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Biomedical subjects

F Yu

Publications and source records attributed to F Yu.

At least 127 records · Page 7Linked to original sources
PubMed1995

Galanin and somatostatin inhibition of neurokinin A and B induced airway mucus secretion in the rat.

Neurokinin A and B are present in neurons situated in lung and NK-1 receptors have been described on tracheal submucosal gland cells. In the present study we compared the ability of substance P (SP), neurokinin A (NKA) and neurokinin B (NKB) to stimulate airway mucus secretion. Furthermore, we characterized the interaction of NKA and NKB with galanin and somatostatin. The rank order of the tachykinins to stimulate airway mucus secretion was SP > NKA > NKB suggesting that NK-1 receptors mediate these effects(EC50:SP: 50 nmol/l, NKA: 200 nmol/l, NKB: 400 nmol/l). Galanin and somatostatin were equally potent to inhibit NK-A and NK-B stimulated airway mucus release. These results suggest that NK-A and NK-B are potent stimulators of airway macromolecule secretion. Galanin and somatostatin potently inhibit these actions of the tachykinins. Therefore, airway mucus secretion is controlled by a complex network of several different mediators.

Animals↗
PubMed1995

Galanin and somatostatin inhibition of substance P-induced airway mucus secretion in the rat.

Substance P is present in several neurons innervating the lung. Tachykinin receptors are expressed on submucosal gland cells. Substance P is known to be a potent stimulator of airway mucus secretion. In the present study we characterized the effects of galanin and somatostatin on basal and substance P-induced mucus secretion. The stimulatory effect of substance P was concentration-dependent (100 pmol/l: 112%, 1 nmol/l: 120%, 10 nmol/l: 153%, 100 nmol/l: 223%, 1 mumol/l: 275%, 10 mumol/l: 172%) and was inhibited by galanin and somatostatin (1 mumol/l substance P: 277%; 1 mumol/l substance P + 1 mumol/l somatostatin: 190%, p < 0.01; 1 mumol/l substance P + 1 mumol/l galanin: 206%, p < 0.05). In the presence of lower concentrations of substance P 1 mumol/l somatostatin and 1 mumol/l galanin did not modify mucus secretion. Lower concentrations of galanin and somatostatin did not significantly change mucus secretion stimulated by 1 mumol/l substance P. Both, galanin and somatostatin at 1 mumol/l left basal airway mucus secretion unaltered. These data suggest that mucus secretion into airways is regulated by a complex network of peptidergic stimulators and inhibitors including substance P, somatostatin and galanin.

Animals↗
PubMed1995

Molecular cloning and characterization of the cDNA coding for the biotin-containing subunit of the chloroplastic acetyl-coenzyme A carboxylase.

We report the molecular cloning and sequence of the cDNA coding for the biotin-containing subunit of the chloroplastic acetylcoenzyme A (CoA) carboxylase (ACCase) of Arabidopsis thaliana (CAC1). The 3' end of the CAC1 sequence, coding for a peptide of 94 amino acids, which includes a putative biotinylation motif, was expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. The resulting GST-CAC1 fusion protein was biotinylated in vivo, indicating that CAC1 codes for a biotin-containing protein. Antibodies generated to the GST-CAC1 protein reacted solely with the 38-kD biotin-containing polypeptide of Arabidopsis. Furthermore, these antibodies inhibited ACCase activity in extracts from Arabidopsis leaves. The deduced amino acid sequence of CAC1 has an apparent N-terminal chloroplast-targeting transit peptide. The CAC1 protein is coded by a single Arabidopsis gene, and its mRNA accumulates to the highest levels in organs that are undergoing rapid growth. The amino acid sequence of the CAC1 protein is most similar to the biotin carboxyl-carrier protein component of eubacterial ACCases. These characterizations identify CAC1 as the biotin-containing subunit of the plastidic, heteromeric ACCase of Arabidopsis. The results support the ancient origin of the two structurally distinct ACCases of plants.

Acetyl-CoA Carboxylase↗
PubMed1995

[A double-blind parallel group study of beclomethasone dipropoinate nasal spray of two different sources in the treatment of allergic rhinitis].

In order to compare the clinical efficacy and safety of beclomethasone dipropionate nasal spray (BDNS) manufactured in Chongqing Glaxo Limited (Group A) and Glaxo UK (Group B), a randomised double-blind parallel group study was performed. A total of 204 patients with seasonal or perennial allergic rhinitis were recruited into the study for a period of 2 weeks. Overall efficacy (excellent/good) was 88% for Group A (excellent 55%) and 90% for Group B (excellent 51%) respectively. Side effects were similar for both groups where dry nose was the most common complaint (4.9% for both groups). However, there was no statistical difference (P > 0.05) between the two groups either in efficacy or side effects. Pollen count was monitored throughout the study period. Pollen season was divided into three periods, i.e. the beginning, the peak and the end. There was no statistical difference (P > 0.05) between the two groups in any period.

Administration, Intranasal↗
PubMed1994

TNF-beta produced by human T lymphotropic virus type I-infected cells influences the proliferation of human endothelial cells and fibroblasts.

Human T lymphotropic virus type I (HTLV-I) is linked to adult T cell leukemia as well as to HTLV-I-associated myelopathy/tropical spastic paraparesis. In this report, we studied the effects of HTLV-I-infected cell supernatants on HUVEC, fibroblasts, and glioma cells. The HTLV-I-infected cell supernatants (HUT102 and MT-2) strongly inhibited the proliferation of HUVEC, although they enhanced the proliferation of the fibroblasts. Regarding the glioma cells, only the MT-2 supernatant showed weak inhibitory effects on the proliferation. However, the HTLV-I-uninfected cell supernatants showed no effects on these target cells. The biologic activities of both HUT102 and MT-2 supernatants were found to be dose dependent and were reduced by heat treatment at 100 degrees C for 5 min, but not at 56 degrees C for 30 min. These activities were not dependent on the concentrations of HTLV-I viral particles and were only minimally affected by the presence of anti-HTLV-I Abs. A bioassay of various cytokines revealed that the activity of TNF was much higher in the HUT102 and MT-2 supernatants than in the HTLV-I-uninfected cell supernatants (MOLT-4, Jurkat, and K-562). rTNF-alpha and rTNF-beta also showed strong inhibitory effects on HUVEC as well as on the enhancement of the fibroblast growth. With the use of Sephadex G-100 column chromatography, we obtained the highest activities from the 60- through 70-kDa fractions of the HUT102 supernatant and some activities from the 20- through 30-kDa fractions. The biologic activities of both the whole HUT102 supernatant and its active fractions were completely blocked by anti-TNF-beta mAb, although they were not blocked by anti-TNF-alpha mAb. In a Western blot assay, the 25- and 27-kDa bands of TNF-beta were shown clearly in the HUT102 supernatant, although no TNF-alpha bands appeared. These findings suggest that TNF-beta is present in either its oligomeric or monomeric form in the HTLV-I-infected cell supernatants and is also mainly responsible for the supernatants' effects on HUVECs and fibroblasts.

Cell Division↗
PubMed1994

A 17S multiprotein form of murine cell DNA polymerase mediates polyomavirus DNA replication in vitro.

We have identified and purified a multiprotein form of DNA polymerase from the murine mammary carcinoma cell line (FM3A) using a series of centrifugation, polyethylene glycol precipitation, and ion-exchange chromatography steps. Proteins and enzymatic activities associated with this mouse cell multiprotein form of DNA polymerase include the DNA polymerases alpha and delta, DNA primase, proliferating cell nuclear antigen (PCNA), DNA ligase I, DNA helicase, and DNA topoisomerases I and II. The sedimentation coefficient of the multiprotein form of DNA polymerase is 17S, as determined by sucrose density gradient analysis. The integrity of the murine cell multiprotein form of DNA polymerase is maintained after treatment with detergents, salt, RNase, DNase, and after chromatography on DE52-cellulose, suggesting that the association of the proteins with one another is independent of nonspecific interaction with other cellular macromolecular components. Most importantly, we have demonstrated that this complex of proteins is fully competent to replicate polyomavirus DNA in vitro. This result implies that all of the cellular activities required for large T-antigen dependent in vitro polyomavirus DNA synthesis are present within the isolated 17S multiprotein form of the mouse cell DNA replication activities. A model is proposed to represent the mammalian Multiprotein DNA Replication Complex (MRC) based on the fractionation and chromatographic profiles of the individual proteins found to co-purify with the complex.

Animals↗
PubMed1994

Translocation (8;21) in oligoblastic leukemia: is this a true myelodysplastic syndrome?

An 8;21 translocation with trisomy 4 is described in a 36-year-old Chinese woman who presented with an oligoblastic leukemia with myelodysplastic (MDS) features. Progression to acute myeloblastic leukemia (AML) occurred 3 months after presentation. She died of septicemia without remission. Through a review of the data in 10 cases of oligoblastic leukemia with t(8;21) in the literature, we make the following comments. (i) Oligoblastic leukemia with t(8;21) represents 2-3% of cases with this karyotype. (ii) Such cases behave in a similar manner to de novo AML. (iii) The presence of features of MDS has no affect on the behaviour of the disease. (iv) Such cases should be treated without delay with intensive chemotherapy.

Adult↗
PubMed1994

Characterization of a novel enantioselective halohydrin hydrogen-halide-lyase.

Enzymes Ia and Ib of Corynebacterium sp. strain N-1074 exhibit halohydrin hydrogen-halide-lyase (H-lyase) activity, catalyzing the interconversion of halohydrins to epoxides and hydrogen halide. H-lyase B produced in a recombinant Escherichia coli strain carrying one of the enzyme genes of Corynebacterium sp. strain N-1074 was purified and characterized. The purified enzyme catalyzed the transformation of prochiral 1,3-dichloro-2-propanol (DCP) into R-rich epichlorohydrin (ECH). The apparent Km values for DCP, ECH, and chloride were calculated to be 1.03, 5.00, and 4.00 mM, respectively. Maximum activity for the conversion of DCP to ECH was found at pH 8.0 to 9.0, and that for the reverse reaction was found at about pH 5.0. H-lyase B seemed to be identical to enzyme Ib of Corynebacterium sp. strain N-1074 from the comparison of the properties of each. The properties of H-lyase B and H-lyase A, which had been previously purified from another recombinant carrying its gene from Corynebacterium sp. strain N-1074, were also compared.

Amino Acid Sequence↗
PubMed1994

Cloning of two halohydrin hydrogen-halide-lyase genes of Corynebacterium sp. strain N-1074 and structural comparison of the genes and gene products.

We cloned the genes from Corynebacterium sp. strain N-1074, for two halohydrin hydrogen-halide-lyase isoenzyme (hheA and hheB), which are involved in the transformation of alpha, beta-halohydrin into the corresponding epoxide and the reverse reaction. The nucleotide sequences of 1057 base pairs including the hheA gene and of 1130 base pairs including the hheB gene were analyzed. The predicted amino acid sequence of the hheA gene product consisted of 244 residues, and the calculated molecular weight was 26,465. The hheB gene had two sets of potential ribosome binding site and a possible initiation codon. The predicted amino acid sequences of the gene products consisted of 235 and 227 residues, and the calculated molecular weights were 26,179 and 25,236, respectively. Site-directed mutagenesis experiments strongly suggested dual translational initiation in the hheB gene. Comparison of the predicted amino acid sequences for the hheA and hheB genes found significant homology only in the carboxyl terminal region. An analysis of the upstream regions of the both hhe genes suggested the presence of putative epoxide hydrolase genes, which might be involved in further degradation of epoxide compounds.

Amino Acid Sequence↗
PubMed1994

National food contamination monitoring programmes-levels of mercury, lead and cadmium in Chinese foods.

Six hundred and thirty-four food samples of six kinds were collected from Beijing, Shanghai, Jiangsu, Sichuan, Jilin and Guangdong areas, China and Hg, Pb, Cd contents were determined. The results showed that their levels in the Chinese foods were low; their levels in meat, egg, milk and fish were generally below the national hygienic standard. The average daily dietary intake of Hg, Pb, Cd were 7.25 micrograms, 103.77 micrograms and 30.72 micrograms respectively and they were all less than the ADI established by WHO.

Adult↗
PubMed1994

[Effect of prednisolon, vincristine and mitoxantrone on glucocorticoid receptor in HL-60 cell line].

The effects of prednisolon, vincristine and Mitoxantrone on glucocorticoid receptor (GCR) content and affinity were examined in HL-60 cell line with a whole-cell assay in vitro. The fall in GCR number was observed following a short time exposure to prednisolon, vincristine or Mitoxantrone alone. The significant decrease occurred in a dose-dependent (10(-8)-10(-6) mol/L) and time-dependent manner. The treatment of HL-60 cells with 10(-6) mol/L led to a decrease in the affinity of whole cell GCR for [3H]Dex, but vincristine had no that effect. Synergia in GCR content was found during the treatment of HL-60 cells with prednisolon plus Mitoxantrone or prednisolon plus vincristine. The significance of these effects is discussed.

Dose-Response Relationship, Drug↗
PubMed1993

Differential modulation of protein kinase C isoforms in erythroleukemia during induced differentiation.

Induction of erythroid differentiation of murine erythroleukemia cells (MELC) by exposure to hexamethylene bisacetamide (HMBA) involves the modulation of protein kinase C (PKC) activity. Using immuno- and Northern blot techniques, we have demonstrated that MELC express a pattern of PKC isoforms which includes PKC alpha, PKC delta, PKC epsilon, PKC zeta, and PKC eta. We show that MELC resistant to induction by HMBA express significantly less of the nPKC isoform, PKC delta, and slightly less PKC epsilon. Recovery of HMBA sensitivity is associated with reexpression of PKC delta protein. Upon exposure to HMBA, there is a fall in cytosolic PKC delta and PKC epsilon accompanied by a transient increase in membrane-associated forms of these PKC isoforms. HMBA-resistant MELC fail to display this isoform-specific translocation of PKC. Induction of differentiation is accompanied, over the next 24 h of exposure to HMBA, by a progressive fall in cellular PKC activity, associated with a progressive fall in the cellular content of PKC delta, PKC epsilon, and PKC zeta. These studies suggest that PKC delta, and possibly PKC epsilon and PKC zeta as well, play a role in the pathway of HMBA-mediated terminal cell differentiation of MELC.

Animals↗
PubMed1993

The stimulatory effect of amylin on mucus secretion in isolated rat trachea.

A modified Ussing chamber technique was used to characterize the effects of amylin on rat tracheal mucus secretion. The effects of amylin were studied at both the mucosal and the submucosal side of isolated rat trachea. Applied at the mucosal side, amylin had no direct effect on mucus secretion, but significantly decreased the acetylcholine (1 mM)-induced tachyphylaxia when this substance was presented a second time (first stimulation: 490%; second stimulation: 334%; second stimulation plus amylin: 407%). At the submucosal side, amylin significantly stimulated mucus secretion (controls: 100%; 1 microM amylin: 193%; 100 nM amylin: 170%; 10 nM amylin: 127%; 1 nM amylin: 127%). Together with the recent observation of specific amylin receptors in rat lung membranes, these data suggest a physiological role for amylin in the regulation of tracheal mucus secretion.

Acetylcholine↗
PubMed1993

Antihelicase action of DNA-binding anticancer agents: relationship to guanosine-cytidine intercalator binding.

DNA-binding antibiotics such as intercalators, narrow groove binders, and other substances modify duplex DNA, making it an altered substrate for DNA helicases. The intercalators daunorubicin, actinomycin D, echinomycin, and elsamicin, the narrow groove binders distamycin and mithramycin, and the plant toxin teniposide, each representing a different chemical class, block SV40 large T antigen DNA helicase action with IC50 values ranging from 4 x 10(-8) to 2 x 10(-6) M. A partially purified human HeLa cell DNA helicase is also potently blocked by daunorubicin, distamycin, and teniposide. Because eukaryotic cells contain helicases of varying abundance, specificity, and type, this site of action for DNA-binding antibiotics may help explain antibiotic potency and specificity for DNA or RNA inhibition. The antihelicase effect of the antibiotic-double-stranded DNA complex may be central to the anticancer activities of these substances. An additional interesting correlation is the antihelicase action of DNA-intercalating antibiotics and their DNA-binding preference for G-C base pair sites. The G-C base pair binding preference of the intercalating antibiotics may result from evolutionary selection because of the higher G-C binding stability, compared with A-T binding stability. The combination of the higher base pair stability at G-C regions and increased duplex DNA stability induced by intercalating antibiotic yields a total additive stability of the intercalator-G-C base pair complex that resists helicase action.

Animals↗
PubMed1993

Enzymatic and pH modulation of mitomycin C-induced DNA damage in mitomycin C-resistant HCT 116 human colon cancer cells.

The effect of pH and oxygen on DNA alkylation by mitomycin C (MMC) was studied with cell fractions and intact cells. The cell lines used were the HCT 116 human colon cancer cell line and a MMC-resistant subline (HCT 116-R30A) that has 5% of the quinone reductase activity present in the parent cell line. Microsomal fractions of the two cell lines catalyzed MMC-DNA adduct formation only under anaerobic conditions with equal efficiency. However, the pH of the reaction controlled the production of four identified and two unidentified adducts. Soluble fractions from each cell source catalyzed MMC-DNA adduct formation under aerobic and anaerobic conditions similarly. At higher pH, limited DNA adducts were produced by MMC activated by soluble fractions from either cell source. At lower pH, more DNA adducts were obtained with MMC activated by the soluble fraction of HCT 116 cells than with that activated by the soluble fraction of HCT 116-R30A cells. Four of these adducts were identified as N2-(2" beta,7"-diaminomitosene-1" alpha-yl)-2'-deoxyguanylic acid, N2-(2" beta,7"-diaminomitosen-1" beta-yl)-2'-deoxyguanylic acid, N2-(10"-decarbamoyl-2",7"-diaminomitosen-1" alpha-yl)-2'-deoxyguanylic acid, and N2-(2" beta,7"-diamino-10"-deoxyguanyl-N2-yl-mitosen-1" alpha-yl)-2'- deoxyguanylic acid. Acidic intracellular pH enhanced the cytotoxicity of MMC for HCT 116 cells, decreasing the IC50 from 0.3 +/- 0.04 microM to 0.1 +/- 0.03 microM, but pH had limited effect on the cytotoxicity of MMC for HCT 116-R30A cells. When intracellular pH was decreased, interstrand DNA cross-linking by MMC increased to a greater extent in HCT 116 cells than in HCT 116-R30A cells. Only two DNA adducts, each at low intensity, were detected in HCT 116-R30A cells treated at pH 6.0 and 7.6 and in HCT 116 cells treated at pH 7.6. However, six radioactive spots were detected in HCT 116 cells treated at pH 6.0. Three of these adducts were identified. This is the first direct evidence that acidic intracellular pH enhances MMC-DNA adduct formation in tumor cells containing high quinone reductase activity. Results from this study further confirm that pH and not enzyme is the determining factor in the distribution of types of MMC-DNA adducts. This study also indicates that low intracellular pH enhances the activity of quinone reductase in reducing MMC, which is important for aerobic cytotoxicity of MMC against tumor cells with high concentration of quinone reductase.

Biotransformation↗
PubMed1993

Effect of pH on DNA alkylation by enzyme-activated mitomycin C and porfiromycin.

DNA adduct formation by enzyme-activated antibiotics, mitomycin C (MMC) or porfiromycin (PFM), at pH 7.6 or pH 6.0 under anaerobic conditions was analyzed by a 32P-postlabeling method. Antibiotic activation by rat liver NADPH-cytochrome P-450 reductase (EC 1.6.2.4) and bovine milk xanthine oxidase (EC 1.2.3.2) produced similar results. Five 32P-labeled MMC adducts were separated by thin layer chromatography and high performance liquid chromatography from DNA alkylated at either pH. Four of the radioactive spots separated by thin layer chromatography were identified as two monofunctional monoadducts [1" alpha and 1" beta forms of N2-(2" beta,7"-diaminomitosen-1"-yl)-2'-deoxyguanylic acid], one bifunctional monoadduct [N2-(10"-decarbamoyl-2",7"-diaminomitosen-1" alpha-yl)-2'-deoxyguanylic acid], and one cross-linked adduct [N2-(2" beta,7"-diamino-10"-deoxyguanyl-N2-yl-mitosen- 1" alpha-yl)-2'-deoxyguanylic acid]. One minor radioactive spot was not identified. By comparing DNA alkylated at the two pH values, based on equal amounts of 32P radioactivity, similar amounts of cross-links were detected. However, the DNA showed different ratios of the alpha and beta isomers of the monofunctional monoadduct. Furthermore, the DNA alkylated at pH 6.0 showed more bifunctional monoadducts than did the DNA alkylated at pH 7.6. Analysis of alkylated DNA by enzyme-activated PFM showed a similar spectrum of DNA adduct formation. The effect of pH on the distribution of the five PFM-DNA adducts was similar to that observed for the five MMC-DNA adducts. The distribution of adducts in DNA alkylated at the same pH was similar irrespective of which enzyme activated MMC or PFM. The pH of the reaction during DNA and MMC interaction was the determining factor for the quantitative distribution of the adducts. This pH effect may be important for the cytotoxicity of MMC and PFM in tumor cells that have high levels of reductive enzymes with low optimal pH values.

Alkylation↗