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Biomedical subjects

P Gu

Publications and source records attributed to P Gu.

At least 19 recordsLinked to original sources
PubMed2002

Observation of the decay K- --> pi(-)mu(+)mu(-) and measurements of the branching ratios for K+/- --> pi(+/-)mu(+)mu(-).

Using data collected with the HyperCP (E871) spectrometer during the 1997 fixed-target run at Fermilab, we report the first observation of the decay K--->pi(-)mu(+)mu(-) and new measurements of the branching ratios for K+/--->pi(+/-)mu(+)mu(-). By combining the branching ratios for the decays K+-->pi(+)mu(+)mu(-) and K--->pi(-)mu(+)mu(-), we measure Gamma(K+/--->pi(+/-)mu(+)mu(-))/Gamma(K+/--->all) = (9.8+/-1.0+/-0.5)x10(-8). The CP asymmetry between the rates of the two decay modes is [Gamma(K+-->pi(+)mu(+)mu(-))-Gamma(K--->pi(-)mu(+)mu(-))]/[Gamma(K+-->pi(+)mu(+)mu(-))+Gamma(K--->pi(-)mu(+)mu(-))] = -0.02+/-0.11+/-0.04.

Journal Article↗
PubMed2001

Mutagenesis of the regulatory domain of phenylalanine hydroxylase.

The regulatory domain of phenylalanine hydroxylase (PAH, EC ) consists of more than 100 amino acids at the N terminus, the removal of which significantly activates the enzyme. To study the regulatory properties controlled by the N terminus, a series of truncations and site-specific mutations were made in this region of rat PAH. These enzymes were expressed highly in Escherichia coli and purified through a pterin-conjugated Sepharose affinity column. The removal of the first 26 amino acids of the N terminus increased the activity by about 20-fold, but removal of the first 15 amino acids increased the activity by only 2-fold. Replacing serine-29 of rat PAH with cysteine from the same site of human PAH increased the activity by more than 4-fold. Mutation of serine to other amino acids with varying side chains: alanine, methionine, leucine, aspartic acid, asparagine, and arginine also resulted in significant activation, indicating a serine-specific inhibitory effect. But these site-specific mutants showed 30--40% lower activity when assayed with 6-methyl-5,6,7,8-tetrahydropterin. Stimulation of hydroxylase activity by preincubation of the enzyme with phenylalanine was inversely proportional to the activation state of all these mutants. Combined with recent crystal structures of PAH [Kobe, B. et al. (1999) Nat. Struct. Biol. 6, 442-448; and Erlandsen, H., Bjorgo, E., Flatmark, T. & Stevens, R. C. (2000) Biochemistry 39, 2208-2217], these data suggest that residues 16-26 have a controlling regulatory effect on the activity by interaction with the dihydroxypropyl side chain of (6R)-5,6,7,8-tetrahydrobiopterin. The serine/cysteine switch explains the difference in regulatory properties between human and rat PAH. The N terminus as a whole is important for maintaining rat PAH in an optimum catalytic conformation.

Amino Acid Sequence↗
PubMed2000

Detection of genetic alterations in mouse lung adenocarcinomas by two-dimensional gel electrophoresis.

DNA from 14 mouse lung adenocarcinomas and 7 normal lungs were examined by 2-dimensional gel electrophoresis (2-DGE) for genetic alterations. 2-DGE profiles from tumor samples were compared with those profiles from normal lung tissues through a computer-assisted color overlay system. Compared to the profiles in normal lung DNA, 6 spots were amplified and 16 spots were partially reduced in their intensity in tumors. Two spots were detectable only in tumor tissues. These altered spots indicate genetic changes in mouse lung tumor development. The identification of these genetic alterations is probably important in understanding mouse lung carcinogenesis.

Adenocarcinoma↗
PubMed2000

[Expression of proteolytic enzymes of metastatic tumor cells and the effects of some influencing factors].

OBJECTIVE: To elucidate the types of proteolytic enzymes the dendritic cell sarcoma (DCS) cell expressed in vitro and the effects of modulating factors. METHODS: The proteolytic spots of DCS cells were examined, with ten different proteinase inhibitors, various antibodies and matrix. RESULTS: Aprotinin, EDTA-Na2 and pepstatin could inhibit the proteolysis of DCS cells respectively. Anti-ubiquitin antibody and anti-DCS antibody (1:200-1:100) showed no obvious inhibitory effects. DCS cells spread out fairly well but with no proteolysis. CONCLUSIONS: Under the conditions of our experiments the DCS cells had serine proteinase, metalloproteinase and aspartate proteinase activities. Antiubiquitin antibodies and anti-DCS antibodies showed no obvious influence on the proteolysis of DCS cells. DCS cells displayed different proteolysis status on different matrix.

Aspartic Acid Endopeptidases↗
PubMed1999

[The effects of 6A8 cDNA transfection on biological behavior of tumor cells].

OBJECTIVE: To investigate the effects of antisense 6A8 cDNA on the malignant behavior of tumor. METHODS: Highly metastatic clonal variant CNE-2L2 from nasopharygeal carcinoma cell line(CNE-2Z) was transfected with antisense 6A8 cDNA or mock plasmid. Their adhesion, locomotion, proteolytic ability, in vitro invasion, in vivo growth and metastasis were comparatively studied with the wild CNE-2L2 cells. RESULTS: Antisense 6A8 cDNA transfected CNE-2L2 showed less adhesion to FN and LN, weakened locomotive ability and proteolytic ability, retarded in vivo growth and much less metastasis. CONCLUSION: Transfection of antisense 6A8 cDNA obviously decreased the malignant behavior of tumor cells.

Cell Adhesion↗
PubMed1998

Function-structure studies and identification of three enzyme domains involved in the catalytic activity in rat hepatic squalene synthase.

Rat hepatic squalene synthase (RSS, EC 2.5.1.21) contains three conserved sections, A, B, and C, that were proposed to be involved in catalysis (McKenzie, T. L., Jiang, G., Straubhaar, J. R., Conrad, D., and Shechter, I. (1992) J. Biol. Chem. 267, 21368-21374). Here we use the high expression vector pTrxRSS and site-directed mutagenesis to determine the specific residues in these sections that are essential for the two reactions catalyzed by RSS. Section C mutants F288Y, F288L, F286Y, F286W, F286L, Q293N, and Q283E accumulate presqualene diphosphate (PSPP) from trans-farnesyl diphosphate (FPP) with reduced production of squalene. F288L, which retains approximately 50% first step activity, displays only residual activity (0.2%) in the production of squalene from either FPP or PSPP. Substitution of either Phe288 or Phe286 with charged residues completely abolishes the enzyme activity. Thus, F288W, F288D, F288R, F286D, and F286R cannot produce squalene from either FPP or PSPP. All single residue mutants in Section A, except Tyr171, retain most of the RSS activity, with no detectable accumulation of PSPP in an assay mixture complete with NADPH. Y171F, Y171S, and Y171W are all inactive. Section B, which binds the diphosphate moieties of the allylic diphosphate subtrates, contains four negatively charged residues: Glu222, Glu226, Asp219, and Asp223. The two Glu residues can be replaced with neutral or with positively charged residues without signficantly affecting enzyme activity. However, replacement of either Asp residues with Asn eliminates all but a residual level of activity, and substitution with Glu abolishes all activity. These results indicate that 1) Section C, in particular Phe288, may be involved in the second step of catalysis, 2) Tyr171 of Section A is essential for catalysis, most likely for the first reaction, 3) the two Asp residues in Section B are essential for the activity and most likely bind the substrate via magnesium salt bridges. Based on these results, a mechanism for the first reaction is proposed.

Amino Acid Sequence↗
PubMed1998

[Peripheral artery blood flow measurements using pulsed Doppler and electromagnetic flowmetry in dogs: a comparative study].

To investigate the accuracy of blood flow parameters in peripheral artery by Doppler ultrasound, the intermal diameter(D), peak velocity(Vp), mean velocity (Vm) and flow volume (Q) of dog's femoral artery were measured by using either a pulsed Doppler (PD) system or electromagnetic flowmeters(EMFM). The results showed that the D measured by B-mode ultrasound values significantly smaller than that by callipers but it correlated very well with the D by callipers. When the values of Vp, Vm and Q measured by pulsed Doppler and EMFM were compared, the Vp had the most highly significant correlation between the data obtained by the two techniques (r: 0.91, 0.79, 0.83 respectively). When Vp, Vm and Q were measured by angle correction of PD, the correlations between the data determined by the two techniques were still significant(r: 0.88, 0.82, 0.78).

Animals↗
PubMed1997

Characterization of a novel microsomal fatty acid synthetase (FAS) compared to a cytosolic FAS in the housefly, Musca domestica.

A novel membrane-bound fatty acid synthetase (FAS) associated with the microsomal fraction from the housefly, Musca domestica, was solubilized and purified to homogeneity. The microsomal FAS was solubilized by 0.75 M KCl in phosphate buffer and was purified to homogeneity by the sequential use of ammonium sulfate precipitation followed by Sepharose CL-6B, DEAE Sephacel and Red Agarose (dye ligand affinity) chromatography. The specific activity of the microsomal FAS was increased 1,440-fold to 6,522 U/mg during purification. The cytosolic FAS from the housefly was also purified by similar methods and the specific activity increased 183-fold to 7,533 U/mg. The relative molecular mass of the microsomal and cytosolic FAS are 419 +/- 22 kDa and 405 +/- 18 kDa, respectively, for the dimers as determined by gel permeation chromatography. The microsomal and the cytosolic FAS yield different tryptic digestion maps and have slightly different amino acid compositions, which demonstrate structural differences between the two FASs. In addition, there are differences between the two FASs in their kinetic characteristics and their ability to incorporate methylmalonylCoA into the growing fatty acyl chain.

Acyl Coenzyme A↗
PubMed1996

Detection of genomic alterations in human cervical cancer by two-dimensional gel electrophoresis.

Two-dimensional gel electrophoresis was used to comprehensively scan the whole genome of 6 cervical intraepithelial neoplasia (CIN) lesions, 7 cervical squamous cell carcinomas, 1 cervical adenosquamous cell carcinoma, and 2 cervical adenocarcinomas for multiple genetic alterations, such as DNA amplification, chromosome deletion, loss of heterozygosity, and chromosome translocation, as compared with the paired normal tissues. DNA spot analysis of the genomic 2-dimensional gels was performed by a computer color overlay system and by spot recognition software allowing for objective spot comparison and quantitation. Nine spots were found to be amplified in the cervical carcinomas while two amplified spots were detected in the CIN III lesions. Fourteen DNA spots were either reduced in their intensity or absent in cervical carcinomas as compared to their normal paired tissues. Reduction of intensity in 6 spots was observed in the 5 CIN III lesions. These genetic alterations may represent changes in cancer genes that are associated with human cervical carcinogenesis. Further characterization of these alterations may be significant to the understanding of cervical tumorigenesis and to the development of biomarkers for clinical trials in cancer chemoprevention.

Adenocarcinoma↗
PubMed1995

Regulation of enzymatic activity involved in sex pheromone production in the housefly, Musca domestica.

Ovarian produced ecdysteroids regulate sex pheromone production in the female housefly, inducing the synthesis of (Z)-9-tricosene (Z9-23:Hy), cis-9,10-epoxytricosane, (Z)-14-tricosen-10-one and methylalkanes. Experiments were performed to gain a detailed understanding of the processes affected by 20-hydroxyecdysone (20-HE) that result in sex pheromone production as the female becomes reproductively mature. A novel microsomal fatty acid synthetase (FAS) is present in the epidermal tissue and plays a role in producing the methyl-branched fatty acid precursors to the methylalkanes. This FAS is released from the microsomes in the presence of 3 M KCl. A major enzyme activity influenced by 20-HE is the fatty acyl-CoA elongation system. A shift in the chain length specificity of the products of the elongation system causes the change in the chain lengths of the alkenes produced to switch from C27 and longer in the previtellogenic female to C23 in the mature female. Data is presented indicating that it is the condensation activity of the elongation system that is affected. Z9-23:Hy arises from a 24 carbon acyl group which is reduced to an aldehyde, and then converted to the hydrocarbon. Data is presented demonstrating that it is the fatty acyl-CoA derivative and not the free fatty acid that is the substrate. There does not appear to be a chain length specificity which regulates the conversion of fatty acyl-CoAs to hydrocarbons as both 24 and 28 carbon fatty acyl-CoAs are converted to hydrocarbon by both males and females of all ages.

Acyl Coenzyme A↗