Biomedical subjects

P Feng

Publications and source records attributed to P Feng.

At least 91 records · Page 5Linked to original sources

PubMed1997

[Identification of the anti-rIL-1ra McAb].

Using dot-immuno-blot and Western blotting techniques, we have identified the specificity of anti-rIL-1ra McAbs. The results showed that the anti-rIL-1ra McAb 2E4 specifically reacted with rIL-1ra, from which the recognizing signal of dot-immunoblot was significantly positive (+2) and a single rIL-1ra protein band (20 kd) was clearly recognized by Western blotting. The control results of PcAb recognization have further proved the high sensitivity of Western blotting and high specificity of rIL-1ra McAb 2E4. The successful identification of rIL-1ra McAb 2E4 will be helpful to further understand the biologic effects, especially the pro-inflammatory properties, to study the change of rIL-1ra level during inflammation, and to instruct the clinical use of rIL-1ra.

Animals↗

PubMed1997

[Primary study of bacterial adhesion of prosthetic valve materials].

The adhesion of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans to Dacron and pyrolite carbon was measured in vitro with plate counting. The experimental result showed: the bacterials used in the experiment have adhesion to prosthetic valve materials; the bacterials have stronger adhesion to Dacron than to Pyrolite carbon; the bacterial adhesive capacity is significantly different and is affected by mechanical and chemical nature of biomaterials; Staphylococcur aureus has the strongest adhesion to Dacron, and Pseudomonas aeruginosa has the strongest adhesion to pyrolite carbon in all bacterials.

Bacterial Adhesion↗

PubMed1996

Sodium butyrate is a potent modulator of smooth muscle cell proliferation and gene expression.

Sodium butyrate is a small, naturally occurring molecule with demonstrated activity on cell growth and differentiation. However, its effect on smooth muscle cells had not been examined. We have found that sodium butyrate and its more stable in vivo analogue tributyrin are potent DNA synthesis and cell proliferation inhibitors. The inhibitory activity of sodium butyrate was not mediated by an elevation of endogenous cAMP levels, a known pathway involved in SMC growth-arrest and maintenance of the contractile phenotype. Consistent with the concept that its activity is mediated by a cAMP-independent pathway, butyrate was able to augment the maximum DNA synthesis inhibitory effect of various agents that elevated intracellular cAMP levels. Additionally, butyrate present for just the initial 8 h or present as late as 16 h after serum addition was able to inhibit DNA synthesis. By contrast, the cAMP analogue, 8Br-cAMP had to be present throughout the entire G0 to S phase of the cell cycle to effectively inhibit DNA synthesis. These results indicated that sodium butyrate inhibited SMC growth through a cAMP-independent mechanism. We also found that sodium butyrate was unable to abrogate the expression of the serum-inducible genes c-fos, c-myc, Ki-Ras and PS4, but was able to directly stimulate the expression of PS4 and thrombospondin. These results indicate that a number of important early G1 events initiated by serum growth factors are unaltered by sodium butyrate and that this compound is able to directly stimulate the expression of certain genes normally associated with SMC proliferation.

Adenosine Monophosphate↗

PubMed1996

Reversal of TR-T3 inhibition of the hTRH gene by excess TR ligand-binding domain: evidence for novel accessory protein.

The thyrotropin-releasing hormone (TRH) gene is regulated negatively at the transcriptional level by thyroid hormone (T3). T3 positive regulatory effects on other target genes, such as the growth hormone gene, are mediated through heterodimerization of thyroid hormone receptors (TRs) with RXR or other auxiliary nuclear protein(s). To explore whether an accessory co-suppressor protein(s) may be involved in T3 inhibitory regulation of human TRH gene transcription, transient gene expression studies have been carried out using a hTRH-luciferase (TRH-Luc) chimetric reporter construct, an hTR beta 1 expression construct, and pABgal-hTR beta 1 ligand-binding domain (LBD) fusion constructs, cotransfected into a human neuroblastoma cell line (HTB-11,ATCC). Results herein indicate that T3-dependent inhibitory regulation (48-60% of control) of the hTRH gene promoter by hTR beta 1-T3 complexes could be abrogated completely by cotransfection of a 10 x excess of hTR beta 1-LBD (TR 168-456 aa) in a pABgal94 vector. In striking contrast, cotransfection of a 10 x excess of highly truncated hTR beta 1-LBD (TR 452-456 aa) failed to reverse T3-mediated TRH promoter inhibition. This squelching effect by excessive intact TR-LBD, moreover, could not be reversed by raising T3 concentration 100-fold (from 10(-8) to 10(-6) M), thus excluding a squelching effect of T3 itself by excess LBD. These results suggest that negative regulation of the hTRH gene promoter activity by TR beta 1-T3 complexes involves interactions with an accessory co-suppressor protein, which may bridge DNA-bound TR beta 1-T3 complexes to the transcriptional initiation complex.

Binding Sites↗

PubMed1996

Characterization of nonmotile variants of Escherichia coli O157 and other serotypes by using an antiflagellin monoclonal antibody.

An antiflagellin monoclonal antibody (15D8) was used to detect the presence of flagella in nonmotile variants of several pathogenic Escherichia coli serotypes. Of the 48 isolates examined, 15 reacted with monoclonal antibody 15D8 and were culturally confirmed to be motile. Of the 38 clinical strains designated O157:NM or O157:H-, 7 were antibody reactive and motile and agglutinated with anti-H7 sera.

Antibodies, Monoclonal↗

PubMed1996

Effect of direct-fed fibrolytic enzymes on the digestive characteristics of a forage-based diet fed to beef steers.

A study was conducted to examine the method of delivery of a solution containing cellulases and xylanases on the digestion of a forage-based diet. Five ruminally cannulated beef steers (536 kg BW) were randomly assigned to a control (CON) or one of four enzyme treatments in a 5 x 5 Latin square experiment. Steers were fed a 70:30 (DM basis) grass hay:barley diet. Enzyme-treated rations contained a solution of fibrolytic enzymes at the rate of 1.65 mL/kg of forage DM. Enzyme application treatments were 1) enzyme to forage 24 h before feeding (F-24), 2) enzyme to forage 0 h before feeding (F-0), 3) enzyme to barley 0 h before feeding (B-0), and 4) enzyme infused ruminally 2 h after feeding (RI). Dry matter and NDF intakes were not different (P > .10) across treatments. Ruminal pH was lower and total VFA concentration at 16 h after-feeding was greater (P < .10) for steers fed enzyme treatments compared with CON. Rate of NDF disappearance was greater (P < .05) for enzyme-treated than for untreated grass substrate. Ruminal infusion of enzymes compared with F-24 and F-0 produced lower disappearance of DM at 8 and 32 h (P < .10), NDF at 32 h (P < .10), and DM and NDF at 96 h (P < .05). Rate of DM disappearance of enzyme-treated grass hay was greater (P < .10) for steers fed B-0 than for those fed F-24 and F-0 and for CON than for F-24 and F-0. Total tract digestibility of DM, NDF, and ADF was greater (P < .10) for F-24 and F-0 than for CON. Forage transit time was shorter (P < .10) for B-0 than for F-24 and F-0; however, all other contrasts for particulate passage did not differ (P > .10). Results from this study indicate that direct application of enzymes to forages is capable of improving forage digestion.

Animals↗

PubMed1996

Effect of enzyme preparations on in situ and in vitro degradation and in vivo digestive characteristics of mature cool-season grass forage in beef steers.

In situ and in vitro studies with a 3 x 2 x 5 factorial arrangement of treatments with an added untreated control evaluated three enzyme preparations, two levels of enzyme, and five moisture conditions of grass forage. Enzyme preparations predominantly contained cellulase and xylanase and will be designated as enzyme 1 (E1), enzyme 2 (E2), and a 50:50 combination of E1 and E2 (E1E2). The five moisture conditions included fresh, wilted, dried and rehydrated to fresh, dried and rehydrated to wilt, and dried grass. Addition of the high level of E1E2 to dried grass improved (P < .05) in vitro DM (43.5 vs 38.7%) and NDF (31.1 vs 26.0%) disappearance (48 h incubation) compared with the control treatment. Also, IVDMD was greater (P < .05) for the low level of E1 applied to wilted grass compared with the control. No other enzyme application improved in situ or in vitro disappearance of substrate over the control. In vivo responses of enzyme treatments found most likely to be effective from degradability studies were measured using four ruminally cannulated steers in a 4 x 4 Latin square experiment. Treatments examined were E1 applied to fresh forage, then dried; E1 applied to wilted forage, then dried; E1E2 applied to dry forage immediately before feeding (E-dry), and untreated forage (control). All forage treatments were harvested as dry hay. Total diet and hay DM intakes were greater (P < .05) for the E-dry than for the control diet. Rate of in situ NDF disappearance and total tract DM and NDF digestibility were greater (P < .05) for the E-dry than for the other treatments. Ruminal fluid ammonia N concentration, total VFA concentration, and pH were not altered (P > .10) by dietary treatment. Ruminal particulate passage rate was greater (P < .05) and ruminal retention time was shorter (P < .05) for the E-dry than for the control treatment. Data from this study suggest that addition of fibrolytic enzymes to grass hay before feeding has the potential to enhance intake and digestion.

Analysis of Variance↗

PubMed1996

The glutathione S-transferase M1 (GSTM1) null genotype and benzidine-associated bladder cancer, urine mutagenicity, and exfoliated urothelial cell DNA adducts.

Multiple studies in the general population have suggested that subjects with the glutathione S-transferase M1 (GSTM1)-null genotype, who lack functional GSTM1, are at higher risk for bladder cancer. To evaluate the impact of the GSTM1-null genotype on bladder cancer caused by occupational exposure to benzidine and to determine its influence on benzidine metabolism, we carried out three complementary investigations: a case-control study of bladder cancer among workers previously exposed to benzidine in China, a cross-sectional study of urothelial cell DNA adducts and urinary mutagenicity in workers currently exposed to benzidine in India, and a laboratory study of the ability of human GSTM1 to conjugate benzidine and its known metabolites in vitro. There was no overall increase in bladder cancer risk for the GSTM1-null genotype among 38 bladder cancer cases and 43 controls (odds ratio, 1.0; 95% confidence interval, 0.4-2.7), although there was some indication that highly exposed workers with the GSTM1-null genotype were at greater risk of bladder cancer compared to similarly exposed workers without this allele. However, the GSTM1 genotype had no impact on urothelial cell DNA adduct and urinary mutagenicity levels in workers currently exposed to benzidine. Furthermore, human GSTM1 did not conjugate benzidine or its metabolites. These results led us to conclude that the GSTM1-null genotype does not have an impact on bladder cancer caused by benzidine, providing a contrast to its association with elevated bladder cancer risk in the general population.

Benzidines↗

PubMed1996

[Labeling DNA probe by polymerase chain reaction].

The use of polymerase chain reaction (PCR) for labeling probe has been demonstrated to offer various advantages including efficient labeling of DNA fragments as small as 72 bp, direct labeling of genomic DNA, and labeling with subnanogram amounts of input DNA. Therefore, a procedure for the nonradioactive labeling of chromasomal DNA 203 bp fragments of Helicobacter pylori with the hapten digoxigenin (Dig) by PCR has been developed. The results showed that the concentration of probe labeled by PCR was much higher than that by random primer labeled. PCR has the advantage of rapidity and economy. It is a very effective technique for synthesis of Dig-labeled DNA probe.

DNA Probes↗

PubMed1995

Thyroid hormone enhancement of estradiol stimulation of breast carcinoma proliferation.

Thyroid hormone (T3) and estradiol (Est) modulate biological processes by binding to nuclear receptor proteins that, through interactions with specific response elements in the regulatory regions of genes, modulate gene transcription. Est stimulation of estrogen receptor (ER)-positive breast carcinoma cell growth occurs through its ability to bind to the ER and activate gene transcription. We now report that physiological concentrations of T3 significantly enhance Est stimulation of growth of a number of human breast carcinoma cell lines. The effect of T3 is specific for Est stimulation of growth and has no effect on insulin-like growth factor-I stimulation of growth. The effect of T3 on enhancing Est-mediated growth was specifically blocked by the addition of ligands inducing retinoid X receptor (RXR) homodimer receptor formation, suggesting that RXR-thyroid nuclear receptor (TR) heterodimer formation is required for the T3-mediated effect on estradiol-stimulated growth. Four thyroid nuclear receptors have been described in tissues, TR alpha 1, alpha 2, beta 1, and beta 2. Breast carcinoma cells were found to express TR beta 1 and TR alpha 2 mRNA and very low levels of TR alpha 1 mRNA. T3 did not increase ER mRNA or protein levels and did not enhance Est-mediated increases in gene transcription of a number of genes, i.e., transforming growth factor-alpha and pS2 which contain estrogen-response elements (EREs) in their regulatory regions. However, T3 enhanced Est-stimulated ERE-TK-CAT activity. Thus significant cross-talk appears to occur between the TRs and ER and T3 appears to enhance Est-mediated gene transcription.

Blotting, Northern↗

PubMed1995

Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR.

Mismatch amplification mutation assay primers, specific for a unique base substitution in uidA of Escherichia coli O157:H7, was coupled with primers for the Shiga-like toxin I (SLT-I) and SLT-II genes in a multiplex PCR assay. Analysis of 108 bacteria showed that all Escherichia coli serotype O157:H7 strains were identified simultaneously with the SLT types encoded by these strains.

Bacterial Toxins↗

PubMed1995

Isolation and characterization of a beta-D-glucuronidase-producing strain of Escherichia coli serotype O157:H7 in the United States.

A phenotypic variant of Escherichia coli serotype O157:H7 (G5101) was isolated from a patient with bloody diarrhea. Strain G5101 does not ferment sorbitol but is beta-D-glucuronidase and urease positive. Serotyping and colony hybridization using a serotype-specific DNA probe confirmed that the isolate was O157:H7. G5101 produces Shiga-like toxins I and II and contains an eae gene that is highly conserved in the O157:H7 serotype. This strain would have been missed by laboratories that screen for the sorbitol-negative, beta-D-glucuronidase-negative phenotype in isolating E. coli O157:H7 from clinical and food specimens.

DNA Probes↗

PubMed1995

Effect of barley variety and dietary barley content on digestive function in beef steers fed grass hay-based diets.

Five ruminally and duodenally cannulated steers were used in a 5 x 5 Latin square design with a 2 x 2 + 1 arrangement of treatments to study the effects of barley variety and dietary barley content on digestive function in steers fed grass hay-based diets. Barley varities evaluated were Russell and Steptoe, which had bulk densities of 67.7 and 64.5 kg/hL, respectively. Supplemental treatments were as follows: corn, low Russell (Ru-lo), low Steptoe (St-lo), high Russell (Ru-hi), and high Steptoe (St-hi). Corn, Ru-lo, and St-lo were provided at 30% (DM basis) of grass hay-based diets, whereas Ru-hi and St-hi were provided at 35.5% of diet DM (equal starch content as the corn treatment). No treatment differences (P > .10) were observed for DMI, ruminal particulate passage rate, and NDF total tract digestibility. Ruminal DM and starch digestibility were greater (P < .01) for barley-containing diets than for the corn diet. Similarly, total tract DM (P < .10) and starch (P < .01) digestibility was greater for barley than for corn diets. Microbial protein and non-NH3 N flow to the small intestine were greater (P < .01) for the barley diets than for the corn diet. Starch intake (P < .01) and DM digestibility (P < .10) were greater for high- than for low-barley diets; however, differences due to barley variety were not observed (P > .10). In situ disappearance of grass hay NDF at 8 and 96 h of incubation was greater (P < .05) for barley than for corn diets. Rate of in situ disappearance of grain DM was greater (P < .01) for barley than for corn and for Russell than for Steptoe barley. Responses suggest that ruminal and total tract digestibility and protein flow to the small intestine can be increased with barley compared with corn as an energy supplement to grass hay-based diets.

Animals↗

PubMed1995

A study of growth kinetics of the flocs from Schizosaccharomyces pombe.

Inducing yeast cells to self-flocculate could be considered a cell immobilization method. The growth kinetics of the flocs from Schizosaccharomyces pombe was studied in an experimental suspended-bed bioreactor with starch hydrolysate obtained by two-stage enzymatic hydrolysis. It was discovered that a limited oxygen supply was necessary during continuous ethanol fermentation with yeast flocs. The oxygen supplied was a kind of limited substrate effecting on the floc growth. Further, a kinetic model describing this floc growth was proposed.

Cell Division↗

PubMed1995

[Research on the different blood types umbilical cord blood hematopoietic progenitors mixed culture in vitro].

Using single layer agar culture method, we make a study on culture of human hematopoietic progenitor cell from umbilical cord blood. The result showed the mean value of the granulocyte macrophage progenitor cells (CFU-GM) from twenty neonates cord blood was 95.6 +/- 36.5/10(6) MNC. We compared the mixed culture of two different blood types neonates cord blood hematopoietic progenitors with single sample and found little difference between them in number and growth characteristics. This may provide a theoretical basis for clinical transplantation of mixed human cord progenitor cells.

Blood Group Antigens↗

PubMed1994

Ligand (T3) dependent and independent effects of thyroid hormone receptors upon human TRH gene transcription in neuroblastoma cells.

Thyrotropin releasing hormone (TRH) gene is regulated negatively at the transcriptional level by thyroid hormone (T3) in rat anterior hypothalamus. The actions of T3 upon other target genes are known to be mediated through the thyroid hormone receptors (TR), TR alpha and TR beta. To explore whether the inhibitory regulation of human (h) TRH gene transcription by T3 is TR isoform specific and whether TRH gene transcription can be modulated as well by unliganded TR isoforms, transient gene expression studies have been carried out using hTRH-luciferase (TRH-Luc) chimeric constructs and TR expression constructs, co-transfected into a human neuroblastoma cell line (HTB-11). Data herein demonstrate T3-dependent inhibitory regulation of the hTRH gene promoter by TR-T3 complexes. Moreover, significant inhibition (39%-60%) could be achieved by T3 bound to either hTR alpha 1, hTR beta 1, or rTR beta 1, beta 2 and was comparable quantitatively, indicating an absence of TR isoform specificity for T3 inhibition. Conversely, basal promoter activity of the hTRH gene could be activated significantly by unliganded hTR alpha 1, beta 1, rTR beta 1, and beta 2 (150% to 334%), but not by hTR alpha 2. Thus, TRs appear to exert opposite effects on hTRH gene transcription, depending on the presence or absence of ligand (T3). These dual effects of TR suggest that the addition of the T3 ligand effects conformational changes that can abrogate the initiation of transcription.

Animals↗

PubMed1994

Identification of thyrotropin-releasing hormone receptor in the rat testis.

We have recently documented the expression of preprothyrotropin-releasing hormone (TRH) gene in murine, human and rat testis. Moreover, we have localized TRH to rat Leydig cells immunohistochemically, and found that both prepro TRH mRNA and TRH levels are developmentally regulated in the rat testis. To investigate the potential roles of TRH in testicular function, characterization of TRH receptors (TRH-R) in this tissue was undertaken. Recently, a cDNA encoding murine TRH-R has been isolated, making possible cloning of a rat TRH-R cDNA from the anterior pituitary gland. This cDNA was used for detection of TRH-R gene expression in the rat testis by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). TRH receptor assays were also performed with (3H)MeHisTRH as the radioactive ligand. In Northern blot analysis, a single and specific hybridization band, approximately 3.8 kb in size, was identified in whole testis RNA, identical in size with that found in the anterior pituitary gland. The concentration of TRH-R mRNA in the testis was approximately 10% of that in the pituitary. TRH-R mRNA was also detected by RT-PCR in Metrizamide gradient-purified Leydig cells. TRH receptor binding assays revealed the presence of specific, high affinity binding sites with a Kd of 1.6 x 10(-8) M in the testis. Such TRH binding was inhibited by chlordiazepoxide, a specific antagonist of TRH receptor binding. We conclude that TRH may exert local, probably autocrine, actions in the testis via a transmembrane receptor very similar or identical to that in pituitary.

Animals↗

PubMed1994

Genetic analysis of uidA expression in enterohaemorrhagic Escherichia coli serotype O157:H7.

Isolates of enterohaemorrhagic Escherichia coli serotype O157:H7 do not exhibit beta-glucuronidase (GUD) activity; however, they carry nucleotide sequences for the uidA gene that encodes the GUD enzyme. Polymerase chain reaction analysis using uidA-specific primers confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in E. coli O157:H7 isolates. DNA sequencing analysis of the regulatory region and the 5' terminus of the uidA gene of E. coli O157:H7 showed a base substitution in the putative -10 promotor region and at 93 bases downstream from the initiation codon. Neither base alteration, however, appeared to affect uidA allele gene expression in the O157:H7 isolates. Immunoblotting of cell extracts with an anti-GUD antibody showed that E. coli O157:H7 isolates produced an antibody-reactive protein that was homologous in size to E. coli GUD, but no GUD activity was observed in cell-free extracts of these isolates. These results suggest that the antibody-reactive protein produced by E. coli O157:H7 may be an inactive GUD enzyme. Sequencing of the uidA structural gene in O157:H7 showed the presence of 18 additional nucleotide base substitutions, but only six altered the amino acid sequence. Also, there were two frame shift mutations, 18 bases apart, that altered the sequence of six consecutive amino acids. These genetic alterations in the uidA structural gene of O157:H7 may account for the absence of GUD activity in this serotype.

Amino Acid Sequence↗