Biomedical subjects

X Xu

Publications and source records attributed to X Xu.

At least 289 records · Page 16Linked to original sources

PubMed2000

Effects of semen plasma from different fractions of individual ejaculates on IVF in pigs.

We applied IVM/IVF techniques to investigate effects of preincubation of sperm with different fractions of semen plasma harvested from fresh ejaculates on in vitro penetration and fertilization of in vitro matured oocytes. Three fractions of semen plasma were separated from the complete ejaculate of three Landrace boars and used to coincubate sperm obtained from the first sperm-rich fraction of the same ejaculates. After 14 to 16 h coincubation at room temperature, sperm were preincubated in capacitation medium and then inseminated into fertilization medium containing porcine oocytes matured in vitro. The semen plasma used for coincubation affected penetration rate (P < 0.001); Sperm coincubated with Fraction 1 semen plasma had a higher penetration rate compared with sperm coincubated with Fraction 2 (P < 0.05), but not with Fraction 3. Boar affected male pronucleus formation rates after insemination (P < 0.05), but no difference among boars was found in monospermy rate, average number of sperm penetrating into each fertilized oocyte, or the average number of sperm attached. No boar by fraction interaction was found for any parameters studied.

Animals↗

PubMed2000

P38 MAPK, but not p42/p44 MAPK mediated inducible nitric oxide synthase expression in C6 glioma cells.

Recently mitogen-activated protein kinase (MAPK) has been reported to play an important role in phosphorylation cascades governing cell growth and protein expression in numerous cell types. In order to explore the signaling mechanism by which inducible nitric oxide synthase (iNOS) is regulated in C6 glioma cells, we investigated the role of MAPK in iNOS expression by using the specific MAPK inhibitors. First the induction of nitric oxide by lipopolysaccharide (LPS), tumor necrosis factor alpha (TNFalpha), interferon gamma (IFNgamma), alone or their combination, was studied in C6 glioma cells. Administration of LPS, TNFalpha, or IFNgamma alone had no detectable stimulatory effect on the production of nitric oxide (NO). However, combination of the three factors elicited a significant elevation of NO level in C6 cell culture medium. Subsequently pretreatment of C6 cells with a specific inhibitor of p38 MAPK, SB202190, resulted in a dose-dependent inhibition of NO production and iNOS expression, but PD98059, an inhibitor of p42/p44 MAPK activation, had no effect. These data suggest that p38 MAPK mediates iNOS expression in C6 glioma cells, but p42/p44 MAPK is not involved in this process.

Cytokines↗

PubMed2000

Novel human esophagus-specific gene c1orf10: cDNA cloning, gene structure, and frequent loss of expression in esophageal cancer.

We have identified a novel human gene, designated C1orf10, using modified differential display PCR. The C1orf10 gene, which spans 5 kb in length, is composed of three exons. The deduced protein contains 495 amino acids with one transmembrane domain. The amino acid sequence of C1orf10 is characterized by the presence of a calcium-binding motif of about 90 amino acids at its N-terminal and a conserved consecutive repeat sequence of 60 amino acids that was identified previously only in bacterial ice nucleation proteins. In normal adult tissues, C1orf10 is highly expressed only in the esophagus and was undetectable in a total of 15 other tissues examined, suggesting its important role in esophageal cells. The expression of C1orf10 is either dramatically reduced or absent in esophageal cancer cell lines (3/3) as well as primary esophageal cancer tissues (35/37) compared with the corresponding normal esophageal mucosa. Using a radiation hybrid panel, C1orf10 was found to be located on chromosome 1q21. These findings suggest that expression of C1orf10 is unique to esophageal cells and that loss of its expression may play a role in the development of esophageal cancer.

Adult↗

PubMed2000

A liquid chromatography-mass spectrometry method for the quantification of bioavailability and bioconversion of beta-carotene to retinol in humans.

A method based on high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (APCI LC-MS) was developed for the quantification of the bioavailability of retinyl palmitate and beta-carotene and the bioconversion of beta-carotene to retinol in humans. Following oral administration of [8,9,10,11,12,13,14,15,19,20-13C10]-retinyl palmitate and [12,13,14,15,20,12',13',14',15',20'-13C10]-beta-carotene at physiological doses to children between 8 and 11 years of age, blood samples were drawn and serum was prepared. Retinol and beta-carotene were extracted from 0.2- and 1.0-mL serum samples, respectively, and analyzed using reversed-phase HPLC with a C30 column interfaced to an APCI mass spectrometer. Unlike other LC-MS assays for carotenoids, no additional purification steps were necessary, nor was any derivatization of retinol or beta-carotene required. APCI LC-MS showed a linear detector response for beta-carotene over 4 orders of magnitude. Using selected ion monitoring to record the elution profile of protonated circulating beta-carotene at m/z 537 and [13C10]-beta-carotene at m/z 547, the limit of detection was determined to be 0.5 pmol injected on-column. To assess the ratio of labeled to unlabeled retinol, selected ion monitoring was carried out at m/z 269, 274, and 279. These abundant fragment ions corresponded to the loss of water from the protonated molecule of circulating retinol, [13C5]-retinol (metabolically formed from orally administered [13C10]-beta-carotene), and [13C10]-retinol (formed by hydrolysis of [13C10]-retinyl palmitate). The ratios of labeled to unlabeled retinol and the ratio of labeled to unlabeled beta-carotene were calculated. Combined with standard HPLC measurement of beta-carotene and retinol concentration and a mathematical model, these results showed that this simple LC-MS method can be used to quantify beta-carotene bioavailability and its bioconversion to retinol at physiologically relevant doses.

Biological Availability↗

PubMed2000

Genetic susceptibility to benzene and shortened gestation: evidence of gene-environment interaction.

This study investigated whether the association between low level benzene exposure and shortened gestation is modified by two susceptibility genes, CYP1A1 and GSTT1. This report includes 542 (302 nonexposed, 240 benzene-exposed) nonsmoking and nondrinking mothers of singleton live births at Beijing Yanshan Petrochemical Corporation between June 1995 and June 1997. Epidemiologic and clinical data and blood samples were obtained from mothers. Multiple linear regression models were used to estimate the associations of benzene exposure and genetic susceptibility with gestational age, adjusting for maternal age, education, parity, stress, passive smoking, prepregnancy weight and height, and infant's sex. Without consideration of genotype, benzene exposure was associated with a decrease in mean gestational age of 0.29 (standard error (SE), 0.12) week. When stratified by the maternal CYP1A1 genotype, the estimated decrease was 0.54 (SE, 0.12) week for the AA group, which was significantly greater (p = 0.003) than that for the Aa/aa group, which showed no decrease in gestational age. When both CYP1A1 and GSTT1 were considered, the greatest decrease was found among exposed mothers with the CYP1A1 AA-GSTT1 absent group (0.79 (SE, 0.25) week) and the CYP1A1 AA-GSTT1 present group (0.50 (SE, 0.22) week). Among the nonexposed, genetic susceptibility alone did not confer a significant adverse effect. This study provides evidence of gene-environment interaction and supports further assessment of the role of genetic susceptibility in the evaluation of reproductive toxins.

Adult↗

PubMed2000

Stimulation of MCF-7 cell proliferation by low concentrations of Chinese domestic polychlorinated biphenyls.

The simple and sensitive in vitro MCF-7 human breast cancer cell proliferation assay was used to examine the proliferation abilities of two Chinese commercial polychlorinated biphenyl (PCB) mixtures made in the 1960s. Chinese PCB3 and Chinese PCB5 were compared with 17 beta-estradiol (E2). All of the positive activities of these types of Chinese PCBs were significantly different compared to controls with respect to MCF-7 cell doubling time. At lower levels of 7.8 pg/ml and 182 pg/ml, the Chinese PCB3 showed 94% and 86% of relative proliferation effects compared to 17 beta-estradiol, respectively. Chinese PCB5, also showed higher cell proliferation activity at lower level of 8.3 pg/ml, with relative proliferation effect as high as 107% in comparison to 17 beta-estradiol. Thus, both PCBs seem to be different from corresponding Aroclor mixtures. However, Chinese PCBs did not express cell proliferation effects at higher levels of 9.1 ng/ml for Chinese PCB3 and 166 pg/ml and 8.3 ng/ml for Chinese PCB3. This may be due to cytotoxicity and/or antiestrogenic compounds in the mixtures.

Breast Neoplasms↗

PubMed2000

A sensitivity analysis on the atmospheric transformation and deposition of mercury in north-eastern USA

This paper presents the results of a sensitivity analysis on the factors that affect dry and wet deposition of atmospheric mercury (Hg), using a regional scale air quality model. Simulations were conducted for the north-eastern USA during a summer week and a winter week in 1997. Simulation results for the summer week and the winter week in general showed similar responses to changes in emission, environmental conditions, and alternative chemical mechanisms. Reduction of the ambient concentrations of soot or ozone was shown to reduce the wet deposition of Hg. When averaged over the summer and the winter week, the total deposition to the simulation domain would be reduced by 26% by reducing Hg emission from anthropogenic sources within the domain by 50%. For individual grids, however, only locations near local sources obtained noticeable reductions in ambient concentration and wet deposition due to the influence of re-emission from the natural surfaces and regional/global scale transport. The reduction in deposition would reach 36% if all Hg(II) emitted from anthropogenic sources were attached to particles. The total deposition was predicted to decrease by 22% when the gas phase Hg(II)-Hg(p) partitioning was included in the model. Only small changes in total deposition were observed by including the gas-phase ozone-Hg(0), reaction and the aqueous phase chlorine-Hg(0), reaction, and by lowering ambient concentrations of Hg(II) and Hg(p) at the upper lateral boundaries. During the summer week, Hg(II) deposition contributed 40% or more to the total deposition. The contribution increased to 70% in the winter week.

Journal Article↗

PubMed2000

Sound velocity measurements of nuclear-ordered solid 3He along the melting curve.

We have measured the temperature dependence of the 10.98 MHz longitudinal sound velocity of solid 3He in the nuclear-ordered U2D2 phase and in the paramagnetic phase along the melting curve. The temperature dependence of the sound was attributed to the contributions from the nuclear spin system and the molar volume change along the melting curve. The sound velocity increased with temperature as T4 in the U2D2 phase and the sound anisotropy due to the exchange interaction was found to be about 10% among single-domain crystals investigated. The average value of the Gruneisen constant of the spin wave velocity in the ordered phase was gamma(c) = 16 and is compared to the calculated value of the multiple-exchange model.

Journal Article↗

PubMed2000

Trafficking of human ADAM 12-L: retention in the trans-Golgi network.

We have investigated the trafficking of the membrane-anchored form of human ADAM 12 (ADAM 12-L) fused to a green fluorescence protein tag. Subcellular localization of the protein in transiently transfected cells was determined by fluorescence microscopy and trypsin sensitivity. Full-length ADAM 12-L was retained in a perinuclear compartment, which was shown to be the trans-Golgi network. In contrast, ADAM 12-L lacking the cytoplasmic domain reached the cell surface. Based on analysis of deletions and mutations of the cytoplasmic tail of ADAM 12-L, the retention signal is comprised of both the cytoplasmic and transmembrane domains, but not the Src homology 3 domain (SH3) binding sites. These results raise the possibility that a trafficking checkpoint in the trans-Golgi network is one of the cellular mechanisms for regulation of ADAM 12-L function, by allowing a rapid release of ADAM 12-L to the cell surface under specific stimuli.

ADAM Proteins↗

PubMed2000

A unified Haseman-Elston method for testing linkage with quantitative traits.

The Haseman and Elston (H-E) method uses a simple linear regression to model the squared trait difference of sib pairs with the shared allele identical by descent (IBD) at marker locus for linkage testing. Under this setting, the squared mean-corrected trait sum is also linearly related to the IBD sharing. However, the resulting slope estimate for either model is not efficient. In this report, we propose a simple linkage test that optimally uses information from the estimates of both models. We also demonstrate that the new test is more powerful than both the traditional one and the recently revisited H-E methods.

Alleles↗

PubMed2000

Targeting and therapy of carcinoembryonic antigen-expressing tumors in transgenic mice with an antibody-interleukin 2 fusion protein.

The purpose of this study was to engineer a bivalent single-chain anticarcinoembryonic antigen (CEA) antibody and an interleukin 2 (IL-2) fusion protein derivative for selective tumor targeting of cytokines. The variable domains of a high affinity anti-CEA antibody, T84.66, were used to form a single-gene-encoded antibody [single-chain variable fragment joined to the crystallizable fragment, Fc (scFvFc)]. The fusion protein (scFvFc.IL-2) consisted of mouse IL-2-fused to the COOH-terminal end of the scFvFc. The engineered proteins were assembled as complete molecules and were similar to the intact anti-CEA monoclonal antibody (Mab) in antigen-binding properties. Based on IL-2 content of the fusion protein, its ability to support proliferation of CTLL-2 cells was identical with that of IL-2. Despite a molecular size similar to that of the intact Mab, the blood clearance of the fusion protein was markedly faster than that of the intact Mab or scFvFc. Incubation of radiolabeled scFvFc.IL-2 but not the intact or scFvFc antibodies in mouse serum was accompanied by the appearance of complexes, suggesting that the latter may contribute to the accelerated clearance of the fusion protein. Biodistribution and tumor targeting studies were carried out in CEA-transgenic mice bearing CEA-positive murine tumors as well as the antigen-negative parental tumor. The bivalent anti-CEA scFvFc had tumor localization properties similar to those of the intact Mab. Although fusion of IL-2 to the COOH-terminal end of the bivalent scFvFc altered its pharmacokinetic properties, the fusion antibody was able to target tumors specifically. Maximum uptake of the intact Mab, scFvFc, and scFvFc.IL-2 in CEA-positive tumors was 29.3 +/- 5.0, 19.5 +/- 2.1, and 6.6 +/- 0.9% injected dose/g, respectively. Maximum tumor localization ratios (CEA-positive/CEA-negative tumor) were similar for all three antibody types (4.6-6.0), demonstrating the antigen specificity of the tumor targeting. Significant antigen-specific targeting to CEA-positive normal tissues of transgenic mice was not observed. Although the tumor-targeting properties of the fusion protein were low, the growth of CEA-expressing (P = 0.01) but not antigen-irrelevant (P = 0.22) syngeneic tumor cells was inhibited after treatment of transgenic mice with the anti-CEA-IL-2 antibody. Therapy of CEA-expressing tumors was improved after i.v. administration of the fusion protein (P = 0.0001). These studies indicate that anti-CEA antibody-directed cytokine targeting may offer an effective treatment for CEA-expressing carcinomas. The availability of an immunocompetent CEA transgenic mouse model will also help to determine the immunotherapeutic properties of these fusion proteins.

Animals↗

PubMed2000

A Pro250Arg substitution in mouse Fgfr1 causes increased expression of Cbfa1 and premature fusion of calvarial sutures.

Pfeiffer syndrome is a classic form of craniosynostosis that is caused by a proline-->arginine substitution at amino acid 252 (Pro252Arg) in fibroblast growth factor receptor 1 (FGFR1). Here we show that mice carrying a Pro250Arg mutation in Fgfr1, which is orthologous to the Pfeiffer syndrome mutation in humans, exhibit anterio-posteriorly shortened, laterally widened and vertically heightened neurocraniums. Analysis of the posterior and anterior frontal, sagittal and coronal sutures of early post-natal mutant mice revealed premature fusion. The sutures of mutant mice had accelerated osteoblast proliferation and increased expression of genes related to osteoblast differentiation, suggesting that bone formation at the sutures is locally increased in Pfeiffer syndrome. Of note, dramatically increased expression of core-binding transcription factor alpha subunit type 1 (Cbfa1) accompanied premature fusion, suggesting that Cbfa1 may be a downstream target of Fgf/Fgfr1 signals. This was confirmed in vitro, where we demonstrate that transfection with wild-type or mutant Fgfr1 induces Cbfa1 expression. The induced expression was also observed using Fgf ligands (Fgf2 and Fgf8). These studies provide direct genetic evidence that the Pro252Arg mutation in FGFR1 causes human Pfeiffer syndrome and uncovers a molecular mechanism in which Fgf/Fgfr1 signals regulate intramembraneous bone formation by modulating Cbfa1 expression.

Acrocephalosyndactylia↗

PubMed2000

Selective cyclooxygenase-2 inhibitors: heteroaryl modified 1,2-diarylimidazoles are potent, orally active antiinflammatory agents.

A series of heteroaryl modified 1,2-diarylimidazoles has been synthesized and found to be potent and highly selective (1000-9000-fold) inhibitors of the human COX-2. 3-Pyridyl derived COX-2 selective inhibitor (25) exhibited excellent activity in acute (carrageenan induced paw edema, ED(50) = 5.4 mg/kg) and chronic (adjuvant induced arthritis, ED(50) = 0.25 mg/kg) models of inflammation. The relatively long half-life of 25 in rat and dog prompted investigation of the pyridyl and other heteroaromatic systems containing potential metabolic functionalities. A number of substituted pyridyl and thiazole containing compounds (e.g., 44, 46, 54, 76, and 78) demonstrated excellent oral activity in every efficacy model evaluated. Several orally active diarylimidazoles exhibited desirable pharmacokinetics profiles and showed no GI toxicity in the rat up to 100 mg/kg in both acute and chronic models. The paper describes facile and practical syntheses of the targeted diarylimidazoles. The structure-activity relationships and antiinflammatory properties of a series of diarylimidazoles are discussed.

Administration, Oral↗

PubMed2000

Changes of reliability and efficiency of micronucleus bioassay in Vicia faba after exposure to metal contamination for several generations.

Mitotic root micronucleus (MCN) frequency in Vicia faba as a bioassay, is primarily based on the extent of the sentinel Vicia response in terms of cytogenetic damage quantitatively or qualitatively to indicate the presence of mutagenic contaminants. This paper describes an investigation designed to assess changes in MCN frequencies of Vicia faba from three generation plants obtained, respectively from a reference site (RS) and a metal-contaminated experimental field (EF) in the bioassay of mutagenic Cd(2+) and NaN(3). The background value, dose-response to Cd(2+) and to NaN(3) in three generation (F(1), F(2) and F(3)) plants of the EF and the initial (F(0)) plants were determined in terms of MCN frequencies. With more generations of growing Vicia plants in the EF, a higher background value of MCN frequency, a lower slope value in the regression equation, a smaller ratio of MCN frequency between the control and treatment in the same generation and larger perturbation values were observed. This denotes that the decreased reliability and efficiency are represented in Vicia plants from the EF if the plants are used as sentinels in the bioassay of mutagenic Cd(2+) and NaN(3). It was concluded that the Vicia MCN bioassay should be used as an endpoint biomarker acceptable in biomonitoring environmental mutagens when the sentinel plants were collected from clean areas. Because no place is absolutely without pollutants, it is suggested that several seed stock centers should be established for providing sentinel Vicia if Vicia MCN bioassay is used as a biomarker to identify the global environmental status.

Journal Article↗

PubMed2000

Combinatorial diversification of indinavir: in vivo mixture dosing of an HIV protease inhibitor library.

An efficient combination solution-phase/solid-phase route enabling the diversification of the P1', P2', and P3 subsites of indinavir has been established. The synthetic sequence can facilitate the rapid generation of HIV protease inhibitors possessing more favorable pharmacokinetic properties as well as enhanced potencies. Multiple compound dosing in vivo may also accelerate the identification of potential drug candidates.

Animals↗

PubMed2000

Unique carboxyl-terminal sequences of wild type and alternatively spliced variant forms of transforming growth factor-alpha precursors mediate specific interactions with ErbB4 and ErbB2.

We have previously reported that the human transforming growth factor-alpha (TGF-alpha) gene encodes three forms of TGF-alpha precursors, designated wild type (WT), variant I (VaI), and variant II (VaII), derived from alternative splicing. The two carboxyl-terminal valine residues of WT are replaced by 5 (GCRLY) or 4 (ATLG) amino acids in VaI or VaII, respectively. When overexpressed in Chinese hamster ovary (CHO) cells, VaI and ValI, but not WT, support autonomous growth. We detected tyrosine phosphorylation of ErbB2 in the absence of serum, in CHO cells expressing WT, VaI, or VaII, but not in mock transfectants. These observations prompted us to investigate possible interactions between the ErbBs and the TGF-alpha precursors in CHO cells. All TGF-alpha precursors were found to co-immunoprecipitate with the ErbBs, but with different specificity. WT co-immunoprecipitated with ErbB4, but not with ErbB1, ErbB2, or ErbB3. VaI and VaII co-immunoprecipitated with ErbB2, but not with ErbB1, ErbB3, or ErbB4. Confocal fluorescent microscopy analysis demonstrated that WT, VaI, and VaII all distribute equally to the cell surface while, as expected, a WT mutant lacking the two C-terminal valine residues does not. Point and deletion mutants involving the unique carboxyl-terminal residues of WT, VaI and VaII, indicated that the interactions between the three TGF-alpha precursors and the ErbBs were mediated by their carboxyl-terminal regions, which constitute distinct protein-binding motifs. A chimera of the intracellular domain of WT TGF-alpha linked to exogenous transmembrane and extracellular domains retained both the cell surface distribution and the specific interaction with ErbB4 of full-length WT, confirming that this interaction is mediated by the C-terminus of the TGF-alpha precursor. While interactions of WT and variant TGF-alpha with the ErbBs all result in ErbB2 activation, they produce different biological consequences, suggesting that the various TGF-alpha precursors differentially modulate ErbB signaling.

Alternative Splicing↗

PubMed2000

Intracerebroventricular infusion of nerve growth factor induces pain-like response in rats.

New strategies have recently been developed where infusion of neurotrophic factors into the brain can rescue different neuronal populations. However, negative side effects have been observed in clinical trials infusing nerve growth factor (NGF) into the lateral ventricle in man, namely pain. Little is known about pain behavior in animals after intracerebroventricular (i.c.v.) neurotrophic injections. Thus, we have examined the effects of i.c.v. infusion of NGF for 2 weeks on the behavioral response of rats to mechanical, cold and heat stimulation. Seven micrograms/day of NGF elicited a significant decrease in vocalization threshold to mechanical stimulation and a significantly increased response to cold and heat stimuli as compared with control. The concentration of NGF in cerebrospinal fluid (CSF) was significantly increased as compared with non-allodynic rats. The enhanced responses to mechanical and heat, but not to cold, stimulation were significantly reduced by CP-99994, a selective antagonist to tachykinin NK-1 receptors. When NGF was infused into the brain parenchyma (striatum, cortex and septum) no allodynic nor hyperalgesic responses could be detected. These results indicate that in rats i.c.v. but not intraparenchymal infusion of NGF induce mechanical and cold allodynia as well as heat hyperalgesia, which is mediated, at least in part, by activation of NK-1 receptors.

Animals↗

PubMed2000

Heat transport in turbulent rayleigh-Benard convection

We present measurements of the Nusselt number N as a function of the Rayleigh number R in cylindrical cells with aspect ratios 0. 5</=gamma identical withD/d</=12.8 ( D is the diameter and d is the height). We used acetone with a Prandtl number sigma = 4.0 for 10(5) less, similarR less, similar4x10(10). A fit of a power law N = N(0)R(gamma(eff)) over limited ranges of R yielded values of gamma(eff) from 0.275 near R = 10(7) to 0.300 near R = 10(10). The data are inconsistent with a single power law for N(R). For R>10(7) they are consistent with N = asigma-1/12R1/4+bsigma-1/7R3/7 as proposed by Grossmann and Lohse for sigma greater, similar2.

Journal Article↗