders. Our aim was to assess dysphagia and swallow function in patients with PSP and PD. Seven patients with PSP were matched to seven patients with PD on the basis of disease duration. Self-rated dysphagia, movement disorder disability, modified barium swallow results, and abnormalities noted on manometry of the lower esophageal sphincter, esophageal body, upper esophageal sphincter, and pharynx were compared between the two groups. Neither severity nor duration of dysphagia differed between the two groups. Patients with PSP had a significantly greater degree of disability [median (range) Hoehn & Yahr score, 4 (3-5) vs. 2 (1-2); P < 0.002]. Manometric abnormalities were similar for the two groups. Oral-phase abnormalities on modified barium swallow were significantly more frequent in PSP (four patients with PSP vs. no patients with PD; p < 0.005). Pharyngeal abnormalities did not differ. Modified barium-swallow scores correlated well with self-reported dysphagia severity for patients with PSP (r = 0.93; p < 0.05) but not for those with PD (r = 0.42; p = NS). The frequency of abnormalities noted during the oral phase was significantly increased in PSP. It is hypothesized that the sensory information conveyed due to this may account for the better correlation between symptoms and swallowing abnormalities and the belief that swallowing problems are more common in PSP.

Barium↗
PubMed1997

Serum-free culture conditions for cells capable of producing long-term survival in lethally irradiated mice.

The goal of ex vivo culture is to expand and/or differentiate cells in culture such that they retain their functional characteristics when reinfused into a patient. The studies presented here analyzed the use of culture conditions devoid of serum to expand murine hematopoietic stem cells. Bone marrow cells from male B6D2F1/J mice were cultured for up to 28 days in serum-free medium in the absence or presence of stem cell factor (SCF), GM-CSF or a combination of the two factors. Cells cultured for up to 21 days were assessed for granulocyte-macrophage colony-forming cells (GM-CFC), spleen colony-forming units, and cells responsible for short-term and long-term hematopoietic repopulation in lethally irradiated mice. Compared to initial seeding levels, the presence of SCF and GM-CSF increased total cell numbers 90-fold and GM-CFC numbers 42-fold over a 21-28 day culture period. Although spleen colony-forming unit cells did not increase, they were maintained at initial seeding levels over a 21-day period in the presence of SCF and GM-CSF. In lethally irradiated mice, survival enhancement and hematologic reconstitution were optimum with cells cultured for only seven days: survival at six months was 100% with cells cultured in SCF plus GM-CSF or SCF alone, compared to 50% with cells cultured with only GM-CSF. Hybridization analysis of bone marrow, spleen and thymus DNA from irradiated mice transplanted with these cultured cells confirmed male donor cell-derived repopulation at 45 days and 180 days post-transplant. These studies illustrate that murine GM-CFC can be expanded and that long-term repopulating hematopoietic cells can, at the minimum, be maintained ex vivo in serum-free culture. The use of defined serum-free culture systems holds great promise for further evaluation of the mechanisms that control hematopoietic stem cell proliferation.

Animals↗
PubMed1997

Fuchs' corneal endothelial cells transduced with the human papilloma virus E6/E7 oncogenes.

We sought to develop corneal endothelial cell cultures with extended lifespan from corneas with Fuchs' dystrophy. Descemet's-endothelial cell explants from histology confirmed recipient corneas of two patients with Fuchs' dystrophy were cultured. After a small number of corneal endothelial cells with irregular, endothelial morphology migrated from the explants onto the culture plate, the cells were transduced with a disabled retrovirus (pLXSN16E6/E7) coding for the human papilloma virus type 16 transforming oncoproteins E6 and E7. Expression of E6/E7 mRNA in the cell cultures with extended lifespan was monitored by RT-PCR. In vitro labeled cellular protein patterns of the Fuchs' E6/E7 cell cultures with extended lifespan were compared with those from normal human corneal endothelial E6/E7 cell cultures with extended lifespan using two-dimensional gel electrophoresis. Two endothelial cell cultures with extended lifespan were derived from Fuchs' corneas. The morphology of the Fuchs' cells expressing E6 and E7 was similar to that of normal corneal endothelial cells expressing E6 and E7. Surprisingly, the rate of proliferation of Fuchs'-derived cells was similar to that of normal endothelial cells transduced with E6/E7. Proliferation of each Fuchs' cell culture with extended lifespan continued for over 30 population doublings. There were limited quantitative differences in the two-dimensional gel electrophoretic protein patterns of the Fuchs'-derived and normal endothelial cell cultures with extended lifespan, respectively. Retroviral integration is dependent on cell proliferation. Thus, cells that migrated from the Fuchs' Descemet's explants were undergoing at least limited in vitro proliferation when the retroviral vector coding for E6/E7 integrated. Fuchs' corneal endothelial cells expressing E6 and E7 had similar proliferation, cellular morphology, and two-dimensional gel protein electrophoretic patterns to normal corneal endothelial cells expressing the same oncoproteins.

Cell Division↗
PubMed1997

Differential inhibition of DNA synthesis in human T cells by the cigarette tar components hydroquinone and catechol.

Hydroquinone (HQ), catechol, and phenol exist in microgram quantities in cigarette tar and represent the predominant form of human exposure to benzene. Exposure of human T lymphoblasts (HTL) in vitro to 50 microM HQ or 50 microM catechol decreased IL-2-dependent DNA synthesis and cell proliferation by >90% with no effect on cell viability. Phenol had no effect on HTL proliferation at concentrations up to 1 mm. The addition of HQ or catechol to proliferating HTL blocked 3H-TdR uptake by >90% within 2 hr without significantly affecting 3H-UR uptake, suggesting that both compounds inhibit a rate-limiting step in DNA synthesis. However, the effects of HQ and catechol appear to involve different mechanisms. Ferric chloride (FeCl3) reversed the inhibitory effect of catechol, but not HQ, corresponding with the known ability of catechol to chelate iron. HQ, but not catechol, caused a decrease in transferrin receptor (TfR, CD71) expression, comparable to the level observed in IL-2-starved cells. HQ also inhibited DNA synthesis in cultures of transformed Jurkat T lymphocytes, primary and transformed fibroblasts, and mink lung epithelial cells, indicating that its antiproliferative effect was not restricted to IL-2 mediated proliferation. However, DNA synthesis by primary lymphocytes was more sensitive to HQ (IC50 = 6 microM) than that of the transformed Jurkat T cell line (IC50 = 37 microM) or primary human fibroblasts (IC50 = 45 microM), suggesting that normal lymphocytes may be particularly sensitive to HQ. The effects of HQ and catechol on DNA synthesis could be partially reversed by a combination of adenosine deoxyribose and guanosine deoxyribose, suggesting that both compounds may inhibit ribonucleotide reductase.

Animals↗
PubMed1997

Assays for transcription factors access to nucleosomal DNA.

Several promoters have been shown to have sequence specifically positioned nucleosomes that determine the architecture of the promoter. DNA binding proteins that regulate gene expression are in many cases known to bind to their cognate DNA segments organized within such positioned nucleosomes. It has become increasingly evident that the cooperation of chromatin and transcription factors results in an efficient and fine-tuned regulation of transcription. The first step in a gene induction event must be the access of transcription factors for the regulatory promoter/enhancer target sites. In this perspective it becomes interesting to evaluate the affinity of DNA binding proteins for their cognate binding site in a nucleosome context. Here we describe the preparation of nucleosome probe, a method for in vitro nucleosome reconstitution by salt dilution, purification of the reconstituted mononucleosomes, and characterization of the translational and rotational positions of the nucleosomal DNA. In addition, methods for affinity determination and characterization of protein-nucleosomal DNA interaction, such as methylation protection and methylation interference by dimethyl sulfate, quantitative DNase I footprinting, and electrophoretic mobility shift assay, are described.

Binding, Competitive↗
PubMed1997

Stereoselectivity of actions of the calcium sensitizer [+]-EMD 60263 and its enantiomer [-]-EMD 60264.

The thiadiazinone derivative [+]-EMD 60263 ((+)-5-(l-(alpha-ethylimino-3,4-dimethoxybenzyl)-1,2,3,4-tetrah ydroquinoline -6-yl)-6-methyl-3,6-dihydro-2H-1,3,4 -thiadiazine-2-on) is a Ca(2+)-sensitizing agent with only minor phosphodiesterase inhibitory activity. Our aim was to characterize the inotropic and electrophysiological effects of [+]-EMD 60263 and its enantiomer [-]-EMD 60264 in several cardiac muscle preparations. The Ca(2+)-sensitizing activity resided in the [+]-enantiomer only. [+]-EMD 60263 (3 microM) shifted the EC50 of Ca2+ for contractile activation of skinned fibers of pig heart from 2.41 microM to 0.73 microM, whereas [-]-EMD 60264 (30 microM) was ineffective. In Langendorff-perfused guinea pig hearts, [+]-EMD 60263 and [-]-EMD 60264 induced concentration-dependent positive and negative inotropic effects, respectively; both enantiomers reduced spontaneous heart rate but did not influence perfusion pressure. The maximum increase in force of human atrial trabeculae was 35% of pre-drug control with [+]-EMD 60263 in comparison to 113% with forskolin. In guinea-pig papillary muscles, [+]-EMD 60263 and [-]-EMD 60264 had opposite inotropic responses, however, both agents similarly prolonged action potential duration. Both enantiomers concentration-dependently blocked the rapidly activating component IKr of the delayed rectifier in guinea-pig myocytes. The block saturated at potentials positive to +30 mV, closely resembling the effects of the antiarrhythmic agent E-4031 which had been originally used to define IKr.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1997

Interferon-gamma induced increases in intracellular cathepsin B activity in THP-1 cells are dependent on RNA transcription.

Interferon-gamma (IFN-gamma) treatment of human macrophages induces increased intracellular levels of cathepsin B (CB), a lysosomal cysteine proteinase which is implicated in inflammatory tissue injury. To determine the mechanism of the increase, we studied the macrophage-like cell line, THP-1. Dose and time dependent increases in intracellular CB were seen when cells primed with phorbol ester (PMA) were cultured with IFN-gamma. To determine whether protein synthesis was required for the increase, PMA primed cells were cultured in the presence of IFN-gamma and cycloheximide: The expected increase was inhibited. To determine whether RNA synthesis was required for the IFN-gamma induced increases, PMA primed cells were cultured in the presence of IFN-gamma and actinomycin D. Again the expected increases were not seen. Direct measurement of CB mRNA levels showed increases in cells not treated with inhibitors. These results suggest that the IFN-gamma induced increases in THP-1 cell CB are dependent on RNA and protein synthesis.

Cathepsin B↗
PubMed1997

Production of transgenic mice with yeast artificial chromosomes.

Techniques are now available that allow the transfer of intact yeast artificial chromosome (YAC) DNA into transgenic mice. Coupled with the ability to perform mutagenesis on YAC sequences by homologous recombination in yeast, they enable the analysis of large genes or multigenic loci in vivo. This system has been used to study the developmental regulation of the human beta-globin locus.

Animals↗
PubMed1997

Activation of PKC, superoxide anion production and LDL lipid peroxidation are not dependent on phosphoinositide-specific phospholipase C activity in U937 cells.

Our previous studies have shown that both increase in Ca2+ levels and activation of protein kinase C (PKC) are required for monocyte-mediated O2- production and low density lipoprotein (LDL) peroxidation. Phosphoinositide-specific phospholipase C (phosphoinositidase C or PIC) is believed to mediate release of intracellular Ca2+ through InsP3 formation and activation of PKC through diacylglycerol (DAG). In these studies, we investigated the PIC pathway for its participation in monocytic cell-mediated lipid peroxidation of LDL. We found substantial InsP3 formation in opsonized zymosan (ZOP)-activated U937-b cells, indicating the activation of PIC. Both inhibition of PIC by the PIC inhibitor U-73122 and reduction of the supply of the precursor lipid by lithium chloride suppressed InsP3 formation but did not alter LDL lipid peroxidation nor O2- production by activated cells. Furthermore, we also found that suppression of PIC activity had no substantial inhibitory effect on PKC activity in ZOP-activated human monocytes. Our data suggest that PIC activity is induced upon cell activation resulting in increased levels of InsP3. The activity of this pathway, however, is not required for cell-mediated O2- production, PKC activation or LDL oxidation.

Antioxidants↗
PubMed1997

Application of SSNTDs in radiobiological investigations aboard recoverable satellites.

In recent years some Biostack experiments including a wide spectrum of biological objects have been devoted to study of the radiobiological effects on dry seeds aboard recoverable satellites. Some impressive phenomena have been observed. Clearly, the large amount of energy deposited by the highly ionizing heavy nuclei of cosmic rays is the principal reason for the induced aberrations of the chromosomes of wheat root tip cells. A methodical description of the experimental arrangement and procedure of handling and evaluation of given. The preliminary physical and biological results from the experimental "wheat seeds" are presented.

Chromosome Aberrations↗
PubMed1997

The role of chromatin in transcriptional regulation.

Transcriptional activation is mediated by the facilitated binding of the basal transcription complex to the transcription start site of a promoter. The activation procedure involves protein-protein interactions between specific transcription factors and members of the basal transcription complex. However, since eukaryotic DNA is packaged with histones into nucleosomes the accessibility of the transcription factors is limited. In order to activate transcription, some of the specific transcription factors must have the capacity to bind to their binding sites when organized into nucleosomes. As a next step, the chromatin structure of the promoter needs to be decondensed in order to facilitate the binding of the basal transcription machinery. Recent data have addressed these issues and both binding of transcription factors to their chromatin binding site as well as transcription factor-induced chromatin remodelling have been demonstrated. In addition, factors that are candidates to mediate the chromatin remodelling have recently been identified and characterized. The ability of a transcription factor to recognize its cognate element in a nucleosome is an inheret property that differs among different transcription factors. The implications of the rotational and translational positioning of the DNA within a nucleosome on the accessibility of a transcription factor is described in this review. In addition, nucleosome rearrangement and juxtaposing in the context of transcriptional activation is also discussed.

Animals↗
PubMed1997

Fidelity in functional coupling of the rat P2Y1 receptor to phospholipase C.

1. The rat homologue of the P2Y1 receptor has been heterologously expressed in 1321N1 human astrocytoma cells and in C6 rat glioma cells. 2. As has been shown previously for the turkey and human P2Y1 receptors, the rat P2Y1 receptor expressed in either cell type responded to 2MeSATP with increases in inositol phosphate accumulation that were competitively blocked by the antagonist PPADS. Neither of the wild type cell lines exhibited inositol phosphate responses to P2Y1 receptor agonists. 3. Expression of the rat P2Y1 receptor did not confer a capacity of 2MeSATP to inhibit adenylyl cyclase activity in 1321N1 cells. Moreover, the inhibition of adenylyl cyclase mediated by an endogenous P2Y receptor of C6 glioma cells was not enhanced by expression of the rat P2Y1 receptor. The P2Y receptor-mediated inhibition of adenylyl cyclase in C6 glioma cells expressing both the endogenous P2Y receptor and the rat P2Y1 receptor remained unaffected by PPADS. 4. Since the P2Y receptor responsible for inhibition of adenylyl cyclase in C6 glioma cells does not share the pharmacological or functional properties of the P2Y1 receptor, even when both receptors originate from the same species and are simultaneously expressed in the same cell line, it is concluded that the P2Y1 receptor is distinct from an endogenous P2Y receptor in C6 cells that couples to inhibition of adenylyl cyclase.

Adenosine Triphosphate↗
PubMed1997

Adsorptive stripping voltammetry of bleomycin.

In 0.05 M H2SO4 solution, two reductive peaks, P1 and P2, of bleomycin were obtained. The peak potentials E(P1) and E(P2) were -0.83 and -1.09 V (versus Ag/AgCl), respectively. The sensitivity of P2 was much higher than that of P1. The peak current of P2 was proportional to the concentration of bleomycin over the range 1.0 x 10(-9)-1.0 x 10(-7) M with a detection limit of 5.0 x 10(-10) M using adsorptive voltammetry at an accumulation time of 120 s (Ei = -0.80 V). The behaviour of the reduction wave was studied and applied to the determination of bleomycin in mouse serum. The reduction process of P1 was irreversible with adsorptive characteristics and the adsorption behavior obeyed the Frumkin adsorptive isotherm. The adsorptive coefficient beta was 8.9 x 10(5), the interaction factor alpha was 0.94 and the Gibbs energy of adsorption delta G degree was -33.93 kJ mol-1. P2 was an irreversible adsorption peak with catalytic hydrogen properties.

Adsorption↗