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D Liu

Publications and source records attributed to D Liu.

At least 523 records · Page 29Linked to original sources
PubMed1996

Binding of CD44 to hyaluronic acid can be induced by multiple signals and requires the CD44 cytoplasmic domain.

Human Jurkat T cells transfected with a human CD44H gene do not bind fluorescein-conjugated hyaluronic acid (F-HA). Activation of Jurkat CD44 transfectant is required for binding to F-HA. Binding can be induced by anti-CD3 monoclonal antibody (Mab), PMA, calcium ionophore, forskolin, or a monoclonal anti-CD44 Mab (F10-44-2). Cytochalasin D, an inhibitor of actin polymerization, inhibited both PMA-induced and anti-CD44 Mab-induced binding. In contrast, only PMA-induced binding was blocked by inhibitors of microtubule functions: colchicine or Taxol. Therefore, binding induced by PMA and anti-CD44 Mab involves different cytoskeletal proteins. The conclusion that interactions between CD44 and cytoskeletal proteins are important for binding was further supported by our observations that the cytoplasmic domain of CD44 is required for both PMA- and anti-CD44 Mab-induced binding. Jurkat CD44 mutant transfectant lacking the last 23 amino acids of the cytoplasmic domain can be induced to bind FHA. In contrast, Jurkat mutant CD44 transfectant lacking the last 57 amino acids of the cytoplasmic domain was unable to bind. Collectively, these data provide supportive evidence that binding activity of CD44 is under the regulation of multiple signal pathways and requires the presence of the cytoplasmic domain.

Amino Acid Sequence↗
PubMed1996

Genetic speciation of Candida isolates by arbitrarily primed polymerase chain reaction.

Candida species are opportunistic human pathogens capable of causing a variety of clinical diseases. Rapid and precise identification of Candida to species level is essential for effective treatment and management strategies. Conventional diagnosis of candidiasis is sometimes slow and variable. By using a random primer 5'-ACGGGCCAGT-3' in arbitrarily primed polymerase chain reaction, seven common Candida species (i.e., C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. lipolytica, C. tropicalis and C. (Torulopsis) glabrata) produced characteristic DNA band patterns, which enabled rapid determination to species level among them. Further analysis of the nucleotide sequences of Candida specific fragments should improve our understanding of the molecular structures and possible functions of the gene regions involved.

Candida↗
PubMed1996

Co-existence of serum-dependent and serum-independent mechanisms for liposome clearance and involvement of non-Kupffer cells in liposome uptake by mouse liver.

The effect of serum on liver uptake of liposomes with different compositions was investigated using a single-pass liver perfusion technique. Among the liposomes tested are those containing CL, PA, DPGS, PE or glycolipids such as PI, GD, GT1b and aGM1. Liposomes containing PA, CL and DPGS showed high level of liver uptake in the absence of serum. Presence of serum decreased the total liver uptake for liposomes containing CL and PA by 50% and did not affect the level of liver uptake for DPGS-containing liposomes. The presence of serum, however, significantly increased the liposome uptake by the perfused liver for PG, PE and aGM1 liposomes. Liposomes containing PI showed a minimal liver uptake regardless of serum presence. Fluorescence microscopic studies using a dual fluorescence label system in combination with Kupffer cell elimination technique showed that, in addition to the dominant role of Kupffer cells in taking up liposomes, non-Kupffer cells may also be involved in taking up CL and DCP-containing liposomes. Competition experiments using various liposome compositions indicated that liposome uptake by the liver cells may involve different receptors. Serum activity in enhancing the liver uptake for PE- and aGM1-containing liposomes can be blocked by treatment of serum with EDTA, EGTA/Mg2+ and high temperature (56 degrees C), suggesting the involvement of complement system. Results from this study support the conclusion that blood clearance of liposomes by the liver involves two independent mechanisms, one requires serum opsonins and the other does not.

Animals↗
PubMed1996

Effective doses of recombinant human bone morphogenetic protein-2 in experimental spinal fusion.

STUDY DESIGN: Nineteen dogs underwent L4-L5 intertransverse process fusions with either 58 micrograms, 115 micrograms, 230 micrograms, 460 micrograms, or 920 micrograms of recombinant human bone morphogenetic protein-2 carried by a polylactic acid polymer. A previous study (12 dogs) compared 2300 micrograms of recombinant human bone morphogenetic protein-2, autogenous iliac bone, and carrier alone in this model. All fusions subsequently were compared. OBJECTIVES: To characterize the dose-response relationship of recombinant human bone morphogenetic protein-2 in a spinal fusion model. SUMMARY OF BACKGROUND DATA: Recombinant osteoinductive morphogens, such as recombinant human bone morphogenetic protein-2, are effective in vertebrate diaphyseal defect and spinal fusion models. It is hypothesized that the quality of spinal fusion produced with recombinant human bone morphogenetic protein-2, above a threshold dose, does not change with increasing amounts of inductive protein. METHODS: After decortication of the posterior elements, the designated implants were placed along the intertransverse process space bilaterally. The fusion sites were evaluated after 3 months by computed tomography imaging, high-resolution radiography, manual testing, mechanical testing, and histologic analysis. RESULTS: As in the study using 2300 micrograms of recombinant human bone morphogenetic protein-2, implantation of 58-920 micrograms of recombinant human bone morphogenetic protein-2 successfully resulted in intertransverse process fusion in the dog by 3 months. This had not occurred in animals containing autograft or carrier alone. The cross-sectional area of the fusion mass and mechanical stiffness of the L4-L5 intersegment were not dose-dependent. Histologic findings varied but were not related to rhBMP-2 dose. Inflammatory reaction to the composite implant was proportional inversely to the volume of the fusion mass. CONCLUSIONS: No mechanical, radiographic, or histologic differences in the quality of intertransverse process fusion resulted from a 40-fold variation in dose of recombinant human bone morphogenetic protein-2.

Animals↗
PubMed1996

Two alternative promoters direct neuron-specific expression of the rat microtubule-associated protein 1B gene.

Microtubule-associated protein 1B (MAP1B) is a major constituent of the neuronal cytoskeleton that is expressed at high levels during early brain development and plays a role in axonal growth and neuronal plasticity. Previous studies suggested that the regulation of its gene expression is primarily at the transcriptional level. Thus, the characterization of the promoter region should help to define regulatory elements that control neuron-specific and developmental expression of the MAP1B gene. We have isolated genomic clones containing up to 11 kb of the upstream region of the rat MAP1B gene, sequenced approximately 1.8 kb upstream from the translation start codon, and identified several consensus sequences. These sequences include a consensus element common to several neuronal genes, a TCC repeat, a cAMP response element, and two TATA boxes that were 134 nucleotides apart from each other. S1 nuclease and RNase protection assays identified two corresponding groups of transcription initiation sites that were used selectively in distinct regions of the nervous system and during different stages of development. Transient transfection assays with neuronal and non-neuronal cell lines demonstrated that each TATA sequence and its corresponding adjacent region could independently direct neuron-specific expression of a reporter gene. Furthermore, the transcription of the reporter gene was initiated from the same sites as those of the MAP1B gene in vivo. These results suggest that two alternative and overlapping promoters, one inducible and the other constitutive, regulate the temporal and tissue-specific expression of the rat MAP1B gene.

Amino Acid Sequence↗
PubMed1996

Expression of hyaluronidase by tumor cells induces angiogenesis in vivo.

Hyaluronic acid is a proteoglycan present in the extracellular matrix and is important for the maintenance of tissue architecture. Depolymerization of hyaluronic acid may facilitate tumor invasion. In addition, oligosaccharides of hyaluronic acid have been reported to induce angiogenesis. We report here that a hyaluronidase similar to the one on human sperm is expressed by metastatic human melanoma, colon carcinoma, and glioblastoma cell lines and by tumor biopsies from patients with colorectal carcinomas, but not by tissues from normal colon. Moreover, angiogenesis is induced by hyaluronidase+ tumor cells but not hyaluronidase- tumor cells and can be blocked by an inhibitor of hyaluronidase. Tumor cells thus use hyaluronidase as one of the "molecular saboteurs" to depolymerize hyaluronic acid to facilitate invasion. As a consequence, breakdown products of hyaluronic acid can further promote tumor establishment by inducing angiogenesis. Hyaluronidase on tumor cells may provide a target for anti-neoplastic drugs.

Animals↗
PubMed1996

Isolation and sequencing of the 5' end of the rat microtubule-associated protein (MAP1B)-encoding cDNA.

We have isolated and sequenced the 5' end of the cDNA encoding the rat microtubule-associated protein 1B (MAP1B). We found that this region is highly homologous to the corresponding regions of the human [Lien et al., 22 (1994) 273-280] and mouse [Noble et al., J. Cell Biol. 109 (1989) 3367-3376] MAP1B genes. The combination of the sequence that we are presenting with the previously published sequence [Zauner et al., Eur. J. Cell Biol. 57 (1992) 66-74], represents the complete rat MAP1B cDNA coding sequence.

Animals↗
PubMed1996

Three-dimensional solution structure of the N-terminal domain of DNA polymerase beta and mapping of the ssDNA interaction interface.

DNA polymerase beta (beta-Pol) consists of an N-terminal ssDNA binding domain with deoxyribose phosphodiesterase activity and a C-terminal domain with nucleotidyltransferase activity. The solution structure of the cloned N-terminal domain of beta-Pol has been determined by multidimensional heteronuclear NMR using experimental restraints that included 1030 distances based on analysis of NOE connectivities, 68 phi, chi 1, and chi 2 torsion angles based on analysis of couplings, and 22 hydrogen bonds. Hydrogen bonds were assessed only within helices by the absence of saturation transfer from water at pH 6.7, by NOEs and JNH alpha couplings indicative of well-structured helices, and by 13C alpha chemical shifts characteristic of helices. The root mean square deviation for heavy backbone atoms within the helices was 0.64 A in 55 structures. The solution structure of the N-terminal domain is formed from four helices packed as two antiparallel pairs crossing at 50 degrees in a V-like shape. The domain binds p(dT)8, a template analogue, as a 1:1 complex in 100 mM NaCl (KD = 10 microM). Analysis of the binding equilibria at increasing NaCl concentrations indicated that ionic contacts contribute to the complex. The binding interaction was mapped to one face of the domain by characterizing backbone 1H and 15N chemical shift changes. Assigned intermolecular NOEs from 2D NOESY support the assessment of the binding interface. The structure that forms the interaction surface includes an antiparallel helix-3-turn-helix-4 motif and residues adjacent to an omega-type loop connecting helix-1 and helix-2. Sites appropriate for nucleotide contact on the structure are described. The mapped interaction interface for a ssDNA template is the first described for a DNA polymerase.

Amino Acid Sequence↗
PubMed1996

Distractive properties of a threaded interbody fusion device. An in vivo model.

STUDY DESIGN: Twenty sheep underwent anterior lumbar interbody fusions with either a threaded titanium interbody fusion device (cage, n = 8), autogenous iliac crest dowel graft (autograft, n = 6), or interbody decortication only (sham, n = 6). Two sheep had misplaced cages and were excluded. Sheep were killed after 6 months. OBJECTIVES: To determine whether this model is useful for examining the distractive and fixation properties of interbody fusion cages. SUMMARY OF BACKGROUND DATA: Interbody fusion cages are used in anterior lumbar interbody fusion procedures to provide immediate intersegmental fixation and to distract and preserve interbody height. The process of physiologic anchorage by bone ingrowth into such devices is under investigation. METHODS: Sheep were radiographed immediately after surgery and 2, 4, and 6 months after surgery. Interbody distraction and angulation were measured with a digital photo image analyzer at each time point. After the sheep were killed, stiffness to flexion, extension, and lateral bending moments were measured. Twelve untreated cadaver spines were also tested for comparison. RESULTS: After surgery, interbody distraction successfully occurred in cage and autograft-implanted sites. Loss of interbody height ensued, however, in all groups during the first 2 months. Percentage loss of height was lowest in cage sites. By 6 months, only cage sites remained distracted beyond normal. Fusions in all groups were stiffer than untreated spines. Autograft sites were stiffer than cage sites to lateral bending. Sham sites were stiffer than cage and autograft sites to flexion, but this likely resulted from complete intervertebral collapse. CONCLUSION: Despite early subsidence, interbody fusion cages successfully distracted and preserved interbody spaces. This model is useful for investigating methods of improving distraction and stabilization.

Animals↗
PubMed1996

A cysteine residue located in the transmembrane domain of CD44 is important in binding of CD44 to hyaluronic acid.

In the transmembrane domain and cytoplasmic domain of human CD44 protein there are two cysteine residues. These two cysteines are conserved in all known mammalian CD44 proteins. The functions of these cysteine residues are not known. Site-specific mutagenesis was used to create CD44 mutant proteins lacking either one or both of these cysteine residues. Wild-type CD44 and mutant CD44 genes were transfected into CD44- Jurkat cells to establish stable transfectants. These transfectants were used to study whether these two cysteine residues are important in the binding of CD44(H) to fluorescein-conjugated hyaluronic acid (F-HA). Jurkat transfectant bearing wild-type CD44 did not bind F-HA, unless they were stimulated in vitro with immobilized anti-CD3 monoclonal antibody. Anti-CD3 antibody also stimulated the binding of F-HA in Jurkat CD44.C295A transfectant in which the cytoplasmic cysteine residue has been replaced with alanine. In contrast, anti-CD3 antibody failed to stimulate the binding of F-HA in Jurkat transfectant (CD44.C286A), in which the transmembrane domain cysteine 286 has been replaced with an alanine, and in Jurkat transfectant CD44.2C2A, in which both of the cysteine residues have been altered. Binding can also be induced with a monoclonal anti-CD44 antibody (F-44-10-2) in Jurkat wild-type CD44 and Jurkat CD44.C295A transfectants but not in CD44. C286A transfectant. These results provide evidence that the transmembrane domain of CD44, more specifically the cysteine residue in the transmembrane domain, is important for both activation-induced and anti-CD44 antibody-induced binding of soluble HA.

Amino Acid Sequence↗
PubMed1996

Substrate specificity of glycinamide ribonucleotide synthetase from chicken liver.

Several analogs of glycinamide ribonucleotide and phosphoribosylamine have been prepared and evaluated as substrates for glycinamide ribonucleotide synthetase purified from chicken liver. Glycinamide ribonucleotide analogs include side chain modifications wherein the glycine side chain (R = CH2NH2) has been replaced by R = CH2NHCH3 and R = CH2CH2NH2, ribose ring replacement by chiral cyclopentane and cyclopentene derivatives, and phosphate replacement by phosphonates. All of these, with the exception of the O-phosphonate, served as substrates for the reverse enzymatic reaction, with Vmax values comparable to that obtained with glycinamide ribonucleotide, although the Km values ranged from 21 to 118 times the Km for glycinamide ribonucleotide. Analogs of phosphoribosylamine examined as substrates for the forward reaction consist of chiral derivatives of cyclopentane and cyclopentene and a chiral carbocyclic phosphonate. These also served as substrates, with Km values ranging from 5 to 23 times the Km for phosphoribosylamine and with diminished Vmax values. These studies have begun to define the structural features of the nucleotide substrate necessary to support enzymatic activity. Sarcosine (N-methylglycine) and beta-alanine were also accepted as substrates, albeit with reduced affinity compared with glycine.

Animals↗
PubMed1996

Localization of classical and alternative pathway regulatory activity within the decay-accelerating factor.

Decay-accelerating factor (DAF) is a cell-associated C regulatory protein that protects host cells from autologous C attack. It functions intrinsically in host cell surface membranes to rapidly dissociate autologous classical and alternative pathway C3 convertases whenever these amplifying enzymes assemble on host cell surfaces. It is composed of four contiguous approximately 70 amino acid long regions termed short consensus repeats (SCRs) that share homology with similar units in other C3 convertase regulatory proteins. It is attached to the cell surface membrane by a glycoinositol phospholipid (GPI) anchor that is added posttranslationally. In this study, we prepared rGPI-anchored DAF proteins devoid of individual SCRs. We then incorporated the GPI-anchored products into sheep erythrocyte (Esh) hemolytic intermediates and examined their abilities to intrinsically regulate classical or alternative pathway activation. We found that classical pathway C3 convertase regulatory function resides within SCR-2 and SCR-3, while alternative pathway C3 convertase regulatory function resides within SCR-2, -3, and -4. Functional comparisons of the variant DAF proteins in fluid phase C3 activation assays established that the differences reflect domain-specific interactions rather than changes in the spatial arrangement of SCRs above the cell surface. In accordance with these findings, we found that variant DAF molecules containing SCR-1, -2, and -3, but not SCR-4, function to selectively inhibit classical pathway activation.

Animals↗
PubMed1996

Substrate specificity of glycinamide ribonucleotide transformylase from chicken liver.

Several glycinamide ribonucleotide analogs have been prepared and evaluated as substrates and/or inhibitors of glycinamide ribonucleotide transformylase from chicken liver. The side chain modified analogs, in which the glycine side chain, R = CH2NH2, has been replaced by R = CH2NHCH3 and R = CH2CH2NH2, are substrates, with V/K (relative intensity) of 2.4% and 16.3%, respectively. Several carbocyclic analogs of glycinamide ribonucleotide, including the phosphonate derivative of carbocyclic glycinamide ribonucleotide, did not serve as substrates, but were inhibitors of the enzyme, competitive against glycinamide ribonucleotide, with Ki values ranging from 7.4 to 23.6 times the Km for glycinamide ribonucleotide. However, the O-phosphonate analog of carbocyclic glycinamide ribonucleotide did support enzymatic activity, with V/K (relative intensity) of 0.8%. In addition, glycinamide ribonucleoside was neither a substrate for, nor an inhibitor of, glycinamide ribonucleotide transformylase. Furthermore, alpha-glycinamide ribonucleotide had no effect on enzyme activity. These studies have begun to define the structural features of the nucleotide substrate required to support enzymatic activity.

Acyltransferases↗
PubMed1996

Use of arbitrarily primed polymerase chain reaction to differentiate Trichophyton dermatophytes.

Dermatophytes such as Trichophyton species are common human pathogens, the infection of which results in dermatophytosis (also known as ringworm). Several laboratory tests are used routinely for the diagnosis of dermatophytosis, but they are either slow or lacking specificity. Through examination of genomic DNA from Trichophyton dermatophytes and other fungi in arbitrarily primed PCR, it was shown that a random primer 5'-ACCCGACCTG-3' produced bands of 4.3 kb, 1.9 kb, 1.7 kb and 0.7 kb in T. rubrum DNA, bands of 2.5 kb, 1.9 kb and 0.8 kb in T. mentagrophytes var. interdigitale and T. mentagrophytes var. mentagrophytes DNA, and bands of 2.5 kb, 1.9 kb, 1.5 kb and 0.9 kb in T. tonsurans DNA. This primer amplified bands of different sizes in other fungal DNA. Therefore, based on the distinct band patterns observed in arbitrarily primed PCR using this primer, T. rubrum, T. mentagrophytes and T. tonsurans dermatophytes could be rapidly differentiated.

DNA Primers↗
PubMed1996

Serum independent liposome uptake by mouse liver.

The rate of liposome clearance from blood by the reticuloendothelial system (RES), primarily the Kupffer cells of the liver, depends largely on liposome composition. Inclusion of phosphatidylserine or dicetyl phosphate into liposomes with a simple composition of phosphatidylcholine and cholesterol increases liposome clearance, while inclusion of GM1 or amphipathic poly(ethylene glycol) decreases the rate of liposome clearance. To understand the underlying mechanism by which liposome clearance is regulated by the RES, we have developed a simple liver perfusion system. Using mouse liver as a model, we demonstrated that hepatic uptake of neutral or negatively charged liposomes does not involve serum components. Liver uptake of liposomes is directly related to the surface characteristics of liposomes. Liposomes with a neutral composition of phosphatidylcholine and cholesterol exhibit relatively low liver uptake. Inclusion of PS or DCP into these liposome dramatically enhances liposome uptake by the perfused liver. Conversely, inclusion of GM1 or PEG derivatives into liposomes greatly reduces the liposome uptake by the mouse liver. In contrast to the neutral or negatively charged liposomes, serum enhances the liver uptake of positively charged liposomes. Such serum effect on liver uptake of the positively charged liposomes is likely due to liposome aggregations caused by serum proteins. Inhibition of the liver uptake for PS-containing liposomes using liposomes with different compositions suggests that liver uptake of liposomes may involve different receptors.

Animals↗