Biomedical subjects

Y Liu

Publications and source records attributed to Y Liu.

At least 775 records · Page 43Linked to original sources

PubMed2000

Human papillomavirus type 16 E6 and E7 proteins inhibit differentiation-dependent expression of transforming growth factor-beta2 in cervical keratinocytes.

Infection with high-risk human papillomaviruses (HPVs) represents a major risk factor for the development of cervical cancer. The HPV-16 E6 and E7 proteins are highly expressed in differentiating keratinocytes, where they inactivate the p53 and retinoblastoma (pRb) proteins, two important transcriptional regulators. We have used cDNA expression arrays to identify global alterations in gene expression induced by E6 and E7 in differentiating cultures of human cervical keratinocytes. We show that E6 and E7 decrease expression of TGF-beta2 mRNA and alter expression of multiple TGF-beta-responsive genes involved in cell cycle regulation, apoptosis, and tissue remodeling. E6 and E7 inhibited expression of TGF-beta2 RNA 7-fold (relative effectiveness, E6/ E7 > E6 > E7 > control) and decreased secretion of biologically active TGF-beta2 by 70-80% (reduced from 70 to 10 pg/10(6) cells/24 h). Downregulation occurred through p53- and pRb-dependent pathways. In contrast, E6 and E7 did not alter expression of TGF-beta1 and TGF-beta3. Down-regulation of TGF-beta2 was biologically relevant because the addition of recombinant cytokine (10-200 pg/ml) to E6/E7-expressing cells restored expression of TGF-P-responsive genes, inhibited growth of keratinocytes, and decreased immortalization by E6 and E7. These results suggest that TGF-32- and TGF-3-responsive genes are important targets for the HPV-16 E6 and E7 oncoproteins in differentiating cervical keratinocytes.

Blotting, Northern↗

PubMed2000

Cloning and functional expression of two families of beta-subunits of the large conductance calcium-activated K+ channel.

We report here a characterization of two families of calcium-activated K(+) channel beta-subunits, beta2 and beta3, which are encoded by distinct genes that map to 3q26.2-27. A single beta2 family member and four alternatively spliced variants of beta3 were investigated. These subunits have predicted molecular masses of 27. 1-31.6 kDa, share approximately 30-44% amino acid identity with beta1, and exhibit distinct but overlapping expression patterns. Coexpression of the beta2 or beta3a-c subunits with a BK alpha-subunit altered the functional properties of the current expressed by the alpha-subunit alone. The beta2 subunit rapidly and completely inactivated the current and shifted the voltage dependence for activation to more polarized membrane potentials. In contrast, coexpression of the beta3a-c subunits resulted in only partial inactivation of the current, and the beta3b subunit conferred an apparent inward rectification. Furthermore, unlike the beta1 and beta2 subunits, none of the beta3 subunits increased channel sensitivity to calcium or voltage. The tissue-specific expression of these beta-subunits may allow for the assembly of a large number of distinct BK channels in vivo, contributing to the functional diversity of native BK currents.

Alternative Splicing↗

PubMed2000

m5C RNA and m5C DNA methyl transferases use different cysteine residues as catalysts.

A family of RNA m(5)C methyl transferases (MTases) containing over 55 members in eight subfamilies has been identified recently by an iterative search of the genomic sequence databases by using the known 16S rRNA m(5)C 967 MTase, Fmu, as an initial probe. The RNA m(5)C MTase family contained sequence motifs that were highly homologous to motifs in the DNA m(5)C MTases, including the ProCys sequence that contains the essential Cys catalyst of the functionally similar DNA-modifying enzymes; it was reasonable to assign the Cys nucleophile to be that in the conserved ProCys. The family also contained an additional conserved Cys residue that aligns with the nucleophilic catalyst in m(5)U54 tRNA MTase. Surprisingly, the mutant of the putative Cys catalyst in the ProCys sequence was active and formed a covalent complex with 5-fluorocytosine-containing RNA, whereas the mutant at the other conserved Cys was inactive and unable to form the complex. Thus, notwithstanding the highly homologous sequences and similar functions, the RNA m(5)C MTase uses a different Cys as a catalytic nucleophile than the DNA m(5)C MTases. The catalytic Cys seems to be determined, not by the target base that is modified, but by whether the substrate is DNA or RNA. The function of the conserved ProCys sequence in the RNA m(5)C MTases remains unknown.

Amino Acid Sequence↗

PubMed2000

Reading Chinese characters for meaning: the role of phonological information.

Two experiments with the Stroop paradigm were conducted to investigate the role of phonological codes in access to the meaning of Chinese characters. Subjects named the ink color of viewed characters or color patches. Key items were color characters, their homophones with the same tone, homophones with different tones, and semantic associates. Apart from finding the usual Stroop interference effect, homophones produced significant interference in the incongruent condition, provided that they had the same tone as the color characters. The interference effect from homophones, however, was significantly smaller than that from color characters. Semantic associates generated an interference effect in the incongruent condition, an effect of the same magnitude as the effect from the same-tone homophones. Finally, in the congruent conditions, all the key items yielded facilitations compared to neutral controls, though the facilitation from color characters was larger than the facilitations from other types of characters. These findings suggest that phonological codes in Chinese are activated obligatorily and provide early sources of constraint in access to meaning.

Cognition↗

PubMed2000

Differential gene expression in nasopharyngeal carcinoma cells.

Nasopharyngeal carcinoma (NPC) is a common cancer among southern Chinese. The profile of gene expression in NPC cells is largely unknown. In this study, we have examined differential gene expression in non-malignant and malignant nasopharyngeal epithelial (NPE) cells using a cDNA array hybridization method. A total of 42 genes were identified to be expressed in either non-malignant and malignant NPE cells or both. Thirteen of these genes were overexpressed in malignant NPE cells. These includes nuclear factor (NF90), FOS-related antigen 1 (FRA- 1), cytoplasmic dynein light chain (HDLC1), replication factor C (RFC1), nucleoside diphosphate kinase B, UV excision repair protein (RAD23A), insulin-like growth factor receptor II, transcription initiation factor TFIID subunit (TAFII31), growth factor receptor-bound protein 2 (GRB2), UV excision repair protein (RAD23B), glutathione peroxidase, Y box binding protein 1 and heat shock protein 86. In contrast, expression of nine genes was suppressed in malignant NPE cells. These includes calgranulin A, calgranulin B, neutrophil activating protein (ENA-78), heat shock protein 27, integrin beta-1, integrin beta-4, cyclin-dependent kinase inhibitor 1A (p21), interleukin-8 and tyrosine protein kinase receptor (RET). Differential expression of calgranulin A, calgraunlin B, ENA-78, FRA-1 and NF90 in non-malignant and malignant nasopharyngeal epithelial cells was confirmed by RT-PCR analysis.

Calcium-Binding Proteins↗

PubMed2000

Applications of a sulfonated-polymer wall-modified open-tubular fused-silica capillary in capillary zone electrophoretic separations.

A fused-silica capillary that is wall-modified via chemically bonding a sulfonated polymer to the capillary wall has a uniform negative charge density on its surface and produces an electroosmotic flow (EOF) greater than 4 x 10(-4) cm2 V(-1) s(-1) The EOF is nearly independent of buffer pH over the pH range of 2 to 10 and is lower than the EOF obtained for the bare fused-silica capillary at the more basic pH but is higher at the more acidic buffer pH. Optimization of buffer pH can be based on analyte pKa values to improve the overall quality of the capillary zone electrophoresis (CZE) separation of complex mixtures of weak acid and base analytes. Because of the high EOF in an acidic buffer, the capillary is useful for the separation of weak organic bases which are in their cation forms in the acidic buffer. EOF for the sulfonic acid bonded phase capillary can be adjusted via buffer additives such as organic solvent, tetraalkylammonium salts, multivalent cations and alkylsulfonic acids. The advantages of utilizing buffer pH and the EOF buffer modifiers to enhance migration time, selectivity, and resolution in CZE separations with this capillary are illustrated using a series of test analyte mixtures of inorganic anions, carboxylic acids, alkylsulfonic acids, benzenesulfonic acids, sulfas, pyridines, anilines or small-chain peptides.

Alkanesulfonic Acids↗

PubMed2000

Analysis of oligomannuronic acids and oligoguluronic acids by high-performance anion-exchange chromatography and electrospray ionization mass spectrometry.

Oligomannuronic acids and oligoguluronic acids were prepared by enzymatic hydrolysis of alginate with alginate lyases. The oligosaccharides generated up to degree of polymerization 16 were characterized by high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) and electrospray ionization mass spectrometry (ESI-MS). Acetate buffer linear gradients were used as mobile phases for separations of oligosaccharides. ESI-MS and HPAEC-PAD are very effective for the analysis and characterization of anionic oligosaccharides.

Anion Exchange Resins↗

PubMed2000

Chronic combined exposure to cadmium and arsenic exacerbates nephrotoxicity, particularly in metallothionein-I/II null mice.

Cadmium (Cd) and arsenic (As) are important inorganic toxicants in the environment. Humans certainly have the potential to be exposed to the mixtures of Cd and As, but the toxicological interactions of these inorganic mixtures are poorly defined. Metallothionein (MT) is a cysteine-rich, metal-binding protein that plays an important role in Cd detoxication, but its role in As toxicity is less certain. To examine the role of MT in Cd- and/or As-induced nephrotoxicity, MT-I/II-knockout (MT-null) mice and background-matched wild-type (WT) mice were fed CdCl(2) (100 ppm Cd) in the diet, NaAsO(2) (22.5 ppm As) in the drinking water, or Cd plus As for 4 months. Subsequently, nephrotoxicity was examined by morphological and biochemical techniques. Chronic exposure to Cd produced more renal toxicity than As, and the combination of Cd and As produced even more renal injury than caused by either of the chemicals given alone. In mice receiving Cd plus As, proximal tubule degeneration and atrophy, glomerular swelling and interstitial fibrosis were more severe than those produced by either inorganic. Furthermore, lack of MT rendered MT-null mice more sensitive than WT mice to the nephrotoxicity produced by chronic Cd- and/or As-exposure. MT-null mice were especially susceptible to the toxicity produced by the combination of Cd and As, as evidenced by decreased body weight, enzymuria, glucosuria, proteinuria and nephropathy. In conclusion, this study indicates that As may potentiate Cd nephrotoxicity during the long-term, combined exposure, and that intracellular MT plays a role in decreasing the nephropathy of combined exposure to Cd and As.

Acetylglucosaminidase↗

PubMed2000

Novel method for the labeling of distant neuromuscular junctions.

Essential to understanding the roles proteins and structural elements play at the synapse is to understand the development, remodeling and reinnervation of peripheral neuromuscular junctions. It has, however, been a challenging task to label and visualize neuromuscular junctions. In this paper we demonstrate how adenovirus technology can be combined with intraspinal microinjection techniques to follow both the development and the reinnervation of a distant peripheral neuromuscular junction in the rat. A recombinant adenovirus containing VAMP-2 (synaptobrevin-2) was fused to the green fluorescent protein (GFP) and microinjected into the region of the lumbar motor neurons. We were able to follow the neuronal incorporation, axonal transport and synaptic localization of the GFP-VAMP-2 using fluorescence microscopy. GFP-VAMP-2 was found in neuronal cell bodies, selected sciatic nerve axons and was concentrated in the presynaptic nerve terminal. During reinnervation of the neuromuscular junction, GFP-VAMP-2 allows us to follow the time course of junctional reinnervation. Thus, the microinjection of microliter amounts of labeled recombinant virus into locations far distant from target regions can be used to efficiently study the formation of neuromuscular junctions with a minimum of trauma to the animal.

Adenoviridae↗

PubMed2000

Adsorption of PBTCA on Alumina Surfaces and Its Influence on the Fractal Characteristics of Sediments.

The adsorption of 2-phosphonobutane-1,2,4-tricarboxylic acid (PBTCA) on Al(2)O(3) powder has been studied as a function of pH and concentration. The adsorption density of PBTCA is found to decrease with an increase of pH. Zeta potential measurements show that the addition of PBTCA results in a dramatic increase in the absolute zeta potential, as well as a shift of the isoelectric point to the more acidic region. PBTCA considerably enhances the stability of the alumina suspension via an electrostatic mechanism. The surface properties of alumina suspensions are examined by using Auger electron microscopy and FTIR analysis. Chemical interactions take place at the solid/water interface by forming complexes between Al(3+) ions and PBTCA. The dispersing ability of PBTCA is believed to be related to its high adsorption ability and the high number of dissociable protons. The morphology of the sediments is observed with scanning electron microscopy. It is found that the sediment surfaces exhibit fractal characteristics. The fractal dimension values of sediments are correlated with PBTCA concentration in the experimental range. Copyright 2000 Academic Press.

Journal Article↗

PubMed2000

Metabotropic glutamate receptor subtypes involved in cardiovascular regulation in the rostral ventrolateral medulla of rats.

We have previously reported that metabotropic glutamate receptors (mGluR) participate in cardiovascular regulation in the rostral ventrolateral medulla (RVLM) of rats. In the present study, we have tried to elucidate which subtype of mGluR contributes to cardiovascular responses elicited by L-glutamate in the RVLM. Adult male Wistar rats (348 +/- 11 g, n = 21) were anesthetized and artificially ventilated. Microinjections of agonists and antagonists for each mGluR subtype were done into unilateral RVLM. Each of group I, II and III mGluR agonist (1 nmol/50 nl) produced significant increases in arterial pressure (18 +/- 2, 9 +/- 2 and 34 +/- 3 mmHg, respectively) and heart rate (18 +/- 4, 13 +/- 3 and 33 +/- 12 bpm, respectively). Microinjections of group I, II and III mGluR antagonists failed to inhibit the cardiovascular responses induced by subsequently injected agonists. However, group I antagonist [(RS)-1-aminoindan-1,5-dicarboxylic acid] elicited transient increases in arterial pressure and heart rate, followed by decreases in both variables (-19 +/- 4 mmHg and -22 +/- 4 bpm). These results suggest that all of three subtypes of mGluR participate in cardiovascular responses induced by L-glutamate, and group I mGluR may play an important role in the maintenance of arterial pressure.

Animals↗

PubMed2000

Measurement of isotopes of light rare earth elements in the form of oxide ions: a new development in thermal ionization mass spectrometry

An isobaric interference free thermal ionization mass spectrometry (TIMS) method is presented here for the precise isotope measurements of light rare earth elements (LREEs). LREE isotopes can be measured in the form of monoxide ion (MO+), but the previous determinations of LREE isotopes in the form of oxide were hindered, notably by the 17O and 18O related isobaric interferences. In the present study, 16O-enriched oxygen was introduced into the ion source chamber via a specially designed gas inlet system. It was observed that the isobaric interferences caused by 17O and 18O were remarkably reduced by up to 6 and 70 times, respectively, compared to those caused by natural oxygen bleeding. Isotopic ratios of La, Ce, and Nd obtained without isobaric interference corrections show the excellent precision and accuracy in the present study. The results also shed some light on the formation mechanism of the measured oxide species in TIMS. Furthermore it promises more precise determination of LREE abundances and La-Ce and Sm-Nd ages in geochemistry and cosmochemistry.

Journal Article↗

PubMed2000

The temporal response of the brain after eating revealed by functional MRI.

After eating, the human brain senses a biochemical change and then signals satiation, but precisely when this occurs is unknown. Even for well-established physiological systems like glucose-insulin regulation, the timing of interaction between hormonal processes and neural events is inferred mostly from blood sampling. Recently, neuroimaging studies have provided in vivo information about the neuroanatomical correlates of the regulation of energy intake. Temporal orchestration of such systems, however, is crucial to the integration of neuronal and hormonal signals that control eating behaviour. The challenge of this functional magnetic resonance imaging study is to map not only where but also when the brain will respond after food ingestion. Here we use a temporal clustering analysis technique to demonstrate that eating-related neural activity peaks at two different times with distinct localization. Importantly, the differentiated responses are interacting with an internal signal, the plasma insulin. These results support the concept of temporal parcellation of brain activity, which reflects the different natures of stimuli and responses. Moreover, this study provides a neuroimaging basis for detecting dynamic processes without prior knowledge of their timing, such as the acute effects of medication and nutrition in the brain.

Adult↗

PubMed2000

Inhibitors of COPI and COPII do not block PEX3-mediated peroxisome synthesis.

In humans, defects in peroxisome biogenesis are the cause of lethal diseases typified by Zellweger syndrome. Here, we show that inactivating mutations in human PEX3 cause Zellweger syndrome, abrogate peroxisome membrane synthesis, and result in reduced abundance of peroxisomal membrane proteins (PMPs) and/or mislocalization of PMPs to the mitochondria. Previous studies have suggested that PEX3 may traffic through the ER en route to the peroxisome, that the COPI inhibitor, brefeldin A, leads to accumulation of PEX3 in the ER, and that PEX3 overexpression alters the morphology of the ER. However, we were unable to detect PEX3 in the ER at early times after expression. Furthermore, we find that inhibition of COPI function by brefeldin A has no effect on trafficking of PEX3 to peroxisomes and does not inhibit PEX3-mediated peroxisome biogenesis. We also find that inhibition of COPII-dependent membrane traffic by a dominant negative SAR1 mutant fails to block PEX3 transport to peroxisomes and PEX3-mediated peroxisome synthesis. Based on these results, we propose that PEX3 targeting to peroxisomes and PEX3-mediated peroxisome membrane synthesis may occur independently of COPI- and COPII-dependent membrane traffic.

ATP-Binding Cassette Transporters↗

PubMed2000

Analysis of K-ras and p53 mutations in mesotheliomas from humans and rats exposed to asbestos.

Malignant mesothelioma is known to be associated with asbestos exposure. However, the mechanism of mesothelial carcinogenesis in relation to the activation of proto-oncogenes or inactivation of tumor suppressor genes remains unclear. In this study, the PCR-Primer Introduced Restriction Site (PCR-PIRS) assay was employed to examine mutations in the K-ras proto-oncogene in mesothelioma tissues from workers exposed to asbestos and from rats treated with asbestos. Mutations in exons 5-8 of the p53 tumor suppressor gene were determined by direct DNA sequence analysis. Results of the PCR-PIRS analysis revealed no mutations in codons 12, 13 or 61 of the K-ras gene in any of the 17 human or 22 rat mesothelioma tissue samples. These results were confirmed by direct DNA sequence analysis. No mutations were found in exons 5-8 of the p53 gene in any of the mesothelioma tissue samples analyzed. These results and the results reported by others indicate that the K-ras proto-oncogene and p53 tumor suppressor gene may not play a critical role in the induction of mesothelioma by asbestos either in humans or in rats.

Animals↗

PubMed2000

Transforming growth factor-beta 1 suppresses serum deprivation-induced death of A549 cells through differential effects on c-Jun and JNK activities.

Transforming growth factor (TGF)-beta1, a pleiotropic cytokine involved in regulating growth and differentiation, can exert both pro-apoptotic and anti-apoptotic effects depending on the cell type or circumstances. We observed that TGF-beta1 blocked apoptosis resulting from serum withdrawal in A549 human lung carcinoma cells. This was associated with suppression of JNK activation that occurs concomitant with the onset of apoptosis in the absence of TGF-beta1, suggesting that JNK plays an active role in the death process and that TGF-beta1 exerts its protective influence by altering JNK activity. Overexpression of a dominant negative mutant form of SEK1, an upstream activator of JNK, likewise suppressed JNK activation and inhibited apoptosis. Investigation of early events following TGF-beta1 treatment revealed an early induction and phosphorylation of c-Jun that was absent in cells subjected to serum withdrawal alone. That TGF-beta1-induced expression of c-Jun is important for survival was supported by the finding that overexpression of non-phosphosphorylatable dominant negative mutant c-Jun, c-Jun(S73A), attenuated the protective influence of TGF-beta1. Our findings suggest that JNK activation is a late but essential event in serum deprivation-induced apoptosis in A549 cells. TGF-beta1 prevents apoptosis, in part, through the early induction and phosphorylation of c-Jun, which in turn results in attenuated JNK activation.

Alanine↗

PubMed2000

Inhibition of flap endonuclease 1 by flap secondary structure and relevance to repeat sequence expansion.

Recent genetic evidence indicates that null mutants of the 5'-flap endonuclease (FEN1) result in an expansion of repetitive sequences. The substrate for FEN1 is a flap formed by natural 5'-end displacement of the short intermediates of lagging strand replication. FEN1 binds the 5'-end of the flap, tracks to the point of annealing at the base of the flap, and then cleaves. Here we examine mechanisms by which foldback structures within the flap could contribute to repeat expansions. Cleavage by FEN1 was reduced with increased length of the foldback. However, even the longest foldbacks were cleaved at a low rate. Substrates containing the repetitive sequence CTG also were cleaved at a reduced rate. Bubble substrates, likely intermediates in repeat expansions, were inhibitory. Neither replication protein A nor proliferating cell nuclear antigen were able to assist in the removal of secondary structure within a flap. We propose that FEN1 cleaves natural foldbacks at a reduced rate. However, although the cleavage delay is not likely to influence the overall process of chromosomal replication, specific foldbacks could inhibit cleavage sufficiently to result in duplication of the foldback sequence.

Base Sequence↗

PubMed2000

Purification of human prothrombin from Nitschmann fraction III using DEAE membrane radial flow chromatography.

A DEAE membrane radial column (4x5 cm) was used to purify human prothrombin from Nitschmann fraction III and performances of the column were investigated. Sample flow-rate of 20-30 ml/min (i.e., 24-36 bed volumes/h) and elution velocity up to 40 ml/min (i.e., 48 bed volumes/h) were obtained without compromising the separation efficiency. Its breakthrough capacity was one-third to half that of the commercial DEAE Sepharose Fast Flow media. It was concluded that the novel column is an attractive alternate to traditional axial column.

Chromatography, Liquid↗