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Biomedical subjects

D Liu

Publications and source records attributed to D Liu.

At least 343 records · Page 19Linked to original sources

Nucleotide sequence of a 5892 base pairs fragment of the LsMNPV genome and phylogenetic analysis of LsMNPV.

A 5892 bp fragment of Leucania separata multiple nuclear polyhederosis virus (LsMNPV) containing gp37, 39 k genes and another four ORFs was sequenced in this article. According to regulatory elements on 5' uncoding sequences, the ORF4 and ORF1 are probably two novel baculovirus genes, and the ORF4 probably also is a new early-late gene. The possible functions about GP37 and 39 K proteins were explored. The homology of GP37 protein among LsMNPV and 8 other insect virus was compared. The evolutional position of LsMNPV is also estimated based on the homology of gp37 genes. The structure and its homology of several genes in LsMNPV prove that LsMNPV is far related with other baculovirus and has an exceptional genome organization.

Amino Acid Sequence↗

Solution structure of the homodimeric core domain of Escherichia coli histidine kinase EnvZ.

Escherichia coli osmosensor EnvZ is a protein histidine kinase that plays a central role in osmoregulation, a cellular adaptation process involving the His-Asp phosphorelay signal transduction system. Dimerization of the transmembrane protein is essential for its autophosphorylation and phosphorelay signal transduction functions. Here we present the NMR-derived structure of the homodimeric core domain (residues 223-289) of EnvZ that includes His 243, the site of autophosphorylation and phosphate transfer reactions. The structure comprises a four-helix bundle formed by two identical helix-turn-helix subunits, revealing the molecular assembly of two active sites within the dimeric kinase.

Amino Acid Sequence↗

Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA.

Development of methods that allow an efficient expression of exogenous genes in animals would provide tools for gene function studies, treatment of diseases and for obtaining gene products. Therefore, we have developed a hydrodynamics-based procedure for expressing transgenes in mice by systemic administration of plasmid DNA. Using cDNA of luciferase and beta-galactosidase as a reporter gene, we demonstrated that an efficient gene transfer and expression can be achieved by a rapid injection of a large volume of DNA solution into animals via the tail vein. Among the organs expressing the transgene, the liver showed the highest level of gene expression. As high as 45 microg of luciferase protein per gram of liver can be achi- eved by a single tail vein injection of 5 microg of plasmid DNA into a mouse. Histochemical analysis using beta-galactosidase gene as a reporter reveals that approximately 40percent of hepatocytes express the transgene. The time-response curve shows that the level of transgene expression in the liver reaches the peak level in approximately 8 h after injection and decreases thereafter. The peak level of gene expression can be regained by repeated injection of plasmid DNA. These results suggest that a simple, convenient and efficient method has been developed and which can be used as an effective means for studying gene function, gene regulation and molecular pathophysiology through gene transfer, as well as for expressing proteins in animals.

Animals↗

D4 dopamine receptor-mediated phospholipid methylation and its implications for mental illnesses such as schizophrenia.

Previous studies have shown D2-like dopamine receptor involvement in the regulation of phospholipid methylation (PLM), while others have documented impaired methionine and folate metabolism in schizophrenia. Utilizing [14C]formate labeling in cultured neuroblastoma cell lines, we now show that D4 dopamine receptors (D4R) mediate the stimulatory effect of dopamine (DA) on PLM. The effect of DA was potently blocked by highly D4R-selective antagonists and stimulated by the D4R-selective agonist CP-226269. DA-stimulated PLM was dependent upon the activity of methionine cycle enzymes, but DA failed to increase PLM in [3H]methionine labeling studies, indicating that a methionine residue in the D4R might be involved in mediating PLM. A direct role for MET313, located on transmembrane helix No. 6 immediately adjacent to phospholipid headgroups, was further suggested from adenosylation, site-directed mutagenesis and GTP-binding results. A comparison of PLM in lymphocytes from schizophrenia patients vs control samples showed a four-fold lower activity in the schizophrenia group. These findings reveal a novel mechanism by which the D4R can regulate membrane composition. Abnormalities in D4R-mediated PLM may be important in psychiatric illnesses such as schizophrenia.

Amino Acid Sequence↗

Detection of bone marrow micrometastasis.

The detection of metastasizing single tumor cells has so far been difficult. Using a monoclonal antibody (MAb A45-B/B3) recognizing human cytokeratin, we identified immunocytochemically single tumor cells and micrometastases in patients (n = 24) with nonsmall cell lung cancer at the time of surgery of the primary tumor. The cytokeratin-positive cells (1-14/5 x 10(5) cells) in the bone marrow samples of 9 (9/24) patients were found. We also found a garland-like cluster, which consists of seven cancer cells and two closely connected tumor cells from one bone marrow sample. These results indicate that this technique can be used as a early diagnostic technique of bone marrow micrometastasis in the patient with the nonsmall cell lung cancer.

Animals↗

The roles of free radicals in amyotrophic lateral sclerosis: reactive oxygen species and elevated oxidation of protein, DNA, and membrane phospholipids.

To explore whether reactive oxygen species (ROS) play a role in the pathogenesis of amyotrophic lateral sclerosis (ALS), a unique microdialysis or microcannula sampling technique was used in mice transfected with a mutant Cu,Zn-superoxide dismutase (SOD1) gene from humans with familial ALS, mice transfected with the normal human SOD1 gene, and normal mice. We demonstrate for the first time that the levels of hydrogen peroxide (H(2)O(2)) and the hydroxyl radical ((.)OH) are significantly higher, and the level of the superoxide anion (O(2)(.-)) is significantly lower in ALS mutant mice than in controls, supporting by in vivo evidence the hypothesis that the mutant enzyme catalyzes (.)OH formation by the sequence: O(2)(.-) --> H(2)O(2) --> (.)OH. This removes doubts regarding the relevance of elevated ROS in FALS raised by in vitro experiments. The levels of oxidation products are also significantly higher in the mutant mice than in controls, consistent with some previous reports. Only the superoxide concentration differs between two controls among all the measurements. Our findings correlate in vivo a gene mutation to both elevated H(2)O(2) and (.)OH and increased oxidation of cellular constituents. The elevated H(2)O(2) in mutant mice indicates impairment of its detoxification pathways, perhaps by changed interactions between SOD1 and H(2)O(2) detoxification enzymes.-Liu, D., Wen, J., Liu, J., Li, L. The roles of free radicals in amyotrophic lateral sclerosis: reactive oxygen species and elevated oxidation of protein, DNA, and membrane phospholipids.

Amyotrophic Lateral Sclerosis↗

Fatal injuries in the United States construction industry involving cranes 1984-1994.

There is little published information concerning the epidemiology of injuries in the construction industry involving cranes other than for electrical injury from power line contact. For the 11-year period of 1984 through 1994, the US Occupational Safety and Health Administration (OSHA) investigated 502 deaths in 479 incidents involving cranes in the construction industry. Electrocution was the largest category, with 198 deaths (39%) reported. Other major categories were assembly/dismantling (58 deaths, 12%), boom buckling (41 deaths, 8%), crane upset/overturn (37 deaths, 7%), and rigging failure (36 deaths, 7%). The majority of the deaths during assembly/dismantling involved removal of the boom pins from lattice boom cranes. Only 34% of the construction firms employing the fatally injured workers had ever been inspected by OSHA. OSHA cited the employer for safety violations in 436 deaths (83%). Additional worker training, increased OSHA inspections, and crane inspection programs could prevent many crane-related deaths.

Accidents, Occupational↗

The integrin alpha10 subunit: expression pattern, partial gene structure, and chromosomal localization.

Herein we report the cloning of cDNAs and incompletely processed hnRNAs from endothelia and heart that encode the alpha10 subunit forming part of the novel collagen type II-binding integrin alpha10beta1 of chondrocytes. Analysis of hnRNA clones and reported expressed sequence tags revealed the positions of 17 putative intron-exon splice junctions shared with those of the p150,95 (ITGAX) gene. Human alpha10 transcripts of 5.4 and 1.8 kb were not restricted to chondrocytes but, instead, were widely expressed in a panel of 24 tissue types, where the highest expression was found in muscle and heart. The human alpha10 subunit gene (ITGA10) was localized to band q21 of chromosome 1.

Amino Acid Sequence↗

Generation of stable recombinant retroviruses containing the beta-globin genes linked to complex regulatory elements by using transient transfection.

A transient packaging method for a retroviral vector containing beta-globin sequences, mediated by Ca3(PO4)2 precipitation, generated stable recombinant beta-globin retroviruses with intact viral genomes. We suggest the use of this method as an alternative means to overcome the production of beta-globin encoding retroviruses with aberrant viral genomes using the routine packaging procedure.

Genetic Vectors↗

Effects of cyclosporin A on alveolar bone: an experimental study in the rat.

BACKGROUND: There have been several investigations on the role of cyclosporin A (CSA) in gingival hyperplasia in both animals and humans. However, less attention has been given to the drug's effect on alveolar bone. This study used light microscopy to histologically and histometrically evaluate the effects of CSA on alveolar bone in the rat. METHODS: Sixty, 6-week-old Sprague-Dawley rats were separated into test and control groups. Animals in the test group received CSA in mineral oil (30 mg/kg body weight) daily by gastric feeding over the 6-week study. Control animals received only mineral oil. Ten animals from each group were sacrificed at weeks 2, 4, and 6. After histologic processing, the labial crest of the alveolar bone in the anterior mandible was evaluated by light microscopy. RESULTS: A distinct pattern of increased osteoclasia and reduced bone formation was observed in the CSA-exposed animals compared to the controls. Increased osteoclasia was observed in periodontal sites and decreased bone formation was observed in symphyseal sites. CONCLUSIONS: The results suggest that CSA has distinct effects on alveolar bone.

Alveolar Bone Loss↗

Synthesis and characterization of aromatic ring-based cationic lipids for gene delivery in vitro and in vivo.

A new series of cationic lipids has been synthesized for gene delivery using 3,5-dihydroxybenzyl alcohol as the backbone and starting material. Using CMV driven expression system and luciferase gene as a reporter, we demonstrated that the transfection activity of these new lipids when formulated with Tween 80 as co-lipid is comparable to that of DOTAP, one of the most commonly used cationic lipids for transfection. Among the four different cell lines tested including murine melanoma BL-6 cells, human embryonic kidney 293 cells, HepG2 and HeLa cells, the highest transgene expression was seen in 293 cells. Results from in vivo experiments using mice as an animal model show that these cationic lipids preferentially transfect the cells in the lung upon tail vein administration. The cationic lipid, N,N,N-trimethyl-N-[3,5-bis(tetradecyloxy)benzyl] ammonium bromide 4c(di-C14:0) with two 14-hydrocarbon chains exhibits the best transfection activity. These results suggest that these new aromatic ring-based cationic lipids are useful transfection reagents for both in vitro and in vivo gene transfer studies.

Animals↗

Evidence-based surgery and the internet.

The preliminary version of the Evidence-Based Surgery (EBS) web pages was first released by the College Library in December 1998. In just four weeks there were over 1,600 recorded 'hits'. These draft pages were made known only to those surgeons and trainees participating on the various surgical e-mail discussion forums such as the Association of Surgeons in Training Mailing Lists. The popularity of the draft EBS pages and the encouraging feedback has rendered the on-going updating of the EBS pages a worthwhile activity.

Computer Communication Networks↗

The role of plasminogen activator receptor in cancer invasion and dormancy.

Urokinase plasminogen activator receptor (uPAR) has been identified some 15 years ago and the anticipation was that its presence on the cell surface will provide a focus for anchoring uPA and possibly protect the enzyme from native inhibitors. The studies of the last decade have shown that uPA localized to the surface of cells by uPAR is indeed an important factor in the process of cancer cell invasion and metastasis. We developed a chick embryo model in which we showed that uPAR is crucial in invasion of stroma and in intravasation (breaching of the blood vessels walls). More recently and unexpectedly, uPAR--a protein anchored in the outer leaf-let of the plasma membrane, has been shown to initiate signal transduction events and affect cell migration. We have shown that uPAR co-associates with fibronectin binding integrin, alpha 5 beta 1, activates them and that this interaction leads to a greatly increased level of active ERK. When the association between uPAR and integrin or integrin and fibronectin are interrupted either by reduction of surface uPAR expression, or by other means, human carcinoma cells enter a state of protracted dormancy. We show that very high levels of active ERK are required to keep cancer cells proliferating in vivo.

Animals↗