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Biomedical subjects

B Wei

Publications and source records attributed to B Wei.

At least 19 recordsLinked to original sources

Supercooling of aqueous NaCl and KCl solutions under acoustic levitation.

The supercooling capability of aqueous NaCl and KCl solutions is investigated at containerless state by using acoustic levitation method. The supercooling of water is obviously enhanced by the alkali metal ions and increases linearly with the augmentation of concentrations. Furthermore, the supercooling depends on the nature of ions and is 2-3 K larger for NaCl solution than that for KCl solution in the present concentration range: Molecular dynamics simulations are performed to reveal the intrinsic correlation between supercoolability and microstructure. The translational and orientational order parameters are applied to quantitatively demonstrate the effect of ionic concentration on the hydrogen-bond network and ice melting point. The disrupted hydrogen-bond structure determines essentially the concentration dependence of supercooling. On the other hand, the introduced acoustic pressure suppresses the increase of supercooling by promoting the growth and coalescence of microbubbles, the effective nucleation catalysts, in water. However, the dissolved ions can weaken this effect, and moreover the degree varies with the ion type. This results in the different supercoolability for NaCl and KCl solutions under the acoustic levitation conditions.

Cold Temperature↗

TIMP-2 upregulates RECK expression via dephosphorylation of paxillin tyrosine residues 31 and 118.

We previously demonstrated that TIMP-2 increases the association of Crk with C3G and via subsequent activation of Rap1 enhances the expression of RECK, a membrane-anchored MMP inhibitor. In the present study, we investigate the mechanism of how the TIMP-2 signal is transduced from the alpha3beta1 integrin receptor to the Crk-C3G-Rap1 molecular complex. TIMP-2 treatment of human microvascular endothelial cells (hMVECs) increased the phosphorylation levels of Src at Tyr-527, the negative regulatory site, through enhanced association of Src with Csk. This results in the reduction of Src kinase activity and dephosphorylation of paxillin at Tyr-31/118, the target sites for Src kinase phosphorylation and also the binding sites for the downstream effector Crk. Such TIMP-2 effects accompany the disassembly of paxillin-Crk-DOCK180 molecular complex and, in turn, Rac1 inactivation. On the contrary, levels of paxillin-Crk-C3G complex formation are not reduced, rather slightly increased, which is consistent with our previous finding. Therefore, TIMP-2-mediated inhibition of Src kinase activity leads to the signaling switch from Rac1 to Rap1, thereby leading to enhanced RECK expression.

Cells, Cultured↗

Stable transformation of CHO Cells and human NARP cybrids confers oligomycin resistance (oli(r)) following transfer of a mitochondrial DNA-encoded oli(r) ATPase6 gene to the nuclear genome: a model system for mtDNA gene therapy.

Point and deletion mutations and a general depletion of mammalian mitochondrial DNA (mtDNA) give rise to a wide variety of medical syndromes that are refractory to treatment, possibly including aging itself. While gene therapy directed at correcting such deficits in the mitochondrial genome may offer some therapeutic benefits, there are inherent problems associated with a direct approach. These problems are primarily due to the high mitochondrial genome copy number in each cell and the mitochondrial genome being "protected" inside the double-membrane mitochondrial organelle. In an alternative approach there is evidence that genes normally present in the mitochondrial genome can be incorporated into the nuclear genome. To extend such studies, we modified the Chinese Hamster Ovary (CHO) mtDNA-located ATPase6 gene (possessing a mutation which confers oligomycin resistance- oli(r)) by altering the mtDNA code to the universal code (U-code) to permit the correct translation of its mRNA in the cytoplasm. The U-code construct was inserted into the nuclear genome (nucDNA) of a wild type CHO cell. The expressed transgene products enabled the transformed CHO cell lines to grow in up to 1000 ng mL(-1) oligomycin, while untransformed sensitive CHO cells were eliminated in 1 ng mL(-1) oligomycin. This approach, termed allotopic expression, provides a model that may make possible the transfer of all 13 mtDNA mammalian protein-encoding genes to the nucDNA, for treatments of mtDNA disorders. The CHO mtATPase6 protein is 85% identical to both the mouse and human mtATPase6 protein; these proteins are highly conserved in the region of the oligomycin resistance mutation. They are also well conserved in the regions of the oligomycin resistance mutation of the mouse, and in the region of a mutation found in Leigh's syndrome (T8993G), also called NARP (neurogenic weakness, ataxia, retinitis pigmentosum). It is likely that the CHO oli(r) mtATPase6 Ucode construct could impart oligomycin-resistance in human and mouse cells, as well as function in place of the mutant ATPase subunit in a NARP cell line. Preliminary experiments on human cybrids homoplasmic for the NARP mutation (kindly supplied by D.C. Wallace), transformed with our construct, display an increased oligomycin resistance that supports these suppositions.

Animals↗

Dynamics of acoustically levitated disk samples.

The acoustic levitation force on disk samples and the dynamics of large water drops in a planar standing wave are studied by solving the acoustic scattering problem through incorporating the boundary element method. The dependence of levitation force amplitude on the equivalent radius R of disks deviates seriously from the R3 law predicted by King's theory, and a larger force can be obtained for thin disks. When the disk aspect ratio gamma is larger than a critical value gamma(*) ( approximately 1.9 ) and the disk radius a is smaller than the critical value a(*) (gamma) , the levitation force per unit volume of the sample will increase with the enlargement of the disk. The acoustic levitation force on thin-disk samples ( gamma</= gamma(*) ) can be formulated by the shape factor f(gamma,a) when a</= a(*) (gamma) . It is found experimentally that a necessary condition of the acoustic field for stable levitation of a large water drop is to adjust the reflector-emitter interval H slightly above the resonant interval H(n) . The simulation shows that the drop is flattened and the central parts of its top and bottom surface become concave with the increase of sound pressure level, which agrees with the experimental observation. The main frequencies of the shape oscillation under different sound pressures are slightly larger than the Rayleigh frequency because of the large shape deformation. The simulated translational frequencies of the vertical vibration under normal gravity condition agree with the theoretical analysis.

Journal Article↗

Removal of fluoride from water by carbon nanotube supported alumina.

This paper describes a new adsorption material carbon nanotube supported alumina, used for fluoride removal from water. The effects of calcination temperature, alumina loading and pH on fluoride removal capacity were investigated. The optimum calcination temperature for preparing the adsorbents is 450 degrees C and the largest adsorption capacity takes place at alumina loading of 30 wt%. The broad pH range of 6.0-9.0 with high adsorption capability suggests that carbon nanotube supported alumina have great potential applications in practice. The adsorption data at different pH were well fitted to the Freundlich isotherm. The kinetic studies show that the adsorption reaction of fluoride removal by carbon nanotube supported alumina can be described by a second orderrate equation.

Adsorption↗

Eutectic growth under acoustic levitation conditions.

Samples of Pb-Sn eutectic alloy with a high density of 8.5 x 10(3) kg/m(3) are levitated with a single-axis acoustic levitator, and containerlessly melted and then solidified in argon atmosphere. High undercoolings up to 38 K are obtained, which results in a microstructural transition of "lamellas-broken lamellas-dendrites." This transition is further investigated in the light of the coupled zone for eutectic growth and the effects of ultrasound. The breaking of regular eutectic lamellas and suppression of gravity-induced macrosegregation of (Pb) and (Sn) dendrites are explained by the complicated internal flow inside the levitated drop, which is jointly induced by the shape oscillation, bulk vibration and rotation of the levitated drop. The ultrasonic field is also found to drive forced surface vibration, which subsequently excites capillary ripples and catalyzes nucleation on the sample surface.

Journal Article↗

Dependence of acoustic levitation capabilities on geometric parameters.

A two-cylinder model incorporating boundary element method simulations is developed, which builds up the relationship between the levitation capabilities and the geometric parameters of a single-axis acoustic levitator with reference to wavelength. This model proves to be successful in predicting resonant modes of the acoustic field and explaining axial symmetry deviation of the levitated samples near the reflector and emitter. Concave reflecting surfaces of a spherical cap, a paraboloid, and a hyperboloid of revolution are investigated systematically with regard to the dependence of the levitation force on the section radius R(b) and curvature radius R (or depth D) of the reflector. It is found that the levitation force can be remarkably enhanced by choosing an optimum value of R or D, and the possible degree of this enhancement for spherically curved reflectors is the largest. The degree of levitation force enhancement by this means can also be facilitated by enlarging R(b) and employing a lower resonant mode. The deviation of the sample near the reflector is found likely to occur in case of smaller R(b), larger D, and a higher resonant mode. The calculated dependence of levitation force on R, R(b), and the resonant mode is also verified by experiment and finally demonstrated to be in good agreement with experimental results, in which considerably a strong levitation force is achieved to levitate an iridium sphere which has the largest density of 22.6 g/cm(3).

Journal Article↗

Levitation of iridium and liquid mercury by ultrasound.

Single-axis acoustic levitation of the heaviest solid (iridium, rho=22.6 g cm(-3)) and liquid (mercury, rho=13.6 g cm(-3) on the Earth is achieved by greatly enhancing both the levitation force and stability through optimizing the geometric parameters of the levitator. The acoustically levitated Pb-Sn eutectic alloy melt (rho=8.5 g cm(-3)) is highly undercooled by up to 38 K, which results in a microstructural transition of "lamellae-broken lamellae-dendrites." The drastic enhancement of levitation capability indicates a broader application range of single-axis acoustic levitation.

Journal Article↗

Activation of the Jnk signaling pathway by a dual-specificity phosphatase, JSP-1.

The mitogen-activated protein kinases (MAPKs) are integral to the mechanisms by which cells respond to physiological stimuli, such as growth factors, hormones, and cytokines, and to a wide variety of environmental stresses. The MAPKs, which are stimulated by phosphorylation of a TXY motif in their activation loop, are components of signal transduction cascades in which sequential activation of protein kinases culminates in their activation and their subsequent phosphorylation of various effector proteins that mediate the physiological response. MAPKs are also subject to dephosphorylation and inactivation, both by enzymes that recognize the residues of the TXY motif independently and by dual specificity phosphatases, which dephosphroylate both Tyr and Ser/Thr residues. We report the identification and characterization of a novel dual specificity phosphatase. Contrary to expectation, this broadly expressed enzyme did not inactivate MAPKs in transient cotransfection assays but instead displayed the capacity to function as a selective activator of the MAPK Jnk, hence the name, Jnk Stimulatory Phosphatase-1 (JSP-1). This study illustrates a new aspect of the regulation of MAPK-dependent signal transduction and raises the possibility that JSP-1 may offer a different perspective to the study of various inflammatory and proliferative disorders associated with dysfunctional Jnk signaling.

Amino Acid Sequence↗

Subunit communication in tetrameric class 2 human liver aldehyde dehydrogenase as the basis for half-of-the-site reactivity and the dominance of the oriental subunit in a heterotetramer.

Data has been published showing that in heterotetrameric liver mitochondrial aldehyde dehydrogenase composed of the active (E487) and the inactive Oriental-variant (K487) subunit, the Oriental variant was dominant and caused the inactivation of the E487 subunit. The published structures of the enzyme showed that the glutamate at position 487 is salt bonded to an arginine (475) in a different subunit. Arg475 was mutated to a glutamine to test for its importance in causing the Oriental variant to be an enzyme with a high Km for NAD and a low specific activity. Unexpectedly, the R475Q mutant exhibited positive cooperativity in NAD binding with a Hill coefficient of 2. Individual heterotetramers composed of subunits of E487 and K487 were produced by making changes to two residues on the surface of the enzyme and then co-expressing both cDNAs in E. coli. The E(3)K form had essentially 50% the activity of the E(4) homotetrameric form while EK(3) had essentially the same properties as did the homotetrameric K(4) Oriental variant. This showed that in a dimer pair composed of one K- and one E- subunit the K-subunit became dominant and caused the inactivation of its E-partner. Further, pre-steady state burst data and steady state kinetic data make it appear that there was one functioning active subunit in each of the dimer pairs that made up the tetrameric enzyme. Thus, the half-of-the-site reactivity is a result of having one functioning and one non-functioning subunit in each dimer pair. The actual structural basis for this is still not understood, but could be related to the E487-R475 inter-dimer salt bond.

Aldehyde Dehydrogenase↗

Making an Oriental equivalent of the yeast cytosolic aldehyde dehydrogenase as well as making one with positive cooperativity in coenzyme binding by mutations of glutamate 492 and arginine 480.

Yeast has at least three partially characterized aldehyde dehydrogenases. Previous studies by gene disrupted in our laboratory revealed that the Saccharomyces cerevisiae cytosol ALDH1 played an important role in ethanol metabolism as did the class 2 mitochondrial enzyme. To date, few mutagenesis studies have been performed with the yeast enzymes. An important human variant of ALDH is one found in Asian People. In it, the glutamate at position 487 is replaced by a lysine. This glutamate interacts with an arginine (475) that is located in the subunit that makes up the dimer pair in the tetrameric enzyme. Sequence alignment shows that these two residues are located at positions 492 and 480, respectively, in the yeast class 1 enzyme which shares just 45% sequence identity with the human enzymes. Mutating glutamate 492 to lysine produced an enzyme with altered kinetic properties when compared to the wild-type glutamate-enzyme. The K(m) for NADP of E492K increased to nearly 3600 microM compare to 40 microM for wild-type enzyme. The specific activity decreased more than 10-fold with respect to the recombinant wild-type yeast enzyme. Moreover, substituting a glutamine for a glutamate was not detrimental in that the E492Q had wild-type-like K(m) for NADP and V(max). These properties were similar to the changes found with the human class 2 E487K mutant form. Further, mutating arginine 480 to glutamine produced an enzyme that exhibited positive cooperativity in NADP binding. The K(m) for NADP increased 11-fold with a Hill coefficient of 1.6. The NADP-dependent activity of R480Q mutant was 60% of wild-type enzyme. Again, these results are very similar to what we recently showed to occur with the human enzyme [Biochemistry 39 (2000) 5295-5302]. These findings show that the even though the glutamate and arginine residues are not conserved, similar changes occur in both the human and the yeast enzyme when either is mutated.

Aldehyde Dehydrogenase↗

Using rigid motion constraints for the registration of free-form surfaces.

We present a new method for registration of freeform surfaces based on the iterative closest point (ICP) method and on geometric properties of reflected correspondence vectors. The method is based on computing relative gaps between reflected correspondences and between the projections of reflected correspondences along the rotation axis and using these to eliminate false matches. Experimental results on synthetic data and on real range images demonstrate that the method is robust and accurate for image registration with small motions.

Algorithms↗

Use of artificial neural network in the prediction of algal blooms.

A model to quantify the interactions between abiotic factors and algal genera in Lake Kasumigaura, Japan was developed using artificial neural network technology. Results showed that the timing and magnitude of algal blooms of Microcystis, Phormidium and Synedra in Lake Kasumigaura could be successfully predicted. As for the newly occurring dominant Oscillatoria, results were not satisfactory. The evaluation of the importance of factors showed that Microcystis, Phormidium, Oscillatoria and Synedra were alkalophilic. The algal proliferation for Microcystis, Oscillatoria and Synedra decrease due to the increase in total nitrogen, while the growth of Phormidium is enhanced with more nitrogen. In addition, the algal density is affected by zooplankton grazing but with the exception of Phormidium due to it being poor food source. Algal responses to the orthogonal combinations of the external environmental factors, chemical oxygen demand, pH, total nitrogen and total phosphorus at three levels were modeled. Various combinations of environmental factors enhance the proliferation of some algae while other combinations inhibit bloom formation.

Cyanobacteria↗

Effects of environmental factors on the seasonally change of phytoplankton populations in a closed freshwater pond.

The effect of environmental factors affecting the population dynamic of phytoplankton population was studied seasonally in a closed freshwater pond with the natural water of Lake Kasumigaura, Japan. The predominant phytoplankton species in the pond were classified as Oscillatoria tenuis, Synedra ulna, Chlamydomonas cingulata and Cyclotella kutzingiana, with dominant species of Melosira italica, Synechococcus sp. and Cryptomonas ovata. The most important limiting factor for filament length and cell size of most of the phytoplankton community was temperature. Availability of most nutrients had profound effects on the phytoplankton growth as nonlimiting factors. Based on our results, it is suggested that cell cycle progression was also temporally regulated by a multiplicity of external factors such as temperature, nutrients, dissolved oxygen (DO) and pH. At high pH and high temperature, consistent increase of cell division of some phytoplanketers such as Synedra, Cyclotella, Chlamydomonas and Melosira occurs, and there also may be species-specific responses for other species of phytoplankters. In addition, the cell divisions of phytoplankters are differentially regulated in different phytoplankton species.

Cell Division↗

The Crohn's disease-associated bacterial protein I2 is a novel enteric t cell superantigen.

An aberrant T cell response to enteric bacteria is important in inflammatory bowel disease. However, the identity of relevant microbial antigens is unknown. Here, we report the presence of I2, a Crohn's disease-associated microbial gene, in the murine intestine. The I2 protein induced a proliferative and IL-10 response by CD4(+) T cells from unimmunized mice. The I2 response was dependent on MHC class II-mediated recognition but did not require antigen processing. Selective activation was observed for the TCR-Vbeta5 subpopulation. These findings indicate that the I2 protein is a new class of T cell superantigen and suggest that colonization by the I2 microorganism in susceptible hosts may provide a superantigenic stimulus pertinent to Crohn's disease pathogenesis.

Amino Acid Sequence↗

Molecular cloning of a Bacteroides caccae TonB-linked outer membrane protein identified by an inflammatory bowel disease marker antibody.

Commensal enteric bacteria are a required pathogenic factor in inflammatory bowel disease (IBD), but the identity of the pertinent bacterial species is unresolved. Using an IBD-associated pANCA monoclonal antibody, a 100-kDa protein was recently characterized from an IBD clinical isolate of Bacteroides caccae (p2Lc3). In this study, consensus oligonucleotides were designed from 100-kDa peptides and used to identify a single-copy gene from the p2Lc3 genome. Sequence analysis of the genomic clone revealed a 2,844-bp (948 amino acid) open reading frame encoding features typical of the TonB-linked outer membrane protein family. This gene, termed ompW, was detected by Southern analysis only in B. caccae and was absent in other species of Bacteroides and gram-negative coliforms. The closest homologues of OmpW included the outer membrane proteins SusC of Bacteroides thetaiotaomicron and RagA of Porphyromonas gingivalis. Recombinant OmpW protein was immunoreactive with the monoclonal antibody, and serum anti-OmpW immunoglobulin A levels were elevated in a Crohn's disease patient subset. These findings suggest that OmpW may be a target of the IBD-associated immune response and reveal its structural relationship to a bacterial virulence factor of P. gingivalis and periodontal disease.

Amino Acid Sequence↗

Vascular injury induces posttranscriptional regulation of the Id3 gene: cloning of a novel Id3 isoform expressed during vascular lesion formation in rat and human atherosclerosis.

The molecular mechanisms that regulate the proliferation of smooth muscle cells (SMCs) of the vasculature in response to injury are poorly understood. Members of the inhibitor of DNA binding (Id) class of helix-loop-helix transcription factors are known to regulate the growth of a variety of cell types; however, the expression of the various Id genes in SMCs and in vascular lesions has not been examined. In the present study, the yeast 2-hybrid system was used to clone Id genes from a cultured rat aortic SMC library. By use of ubiquitous E proteins as bait, Id3 and a novel isoform of Id3 (Id3a) were cloned. Id3a is the product of alternative splicing of the Id3 gene, resulting in inclusion of a 115-bp "coding intron," which encodes a unique 29-amino acid carboxyl terminus for the Id3a protein. Unlike Id3, Id3a mRNA was not detected in the normal rat carotid artery. However, after balloon injury, Id3a was abundantly expressed throughout the neointimal layer. In addition, mRNA of the human homologue of Id3a (Id3L) was detected in human carotid atherosclerotic plaques. Adenovirus-mediated overexpression of these Id3 isoforms in cultured rat aortic SMCs revealed that infection of SMCs with an adenovirus overexpressing Id3a (in contrast to Id3) resulted in a significant decrease in cell number versus AdLacZ-infected cells. DNA fragmentation analysis suggested that this decrease in SMC viability was due to increased apoptotic activity in cells infected with adenovirus overexpressing Id3a. These results provide evidence that alternative splicing of the Id3 gene may represent an important mechanism by which neointimal SMC growth is attenuated during vascular lesion formation.

Adenoviridae↗

[Clinicopathologic analysis of ovarian serous and mucinous borderline tumors].

OBJECTIVE: To understand the clinicopathological features of ovarian borderline tumors and evaluate the significance of various histological appearances. METHODS: Forty-five patients with borderline tumors were analyzed retrospectively. The stage distribution was stage I, 34 patients, stage II, 4 patients and stage III, 7 patients. RESULTS: Forty-five cases of borderline tumors accounted for 25.4% of ovarian epithelial malignant tumors. The ratio of serous borderline tumors (SBTs) to mucinous borderline tumors (MBTs) was approximately 1:1.3. Nine of eleven SBTs with exophytic growth presented peritoneal implants, of which two were invasive and seven noninvasive. Two SBTs and 1 MBT recurred after 5, 4, and 1 years respectively. Thirty-three cases were followed from 2 to 9 years. The 5-year survival rate was 100% (Kaplan-Meir method). CONCLUSIONS: The prognosis of the SBTs is favorable. SBTs with exophytic growth are often associated with peritoneum implants.

Adenocarcinoma, Mucinous↗