Biomedical subjects

Q Fang

Publications and source records attributed to Q Fang.

At least 19 recordsLinked to original sources

PubMed2001

On-line incorporation of cloud point extraction to flow injection analysis.

The on-line incorporation of cloud point extraction (CPE) to flow injection analysis (FIA) is demonstrated for the first time. The technical difficulties of inducing the cloud point phenomenon, separating the surfactant-rich phase from the aqueous phase, and detecting trace amounts of analyte(s) in the presence of the highly scattering surfactant medium in an on-line FIA system were resolved by the following: (1) mixing the sample solution containing the analyte(s) and CPE surfactant with an appropriate salting-out agent, (2) using a collection column to entrap the analyte-containing surfactant aggregates, and (3) employing the peroxyoxalate chemiluminescence reaction for the sensitive and selective determination of the analyte(s) in the presence of surfactant micelles. The figures of merit for the determination of coproporphyrin in pretreated urine samples were as follows: precision, 1.1-2.2% (RSD); limit of detection, 2.0 microg/L; and the calibration curve was linear from 46 to 2319 (micro/L (r = 0.9996).

Coproporphyrins↗

PubMed2001

Preparation and characterization of biodegradable copolyester-starch based foams.

Regular (25% amylose) and waxy corn starches blended to various ratios with Eastar Bio Copolyester 14766 (EBC) were extruded into loose-fill foams using a twin screw extruder. Included in this study were two types of corn starch, three levels of EBC content, and three levels of starch moisture content. Waxy starch produced foams with greater radial expansions and lower unit and bulk densities than regular starch. Regular starch foams had lower water solubility indices (WSIs) than waxy starch foams. Foams made of both types of starch possessed similar mechanical properties. No differences were observed in compressibilities and spring indices of either waxy or regular starch foams. Higher levels of EBC addition resulted in less radial expansion and higher unit and bulk densities. Foams made with 10% EBC had higher compressibility than foams containing 25% EBC. Spring indices of single-piece samples and compressibilities and spring indices of bulk samples were not affected by the differences in the level of EBC addition. At 19% and 22% of moisture contents, foams had greater radial expansion than at 25% moisture content. Unit and bulk densities were not affected by variations in moisture content. At 22% moisture content, a lower WSI of 18.5% was obtained. At 22% moisture content, softer foams were produced. At all three levels of moisture content, no differences were detected in the spring indices.

Biocompatible Materials↗

PubMed2001

A histone fold TAF octamer within the yeast TFIID transcriptional coactivator.

Gene activity in a eukaryotic cell is regulated by accessory factors to RNA polymerase II, which include the general transcription factor complex TFIID, composed of TBP and TBP-associated factors (TAFs). Three TAFs that contain histone fold motifs (yTAF17, yTAF60 and yTAF61) are critical for transcriptional regulation in the yeast Saccharomyces cerevisiae and are found in both TFIID and SAGA, a multicomponent histone acetyltransferase transcriptional coactivator. Although these three TAFs were proposed to assemble into a pseudooctamer complex, we find instead that yTAF17, yTAF60 and yTAF61 form a specific TAF octamer complex with a fourth TAF found in TFIID, yTAF48. We have reconstituted this complex in vitro and established that it is an octamer containing two copies each of the four components. Point mutations within the histone folds disrupt the octamer in vitro, and temperature-sensitive mutations in the histone folds can be specifically suppressed by overexpressing the other TAF octamer components in vivo. Our results indicate that the TAF octamer is similar both in stoichiometry and histone fold interactions to the histone octamer component of chromatin.

Alleles↗

PubMed2000

Micelle-mediated extraction and preconcentration of ginsenosides from Chinese herbal medicine.

The feasibility of employing micelle-mediated extraction as an alternative and effective method for the solubilization, purification and/or preconcentration of active ingredients from herbal products is demonstrated for the first time using the root of American ginseng as a model. When compared to methanol and water, an aqueous surfactant solution containing 10% Triton X-100 yielded faster kinetics and higher recovery for the extraction of various ginsenosides. An experimental design approach (uniform design) was demonstrated as a novel and useful method for the optimization of experimental factors involved in the micelle-mediated extraction process. For the preconcentration of ginsenosides prior to chromatographic determination, a salting-out agent (sodium sulfate) was employed to make the efficient cloud point extraction of both hydrophobic and hydrophilic ginsenosides into the surfactant-rich phase possible, as well as to increase the preconcentration factor by reducing the volume of the surfactant-rich phase.

Chromatography, High Pressure Liquid↗

PubMed2000

Sequence of genome segments 1, 2, and 3 of the grass carp reovirus (Genus Aquareovirus, family Reoviridae).

The genome segments 1, 2, and 3 of the grass carp reovirus (GCRV), a tentative species assigned to genus Aquareovirus, family Reoviridae, were sequenced. The respective segments 1, 2, and 3 were 3949, 3877, and 3702 nucleotides long. Conserved motifs 5' (GUUAUUU) and 3' (UUCAUC) were found at the ends of each segment. Each segment contains a single ORF and the negative strand does not permit identification of consistent ORFs. Sequence analysis revealed that VP2 is the viral polymerase, while VP1 might represent the viral guanylyl/methyl transferase (involved in the capping process of RNA transcripts) and VP3 the NTPase/helicase (involved in the transcription and capping of viral RNAs). The highest amino acid identities (26-41%) were found with orthoreovirus proteins. Further genomic characterization should provide insight about the genetic relationships between GCRV, aquareoviruses, and orthoreoviruses. It should also permit to precise the taxonomic status of these different viruses.

Animals↗

PubMed2000

The resonant recognition model (RRM) predicts amino acid residues in highly conserved regions of the hormone prolactin (PRL).

The resonant recognition model (RRM) is a model which treats the protein sequence as a discrete signal. It has been shown previously that certain periodicities (frequencies) in this signal characterise protein biological function. The RRM was employed to determine the characteristic frequencies of the hormone prolactin (PRL), and to identify amino acids ('hot spots') mostly contributing to these frequencies and thus proposed to mostly contribute to the biological function. The predicted 'hot spot' amino acids, Phe-19, Ser-26, Ser-33, Phe-37, Phe-40, Gly-47, Gly-49, Phe-50, Ser-61, Gly-129, Arg-176, Arg-177, Cys-191 and Arg-192 are found in the highly conserved amino-terminal and C-terminus regions of PRL. Our predictions agree with previous experimentally tested residues by site-direct mutagenesis and photoaffinity labelling.

Algorithms↗

PubMed2000

Cartilage-reactive T cells in rheumatoid synovium.

Rheumatoid arthritis (RA) is an inflammatory polyarthritis genetically linked to HLA-DR4 and related haplotypes. RA synovial tissue is characterized by T cell infiltration and activation of macrophage-like cells, strongly implicating a T cell-antigen-presenting cell (APC) interaction in RA pathogenesis. To investigate the nature of the antigens driving the T cell response, synovial tissue was obtained from a patient with chronic RA and T cells were enriched. These T cells were stimulated by endogenous APC from the same synovial tissue. The T cell lines were subsequently evaluated for responsiveness to autologous APC and cartilage antigens. Specific proliferative responses to autologous APC which were enhanced by cartilage extract were seen. Immunomagnetic bead selection and RT-PCR was used to identify TCR alphabeta pairs which appeared to respond to antigen(s) in the cartilage extract. T cell clones derived from the same joint were shown to release IL-2 in response to the cartilage extract and expressed a related TCR. With these experiments we have shown direct evidence that autoreactive T cells are found within the inflamed rheumatoid synovium and, further, that the antigens driving these T cells are cartilage derived. Since the antigens recognized by these populations of T cells are found within cartilage our data provides evidence that RA pathology could be related to a self-driven autoimmune response to cartilage proteins.

Adult↗

PubMed2000

Targeted deletion of the tub mouse obesity gene reveals that tubby is a loss-of-function mutation.

The mouse tubby phenotype is characterized by maturity-onset obesity accompanied by retinal and cochlear degeneration. A positional cloning effort to find the gene responsible for this phenotype led to the identification of tub, a member of a novel gene family of unknown function. A splice defect mutation in the 3' end of the tub gene, predicted to disrupt the C terminus of the Tub protein, has been implicated in the genesis of the tubby phenotype. It is not clear, however, whether the Tub mutant protein retains any biological activity, or perhaps has some dominant function, nor is it established that the tubby mutation is itself responsible for all of the observed tubby phenotypes. To address these questions, we generated tub-deficient mice and compared their phenotype to that of tubby mice. Our results demonstrate that tubby is a loss-of-function mutation of the tub gene and that loss of the tub gene is sufficient to give rise to the full spectrum of tubby phenotypes. We also demonstrate that loss of photoreceptors in the retina of tubby and tub-deficient mice occurs by apoptosis. In addition, we show that Tub protein expression is not significantly altered in the ob, db, or melanocortin 4 receptor-deficient mouse model of obesity.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Establishment and chimera analysis of 129/SvEv- and C57BL/6-derived mouse embryonic stem cell lines.

Hundreds of new mutant mouse lines are being produced annually using gene targeting and gene trap approaches in embryonic stem (ES) cells, and the number is expected to continue to grow as the human and mouse genome projects progress. The availability of robust ES cell lines and a simple technology for making chimeras is more attractive now than ever before. We established several new ES cell lines from 129/SvEv and C57BL/6 mice and tested their ability to contribute to the germline following blastocyst injections and/or the less expensive and easier method of morula-ES cell aggregation. Using morula aggregation to produce chimeras, five newly derived 129/SvEv and two C57BL/6 ES cell lines tested at early passages were found to contribute extensively to chimeras and produce germline-transmitting male chimeras. Furthermore, the two 129S/vEv ES cell lines that were tested and one of the C57BL/6 ES cell lines were able to maintain these characteristics after many passages in vitro. Our results indicate that the ability of ES cells to contribute strongly to chimeras following aggregation with outbred embryos is a general property of early passage ES cells and can be maintained for many passages. C56BL/6-derived ES cell lines, however, have a greater tendency than 129-derived ES cell lines to lose their ability to colonize the germline.

Animals↗

PubMed2000

[Analysis of chromosomal karyotypes in 300 fetal blood samples during the second and third trimesters of gestation].

OBJECTIVE: To analyze the fetal chromosomal karyotypes from the blood samples obtained by cordocenteses during the second and third trimesters, and to investigate the types of chromosomal abnormalities, as well as the relationship between the abnormal karyotypes and the indications of prenatal diagnosis. METHODS: Cordocenteses were performed in 300 pregnant women with different indications for prenatal diagnosis during the 18 to 38 gestational weeks, and fetal chromosomal karyotypes were examined. RESULTS: Twenty three chromosomal abnormalities(7.7%) were checked out. In the second trimester, there were 15 abnormalities in 174 samples(8. 6%); whereas in the third trimester, it was 8 out of 126(6.3%), P=0. 77. Trisomy, the leading abnormality, consisted of 60.9%(14/23) of all abnormalities and 9 out of 14 were trisomy 21, which was 39. 1%(9/23). In those aged over 35 years, trisomy 21 was detected in 5 of 92(5.4%), and in the age under 35 years, it was 4 out of 208(1. 9%), P=0.26. Thirty three women had the history of giving a birth of trisomy 21 previously, this time, however, no one was recurrent. Highest chromosomal aberration rate, 26.3%(5/19), was detected in the fetuses with intrauterine growth retardation(IUGR), and all were trisomy. Balanced translocation was found in 5 fetuses (1 associated with Robertsonian translocation), which was 21.7%(5/23). CONCLUSION: During the second and third trimesters, the rate of chromosomal abnormality is 7.7% in those fetuses who have maternal indications for prenatal diagnosis. trisomy, especially trisomy 21, is the most common abnormal karyotype found in these periods and in advanced maternal age, as well as in severe IUGR.

Adult↗

PubMed2000

[Differentiation of achondroplasia and other similar genetic dwarfism by FGFR3 gene analysis].

OBJECTIVE: To study the gene mutation of Chinese patients with achondroplasia(ACH) and to set up a simple and rapid molecular diagnostic method to differentiate ACH from other similar genetic dwarfism. METHODS: The specific fragment of fibroblast growth factor receptor 3(FGFR3) transmembrane domain was amplified from dried blood spots of 21 patients with ACH and 6 suspicious patients with ACH by polymerase chain reaction, then mutation was screened and detected by restrictive enzyme analysis, single strand conformation polymorphism(SSCP) and denaturing gradient gel electrophoresis(DGGE). RESULTS: One out of 6 suspicious cases was ACH and 5 were pseudoachondroplasia(PSACH). Twenty-one out of 22 patients with ACH bore a G to A transition at nucleotide 1138 and 1 bore a G to C transversion at this same position. CONCLUSION: The nucleotide 1138 of FGFR3 gene is also the hotspot of mutation in Chinese patients with ACH. A simple and rapid molecular diagnostic method has been set up to differentiate ACH from other similar genetic dwarfism.

Achondroplasia↗

PubMed1999

Influence of fetal to neonatal transition on nitric oxide synthase expression in the nucleus tractus solitarius in sheep.

Transition from fetal to newborn life is accompanied by a marked rise in circulating norepinephrine (NE) concentrations though arterial blood pressure does not substantively change. Nitric oxide (NO) plays an important role in the central regulation of sympathetic tone in the nucleus tractus solitarius (NTS) and neuronal NO synthase (nNOS) expression is functionally regulated in the brain. The purpose of these studies was to determine the influence of transition at birth on nNOS expression in the brainstem nuclei, particularly in the NTS, associated with changes in arterial pressure and plasma NE concentration. Experiments were performed using time-dated gestational ewes with twin fetuses. Arterial blood pressure was recorded and arterial blood NE concentrations were measured in the term fetus (gestational 147-148 days) and newborn lambs (4 h of postnatal age). The fetal and newborn animals were then perfused with 4% paraformaldehyde. Sections of the medulla were examined by using both immunolabeling with a polyclonal antibody directed against nNOS and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry, a marker for expression of nNOS. Micrographs were quantified using a microscope with reticule grid to measure the number of positive cells containing color staining in the brainstem nuclei. Plasma NE concentration in the newborn was more than two-fold greater compared to fetal values but mean arterial blood pressure was similar between fetus and newborn. The nNOS positive cells and NADPHd positive cells were significantly increased in the medial NTS (mNTS) of the newborn compared to fetus. nNOS immunoreactivity and NADPHd reactivity tended to increase in the rostral ventral medulla (RVM) in newborn, but were not altered in other brainstem nuclei during the transition from fetal to newborn life. The results suggest that nNOS expression in the mNTS is predominately enhanced at 4 h of neonatal age vs. the term fetus. We conclude that elevated circulating NE is associated with up-regulation of nNOS in the mNTS which may serve a protective role in central regulation of neonatal arterial blood pressure.

Animals↗

PubMed1999

Ligation of CD28 in vivo induces CD40 ligand expression and promotes B cell survival.

Functional activation of T cells requires ligation of Ag receptors with specific peptides presented by MHC molecules on APCs concurrent with appropriate contacts of cell surface accessory molecules. Among these accessory molecules, interactions between CD28/CTLA-4 with B7 family members (CD80 and CD86) and CD40 with CD40 ligand (CD40L) play a decisive role in regulating the progression of balanced immune responses. However, most information regarding the role of accessory molecules in immune responses has been derived in the context of signals from the TCRs. Little understanding has been achieved regarding the consequence of ligation of costimulation molecules in absence of signals from the TCR. By employing an in vivo murine system, we show, herein, that ligation of CD28 alone with anti-CD28 Abs leads to a dramatic enlargement of the peripheral lymphoid organs characterized primarily by the expansion of B cells. B cells from anti-CD28-treated mice are resistant to spontaneous and anti-IgM-induced apoptosis. These cells are also unsusceptible to FasL-mediated apoptosis. Interestingly, this in vivo effect of CD28 on B cells is largely mediated by inducing the expression of CD40L, since coadministration of a blocking Ab against CD40L inhibited CD28-mediated B cell survival and expansion. Therefore, CD28-mediated expression of CD40L may play an important role in the regulation of lymphocyte homeostasis.

Animals↗

PubMed1999

Boswellic acid acetate induces differentiation and apoptosis in leukemia cell lines.

Boswellic acid acetate (BC-4), a compound isolated from the herb Boswellia carterii Birdw., can induce differentiation and apoptosis of leukemia cells. Based on cell morphology and NBT reduction, BC-4 induced monocytic differentiation of myeloid leukemia HL-60, U937 and ML-1 cells at a dose under 12.5 microg/ml (24.2 microM). BC-4 was a potent inducer, with 90% of the cells showing morphologic changes and 80-90% of the cells showing NBT reduction. Specific and non-specific esterase were also increased by BC-4. Based on benzidine staining assay, BC-4 failed to induce erythroid leukemia DS-19 and K562 cells differentiation. In contrast to its selective differentiation effect, BC-4 strongly inhibited growth of all cell lines tested. The growth inhibition effect was dose- and time-dependent. In HL-60 cells, 20 microg/ml (38.8 microM) of BC-4 decreased viable cell number by 60% at 24 h, whereas at 3 days there was virtually no viable cells. Morphologic and DNA fragmentation analysis proved that BC-4 induced cell apoptosis. The dual apoptotic and differentiation effects of BC-4 suggest that it may be a powerful agent in the treatment of leukemia.

Antineoplastic Agents, Phytogenic↗

PubMed1998

Targeted mutation reveals a central role for SR-BI in hepatic selective uptake of high density lipoprotein cholesterol.

Scavenger receptor BI (SR-BI) is a cell surface receptor that binds high density lipoproteins (HDL) and mediates selective uptake of HDL cholesteryl esters (CE) in transfected cells. To address the physiological role of SR-BI in HDL cholesterol homeostasis, mice were generated bearing an SR-BI promoter mutation that resulted in decreased expression of the receptor in homozygous mutant (designated SR-BI att) mice. Hepatic expression of the receptor was reduced by 53% with a corresponding increase in total plasma cholesterol levels of 50-70% in SR-BI att mice, attributable almost exclusively to elevated plasma HDL. In addition to increased HDL-CE, HDL phospholipids and apo A-1 levels were elevated, and there was an increase in HDL particle size in mutant mice. Metabolic studies using HDL bearing nondegradable radiolabels in both the protein and lipid components demonstrated that reducing hepatic SR-BI expression by half was associated with a decrease of 47% in selective uptake of CE by the liver, and a corresponding reduction of 53% in selective removal of HDL-CE from plasma. Taken together, these findings strongly support a pivotal role for hepatic SR-BI expression in regulating plasma HDL levels and indicate that SR-BI is the major molecule mediating selective CE uptake by the liver. The inverse correlation between plasma HDL levels and atherosclerosis further suggests that SR-BI may influence the development of coronary artery disease.

Animals↗

PubMed1998

Construction and binding kinetics of a soluble granulocyte-macrophage colony-stimulating factor receptor alpha-chain-Fc fusion protein.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) activity is mediated by a cellular receptor (GM-CSFR) that is comprised of an alpha-chain (GM-CSFRalpha), which specifically binds GM-CSF, and a beta-chain (betac), shared with the interleukin-3 and interleukin-5 receptors. GM-CSFRalpha exists in both a transmembrane (tmGM-CSFRalpha) and a soluble form (sGM-CSFRalpha). We designed an sGM-CSFRalpha-Fc fusion protein to study GM-CSF interactions with the GM-CSFRalpha. The construct was prepared by fusing the coding region of the sGM-CSFRalpha with the CH2-CH3 regions of murine IgG2a. Purified sGM-CSFRalpha-Fc ran as a monomer of 60 kDa on reducing SDS-polyacrylamide gel electrophoresis but formed a trimer of 160-200 kDa under nonreducing conditions. The sGM-CSFRalpha-Fc bound specifically to GM-CSF as demonstrated by standard and competitive immunoassays, as well as by radioligand assay with 125I-GM-CSF. The sGM-CSFRalpha-Fc also inhibited GM-CSF-dependent cell growth and therein is a functional antagonist. Kinetics of sGM-CSFRalpha-Fc binding to GM-CSF were evaluated using an IAsys biosensor (Affinity Sensors, Paramus, NJ) with two assay systems. In the first, the sGM-CSFRalpha-Fc was bound to immobilized staphylococcal protein A on the biosensor surface, and binding kinetics of GM-CSF in solution were determined. This revealed a rapid koff of 2.43 x 10(-2)/s. A second set of experiments was performed with GM-CSF immobilized to the sensor surface and the sGM-CSFRalpha-Fc in solution. The dissociation rate constant (koff) for the sGM-CSFRalpha-Fc trimer from GM-CSF was 1.57 x 10(-3)/s, attributable to the higher avidity of binding in this assay. These data indicate rapid dissociation of GM-CSF from the sGM-CSFRalpha-Fc and suggest that in vivo, sGM-CSFRalpha may need to be present in the local environment of a responsive cell to exert its antagonist activity.

Animals↗

PubMed1998

Flow injection on-line photochemical reaction coupled to spectrofluorimetry for the determination of thiamine in pharmaceuticals and serum.

The photochemical reaction of thiamine was studied with a photochemical reactor made by coiling a knotted PTFE reactor around a low-pressure mercury lamp. Acetone, which was previously reported to be a sensitizer for the photochemical reaction that took place in situ in a flow-through cell, severely depressed the fluorescence signal of the photochemical reaction that took place on-line in the knotted PTFE reactor when sodium sulfite was involved in the photochemical reaction. Experiments revealed that the effect of acetone on the photochemical reaction was dependent on the intensity of the irradiation that was used to induce the photochemical reaction, and that acetone might impair the photochemically induced fluorescence if strong UV irradiation was applied to induce the photochemical reaction and sodium sulfite was used to enhance the fluorescence signal. Based on these observations, a flow injection on-line photochemical-spectrofluorimetric method for the determination of thiamine was developed without using acetone. With the proposed method, a detection limit of 0.11 micrograms l-1 thiamine, a relative standard deviation of 0.36% for 11 determinations of 1 mg l-1 thiamine and a sampling frequency of 100 h-1 were achieved. The developed method was successfully applied to the determination of the thiamine content in various pharmaceutical preparations and serum.

Flow Injection Analysis↗

PubMed1998

Structural motifs in rheumatoid T-cell receptors.

The linkage of rheumatoid arthritis (RA) to HLA-DR haplotypes, high levels of HLA-DR expression, and T-cell infiltration in the joints, indicate a central role for the interaction of T-cell receptors (TCR) with antigen (Ag) + major histocompatibility complex (MHC) complexes in pathogenesis. Receptor analysis in RA has uncovered a restricted heterogeneity of TCR transcripts, suggesting an antigen-driven response. We analyzed the sequence and structural features of RA-associated TCRs in light of the recently published TCR crystal structures. The surface-exposed residues of the third complementarity-determining region (CDR3s) showed preferential use of certain amino acid residues when sequences derived from synovial fluid or tissue were compared with those derived from peripheral blood, particularly for alpha chains. Sequence alignment of oligoclonal synovial TCR CDR3s revealed groupings with similar CDR3 lengths and amino acid compositions, which suggests shared antigen recognition. Given the limitations of analyzing TCR sequences without knowing their structures, we developed several in vivo-activated synovial-tissue Vbeta17 + RA T-cell clones. Two Vbeta17/V alpha7 clones with different CDR3 sequences were analyzed by molecular modeling. Although distinct topologic features were seen, a central patch of residues with similar chemical and geometric characteristics was present in both. Electrostatic maps revealed similar binding surfaces of both alpha domains and central patches, with differences in the beta domains. This suggests that an alpha-domain-focused binding trajectory would allow shared antigen recognition by these TCRs. These studies support recognition of a limited diversity of Ag + MHC complexes by synovial RA TCRs.

Amino Acid Sequence↗