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Biomedical subjects

L Ding

Publications and source records attributed to L Ding.

At least 91 records · Page 5Linked to original sources
PubMed1999

[Association and isochromatization of ion-pair of the acid and basic].

In this paper, the method for forming the isochromatic dye ion-pair was described, and the mechanism and the types of the association of the ion-pair studied. The isochromatization of the associated reaction and the solvent effect were discussed by the electron theory with the absorption spectrum and energy diagram. The results of the extraction spectrophotometric method of the isochromatic dye ion-pairs were shown in a table, indicating the high sensitivity of the isochromatic dye ion-pairs. It is proven that the method of the extraction spectrophotometry, the flotation spectrophotometry and the extraction fluorometry was of high and superhigh sensitivity.

English Abstract↗
PubMed1999

[Absorption characteristics of the isochromatic dye ion-pair formed by tetrabromofluorescein and buthylrhodamine B].

Absorption properties of tetrabromofluorescein (TBF) and buthylrhodamine B (BRB) and the isochromatic dye ion-pair formed by TBF and BRB were studied. The experiment showed that the proportion of TBF: BRB was 1:2 at pH=7 with Ksp = 2.16 x 10(-18), and the extraction rate was 84.2%. This provides a basis for the application of TBF and BRB to extraction spectrophotometry.

English Abstract↗
PubMed1999

[Determination of diclofenac sodium and codeine phosphate in shuerfen tablets by CPA-matrix method].

The contents of diclofenac sodium and codeine phosphate in shuerfen tablets may be determined simultaneously by CPA-matrix method without separation of its components. The program was edited by BASIC. The average recoveries and RSD were 99.85%, 0.46% for diclofenac sodium and 99.80%, 0.50% codeine phosphate, respectively. This method is simple and rapid, and the result is accurate.

Codeine↗
PubMed1998

Differential effects of CD28 engagement and IL-12 on T cell activation by altered peptide ligands.

To futher our understanding of the mechanisms underlying the diverse effects of altered peptide ligands (APL) on T cell activation, we used a population of nonactivated spleen cells from mice that expressed a transgenic TCR specific for myelin basic protein Ac1-11 and peptide analogues that display either enhanced or decreased affinities for TCR/MHC to address the question whether APL-induced signaling through the TCR can regulate the capability of APC to activate T cells. We demonstrate that weak agonists APL are poor inducers of all aspects of the activation of both the responder T cells and the APC. Enhancement of the antigenic signal by augmenting the binding of the weak agonists to MHC reversed their defective activating capacity. Enhancement of costimulation by engagement of CD28 only resulted in augmentation of the capacity of the weak agonist APL to induce proliferation and IL-2/IL-3 production, but not CD40L or IL-12Rbeta2 chain expression on T cells, CD80/CD86 expression on APC, IL-12 secretion, or IFN-gamma production. Exogenous IL-12 promoted IFN-gamma production in the presence of the weak agonists. These studies demonstrate that there is a critical threshold of antigenic signal required for full activation of the T cell-APC interactions needed for the differentiation of Th1 cells. The provision of excess costimulation can overcome some of the defects in T cell activation by weak agonists, but is insufficient to induce a sufficient level of CD40L expression needed for engagement of CD40 on APC with subsequent IL-12 production and induction of IL-12Rbeta2 chain expression.

Amino Acid Substitution↗
PubMed1998

A selenium-containing abzyme, the activity of which surpassed the level of native glutathione peroxidase.

Using two different glutathione derivatives as hapten, we have prepared two abzymes, which display glutathione peroxidase (GPX) activity. Their GPX activities are 0.2 and 1.6 times that of natural GPX from rabbit liver, respectively. Selenium content analysis indicates that the activity difference between the two abzymes is possibly attributed to the conformation difference of the abzymes.

Animals↗
PubMed1998

WD-40 repeat region regulates Apaf-1 self-association and procaspase-9 activation.

The casp9 protein plays a critical role in apoptosis induced by a variety of death stimuli. A regulator of apoptosis, Apaf-1, binds to and activates pro-casp9 in the presence of cytochrome c and dATP, a requirement that is bypassed by deletion of the WD-40 repeats located in the C-terminal half of Apaf-1. In this report, we used constitutively active Apaf-1 mutant lacking the WD-40 repeat region to study the mechanism and regulation of pro-casp9 activation. Mutational analysis revealed that only a small portion of the CED-4 homologous region (residues 456-559) could be deleted without destroying the ability of Apaf-1-(1-559) to activate pro-casp9. Apaf-1 can self-associate to form oligomers. Disruption of Apaf-1 self-association by deletion (Delta109-559) or mutation of the P-loop region (K149R) abrogated Apaf-1-mediated pro-casp9 activation. Forced oligomerization of the caspase recruitment domain of Apaf-1 was sufficient for pro-casp9 activation. Dimerization of chimeric Fpk-pro-casp9 protein with the dimerizer drug FK1012 induced pro-casp9 processing and apoptosis in cells. Significantly, the C-terminal region containing WD-40 repeats interacted with its N-terminal CED-4 homologous region, as determined by immunoprecipitation experiments. Importantly, expression of the WD-40 repeat region inhibited Apaf-1 self-association and proteolytic activation of pro-casp9. These studies provide a mechanism by which Apaf-1 promotes autoactivation of pro-casp9 through Apaf-1 self-association, a process that is negatively regulated by the WD-40 repeats.

Animals↗
PubMed1998

The cloning and characterization of a novel human diacylglycerol kinase, DGKiota.

Diacylglycerol (DAG) plays a central role in both the synthesis of complex lipids and in intracellular signaling; diacylglycerol kinase (DGK) catalyzes the phosphorylation of DAG, which yields phosphatidic acid. A family of DGKs has been identified in multicellular organisms over the past few years, but the physiological function(s) of this diversity is not clear. One clue has come from the Drosophila DGK2, rdgA, since mutations in this gene cause retinal degeneration. We isolated a novel DGK, which we designated DGKiota, from human retina and brain libraries. DGKiota contains two cysteine-rich repeats, a region similar to the phosphorylation site domain of myristoylated alanine-rich C kinase substrate, a conserved catalytic domain, and four ankyrin repeats at its C terminus. By primary structure, it is most similar to human DGKzeta and Drosophila rdgA. An >12-kilobase mRNA for DGKiota was detected only in brain and retina among the tissues examined. In cells transfected with the DGKiota cDNA, we detected an approximately 130-kDa protein by immunoassay, and activity assays demonstrated that it encodes a functional DAG kinase. The protein was found to be in both the cytoplasm and nucleus with the localization controlled by PKC isoforms alpha and gamma. The gene encoding DGKiota was localized to human chromosome 7q32.3-33, which is known to be a locus for an inherited form of retinitis pigmentosa. These results have defined a novel isoform of DAG kinase, which may have important cellular functions in the retina and brain.

Amino Acid Sequence↗
PubMed1998

The cloning and developmental regulation of murine diacylglycerol kinase zeta.

Diacylglycerol kinases (DGKs) regulate the key signaling intermediates diacylglycerol (DAG) and phosphatidic acid (PA). We isolated cDNA clones of mouse diacylglycerol kinase zeta (mDGKzeta) and found that it shares 88% identity at the nucleic acid level and 95.5% identity at the amino acid level with human DGKzeta (hDGKzeta). Murine DGKzeta protein rose gradually during embryonic development, and was abundant in newborn and adult brains. By RNA whole-mount in situ hybridization, mDGKzeta was shown to be expressed in spinal ganglia and limb buds at low level in E11.5 embryos and at higher level in E12.5 embryos. In E13.5 embryos, DGKzeta mRNA was highly expressed in vibrissa follicles, in spinal ganglia, and in the interdigital regions of the developing limbs. Northern blotting showed that DGKzeta expression was limited to specific anatomical regions of the brain. Thus, the expression of DGKzeta is regulated temporally and spatially during mammalian development and correlates with the development of sensory neurons and regions undergoing apoptosis.

Amino Acid Sequence↗
PubMed1998

Biochemical characterization of selenium-containing catalytic antibody as a cytosolic glutathione peroxidase mimic.

A selenium-containing catalytic antibody (Se-4A4), prepared by converting reactive serine residues of a monoclonal antibody (4A4) raised against a GSH derivative into selenocysteines, acts as a mimic of cytosolic glutathione peroxidase (cGPX). To clarify the mechanism of action of this catalytic antibody, detailed studies on kinetic behaviour and biological activity were carried out. A rate of acceleration (kcat/Km/kuncat) 10(7)-fold that of the uncatalytic reaction is observed. Under similar conditions, the turnover number (kcat) of Se-4A4 is 42% of that of the natural rabbit liver cGPX. The Se-4A4 reaction involves a Ping Pong mechanism, which is the same as that of the natural cGPX. The selenocysteine residue is located in the binding site of the antibody and is shown to be crucial for this activity. Of the thiol compounds tested, only GSH is able to serve as substrate for Se-4A4. It was demonstrated, using the free-radical-damage system (hypoxanthine/xanthine oxidase) of cardiac mitochondria, that Se-4A4 can protect mitochondria from free-radical damage at least 10(4)-fold more effectively than the natural cGPX.

Antibodies, Catalytic↗
PubMed1998

Analysis of water from the Space Shuttle and Mir Space Station by ion chromatography and capillary electrophoresis.

Drinking water and condensate samples collected from the US Space Shuttle and the Russian Mir Space Station are analyzed routinely at the NASA-Johnson Space Center as part of an ongoing effort to verify water quality and monitor the environment of the spacecraft. Water quality monitoring is particularly important for the Mir water supply because approximately half of the water consumed is recovered from humidity condensate. Drinking water on Shuttle is derived from the fuel cells. Because there is little equipment on board the spacecraft for monitoring the water quality, samples collected by the crew are transported to Earth on Shuttle or Soyuz vehicles, and analyzed exhaustively. As part of the test battery, anions and cations are measured by ion chromatography, and carboxylates and amines by capillary electrophoresis. Analytical data from Shuttle water samples collected before and after several missions, and Mir condensate and potable recovered water samples representing several recent missions are presented and discussed. Results show that Shuttle water is of distilled quality, and Mir recovered water contains various levels of minerals imparted during the recovery processes as designed. Organic ions are rarely detected in potable water samples, but were present in humidity condensate samples.

Chromatography, Ion Exchange↗
PubMed1998

[Polymorphisms of four microsatellite markers closely linked with PKD1 in Chinese].

OBJECTIVE: To investigate the polymorphisms and evaluate the value in gene diagnosis of four microsatellites, which are the KG8 that locating in 3'untranslated region of PKD1 and SM6, CW4 and CW2, closely linked to PKD1. METHODS: The DNAs of some unrelated Chinese Han people were analyzed by using PCR, PAGE and silver staining and the findings were compared with those reported on Caucasian. RESULTS: In Chinese, KG8 included 6 alleles and PIC was 0.312; in Caucasian,8 alleles,PIC 0.545. The number of alleles and PIC of KG8 in Chinese were not so many as those in Caucasian. SM6 included 24 kinds of alleles and PIC was 0.80 in Chinese; in Caucasian,16 alleles, PIC 0. 653. CW4 is 9 alleles in Chinese and PIC of SM6 were more than those in Caucasian. In Chinese CW4, 9 alleles, PIC 0.850; in Caucasian, 9 alleles, PIC 0.782. Although the number of CW4 allele in the two nations was the same, PIC in Chinese was higher than that in Caucasian. In Chinese CW2,7 alleles, PIC 0.814; and in Caucasian, 13 alleles, PIC 0.809. The number of allele of CW2 in Chinese was not so many as that in Caucasian; however,the PIC was about the same. CONCLUSION: The results indicate that there are differences in the number of allele and distribution of the four microsatellite markers between Chinese and Caucasian. So there exists difference of dinucleotide repeat between two nations. In addition,SM6, CW4 and CW2 could be used as valuable markers in gene linkage diagnosis and personal and related identification in forensic medicine.

Alleles↗
PubMed1998

Inactivation of a common epitope responsible for the induction of antibody-dependent enhancement of HIV.

BACKGROUND: The primary antigenic domain responsible for complement-mediated antibody-dependent enhancement (C'-ADE) of HIV and simian immunodeficiency virus resides in the principal immunodominant sequence of the transmembrane protein. OBJECTIVE: To identify whether there are amino-acid residues common to the epitopes of the known enhancing human monoclonal antibodies (MAb), and to provide a structural model for this functional region present on the HIV envelope. Since our model predicts that this region is involved in the association of gp120 with gp41, this association was monitored for each mutant. DESIGN: The binding of enhancing human MAb to point and deletion mutations within the enhancing domain was analyzed by two methods. The first analyzed binding to mutants expressed in COS cells: the second quantified the binding of four enhancing human MAb to each mutant gp160 versus wild-type control by enzyme-linked immunosorbent assay (ELISA). METHODS: Site-directed mutagenesis was used to produce specific deletions and point mutants, which were expressed in COS cells. Binding of MAb 50-69 and V3-loop MAb 5F7 were visualized in the wild-type and each of the mutant constructs by immunohistochemistry. Quantitative evaluation of enhancing human MAb binding to each mutant versus wild-type was performed by ELISA. A model for the enhancing domain and its relationship to gp120 association with gp41 was provided by molecular dynamics and ligand docking methods. RESULTS: All available enhancing human MAb known to bind to the principal immunodominant region of gp41 were unable to bind to deletions involving the disulfide loop, which in our molecular model provided the primary association site between gp120 and gp41. Point mutations in the loop blocked this association, but had a quantitatively smaller effect on the binding of the enhancing human MAb. A conservative W596Y mutation completely blocked the binding of all human MAb, but had no effect on gp120-gp41 association. CONCLUSIONS: A variety of mutations within the primary C'-ADE domain inhibit binding of enhancing human MAb as well as blocking the association of gp120 and gp41. A conservative W596Y mutation blocks binding of all enhancing human MAb with retention of gp120-gp41 association. These data are important to the design of vaccines in which the primary enhancing epitope is disarmed to prevent the subsequent induction of an amnestic response that could lead to viral enhancement of infection. The retention of the gp120-gp41 association is postulated to yield an immunogen similar to natural infection for both subunit and genetic vaccines.

Amino Acid Sequence↗
PubMed1998

Immunogenicity and antitumor activity of a liposomal MUC1 peptide-based vaccine.

A human MUC1-transfected mouse mammary adenocarcinoma cell line (GZHI) was used to develop both subcutaneous and intravenous tumor models. A vaccine formulation comprised of a 24 mer (human MUC1) synthetic peptide encapsulated with monophosphoryl lipid A adjuvant (MPLA) in multilamellar liposomes was tested for immunogenicity and anti-tumor activity. A low dose of the human MUC1 peptide (5 microg) administered in liposomes provided excellent protection of mice in both tumor challenge models. The protective antitumor activity mediated by the liposome formulation correlated with anti-MUC1-specific T-cell proliferation, gamma-interferon (IFN-gamma) production and IgG2a anti-MUC1 antibodies, suggesting a type 1 (T1) T-cell response. In contrast, lack of protection in mice immunized with negative control vaccines correlated with IgG1 anti-MUCI antibody formation, low or no anti-MUC1 IgG2a and low antigen-specific T-cell proliferation, consistent with a type 2 (T2) T-cell response to the tumor.

Amino Acid Sequence↗
PubMed1998

Pancreatic microcirculation in acute pancreatitis.

We present a review of the microvascular morphology of the pancreas and microstructure of the pancreatic lobule, and report our experimental results of the investigation of pancreatic microcirculation following acute pancreatitis. Impairment of pancreatic microcirculation in the early phase of acute pancreatitis may play a key role in the progression of this disease. Possible contributory mechanisms include increased vascular permeability, reduced blood flow, leukocyte-endothelial cell interaction and intravascular thrombus formation. Using an in-vivo microscope system and off-line computer analysis, we achieved direct visualization and quantification of changes in microvascular permeability and leukocyte behavior in pancreas with acute pancreatitis. Bradykinin and oxygen radicals have been demonstrated to be involved in the increase of vascular permeability in the early stage of caerulein pancreatitis. Leukocyte adherence to the vessels in the pancreatic microcirculation is a secondary event following permeability changes in acute pancreatitis. Leukocyte infiltration during exacerbation of acute pancreatitis is mediated by leukocyte-endothelial cell interaction via leukocyte integrin CD11b/18.

Acute Disease↗
PubMed1998

Vertical transmission of human immunodeficiency virus type 1: frequency and correlation of transmission (review).

Child bearing age women constitute the fastest growing subset of the human immunodeficiency virus (HIV) infected population. Consistent with this fact is the increasing number of HIV infected children. Vertical transmission from mother to child is the most common route of infection. Because of the nature of the virus, it is difficult to determine the timing of infection. There are three time periods during which infection can occur: prenatally, perinatally, or postnatally. Evidence for each is presented within this review.

Breast Feeding↗
PubMed1998

Infection with the human immunodeficiency virus type 2: epidemiology and transmission (Review).

Human immunodeficiency virus type 2 (HIV-2) is known to be one of the agents that causes acquired immunodeficiency syndrome (AIDS). It has been present in West Africa since the 1960s and is currently epidemic there. Compared with human immunodeficiency virus type 1 (HIV-1), HIV-2 is genomically different. Furthermore, it is less prevalent worldwide than HIV-1. In West Africa, seropositive rates of HIV-2 are higher in urban versus rural communities, however, there are no gender differences. Sexual contact and vertical transmission are known modes of infectivity, though HIV-2 is less contagious than HIV-1.

Africa↗
PubMed1998

Cisplatin and phorbol ester independently induce ERCC-1 protein in human ovarian carcinoma cells.

Nucleotide excision repair (NER) is the DNA repair pathway by which cisplatin-induced damage is removed from DNA in human cells. ERCC-1 is one of the essential proteins in NER, and is essential for life. Enhanced ERCC-1 expression has been associated with clinical and cellular resistance to cisplatin. We therefore carried out this study to investigate the effect of cisplatin on ERCC-1 protein expression in A2780/CP70 human ovarian cancer cells. Western blot analysis showed that ERCC-1 protein levels were increased to more than 3 times control after a 1 h cisplatin exposure to A2780/CP70 cells in culture. This increase was time- and concentration-dependent. The effect of cisplatin was maximal at 40 mM and peaked 24-48 h after exposure to the drug. These results extend our previous observations that ERCC-1 mRNA expression is induced by cisplatin in this system. TPA, a known AP-1 activator and tumor-promoting phorbol ester, also induced ERCC-1 protein to the same extent as cisplatin, but did not synergize with cisplatin in this regard. These findings suggest that ERCC-1 gene up-regulation in these cells can result through a DNA damage-response pathway, or through the induction of AP-1 activity, independent of the occurrence of DNA damage.

Antineoplastic Agents↗