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Y Cai

Publications and source records attributed to Y Cai.

At least 181 records · Page 10Linked to original sources
PubMed1998

Somatic deletion of the 5' ends of both the COL4A5 and COL4A6 genes in a sporadic leiomyoma of the esophagus.

Leiomyomata of the esophagus are sporadic benign tumors of unknown etiology. We studied a collection of nine tumors for the expression of extracellular matrix components and found the same aberrant expression pattern as previously observed in inherited diffuse leiomyomatosis. We demonstrate here the occurrence of a somatic deletion at the COL4A5/COL4A6 locus at Xq22 in a frozen leiomyoma sample. These data confirm the hypothesis that the same underlying etiology is responsible for circumscribed smooth muscle proliferation in sporadic leiomyomata as for diffuse smooth muscle cell proliferation in inherited diffuse leiomyomatosis.

Basement Membrane↗
PubMed1997

Molecular cloning, cDNA sequence analysis, and chromosomal localization of mouse Pkd2.

The gene responsible for the second form of autosomal dominant polycystic kidney disease, PKD2, has recently been identified. We now describe the cloning, genomic localization, cDNA sequence, and expression analysis of its murine homologue, Pkd2. The cloned cDNA sequence is 5134 bp long and is predicted to encode a 966-amino-acid integral membrane protein with six membrane-spanning domains and intracellular NH2 and COOH termini. Pkd2 is highly conserved with 91% identity and 98% similarity to polycystin-2 at the amino acid level. Pkd2 mRNA is widely expressed in mouse tissues. Pkd2 maps to mouse Chromosome 5 and is excluded as a candidate gene for previously mapped mouse mutations resulting in a polycystic kidney phenotype.

Amino Acid Sequence↗
PubMed1997

Characterization of the exon structure of the polycystic kidney disease 2 gene (PKD2).

PKD2, the gene defective in the second form of autosomal dominant polycystic kidney disease (ADPKD), has been identified by positional cloning and found to encode an integral membrane protein with similarity to the gene for the more common form of ADPKD and to calcium channels. We have determined the exon-intron structure of the PKD2 gene. PKD2 is encoded in at least 15 exons with the translation start site in exon 1. All the splice acceptor and donor sites conform to the AG/GT rule. We have designed a series of intronic oligonucleotide primers for amplifying the entire coding sequence from genomic DNA in segments well suited to mutation analysis using conventional screening strategies such as SSCA or heteroduplex analysis.

Cloning, Molecular↗
PubMed1997

Inscuteable and Staufen mediate asymmetric localization and segregation of prospero RNA during Drosophila neuroblast cell divisions.

When neuroblasts divide, inscuteable acts to coordinate protein localization and mitotic spindle orientation, ensuring that asymmetrically localized determinants like Prospero partition into one progeny. staufen encodes a dsRNA-binding protein implicated in mRNA transport in oocytes. We demonstrate that prospero RNA is also asymmetrically localized and partitioned during neuroblast cell divisions, a process requiring both inscuteable and staufen. Inscuteable and Staufen interact and colocalize with prospero RNA on the apical cortex of interphase neuroblasts. Staufen binds prospero RNA in its 3'UTR. Our findings suggest that Inscuteable nucleates an apical complex and is required for protein localization, spindle orientation, and RNA localization. Stau, as one component of this complex, is required only for RNA localization. Hence staufen also acts zygotically, downstream of inscuteable, to effect aspects of neuroblast asymmetry.

Animals↗
PubMed1997

The high affinity alphaIIb beta3 integrin is involved in invasion of human melanoma cells.

Integrins play an important role in mediating tumor cell-extracellular matrix (ECM) and tumor cell-endothelial cell interactions. The integrin alphaIIb beta3 (GPIIb-IIIa) is expressed on the surface of platelets in an inactive state and requires a conformational change to recognize extracellular matrix proteins such as fibrinogen, fibronectin, vitronectin, and others. In this study, we questioned whether human melanoma cells express the alphaIIb beta3 integrin. Reverse transcription-PCR/Southern blotting, Northern blotting, and dot blotting demonstrated the presence of the platelet-type alphaIIb beta3 integrin in human melanoma WM 983B, WM 983A, and WM 35 cells. AP-2, a monoclonal antibody (mAb) to alphaIIb beta3, positively stained two human melanoma specimens, indicating expression of this integrin in vivo. Phorbol 12-myristate 13-acetate and 12(S)-hydroxyeicosatetraenoic acid, two activators of protein kinase C, stimulated adhesion of melanoma cells to immobilized fibronectin and PAC-1, a mAb to alphaIIb beta3. PAC-1 specifically recognizes the conformationally active form of platelet alphaIIb beta3. Phorbol 12-myristate 13-acetate-stimulated adhesion of WM 983B cells to PAC-1 was completely blocked by an RGD peptide, thus providing evidence that tumor cell adhesion to PAC-1 is mediated via the alphaIIb beta3 integrin but not the Fc receptor. Confocal immunofluorescent studies demonstrated that fibronectin-adherent melanoma cells possess an intracellularly localized pool of high-affinity alphaIIb beta3. Invasion of WM 983B cells through fibronectin was stimulated by 12(S)-hydroxyeicosatetraenoic acid, and this stimulated invasion was blocked by the mAb PAC-1. The data suggest that melanoma cells express the high-affinity alphaIIb beta3 integrin, which is involved in tumor invasion.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1997

A simple program for simulating the responses of neurons with arbitrarily structured and active dendritic trees.

We describe a simple program to simulate neural responses. This program is based on the compartmental approach, in which all compartments of a neuron (i.e. axon, soma or dendrite) are represented by the same basic electrical structure. A parameter file is used to store the model parameters, including the nonlinear channel characteristics of each compartment. The model is then automatically configured according to the values specified in the parameter file. The computation of the conductance of each active channel over time is handled by a unique subroutine optimized according to the kinetics of each channel. The equations for arbitrarily structured trees are solved implicitly using a simple algorithm similar to that of Hines (Hines, M. (1984) Int. J. Biomed. Comput., 15:69-76). The output of the model uses PostScript format. The advantage of this program is that it is small in size, simple to use, efficient, and is platform independent.

Algorithms↗
PubMed1997

Plant polyphenols: biologically active compounds or non-selective binders to protein?

Twenty phenolic compounds, representatives of proanthocyanidins and gallic acid/hexahydroxyldiphenic acid esters of glucose, have been assessed for their ability to inhibit binding of specific radioligands to 16 receptors. The receptors utilized were alpha 1-, alpha 2- and beta-adrenoceptors, adenosine 1, dopamine 1 and 2, muscarinic, Ca2+ channel, sulphonylureas, 5HT1, 5HT1A, 5HT2, histamine 1, benzodiazepine, opiate and Na+/K/ATPase. These phenolic compounds failed to inhibit ligands binding to 10 of the receptors under the test conditions. The most susceptible receptors to phenolic binding were beta-adrenergic, 5HT1 and opiate. Some of the compounds tested showed selectivity for a single or for two receptors. The inhibition of binding of radioligands to receptors by the phenolic compounds cannot be explained solely in terms of phenolic-protein binding. The results indicate that the removal of tannins from plant extracts prior to screening for receptor activities may result in missing biologically active compounds with specificity of action.

Flavonoids↗
PubMed1997

PKD1 interacts with PKD2 through a probable coiled-coil domain.

Autosomal dominant polycystic kidney disease (ADPKD) describes a group of at least three genetically distinct disorders with almost identical clinical features that collectively affects 1:1,000 of the population. Affected individuals typically develop large cystic kidneys and approximately one half develop end-stage renal disease by their seventh decade. It has been suggested that the diseases result from defects in interactive factors involved in a common pathway. The recent discovery of the genes for the two most common forms of ADPKD has provided an opportunity to test this hypothesis. We describe a previously unrecognized coiled-coil domain within the C terminus of the PKD1 gene product, polycystin, and demonstrate that it binds specifically to the C terminus of PKD2. Homotypic interactions involving the C terminus of each are also demonstrated. We show that naturally occurring pathogenic mutations of PKD1 and PKD2 disrupt their associations. We have characterized the structural basis of their heterotypic interactions by deletional and site-specific mutagenesis. Our data suggest that PKD1 and PKD2 associate physically in vivo and may be partners of a common signalling cascade involved in tubular morphogenesis.

Humans↗
PubMed1997

Autoantibody reactive with the human general transcription factor TFIIF in sera from patients with autoimmune disorders.

Transcription factor (TF) IIF, a heteromeric protein composed of two subunits, RAP30 and RAP74, is required for both specific initiation and elongation of mRNA synthesis by RNA polymerase II. We have identified high titre of specific autoantibodies against the RAP74 subunit of TFIIF in sera from patients with systemic lupus erythematosus (SLE) as well as those with rheumatoid arthritis (RA), periarteritis nodosa (PN), Sjögren's syndrome (SS), dermatomyositis (DM), and mixed connective tissue disease (MCTD) by Western blot or immunoprecipitation. The epitopes recognized by autoantibodies were shown to be preferentially clustered at the central charged region. Anti-RAP74 autoantibody was shown to suppress the activity of TFIIF-stimulated elongation of mRNA synthesis by RNA polymerase II. It is concluded that some patients with autoimmune disorders develop specific autoantibodies against the RAP74 subunit of TFIIF.

Adult↗
PubMed1997

Effect of NaCl-tolerant lactic acid bacteria and NaCl on the fermentation characteristics and aerobic stability of silage.

NaCl-tolerant lactic acid bacteria (LAB) strains LC-10 (Lactobacillus casei) and LP-15 (Lact. plantarum) and NaCl were used as additives to sorghun (Sorghum bicolor). Numbers of LAB were significantly (P < 0.05) higher in all the additive-treated silages than in the control silage at an early stage of ensiling. During the fermentation process, addition of NaCl or LAB effectively inhibited the growth of aerobic bacteria and clostridia, but not yeasts. All the additive-treated silages had significantly (P < 0.05) lower pH, ammonia nitrogen content, dry matter loss and gas production but significantly (P < 0.05) higher lactic acid content and residual water soluble carbohydrates compared with the control silage. The improvement in silage quality was in the order: LAB > NaCl > control. Yeast counts were high in all additive-based silages and they increased during the exposure of the silages to air. As a result, these silages suffered aerobic deterioration, whereas the control silage was stable. The results confirmed that the NaCl or LAB improved fermentation quality but did not prevent aerobic deterioration of the silage.

Aerobiosis↗
PubMed1997

Isolation and characterization of nisin-producing Lactococcus lactis subsp. lactis from bean-sprouts.

Bacterial isolates from bean-sprouts were screened for anti-Listeria monocytogenes bacteriocins using a well diffusion method. Thirty-four of 72 isolates inhibited the growth of L. monocytogenes Scott A. One, HPB 1688, which had the biggest inhibition zone against L. monocytogenes Scott A, was selected for subsequent analysis. Both ribotyping and DNA sequencing of 16S ribosomal RNA gene demonstrated that the isolate was Lactococcus lactis subsp. lactis. Polymerase chain reaction and nucleotide sequencing revealed that the genomic DNA of the bean-sprout isolates contained a nisin Z structural gene. In MRS broth, bean-sprout isolate HPB 1688 survived at 3-4.5 degrees C for at least 20 d, grew at 4 degrees C and produced anti-listerial compounds at 5 degrees C. When co-cultured with L. monocytogenes in MRS broth, the isolate inhibited the growth of L. monocytogenes at 4 degrees C after 14 d and at 10 degrees C after 2 d. When co-inoculated with 10(2) cells g-1 of L. monocytogenes on fresh-cut ready-to-eat Caesar salad, L. lactis subsp. lactis (10(8) cells g-1) was able to reduce the number of L. monocytogenes by 1-1.4 logs after storage for 10 d at 7 zero and 10 degrees C. A bacteriocin-producing Enterococcus faecium was also able to reduce the numbers of L. monocytogenes on Caesar salad, but did not act synergistically when co-inoculated with L. lactis subsp. lactis.

Anti-Bacterial Agents↗
PubMed1997

Primary structure and functional expression of heavy- and light-chain variable region genes of a monoclonal antibody specific for human fibrin.

The immunoglobulin heavy- and light-chain variable region (VH and VK) genes were isolated from 8E5 hybridoma cells, which secreted monoclonal antibody against human fibrin by RT-PCR. An expression vector pOPE51-8E5 was constructed for the recombinant VH-VK scFv expression. The primary sequence of the variable regions was determined. Expression product was found in the periplasmic space and inclusion bodies by SDS-PAGE and immunoblotting. It was a 30 KDa single chain fragment (scFv) with the antigen-binding specificity of the parental monoclonal antibody. A light chain shuffling with an unspecific VL did not result in a loss of fibrin binding specificity.

Amino Acid Sequence↗
PubMed1997

Inhibitory effects of naturally occurring coumarins on the metabolic activation of benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene in cultured mouse keratinocytes.

Several naturally occurring coumarins to which humans are routinely exposed have been previously found to be potent inhibitors and inactivators of cytochrome P450 (P450) 1A1-mediated monooxygenase in both murine hepatic microsomes and in a reconstituted system using purified human P450 1A1 [Cai et al. (1993) Chem. Res. Toxicol., 6, 872-879 and Cai et al. (1996) Chem. Res. Toxicol., 9, 729-736]. In the present study, several of these coumarins were investigated for their inhibitory effects on the metabolism and metabolic activation of benzo[a]pyrene (B[a]P) and 7,12-dimethylbenz[a]anthracene (DMBA) in cultured mouse keratinocytes. Initial analysis of B[a]P metabolism in cultured keratinocytes showed that imperatorin, isoimperatorin, coriandrin, and bergamottin, at concentrations of 2 nM equal with B[a]P, reduced the formation of water-soluble metabolites of B[a]P by 33% to 57%. Bergamottin and coriandrin were the most potent inhibitors of the compounds examined. HPLC analysis of organic solvent-soluble metabolites of B[a]P indicated that all the coumarins tested significantly reduced the formation of individual B[a]P metabolites (including phenols, diols and tetraols). However, the greatest effect was on the formation of B[a]P tetraols. Additional experiments determined the ability of selected coumarins to block covalent binding of B[a]P and DMBA to DNA in keratinocytes. Bergamottin preferentially inhibited the binding of B[a]P to DNA by 56%, while coriandrin preferentially inhibited the binding of DMBA to DNA by 48%. Notably, analysis of individual DNA adducts formed from B[a]P and DMBA indicated that both bergamottin and coriandrin specifically inhibited the formation of anti diol-epoxide DNA adducts derived from both hydrocarbons. The preferential inhibitory effect of bergamottin and coriandrin on the formation of anti diol-epoxide adducts derived from DMBA was further confirmed by separation of anti- and syn-diol-epoxide-DNA adducts using immobilized boronate chromatography. The current study demonstrates that certain naturally occurring coumarins inhibited metabolic activation of B[a]P and DMBA in cultured mouse keratinocytes and specifically inhibited the formation of DNA adducts derived from the anti diol-epoxide diastereomers from either hydrocarbon. The current data also suggest that certain naturally occurring coumarins may possess anticarcinogenic activity toward polycyclic aromatic hydrocarbons.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1997

Effect of naturally occurring coumarins on the formation of epidermal DNA adducts and skin tumors induced by benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene in SENCAR mice.

Several naturally occurring coumarins previously found to be potent inhibitors of mouse hepatic ethoxyresorufin-O-deethylase (EROD) and/or pentoxyresorufin-O-dealkylase (PROD) were examined for their effects on formation of benzo[a]pyrene (B[a]P) and 7,12-dimethylbenz[a]anthracene (DMBA) DNA adducts in mouse epidermis, as well as, their effects on skin tumor initiation by these polycyclic aromatic hydrocarbons (PAH). Bergamottin, a potent inhibitor of hepatic EROD, given topically 5 min prior to an initiating dose of B[a]P, significantly decreased total covalent binding of B[a]P to DNA in a dose-dependent manner 24 h after treatment. A dose of 400 nmol bergamottin reduced covalent binding of B[a]P by 72%. Coriandrin, at a dose of 400 nmol also significantly reduced total covalent binding of B[a]P by 59%. In addition, formation of the major (+)anti-B[a]P-diol epoxide-N2-dGuo adduct was selectively reduced by both of these coumarins. In contrast, bergamottin and coriandrin did not significantly decrease covalent binding of DMBA to epidermal DNA at doses of either 400 nmol or 800 nmol. Imperatorin and isopimpinellin, which are more potent inhibitors of hepatic PROD activity, significantly reduced overall binding of DMBA to epidermal DNA by 67% and 52%, respectively, when applied at doses of 400 nmol. These two coumarins also inhibited B[a]P-DNA adduct formation at similar doses but to a lesser extent. Imperatorin at a dose of 400 nmol dramatically decreased formation of covalent DNA adducts derived from both the anti and syn diol epoxides of DMBA. Bergamottin was a potent inhibitor of tumor initiation by B[a]P while coriandrin was less effective in this regard. Imperatorin was an effective inhibitor of skin tumor initiation by DMBA and also inhibited complete carcinogenesis by this PAH. At dose levels higher than those effective against DMBA, imperatorin also inhibited tumor initiation by B[a]P. The results demonstrate that several naturally occurring coumarins possess the ability to block DNA adduct formation and tumor initiation by PAHs such as B[a]P and DMBA. The mechanism for reduced DNA adduct formation and tumor initiation appears to involve inhibition of the P450s involved in the metabolic activation of these hydrocarbons. Finally, the differential effects of certain coumarins on B[a]P vs DMBA DNA adduct formation and tumor initiation may be useful for dissecting the role of specific cytochromes P450 in their metabolic activation.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1997

Nitrogen deprivation of Anabaena sp. strain PCC 7120 elicits rapid activation of a gene cluster that is essential for uptake and utilization of nitrate.

A transposon bearing luxAB, encoding luciferase, as a reporter of transcription was used to identify genes that are activated rapidly upon deprivation of Anabaena sp. strain PCC 7120 of fixed nitrogen. The three transposon-marked loci that were identified as responding most rapidly and strongly are closely linked and situated within nirA and nrtC and between nrtD and narB, genes whose products are responsible for uptake and reduction of NO2- and NO3-. A strain bearing a transcriptional fusion of narB to luxAB was constructed. Luminescence catalyzed by LuxAB was used to report on the expression of the interrupted genes. Whether these genes are regulated only coordinately is discussed.

Amino Acid Sequence↗
PubMed1997

Mechanisms of onset responses in octopus cells of the cochlear nucleus: implications of a model.

The octopus cells of the posteroventral cochlear nucleus receive inputs from auditory-nerve fibers and form one of the major ascending auditory pathways. They respond to acoustic and electrical stimulation transiently and are believed to carry temporal information in the precise timing of their action potentials. The mechanism whereby onset responses are generated is not clear. Proposals aimed at elucidating the mechanism range from neural circuitry and/or inhibition, "depolarization block" (or inactivation of Na+ channels), and the involvement of a 4-aminopyridine (4-AP)-sensitive, low-threshold channel (K(LT)). In the present study, we used a compartment model to investigate possible mechanisms. The model cell contains a soma, an axon, and four passive dendrites. Four kinds of ionic channels were included in the soma compartment: the Hodgkin-Huxley-like Na+ and K+ channels, a 4-AP-sensitive, low-threshold channel, K(LT), and a Cs+-sensitive, hyperpolarization-activated inward rectifier, Ih. DC currents and half-wave-rectified sine waves were used as stimuli. Our results showed that an onset response can be generated in the absence of neuronal circuitry of any form, thus suggesting that the onset response in octopus cells is regulated intrinsically. Among the many factors involved, low-input impedance, partly contributed by Ih, appears to be essential to the basic onset response pattern; also, the K(LT) conductance plays a major role, whereas the inactivation of Na+ channels probably plays only a secondary role. The dynamics of Ih also can modify the response pattern, but due to its slow kinetics, its role is probably limited to longer-term regulation under the conditions simulated in this study.

Acoustic Stimulation↗