Biomedical subjects

K Wang

Publications and source records attributed to K Wang.

At least 271 records · Page 15Linked to original sources

PubMed1998

The reaction of lanthanide ions with n-doxyl stearic acids and its utilization for the ESR study on the permeability of lipid-bilayer of erythrocyte membrane to gadolinium ions.

The reaction of lanthanide ions with n-doxyl stearic acid (nDS) spin labels (n = 5,7,12,16) was investigated by the electron spin resonance technique in aqueous solution. Among the lanthanides, the Gd3+, Tb3+, Tm3+ and Ce3+ ions strongly quenched the ESR signal of spin labels, but the effects of La3+, Eu3+ and Lu3+ are very weak. The quenching effects are featured by: (1) the dependence on the concentration of lanthanide ions; (2) no obvious changes of the ESR line shape in the presence of lanthanide ions; (3) the quenching constant decreases in the order: Gd3+ > Tb3+ > Tm3+ > Ce3+; (4) the quenching effects of lanthanide ions are found to strikingly correlate with their magnetic properties. These findings indicate that the interaction of lanthanide ions with nitroxide oxygen leading to the reduction of ESR signal amplitude is dominated by their magnetic characteristics rather than the coordination effect. By labeling erythrocyte membrane with nDS, n = 5,7,12,16 at different depths, we studied the diffusion of Gd3+ into the lipid-bilayer of erythrocyte membrane by monitoring the reduction processes of the ESR signals of nitroxide spin labels located at different depths of membrane lipid-bilayer after addition of Gd3+. These results revealed that the Gd3+ ions can penetrate into the lipid-bilayer, though the entry rate is slow. It was shown that the Gd3+ ions bind to the membrane and enhance the permeability of extracellular ascorbate into erythrocyte membrane. The transport mechanism of Gd3+ ions through the lipid-bilayer might be involved in the Gd3+ cation-induced pore formation in the surface of membrane.

Ascorbic Acid↗

PubMed1998

Hevin, an antiadhesive extracellular matrix protein, is down-regulated in metastatic prostate adenocarcinoma.

Hevin, a gene closely related to the extracellular matrix protein SPARC, is an acidic cysteine-rich glycoprotein shown to be important for the adhesion and trafficking of cells through the endothelium. Through the use of differential display and differential EST analysis, we identified Hevin as a gene whose transcription is down-regulated in transformed prostate epithelial cell lines and metastatic prostate adenocarcinoma. These results were confirmed by comparing expression levels between normal and neoplastic human prostate tissues using Northern analysis. In situ hybridization with an 35S-labeled antisense riboprobe demonstrated the loss of Hevin expression in metastatic prostate carcinoma. The expression pattern of Hevin in transformed and metastatic epithelium may provide further insights into the complex cell adhesion events involved in the metastatic progression of prostate carcinoma.

Adenocarcinoma↗

PubMed1998

Fluorescence study of the multiple binding equilibria of the galactose repressor.

A fluorescence assay has been developed to study the multiple linked equilibria which function in regulation of the Escherichia coli galactose operon. Fluorescein 5-isothiocyanate was attached to Amino-Modifier C6dT at different positions in an oligonucleotide containing the sequence for the OE site of the galactose operon. These fluorescently labeled oligonucleotides were used to study OEDNA-GalR-d-galactose interactions. The data were analyzed and fit to various models including the classical competitive binding model as well as models involving the formation of a ternary DNA-repressor-inducer complex. Examination of the reduced chi-square of the various fits and comparison of fitted parameters with those obtained in independent experiments were used to distinguish different models. Since the ternary complex is likely to exist under physiological conditions, our results suggest that obligatory dissociation of GalR from DNA may not be required for induction of the gal operon. Rather, induction may involve the formation of a ternary complex of OEDNA with GalR and D-galactose with a different conformation than the GalR-OEDNA binary repressor complex.

Amino Acid Sequence↗

PubMed1998

Effects of thiol protease inhibitors on myoblast fusion and myofibril assembly in vitro.

To investigate the roles of thiol proteases such as cathepsins and calpains in muscle differentiation, we have treated primary cultures of pectoralis muscle with a variety of protease inhibitors and examined the effects these agents have on myoblast fusion and myofibrillogenesis. We have found that a membrane-permeable inhibitor, E64D, has dramatic effects on both events of muscle differentiation. Cells treated with this inhibitor display gross morphological changes, severe delays in myofibril assembly, and reduced ability to fuse in culture. These morphological changes are correlated with a build up of beta1-integrin throughout the cytoplasm. These effects could also be produced using NH4Cl, a lysosomotrophic agent. In addition, we show that two nonpermeable inhibitors (leupeptin and E64) slightly decrease myoblast fusion, but have no effects on the ability of the cells to form mature myofibrils. These results are discussed in terms of their relevance to the inheritable disease of muscular dystrophy and I-cell disease (mucolipodosis II).

Animals↗

PubMed1998

Relation between ventricular fibrillation voltage and probability of defibrillation shocks. Analysis using Hilbert transforms.

We used Hilbert transforms to re-evaluate the previously reported correlation between defibrillation shock outcome and absolute ventricular fibrillation voltage (AVFV). Previously in the literature, single values of AVFV acquired just prior to shock delivery were used to show a correlation between AVFV and shock outcome. In subsequent studies, a modified analysis procedure was used to show a correlation between shock outcome and moving average of AVFV. The use of single values of AVFV makes the AVFV sensitive to the local phase of the electrocardiographic (ECG) waveform, whereas moving averages are weighted by previous values of the ECG. The envelope is independent of the phase of the ECG and is not weighted by previous values of ECG as is the moving average. We explored, therefore, whether the AVFV computed from the envelopes of the ECG showed a stronger correlation with shock outcome than single values and moving averages of the AVFV. We estimated envelopes using the Hilbert transform. Orthogonal ECGs (sagittal, X; transverse, Y; and longitudinal, Z) were recorded from 11 dogs during 10 seconds of electrically induced ventricular fibrillation followed by a defibrillation shock with 50% probability of success. We used transvenous (right ventricular apex to subcutaneous patch) leads to deliver defibrillation shocks. Results from 236 successful and 249 unsuccessful trials showed, in contrast with the previously reported correlation, that moving average of AVFV was not higher for successful trials. In the Z direction, unsuccessful trials had higher voltage than successful trials (P < .05). Comparison of envelope voltages between successful and unsuccessful trials did not show any consistent and statistically significant differences. Although there were some methodological differences between ours and the previously reported studies, they are unlikely to have caused the discrepant observations. Our results suggest, therefore, that the absolute voltage of ECG during ventricular fibrillation is not robustly correlated with shock outcome.

Animals↗

PubMed1998

Optimal harvesting policy for single population with periodic coefficients.

In this paper, we examine the exploitation of single population modeled by time-dependent Logistic equation with periodic coefficients. First, it is shown that the time-dependent periodic Logistic equation has a unique positive periodic solution, which is globally asymptotically stable for positive solutions, and we obtain its explicit representation. Further, we choose the maximum annual-sustainable yield as the management objective, and investigate the optimal harvesting policies for constant harvest and periodic harvest. The optimal harvest effort that maximizes the annual-sustainable yield, the corresponding optimal population level, the corresponding harvesting time-spectrum, and the maximum annual-sustainable yield are determined, and their explicit expressions are obtained in terms of the intrinsic growth rate and the carrying capacity of the considered population. Our interesting and brief results generalize the classical results of Clark for a population described by the autonomous logistic equation in renewable resources management.

Conservation of Natural Resources↗

PubMed1998

Relationship between cellular uptake rate and chemical behavior of diammine/diaminocyclohexane platinum (II) complexes with oxygen-ligating anionic groups.

The uptake kinetics of the platinum (II) complexes of the formula Pt(NH3)2X, Pt(dach)X by human erythrocyte in the plasma isotonic buffer was studied. The results showed that across-membrane transport of all the platinum complexes studied follows a first-order kinetic process. The uptake rate constants decrease with the change of oxygen-ligating anionic group in the sequence: sulfato > selenato > anion of squaric acid > oxalato > anion of demethylcantharic acid > malonato and increase with increasing lipophilicity of carrier group. The relationship between uptake rate and reactivity of these complexes was established. The stereochemistry of dach isomers was shown without effect on the reactivity and the sequence.

Actins↗

PubMed1998

The bidirectional effect of vanadyl ion on the oxygen affinity of human hemoglobin.

The bidirectional effects of vanadyl on the oxygen affinity of hemoglobin depend on the mole ratio of vanadyl to Hb(R). In low R, the vanadyl ion increases Hb's oxygen affinity due to DPG hydrolysis. The lowered oxygen affinity in higher R is mainly due to the reactive-oxygen-species, probably superoxide, induced oxidation of Fe(II)-Hb to Fe(III)-Hb. The conformation change due to vanadyl binding contribute also to the lowered oxygen binding, but is monotonously decreasing with increasing of R values.

Circular Dichroism↗

PubMed1998

Derivatization of 5-fluorouracil with 4-bromomethyl-7-methoxycoumarin for determination by liquid chromatography-mass spectrometry.

A robust new analytical method has been developed for the determination of 5-fluorouracil (5-FU) in human plasma samples using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The method is based on a liquid-liquid extraction procedure, precolumn derivatization, reversed-phase HPLC separation, and detection using atmospheric pressure chemical ionization and selected reaction monitoring. The derivatization agent used was 4-bromomethyl-7-methoxycoumarin. The internal standard for the assay procedure was a stable isotope labeled analog of 5-FU. The lower limit of quantitation was 1.0 ng/mL using 500 microL aliquots of plasma. Sample throughput on the mass spectrometer was approximately 17 samples/h (3.5 min/sample). The method was fully validated. The recovery of 5-FU averaged 76.1%. The accuracy of the assay, assessed from quality control samples, ranged from 99.1% to 104.3% (% theoretical). The overall interassay precision (% RSD) was 2.7%, and the intraassay precision (% RSD) ranged from 1.5% to 3.9%. The derivatized samples were found to be stable under sample analysis conditions and during refrigerator storage. The method was specific for the determination of 5-FU.

Antimetabolites↗

PubMed1998

Synthesis and characterization of bay region halohydrins derived from Benzo[a]pyrene diol epoxide and their role as intermediates in halide-catalyzed cis adduct formation.

The bay region epoxide of benzo[a]pyrene (anti-BPDE) alkylates DNA to form adducts with >98% trans stereochemistry. Halide ions catalyze this reaction; however, this pathway is characterized by the formation of adducts with altered cis stereochemistry. Bay region halohydrins are possible intermediates in these reactions, but are too unstable to be isolated from aqueous solutions. However, we successfully synthesized halohydrins in tetrahydrofuran (THF) by treatment of anti-BPDE with the corresponding lithium halide salt in the presence of acetic acid. Absorbance and CD spectroscopy clearly indicated the formation of chloro-, bromo-, and iodohydrins. The structure and stereochemistry of the chlorohydrin was established by NMR. Chloride addition is exclusively at the benzylic position of the epoxide, and the stereochemistry of the C-9 and -10 positions is trans. The long-wavelength absorbance band in the chloro-, bromo-, and iodohydrin is red-shifted 7, 13, and 22 nm, respectively, relative to the hydrolysis product of anti-BPDE. The ellipticity of the same absorbance band in CD spectra of enantiomerically pure halohydrins is opposite in sign compared to that of the corresponding anti-BPDE enantiomer. The relative stability of these derivatives is chlorohydrin > bromohydrin > iodohydrin. The chloro- and bromohydrins were isolated, but the iodohydrin decomposed during this manipulation. The addition of 500 mM chloride decreased the hydrolysis rate of the chlorohydrin 4-fold in 50% THF/buffer. Direct evidence for the transient formation of the iodohydrin in aqueous buffer/acetone mixtures was obtained by absorbance spectroscopy. At 1 M chloride, bromide, and iodide, alkylation of deoxyadenosine by anti-BPDE in aqueous buffer yields 85, 91, and 92% cis adducts, respectively. In the absence of halide, alkylation of deoxyadenosine in buffer by anti-BPDE, the chlorohydrin, and the bromohydrin yields 32, 65, and 83% cis adducts, respectively.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

PubMed1998

Spatial enhancement of event-related potentials using multiresolution analysis.

Multiresolution analysis is a potentially useful tool to enhance the brain's electrical fields (spatial distributions of event-related potentials (ERP)), and to bring out spatial features which may not be seen in the fields before enhancement. For comparing different images (slices from ERP of different subjects or from the same subject but evoked by different stimuli), we define a measure (surface energy) at each decomposition scale and for different wavelets. The best wavelet and the best level for comparing the given images can be chosen based on this measure. Our experiments show that for very similar images, their difference can be brought out at some scale level. Three preprocessing steps are needed in order to carry out this wavelet analysis. First, a wavelet denoising step is needed to remove noise from the raw ERP. Secondly, a one-to-one mapping is needed to map scalp surface into a square, because the current wavelet analysis theory and algorithm are constructed on regular domains. Finally, a fitting or interpolation step is needed to construct an image on a regular grid in order to apply the fast wavelet transform algorithms.

Brain↗

PubMed1998

A novel frameshift mutation induced by an adenosine insertion in the polycystic kidney disease 2 (PKD2) gene.

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common Mendelian disorders and is genetically heterogeneous. Linkage studies have shown that the majority (approximately 85%) of ADPKD cases are due to mutations in PKD1 on chromosome 16p13.3, while mutations in PKD2 on chromosome 4q21-q23 are thought to account for most of the remaining cases. In this report, we describe the mutation in a large four-generation ADPKD family (TOR-PKD77) which we had mapped to the PKD2 locus by linkage analysis. In this family, we screened for mutations by directly sequencing two nested RT-PCR fragments (PKD2N1 and PKD2N2) that cover approximately 90% of the PKD2 open reading frame. In the affected members, we identified a novel single adenosine insertion (2160InsA) in the PKD2N2 fragment. This mutation occurred in the polyadenosine tract (nt2152-2159) of exon 11 and is predicted to result in a frameshift with premature translation termination of the PKD2 product, polycystin 22, immediately after codon 723. The truncated polycystin 2 is predicted to lack the calcium-binding EF-hand domain and two cytoplasmic domains required for the homodimerization of polycystin 2 with itself and for the heterodimerization of polycystin 2 with polycystin 1.

Adenosine↗

PubMed1998

Treatment of long dissections by use of a single long or multiple short stents: clinical and angiographic follow-up.

BACKGROUND: Recently, long (> or =20 mm) coronary stents were introduced for clinical use. They are intended as an alternative to multiple conventional stents to treat extensive dissections or suboptimal results of long lesions after balloon angioplasty. METHODS: In a total of 113 such consecutive vessels in 107 patients, the flexible Freedom stent was implanted. In 60 of these vessels, because of anatomic constraints, multiple overlapping short (16 mm) stents were implanted. The other 53 vessels were treated with a single long (> or =20 mm) stent. RESULTS: In the single stent group there were four implantation failures (8%) successfully managed by crossover to multiple overlapping short stents. During early follow-up, in-stent thrombosis was not observed, but three patients with a single long stent and two patients with multiple overlapping stents suffered myocardial infarction as a result of long lasting myocardial ischemia during a difficult angioplasty procedure. At 6-month follow-up, > or =50% restenosis was measured in 29% and 35% of the patients with a single long stent and in those with multiple overlapping stents, respectively (not significant). CONCLUSIONS: Compared with the alternative treatment modality (i.e., implantation of multiple short stents), no difference between in-hospital and 6-month outcome was observed. However, implantation of a single long stent, when technically feasible, reduces catheterization time, dye volume for the patient, and radiation exposure for both patient and operator during these embarrassing angioplasty procedures.

Adult↗

PubMed1998

Complete genomic sequence and analysis of the prion protein gene region from three mammalian species.

The prion protein (PrP), first identified in scrapie-infected rodents, is encoded by a single exon of a single-copy chromosomal gene. In addition to the protein-coding exon, PrP genes in mammals contain one or two 5'-noncoding exons. To learn more about the genomic organization of regions surrounding the PrP exons, we sequenced 10(5) bp of DNA from clones containing human, sheep, and mouse PrP genes isolated in cosmids or lambda phage. Our findings are as follows: (1) Although the human PrP transcript does not include the untranslated exon 2 found in its mouse and sheep counterparts, the large intron of the human PrP gene contains an exon 2-like sequence flanked by consensus splice acceptor and donor sites. (2) The mouse Prnpa but not the Prnpb allele found in 44 inbred lines contains a 6593 nucleotide retroviral genome inserted into the anticoding strand of intron 2. This intracisternal A-particle element is flanked by duplications of an AAGGCT nucleotide motif. (3) We found that the PrP gene regions contain from 40% to 57% genome-wide repetitive elements that independently increased the size of the locus in all three species by numerous mutations. The unusually long sheep PrP 3'-untranslated region contains a "fossil" 1.2-kb mariner transposable element. (4) We identified sequences in noncoding DNA that are conserved between the three species and may represent biologically functional sites.

Amino Acid Sequence↗

PubMed1998

Echolocation range of captive and free-ranging baiji (Lipotes vexillifer), finless porpoise (Neophocaena phocaenoides), and bottlenose dolphin (Tursiops truncatus).

The interclick intervals of captive dolphins are known to be longer than the two-way transit time between the dolphin and a target. In the present study, the interclick intervals of free-ranging baiji, finless porpoises, and bottlenose dolphins in the wild and in captivity were compared. The click intervals in open waters ranged up to 100-200 ms, whereas the click intervals in captivity were in the order of 4-28 ms. Echolocation of free-ranging dolphins appears to adapt to various distance in navigation or ranging, sometimes up to 140 m. Additionally, the difference of waveform characteristics of clicks between species was recognized in the frequency of maximum energy and the click duration.

Animals↗

PubMed1998

Adenovirus internalization and infection require dynamin.

The cell receptors that facilitate adenovirus internalization into cells have been identified; however, the infectious pathway of virus entry has not been established. Adenovirus entry and infection were examined in HeLa cells lacking or overexpressing mutant dynamin, a protein that specifically regulates clathrin-mediated endocytosis. Expression of mutant dynamin significantly reduced adenovirus internalization and gene delivery, indicating a functional requirement for this molecule. These findings are consistent with virus entry via the clathrin-coated pit pathway.

Adenoviruses, Human↗

PubMed1998

Mutagenesis of the BH3 domain of BAX identifies residues critical for dimerization and killing.

The BCL-2 family of proteins is comprised of proapoptotic as well as antiapoptotic members (S. N. Farrow and R. Brown, Curr. Opin. Genet. Dev. 6:45-49, 1996). A prominent death agonist, BAX, forms homodimers and heterodimerizes with multiple antiapoptotic members. Death agonists have an amphipathic alpha helix, called BH3; however, the initial assessment of BH3 in BAX has yielded conflicting results. Our BAX deletion constructs and minimal domain constructs indicated that the BH3 domain was required for BAX homodimerization and heterodimerization with BCL-2, BCL-XL, and MCL-1. An extensive site-directed mutagenesis of BH3 revealed that substitutions along the hydrophobic face of BH3, especially charged substitutions, had the greatest affects on dimerization patterns and death agonist activity. Particularly instructive was the BAX mutant mIII-1 (L63A, G67A, L70A, and M74A), which replaced the hydrophobic face of BH3 with alanines, preserving its amphipathic nature. BAXmIII-1 failed to form heterodimers or homodimers by yeast two-hybrid or immunoprecipitation analysis yet retained proapoptotic activity. This suggests that BAX's killing function reflects mechanisms beyond its binding to BCL-2 or BCL-XL to inhibit them or simply displace other protein partners. Notably, BAXmIII-1 was found predominantly in mitochondrial membranes, where it was homodimerized as assessed by homobifunctional cross-linkers. This characteristic of BAXmIII-1 correlates with its capacity to induce mitochondrial dysfunction, caspase activation, and apoptosis. These data are consistent with a model in which BAX death agonist activity may require an intramembranous conformation of this molecule that is not assessed accurately by classic binding assays.

Amino Acid Sequence↗

PubMed1998

A spectrum of mutations in the polycystic kidney disease-2 (PKD2) gene from eight Canadian kindreds.

Autosomal dominant polycystic kidney disease (ADPKD) is a common Mendelian disorder that affects approximately 1 in 1000 live births. Linkage studies have shown that the majority (approximately 85%) of cases are due to mutations in PKD1 on chromosome 16p, while mutations in PKD2 on chromosome 4q account for most of the remaining cases. Locus heterogeneity in ADPKD is known to contribute to differences in disease severity, with PKD1-linked families having earlier onset of end-stage renal disease (ESRD) than PKD2-linked families (mean age at ESRD: 56 versus 70, respectively). In this study, 11 Canadian families with ADPKD were screened for PKD2 mutations. In four families, linkage to PKD2 was previously documented. In the remaining seven smaller families, one or more affected members had late-onset ESRD at age 70 or older. Using single-stranded conformational polymorphism analysis, one affected member from each family was screened for mutations in all 15 exons of PKD2, which were PCR-amplified from genomic templates. A spectrum of mutations was found in approximately 73% (8 of 11) of the families screened, with no difference in the detection rate between the PKD2-linked families and the families with late-onset ESRD. In three unrelated families, insertion or deletion of an adenosine in a polyadenosine tract (i.e., (A)8 at nt 2152-2159) was found on exon 11, suggesting that this mononucleotide repeat tract is prone to mutations from "slipped strand mispairing." All mutations, scattered between exons 1 and 11, are predicted to result in a truncated polycystin 2 that lacks both the calcium-binding EF-hand domain and the two cytoplasmic domains required for the interaction of polycystin 2 with polycystin 1 and with itself. Furthermore, no correlation was found between the location of the mutations in the PKD2 coding sequence and disease severity. Thus, these findings are consistent with other recently published reports and suggest that most PKD2 mutations are inactivating.

Adolescent↗