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Biomedical subjects

D Sun

Publications and source records attributed to D Sun.

At least 181 records · Page 10Linked to original sources
PubMed1999

[Study on the inclusion effect of menthol with beta-cyclodextrin].

Indusion effect of beta-CD with menthol was studied by three methods of preparation. Determination of physical constants and quantitative analysis proved that the inclusion compounds of beta-CD with menthol were formed, which was found to be 1:1 and 2:3. The results of thermal analysis demmonstrated that the gasifiation temperature of the inclusion compounds was by far higher than that of the free menthol. It was considered that the inclusion compound must be disconnected to be free, and then it can be determined by gas chromatography in the same condition of the free menthol.

Chromatography, Gas↗
PubMed1999

[Effects of anion ions on the structures of lanthanum glycine amino acid complex compounds].

Two lanthanum-glycine amino acid complexes, La(Gly)3 (ClO4)3 x H2O and La(Gly)3Cl3 x 3H2O, were synthesized with different anion ions. The infrared spectra of two complexes were characterized. The results show that the two complexes have different structures with different anion ions and the water molecules exit with different status in the two complexes. The results also indicate that one of the perchlorate ions is involved in the coordination in La(Gly)a(ClO4)3 x H2O.

Anions↗
PubMed1998

Human 12(R)-lipoxygenase and the mouse ortholog. Molecular cloning, expression, and gene chromosomal assignment.

Expressed sequence tag information was used to clone the full-length sequence for a new human lipoxygenase from the B cell line CCL-156. A related mouse sequence with 83% nucleotide identity to the human sequence was also cloned. The human lipoxygenase, when expressed via the baculovirus/insect cell system produced an approximately 80-kDa protein capable of metabolizing arachidonic acid to a product identified as 12-hydroxyeicosatetraenoic acid by mass spectrometry. Using chiral phase-high performance liquid chromatography, the product was identified as >98% 12(R)-hydroxyeicosatetraenoic acid as opposed to the S-stereoisomer formed by all other known mammalian lipoxygenases. The single copy human 12(R)-lipoxygenase gene was localized to the chromosome 17p13 region, the locus where most other lipoxygenase genes are known to reside. By reverse transcription-polymerase chain reaction, but not by Northern blot, analysis the 12(R)-lipoxygenase mRNA was detected in B cells and adult skin. However, the related mouse lipoxygenase mRNA was highly expressed in epidermis of newborn mice and to a lesser extent in adult brain cortex. By in situ hybridization the mouse lipoxygenase gene was demonstrated to be temporally and spatially regulated during embryogenesis. Expression was induced at embryonic day 15.5 in epidermis, nasal epithelium, and surface of the tongue. These results broaden the mammalian lipoxygenase family to include a 12(R)-lipoxygenase whose biological function remains to be determined.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1998

[Effects of Trp64Arg mutation in the beta 3-adrenergic receptor gene on body fat, plasma glucose level, lipid profile, insulin secretion and action in Chinese].

OBJECTIVE: To ascertain the association of the Trp64Arg mutation in the beta 3-adrenergic receptor (ADRB3) gene with the total body fat and fat distribution, plasma glucose level, lipid profile, insulin secretion and action in Chinese. METHODS: Genotyping of the Trp64Arg mutation in the ADRB3 gene as well as measurment of clinical variables body mass index(BMI), percent of desirable weight(PDW), area of regional adipose tissue depots, fasting lipid profile, and also, plasma glucose, insulin, C-peptide, free fatty acids levels before and after glucose challenge and blood pressure were performed in 308 Chinese, of whom, 161 were nondiabetics and 147, type 2 diabetic subjects. RESULTS: (1) The frequency of Arg64 allele of ADRB3 gene in Chinese was similar to that in Japanese but slightly higher than that in Caucasians. (2) No association of any of the clinical varaibles with Trp64Arg mutation was observed in nondiabetics. (3) The Trp64Arg mutation in the ADRB3 gene was associated with BMI(P=0.019), PDW(P=0.001) and waist circumference (P=0.045) in type 2 diabetic subjects. In female diabetic subjects, more significant association with BMI(P=0.001), PDW(P=0.001), waist circumference(P=0.023), and also, association with abdominal and femoral subcutaneous fat area(P=0.032 and P=0.023) were observed. The frequencies of Arg64 allele were 21%, 14% and 4% in female diabetics with obesity, over-weight and normal-weight respectively (P=0.026). Logistic regression analysis showed that BMI was the only independent clinical variable related to the Trp64Arg mutation in the ADRB3 gene (P=0.0027). CONCLUSION: The Trp64Arg mutation in ADRB3 gene contributes to the variation of total body fat in Chinese type 2 diabetic subjects.

Adipose Tissue↗
PubMed1998

Superoxide released to high intra-arteriolar pressure reduces nitric oxide-mediated shear stress- and agonist-induced dilations.

It is thought that elevated levels of reactive oxygen metabolites contribute to the dysfunction of vascular endothelium in hypertension. We hypothesized that high intravascular pressure itself elicits production of superoxide, which then interferes with nitric oxide (NO)-mediated responses of arterioles. Thus, isolated arterioles (approximately 80 microm in diameter) from gracilis muscle of normotensive Wistar rats were cannulated and exposed to 140 mm Hg perfusion pressure for 30 minutes (in the absence of perfusate flow). After high intravascular pressure treatment, dilations to increases in perfusate flow (0 to 30 microL/min) were significantly reduced (from 39+/-2.2 to 19+/-2.1 microm at 30 microL/min), eliciting an increase in wall shear stress from approximately 20 to approximately 60 dyne/cm2. Nomega-nitro-L-arginine (10(-4) mol/L) did not affect, whereas indomethacin eliminated, flow-induced dilations after pressure treatment. In control, substance P (SP, 10(-9) to 5x10(-8) mol/L), sodium nitroprusside (SNP, 10(-8) to 10(-6) mol/L), and adenosine (ADO, 10(-6) to 5x10(-5) mol/L) elicited dilations (SP: 31.5+/-1.9 microm, SNP: 45.6+/-4 microm, and ADO: 37.2+/-4.1 microm, at maximum concentrations, respectively). After pressure treatment, maximum dilations to SP and SNP were significantly reduced (by 49% and 39%, respectively), whereas responses to ADO were not affected. Presence of superoxide dismutase (120 U/mL) and catalase (80 U/mL), but not catalase alone, in the perfusate solution prevented the reduction in dilation of arterioles to flow and agonists after pressure treatment by restoring NO mediation. We conclude that high intravascular pressure per se elicits the release of superoxide, which then interferes with NO, a mechanism that contributes to the elevation of wall shear stress and peripheral resistance in hypertension.

Adenosine↗
PubMed1998

Peptide sequence of an antibiotic cecropin from the vector mosquito, Aedes albopictus.

We have identified a 35-amino acid antibiotic cecropin secreted by an established mosquito cell line. C7-10 cells from the vector mosquito, Aedes albopictus, were incubated with heat-killed Escherichia coli, and materials secreted into the cell culture supernatant were recovered by acid precipitation. Following batch elution from Sep-Pak C18 cartridges and further purification by reverse phase high performance liquid chromatography (RP-HPLC) a predominant peak of antibacterial activity was characterized by mass spectrometry, amino acid composition analysis, and Edman degradation, yielding the sequence GGLKKLGKKLEGVGKRVFKASEKALPVAVGIKALG. Unlike other cecropins, the peptide was not amidated at the C-terminus. Aedes albopictus Cecropin A (AalCecA) is the first cecropin to be described from a mosquito vector of human disease. Consistent with the classification of mosquitoes among the Dipteran suborder Nematocera, AalCecA shares only 36% amino acid identity with cecropins from Drosophila melanogaster and other Cyclorrhaphid flies, whose mature cecropins share 80% to 100% amino acid identity.

Aedes↗
PubMed1998

Reciprocal stimulation between TNF-alpha and nitric oxide may exacerbate CNS inflammation in experimental autoimmune encephalomyelitis.

Nitric oxide (NO) and TNF-alpha are both highly active pleotypic modulators of cell function that are abundantly generated during inflammation. Experiments in animal systems have linked the generation of NO and TNF-alpha to autoimmune pathogenesis, and blockade of either NO or TNF-alpha has been shown to impede disease development. In this study, we show that NO and TNF-alpha can act mutually to stimulate each other's production. While IFN-gamma alone induces NO release from microglia, astrocytes are provoked into significant NO production only by a combination of IFN-gamma and TNF-alpha. Since both TNF-alpha and NO are abundantly generated during T-glial cell interaction, we asked whether and how NO affects TNF-alpha production. Using an in vitro system in which TNF-alpha secretion is induced in MBP-reactive T cells by co-culture with syngeneic astrocytes, we observed that the efficiency of TNF-alpha secretion was markedly increased, in a dose-dependent fashion, by addition of micromolar concentrations of a chemical generator of NO donor, sodium nitroprusside (SNP). Similarly, low concentrations of SNP significantly enhanced the IL-2 dependent growth of MBP-reactive T cells. These results suggest that autoimmune pathogenesis initiated by inflammatory responses within the CNS may result in part from a vicious cycle in which TNF-alpha and NO mutually provoke each other's production.

Animals↗
PubMed1998

Citrullinated myelin basic protein induces experimental autoimmune encephalomyelitis in Lewis rats through a diverse T cell repertoire.

An increased proportion of citrullinated MBP (MBP-C8) occurs in the brains of multiple sclerosis (MS) patients. In this study, MBP-C8 from guinea pig (GP) brains was isolated and found encephalitogenic in Lewis rats upon immunization. An encephalitogenic T cell line selected with MBP-C8 preferentially reacted with MBP-C8 over unmodified MBP. This T cell line responded weakly to the dominant encephalitogenic epitope, GP-MBP peptide 70-88, and did not display restricted TCR beta-chain usage (such as Vbeta88.2). The distinctive features of MBP-C8 were also demonstrated by its ability to reinduce active EAE in 70% of rats which had recovered from unmodified MBP induced EAE. These findings raise the possibility that citrullinated MBP may elicit a different pathogenic T cell repertoire for the recurrent phases of inflammatory demyelination.

Animals↗
PubMed1998

The rluC gene of Escherichia coli codes for a pseudouridine synthase that is solely responsible for synthesis of pseudouridine at positions 955, 2504, and 2580 in 23 S ribosomal RNA.

Escherichia coli ribosomal RNA contains 10 pseudouridines, one in the 16 S RNA and nine in the 23 S RNA. Previously, the gene for the synthase responsible for the 16 S RNA pseudouridine was identified and cloned, as was a gene for a synthase that makes a single pseudouridine in 23 S RNA. The yceC open reading frame of E. coli is one of a set of genes homologous to these previously identified ribosomal RNA pseudouridine synthases. In this work, the gene was cloned, overexpressed, and shown to code for a pseudouridine synthase able to react with in vitro transcripts of 23 S ribosomal RNA. Deletion of the gene and analysis of the 23 S RNA from the deletion strain for the presence of pseudouridine at its nine known sites revealed that this synthase is solely responsible in vivo for the synthesis of three of the nine pseudouridine residues, at positions 955, 2504, and 2580. Therefore, this gene has been renamed rluC. Despite the absence of one-third of the normal complement of pseudouridines, there was no change in the exponential growth rate in either LB or M-9 medium at temperatures ranging from 24 to 42 degrees C. From this work and our previous studies, we have now identified three synthases that account for 50% of the pseudouridines in the E. coli ribosome.

Base Sequence↗
PubMed1998

Cell death mediated by Fas-FasL interaction between glial cells and MBP-reactive T cells.

Apoptosis in T cells that have penetrated into the central nervous system (CNS) may be important for the physiological control of T cells with potentially dangerous reactivities to CNS antigens; such control may be dysfunctional in animals suffering from experimental autoimmune encephalomyelitis (EAE). In this study we examined the expression of Fas and FasL genes both in myelin basic protein (MBP)-reactive T cells and in glial cells and the susceptibility of these cells to death induced by Fas/FasL interaction. Both Fas and FasL gene expression is detectable in glial cells and MBP-reactive T cells. Cell death is not unidirectional: when T cells interact with glial cells death can be induced in the former or in the latter population. The ability to induce death of Fas-expressing cells varies greatly among different lines of MBP-reactive T cells, as does resistance to death induction by cells expressing FasL. Moreover, the ability of T cells both to deliver and to resist death signals is a function of their activation status: T cells freshly activated transmit a stronger apoptotic signal to Fas-positive target cells and are also more resistant to FasL-induced suicide. Soluble form of FasL provides a convenient titratable means of delivering death signals via Fas. However, comparison of the susceptibility of different targets to soluble FasL and to FasL expressed on the surface of a transfected glial line revealed differences, suggesting that signals arising from Fas/FasL interaction may be modulated by additional cell-surface molecules.

Animals↗
PubMed1998

Overexpression of both p185c-erbB2 and p170mdr-1 renders breast cancer cells highly resistant to taxol.

We recently found that overexpression of p185c-erbB2 in c-erbB2 transfected MDA-MB-435 breast cancer cells (435.eB transfectants) confers a 5-9-fold increase in Taxol resistance. To examine whether Taxol resistance is a common phenomenon in other c-erbB2 overexpressing breast cancer cell lines, we tested a panel of human breast cancer cell lines established from different patients and expressing pl85c-erbB2 at different levels for their sensitivity to Taxol and Taxotere, a synthetic taxoid. Higher expression of p185c-erbB2 in these breast cancer cell lines indeed correlated well with resistance to Taxol and Taxotere, and the degree of resistance was about 100-fold that in c-erbB2-overexpressing 435.eB transfectants, demonstrating that these breast cancer cells are highly resistant to Taxol. Since mdr-1-encoded p-glycoprotein (p170mdr-1) has been implicated in Taxol resistance, we next examined the p170mdr-1 levels in these breast cancer cell lines that are highly resistant to Taxol. Higher levels of p170mdr-1 expression were found in several breast cancer cell lines that are highly resistant to Taxol. Since these same breast cancer cell lines also expressed higher levels of p185c-erbB2, we sought to determine the relative contribution of p185c-erbB2 and p170mdr-1 overexpression to Taxol resistance. We first specifically down-regulated cell surface p185c-erbB2 using anti-p185c-erbB2 monoclonal antibodies and assayed sensitivity of these cells to Taxol. We next specifically inactivated p170mdr-1 function using p170mdr-1 blockers (thioridazine or verapamil) and again assayed Taxol sensitivity. Both p185c-erbB2 down-regulation and p170mdr-1 blockade significantly sensitized the breast cancer cell lines to Taxol. The results indicate that overexpression of either p185c-erbB2 or p170mdr-1 renders human breast cancer cells resistant to Taxol. Furthermore, p185c-erbB2 synergizes with p170mdr-1 conferring higher degrees of Taxol resistance. Finally, combination therapy (down-regulation of p185c-erbB2 plus blocking p170mdr-1 plus administration of Taxol) may be beneficial to breast cancer patients whose tumors express high levels of both p185c-erbB2 and p170mdr-1.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Effect of DNA secondary structure on human telomerase activity.

Telomeres are specialized DNA-protein complexes located at the chromosome ends. The guanine-rich telomeric sequences have the ability to form G-quadruplex structures under physiological ionic conditions in vitro. Human telomeres are maintained through addition of TTAGGG repeats by the enzyme telomerase. To determine a correlation between DNA secondary structure and human telomerase, telomerase activity in the presence of various metal cations was monitored. Telomerase synthesized a larger proportion of products corresponding to four, five, eight, and nine full repeats of TTAGGG in 100 mM K+ and to a lesser extent in 100 mM Na+ when a d(TTAGGG)3 input primer was used. A more even product distribution was observed when the reaction mixture contained no added Na+ or K+. Increasing concentrations of Cs+ resulted in a loss of processivity but not in the distinct manner observed in K+. When the input primer contained 7-deaza-dG, the product distribution resembled that of reactions without K+ even in the presence of 100 mM K+. Native polyacrylamide gel electrophoresis indicated that d(TTAGGG)4, d(TTAGGG)5, d(TTAGGG)8, and d(TTAGGG)9 formed compact structures in the presence of K+. The oligonucleotide d(TTAGGG)4 had a UV spectrum characteristic of that of the G-quadruplex only in the presence of K+ and Na+. A reasonable explanation for these results is that four, five, eight, and nine repeats of TTAGGG form DNA secondary structures which promote dissociation of the primer from telomerase. This suggests that telomerase activity in cells can be modulated by the secondary structure of the DNA template. These findings are of probable relevance to the concept of telomerase as a therapeutic target for drug design.

Cesium↗
PubMed1998

Expression of the rat adrenomedullin receptor or a putative human adrenomedullin receptor does not correlate with adrenomedullin binding or functional response.

There has been considerable difficulty in defining distinct adrenomedullin (AM) binding sites and function in vivo. However, a rat adrenomedullin receptor (rAMR) and a putative human adrenomedullin receptor (hAMR) have recently been reported. We attempted to confirm and extend the pharmacological characterization of these cloned receptors. COS-7 cells transfected with rAMR or epitope tagged rAMR display abundant rAMR mRNA expression and cell-surface receptor localization. Specific 125I-AM binding is detected in transfected cells; however, similar levels of binding are also detected in cells transfected with vector DNA alone. This AM binding site fails to mediate any changes in cAMP in response to AM. In contrast, Swiss 3T3 cells, expressing specific endogenous AM receptors, display AM binding and functional cAMP responses. Transfection studies performed with the putative hAMR yield similar results. These data suggest that the proposed rAMR and hAMR do not represent authentic adrenomedullin receptors.

3T3 Cells↗
PubMed1998

[The relationship between angiotensin II type 1 receptor gene and coronary heart disease, hypertension and diabetes mellitus in Chinese].

OBJECTIVE: To ascertain whether the angiotensin II type 1 receptor (AGTR1) gene is associated with coronary heart disease (CHD), hypertension (HTN) and non-insulin-dependent diabetes mellitus (NIDDM) in Chinese and to observe the interaction of AGTR1 gene and angiotensin I converting enzyme(ACE) gene to the development of the associated disease in Chinese. METHODS: 270 cases of Chinese with CHD, HTN or NIDDM in single or in different combinations were genotyped for the A1166C variation at the 3'-untranslated region of AGTR1 gene by using PCR/Dde I restriction endonuclease digestion. And also, the insertion/deletion polymorphism at the intron 16 of ACE gene was typed by PCR. RESULTS: (1) AGTR1 gene was associated with CHD (logistic regression analysis, P=0.02), but not with HTN and NIDDM; (2) The attributable risk of AGTR1 gene for CHD was 12.5% and the odds ratio was 4.55. Both AGTR1 gene and ACE gene were independent variation factors for CHD (P=0.032 and P=0.002 respectively); (3) The contribution of AGTR1 gene to the development of CHD was independent of total body adiposity, blood lipid profile and blood pressure; (4) The analysis of individual combined genotypes of AGTR1 gene and ACE gene showed significant difference in frequency distribution between patients with CHD and controls. CONCLUSION: AGTR1 gene contributes to the development of CHD and is an independent risk factor for CHD in Chinese.

Adult↗
PubMed1998

Cloning, expression, and purification of a catalytic fragment of Moloney murine leukemia virus reverse transcriptase: crystallization of nucleic acid complexes.

Reverse transcriptase is an essential retroviral enzyme that uses RNA- and DNA-directed DNA polymerase activities as well as an RNaseH activity to synthesize a double-stranded DNA copy of the single-stranded RNA genome. In an effort to obtain high-resolution structural information regarding the polymerase active site of reverse transcriptase, we have pursued studies on a catalytic fragment from Moloney murine leukemia virus reverse transcriptase. DNA encoding the catalytic fragment, defined originally by limited proteolytic digestion, has been cloned, and the protein has been expressed and purified from Escherichia coli. The fragment obtained by limited proteolytic digestion and the bacterially expressed fragnment retain polymerase activity. Crystallization studies involving nucleic acid complexes with a catalytic fragment from both sources are reported, including variables screened to improve crystals and cryocooling. Three crystal forms of catalytic fragment-nucleic acid complexes have been characterized, which all contain at least two protein molecules in the asymmetric unit. As isolated, the catalytic fragment is monomeric. This analysis indicates that the enzyme dimerizes in the presence of nucleic acid.

Base Sequence↗
PubMed1998

Coexistence of low and high grade squamous intraepithelial lesions of the cervix: morphologic progression or multiple papillomaviruses?

BACKGROUND: The diagnosis of both low (LSIL) and high (HSIL) grade squamous intraepithelial lesions in the same cervical specimen may reflect classification variation, morphologic progression in situ, and, conceivably, different HPV infections. We addressed these possibilities in cervical specimens previously diagnosed as containing both LSIL (condyloma/CIN1) and HSIL (CIN2/3). METHODS: All cases with a histologic diagnosis of LSIL and HSIL from 1994-1996 were reviewed. On review, lesions were scored as (1) no significant variation in lesion grade (classification discrepancies) and showing a (2) one (CIN1-2) or (3) two (CIN1-3) grade shift in the same case. In cases in which a one or two grade shift was confirmed, low (CIN1) and high (CIN2-3) grade foci were microdissected and extracted DNA analyzed for HPV by PCR and RFLP analysis. RESULTS: Of 98 cases available for review, 58 (59%) did not exhibit significant variation in grade (classification discrepancy), and 40 (41%) showed a one (25) or two (15) grade shift. Of the latter group both LSIL and HSIL foci were HPV(+) in 26 (65. 0%). The same HPV was present in both LSIL and HSIL foci in 15/15 lesions with a one grade shift (CIN1-2). In contrast, a significantly higher proportion of lesions with a two grade shift (CIN1-3) contained two different HPV types (4/11 vs 0/15; P = 0.01). Combinations of HPVs in the low/high grade foci, respectively, included HPV 11/16 (1), 11/16 + 18 (1), and HPV39/16 (2). CONCLUSIONS: Lesions containing LSIL and HSIL which span two grades (CIN1 and CIN2) most likely represent morphologic progression in a single infection. Lesions containing CIN1 and CIN 3 may be attributed to both lesion progression and two coincident infections; the latter sometimes present in the same histologic section. The latter phenomenon has implications for both the diagnosis of CIN and interpretation of "morphologic progression" from very low to high grade in the same case.

DNA Primers↗
PubMed1998

Isolation and comparison of natural and recombinant human CENP-A autoantigen.

Anticentromere antibodies (ACA) are associated with systemic sclerosis (scleroderma) patients exhibiting the more benign or so called limited manifestation of the disease (lSSc). ACA reactivity is directed against multiple polypeptide targets, the smallest of which is designated CENP-A. CENP-A is not an abundant cellular constituent; therefore to maximize recovery, we developed a protocol with a minimum of steps to isolate CENP-A from a human cell line. The trace cellular amount of this protein clearly dictated the production of its recombinant counterpart to facilitate determination of the role of the CENP-A antigen in scleroderma pathogenesis. Here we describe the eukaryotic expression of CENP-A cDNA using baculovirus-mediated infection of insect cells. The non-fusion recombinant protein spans the natural residues of the human CENP-A protein and rCENP-A followed the same chromotographic sequence for purification as did the natural source. The availability of the bona fide antigen provided a critical standard upon which to document authenticity of the recombinant polypeptide. The two forms of this antigen have been compared and shown to exhibit similar physical and antigenic properties.

Animals↗