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Biomedical subjects

D Lu

Publications and source records attributed to D Lu.

At least 271 records · Page 15Linked to original sources
PubMed1995

[Comparison of simple methods to screen predisposing eye of primary angle-closure glaucoma].

6,584 population over 40 years old in Canton universities and factories were screened for the predisposing eyes of primary angle-closure glaucoma (PACG). In the screening, three methods for the measurement of anterior chamber depth were used and compared and the critical limits of two simple ones of them [peripheral anterior chamber depth (PACD) and oblique illumination test with a flashlight] were evaluated. It was found that the incidence of PACG is as high as 10.0% in eyes with axial anterior chamber depth (AACD) < or = 2.0 mm, it is 6.7% in eyes with peripheral anterior chamber depth (PACD) < or = 1/4 corneal thickness (CT) and is 6.9% in eyes with iris light band ratio (ILBR) < or = 1/4. Also, we found that the incidence of PACG is as high as 77.8% in eyes with AACD < or = 1.6 mm and 100.0% in, AACD < or = 1.4 mm. The authors propose that PACD < or = 1/4 CT and ILBR < or = 1/4 can be the critical values of the two methods for the primary step of PACG screening.

Adult↗
PubMed1994

Molecular mechanism for familial protein C deficiency and thrombosis in protein CVermont (Glu20-->Ala and Val34-->Met).

The role of two protein C gamma-carboxyglutamic acid domain mutations in familial thrombosis, protein CVermont (Bovill, E. G., Tomczak, J. A., Grant, B., Bhushan, F., Pillemer, E., Rainville, I.R., and Long, G. L. (1992) Blood 79, 1456-1465), was investigated. Two single mutations (Glu20-->Ala and Val34-->Met) and the naturally occurring double mutation were created by site-directed mutagenesis and were expressed in human kidney 293 cells. Purified recombinant protein C with the mutation glutamate to alanine at position 20 is defective in the assays of activated partial thromboplastin time, factor Va inactivation, and fibrinolysis. Mutation from valine to methionine at position 34 has only a minor effect. Activation of Glu20 mutants by thrombin-thrombomodulin was not enhanced by phospholipid vesicles and showed a different calcium dependence compared with the wild type, suggesting that Gla20 is important in the interaction of the protein C Gla domain with a phospholipid-mediated site on the thrombomodulin molecule. Glu20-substituted protein C is not inhibited by calcium ion in its interaction with the calcium-dependent monoclonal antibody H-11, suggesting that this mutation has lost the calcium-induced, lipid-independent conformational transition of the protein C Gla domain. These data indicate that the loss of Gla20 causes the major familial dysfunction of protein C to associate with phospholipid as well as to undergo Ca(2+)-dependent, lipid-independent conformational changes and are consistent with the importance of Gla20 in both external and internal Ca2+ binding based upon the x-ray-derived structure of the homologous Gla domain in bovine prothrombin.

Alanine↗
PubMed1994

Agouti protein is an antagonist of the melanocyte-stimulating-hormone receptor.

The genetic loci agouti and extension control the relative amounts of eumelanin (brown-black) and phaeomelanin (yellow-red) pigments in mammals: extension encodes the receptor for melanocyte-stimulating hormone (MSH) and agouti encodes a novel 131-amino-acid protein containing a signal sequence. Agouti, which is produced in the hair follicle, acts on follicular melanocytes to inhibit alpha-MSH-induced eumelanin production, resulting in the subterminal band of phaeomelanin often visible in mammalian fur. Here we use partially purified agouti protein to demonstrate that agouti is a high-affinity antagonist of the MSH receptor and blocks alpha-MSH stimulation of adenylyl cyclase, the effector through which alpha-MSH induces eumelanin synthesis. Agouti was also found to be an antagonist of the melanocortin-4 receptor, a related MSH-binding receptor. Consequently, the obesity caused by ectopic expression of agouti in the lethal yellow (Ay) mouse may be due to the inhibition of melanocortin receptor(s) outside the hair follicle.

Adenylyl Cyclases↗
PubMed1994

Loss of membrane-dependent factor Va cleavage: a mechanistic interpretation of the pathology of protein CVermont.

Clinical manifestations of arterial and venous thrombosis in a family with protein C deficiency was associated with two mutations in the light chain of protein C: Glu20-->Ala and Val34-->Met. Further studies showed that the mutation Glu20-->Ala which eliminated a gamma-carboxylation site was exclusively responsible for the anticoagulant defect of activated protein C (APC). Membrane-bound human factor Va is inactivated by APC after two sequential cleavages of the heavy chain at Arg506 and Arg306. Human factor Va inactivation by human recombinant APC (rAPC) and a mutant molecule with an alanine instead of a glutamic acid at position 20 (rAPC(gamma 20A)) was investigated in the presence and absence of phospholipid vesicles. During a 2-hour incubation period of the cofactor with either rAPC or rAPC(gamma 20A). In the absence of a membrane surface, factor Va is cleaved quantitatively at Arg506 and retains approximately 60% of its initial cofactor activity. After a 2-hour incubation period with rAPC membrane-bound factor Va has no cofactor activity, whereas in the presence of a membrane surface and rAPC(gamma 20A) factor Va retains 60% of its initial cofactor activity. The completed loss in factor Va cofactor activity upon incubation of the membrane-bound cofactor with phospholipid vesicles and rAPC is associated with cleavages at Arg506 and Arg306, whereas membrane-bound factor Va cleavage at Arg306 by rAPC(gamma 20A) is impaired, resulting in a cofactor that is cleaved at Arg506. Slow cleavage at Arg306 occurs when membrane-bound factor Va is incubated with rAPC(gamma 20A) and only small amounts of fragments of M(r) = 45,000 and 30,000 are noticed. Our data show that the genetic defect which leads to the absence of a gamma-carboxylation site at Glu20 impairs membrane binding of human APC, which in turn is required for cleavage of factor Va at Arg306 and inactivation of the cofactor. The consequence of impaired membrane-dependent cleavage at Arg306 is manifested in vivo by venous and arterial thrombosis.

1-Carboxyglutamic Acid↗
PubMed1994

Structure of mouse protein S as determined by PCR amplification and DNA sequencing of cDNA.

The cDNA sequence of mouse protein S was derived by conventional PCR amplification from liver mRNA, initially using primers derived from the human cDNA sequence, followed by direct DNA sequencing. Seven overlapping PCR fragments covering all of the mature protein, part of the propeptide, and the 3' noncoding region were generated and sequenced. In some cases primers based upon the human cDNA sequence were ineffective. Subsequent successful amplification with mouse-derived primers to the same regions and comparison of the mouse and human sequences in these regions suggest that the failure of the human primers was due to insufficient degree of heterospecies identity. The mouse protein S cDNA sequence of the coding region shares 82% identity to human. The 3' noncoding region of mouse protein S cDNA has several small deletions and insertions compared to human protein S cDNA. Mature mouse protein S consists of 634 amino acids in a single polypeptide chain and displays domain organization similar to that for other species. The amino acid sequence of mouse protein S is about 80% identical to that of other species. Eleven glutamic acid residues were found in the amino terminal region and are predicted to be sites of gamma-carboxylation. Amino acid residues #80-244 are defined as four cysteine-rich repeat sequences homologous to epidermal growth factor. The remainder of the molecule is homologous to plasma sex steroid binding protein. The mouse protein S contains two potential N-glycosylation sites at positions #458 and 468 and is lacking the putative glycosylation site at #490 found in human protein S.

Amino Acid Sequence↗
PubMed1994

Receptor-mediated effects of angiotensin II on neurons.

Aside from its well-known and numerous actions at peripheral tissues, the octapeptide angiotensin II (ANG II) elicits specific receptor-mediated effects within the central nervous system. In this review we focus on the receptor-mediated actions of ANG II on neurons. The distribution of ANG II receptors in the brain and physiological, electrophysiological, and cellular effects mediated by these receptors are discussed. This is extended to a review of the characteristics of ANG II receptor subtypes on cultured neurons and the cellular and genomic actions mediated by these receptors. Finally, we develop this information into speculative models for the cellular effects mediated by each ANG II receptor subtype in neurons.

Angiotensin II↗
PubMed1994

Fabrication and characterization of glucose sensors based on a microarray H2O2 electrode.

A glucose sensor was recently developed by modifying an immobilized GOD (glucose oxidase) membrane and coating Nafion diffusion limiting membrane over an amperometric H2O2 MAE (microarray electrode), which was fabricated by using standard planar processing. The chip of the sensor is 2.5 mm by 15. The MAE is composed of an Ag/AgCl RE (reference electrode), a Pt CE (counter electrode) and two sets of Pt WE (working electrode), each set consists of five bands. Silicon nitride was chosen as the top insulator. Six MAEs different in both width and length of the band were examined and the MAE with the narrowest width (10 microns) of bands exhibits the best electrochemical performances. The glucose sensor based on the MAE B1-2 presents a linear range of 0.5-40 mM glucose at 37 degrees C with a 1/15 M phosphate buffer solution (pH = 7.0) as supporting electrolyte. The sensitivity, background current, response time, CV (coefficient of variation) and long-term stability of the sensor are 7.1 +/- 0.5 nA/mM, 2 +/- 0.5 nA, 30 s, 3.4% and 6 days respectively.

Biosensing Techniques↗
PubMed1994

Regulation of angiotensin II type 1 receptor mRNA in neuronal cultures of normotensive and spontaneously hypertensive rat brains by phorbol esters and forskolin.

Neuronal cells in primary culture from the brains of normotensive, Wistar-Kyoto (WKY) rats and spontaneously hypertensive (SH) rats express angiotensin II type 1 (AT1) receptors. Treatment of WKY rat brain cultures with a phorbol ester, phorbol 12-myristate 13-acetate (PMA), causes a time- and dose-dependent increase in the levels of an approximately 2.3-kb AT1 receptor mRNA transcript. A maximal stimulation of 4.5-fold in the AT1 receptor mRNA transcript level is observed with 200 nM PMA in 4 h and is blocked by 1 microM staurosporine. Forskolin also increases the AT1 receptor mRNA levels in WKY rat brain neurons in a time- and dose-dependent manner, and a 4.5-fold stimulation is achieved with 50 microM forskolin in 4 h. The stimulatory effects of both PMA and forskolin are completely abolished by coincubation of neuronal cultures with 1 microM actinomycin D. In addition, nuclear run-on assay indicated an increase in the transcription of AT1 receptor mRNA in WKY rat brain neurons treated with either PMA or forskolin. Both PMA and forskolin also stimulate levels of AT1 receptor mRNA in neuronal cultures from brain of the SH rat. The degree of stimulation in these cultures is comparable to that in WKY rat brain neurons. These observations show that although the basal AT1 receptor gene expression is significantly higher in SH rat brain neurons compared with WKY rat brain neurons, the protein kinase C- and protein kinase A-responsive stimulation is not altered. These data suggest a possible involvement of protein kinase C and protein kinase A response elements in AT1 receptor gene expression.

Animals↗
PubMed1994

Airway provocation test with ragweed pollen extract in Chinese asthmatics.

As ragweed becomes widespread over China, ragweed pollenosis tends to be more frequent. Incidence of ragweed pollenosis is about 1.04% in Qingdao district. To elucidate ragweed as an allergen in the development of bronchial asthma, ragweed pollen extract inhalation airway provocation tests (APT) were studied in 55 patients with ragweed pollenosis. A single dosage of 9,000 PNU ragweed pollen extract was applied and positive response was defined as more than 35% fall in SGrs at the dose of PT35-Grs. Among 55 patients, positive APTs were found in 27 cases (49%), dual late-phase responses in 11 patients, single late-phase responses in 2 patients in the complicated asthma group. Late-phase responses were related to the increased specific IgE and strong positive skin test response (wheal > 11 mm). The results suggested ragweed pollen was one of the important allergens causing bronchial asthma. There was airway hyperresponsiveness to ragweed pollen in some patients (20%) with ragweed pollenosis without asthmatic symptoms.

Adolescent↗
PubMed1994

Optimization of methods to achieve mRNA-mediated transfection of tumor cells in vitro and in vivo employing cationic liposome vectors.

Direct in vivo transfection of tumor nodules in situ via liposome-DNA complexes has been employed as a strategy to accomplish antitumor immunization. To circumvent the potential safety hazards associated with systemic localization of delivered DNA, the utility of mRNA transcript-mediated gene delivery was explored. Capped, polyadenylated mRNA transcripts encoding the firefly luciferase and Escherichia coli lacZ reporter genes were derived by in vitro transcription. Transfection of the human breast cancer cell line MDA-MB-435 in vitro was accomplished employing cationic liposome-mRNA complexes. Evaluation of a panel of cationic liposome preparations demonstrated significant differences in the capacity of the various preparations to accomplish mRNA-mediated transfection. Quantitative evaluation of in vitro transfection demonstrated that target cells could be transfected at a high level of efficiency. The mRNA liposome-complexes were evaluated for in vivo transfection of tumor nodules in human xenografts in athymic nude mice. It could be demonstrated the liposome-mRNA complexes were comparable in efficacy to liposome-DNA complexes in accomplishing in situ tumor transfection. Thus, mRNA may be considered as an alternative to plasmid DNA as a gene transfer vector for genetic immunopotentiation applications.

Breast Neoplasms↗
PubMed1994

Ribozyme-mediated in vitro cleavage of transcripts arising from the major transforming genes of human papillomavirus type 16.

Human papillomaviruses (HPV) have been strongly implicated as important cofactors in the development of several human malignancies, particularly anogenital carcinomas. Products arising from the E6 and E7 open reading frames (ORFs) from HPV-16, a type commonly associated with human cervical carcinoma, are essential for viral transformation. Unfortunately, a highly effective treatment for this infection is not available. To develop a novel treatment for this disease, ribozymes were designed to cleave all transcripts encoding HPV-16 E6 and E7 ORFs in proximity to their translational start sites ("AUG"). Cleavage sites for Rz110 and Rz558 occur immediately 3' to nucleotides 110 and 558 of the viral genomic DNA, respectively. Oligonucleotides corresponding to these ribozymes were synthesized and inserted into a eucaryotic viral vector derived from the nonpathogenic parvovirus, adeno-associated virus. Ribozyme transcription from this vector, termed CWRT7:SVN, is under control of both the highly active Rous sarcoma virus long terminal repeat and bacteriophage T7 promoters. T7 transcripts of the E6 and E7 ribozymes efficiently cleaved their cognate targets in vitro under a variety of conditions, including physiological temperature. These results may provide the basis for the development of a ribozyme-based, gene therapeutic treatment for HPV-associated diseases.

Avian Sarcoma Viruses↗
PubMed1994

Expression and immunogenicity of tripeptide repeat region on P190 of Plasmodium falciparum.

A DNA fragment, designated as P190TR, encoding amino acid residues of the tripeptide region of the P190 antigen was amplified by polymerase chain reaction from genomic DNA of FCC1/HN Plasmodium falciparum isolated from Hainan Province, China. Upon comparison with the nucleotide sequences of MAD20 allele, it was found that there were five bases substitution in the P190TR which cause amino acid changes. The DNA fragment sequenced were ligated to BamHI and XbaI-digested pGEX-2T vector. Competent E. coli JM109 (DE3) were transformed with either parental or recombinant pGEX-2T for expression. Analysis of soluble cellular proteins revealed the high level expression of GST-P190TR as fusion proteins. Affinity purification of the fusion protein under nondenaturing condition resulted in the removal of almost all other E. coli proteins. The purified P190TR protein was highly immunogenic in rabbits. The antibodies against the recombinant protein recognized the malaria parasite with the titers at 1:320 measured by IFA and antisera from malarial patients reacted with the expressed protein in Western Blot.

Amino Acid Sequence↗
PubMed1993

Palladium-103 versus iodine-125 for ophthalmic plaque radiotherapy.

PURPOSE: A dosimetry study comparing the use of I-125 vs. Pd-103 radioactive seeds for ophthalmic plaque brachytherapy. METHODS AND MATERIALS: Palladium-103 (Pd-103) seeds in ophthalmic plaques were used to treat 15 patients with intraocular malignant melanoma. Computer-aided simulations were performed to evaluate the intraocular dose distribution of I-125 versus Pd-103 ophthalmic plaques (delivering equivalent apex doses). Seven target points were selected. Starting at the outer scleral surface, four were located along the central axis of the plaque: the 1 mm point (the inner sclera), the 6 mm point, the tumors apex, and the opposite eye wall. We also evaluated the fovea, optic nerve, and the lens because they were considered to be critical structures. RESULTS: These studies demonstrated that the lower energy photons generated by Pd-103 seeds (average 21 KeV) in ophthalmic plaques were more rapidly absorbed in tissue than photons generated by I-125 (average 28 KeV). Therefore, during ophthalmic plaque radiotherapy, Pd-103 photons were found to be more rapidly absorbed within the tumor and less likely to reach most normal ocular structures. On average, the use of Pd-103 decreased the dose to the fovea by 5.7%, to the optic nerve by 8.4%, to the lens by 26%, and to the opposite eye wall by 38.4%. CONCLUSION: Palladium-103 ophthalmic plaque brachytherapy resulted in slightly more irradiation of the tumor and less radiation to most normal ocular structures.

Brachytherapy↗
PubMed1993

Cell lines from grc congenic strains of rats having different susceptibilities to chemical carcinogens.

The growth and reproduction complex (grc-) strains of rats have a 70-kilobase deletion in the major histocompatibility complex (MHC)-linked grc-G/C region that is associated with embryonic death, developmental defects, and an increased susceptibility to chemical carcinogens. To study further the effects associated with the deletion, fibroblastic cell lines from grc-, grc+, and grc+/- rat embryos were developed: BIL-derived cell lines are congenic for the MHC and grc, whereas R16-derived cell lines are congenic for the grc alone. In early passages, all cell lines expressed the MHC class I antigen RT1.A, had a diploid chromosome number, and did not display anchorage-independent growth or in vivo tumorigenicity. The grc- cells [median population doubling time (PDT), 47 h] grew more slowly than the grc+ (PDT, 30.5 h) and grc+/- (PDT, 33 h) cells. All cells underwent crisis, but the crisis stage began earlier and lasted longer in the grc- cells. The established grc- cell lines (PDT, 32.5 h) grew faster than the grc+ (PDT, 48.5 h) and grc+/- (PDT, 54 h) cell lines. Two of the three BIL-derived grc- lines that survived crisis became anchorage independent in tissue culture and tumorigenic in histocompatible F1 rats (highly malignant fibrosarcomas) at passages 33 and 48, respectively; by contrast, none of the R16-derived grc- cell lines transformed. None of 8 grc+ or 8 grc+/- cell lines that survived crisis displayed anchorage-independent growth or tumorigenicity under the same conditions up to passage 50. All of the established cell lines, including the two tumorigenic ones, expressed MHC class I antigens. Southern and Northern blot analyses of BIL-derived cell lines before and after crisis showed that they all constitutively expressed H-ras and Rb and that no cell line showed rearrangement, amplification, or overexpression of c-myc, H-ras, Rb, and p53 either before or after crisis. These observations indicate that: (a) the homozygous grc- deletion is necessary but not sufficient for in vitro transformation; (b) another genetic factor(s) required for transformation is linked to, or possibly in, the MHC; and (c) passage through crisis, spontaneous transformation, or carcinogen treatment does not alter the cellular expression of MHC class I antigens or of several oncogenes and tumor suppressor genes.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1993

Tyrosine phosphorylation of P160 BCR by P210 BCR-ABL.

It is well established that the chimeric BCR-ABL gene formed by joining parts of the BCR and ABL genes plays a key role in the pathogenesis of Philadelphia (Ph) chromosome-positive leukemias. We report that simultaneous expression of P210 BCR-ABL and P160 BCR in simian COS-1 cells yielded stable complexes of these two proteins, and induced phosphorylation of P160 BCR on tyrosine residues in vivo. Tyrosine phosphorylation of a deletion mutant encoding 553 amino acids of BCR N-terminal sequences was also detected when it was coexpressed with P210 BCR-ABL. We propose that tyrosine phosphorylation of P160 BCR by P210 BCR-ABL and their stable physical interaction may perturb normal BCR functions and that these alterations are directly involved in the pathologic processes found in Ph chromosome-associated leukemias.

Amino Acid Sequence↗
PubMed1993

Induction of differential heat shock gene expression in heart, lung, liver, brain and kidney by a sympathomimetic drug, amphetamine.

Amphetamine, a sympathomimetic drug, can elevate the body temperature by causing enhancement of endogenous lipolysis. This study was designed to examine whether this drug could induce the expression of the genes of heat shock proteins (HSP) in different tissues. Subcutaneous administration of amphetamine (10 mg/kg wt) to the rats increased the rectal temperature of the rats to 42 degrees C within 60 min. After 3 hr rats were sacrificed; and heart, lung, liver, kidney and brain tissue were removed to examine the induction of mRNAs for HSPs by Northern blot analysis using cDNA probes of 27 kDa, 70 kDa and 89 kDa HSPs. The results of this study indicate striking regional and cell type differences in the pattern of induction of the HSP mRNAs by amphetamine suggesting that different organs and cell types respond differently to amphetamine.

Amphetamine↗
PubMed1993

Effect of glycosphingolipid fatty acid chain length on behavior in unsaturated phosphatidylcholine bilayers: a 2H NMR study.

Deuterium-labeled glycosphingolipids, N-lignoceroyl[d47]galactosylceramide (24:0 fatty acid form) and N-stearoyl[d35]galactosylceramide (18:0 fatty acid form) were prepared by partial synthesis. These probe-labeled species, differing only in the lengths of their fatty acids, were compared via 2H NMR with regard to arrangement and behavior in bilayers of the monounsaturated phospholipid 1-stearoyl-2-oleoylphosphatidylcholine (SOPC). Results were used to consider the physical significance of the great range of common acyl chain lengths that is a frequent feature of cell membrane glycosphingolipids. N-lignoceroyl[d47]- and N-stearoyl[d35]galactosylceramide were incorporated at concentrations ranging from 5 to 50 mol % into unsonicated phospholipid liposomes, and their spectra were analyzed in the range +73 to -14 degrees C. For the 18:0 fatty acid derivative, first spectral moments, M1, were calculated and plotted as a function of temperature for each sample composition. Spectral inspection for regions of phase coexistence, in conjunction with consideration of M1 curves, permitted derivation of phase diagram boundaries which were then refined using spectral subtraction techniques. The phase diagram for galactosylceramide with short fatty acid in SOPC was compared to the corresponding phase diagram for its long-chain analogue, derived previously in the same fashion [Morrow, M. R., Singh, D., Lu, D., & Grant, C. W. M. (1992) Biochim. Biophys. Acta 1106, 85-93]. The binary phase diagrams referred to above, which reflect the behavior of short- and long-chain glycolipids in a common phospholipid host matrix, displayed important similarities and differences. In fluid membranes, the behavior appeared to be remarkably alike, as reflected in superimposable fluidus curves over the concentration range studied.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗