PubMed Health⌕ Search

Biomedical subjects

X Ding

Publications and source records attributed to X Ding.

At least 163 records · Page 9Linked to original sources

Pulses with fixed magnitude and variable phase response profiles.

The Shinnar-Le Roux (SLR) method of pulse envelope design provides for the generation of nearly arbitrary magnitude response profiles with such great efficiency that the pulse envelopes may often be calculated at the time of sequence initiation. A significant limitation of the method is that it provides only limited control of the phase of the response profile. In the current manuscript it is demonstrated that the phase of the response profile can be modulated without affecting the magnitude profile by replacement of some of the roots of the SLR polynomials with one over their complex conjugate. This method allows interactive tailoring of the phase profile to the user's needs. Although the method does not allow for the optimization of arbitrary phase profiles, a variety of pulses, which are of general utility, have been generated. Some of these pulses and their response profiles are presented.

Humans↗

Metabolism of aflatoxin B1 by rabbit and rat nasal mucosa microsomes and purified cytochrome P450, including isoforms 2A10 and 2A11.

The nasal mucosa of some mammalian species are susceptible to the toxicity of aflatoxin B1 (AFB1), a potent hepatocarcinogen, but little is known about the nasal enzymes involved in the metabolic activation of AFB1 or the metabolites produced. In the present study, the metabolism of AFB1 was studied with nasal microsomes from rats and rabbits and with several purified isozymes of rabbit P450 in a reconstituted enzyme system. The rates of AFB1-N7-guanine DNA adduct formation with rabbit and rat nasal microsomes are over 3- and 10-fold higher, respectively, than with liver microsomes from the same species. On the other hand, the rates of formation of AFM1 (9a-hydroxy-AFB1) and AFQ1 (3-hydroxy-AFB1) products known to be less toxic, are lower with nasal than with liver microsomes. Of particular interest, nasal microsomes produce high levels of six unidentified polar metabolites that are not formed by microsomes from liver or several other tissues. These same products are also generated by P450 NMa purified from rabbit nasal microsomes in a reconstituted system, but not by five other isozymes of cytochrome P450 (1A2, 2B4, 2E1, 2G1, 3A6) that are known to be present in nasal microsomes. AFB1-DNA adducts are formed by P450 NMa at a rate 3-fold higher than that by nasal microsomes. The DNA adducts are formed at much slower rates by P450s 2G1, 2B4, and 1A2, and adducts are not formed at measurable rates by P450s 2E1 and 3A6. Moreover, AFB1-DNA adduct formation is also catalyzed by cDNA-derived, heterologously expressed P450s 2A10 and 2A11, both of which are known to be present in the purified P450 NMa preparation. The Km and Vmax values of the two isozymes for DNA adduct formation are comparable to those for nasal microsomes. Furthermore, the formation of AFB1-DNA adducts by nasal microsomes is decreased by nicotine, a known inhibitor of P450 NMa. These data indicate that members of the P450 2A gene subfamily play an important role in the metabolic activation of AFB1 in rabbit and rat nasal mucosa and suggest a molecular basis for assessing the health risk associated with inhalation exposure to this procarcinogen in humans.

Aflatoxin B1↗

Zonula occludens toxin modulates tight junctions through protein kinase C-dependent actin reorganization, in vitro.

The intracellular signaling involved in the mechanism of action of zonula occludens toxin (ZOT) was studied using several in vitro and ex vivo models. ZOT showed a selective effect among various cell lines tested, suggesting that it may interact with a specific receptor, whose surface expression on various cells differs. When tested in IEC6 cell monolayers, ZOT-containing supernatants induced a redistribution of the F-actin cytoskeleton. Similar results were obtained with rabbit ileal mucosa, where the reorganization of F-actin paralleled the increase in tissue permeability. In endothelial cells, the cytoskeletal rearrangement involved a decrease of the soluble G-actin pool (-27%) and a reciprocal increase in the filamentous F-actin pool (+22%). This actin polymerization was time- and dose-dependent, and was reversible. Pretreatment with a specific protein kinase C inhibitor, CGP41251, completely abolished the ZOT effects on both tissue permeability and actin polymerization. In IEC6 cells ZOT induced a peak increment of the PKC-alpha isoform after 3 min incubation. Taken together, these results suggest that ZOT activates a complex intracellular cascade of events that regulate tight junction permeability, probably mimicking the effect of physiologic modulator(s) of epithelial barrier function.

Actins↗

[Full thickness lamellar keratoplasty with viscodelamination of cornea for treatment of bullous keratopathy].

Intrastromal injection of viscoelastic (2% methylcellulose or healon) will facilitate the separation of deep stroma during dissection of recipient stroma. Full thickness lamellar keratoplasty was performed on a smooth graft bed which approached Descemet's membrane. 21 cases with bullous keratopathy were treated and followed up for six months, post-operatively. The symptoms were controlled in all cases and 13 cases were relieved from blindness with corrected visual acuities over 0.05. The safety, efficacy and reoperability of full thickness lamellar keratoplasty with viscodelamination of cornea for treatment of bullous keratopathy make it a promising technique.

Adolescent↗

Structure-function analysis of CYP2A10 and CYP2A11, P450 cytochromes that differ in only eight amino acids but have strikingly different activities toward testosterone and coumarin.

Cytochrome P450 NMa, which was first identified in this laboratory in rabbit nasal microsomes, is now known to represent two distinct gene products, P450s 2A10 and 2A11. In the present study, chimeric and site-directed mutants of 2A11 were constructed to determine which of the eight different amino acid residues are responsible for the much greater activity of 2A10 toward coumarin and testosterone. Mutation of Arg62 and Asp63 of 2A11 to the corresponding residues in 2A10, or mutation of Thr120 to Ser, as found in 2A10, did not change the activities. However, mutation of Arg62, Asp63, Gln104, Ala117, and Thr120 of 2A11 to the corresponding residues in 2A10 resulted in a protein that is as active as 2A10 in coumarin hydroxylation and approximately half as active as 2A10 in androstenedione formation. Mutation of Arg372 in 2A11 to His, as found in 2A10, resulted in a significant increase in the rate of hydroxylation of testosterone, but not of coumarin. Our findings indicate that the identify of the amino acid at position 104 and/or 117 is important for activity with testosterone and for regioselectivity at the 17 position, as well as for optimal activity with coumarin. In contrast, the identity of the residue at position 372 is important for optimal activity with testosterone but not the regioselectivity at the 17 position and does not influence the activity with coumarin.

Amino Acid Sequence↗

Direct structural observation of an acyl-enzyme intermediate in the hydrolysis of an ester substrate by elastase.

The method of X-ray crystallographic cryoenzymology has been used to determine the crystal structure of a kinetically significant species on the reaction pathway of a crystalline enzyme. The structure of a specific acyl-enzyme intermediate in the elastase-catalyzed hydrolysis of the N-carbobenzoxy-L-alanine p-nitrophenyl ester has been determined and refined against X-ray diffraction data at 2.3-A resolution. The difference Fourier electron density map clearly shows electron density for the trapped acyl-enzyme. The acyl-enzyme was formed at -26 degrees C and was stabilized at -55 degrees C during data collection, taking advantage of the glass transition in protein dynamics that occurs at around -50 degrees C.

Alanine↗

Fragile X induction systems in CVS cultures: effect on cytogenetic, PCR, and genomic Southern Blot DNA analyses of the FMR-1 gene.

Low fragile X frequencies have been commonly observed in chorionic villus sample (CVS) cultures, compared to subsequent analysis in whole blood or products of conception (POC). To investigate possible mechanisms for this effect, CVS cultures from a previously identified fragile X positive male, were restudied and compared to subsequent POC cultures from lung, muscle, skin, and thymus. Cultures were exposed, for the last 24 hours before harvesting, to FUdR, excess thymidine, and a combination of both. For CVS, only those cultures that were exposed to a combination of FUdR and excess thymidine showed positive cytogenetic findings (1/90 or 1.1%), agreeing with our original positive cytogenetic results (2/86 or 2.3%) for cultures exposed to excess thymidine. Fragile X frequencies in the POC tissues from this fetus increased to an average of 14%. PCR analyses showed full mutations (> 200 CGG repeats) in uninduced CVS cultures but induced cultures exhibited apparently smaller sizes in the range of 120-180 repeats. The results showed variability. In one instance, the banding pattern from one of the uninduced cultures was similar to the results where cultures were exposed to a double induction system. When PCR analyses were conducted on induced POC cultures, full mutations were observed in virtually all samples. Southern blot genomic analysis using probe StB12.3 showed an unmethylated full mutation in CVS cultures. Southern blot patterns from cultures of muscle revealed size variations of DNA bands in the premutation range representing unmethylated DNA as well as methylated full mutations. Finally, variations were also observed in lung and skin cultures, compared to CVS and muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Blotting, Southern↗

SPARC (secreted protein acidic and rich in cysteine) regulates endothelial cell shape and barrier function.

SPARC (secreted protein acidic and rich in cysteine) can be selectively expressed by the endothelium in response to certain types of injury and induces rounding in adherent endothelial cells in vitro. To determine whether SPARC might influence endothelial permeability, we studied the effect of exogenous SPARC on the movement of 14C-labeled bovine serum albumin across postconfluent bovine pulmonary artery endothelial cells. SPARC increased (P < 0.02) transendothelial albumin flux in a dose-dependent manner at concentrations > or = 0.5 microgram/ml. At a fixed dose (15 micrograms/ml), exposure times > or = 1 h augmented (P < 0.005) albumin flux by 1.3- to 3.6-fold; this increase was blocked by anti-SPARC antibodies but not by inhibition of protein synthesis. Barrier dysfunction was not associated with loss of cell viability. Monolayers exposed to SPARC exhibited a rounded morphology and intercellular gaps. Prior stabilization of F-actin with phallicidin protected against the changes in barrier function (P = 0.0001) that were otherwise induced by SPARC. Bovine aortic and retinal microvascular endothelia also responded to SPARC. We propose that SPARC regulates endothelial barrier function through F-actin-dependent changes in cell shape, coincident with the appearance of intercellular gaps, that provide a paracellular pathway for extravasation of macromolecules.

Actins↗

Steroid metabolism by rabbit olfactory-specific P450 2G1.

Cytochrome P450 2G1 (2G1), which is uniquely expressed in the olfactory mucosa in mammals, may have important physiological functions. In the present study, we have examined the catalytic activity of rabbit 2G1 toward a number of steroid sex hormones, including androstenedione, estradiol, progesterone, testosterone, and 5 alpha-dihydrotestosterone; the purified cytochrome is active toward all of these compounds in a reconstituted enzyme system with turnover numbers of 1.84, 0.34, 1.46, 1.04, and 0.84, respectively, at a substrate concentration of 5 microM. In the presence of cytochrome b5, the turnover numbers are 1.58, 0.66, 1.66, 2.74, and 1.34, respectively. Estradiol is converted to the 2-hydroxy compound (major product) and 4-hydroxy compound (minor product) by 2G1, and progesterone is converted to the 16 alpha-hydroxy derivative as well as the corresponding keto compound as a secondary product. The same products are formed in olfactory microsomal suspensions as major metabolites of progesterone, and the reactions are inhibited strongly by anti-2G1 IgG. In a reconstituted system, 2G1 has an apparent Km of 2.0 microM and a Vmax of 1.8 nmol/min/nmol P450 for the formation of the 16 alpha-hydroxyprogesterone. Of particular interest, 2G1-catalyzed progesterone metabolism is effectively inhibited by the boar pheromones, 5 alpha-androst-16-en-3-one and 5 alpha-androst-16-en-3 alpha-ol, and to a lesser extent by a variety of odorant compounds as well as by known P450 inhibitors, including ketoconazole and alpha-naphthoflavone. The broad substrate specificity and relatively high catalytic efficiency of 2G1 in sex steroid metabolism suggest a role for this unique P450 isozyme in the maintenance of steroid hormone homeostasis in the olfactory mucosa.

Androstenedione↗

Bioconcentration of cadmium in water hyacinth (Eichhornia crassipes) in relation to thiol group content.

To study the bioconcentration of cadmium in water hyacinth, the plants were exposed to water containing 2 microg Cd2+/ml for extended periods of time. Three strains from several exposures during a 30-day period were sampled for the analyses of cadmium and thiol group. The data showed that the plant concentrates cadmium mainly in the roots and that the cadmium uptake is proportional to the increase of the thiol group content. The latter suggests the possibility of using the thiol group content to assess the bioconcentration of heavy metal ions in water hyacinth and as a general parameter for monitoring the heavy metal pollution of water. A simple two-compartmental model was used to simulate the kinetics of cadmium uptake. The calculated bioconcentration factor matches the one derived directly from experimental data, indicating the adequacy of the model.

Journal Article↗

Analogous inhibitors of elastase do not always bind analogously.

It has been assumed that the structure of a single inhibitor complex is sufficient to define the available subsites of an enzyme that has a unique binding site and a uniquely defined mode for ligand binding--the specificity for these subsites can thus be probed by kinetic experiments. Elastase is an enzyme for which these traditional assumptions, which underlie such structural and kinetic studies, do not hold. Three new crystal structures of elastase complexed to chemically similar inhibitors with similar binding affinities reveal a diversity of binding modes as well as two new subsites on elastase. The existence of multiple binding sites and different binding modes for such similar inhibitors indicates that researchers must proceed with caution when using kinetics to map out protein subsites.

Anilides↗

Lipopolysaccharide (LPS)-binding protein and soluble CD14 function as accessory molecules for LPS-induced changes in endothelial barrier function, in vitro.

Bacterial LPS induces endothelial cell (EC) injury both in vivo and in vitro. We studied the effect of Escherichia coli 0111:B4 LPS on movement of 14C-BSA across bovine pulmonary artery EC monolayers. In the presence of serum, a 6-h LPS exposure augmented (P < 0.001) transendothelial 14C-BSA flux compared with the media control at concentrations > or = 0.5 ng/ml, and LPS (10 ng/ml) exposures of > or = 2-h increased (P < 0.005) the flux. In the absence of serum, LPS concentrations of up to 10 micrograms/ml failed to increase 14C-BSA flux at 6 h. The addition of 10% serum increased EC sensitivity to the LPS stimulus by > 10,000-fold. LPS (10 ng/ml, 6 h) failed to increase 14C-BSA flux at serum concentrations < 0.5%, and maximum LPS-induced increments could be generated in the presence of > or = 2.5%. LPS-binding protein (LBP) and soluble CD14 (sCD14) could each satisfy this serum requirement; either anti-LBP or anti-CD14 antibody each totally blocked (P < 0.00005) the LPS-induced changes in endothelial barrier function. LPS-LBP had a more rapid onset than did LPS-sCD14. The LPS effect in the presence of both LBP and sCD14 exceeded the effect in the presence of either protein alone. These data suggest that LBP and sCD14 each independently functions as an accessory molecule for LPS presentation to the non-CD14-bearing endothelial surface. However, in the presence of serum both molecules are required.

Acute-Phase Proteins↗

Predictors of outcome after anterior temporal lobectomy: positron emission tomography.

We assessed the relationship between temporal lobe metabolism measured quantitatively and qualitatively with PET using [18F]-fluorodeoxyglucose (FDG) and postoperative seizure frequency after anterior temporal lobectomy. Forty-three patients with refractory partial epilepsy had anterior temporal lobectomy and preoperative assessment with PET-FDG. Qualitative PET analysis was performed visually by two blinded observers, and quantitative PET analysis was performed using an anatomic template for six control and six temporal lobe subregions, deriving an asymmetry index for each region. Seizure outcome was assessed 1 year after surgery; patients were classified as being seizure-free or as having persistent seizures. Qualitative data were analyzed using Fisher's exact test and the t test, and quantitative data were analyzed using a repeated-measures ANOVA. Thirty-two patients (74%) were seizure-free at follow-up, and 11 had persistent seizures, although most improved. Twenty-nine of 35 patients (83%) with restricted temporal lobe hypometabolism by visual analysis were seizure-free, compared with three of eight patients (37.5%) with normal scans or multilobar hypometabolism. Quantitative analysis revealed that an asymmetry of mesial temporal lobe glucose consumption (uncal region) correlated with improved surgical outcome (p < 0.02). We developed a logistic regression model to predict individual outcome based on the asymmetry in uncal metabolism. Lateral temporal metabolism did not correlate with outcome. We conclude that both visual PET analysis and quantitative PET analysis predict outcome after temporal lobectomy, although quantitative measures offer more precise information.

Adolescent↗

Sequential three-dimensional time-of-flight MR angiography of the carotid arteries: value of variable excitation and postprocessing in reducing venetian blind artifact.

OBJECTIVE: Multiple overlapping three-dimensional (3D) time-of-flight carotid MR angiography potentially combines many of the desirable features of two-dimensional (2D) and single-volume 3D MR angiographic imaging techniques. Yet the maximum-intensity-projection images from such acquisitions are often degraded by artifact due to nonuniform signal intensity of contiguous imaging volumes and inadequate, yet arduous, postprocessing. The former has been termed venetian blind artifact. To date, the severity of the artifact has been minimized by the use of very thin slabs with a large percentage of overlap. However, the artifact typically is still appreciable, and the required acquisition and postprocessing times are increased. The purpose of this study was to examine the value of technical modifications of both the multislab acquisition and postprocessing procedures to reduce this artifact on images of healthy volunteers. SUBJECTS AND METHODS: Spatially variable RF pulses along the direction of flow were applied as excitation pulses in the multislab time-of-flight MR angiographic acquisitions to compensate for the nonuniform blood signal intensity caused by spin saturation. An automatic postprocessing technique was used to optimally combine the image data in overlapping slices by selecting the higher-intensity pixel of the two on a pixel-by-pixel basis. Ratios of the standard deviation of signal intensity to the mean signal intensity were computed as a function of RF profile and postprocessing method along the long axes of the arteries to measure the uniformity of the signal intensity of the blood. The spatially variable and sinc RF pulse acquisitions, combined with automatic and conventional manual postprocessing, were compared. RESULTS: Compared with the sinc pulse acquisition, the MR angiograms acquired with spatially variable excitation pulses improved the signal uniformity of the arteries with thicker volumes and less overlap, thereby reducing the acquisition time by 25% for similar spatial coverage. When used with the automatic postprocessing technique, the severity of the venetian blind artifact on maximum-intensity-projection images was minimized and the postprocessing time was reduced by roughly a factor of 5. CONCLUSION: The combined use of spatially variable excitation pulses and an automatic postprocessing technique can improve the uniformity of the signal from blood across the slab and allow thicker slabs to be acquired with less overlap. Data acquisition and postprocessing times can be reduced significantly. This work suggests it may be possible to easily produce overlapping 3D MR angiograms that should be superior to conventional 2D and 3D studies.

Adult↗

[Studies on the induction of L-phenylalanine ammonia lyase(PAL) in Rhodotorula glutinis and transformation of phenylalanine from trans-cinnamic acid].

The induction of PAL in Rhodotorula glutinis and transformation of L-phenylalanine from trans-cinnamic acid were studied. The optimum medium for PAL induction was composed of (g/L) 10.0 yeast extract, 10.0 peptone, 5.0 NaCl, 0.5 KH2 PO4, 0.5 phenylalanine 1.0 (NH4)2SO4 and 5.0 glucose, pH 6.0-6.5. The cultivation temperature was 30.0 degrees C. In the process of transformation, the effect of [NH4]+ on initial velocity was in accordance with Michaelis-Menten rate expression in which Km and Vmax were 16.85 mol/L and 5.96 g.L-1.h-1 for ammonia and the optimum pH was 10.0. Substrate activation and inhibition were observed at low and high concentration of cinnamic acid. The yield of phenylalanine from cinnamic acid reached more than 60.0%.

Cinnamates↗

Rapid fragile X carrier screening and prenatal diagnosis using a nonradioactive PCR test.

OBJECTIVE: To develop a rapid, nonradioactive test using the polymerase chain reaction (PCR) capable of detecting full fragile X mutations, premutations, and resolving normal alleles and to apply this to prenatal diagnosis and carrier screening of pregnant women at risk for fragile X carrier status. DESIGN: Prenatal and blood sample PCR analysis with confirmation by direct Southern blotting and cytogenetic techniques. SETTING: Samples sent to a DNA diagnostic research laboratory at a tertiary referral center. PARTICIPANTS: Pregnant women with a family history of undiagnosed mental retardation or known fragile X syndrome and controls. RESULTS: A rapid, nonradioactive PCR screening protocol for the fragile X mental retardation-1 gene for both normal and mutant alleles was developed. Analysis of 570 control X chromosomes showed a modal number of 30 CGG repeats (range, 12 to 52 repeats) and a calculated heterozygosity of approximately 80%. No excess of homozygosity was found, indicating the test was accurate for normal allele resolution. In addition, 150 unrelated pregnant women were screened. Within known fragile X families, five of 20 pregnant women were diagnosed as carriers. Two new fragile X families were diagnosed among relatives of 130 females with family histories of undiagnosed mental retardation, although no carriers were identified. Prenatal PCR testing of 28 carriers accurately detected nine fetuses with full mutations. CONCLUSIONS: This rapid, nonradioactive PCR protocol allows accurate resolution of normal alleles as well as simultaneous detection of carrier alleles and full mutations. With this approach, efficient screening of pregnant women at risk for fragile X carrier status, subsequent genetic counseling of identified carriers, and reliable prenatal diagnosis can be offered.

Base Sequence↗

Isolation and heterologous expression of cloned cDNAs for two rabbit nasal microsomal proteins, CYP2A10 and CYP2A11, that are related to nasal microsomal cytochrome P450 form a.

Nasal microsomal P450 form a (NMa), a major cytochrome P450 isozyme in rabbit olfactory and respiratory nasal mucosa with high activity toward a variety of odorants and environmental toxicants, was previously purified to electrophoretic homogeneity from rabbit nasal microsomes. In the present study, a cDNA library constructed from poly(A)+ RNA from rabbit respiratory nasal mucosa was screened with antibodies to P450 NMa, and five immunopositive clones were isolated and characterized. Sequence analysis indicated that the clones encode two highly similar P450s that contain 494 amino acid residues, with the first 20 corresponding to P450 NMa, and differ from each other in only 8 residues scattered throughout the polypeptide chains. On the basis of structural homology the two proteins are designated as CYP2A10 and CYP2A11 and are the first members of the P450 2A subfamily to be identified in nasal tissue. Genomic blot analysis indicated that 2A10 and 2A11 are apparently not allelic variants. Both genes are expressed in liver and lung as well as in nasal tissues, as judged by RNA blot analysis, but the relative levels of the two mRNAs differ. Both enzymes were partially purified after expression of the cDNAs in Escherichia coli and shown to catalyze the oxygenation of a variety of substrates, including ethanol and procarcinogens such as N-nitrosodiethylamine and phenacetin. P450 2A10 is generally more active than P450 2A11 and strikingly so in the conversion of testosterone to androstenedione.

Amino Acid Sequence↗

Bacterial lipopolysaccharide induces actin reorganization, intercellular gap formation, and endothelial barrier dysfunction in pulmonary vascular endothelial cells: concurrent F-actin depolymerization and new actin synthesis.

Bacterial lipopolysaccharide (LPS) influences pulmonary vascular endothelial barrier function in vitro. We studied whether LPS regulates endothelial barrier function through actin reorganization. Postconfluent bovine pulmonary artery endothelial cell monolayers were exposed to Escherichia coli 0111:B4 LPS 10 ng/ml or media for up to 6 h and evaluated for: 1) transendothelial 14C-albumin flux, 2) F-actin organization with fluorescence microscopy, 3) F-actin quantitation by spectrofluorometry, and 4) monomeric G-actin levels by the DNAse 1 inhibition assay. LPS induced increments in 14C-albumin flux (P < 0.001) and intercellular gap formation at > or = 2-6 h. During this same time period the endothelial F-actin pool was not significantly changed compared to simultaneous media controls. Mean (+/- SE) G-actin (micrograms/mg total protein) was significantly (P < 0.002) increased compared to simultaneous media controls at 2, 4, and 6 h but not at 0.5 or 1 h. Prior F-actin stabilization with phallicidin protected against the LPS-induced increments in G-actin (P = 0.040) as well as changes in barrier function (P < 0.0001). Prior protein synthesis inhibition unmasked an LPS-induced decrement in F-actin (P = 0.0044), blunted the G-actin increment (P = 0.010), and increased LPS-induced changes in endothelial barrier function (P < 0.0001). Therefore, LPS induces pulmonary vascular endothelial F-actin depolymerization, intercellular gap formation, and barrier dysfunction. Over the same time period, LPS increased total actin (P < 0.0001) and new actin synthesis (P = 0.0063) which may be a compensatory endothelial cell response to LPS-induced F-actin depolymerization.

Actins↗