Biomedical subjects

Y Fu

Publications and source records attributed to Y Fu.

At least 109 records · Page 6Linked to original sources

PubMed1998

Treatment planning optimization by quasi-Newton and simulated annealing methods for gamma unit treatment system.

The gamma unit is used to irradiate a target within the brain. During such a treatment many parameters, including the number of shots, the coordinates, the collimator size and the weight associated with each shot, affect the amount of dose delivered to the target volume and to the surrounding normal tissues. Hence it is not easy to determine an appropriate set of these parameters by a trial and error method. For this reason, we present here an optimization method to determine mathematically those parameters. This method is composed of two steps: firstly, a quasi-Newton method is used to deal with the continuous variables such as position and weight of shots; the result obtained at the end of this step then serves as the initial configuration for the next step, in which a simulated annealing method is applied to optimize all the aforementioned parameters. Application of the proposed methods to two examples shows that our optimization algorithm runs in a satisfactory way.

Algorithms↗

PubMed1998

Mutagenesis of the glucose-1-phosphate-binding site of potato tuber ADP-glucose pyrophosphorylase

Lysine (Lys)-195 in the homotetrameric ADP-glucose pyrophosphorylase (ADPGlc PPase) from Escherichia coli was shown previously to be involved in the binding of the substrate glucose-1-phosphate (Glc-1-P). This residue is highly conserved in the ADPGlc PPase family. Site-directed mutagenesis was used to investigate the function of this conserved Lys residue in the large and small subunits of the heterotetrameric potato (Solanum tuberosum) tuber enzyme. The apparent affinity for Glc-1-P of the wild-type enzyme decreased 135- to 550-fold by changing Lys-198 of the small subunit to arginine, alanine, or glutamic acid, suggesting that both the charge and the size of this residue influence Glc-1-P binding. These mutations had little effect on the kinetic constants for the other substrates (ATP and Mg2+ or ADP-Glc and inorganic phosphate), activator (3-phosphoglycerate), inhibitor (inorganic phosphate), or on the thermal stability. Mutagenesis of the corresponding Lys (Lys-213) in the large subunit had no effect on the apparent affinity for Glc-1-P by substitution with arginine, alanine, or glutamic acid. A double mutant, SK198RLK213R, was also obtained that had a 100-fold reduction of the apparent affinity for Glc-1-P. The data indicate that Lys-198 in the small subunit is directly involved in the binding of Glc-1-P, whereas they appear to exclude a direct role of Lys-213 in the large subunit in the interaction with this substrate.

Journal Article↗

PubMed1998

ADP-Glucose pyrophosphorylase from potato tubers. Site-directed mutagenesis studies of the regulatory sites.

Several lysines (Lys) were determined to be involved in the regulation of the ADP-glucose (Glc) pyrophosphorylase from spinach leaf and the cyanobacterium Anabaena sp. PCC 7120 (K. Ball, J. Preiss [1994] J Biol Chem 269: 24706-24711; Y. Charng, A.A. Iglesias, J. Preiss [1994] J Biol Chem 269: 24107-24113). Site-directed mutagenesis was used to investigate the relative roles of the conserved Lys in the heterotetrameric enzyme from potato (Solanum tuberosum L.) tubers. Mutations to alanine of Lys-404 and Lys-441 on the small subunit decreased the apparent affinity for the activator, 3-phosphoglycerate, by 3090- and 54-fold, respectively. The apparent affinity for the inhibitor, phosphate, decreased greater than 400-fold. Mutation of Lys-441 to glutamic acid showed even larger effects. When Lys-417 and Lys-455 on the large subunit were mutated to alanine, the phosphate inhibition was not altered and the apparent affinity for the activator decreased only 9- and 3-fold, respectively. Mutations of these residues to glutamic acid only decreased the affinity for the activator 12- and 5-fold, respectively. No significant changes were observed on other kinetic constants for the substrates ADP-Glc, pyrophosphate, and Mg2+. These data indicate that Lys-404 and Lys-441 on the small subunit are more important for the regulation of the ADP-Glc pyrophosphorylase than their homologous residues in the large subunit.

Amino Acid Sequence↗

PubMed1998

Expression of the Candida albicans gene ALS1 in Saccharomyces cerevisiae induces adherence to endothelial and epithelial cells.

To identify genes encoding adhesins that mediate the binding of Candida albicans to endothelial cells, a genomic library from this organism was constructed and used to transform Saccharomyces cerevisiae. These transformed organisms were screened for adherence to endothelial cells, and a highly adherent clone was identified. The adherence of this clone to endothelial cells was over 100-fold greater than that of control S. cerevisiae transformed with the empty plasmid. This clone also exhibited enhanced adherence to epithelial cells. The C. albicans gene contained within this clone was found to be ALS1. These results indicate that ALS1 may encode a candidal adhesin.

Adhesiveness↗

PubMed1998

Cloning and characterization of CAD1/AAF1, a gene from Candida albicans that induces adherence to endothelial cells after expression in Saccharomyces cerevisiae.

Adherence to the endothelial cell lining of the vasculature is probably a critical step in the egress of Candida albicans from the intravascular compartment. To identify potential adhesins that mediate the attachment of this organism to endothelial cells, a genomic library from C. albicans was used to transform a nonadherent strain of Saccharomyces cerevisiae. The population of transformed yeasts was enriched for highly adherent clones by repeated passages over endothelial cells. One clone which exhibited a fivefold increase in endothelial cell adherence, compared with S. cerevisiae transformed with vector alone, was identified. This organism also flocculated. The candidal DNA fragment within this adherent/flocculent organism was found to contain a single 1.8-kb open reading frame, which was designated CAD1. It was found to be identical to AAF1. The predicted protein encoded by CAD1/AAF1 contained features suggestive of a regulatory factor. Consistent with this finding, immunoelectron microscopy revealed that CAD1/AAF1 localized to the cytoplasm and nucleus but not the cell wall or plasma membrane of the transformed yeasts. Because yeasts transformed with CAD1/AAF1 both flocculated and exhibited increased endothelial cell adherence, the relationship between adherence and flocculation was examined. S. cerevisiae expressing either of two flocculation phenotypes, Flo1 or NewFlo, adhered to endothelial cells as avidly as did yeasts expressing CAD1/AAF1. Inhibition studies revealed that the flocculation phenotype induced by CAD1/AAF1 was similar to Flo1. Thus, CAD1/AAF1 probably encodes a regulatory protein that stimulates endothelial cell adherence in S. cerevisiae by inducing a flocculation phenotype. Whether CAD1/AAF1 contributes to the adherence of C. albicans to endothelial cells remains to be determined.

Adhesiveness↗

PubMed1998

Identification of a specific chaperone for SptP, a substrate of the centisome 63 type III secretion system of Salmonella typhimurium.

Salmonella typhimurium uses of a type III protein secretion system encoded at centisome 63 of its chromosome to deliver effector molecule into the host cell. These proteins stimulate host cell responses such as reorganization of the actin cytoskeleton and activation of transcription factors. One of these effector proteins is SptP, a tyrosine phosphatase that causes disruption of the host cell actin cytoskeleton. A characteristic feature of many substrates of type III secretion systems is their association with specific cytoplasmic chaperones which appears to be required for secretion and/or translocation of these proteins into the host cell. We report here the identification of SicP, a 13-kDa acidic polypeptide that is encoded immediately upstream of sptP. A loss-of-function mutation in sicP resulted in drastically reduced levels of SptP but did not affect sptP expression, indicating that SicP exerts its effect posttranscriptionally. Pulse-chase experiments demonstrated that the loss of SicP leads to increased degradation of SptP. In addition, we show that SicP binds to SptP directly and that the binding site is located between residues 15 and 100 of the tyrosine phosphatase. Taken together, these results indicate that SicP acts as a specific chaperone for SptP.

Actins↗

PubMed1998

Endothelial nitric oxide synthase increases in left atria of dogs with pacing-induced heart failure.

In congestive heart failure (CHF) the alterations in cardiac NO synthase (NOS) isoforms activity and expression are incompletely documented and the chamber specificity of these changes is unknown. We studied plasma nitrate-nitrite (NO-x), atrial, and ventricular NOS activities and protein expression (Western blot and densitometric analysis) in nonpaced control dogs and in dogs paced for 2 or 21 days into CHF. Plasma NO-x rose significantly after 7 and 21 days of pacing, whereas creatinine levels remained unchanged. In control dogs Ca2+-dependent NOS activity in left atria was double that of right or left ventricular activity. In paced animals the activity increased only in the atria after 21 but not 2 days of pacing. Levels of endothelial NOS (eNOS) protein were enhanced in the left atria but not ventricles after 21 days of pacing because of a greater quantity of the 150-kDa but not the 135-kDa eNOS. Ca2+-independent NOS activity was undetectable in any cardiac tissue. The specific upregulation of eNOS in the left atria suggests that NO production may be enhanced to counterbalance hypertrophy that develops during pacing-induced CHF.

Animals↗

PubMed1998

Vertebrate tinman homologues XNkx2-3 and XNkx2-5 are required for heart formation in a functionally redundant manner.

Tinman is a Drosophila homeodomain protein that is required for formation of both visceral and cardiac mesoderm, including formation of the dorsal vessel, a heart-like organ. Although several vertebrate tinman homologues have been characterized, their requirement in earliest stages of heart formation has been an open question, perhaps complicated by potential functional redundancy of tinman homologues. We have utilized a novel approach to investigate functional redundancy within a gene family, by coinjecting DNA encoding dominantly acting repressor derivatives specific for each family member into developing Xenopus embryos. Our results provide the first evidence that vertebrate tinman homologues are required for earliest stages of heart formation, and that they are required in a functionally redundant manner. Coinjection of dominant repressor constructs for both XNkx2-3 and XNkx2-5 is synergistic, resulting in a much higher frequency of mutant phenotypes than that obtained with injection of either dominant repressor construct alone. Rescue of mutant phenotypes can be effected by coinjection of either wild-type tinman homologue. The most extreme mutant phenotype is a complete absence of expression of XNkx2-5 in cardiogenic mesoderm, an absence of markers of differentiated myocardium, and absence of morphologically distinguishable heart on the EnNkxHD-injected side of the embryo. This phenotype represents the most severe cardiac phenotype of any vertebrate mutant yet described, and underscores the importance of the tinman family for heart development. These results provide the first in vivo evidence that XNkx2-3 and XNkx2-5 are required as transcriptional activators for the earliest stages of heart formation. Furthermore, our results suggest an intriguing mechanism by which functional redundancy operates within a gene family during development. Our experiments have been performed utilizing a recently developed transgenic strategy, and attest to the efficacy of this strategy for enabling transgene expression in limited cell populations within the developing Xenopus embryo.

Amino Acid Sequence↗

PubMed1998

[A study on dose-effect of suppression to gap junctional intercellular communication function by 50-Hz magnetic fields].

OBJECTIVE: To study the relationship between dose of exposure to 50-Hz magnetic field (MF) and its effects of suppression to gap junctional intercellular communication (GJIC) function. METHODS: Lucifer yellow, a kind of fluorescent dye, was led into single CHL cell by iontophoretic injection. Number of dye-coupled cells (DCC) five minutes after injection was used as an indicator for GJIC function. The effects of different intensities and irradiation of MF on GJIC in CHL cells were studied. RESULTS: Suppression to GJIC by 50-Hz MF depended on its flux intensity. Irradiation with lower flux intensity (0.05, 0.2 and 0.4 mT) of the MF for 24 hours could not inhibit GJIC, but that with higher intensity (0.8 and 1.6 mT) could. No "amplitude window" effect was appeared by irradiation with MF at intensities of zero to 1.6 mT. Exposure to MF at a fixed intensity of 0.8 mT for five minutes had no effects, but that for one hour was more than enough to inhibit GJIC, with the most apparent effects for 24-hour exposure. In addition, it was showed that suppression to GJIC was caused by direct action of 50-Hz MF rather then induced electrical field. CONCLUSION: MF of 50-Hz had suppression to GJIC with a dose-effect relationship.

Animals↗

PubMed1998

Erythrocyte and plasma Ca2+, Mg2+ and cell membrane adenosine triphosphatase activity in patients with essential hypertension.

OBJECTIVE: To assess the relationships between plasma and intracellular Ca2+, Mg2+ and blood cell membrane adenosine triphosphatase (ATPase) activity in normotensive and hypertensive subjects. METHODS: Plasma and intracellular Ca2+, Mg2+ were measured with atomic absorption spectrophotometry, and red blood cell membrane Na(+)-K+ ATPase and Ca(2+)-ATPase activities were determined with colorimetric method in 55 patients with essential hypertension and 32 normotensive controls. RESULTS: The results showed that the hypertensive group consistently demonstrated a significant decreased activity of ATPase studied, with significantly lower plasma Ca2+ and higher cytosolic Ca2+ levels when compared with those in normotensive group (P < 0.01 or P < 0.05, respectively). No significant differences were found in either plasma Mg2+ or intracellular Mg2+ level between the two groups. CONCLUSIONS: This study suggests that patients with essential hypertension have widespread depression of cell membrane Na(+)-K(+)-ATPase and Ca(2+)-ATPase activities with plasma Ca2+ depletion and cytosolic Ca2+ overload, which may reflect an underlying membrane abnormality in essential hypertension. The cellular abnormalities may be related to the defective transport mechanisms that in turn may be aggravated by plasma Ca2+ depletion.

Adult↗

PubMed1998

Detection of the E7 transform gene of human papilloma virus type 16 in human oral squamous cell carcinoma.

OBJECTIVE: To determine, with the use of polymerase chain reaction, the prevalence of human papillomavirus (HPV) 16 in 30 patients with primary oral squamous cell carcinoma (OSCC) and 30 healthy control patients. MATERIALS AND METHODS: DNA was extracted from freshly frozen tumor tissues of 30 patients with primary oral squamous cell carcinoma and from the oral mucosa of 30 controls. A pair of specific primers of the E7 early gene of HPV 16 were designed. PCR products were run by 1.5% agarose gel and the results of electrophoresis were photographed. RESULTS: HPV 16 was detected in 36.7% (11/30) of oral squamous cell carcinoma patients and 11.1% (4/30) of controls. CONCLUSIONS: HPV 16 has a significant association with oral squamous cell carcinoma. However, the role HPV 16 plays in the tumorigenesis of oral cancer and its clinical significance remain to be investigated.

Adult↗

PubMed1998

[Circulating ICAM-1 in sera of renal allograft recipients in monitoring of acute rejection].

OBJECTIVE: To study the significance of ICAM-1 in sera of renal allograft recipients in mornitoring of acute rejection. METHODS: The levels of circulating intercellular adhesion molecule-1 (cICAM-1) in sera of 87 renal transplantation recipients, including 29 patients with acute rejection and 8 patients with cyclosprine nephrotoxicity were measured by sandwich-ELISA. RESULTS: The levels of cICAM-1 were elevated transiently after transplantation. They decreased and reached normal levels within 2 weeks. In the acute rejection episode, the levels of cICAM-1 were significantly higher than those of patients with stable graft function and cyclosporine nephrotoxicity (P < 0.001). The increase of cICAM-1 was seen 1-3 days early than the diagnosis was made. On successful antirejection therapy, the levels of cICAM-1 decreased to normal, but in patients with steroid-resistant rejection, it did not decrease. CONCLUSION: The cICAM-1 in sera can be used to estimate the graft function, and it is also useful in prediction and differential diagnosis of acute rejection and prognosis.

Adult↗

PubMed1998

[The pressure of amniotic cavity during pregnancy and treatment of unruptured oligohydramnios with antepartum amnioinfusion].

OBJECTIVES: To measure the pressure of the amniotic cavity during pregnancy and to study the effects of antepartum amnioinfusion on oligohydramnios with intact membranes. METHOD: We used an improved three-way switch apparatus to measure 110 women during pregnancy, and treated 97 oligohydramnios with antepartum amnioinfusion or intravenous infusion as controls. RESULTS: The pressure of the amniotic cavity was 1.69 +/- 0.23 kPa, which was close to the normal value in pregnancy. In oligohydramnios, it was 1.1 +/- 0.3 kPa, and in polyhydramnios 3.2 +/- 0.3 kPa(P < 0.01-0.001). When 300 ml fluid was amnioinfused, the amniotic fluid index (AFI) increased by 5.0 +/- 1.8 cm, the pressure of amniotic cavity increased by 0.18 +/- 0.2/kPa and the incidence of vaginal delivery was 87.9%, whereas that of the control group was 12.1%. There were no fetal distress, asphyxia neonatorum, amnionitis or ruptured membrane. CONCLUSIONS: The pressure of the amniotic cavity during normal pregnancy was stable, but changed with AFI. Antepartum amnioinfusion for unruptured oligohydramnios resulted in a marked increase in incidence of vaginal delivery, a decreased ratio of cesarean section and a reduction of perinatal morbidity. These differences are highly significant.

Adult↗

PubMed1998

[Microsurgical technique in the treatment of lumbar disc prolapse].

From 1983 to 1996, 354 cases of the lumbar intervertebral disc prolapse were treated by the microsurgical technique in our hospital. Among them, 330 cases were the simple prolapse of the lumbar intervertebral disc, the other 24 cases were complicated with the lateral recess stenosis. The application of the microsurgical technique has the advantage of its precision, shorter operative time, little surgical trauma, less bleeding, and quicker recovery. So the patients can usually get up and move around at an earlier time. Ninety-five percent of the patients acquired good results. It is suggested that the microsurgical technique is a relatively good operative method in the treatment of the lumbar intervertebral disc prolapse and the lateral recess stenosis.

Adolescent↗

PubMed1998

[Rapid determination of essential fatty acids of edible oils by conversion to their hydroxamic acids].

A simple and rapid HPLC method for the determination of essential fatty acids of edible oils was established. Oil samples were converted to their hydroxamic acids in a single step and analyzed without prior separation and purification. The chromatographic conditions were: Shim-pack CLC ODS, 5 microns, 150 mm x 6.0 mm i.d. column, 40 degrees C; MeOH: 0.02 mol/L NaH2PO4(pH 3.0) (81:19, V/V) as eluent and UV-213 nm detector. The linear range was 0.05-0.6 g/L, recovery was 96.93% and RSD was 1.80%(n = 4). The relative standard deviations for intra-day and inter-day assays were 1.24% and 1.62% respectively (n = 6). For 18:3, 18:2, 18:1, the difference between the derivatization yields from triglycerides and their methyl esters was almost one fold. That was confirmed by our recovery and determination results. The calibration curves for the oil samples should not be obtained from the derivatization of their methyl ester standards.

Chromatography, High Pressure Liquid↗

PubMed1998

Nicotine-induced norepinephrine release in the rat amygdala and hippocampus is mediated through brainstem nicotinic cholinergic receptors.

Previous studies have shown that nicotine stimulates norepinephrine (NE) release in the rat hypothalamic paraventricular nucleus, which in turn activates the hypothalamo-pituitary-adrenal axis. In the present study, nicotine induced NE release in the amygdala (AMYG) and the hippocampus (HP) of the same rat in vivo. Nicotine (0.065-0.135 mg/kg i.v. at a rate of 0.09 mg/kg/60 sec) dose-dependently increased NE release at both sites with similar potencies. To determine whether the site of action of nicotine is in the brainstem, which contains the noradrenergic cell bodies projecting to AMYG and HP, nicotinic cholinergic receptor (NAchR) antagonists were injected into the cerebral aqueduct before i.v. nicotine. Use of the following antagonists enabled partial characterization of the NAchRs mediating NE secretion: mecamylamine (Mec), dihydro-beta-erythroidine (DH beta E), methyllycaconitine (MLA) and alpha-bungarotoxin (alpha-BTX). Mec inhibited 80% of NE release in AMYG and 87% in HP (IC50 = 6 nmol for both regions). DH beta E blocked 62% of NE release in AMYG (IC50 = 8 nmol) and 63% in HP (IC50 = 15 nmol). Similar to DH beta E, MLA inhibited 60% of NE release in AMYG and 66% in HP (IC50 = 5 nmol for both regions). In contrast, alpha-BTX had no effect on NE release in either region. These results indicate that brainstem NAchRs accessible from the fourth ventricle mediate nicotine-stimulated NE secretion in AMYG and HP. Taken together with prior investigations showing the brainstem expression of mRNAs encoding NAchR subtypes and the selectivity of antagonists for NAchR subtypes, the present studies suggest that brainstem alpha-3 subunits may be involved.

Amygdala↗

PubMed1998

[The types and distributive features of gene mutation on beta-thalassemia in three districts in northwest China].

OBJECTIVE: To sum up the results of gene analyses on beta-Thalassemia in three districts in northwest China and discuss the types and distributive features of the mutation. METHODS: Polymerase china reaction in combination with dot-blot hybridization of allele-specific oligonucleutide probes(PCR-ASO). RESULTS: In the gene analysis carried out in 85 probands with beta-Thalassemia, twelve types of gene mutation were identified from five nationalities. CD8(-AA), CDs8-9(+G), CDs27-28(+C) heterozygote and [-28(A-->uG). CD17(A-->uT)/N] double heterozygote were firstly assayed in these districts besides the common types in Chinese, of which, [-28(A-->uG). CD17(A-->uT)/N] as double gene mutation in the same chromosome is rare. CONCLUSION: There are significant differences in the distribution and nationality features of gene mutation types for beta-Thalassemia in the three districts, so the three districts as a whole is an area with special distributive features of the disease.

Adolescent↗

PubMed1997

Characterization and chromosomal localization of PTPRO, a novel receptor protein tyrosine phosphatase, expressed in hematopoietic stem cells.

Hematopoietic stem cells (HSCs) support blood cells throughout life by utilizing their self-renewing and multilineage differentiating capabilities. Hematopoietic growth factors mediate their effects on stem cells by the tyrosine phosphorylation of proteins. Regulation of tyrosine phosphorylation is partially mediated by protein tyrosine phosphatases (PTPases). A possible mechanism by which hematopoietic stem cells maintain their self-renewing capacity and undifferentiated state is by controlling the balanced and opposing actions of protein tyrosine kinases (PTKs), receptors for growth factors, and PTPases. We have characterized the expression of PTPases in 5-fluorouracil (5-FU)-treated murine bone marrow cells, which represent a very primitive population of progenitors enriched for reconstituting stem cells, by using a consensus polymerase chain reaction (PCR) method. Several PTPases were expressed abundantly in the 5-FU-treated bone marrow stem cells. A novel PTP, termed protein tyrosine phosphatase receptor omicron (PTPRO), which is related to the homotypically adhering kappa, mu and PCP-2 receptor-type tyrosine phosphatases, was identified and characterized. We have cloned the murine and full-length human PTPRO cDNAs which share 89% homology, indicating that PTPRO is highly conserved between these species. The human PTPRO cDNA clone encodes a polypeptide of 1439 amino acids (aa) and has a calculated molecular mass of approximately 162 kDa. PTPRO consists of an extracellular segment containing a MAM domain, an immunoglobulin (Ig) domain, four fibronectin-type III (FN-III) repeats, a transmembrane segment, and two tandem intracellular PTP domains. The human PTPRO gene was assigned to human chromosome 1p35-pter using Southern blot analyses of genomic DNAs from rodent/human somatic hybrid cell lines containing human chromosome 1 or the p35-pter region of the chromosome. The mouse Ptpro gene was mapped to chromosome 4, closely linked to D4Mit16 and Elp1 (elliptocytosis-1), by using genomic DNAs from a (C57BL/6J x Mus spretus)F1 x Mus spretus backcross. In fetal tissues, PTPRO expression was observed in the brain and lung, whereas lower levels were observed in the kidney. In adult tissues, PTPRO was less restricted and was observed in the lung, heart, skeletal muscle, prostate, testis, and in various areas of the brain, indicating that PTPRO expression is developmentally regulated. Expression of PTPRO was also observed in human CD34+ bone marrow cells and 5-FU-treated murine primitive stem cells. These results suggest a potential role for PTPRO in stem cell adhesion and in mediating homophilic cell-cell interactions in other cell types.

Amino Acid Sequence↗