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Biomedical subjects

P Wu

Publications and source records attributed to P Wu.

At least 19 recordsLinked to original sources

Fluorescence-based monitoring of tracer and substrate distribution in an UASB reactor.

In this work, rhodamine-related fluorescence was measured on-line at four reactor heights in order to study hydrodynamics within an upflow anaerobic sludge bed reactor. A linear dependence of the dispersion coefficient (D) on the upflow velocity was observed, while the influence of the organic loading rate (OLR) was insignificant. Furthermore, the Bodenstein number of the reactor loaded with granulated sludge was found to be position-dependent with the largest values measured at the bottom of the sludge bed. This trend was not observed in the reactor without sludge. Chemical oxygen demand (COD) and volatile fatty acid (VFA) concentrations were measured at the same reactor heights as in rhodamine tests using conventional off-line analytical methods and on-line multiwavelength fluorometry. Significant spatial COD and VFA gradients were observed at organic loading rates above 6g COD l(R)(-1)d(-1) and linear upflow velocities below 0.8m h(-1).

Bioreactors↗

Gene expression profiling and analysis of signaling pathways involved in priming and differentiation of human neural stem cells.

Human neural stem cells have the ability to differentiate into all three major cell types in the CNS including neurons, astrocytes and oligodendrocytes. The multipotency of human neural stem cells shed a light on the possibility of using stem cells as a therapeutic tool for various neurological disorders including neurodegenerative diseases and neurotrauma that involve a loss of functional neurons. We have discovered previously a priming procedure to direct primarily cultured human neural stem cells to differentiate into almost pure neurons when grafted into adult CNS. However, the molecular mechanism underlying this phenomenon is still unknown. To unravel transcriptional changes of human neural stem cells upon priming, cDNA microarray was used to study temporal changes in human neural stem cell gene expression profile during priming and differentiation. As a result, transcriptional levels of 520 annotated genes were detected changed in at least at two time points during the priming process. In addition, transcription levels of more than 3000 hypothetical protein encoding genes and EST genes were modulated during the priming and differentiation processes of human neural stem cells. We further analyzed the named genes and grouped them into 14 functional categories. Of particular interest, key cell signal transduction pathways, including the G-protein-mediated signaling pathways (heterotrimeric and small monomeric GTPase pathways), the Wnt signaling pathway and the TGF-beta pathway, are modulated by the neural stem cell priming, suggesting important roles of these key signaling pathways in priming and differentiation of human neural stem cells.

Bone Morphogenetic Proteins↗

Inhibition of telomerase enhances apoptosis induced by sodium butyrate via mitochondrial pathway.

Telomerase activation represents an early step in carcinogenesis. Increased telomerase activity in cervical cancer suggests a potential target for the development of novel therapeutic drugs. The aim of this study is to investigate the impact of telomerase activity on the biological features of HeLa cells and the possible mechanisms of enhanced apoptosis rate induced by sodium butyrate after telomerase inhibition. We introduced vectors encoding dominate negative (DN)-hTERT, wild-type (WT)-hTERT, or a control vector expressing only a drug-resistance marker into HeLa cells. Thus we assessed the biological effects of telomerase activity on telomere length, cell proliferation, chemosensitivity and radiosensitivity. In order to understand the mechanisms in which DN-hTERT enhances the apoptosis induced by sodium butyrate, we detected the release status of cytochrome c and apoptosis inducing factor (AIF) from mitochondria. Ectopic expression of DN-hTERT resulted in inhibition of telomerase activity, reduction of telomere length, decreased colony formation ability, and loss of tumorigenicity in nude mice. Moreover, DN-hTERT transfected HeLa cells with shortened telomeres were more susceptible to multiple chemotherapeutic agents and radiation. WT-hTERT transfected HeLa cells with longer telomeres exhibited resistance to radiation and chemotherapeutic agents. Our data demonstrate that elevated release level of cytochrome c and AIF from mitochondria might contribute to the enhanced apoptosis in DN-hTERT transfected HeLa cells after treatment with sodium butyrate. Inhibition of telomerase might serve as a promising adjunctive therapy combined with conventional therapy in cervical cancer.

Animals↗

Renal injury: similarities and differences in male and female rats with the metabolic syndrome.

The metabolic syndrome is complicated by nephropathy in humans and rats, and males are more affected than females. We hypothesized that female rats had reduced expression of glomerular oxidized low-density lipoprotein (oxLDL) receptor 1 (LOX-1), attendant glomerular oxidant injury, and renal inflammation. Three groups, obese males (OM), obese females (OF), and lean males (LM) of first-generation (F(1)) hybrid rats derived from the Zucker fatty diabetic (ZDF) strain and the spontaneous hypertensive heart failure rat (SHHF/Gmi-fa) were studied from 6 to 41 weeks of age. OM had severe renal oxidant injury and renal failure. Their glomeruli expressed the LOX-1, and exhibited heavier accumulation of the lipid peroxide 4-hydroxynonenal (4-HNE). OM had compromised mitochondrial enzyme function, more renal fibrosis, and vascular leakage. Younger LM, OM, and OF ZS (ZDF/SHHF F(1) hybrid rat) rats, studied from 6 to 16 weeks of age, showed that unutilized renal lipids were comparable in OM and OF, although young OM had worse nephropathy and inflammation. In conclusion, glomerular LOX-1 expression is coupled to deposits of 4-HNE and glomerulosclerosis in OM. We presume that LOX-1 enhances glomerular uptake of oxidized lipids and renal inflammation, causing greater oxidant stress and severe glomerulosclerosis. In OF, renal protection from lipid oxidants appears to be conferred by blunted glomerular LOX-1 expression and renal inflammation.

Animals↗

Human neural stem cell-derived cholinergic neurons innervate muscle in motoneuron deficient adult rats.

Motoneuron damage occurs in spinal cord injury and amyotrophic lateral sclerosis. Current advances offer hope that human embryonic stem cells [Science 282 (1998) 1145] or neural stem cells (NSC) [Exp Neurol 161 (2000) 67; Exp Neurol 158 (1999) 265; J Neurosci Methods 85 (1998) 141; Proc Natl Acad Sci USA 97 (2000) 14720; Exp Neurol 156 (1999) 156 ] may be donors to replace lost motoneurons. Previously, we developed a priming procedure that produced cholinergic cells that resemble motoneurons from human NSCs grafted into adult rat spinal cord [Nat Neurosci 5 (2002a) 1271]. However, effective replacement therapy will ultimately rely on successful connection of new motoneurons with their muscle targets. In this study, we examined the potential of human fetal NSC transplantation to replace lost motoneurons in an animal model of chronic motoneuron deficiency (newborn sciatic axotomy) [J Comp Neurol 224 (1984) 252; J Neurobiol 23 (1992) 1231]. We found, for the first time, that human neural stem cell-derived motoneurons send axons that pass through ventral root and sciatic nerve to form neuromuscular junctions with their peripheral muscle targets. Furthermore, this new cholinergic innervation correlates with partial improvement of motor function.

Animals↗

A new antihistamine levocetirizine inhibits eosinophil adhesion to vascular cell adhesion molecule-1 under flow conditions.

BACKGROUND: We previously demonstrated that low concentrations of a new antihistamine levocetirizine inhibited eosinophil transmigration through human microvascular endothelial cells. OBJECTIVE: Here, the inhibitory effect of levocetirizine on eosinophil adhesion to recombinant human vascular cell adhesion molecule-1 (rhVCAM)-1 was examined under conditions of shear stress using an in vitro model of the post-capillary venules. METHODS: Eosinophils isolated from normal subjects were pre-incubated with a concentration range of levocetirizine (10(-6)-10(-10) m) or negative dilution control. Resting or granulocyte macrophage-colony stimulating factor (GM-CSF)-stimulated cells were pumped through rhVCAM-1 (10 microg/mL) coated capillary tubes using a microfluidic syringe pump at a precise and constant flow rate (1 dyn/cm(2)). Images of rolling and firmly adherent eosinophils were captured using real-time video microscopy. RESULTS: Levocetirizine significantly inhibited resting eosinophil adhesion to rhVCAM-1 with maximal effect at 10(-8) M with an EC(50) of 10(-9) m. Levocetirizine almost abolished resting eosinophil adhesion by the 15 min time-point. GM-CSF significantly enhanced eosinophil adhesion and their ability to flatten on rhVCAM-1. Both phenomena were inhibited by levocetirizine in a dose-dependent manner, at both 5 and 15 min (optimal concentration of 10(-8) m with an EC(50) of 10(-9) m). Real-time imaging revealed that the effect of levocetirizine on post-adhesion behaviour (detachment, flatness) contributed to its inhibitory action on eosinophil adhesion to rhVCAM-1. In contrast, very late antigen (VLA)-4 mAb inhibited eosinophil adhesion to rhVCAM-1 from the earliest time-points. CONCLUSION: Physiologically relevant concentrations of levocetirizine inhibit resting and GM-CSF-stimulated firm eosinophil adhesion to rhVCAM-1 under flow conditions.

Antibodies, Monoclonal↗

K2[HCr2AsO10]: redetermination of phase II and the predicted structure of phase I.

Our prediction that phase II of dipotassium hydrogen chromatoarsenate, K(2)[HCr(2)AsO(10)], is ferroelectric, based on the analysis of the atomic coordinates by Averbuch-Pouchot, Durif & Guitel [Acta Cryst. (1978), B34, 3725-3727], led to an independent redetermination of the structure using two separate crystals. The resulting improved accuracy allows the inference that the H atom is located in the hydrogen bonds of length 2.555 (5) angstroms which form between the terminal O atoms of shared AsO(3)OH tetrahedra in adjacent HCr(2)AsO(10)(2-) ions. The largest atomic displacement of 0.586 angstroms between phase II and the predicted paraelectric phase I is by these two O atoms. The H atoms form helices of radius approximately 0.60 A about the 3(1) or 3(2) axes. Normal probability analysis reveals systematic error in seven or more of the earlier atomic coordinates.

Journal Article↗

Phase transitions in K2Cr2O7 and structural redeterminations of phase II.

Crystals of phase II K2Cr2O7, potassium dichromate, space group P1 , grown from aqueous solution undergo a first-order transition to phase I, space group reportedly P21/n, at a phase-transition temperature, TPT, of 544 (2) K on first heating; the corresponding transition on cooling is at 502 (2) K. The endotherm on subsequent heatings occurs reproducibly at TPT = 531 (2) K. Mass loss between ca 531 and 544 K, identified as included water, is rapid and continues more slowly to higher temperatures for a total loss of ca 0.20%. The higher TPT on first heating is associated with the increasing pressure of superheated water occupying inclusion defects. The latent diagonal glide plane in phase II allows the structure of phase I to be inferred. The triclinic structure at 296 K has been independently redetermined. Normal probability analysis shows high consistency between the resulting and previous atomic coordinates, but with uncertainties reduced by a factor of ca 2. The earlier uncertainties are systematically underestimated by a comparable factor. The structure of phase IIb, space group A2/a on transposing axes, was determined at ca 300 K by Krivovichev et al. [Acta Cryst. (2000), C56, 629-630]. The first-order transition between phases I and II arises from the ca 60 degrees relative rotation of terminal O atoms in each tetrahedron as the n glide plane is gained or lost. A transition between phases IIb and I, also of first order, is likely but not between phases II and IIb. An intermediate phase may exist between phases IIb and I.

Journal Article↗

Analysis of transcripts that are differentially expressed in three sectors of the rice root system under water deficit.

Short periods of water deprivation can stimulate the growth of seminal and lateral roots in rice, and inhibit the emergence of adventitious roots. Identification of genes in the different tissues that respond to a water deficit may help us to understand the mechanism underlying root growth under conditions when water is scarce. cDNA-amplified fragment length polymorphism (AFLP) analysis was used to profile gene expression upon imposition of water deficit in three types of root tissue from the upland rice variety Azucena: seminal root tips, lateral root zones and adventitious root primordial zones. In all, 121 unique transcript-derived fragments (TDFs) were cloned, and Northern analysis was carried out for 30 TDFs to confirm their expression patterns. Sixty-six TDFs were differentially expressed in all three root samples. Four (AC2, D6, L22 and T23) were up-regulated by water deficit in seminal root tips and lateral root zones, and down-regulated in adventitious root primordial zones, an expression pattern which reflects the phenotypic changes observed in the different root sectors. In contrast, T17 and T37 showed the opposite expression pattern in Azucena: up-regulation in adventitious roots and repression in the other two zones. Functions could be assigned to five of these six TDFs on the basis of homology: they encode an expansin (T37), a fruit-ripening protein similar to ASR (T23), submergence-induced protein 2A (T17), a dehydrin (D6) and a 9- cis -epoxycarotenoid dioxygenase1 (L22), respectively. AC2 did not show a significant match to any known gene. Northern analysis showed that these six clones exhibited expression patterns that differed between the two cultivars tested (Azucena and the lowland variety IR1552) with respect to regulation by water limitation. Furthermore, T17, T37, D6 and T23 mapped within intervals known to contain QTLs (quantitative trait loci) for root growth in rice under water deficit. These genes may regulate or co-regulate the growth and development of the three root zones in a tissue-specific manner, and may play a role in the processes that underlie the early changes in root architecture under conditions of water deprivation.

Blotting, Northern↗

Prediction of apatite lattice constants from their constituent elemental radii and artificial intelligence methods.

Lattice constants (LCs) of all possible 96 apatite compounds, A(5)(BO(4))(3)C, constituted by A[double bond]Ba(2+), Ca(2+), Cd(2+), Pb(2+), Sr(2+), Mn(2+); B[double bond]As(5+), Cr(5+), P(5+), V(5+); and C[double bond]F(1-), Cl(1-), Br(1-), OH(1-), are predicted from their elemental ionic radii, using pattern recognition (PR) and artificial neural networks (ANN) techniques. In particular, by a PR study it is demonstrated that ionic radii predominantly govern the LCs of apatites. Furthermore, by using ANN techniques, prediction models of LCs a and c are developed, which reproduce well the measured LCs (R(2)=0.98). All the literature reported on 30 pure and 22 mixed apatite compounds are collected and used in the present work. LCs of all possible 66 new apatites (assuming they exist) are estimated by the developed ANN models. These proposed new apatites may be of interest to biomedical research especially in the design of new apatite biomaterials for bone remodeling. Similarly these techniques may also be applied in the study of interface growth behaviors involving other biomaterials.

Algorithms↗

Biological laser printing: a novel technique for creating heterogeneous 3-dimensional cell patterns.

We have developed a laser-based printing technique, called biological laser printing (BioLP). BioLP is a non-contact, orifice-free technique that rapidly deposits fL to nL scale volumes of biological material with spatial accuracy better than 5 microm. The printer's orifice-free nature allows for transfer of a wide range of biological material onto a variety of substrates. Control of transfer is performed via a computer-aided design/computer-aided manufacturing (CAD/CAM) system which allows for deposition rates up to 100 pixels of biological material per second using the current laser systems. In this article, we present a description of the apparatus, a model of the transfer process, and a comparison to other biological printing techniques. Further, examples of current system capabilities, such as adjacent deposition of multiple cell types, large-scale cell arrays, and preliminary experiments on creating multi-layer cell constructs are presented. These cell printing experiments not only demonstrate near 100% viability, they also are the first steps toward using BioLP to create heterogeneous 3-dimensional constructs for use in tissue engineering applications.

Animals↗

Toward immobilized antibody microarray optimization: print buffer and storage condition comparisons on performance.

Performance issues in protein arrays differ substantially from DNA microarrays because of the different intrinsic construction and stability of proteins versus DNA, and the impact on biological recognition phenomena required for assay from a surface-immobilized state. Maintenance of the capture bioactivity and performance of antibodies on microarray substrates for relatively long periods (under storage or shipping) requires carefully designed and optimized print buffer and storage conditions. We have focused on selection of several additives added to antibody print buffers with different concentrations, and storage of printed antibody slides under different temperatures from 1 day to 4 weeks. Activities of these slides were compared in capture assay formats in order to compare the effects of print buffer and storage conditions of antibody microarrays. Commercial polymer-coated Optarray slides exhibited better performance over Codelink slides in terms of activity of anti-human TNF alpha and IL-1 beta. IL-1 beta antibodies immobilized on both commercial surfaces showed much higher activity than TNF alpha antibodies, regardless of source or vendor. Addition of 1% (v/v) PEO to the print buffer increased the printed analyte capture activity of IL-1 beta on Optarray slides compared to standard print buffer without any additives. This helps to maintain antibody immobilized analyte capture activity for 4 weeks when stored at -70 degrees C.

Adsorption↗

Use of asymmetric somatic hybridization for transfer of the bacterial blight resistance trait from Oryza meyeriana L. to O. sativa L. ssp. japonica.

Bacterial blight is one of the major diseases affecting rice productivity. To improve the resistance of cultivated rice to bacterial blight, we introduced a bacterial blight resistance trait from Oryza meyeriana, a wild rice species, into an elite japonica rice cultivar (Dalixiang) using asymmetric somatic hybridization. One hundred and thirty-two independent lines were regenerated. The hybrid plants possessed several morphological features of the donor species, O. meyeriana. Random amplified polymorphic DNA analysis revealed that hybrid plants exhibited banding patterns derived from their parental genotypes. For the majority of the hybrids, resistance to bacterial blight pathogens was intermediate to that observed for O. meyeriana and O. sativa (cv. Dalixiang). Four of the hybrid lines exhibited a high bacterial blight resistance, but it was less than that observed for O. meyeriana. These results demonstrate that O. meyeriana can be used as a good genetic source for improving bacterial blight resistance in commercial rice cultivars through asymmetric somatic hybridization.

Crosses, Genetic↗

Mapping QTLs and candidate genes for rice root traits under different water-supply conditions and comparative analysis across three populations.

To investigate the genetic factors underlying constitutive and adaptive morphological traits of roots under different water-supply conditions, a recombinant inbred line (RIL) population derived from a cross between the lowland rice variety IR1552 and the upland rice variety Azucena with 249 molecular markers, was used in cylindrical-pot experiments. Eighteen QTLs were detected for seminal root length (SRL), adventitious root number (ARN), and lateral root length (LRL) and lateral root number (LRN) on the seminal root at a soil depth of from 3 to 6 cm under flooding and upland conditions. One identical QTL was detected under both flooding and upland conditions. The relative parameters under the two water-supply conditions were also used for QTL analysis. Five QTLs for upland induced variations in the traits were detected with the positive alleles from Azucena. A comparative analysis was performed for the QTLs detected in this study and those reported from two other populations with Azucena as a parent. Several identical QTLs for root elongation were found across the three populations with positive alleles from Azucena. Candidate genes were screened from ESTs and cDNA-AFLP clones for comparative mapping with the detected QTLs. Two genes for cell expansion, OsEXP2 and endo-1,4-beta-D-glucanase EGase, and four cDNA-AFLP clones from root tissues of Azucena, were mapped on the intervals carrying the QTLs for SRL and LRL under upland conditions, respectively.

Blotting, Northern↗

Adeno-associated viral transfer of opioid receptor gene to primary sensory neurons: a strategy to increase opioid antinociception.

To develop a genetic approach for the treatment of pain, we introduced a recombinant adeno-associated viral (rAAV) vector containing the cDNA for the mu-opioid receptor (muOR) into primary afferent neurons in dorsal root ganglia (DRGs) of rats, which resulted in a long-lasting (>6 months) increase in muOR expression in DRG neurons. The increase greatly potentiated the antinociceptive effects of morphine in rAAV-muOR-infected rats with and without inflammation. Perforated patch recordings indicated that the efficacy and potency of opioid inhibition of voltage-dependent Ca(2+) channels were enhanced in infected neurons, which may underlie the increase in opiate efficacy. These data suggest that transfer of opioid receptor genes into DRG cells with rAAV vectors may offer a new therapeutic strategy for pain management.

Animals↗

The causes of and cures for contact lens-induced peripheral ulcer.

PURPOSE: Contact lens-induced peripheral ulceration (CLPU) is a relatively common adverse response associated with wearing hydrogel lenses, especially on an extended wear schedule. Bacteriologic examination of lenses at the time of an event has demonstrated an association with Staphylococci spp. We sought to investigate the causes of CLPU in a rabbit model of contact lens wear. METHODS: Rabbits wore contact lenses for a period of 24 hr in the presence or absence of bacteria or in presence or absence of epithelial scratches made in the periphery of the cornea before lens wear. Bacteria tested were a strain of Staphylococcus aureus or a strain of Staphylococcus epidermidis isolated from human CLPUs. Rabbits were also challenged with S. aureus in the presence of an epithelial defect and in the absence of a contact lens. Corneas were monitored by slitlamp, histology, and microbial culture after 24 hr. RESULTS: No CLPU-like lesions were detected under the following conditions: corneal scratch plus lens wear with no bacteria; corneal scratch plus S. epidermidis colonized lenses; corneal scratch without lens wear and with S. aureus applied to eyes; no scratch plus S. aureus colonized lenses; and corneal scratch plus contact lenses colonized by dead S. aureus. CLPU-like lesions were found only when the corneas were scratched and contact lenses colonized by viable S. aureus were applied to the eye. The histology of the lesions demonstrated a frank epithelial break with underlying stromal infiltration. Only low numbers of bacteria could be cultivated from the corneas with CLPU-like lesions. The CLPU-like lesions shared many similarities with CLPU in humans. CONCLUSION: CLPU-like lesions were only produced by S. aureus and not S. epidermidis in the presence of an epithelial abrasion and contact lens. Thus, we hypothesize that to reduce the incidence of CLPU, contact lenses designed to reduce corneal interaction and repel microbial colonization should be produced.

Animals↗

Developmental behavior of gene expression for brown rice thickness under different environments.

The dynamic changes of genetic effects, including main effects, and genotype x environment (GE) interaction effects on brown rice thickness (BRT) across environments were investigated by using the developmental genetic models. Seven cytoplasmic male sterile lines of indica rice (Oryza sativa L.) as females and five restoring lines as males were used in a factorial design to produce grains of F(1)s and F(2)s in two environments (years) for developmental genetic analysis. The results indicate that genetic effects, especially GE interaction effects of triploid endosperm genes, cytoplasm genes, and diploid maternal plant genes were important to the performance of BRT at various filling stages of rice. The BRT was genetically controlled by the net genetic effects of genes expressed at the early and late filling stages (1-7 days and 15-21 days after flowering, respectively). The differences in net genetic effects under different environments for endosperm, cytoplasm, and maternal plant genes were found, and the net GE interaction effects were more important to BRT at the early filling and mature stages of rice. Some net genetic effects, especially for net cytoplasm effects spasmodically expressed, were detected among filling stages. Higher additive and cytoplasm main effects, along with their interaction effects, were found, which would be useful for selection for BRT in breeding programs. The predicated genetic effects at different filling stages show that the parents of V20 and Xieqingzao were better than others for improving BRT of progenies.

Analysis of Variance↗