Search PubMed⌕ Search

Biomedical subjects

W Li

Publications and source records attributed to W Li.

At least 109 records · Page 6Linked to original sources
PubMed2003

Reduced birth defects caused by maternal immune stimulation may involve increased expression of growth promoting genes and cytokine GM-CSF in the spleen of diabetic ICR mice.

Maternal immune stimulation in mice decreases fetal abnormalities caused by diverse etiologies. Growth factors produced by activated immune cells were proposed to be key mediators that may exert their effects on placenta or embryo. Diabetes disrupts the secretion of cytokines, which may associate with diabetic embryopathy. Three different methods of maternal immune stimulation that result in approximately equal reduction of diabetic embryopathy were used in the present studies: footpad injection with complete Freund's adjuvant (CFA), intraperitoneal (i.p.) injection with granulocyte-macrophage colony-stimulating factor (GM-CSF), or i.p. injection with interferon-gamma (IFN-gamma). A gene microarray was then used to examine expression of a selected gene panel in splenic leukocytes. We hypothesized that maternal immune stimulation may act by overcoming altered gene expression patterns of immune cells in the diabetic mice, which partially mitigates the teratogenic effect of diabetes. It further seemed likely that a shared profile of splenic gene expression changes induced by the different immune stimulation procedures may be identified and related to reduced teratogenesis. The three procedures produced a common altered gene expression profile. Significantly affected genes included apoptotic and anti-apoptotic genes, and genes controlling cellular proliferation, and likely reflect a state of immune activation. The GM-CSF gene was up-regulated by all three immune stimulation procedures. The protein product of this gene regulates placental development, and was recently associated with reduced cleft palate in immune-stimulated pregnant mice after exposure to urethane. These data suggest that further studies of GM-CSF as mediator of reduced birth defects in teratogen-challenged, immune-stimulated mice are warranted.

Analysis of Variance↗
PubMed2003

Utility of peptide-protein affinity complexes in proteomics: identification of interaction partners of a tumor suppressor peptide.

We used a N-biotinylated peptide analog of the C-terminal domain of the tumor suppressor protein, p21cip1/waf1 to elucidate peptide/protein interacting partners. The C-terminal domain of p21cip1/waf1 protein spanning 141-160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell-cycle control. This C-terminal 20-mer efficiently extracts PCNA in the presence of a variety of N- or C-terminally attached affinity tags. Using difference silver stained 2D gels combined with in-gel tryptic digests, we identified the difference spots using MALDI-TOF mass spectrometry-based peptide mass fingerprinting followed by a database search using PROFOUND against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor-1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion-trap LC-MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using SEQUEST confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90alpha, HSP40 and T-complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21cip1/waf1 protein. The use of N-biotinylated peptide derived from the C-terminal domain of p21cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets.

Amino Acid Sequence↗
PubMed2003

An investigation of the specificity of induced anti-pig antibodies in baboons.

AIM: To provide information on the specificity of induced anti-pig antibodies (Abs) in baboons after exposure and sensitization to pig antigens. MATERIALS AND METHODS: Baboons (n=7) received either porcine mobilized peripheral blood progenitor cells (n=3), kidney (n=3) or heart (n=1) transplants. After rejection of these cells or organs, pre- and post-rejection sera were analyzed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry to detect and measure anti-Galactosealpha1,3Galactose (Gal) and anti-non-Gal Abs. To study the anti-non-Gal carbohydrate response, the sera were incubated with pig red blood cells pretreated with alpha-galactosidase (to remove Gal) and three other exoglycosidases to remove other potential oligosaccharide epitopes, and studied by flow cytometry. To study the anti-swine leukocyte antigen (SLA) response, non-Gal Abs from two baboons sensitized with kidneys from inbred miniature swine of dd or aa haplotype, respectively, were adsorbed on cells of aa, cc, or dd haplotypes, and binding to aa, cc or dd cells was measured by flow cytometry. Cytotoxicity of anti-non-Gal Abs was tested in vitro by a complement-mediated cytotoxicity assay, using pig cells as targets. RESULTS: In pre-transplant and pre-rejection sera, anti-Gal Abs were detected, but anti-non-Gal Abs were either absent or at minimal levels. After exposure to pig antigens, baboons developed induced anti-Gal and anti-non-Gal Abs. No anti-non-Gal Abs directed to the tested carbohydrate epitopes could be detected. Anti-non-Gal Abs showed minor evidence of specific SLA haplotype reactivity, suggesting that the major Ab response was to pan-pig determinants. Anti-non-Gal Abs showed a low level of complement-mediated lysis of pig cells in vitro. CONCLUSIONS: In this limited study, no Ab response to non-Gal carbohydrates was observed, and anti-SLA specificity was minor, indicating that most induced anti-non-Gal Ab was directed against non-specific pig proteins, including SLA-epitopes.

Animals↗
PubMed2003

Anti-inflammatory effects of high-dose montelukast in an animal model of acute asthma.

BACKGROUND: Asthmatic inflammation is mediated by a network of cytokines, chemokines and adhesion molecules. Corticosteroids are the only effective agents available to control asthmatic inflammation. We investigated the effect of high-dose montelukast (MK), a selective cysteinyl leukotriene receptor 1 antagonist, on mediators of airway inflammation. OBJECTIVE: The aim of this study was to determine the effect of a 3-day course of high-dose MK on mediators of airway inflammation induced by a single allergen challenge in sensitized mice. METHODS: Ovalbumin (OVA)-sensitized BALB/c mice were treated with 25 mg/kg of MK or saline intravenously for 3 days. On the third day, a single inhalation challenge with OVA was given. Cellular infiltration was assessed in the bronchoalveolar lavage (BAL) and in the lung. Expression of IL-4, IL-5, IL-13 and eotaxin in the BAL, and the lung was determined. Serum IL-5 and total IgE was measured. IL-5 and eotaxin mRNA expression in the lung was determined. Finally, eotaxin and VACM-1 expression in the lung was assessed by immunohistochemistry. RESULTS: MK reduced the number of eosinophils in the BAL by > 90%. There was also significant reduction in IL-5 in the BAL, lung and the serum, and IL-5 mRNA expression in the lung. IL-4 level in the lung and BAL, and IL-13 level in the lung also significantly decreased. Serum IgE level and lung VCAM-1 expression was also significantly lower in treated animals, but eotaxin protein and mRNA expression in the lung remained unchanged. CONCLUSION: MK exerts its anti-inflammatory effect through the suppression of T helper type-2 (Th2) cytokines. The use of high-dose MK as an anti-inflammatory agent in acute asthma should be further explored.

Acetates↗
PubMed2003

An efficient gene transduction system for studying gene function in primary human dermal fibroblasts and epidermal keratinocytes.

One of the critical challenges for cellular genetic studies in primary human skin cells is lack of a gene delivery system that provides efficient transduction and sustained expression of the transgenes. Due to the limited time of survival in culture, the processes of drug selection and clonal expansion for establishing gene stably expressing cell lines are not a realistic option for primary skin cells. We have examined various gene transduction techniques in primary dermal fibroblasts and epidermal keratinocytes of human skin. We report here that vectors based on the human immunodeficiency virus (HIV, lentivirus) offer more than 90% gene transduction efficiency and sustained expression of transgenes in both human skin cell types. In contrast, most of the commonly used techniques have at best 30% transduction efficiency in these cells. Using two previously reported migration control genes, protein kinase Cdelta and p38alpha-MAPK, as examples, we provide evidence that the unprecedented efficiency of the lentiviral system enables a clear detection of the genes' dominant negative effects, which are otherwise greatly compromised by ordinary transfection techniques. We believe that a wide application of this gene transduction system will greatly benefit studies of gene function in human skin cells.

Fibroblasts↗
PubMed2003

Serum gamma-glutamyltransferase and development of impaired fasting glucose or type 2 diabetes in middle-aged Japanese men.

OBJECTIVE: To investigate the association between serum gamma-glutamyltransferase (GGT) and risk for development of diabetes. DESIGN: Longitudinal study (followed from 1994 to 2001). SETTING: A work site in Japan. SUBJECTS: A total of 2918 Japanese male office workers aged 35-59 years who did not have impaired fasting glucose (IFG) (a fasting plasma glucose concentration of 6.1-6.9 mmol L-1), type 2 diabetes (a fasting plasma glucose concentration of >/=7.0 mmol L-1 or receipt of hypoglycaemic medication), medication for hypertension or hepatitis, alanine aminotransferase concentrations higher than three times the upper limit of the reference range or a history of cardiovascular disease at study entry. MAIN OUTCOME MEASURE: Incidence of IFG or type 2 diabetes over a 7-year period. RESULTS: With adjustment for potential risk factors for diabetes, the relative risk for IFG compared with serum GGT <16 U L-1 was 1.23 (95% CI, 0.79-1.90), 1.50 (CI, 0.97-2.32) and 1.70 (CI, 1.07-2.71) with serum GGT of 16-24, 25-43 and >/=44 U L-1, respectively (P for trend = 0.014). The respective relative risks for type 2 diabetes compared with serum GGT <16 U L-1 were 2.54 (CI, 1.29-5.01), 2.64 (CI, 1.33-5.23) and 3.44 (CI, 1.69-6.70) (P for trend = 0.002). From stratified analyses by body mass index (BMI) and alcohol intake, a stronger linear association between serum GGT and development of IFG or type 2 diabetes was found in men with a BMI >/=23.2 kg m-2 in both those who drank <46 and >/=46 g day-1 of ethanol. CONCLUSIONS: The risk for development of IFG or type 2 diabetes increased in a dose-dependent manner as serum GGT increased in middle-aged Japanese men. The increased relative risk for IFG or type 2 diabetes associated with serum GGT was more pronounced in obese men.

Adult↗
PubMed2003

A rare leucine codon in adpA is implicated in the morphological defect of bldA mutants of Streptomyces coelicolor.

Streptomycetes are mycelial bacteria that produce sporulating aerial hyphae on solid media. Bald (bld) mutants fail to form aerial mycelium under at least some conditions. bldA encodes the only tRNA species able to read the leucine codon UUA efficiently, implying the involvement of a TTA-containing gene in initiating aerial growth. One candidate for such a gene was bldH, because the bldH109 mutant of Streptomyces coelicolor resembles bldA mutants in some aspects. In the work reported here, adpAc, an S. coelicolor gene similar to the Streptomyces griseus A factor-regulated adpAg, was found to complement the bldH109 mutant partially at both single and multiple copies. The sequence of adpAc from the bldH109 mutant revealed a frameshift. A constructed in frame deletion of adpAc conferred a bald colony phenotype, and the mutant behaved like bldA mutants and bldH109 in its pattern of extracellular signal exchange. Both adpAc and adpAg contain a TTA codon. A TTA-free version of adpAc was engineered by replacing the TTA leucine codon with a cognate TTG leucine codon. The adpA(TTATTG) gene could partially restore aerial mycelium formation to a bldA mutant when it was followed in cis by the gene ornA, as in the natural chromosomal arrangement. This indicated that the UUA codon in adpAc mRNA is the principal target through which bldA influences morphological differentiation. It also implied that translational arrest at the UUA codon in adpAc mRNA caused a polar effect on the downstream ornA, and that the poor translation of both genes contributes extensively to the deficiency of aerial mycelium formation in bldA mutants. Unlike the situation in S. griseus, adpAc transcription does not depend on the host's -butyrolactone signalling system, at least in liquid cultures. In addition, sigma factor BldN, which is the homologue of an S. griseus sigma factor AdsA that is absent from adpAg mutants of S. griseus, was present in the constructed adpAc null mutant of S. coelicolor.

Bacterial Proteins↗
PubMed2003

A kinesin mutant with an atypical bipolar spindle undergoes normal mitosis.

Motor proteins have been implicated in various aspects of mitosis, including spindle assembly and chromosome segregation. Here, we show that acentrosomal Arabidopsis cells that are mutant for the kinesin, ATK1, lack microtubule accumulation at the predicted spindle poles during prophase and have reduced spindle bipolarity during prometaphase. Nonetheless, all abnormalities are rectified by anaphase and chromosome segregation appears normal. We conclude that ATK1 is required for normal microtubule accumulation at the spindle poles during prophase and possibly functions in spindle assembly during prometaphase. Because aberrant spindle morphology in these mutants is resolved by anaphase, we postulate that mitotic plant cells contain an error-correcting mechanism. Moreover, ATK1 function seems to be dosage-dependent, because cells containing one wild-type allele take significantly longer to proceed to anaphase as compared with cells containing two wild-type alleles.

Alleles↗
PubMed2003

Entropic interactions on a colloidal sphere near the edge of a terrace.

We calculated the entropic interactions between a colloidal sphere and the hard edge of a terrace by means of Monte Carlo simulation with the acceptance ratio method. Comparison with the Asakura-Oosawa approximation indicates that the AO approximation is more accurate in this model than other known models. Our simulation results are also in qualitative agreement with the experimental results obtained by A.D. Dinsmore et al.

Journal Article↗
PubMed2003

Hypertension and angiogenesis.

Arterial Hypertension (AH) is characterized by reduced nitric oxide (NO) biosynthesis, activation of the Renin-Angiotensin-Aldosteron-System (RAAS), vasoconstriction, and microvascular rarefaction. The latter contributes to target organ damage, especially in left ventricular hypertrophy, and may partially be due to impaired angiogenesis. Angiogenesis, the formation of new microvessels and microvascular networks from existing ones, is a highly regulated process that arises in response to hypoxia and other stimuli and that relieves tissue ischemia. In AH, angiogenesis seems impaired. However, blood pressure alone does not affect angiogenesis, and microvascular rarefaction is present in normotensive persons with a family history for AH. Normal or increased NO in several processes and diseases enables or enhances angiogenesis (e.g. in portal hypertension) and reduced NO biosynthesis (for example, in a rat model of AH, in other disease models in vivo, and in endothelial NO Synthase knock out mice) impairs angiogenesis. Angiogenic growth factors such as Vascular Endothelial Growth Factor (VEGF) and Fibroblast Growth Factor (FGF) induce NO and require NO to elicit an effect. Effector molecules and corresponding receptors of the RAAS either induce (Bradykinin, Angiotensin II) or perhaps inhibit angiogenesis. The pattern of Bradykinin- and Angiotensin II-receptor expression and the capacity to normalize NO biosynthesis may determine whether ACE-inhibitors, Angiotensin II-receptor antagonists and other substances affect angiogenesis. Reconstitution of a normally vascularized tissue by reversal of impaired angiogenesis with drugs such as ACE inhibitors and AT1 receptor antagonists may contribute to successful treatment of hypertension-associated target organ damage, e.g. left ventricular hypertrophy.

Animals↗
PubMed2003

Detection of viable Shiga toxin-producing Escherichia coli by quantitative competitive polymerase chain reaction.

With the use of Escherichia coli O157:H7 as a model, a procedure for the quantitative detection of viable Shiga toxin-producing E. coli (STEC) in broth and cooked ground beef enrichments with multiple-time point quantitative competitive polymerase chain reaction (QC-PCR) was developed. The A subunit (a 401-bp fragment) of the stx2 gene was chosen as a target sequence. Immunomagnetic separation (IMS) was used to isolate and concentrate cells from ground beef enrichments. Cell viability was confirmed on the basis of the quantitative increase in the signal of target bands from QC-PCR across multiple time points. The application of IMS increased detection limits relative to those for QC-PCR without IMS. E. coli O157:H7 inoculated at 0.20 CFU/g of cooked ground beef (25 g of ground beef plus 225 ml of Bacto modified EC medium plus novobiocin) was detected and confirmed to be viable in <15 h. A DNA-based molecular approach can be used to determine cell viability.

Colony Count, Microbial↗
PubMed2002

Study of Nd3+, Pd2+, Pt4+, and Fe3+ dopant effect on photoreactivity of TiO2 nanoparticles.

The metallorganic chemical vapor deposition method was successfully used to synthesize pure TiO(2) and Nd(3+)-, Pd(2+)-, Pt(4+)-, and Fe(3+)-doped TiO(2) nanoparticles. Polycrystalline TiO(2) structure was verified with x-ray diffraction, which showed typical characteristic anatase reflections without any separate dopant-related peaks. Transmission electron microscopy observations confirmed the existence of homogeneously distributed 22 +/- 3 nm TiO(2) nanoparticles. The particle size remained the same for the doped samples. The doping level of transition metals was kept at approximately 1 atomic percent, which was determined by x-ray photoelectron spectra and energy dispersive x-ray spectroscopy. The effects of different types of dopants on the photocatalytic activity were revealed by the degradation of 2-chlorophenols with an UV light source. The photocatalytic efficiency was remarkably enhanced by the introduction of Pd(2+) and Nd(3+). Nd(3+)-doped TiO(2) showed the largest enhancement. However, Pt(4+) changed the 2-chlorophenol degradation rate only slightly, and Fe(3+) was detrimental to this process. These effects were related to the position of the dopants in the nanoparticles and the difference in their ionic radii with respect to that of Ti(4+).

Journal Article↗
PubMed2002

T1 efficacy of EVP-ABD: a potential manganese-based MR contrast agent for hepatic vascular and tissue phase imaging.

PURPOSE: To evaluate the T1 efficacy of EVP-ABD, a new manganese (Mn)-based contrast agent, for vascular and liver tissue enhancement in comparison with currently approved agents. MATERIALS AND METHODS: Ten Yorkshire pigs (body weight, 26 -46 kg) were used for the efficacy evaluation, nine for kinetic T1 evaluation (three each agent) and one for post EVP-ABD imaging. With a fast imaging scheme to monitor T1 values of blood and liver, 10 micromol/kg EVP-ABD was injected intravenously and compared with gadopentetate dimeglumine (Magnevist, GdDTPA) and mangafodipir trisodium (Teslascan, mangafodipir trisodium) at routine clinical dosages. All were imaged with 3D T1 Gradient Recalled Echo (GRE) sequence (TR/TE/alpha = 3.8/1.6/25 degrees ) prior to and 10 minutes post injection using a 1.5-T whole-body scanner. Additional high-resolution 2D liver images (TR/TE/alpha = 50/4.6/40 degrees ) and arterial phase images of the upper aorta were acquired from the pig for post EVP-ABD imaging. RESULTS: At 10 micromol/kg, EVP-ABD provided a dramatic decline in blood T1, comparable to 0.1 mmol/kg GdDTPA, followed by a rapid return to blood baseline T1 values. In addition to the blood enhancement phase, EVP-ABD achieved a 70% reduction in liver T1 within 2 minutes postadministration, with an imaging window of at least 2 hours. A substantially improved signal-to-noise ratio (SNR) was observed in both the 2D and 3D liver images postcontrast. CONCLUSION: EVP-ABD demonstrated peak vascular enhancement similar to GdDTPA and prolonged specific liver enhancement exceeding mangafodipir trisodium. EVP-ABD has favorable T1 enhancing characteristics with the potential to allow for a comprehensive liver evaluation.

Animals↗
PubMed2002

Pyrolysis-gas chromatography/mass spectrometry of fractions separated from a low-temperature coal tar: an attempt to develop a general method for characterising structures and compositions of heavy hydrocarbon liquids.

A low-temperature coal tar has been fractionated by column chromatography into acetonitrile, pyridine and 1-methyl-2-pyrrolidinone- (NMP) solubles. The tar and its fractions have been examined by pyrolysis-gas chromatography/mass spectrometry (GC/MS). Fractionation by planar chromatography was also carried out for purposes of comparison. Molecular masses of the fractions were estimated by size-exclusion chromatography (SEC), and bulk structural characterisation was carried out by (13)C-NMR and UV-fluorescence spectrometry. SEC showed that the fractions shifted to progressively shorter elution times (higher apparent masses) with diminishing solubility, i.e. from acetonitrile to NMP solubles. UV-fluorescence spectra showed parallel shifts to longer wavelengths and lower fluorescence quantum yields, indicating increasing sizes of aromatic ring systems and increasingly complex molecules. GC/MS analysis of the tar showed alkanes from C10 to C32 and extensive series of alkylated aromatics, phenols, indenes, naphthalenes, phenanthrenes and fluoranthenes. Pyrolysis-GC/MS results for the acetonitrile solubles closely resembled the data for the tar sample, with extensive series of alkylated benzenes, phenols and naphthalenes as well as alkanes from C16 to C28. The pyridine-soluble fraction showed no significant aromatic pyrolysis products and only relatively weak signals for alkanes between C16 and C27. The NMP-soluble fraction showed even less overall signal, with no significant aromatic components and weak signals for alkanes between C21 and C25, even though (13)C-NMR analyses showed that approximately half of the carbon detected was aromatic. The aliphatics are assumed to provide bridging structures between polycyclic aromatic (PCA) ring systems.

Journal Article↗