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Biomedical subjects

M Yang

Publications and source records attributed to M Yang.

At least 55 records · Page 3Linked to original sources

Development of defensive behavior and conditioning to cat odor in the rat.

Laboratory rats show a range of defensive behaviors, including freezing, avoidance, and risk assessment upon exposure to cat odor, an unconditioned but highly effective threat stimulus. This study examined defensive behaviors, and the rapid conditioning to context plus cue, of these behaviors, in 18-, 26-, and 38-day-old male and female rats exposed to cat odor. Rats were placed individually in a runway with a cloth covered (control or saturated with cat fur/skin odor) block for a 10-min trial. On the following day, a similar trial involved an odorless block. On the odor exposure day, rats of all ages showed less contact with the odor block than with the control block. The 26- and 38-day-old rats, but not the 18-day-old rats, also showed locomotor suppression, more avoidance of the area where the odor block was located, and more risk assessment than no-odor controls. On a test of conditioned behavior 24 h following exposure, 26- and 38-day-old rats exhibited defensive behavior including avoidance and reduction of locomotion while 18-day-old pups did not.

Age Factors↗

Efficient inhibition of multidrug-resistant human tumors with a recombinant bispecific anti-P-glycoprotein x anti-CD3 diabody.

Overexpressing of P-glycoprotein (Pgp) has been shown to be responsible for cancer resistance to multiple chemotherapeutic agents. Immunotherapy with biological agents, such as bispecific antibodies (BsAbs), may represent a promising approach to overcome the emergence of drug resistance. Here we constructed a recombinant BsAb, a diabody, with specificities to both CD3 on human T-lymphocyte and Pgp on cancer cells. The diabody was produced in Escherichia coli in a soluble functional form and purified by an affinity chromatography with a yield of >4 mg/l culture medium in shaker flask. The diabody binds to both CD3 on T-lymphocytes and Pgp on multidrug-resistant (MDR) tumor cells with affinities that are comparable to its respective parental single chain Fv molecules. In the presence of activated human peripheral blood lymphocytes (PBLs), the diabody mediates effectively the lysis of the Pgp-overexpressing human leukemia K562/A02 and epidermoid carcinoma KBv(200) cells, but is much less potent in mediating the lysis of the parent K562 and KB cells. Further, the diabody localized selectively within the K562/A02 xenografts in mice. When combined with activated PBL, the diabody significantly inhibited the growth of K562/A02 and KBv(200), but had no effect on K562 and KB xenografts. In contrast, treatment with doxorubicin, a standard chemotherapeutic agent, only inhibited the growth of K562 and KB, but had no effect on K562/A02 and KBv(200) xenografts. Taken together, our results suggest that the anti-Pgp x anti-CD3 diabody may have a great potential in the treatment of various MDR cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Genome-wide scan of reading ability in affected sibling pairs with attention-deficit/hyperactivity disorder: unique and shared genetic effects.

Attention-deficit/hyperactivity disorder (ADHD) and reading disability (RD) are common highly heritable disorders of childhood, which frequently co-occur. Data from twin and family studies suggest that this overlap is, in part, due to shared genetic underpinnings. Here, we report the first genome-wide linkage analysis of measures of reading ability in children with ADHD, using a sample of 233 affected sibling pairs who previously participated in a genome-wide scan for susceptibility loci in ADHD. Quantitative trait locus (QTL) analysis of a composite reading factor defined from three highly correlated reading measures identified suggestive linkage (multipoint maximum lod score, MLS>2.2) in four chromosomal regions. Two regions (16p, 17q) overlap those implicated by our previous genome-wide scan for ADHD in the same sample: one region (2p) provides replication for an RD susceptibility locus, and one region (10q) falls approximately 35 cM from a modestly highlighted region in an independent genome-wide scan of siblings with ADHD. Investigation of an individual reading measure of Reading Recognition supported linkage to putative RD susceptibility regions on chromosome 8p (MLS=2.4) and 15q (MLS=1.38). Thus, the data support the existence of genetic factors that have pleiotropic effects on ADHD and reading ability--as suggested by shared linkages on 16p, 17q and possibly 10q--but also those that appear to be unique to reading--as indicated by linkages on 2p, 8p and 15q that coincide with those previously found in studies of RD. Our study also suggests that reading measures may represent useful phenotypes in ADHD research. The eventual identification of genes underlying these unique and shared linkages may increase our understanding of ADHD, RD and the relationship between the two.

Adolescent↗

Transmission disequilibrium testing of dopamine-related candidate gene polymorphisms in ADHD: confirmation of association of ADHD with DRD4 and DRD5.

Attention-deficit hyperactivity disorder (ADHD) is one of the most common childhood behavioral disorders. Genetic factors contribute to the underlying liability to develop ADHD. Reports implicate variants of genes important for the synthesis, uptake, transport and receptor binding of dopamine in the etiology of ADHD, including DRD4, DAT1, DRD2, and DRD5. In the present study, we genotyped a large multiplex sample of ADHD affected children and their parents for polymorphisms in genes previously reported to be associated with ADHD. Associations were tested by the transmission disequilibrium test (TDT). The sample is sufficient to detect genotype relative risks (GRRs) for putative risk alleles. The DRD4 gene 120-bp insertion/deletion promoter polymorphism displayed a significant bias in transmission of the insertion (chi(2)=7.58, P=0.006) as suggested by an analysis of a subset of these families. The seven repeat allele of the DRD4 48-bp repeat polymorphism (DRD4.7) was not significantly associated with ADHD in the large sample in contrast to our earlier findings in a smaller subset. We replicate an association of a dinucleotide repeat polymorphism near the DRD5 gene with ADHD by showing biased nontransmission of the 146-bp allele (P=0.02) and a trend toward excess transmission of the 148-bp allele (P=0.053). No evidence for an association was found for polymorphisms in DRD2 or DAT1 in this sample. The DRD5 146-bp (DRD5.146) allele and the DRD4 240-bp (DRD4.240) allele of the promoter polymorphism emerge as the two DNA variants showing a significant association in this large sample of predominantly multiplex families with ADHD, with estimated GRRs of 1.7 and 1.37, respectively.

Adolescent↗

Expression of CD59 on lymphocyte and the subsets and its potential clinical application for paroxysmal nocturnal hemoglobinuria diagnosis.

Lymphocyte analysis has not been accepted as an indicator for the clinical diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), because of diverse CD59 expression, even in the healthy individuals. We studied the expression of CD59 on lymphocytes and lymphocyte subsets in healthy individuals and PNH patients by flow cytometry. In 50 healthy individuals, granulocytes generally showed a single-cell population that was strongly positive for CD59. Lymphocytes showed, in addition to the population of positive cells, a small population of weakly positive cells, while CD3+ T-cells showed a large population of weakly positive cells. Natural killer (NK)-cells showed a profile similar to CD3+ T-cells. CD4+ T-cells showed a larger population of strongly positive cells, as compared with CD3+ T-cells, while CD8+ T-cells showed a smaller population of strongly positive cells. CD20+ B-cells showed a similar profile to granulocytes. CD59 expression studies in all our PNH patients showed that B-cells expressed CD59 at levels similar to granulocytes, which were lower than those of T-cells and NK-cells. Therefore, the lymphocyte population with low level CD59 expression in healthy controls corresponds to T-cells (especially CD8+ T-cells) and NK-cells. CD59 expression on B-cells could thus be used as a new marker for the diagnosis of PNH.

Antigens, CD↗

EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways.

Epidermal growth factor (EGF) is present in the maternal-fetal environment and has an important role in placental development. Matrix metalloproteinase-9 (MMP-9) expression/activation is a pre-requisite in extravillous trophoblast invasion. Whereas EGF up-regulates MMP-9 activity in a variety of cell types, there is no direct evidence for the stimulation of MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1) secretion by EGF in extravillous trophoblasts. In addition, the signalling pathways involved in this regulation are not clear. In the present study, we have examined the possible involvement of the phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways in the regulation of the MMP-9/TIMP-1 system by EGF in vitro. We used a well-established invasive extravillous trophoblast cell line (HTR8/Svneo) and measured gene and protein expression by semi-quantitative RT-PCR and western analysis respectively. MMP activity was determined by zymography. We showed for the first time that EGF activated both PI3K/Akt and MAPK/extracellular-signal regulated kinase (ERK) signalling in HTR8/SVneo, and increased both MMP-9 and TIMP-1 mRNAs and protein concentrations. Interfering with either signalling pathway via PI3K inhibitor LY294002 or MEK inhibitor U0126 in EGF-stimulated HTR8/SVneo cells blocked the induction of MMP-9 and TIMP-1. LY294002 inhibited Akt phosphorylation, but had no effect on ERK phosphorylation; U0126 suppressed ERK phosphorylation without interfering with the phosphorylation of Akt. In addition, expression of constitutively active Akt (Myr-Akt1, Myr-Akt2, Myr-Akt3) was not sufficient to induce proMMP-9 and TIMP-1 secretion. Our results suggest that the activation of both PI3K and MAPK pathways in extravillous trophoblasts is necessary for the up-regulation of MMP-9 and TIMP-1 expression by EGF.

Blotting, Western↗

Treatment of oilfield wastewater by Fenton's process.

A combination of coagulation and Fenton's process was used for the removal of total oxygen carbon (TOC) from oilfield wastewater. Compared with aluminium sulfate, ferric coagulant had better TOC removal efficiency at the same mass dosage. In Fenton's process, the effect of H2O2 and Fe2+ dose on the removal of TOC was studied. The optimum conditions required for TOC removal were an Fe3+ concentration of 40-50 mg/L, an H2O2 dose of 50 mmol/L and an Fe2+ concentration of 1.0 mmol/L. GC-MS chromatographic analysis indicated that most of the alkyl hydrocarbons of carbon numbers < 21 were removed in the first minute of Fenton's process mainly through adsorption. Alkyl hydrocarbons and phenols were oxidized almost completely following 120 min of treatment. The pathway of newly formed intermediates in Fenton's process was proposed on the basis of the GC/MS chromatogram.

Hydrocarbons↗

An EST survey of the sugarcane transcriptome.

Its large genome and high polyploidy makes sugarcane (Saccharum spp.) a singularly challenging crop to study and improve using genetic approaches. To provide large numbers of functionally characterized candidate genes that might be tested for direct association (rather than distant linkage) with economically important traits, we sequenced the 5' ends of 9,216 clones from three cDNA libraries (apex, leaf and mature internode), representing 3,401 non-redundant sequences. About 57% of these sequences could be assigned a tentative function based on statistically significant similarity to previously characterized proteins or DNA sequences. Another 28% corresponded to previously identified, but uncharacterized, sequences. Some of the remaining unidentified sequences were predicted to be genes which could potentially be new to plants or unique to sugarcane. Comparisons of the sugarcane ESTs to a large sorghum EST database revealed similar compositions of expressed genes between some different tissues. Comparison to a detailed Arabidopsis protein database showed some highly conserved sequences, which might be useful DNA markers for pan-angiosperm comparative mapping. These EST sequences provide a foundation for many new studies to accelerate isolation of agronomically important genes from the cumbersome sugarcane genome.

Base Sequence↗

Cross section for the H+H2O abstraction reaction: experiment and theory.

The absolute value of the cross section for the abstraction reaction between fast H atoms and H2O has been determined experimentally at a mean collision energy of 2.46 eV. The OH population distribution at the same mean energy has also been determined. The new measurements are compared with state-of-the-art quantum mechanical and quasiclassical scattering calculations on the most recently developed potential energy surface.

Journal Article↗

The presence of insertion elements IS861 and IS1548 in group B streptococci.

The presence of insertion elements (IS) IS861 and IS1548 in the collection of 211 Streptococcus agalactiae strains isolated from pregnant women and dairy cows was assayed. IS861 was found in 67 human strains (59%) and 36 bovine strains (37%), IS1548 in 13 human strains (12%) and 16 bovine strains (16%). Two combinations, IS861+ IS1548- and IS861- IS1548-, were widely distributed in both human and bovine strains. The copy number and the restriction fragment length polymorphism (RFLP) of the two IS were determined in human group B streptococcus (GBS) strains. A minimum of 8 copies of IS1548 were detected in GBS strains while the copy number of IS861 varied from 1 to 9. The number of different hybridizing patterns with IS861 and IS1548 probes was 9 and 6, respectively. These hybridization patterns were divided into several clusters. All strains with IS were also clustered according to pulsed field-gel electrophoresis (PFGE) patterns. A correlation was found between the results of PFGE- and IS-based clustering.

Animals↗

Biosorption of reactive dyes by the mycelium pellets of a new isolate of Penicillium oxalicum.

Three reactive dyes were rapidly adsorbed by the mycelium pellets of Penicillium oxalicum. Dye removal of Reactive Blue 19 was up to 60% in 10 min and 91% in 80 min. Dye adsorption isotherms fitted Langmuir model well and the maximum adsorption capacities at 20 degrees C were calculated to be 160 mg g(-1) for Reactive Blue 19, 122 mg g(-1) for Reactive Red 241 and 137 mg g(-1) for Reactive Yellow 145, respectively. The pellets exhibited a high dye adsorption capacity (80-180 mg g(-1)) for all of the 3 dyes over a wide pH range (pH 2-10), and the maximum adsorption was obtained at pH 2. The adsorption capacity was mildly increased by increasing salinity.

Adsorption↗

Dissociation of T helper type 2 cytokine-dependent airway lesions from signal transducer and activator of transcription 6 signalling in experimental chronic asthma.

BACKGROUND: Type 2 T helper lymphocytes (Th2 cells) and their cytokine products are important in the pathogenesis of asthma. OBJECTIVE: To examine the contribution of the signal transducer and activator of transcription (STAT) 6 pathway, involved in Th2 cytokine signalling, to the development of lesions of chronic asthma. METHODS: BALB/c mice sensitized to ovalbumin were chronically challenged by inhalational of low mass concentrations of antigen for 6 weeks. Airway lesions in wild-type mice were compared with those in STAT6-deficient mice and in IL-4/13 double-deficient mice by histomorphometry and immunohistochemistry. Airway responses to methacholine were evaluated by whole-body plethysmography. Cytokine production by peribronchial lymph node cells was quantified by enzyme immunoassay. RESULTS: STAT6-/- mice developed a variety of airway lesions that were at least equivalent to those in wild-type mice, including accumulation of intraepithelial eosinophils and of chronic inflammatory cells in the lamina propria, subepithelial fibrosis and epithelial thickening. In addition, STAT6-/- mice exhibited exaggerated airway hyper-reactivity (AHR) compared to wild-type animals. This was despite a shift from a Th2 to a Th1 pattern of immunoglobulin production by plasma cells in the inflammatory infiltrate and diminished mucous cell hyperplasia/metaplasia, together with increased production of IFN-gamma by peribronchial lymph node cells, consistent with absence of signalling via the STAT6 pathway. In contrast, gene-targeted IL-4/13-/- mice exhibited markedly diminished eosinophil recruitment and airway remodelling, as well as absence of AHR. CONCLUSIONS: In this model, the effects of STAT6 deficiency were in marked contrast to the suppression of inflammation and AHR described in models of allergic bronchopulmonary inflammation. These results, which provide evidence of STAT6-independent AHR in an inhalational challenge model of chronic asthma, emphasize the critical effector roles of IL-4 and IL-13, as well as the need to use appropriate models to understand cytokine signalling pathways that may be potential therapeutic targets in asthma.

Animals↗

Using ammonium-tolerant yeast isolates: Candida halophila and Rhodotorula glutinis to treat high strength fermentative wastewater.

Two ammonium-tolerant yeast strains were isolated from sludge samples contaminated with monosodium glutamate manufacturing wastewater and were identified as Candida haplophila and Rhodotorula glutinis. The tolerance of the two yeast isolates to ammonia and their chemical oxygen demand (COD) removal perfomances were evaluated under batch and bench-scale conditions. The mixture of the two isolates was found to grow well in an artificial medium containing 25% (NH4)2SO4 and could effectively remove COD from monosodium glutamate wastewater even when the concentrations of NH4+-N and free NH3-N reached as high as 18,977 and 879 mg l(-1) respectively. A fixed-bed yeast reactor, which was initially inoculated with the yeast mixture, permitted a constant COD removal rate of over 80% during a period of near 2-month continuous running even when the influent COD was increased from 8,000 to 25,000 mg l(-1). The effluent was accompanied with suspended solids (SS) of over 4,500 mg l(-1), which was mainly composed of yeast cells and could be considered as a source of animal forage additive. The residual COD of effluents from the yeast reactor could be further reduced to under 500 mg l(-1) by a combination process of activated sludge treatment and coagulation technologies.

Adaptation, Physiological↗

Effects of green tea on carcinogen-induced hepatic CYP1As in C57BL/6 mice.

Green tea (GT) drinking showed chemopreventive effects on various cancers. In addition, inhibition of CYP1A activity by green tea components--polyphenols--has been suggested as a chemoprevention against carcinogens that were bioactivated by CYP1As. Therefore, any changes in hepatic CYP1As may be considered as a biomarker for GT chemoprevention and clarify whether whole GT is chemopreventive for the population who are exposed to CYP1A specifically-bioactivated carcinogens. In this study, we investigated the changes in CYP1A levels by pre- and concurrent GT drinking against a CYP1A-inducing carcinogen, 3-methylcholanthrene (MC), in aryl hydrocarbon receptor responsive C57 BL/6 mice. We found that GT drinking itself induced hepatic CYP1As and enhanced MC-induced ethoxyresorufin-O-demethylase (EROD) activity (P<0.05). However, our studies of CYP1A monoclonal antibody and western blots revealed that the enhanced hepatic EROD activity by GT did not come from CYP1As. Therefore, our results suggest that GT may work to biotransform CYP1A inducing carcinogens into non-carcinogenic metabolites by modulation of other microsomal enzymes rather than CYP1As. In addition, the mechanism of GT chemoprevention may be different from that of GT components, such as polyphenols that reduce CYP1As activity.

Administration, Oral↗

The central role of human chorionic gonadotropin in the formation of human placental syncytium.

A number of trophoblast products, including human chorionic gonadotropin (hCG), can increase the formation of human placental syncytium through the differentiation of mononuclear cytotrophoblasts. The present study investigated the central role of hCG in this process by using antisense receptor phosphorothioate oligodeoxynucleotides (ODNs). Culturing cytotrophoblasts with the hCG/LH receptor antisense, but not sense, ODN resulted in a significant decrease in receptor protein levels and inhibited spontaneous as well as exogenous hCG induced increase in differentiation. The hCG/LH receptor antisense ODN also inhibited epidermal growth factor (EGF), TGF-alpha, leukemia inhibitory factor (LIF), but not 8-bromo-cAMP, induced increases in differentiation, suggesting that hCG is required for EGF, TGF-alpha and LIF, but not for the cAMP actions. Although antisense EGF receptor and LIF receptor ODNs inhibited EGF and LIF induced increase in differentiation, respectively, they were ineffective against hCG, suggesting that they use separate pathways, but they both converge on a common pathway requiring the hCG actions. Mechanism of action studies revealed that EGF treatment activates its receptors and MAPK, both of which are required for EGF to increase the differentiation, cAMP levels and activate protein kinase A. In summary, our results demonstrate that hCG is an autocrine and paracrine regulator that is required for EGF, TGF-alpha, and LIF, but not for cAMP to increase human placental syncytium formation. Direct activation of protein kinase A seems to bypass the hCG pathway, perhaps by targeting genes associated with the differentiation.

8-Bromo Cyclic Adenosine Monophosphate↗

Comparison of capillary electrophoresis and reversed-phase liquid chromatography for determination of the enantiomeric purity of an M3 antagonist.

The chiral separation of an M3 antagonist was investigated using capillary electrophoresis (CE) with various sulfated cyclodextrins and by reversed-phase liquid chromatography with derivatized cellulose, derivatized amylose, and two protein stationary phases. Operational parameters for each technique, such as the concentration of the chiral selectors, background electrolyte (or mobile phase) pH and type, organic modifiers, injection mode and temperature were varied in order to achieve a desired elution order and to meet a 0.1% limit of quantitation (LOQ) criteria. Based on the advantages and disadvantages of each technique, a practical CE method using sulfated gamma-cyclodextrin was selected. The method was validated in terms of linearity, LOQ, accuracy, ruggedness and precision.

Buffers↗

Erythropoietin production from CHO cells grown by continuous culture in a fluidized-bed bioreactor.

A Chinese hamster ovary (CHO) cell line that expresses human erythropoietin (huEPO) was in a 2-L Cytopilot fluidized-bed bioreactor with 400 mL macroporous Cytoline-1 microcarriers and a variable perfusion rate of serum-free and protein-free medium for 48 days. The cell density increased to a maximum of 23 x 10(6) cells/mL, beads on day 27. The EPO concentration increased to 600 U/mL during the early part of the culture period (on day 24) and increased further to 980 U/mL following the addition of a higher concentration of glucose and the addition of sodium butyrate. The EPO concentration was significantly higher (at least 2x than that in a controlled stirred-tank bioreactor, in a spinner flask, or in a stationary T-flask culture. The EPO accumulated to a total production of 28,000 kUnits over the whole culture period. The molecular characteristics of EPO with respect to size and pattern of glycosylation did not change with scale up. The pattern of utilization and production of 18 amino acids was similar in the Cytopilot culture to that in a stationary batch culture in a T-flask. The concentration of ammonia was maintained at a low level (< 2 mM) over the entire culture period. The specific rate of consumption of glucose, as well as the specific rates of production of lactate and ammonia, were constant throughout the culture period indicating a consistent metabolic behavior of the cells in the bioreactor. These results indicate the potential of the Cytopilot bioreactor culture system for the continuous production of a recombinant protein over several weeks.

Amino Acids↗