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Biomedical subjects

Q Wu

Publications and source records attributed to Q Wu.

At least 73 records · Page 4Linked to original sources

Gene targeting in hemostasis. Hepsin.

Hepsin is a type II transmembrane serine protease abundantly expressed on the surface of hepatocytes. Biochemical studies have shown that hepsin is an enzyme of 51 kDa with the trypsin-like substrate specificity. Several in vitro studies have suggested that hepsin may play a role in blood coagulation, hepatocyte growth, and fertilization. To determine the functional importance of hepsin, hepsin-deficient mice were generated by homologous recombination. Homozygous hepsin-/- mice were viable and fertile, and grew normally. When analyzed in hemostasis assays, such as tail bleeding time and plasma clotting times, and in vivo modes, such as disseminated intravascular coagulation, septic shock, and acute liver regeneration, hepsin-/- mice had similar phenotypes as wild-type controls. Liver weight and serum concentrations of liver-derived proteins or enzymes were also similar in hepsin-/- and wild-type mice. No abnormalities were identified in major organs in hepsin-/- mice in histological examinations. These results indicate that hepsin is not an essential enzyme for normal hemostasis, embryogenesis, and maintenance of normal liver function. Unexpectedly, serum concentrations of bone-derived alkaline phosphatase were approximately two-fold higher in both male and female hepsin-/- mice than those in wild-type controls. The underlying mechanism for this phenotype and long-term effects of hepsin deficiency remain to be determined.

Animals↗

Signal via lymphotoxin-beta R on bone marrow stromal cells is required for an early checkpoint of NK cell development.

NK cells play an important role in the immune system but the cellular and molecular requirements for their early development are poorly understood. Lymphotoxin-alpha (LTalpha)(-/-) and LTbetaR(-/-) mice show a severe systemic reduction of NK cells, which provides an excellent model to study NK cell development. In this study, we show that the bone marrow (BM) or fetal liver cells from LTalpha(-/-) or LTbetaR(-/-) mice efficiently develop into mature NK cells in the presence of stromal cells from wild-type mice but not from LTalpha(-/-) or LTbetaR(-/-) mice. Direct activation of LTbetaR-expressing BM stromal cells is shown to promote to early NK cell development in vitro. Furthermore, the blockade of the interaction between LT and LTbetaR in adult wild-type mice by administration of LTbetaR-Ig impairs the development of NK cells in vivo. Together, these results indicate that the signal via LTbetaR on BM stromal cells by membrane LT is an important pathway for early NK cell development.

Adoptive Transfer↗

Complementary effects of TNF and lymphotoxin on the formation of germinal center and follicular dendritic cells.

The formation of germinal centers (GC) around follicular dendritic cells (FDC) is a critical step in the humoral immune responses that depends on the cooperative effects of B cells and T cells. Mice deficient in either TNF or lymphotoxin (LT) fail to form both GC and FDC network in B cell follicles. To test a potential complementary effect of TNF and LT, a mixture of bone marrow cells from TNF(-/-) mice and LT alpha(-/-) mice was transferred into irradiated LT alpha(-/-) mice or TNF(-/-) mice. Interestingly, the formation of both GC and FDC clusters in B cell follicles was restored in such chimeric mice, suggesting that TNF and LT from different cells could complement one another. To identify the exact contributions of each subset to the complementary effect of TNF and LT, different sources of T and B cells from LT alpha(-/-) mice or TNF(-/-) mice were used for reconstitution. Our study demonstrates that either T or B cell-derived TNF is sufficient to restore FDC/GC in the presence of LT-expressing B cells. However, TNF itself is not required for GC reactions if the FDC network is already intact. Thus, the development and maintenance of these lymphoid structures depend on a delicate interaction between TNF and LT from different subsets of lymphocytes.

Adjuvants, Immunologic↗

Is the SLC7A10 gene on chromosome 19 a candidate locus for cystinuria?

One of the genes (SLC7A9) that causes cystinuria, an inborn error of amino acid transport, is localized to 19q13. Close examination of human genomic DNA sequences has identified a similar gene (SLC7A10) that also maps to the 19q13.1 region and is highly expressed in kidney. The homologies between SLC7A9 and SLC7A10 are likely the result of gene duplication. SLC7A10 is known to encode a protein with a function similar to that of the SLC7A9 gene product. To determine if mutations in the SLC7A10 gene could also cause cystinuria, we characterized the primary genomic structure and sequenced the 11 exons and surrounding sequences from 10 unrelated patients with cystinuria. We identified one missense mutation which may account for cystinuria in one family. We also observed one intronic change, as well as one silent mutation, that were seen only in cystinuria patients. We therefore suggest that the SLC7A10 gene warrants further investigation as another candidate gene for cystinuria.

Amino Acid Sequence↗

Studies on the frequencies of vitamin D receptor gene polymorphism in postmenopausal women of Han and Kazak nationality in China.

Polymorphyism of the vitamin D receptor (VDR) gene is reported to play a major role in variations of the genetic regulation of bone mass. The role of VDR polymorphisms within various ethnic populations is also undetermined. The purpose of the present study was to determine the frequencies of VDR genotypes in the Han and Kazak nationalities in China. We analyzed the polymorphism defined by the Bsm1 restriction enzyme using polymerase chain reaction-restriction fragment length polymorphism in 179 healthy postmenopausal Han women and 63 healthy postmenopausal Kazak women. The genotype frequencies of the VDR were calculated later using the Hardy-Weinberg equilibrium formula. The results showed that for women of Han nationality, the bb, Bb, and BB genotypes accounted for 90.5%, 9.5%, and 0%, respectively. In Kazak women, the respective frequencies were 38.1%, 55.56%, and 6.35%. We found that there was a significant difference between women of Han and Kazak nationality in terms of the frequency of distribution of VDR genotype (P < 0.001). There was a similar distribution of VDR genotypes in Kazak women to that seen for the Caucasian population of the USA. The results of the present study provide further evidence on the different pathogeny of osteoporosis in various ethnic groups.

Bone Density↗

A new operation for ventricular septal defect with aortic incompetence.

Two boys, 10 years of age, with ventricular septal defect and severe aortic insufficiency were treated using a new method. An autograft of anterior pulmonary artery wall and the adjacent leaflet and subpulmonary muscle was excised. The excised pulmonary artery segment was used to replace the prolapsed right coronary cusp, and the autograft muscle was used to close the ventricular septal defect. The right ventricular outflow tract and pulmonary valve were repaired using an autologous pericardial patch with an autologous single cusp. The results in both cases were satisfactory.

Aortic Valve Insufficiency↗

Repair of atrial septal defect through a minimal right vertical infra-axillary thoracotomy in a beating heart.

We present a minimally invasive and cosmetic approach to repair atrial septal defect. Fifty-five atrial septal defects were successfully repaired through a minimal right vertical infra-axillary thoracotomy. Every repair was performed on the beating heart with continuous suction through aortic needle vents and with the head of the patient lowered. This approach can be used not only in adults but also in young girls.

Axilla↗

Purification and characterization of White Spot syndrome virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii.

Penaeid shrimp is the natural host of White Spot Syndrome Virus (WSSV) that can cause high mortality in the infected hosts. Attempts to obtain sufficient amounts of purified intact WSSV for characterization have been unsuccessful. Using crayfish, Cambarus clarkii as a proliferation system, a large amount of infectious WSSV was reproduced and intact WSSV viral particles were purified with a new isolation medium by ultra-centrifugation. Purified WSSV particles were very sensitive to organic solvents and the detergent, Triton X-100. The size of the rod-shape, somewhat elliptical, intact WSSV was 110-130x260-350 nm with a long, tail-like envelope extension. The naked viral nucleocapsid was about 80x350 nm, and it possessed 15 spiral and cylindrical helices composed of 14 globular capsomers along its long axis, and a 'ring' structure at one terminus. Distinct WSSV genome DNA patterns were obtained when the purified genomic dsDNA of WSSV was digested with five different restriction enzymes (HindIII, XhoI, BamHI, SalI, and SacI). In addition, at least 13 major and distinct protein bands could be observed when purified intact WSSV viruses were separated by SDS-PAGE followed by Coomassie Brilliant R-250 staining. The estimated molecular weights of these proteins were 190, 84, 75, 69, 68, 58, 52, 44, 28, 27.5, 23, 19, and 16 kD, respectively. Both the 44 and 190 kD proteins were easily removed if the hemolymph from the WSSV infected crayfish was transiently treated with 1% Triton X-100 before it was subjected to gradient centrifugation, indicating that both of them are located on the surface of the viral envelope. These characteristics are consistent with WSSV isolated from the penaeid shrimp.

Animals↗

Dosimetric validation for multileaf collimator-based intensity-modulated radiotherapy: a review.

The creation of intricate dose distributions produced by intensity-modulated radiotherapy (IMRT) depends on complex planning systems and specialized mechanical devices. The many possible sources of inaccuracy and the complexity of the dose maps themselves require that a substantial effort be made to ensure that calculated and delivered dose distributions agree. This review provides an overview of the current status of the validation of dose predictions of IMRT planning systems by comparisons with measurements. Emphasis is placed on multileaf collimator- (MLC) based IMRT. Discrepancies between calculations and measurements may be due to any of 3 causes: errors and uncertainties in the dose calculation algorithm, in measurements, or in beam delivery by the accelerator/MLC combination. Some of the factors affecting dosimetry include: the technique employed for modulating the fluence, the dose calculation algorithm and other aspects of the planning system, mechanical limitations of the MLC hardware, dosimetric characteristics of the MLC, such as MLC leakage and rounded leaf ends, the choice of dosimeter, and the measurement geometry and technique. The advantages and drawbacks of various dosimeters including film, ion chambers, thermoluminescent dosimetry, and electronic portal imaging devices are discussed. The steps involved in validating dosimetrically a planning system are outlined, including the various fields that need to be measured, the phantoms that may be used, and measurement techniques. The achievable accuracy of dosimetry for IMRT is discussed.

Radiometry↗

Pyrolytic characteristics of microalgae as renewable energy source determined by thermogravimetric analysis.

Two kinds of autotrophic microalgae, Spirulina platensis (SP) and Chlorella protothecoides (CP) were pyrolyzed at the heating rates of 15, 40, 60 and 80 degrees C/min up to 800 degrees C in a thermogravimetric analyzer to investigate their pyrolytic characteristics. Three stages (dehydration, devolatilization and solid decomposition) appeared in the pyrolysis process. SP and CP mainly devolatilized at 190-560 degrees C and 150-540 degrees C, respectively. A total volatile yield of about 71% was achieved from each microalga. As the heating rate increased, a lateral shift to higher temperatures was observed in their thermograms, and the instantaneous maximum and average reaction rates in the devolatilization stage were increased while the activation energy was decreased. The value of activation energy for CP pyrolysis was 4.22-5.25 x 10(4), lower than that of SP (7.62-9.70 x 10(4)), and the char in final residue of CP was 14.00-15.14%, less than that of SP by 2-3%. This indicated that CP is preferable for pyrolysis over SP. The experimental results may provide useful data for the design of pyrolytic processing systems using planktonic microalgae as feedstock.

Bioelectric Energy Sources↗

Fourier-transform Raman and infrared spectroscopic analysis of dipyrrinones and mesobilirubins.

The Fourier-transform Raman (FT-Raman), infrared (FT-IR), and UV-visible absorption spectra of four dipyrrinones and two mesobilirubins have been investigated in the solid state and in CH2Cl2 solutions. A detailed spectral analysis, assignment and discussion of these spectra are presented. The bands at 1735-1738, 1691-1707 and 1359-1377 cm(-1) which were assigned to the stretching vibrations of the C-O-C and C-O-H and symmetric deformation of C-H bonds, respectively, can act as a marker to distinguish the compounds of this class. The striking differences between the spectra of the compounds suggest that mesobilirubin XIIIalpha is tending to adopt as ridge-tile conformation, rather than linear conformation.

Bilirubin↗

Production of poly-3-hydroxybutyrate by Bacillus sp. JMa5 cultivated in molasses media.

A strain of Bacillus sp. coded JMa5 was isolated from molasses contaminated soil. The strain was able to grow at a temperature as high as 45 degrees C and in 250 g/l molasses although the optimal growth temperature was 35-37 degrees C. Cell density reached 30 g/l 8 h after inoculation in a batch culture with an initial concentration of 210 g/l molasses. Under fed-batch conditions, the cells grew to a dry weight of 70 g/l after 30 h of fermentation. The strain accumulated 25-35%, (w/w) polyhydroxybutyrate (PHB) during fermentation. PHB accumulation was a growth-associated process. Factors that normally promote PHB production include high ratios of carbon to nitrogen, and carbon to phosphorus in growth media. Low dissolved oxygen supply resulted in sporulation, which reduced PHB contents and dry weights of the cells. It seems that sporulation induced by reduced supply of nutrients is the reason that PHB content is generally low in the Bacillus strain.

3-Hydroxybutyric Acid↗

Molecular cloning and characterization of a novel Dehydrogenase/reductase (SDR family) member 1 genea from human fetal brain.

Short-chain dehydrogenases/reductases (SDR) constitute a large protein family of NAD(P)(H)-dependent oxidoreductase. They are defined by distinct, common sequence motifs and show a wide range of substrate specialisms. By large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a novel human SDR-type dehydrogenase/reductase gene named Dehydrogenase/reductase (SDR family) member 1 (DHRS1). The DHRS1 cDNA is 1411 base pair in length, encoding a 314-amino-acid polypeptide which has a SDR motif. Northern blot reveals two bands, of about 0.9 and 1.4 kb in size. These two forms are expressed in many tissues. The DHRS1 gene is localized on chromosome 14q21.3. It has 9 exons and spans 9.2 kb of the genomic DNA.

Amino Acid Sequence↗

Approaches to the remediation of a polychlorinated biphenyl (PCB) contaminated soil--a laboratory study.

A soil that had been historically contaminated with Aroclor 1242, 1248, 1254 and 1260 was decontaminated by two surfactant-mediated cleaning procedures that had been chosen to mimic ex-situ washing and in-situ soil flushing processes. A preliminary screening selected four surfactants (from 17 commercial formulations) for their ability to mobilise PCBs from the soil while suffering minimal losses to the supercritical carbon dioxide (scCO2) that was used in a separate back-extraction procedure. The mobilisation was enhanced, with minimal foam formation, by the presence of 17% (v/v) IBMK in the surfactant suspension. Each of the four surfactants, at 1, 3, or 5% (v/v) concentration, was evaluated by (i) 15 successive 10 min sonication-filtrations and (ii) continuous soil column flushing during 20 h. Each filtrate from (i) and samples, taken at hourly intervals, from (ii) were analysed for their PCB and surfactant content. Both extraction procedures mobilised PCBs efficiently when extended for longer periods and were modelled accurately as the sum of a constant and single-term exponential increase to a maximum. The predicted number of replicate stages required to mobilise 50% of the toxicants (t50) varied from 7 to 3 for sonication-washing of the soil (10 g) or from 6.8 to 2.8 h for column flushing of 30 g soil and decreased as the concentration of surfactant in the aqueous phase was increased. The combined PCB-laden aqueous suspensions were then back-extracted efficiently with scCO2 and the eluate was dechlorinated quantitatively as it traversed a short, heated column of silver-iron bimetallic mixture.

Carbon Dioxide↗

Extractions of polychlorinated biphenyl (PCB) compounds from surfactant suspension/soil extracts with dechlorination on-line.

Polychlorinated biphenyl (PCB) formulation (Aroclor 1242 or 1248)-surfactant (Brij 97 or Triton CF54) emulsions (0.1-1.0%, v/v) were extracted with supercritical carbon dioxide (scCO2). The declinations curve that resulted for each loading of PCB substrate was accurately modelled as a single-exponential decay, and the half-life [t1/2, 10.76 min +/- 2% (Brij 97) or 10.0 min +/- 14% (Triton CF54)] was independent of the level of loading. When the extraction was combined with on-line dechlorination, t1/2 was increased to 17.62 min +/- 6% or 16.46 min +/- 5%, respectively The dechlorination reactor contributed appreciably to the back-pressure of the system so that, under identical scCO2 head pressures, the flow rate at the exit of the system was reduced from 1500 mL min-1 (as decompressed gas) to 900 mL min-1. When corrected for the difference in flow rates, the declination curves in the absence and presence of the reactor became virtually identical. For a surfactant suspension from a soil that was field-burdened with only 6 ppm PCBs, extended operation (extraction-dechlorination) for 15 h, during which the substrate suspension was replaced every 30 min, caused no loss of the dechlorination efficiency; however, for 1% (v/v) Aroclor 1242 or 1248 in surfactant suspension, the reactor gradually lost reactivity over 5 h of continued operation. However, the reactivity could be restored virtually completely by purging the reactor column with scCO2 for 3 h or washing the column at ambient temperature with 30 mL of methanol-water (7 + 3, v/v). Mass spectrometry of the reaction products indicated that the principal products included biphenyl and toluene, but methylated benzenes and phenols were also present.

Chemistry Techniques, Analytical↗

A novel fatty acid response element controls the expression of the flight muscle FABP gene of the desert locust, Schistocerca gregaria.

In many tissues, fatty acid binding protein (FABP) expression is stimulated by exposure to elevated fatty acid levels. In contrast to the FABP genes expressed in other tissues, the molecular mechanisms that mediate the upregulation of the muscle FABP gene have not been elucidated. We have studied the expression of locust flight muscle FABP, a protein that is highly homologous to the mammalian H-FABPs. A 130-bp promoter fragment of the locust gene, which includes a canonical TATA box and several GC boxes, is sufficient for the transcription of a reporter gene in mammalian L6 myoblasts. Twofold higher expression rates are observed when the promoter contains 280 bp or more of upstream sequence. Treatment of myoblasts with various fatty acids leads to a marked increase of expression in the longer constructs, but not in the minimal promoter. We have identified a 19-bp inverted repeat (-162/-180) as the element responsible for the fatty acid-mediated induction of gene expression. Deletion of this element eliminates the fatty acid response, and gel shift analysis demonstrates specific binding to nuclear proteins from both L6 myoblasts and locust flight muscle cells. This fatty acid response element bears no similarity to any known transcription factor binding site. A similar palindrome was also found in the promoter of the Drosophila melanogaster muscle FABP gene, and in reverse orientation upstream of all mammalian heart FABP genes. Given the structural and functional conservation of muscle FABPs and their genes, it is possible that this fatty acid response element also modulates the expression of the mammalian H-FABP genes.

Animals↗

Comparative analysis of polychlorinated biphenyl-dechlorinating communities in enrichment cultures using three different molecular screening techniques.

The catalysts for many microbially mediated environmental processes such as the dechlorination of polychlorinated biphenyls (PCBs) have been difficult to identify by traditional isolation techniques. Numerous, as yet unsuccessful, attempts have been made to isolate and culture the dechlorinating species. To overcome this limitation, amplified rDNA restriction analysis (ARDRA) of a clone library, denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (TRFLP) were used concurrently to compare their effectiveness for characterizing an enriched microbial community. These methods were applied to enrichment cultures that selectively dechlorinated double-flanked chlorines in the PCB congener 2,3,4,5 chlorinated biphenyl. The methods have different biases, which were apparent from discrepancies in the relative clone frequencies (ARDRA), band intensities (DGGE) or peak heights (TRFLP) from the same enrichment culture. However, each method was effectively qualitative and identified the same organisms: a low G + C Gram-positive eubacterium, an organism most similar to the green non-sulphur bacteria, an Aminobacterium sp. and a Desulfovibrio sp. Overall, in community fingerprinting and preliminary identification, DGGE proved to be the most rapid and effective tool for the monitoring of microorganisms within a highly enriched culture. TRFLP results corroborated DGGE fingerprint analysis; however, identification required the additional step of creating a clone library. ARDRA provided an in-depth analysis of the community and this technique detected slight intraspecies sequence variation in 16S rDNA. These molecular methods are common in environmental microbiology, but rarely are they compared with the same sample site or culture. In general, all three methods detected similar community profiles, but inherent biases resulted in different detection limits for individual OTUs (operational taxonomic units).

Ampicillin↗