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C Su

Publications and source records attributed to C Su.

At least 37 records · Page 2Linked to original sources

Evolutionary dynamics of the T-cell receptor VB gene family as inferred from the human and mouse genomic sequences.

The diversity of T-cell receptors is generated primarily by the variable-region gene families, each of which is composed of a large number of member genes. The entire genomic sequence of the variable region (VB) of the T- cell receptor beta chain from humans and mice has become available. To understand the evolutionary dynamics of the VB gene family, we conducted a phylogenetic analysis of all VB genes from humans and mice, as well as a detailed analysis of internal DNA duplications in the human genomic VB region. The phylogenetic tree obtained shows that human and mouse VB genes intermingle extensively rather than forming two separate clusters and that many gene duplications occurred both before and after the divergence between primates and rodents. Analyzing the genomic maps of transposable elements (e.g., LINEs and SINEs) and relic VB genes in the VB gene region, we present evidence that a 20-kb VB region duplicated tandemly four times in the human lineage during the last 32 Myr, and 6 out of the 15 VB genes in this region have become nonfunctional during this period. Our results show that the VB gene family is subject to evolution by a birth-and-death process rather than to concerted evolution.

Amino Acid Sequence↗

Cancer preventive effect of Morinda citrifolia (Noni).

Morinda citrifolia (Noni) has been extensively used in folk medicine by Polynesians for over 2,000 years. It has been reported to have broad therapeutic effects, including anticancer activity, in both clinical practice and laboratory animal models. The mechanism for these effects remains unknown. The hypothesis that Morinda citrifolia possesses a cancer preventive effect at the initiation stage of carcinogenesis was studied. Our preliminary data indicated that 10% Tahitian Noni Liquid Dietary Supplement or Tahitian Noni Juice (TNJ), made from Morinda citrifolia fruit by Morinda Inc, in drinking water for one week was able to prevent DMBA-DNA adduct formation. The levels of DMBA-DNA adducts were reduced by 30% in the heart, 41% in the lung, 42% in the liver, and 80% in the kidney of female SD rats. Even more dramatic results were obtained in male C57 BL-6 mice: 10% TNJ was able to reduce DMBA-DNA adduct formation by 60% in the heart, 50% in the lung, 70% in the liver, and 90% in the kidney. In order to explore the mechanism of this preventive effect, the antioxidant activity of TNJ was examined in vitro by lipid hydroperoxide (LPO) and tetrazolium nitroblue (TNB) assays. In the LPO assay, LPO oxidizes leucomethylene blue to methylene blue in the presence of hemoglobin. The resultant blue color was quantified at 660 nm spectrophotometrically. In the TNB assay, superoxide anion radicals (SAR) reduce TNB into formazan blue that was also measured by absorption at 602 nm. TNJ showed a dose-dependent inhibition of both LPO and SAR in our system. The antioxidant activity of TNJ was compared to the effects of vitamin C, grape seed powder (GSP), and pycnogenol (PYC) at the daily dose per serving level recommended by U.S.RDAs or manufacturers. The results suggest that prevention of carcinogen-DNA adduct formation and the antioxidant activity of TNJ may contribute to the cancer preventive effect of Morinda citrifolia.

9,10-Dimethyl-1,2-benzanthracene↗

Staphylococcus aureus agr genotypes with enterotoxin production capabilities can resist neutrophil bactericidal activity.

Staphylococcus aureus pathogenicity is mainly due to the production of a number of secreted and cell surface-associated proteins under the regulation of the agr gene. A region of the agr gene was used to subgroup S. aureus strains according to restriction fragment length polymorphisms. Additionally, strains were subtyped according to the coagulase gene in order to strengthen discriminatory power. Virulence capabilities of agr genotype subgroups were evaluated using an in vitro neutrophil bactericidal assay, which showed that prevalent genotypes were significantly better at evading this primary host defense. Multiplex PCR was then used to detect enterotoxin genes among the genotype subgroups in order to determine possible virulence candidates that enable strains to combat neutrophil killing. The prevalent genotype strains were found to possess higher production capabilities for enterotoxin A than did low-prevalence strains. The significance of enterotoxin A production capabilities in affecting pathogenicity of S. aureus strains was evaluated and found to have a profound effect on neutrophil killing abilities. The use of a large epidemiological database as a tool for subgrouping strains with varying degrees of pathogenicity has allowed the identification of relevant and previously undefined virulence factors that affect a pathogen's capability to overcome host immune defenses.

Animals↗

High-resolution typing of Toxoplasma gondii using microsatellite loci.

High-resolution typing of Toxoplasma gondii is essential to understand the effect of genetic differences among strains on the variation in disease manifestation and transmission patterns. Current typing methods discern 3 lineages with minimal within-lineage variation. Described here are 6 new variable loci. These loci, including a minisatellite and 5 microsatellites, were more polymorphic than allozymes, restriction fragment length polymorphisms, and sequence variation in introns. Most importantly, these loci revealed, for the first time, substantial within-lineage variation that was over 6-fold higher than that detected by other markers. Genotyping at these loci facilitates classification of isolates beyond the lineage level.

Animals↗

The steroid receptor antagonists RU40555 and RU486 activate glucocorticoid receptor translocation and are not excreted by the steroid hormones transporter in L929 cells.

RU40555 is a recently available glucocorticoid receptor (GR) antagonist that differs from RU486 by a methyl radical. We have used the mouse fibroblast cell line L929 to study the in vitro effects of RU40555 on GR translocation and function and on the membrane steroid hormones transporter. The results showed that: 1) RU40555 competed for the binding of labelled dexamethasone (Dex) with a K(i) of 2.4 nM; 2) both RU40555 and RU486 were equally potent inhibitors of Dex-induced GR-mediated gene transcription; 3) maximum GR translocation induced by micromolar concentrations of Dex and the GR antagonists was approximately 30-55% loss in the cytoplasmic GR and approximately 40-90% increase in the nuclear GR (assessed by GR immunostaining in cytoplasm and nucleus and western blots of immunoprecipitated GR protein in cytosolic and nuclear fractions) and was similar for the two antagonists; 4) at nanomolar concentrations, RU40555 and RU486 induced more GR translocation than Dex (assessed by [(3)H]Dex binding and western blot of immunoreactive GR in the same cytosolic homogenates); 5) blocking the steroids membrane transporter with verapamil (100 microM) in the presence of Dex (10 nM) increased GR translocation to levels similar to those induced by RU40555 (10 nM) and RU486 (10 nM) alone; 6) verapamil did not affect GR translocation in the presence of RU40555 or RU486. These data demonstrate similar quantitative effects on GR translocation by RU486 and the new GR antagonist, RU40555. Moreover, RU40555, like RU486, is an effective GR antagonist. Finally, there is no evidence that the intracellular concentrations of RU40555 or RU486 are regulated by the steroids membrane transporter in L929 cells.

Analysis of Variance↗

Studies on the development of DNA vaccine against Cysticercus cellulosae infection and its efficacy.

DNA vaccine against Cysticercus cellulosae infection was developed and its efficacy was tested. A pair of primers specific to antigen B gene of C. cellulosae was designed which amplified the gene successfully with RT-PCR. The gene was ligated to PV93 vector, and the recombinant of antigen B gene and PV93 was transformed to JM83 cells. The transformed JM83 cells were cultured in a large scale and the plasmid purified. Based on the recombinant plasmid. a DNA vaccine was developed and used to vaccinate two groups of experimental pigs. In each group, there was a routine vaccine, an enhanced vaccine and a control group. Groups 1 and 2 were challenged at 4 months and at 14 days post vaccination respectively with eggs of Taenia solium. The antibody response was also tested with ELISA. The results suggested that all animals vaccinated AgB gene DNA vaccine, no matter by routine or enhanced vaccine, their antibodies reached maximum peak 23 days post vaccination and decreased gradually. When the animals were challenged 4 months after vaccination, they had strong immunity and the parasites decrease rates were 91.2% and 93.1% respectively. When pigs vaccinated with AgB gene DNA vaccine were challenged 14 days post vaccination with 18,000 eggs/pig. The animals showed strong immunity and the parasite decrease rates were 99.5% and 84.9% respectively. However at that time, the antibodies did not reach the peak. While in the control group, the number of C. cellulosae was as many as 2,500. It was concluded that the pigs vaccinated with DNA vaccine had strong immunity against infection of eggs of T. solium.

Animals↗

[Analysis of the mitochondria-related protein of Schistosoma japonicum and its antigen epitopes].

OBJECTIVE: To sequence the cloned gene Sj338 and to identify the encoded protein and its antigen epitopes. METHODS: The Sj338 gene fragment obtained from adult S. japonicum cDNA library amplified by PCR method was subcloned into pGEM-T vector for sequencing. The sequence of nucleotides and the characteristics of the encoded protein were analyzed by DNASIS Program and Goldkey DNA and Protein Analytical Program, and then the homology of the amino acid sequence was searched on the BLAST net. RESULTS: The cloned rSj338 gene was demonstrated to be 487 bp containing one 459 bp ORF, encoding a protein consisted of 153 amino acids with a molecular weight of 17.6 kDa. The amino acid sequence of the recombinant protein rSj338 shared 46% identity with that of the corresponding part of human mitochondrial import receptor and 44% identity with that of the Rattus sp. mitochondrial precursor receptor. The possible antigen epitopes were predicted within the peptide fragments of 26-32 aa, 37-46 aa and 147-151 aa. CONCLUSION: The protein encoded by rSj338 gene fragment might be the mitochondria-related protein of Schistosoma japonicum.

Amino Acid Sequence↗

[Clonning and identification of the gene encoding specific IgE antibody-related antigen of Schistosoma japonicum].

OBJECTIVE: To obtain the gene encoding specific IgE-related antigen of Schistosoma japonicum from the cDNA library of adult Schistosoma japonicum. METHODS: The pooled sera from 15 individuals with high levels of specific IgE antibody against SWAP were absorbed with Protein-G and used for screening of IgE-related antigen from the adult worm cDNA library of Schistosoma japonicum. The inserted cDNA was amplified by PCR and sequenced. According to the first reading frame of the sequence, a pair of new probes, in which EcoR I and Not I sites were incorporated respectively, were designed and used to amplify the target gene. Then, the gene was cloned into vector pGEM-T and subcloned into expression vector pGEX-6p-1. The fusion protein was expressed, analysed by SDS-PAGE and identified by Western blotting with the specific IgE antibody, respectively. RESULTS: The inserted cDNA fragment from the positive clone was about 1,200 bp, with the ORF of 507 bp which encoded 169 amino acids. The deduced molecular weight of the recombinant protein was 19.3 kDa. The homology between the target gene (Sj43B) and other known DNA sequences was less than 40%. The fusion protein expressed by the recombinant vector pGEX-6p-1/Sj43B could be recognized by schistosome specific IgE antibody. CONCLUSION: Sj43B may encode the specific IgE-related antigen of Schistosoma japonicum. The successful construction of recombinant plasmid pGEX-6p-1/Sj43B lay the groundwork for further studies on immunological characteristics and protection immunity of the recombinant protein.

Animals↗

[Chemical constituents from Salix oritrepha Schneid].

OBJECTIVE: To study chemical constituents of the above ground part of Salix oritrepha. METHOD: Using chromatography to isolate compounds and their structure were identified by physical, chemical and spectral techniques. RESULT: Four compounds were isolated and elucidated as beta-sitosterol, beta-sitosteryl-3-O-glucoside, friedelin, luteolin-7-O-glucoside. CONCLUSION: All of them were isolated from S. oritrepha for the first time. Friedelin from the plants of genus Salix for the first time.

Flavonoids↗

[Research on the near-infrared (NIR) photon migration in multi-layered structures of biological tissues].

The phenomena of photon migration in multi-layered structures of biological tissue is very important in the field of non-invasive near-infrared optical diagnosis. In this paper, a two-layered structure consisting of the fat and the muscle of poke was used as the sample. Near-infrared light source and detector were located on the surface of the fat and a part of the photons reflected by the two-layered structure of the tissue were detected. The relationship between the optical current and the distance from source to detector and the relationship between the optical current and the thickness of the fat layer were given. In order to analyze the experiment results, Monte-Carlo simulation was used to calculate the photons migration traces in the uniform tissues and the Fresnel's law is used to determine the refraction and reflection components of a photon package at the boundary between the fat and the muscle. The results showed that the simulation data coincident well with the experimental results. The conclusion is that the experiment and the simulation method provided in this study may be useful to analyze the photon migration in a mult-layered tissue and to optimize the design of the transducer.

Adipose Tissue↗

Analyzing high-density oligonucleotide gene expression array data.

We have developed methods and identified problems associated with the analysis of data generated by high-density, oligonuceotide gene expression arrays. Our methods are aimed at accounting for many of the sources of variation that make it difficult, at times, to realize consistent results. We present here descriptions of some of these methods and how they impact the analysis of oligonucleotide gene expression array data. We will discuss the process of recognizing the "spots" (or features) on the Affymetrix GeneChip(R) probe arrays, correcting for background and intensity gradients in the resulting images, scaling/normalizing an array to allow array-to-array comparisons, monitoring probe performance with respect to hybridization efficiency, and assessing whether a gene is present or differentially expressed. Examples from the analyses of gene expression validation data are presented to contrast the different methods applied to these types of data.

Base Sequence↗

Potentiation of ganodermic acid S on prostaglandin E(1)-induced cyclic AMP elevation in human platelets.

Ganodermic acid S (GAS), isolated from the Chinese medicinal fungus Ganoderma lucidum (Fr.) Karst (Polyporaceae), exhibits inhibitory effects on platelet responses to various aggregating agonists. Our study demonstrated that GAS also participated in potentiating the response of human gel-filtered platelets to prostaglandin (PG) E(1). GAS at <20 microM did not show any significant change of basal cyclic AMP level in gel-filtered platelets. However, GAS potentiated the PGE(1)-evoked cyclic AMP level in a bell-shaped, concentration-dependent manner. The agent at 7.5 microM enhanced the level up to 1.8-fold of that evoked by PGE(1) alone. Collagen did not inhibit the PGE(1)-induced cyclic AMP level in platelets pretreated with GAS at 6 to 7.5 microM. In the presence of 7.5 microM GAS, the agent enhanced the inhibition of PGE(1) on platelet response to collagen in: phosphorylation of myosin light chain and pleckstrin; alpha-granule secretion; cell aggregation and protein-tyrosine phosphorylation. In addition, the agent along with PGE(1) almost abolished the dense-granule secretion and thromboxane (TX) B(2) formation. The results suggest that GAS played an additional role in potentiating the PGE(1)-induced cyclic AMP synthesis. GAS and PGE(1) inhibited additively the platelet response to collagen.

Alprostadil↗

Kinetics of manganese lipoxygenase with a catalytic mononuclear redox center.

Manganese lipoxygenase was isolated from the take-all fungus, Gaeumannomyces graminis, and the oxygenation mechanism was investigated. A kinetic isotope effect, k(H)/k(D) = 21-24, was observed with [U-(2)H]linoleic acid as a substrate. The relative biosynthesis of (11S)-hydroperoxylinoleate (11S-HPODE) and (13R)-hydroperoxylinoleate (13R-HPODE) was pH-dependent and changed by [U-(2)H]linoleic acid. Stopped-flow kinetic traces of linoleic and alpha-linolenic acids indicated catalytic lag times of approximately 45 ms, which were followed by bursts of enzyme activity for approximately 60 ms and then by steady state (k(cat) approximately 26 and approximately 47 s(-1), respectively). 11S-HPODE was isomerized by manganese lipoxygenase to 13R-HPODE and formed from linoleic acid at the same rates (k(cat) 7-9 s(-1)). Catalysis was accompanied by collisional quenching of the long wavelength fluorescence (640-685 nm) by fatty acid substrates and 13R-HPODE. Electron paramagnetic resonance (EPR) of native manganese lipoxygenase showed weak 6-fold hyperfine splitting superimposed on a broad resonance indicating two populations of Mn(II) bound to protein. The addition of linoleic acid decreased both components, and denaturation of the lipoxygenase liberated approximately 0.8 Mn(2+) atoms/lipoxygenase molecule. These observations are consistent with a mononuclear Mn(II) center in the native state, which is converted during catalysis to an EPR silent Mn(III) state. We propose that manganese lipoxygenase has kinetic and redox properties similar to iron lipoxygenases.

Ascomycota↗

Expression of human mitochondrial thymidine kinase in Escherichia coli: correlation between the enzymatic activity of pyrimidine nucleoside analogues and their inhibitory effect on bacterial growth.

Mitochondrial thymidine kinase (TK2) phosphorylates pyrimidine nucleosides to monophosphates and is expressed constitutively through the cell cycle in all cells. Because of the overlap of its substrate specificity with that of the cytosolic thymidine kinase (TK1) and deoxycytidine kinase (dCK), it has been difficult to determine the role of TK2 in activating nucleosides used in chemotherapy. In this report, we described the construction of a recombinant Escherichia coli strain which could be used to test if TK2 activity is limiting for the toxicity of nucleosides. Enzymes of bacterial origin which are involved in thymidine and deoxyuridine anabolism and catabolism were eliminated, and the cDNA for human TK2 was introduced. In the crude extract of the engineered E. coli, the level of thymidine kinase was, after induction of TK2 expression, several hundred fold higher than in the control strain. Several pharmacologically interesting nucleoside analogues, including 3'-azidothymidine, 2',3'-didehydro-2',3'-dideoxythymidine, and 2', 3'-dideoxy-beta-L-3'-thiacytidine, were tested for their effects on the growth of this recombinant strain. For a comparison, the phosphorylation of these compounds was determined with purified recombinant TK1, TK2, and dCK. A correlation was observed between the phosphorylation of several of these compounds by TK2 and their effects on bacterial growth. These results demonstrate that activation of growth-inhibiting pyrimidine nucleosides can be catalyzed by TK2, and together with recombinant E. coli strains expressing other cellular nucleoside kinases, this whole-cell bacterial system may serve as a tool to predict the efficacy and side effects of chemotherapeutic nucleosides.

Cell Division↗

Glial cell line-derived neurotrophic factor in the rat pituitary gland.

The presence of glial cell line-derived neurotrophic factor (GDNF) is described within specific regions of the adult rat pituitary gland. Immune staining methods revealed a small number of GDNF-immunopositive cells in the anterior lobe, and in areas of the neural lobe, while no immunoreactive endocrine cells were observed in the intermediate lobe. In the neural lobe, immunofluorescence methods were also used to demonstrate that GDNF and glial fibrillary acidic protein (GFAP) are co-localized in the glial cells (pituicytes) of the neural lobe. GDNF was not co-localized with neurofilament (NF) in nerve fibers of the neural lobe, suggesting that it is not present in axonal fibers. Measurements of GDNF content in separated anterior and neurointermediate lobes were also performed, using an enzyme-linked immunoassay (ELISA). Values for GDNF were slightly higher in the neurointermediate lobe than those obtained for the anterior lobe. The presence of GDNF in areas of the pituitary is discussed in the context of its possible function to support and maintain hypothalamic innervation, as well as a potential autocrine factor within endocrine cells.

Animals↗

Phylogenetic relationships of Staphylococcus aureus from bovine mastitis based on coagulase gene polymorphism.

Coagulase gene restriction fragment length polymorphism (RFLP) patterns were analyzed to determine the phylogenetic relationship among isolates of Staphylococcus aureus from the Czech Republic (n = 27), France (n = 48), Korea (n = 115) and the United States (n = 278). A total of 468 isolates of S. aureus were subtyped into 41 coagulase genotypes. Cluster analysis placed the 41 types into nine clusters. Eighteen API Staph profiles were determined for 102 S. aureus isolates representing 1 to 4 isolates of each coagulase type. The results of the study suggest that based on coagulase gene RFLP analysis, several genetic variants of S. aureus are prevalent. Comparison of coagulase and API Staph profiles indicated that the two identification system were independent of each other.

Animals↗

[Correlation of inactivation of CDKN2/p16 gene by methylation and lung cancers].

OBJECTIVE: To study the relationship between methylation state of CDKN2/p16 gene and lung cancers. METHODS: Genomic DNA was digested with methylation-sensitive enzymes and Southern blotting was used to analyses the methylation state of CDKN2/p16 gene in 89 cases of lung cancer. RESULTS: Twenty-one out of 89 lung cancers(23.6%) shown methylation of CDKN2/p16 gene, and 17 of them occurred in 42 P16 negative cases with a rate of 40.5%(17/42). CONCLUSION: Methylation of CDKN2/p16 gene 5' CpG island is probably an important mechanism of gene inactivation; it may take part in the onset and process of lung cancer.

Adult↗