Showing papers by "Southwest University published in 2007"

Microarray-based gene expression profiles in multiple tissues of the domesticated silkworm, Bombyx mori

Abstract: We designed and constructed a genome-wide microarray with 22,987 70-mer oligonucleotides covering the presently known and predicted genes in the silkworm genome, and surveyed the gene expression in multiple silkworm tissues on day 3 of the fifth instar. Clusters of tissue-prevalent and tissue-specific genes and genes that are differentially expressed in different tissues were identified, and they reflect well major tissue-specific functions on the molecular level. The data presented in this study provide a new resource for annotating the silkworm genome.

Increased insect virulence in Beauveria bassiana strains overexpressing an engineered chitinase.

Abstract: Entomopathogenic fungi are currently being used for the control of several insect pests as alternatives or supplements to chemical insecticides. Improvements in virulence and speed of kill can be achieved by understanding the mechanisms of fungal pathogenesis and genetically modifying targeted genes, thus improving the commercial efficacy of these biocontrol agents. Entomopathogenic fungi, such as Beauveria bassiana, penetrate the insect cuticle utilizing a plethora of hydrolytic enzymes, including chitinases, which are important virulence factors. Two chitinases (Bbchit1 and Bbchit2) have previously been characterized in B. bassiana, neither of which possesses chitin-binding domains. Here we report the construction and characterization of several B. bassiana hybrid chitinases where the chitinase Bbchit1 was fused to chitin-binding domains derived from plant, bacterial, or insect sources. A hybrid chitinase containing the chitin-binding domain (BmChBD) from the silkworm Bombyx mori chitinase fused to Bbchit1 showed the greatest ability to bind to chitin compared to other hybrid chitinases. This hybrid chitinase gene (Bbchit1-BmChBD) was then placed under the control of a fungal constitutive promoter (gpd-Bbchit1-BmChBD) and transformed into B. bassiana. Insect bioassays showed a 23% reduction in time to death in the transformant compared to the wild-type fungus. This transformant also showed greater virulence than another construct (gpd-Bbchit1) with the same constitutive promoter but lacking the chitin-binding domain. We utilized a strategy where genetic components of the host insect can be incorporated into the fungal pathogen in order to increase host cuticle penetration ability.

‘GM‐gene‐deletor’: fused loxP‐FRT recognition sequences dramatically improve the efficiency of FLP or CRE recombinase on transgene excision from pollen and seed of tobacco plants

Abstract: Pollen- and seed-mediated transgene flow is a concern in plant biotechnology. We report here a highly efficient 'genetically modified (GM)-gene-deletor' system to remove all functional transgenes from pollen, seed or both. With the three pollen- and/or seed-specific gene promoters tested, the phage CRE/loxP or yeast FLP/FRT system alone was inefficient in excising transgenes from tobacco pollen and/or seed, with no transgenic event having 100% efficiency. When loxP-FRT fusion sequences were used as recognition sites, simultaneous expression of both FLP and CRE reduced the average excision efficiency, but the expression of FLP or CRE alone increased the average excision efficiency, with many transgenic events being 100% efficient based on more than 25,000 T(1) progeny examined per event. The 'GM-gene-deletor' reported here may be used to produce 'non-transgenic' pollen and/or seed from transgenic plants and to provide a bioconfinement tool for transgenic crops and perennials, with special applicability towards vegetatively propagated plants and trees.

Determinants of Water Conservation Intention in Blagoevgrad, Bulgaria

Abstract: Blagoevgrad, Bulgaria, has faced frequent water shortages during the past decade, but little effort has been made to encourage or implement water conservation among household water users. We employ the theory of planned behavior (TBP) as a theoretical framework in evaluating determinates of intention to implement a set of specific water conservation behaviors among Blagoevgrad residents. In addition to the TPB variables, we consider the effects of sociodemographics, environmental attitudes, information possession, and concern over future shortages on water conservation intention. The analysis is based on 728 questionnaires collected during the summer of 2003. The TPB variables all showed positive and significant correlation with water conservation intention. Self-perceived knowledge of climate change was also significantly related to intention. Environmental attitudes and concern over future shortages were significant but relatively weak determinants.

Testing the viability of the interacting holographic dark energy model by using combined observational constraints

Abstract: Using the data coming from the new 182 gold type Ia supernova samples, the shift parameter of the cosmic microwave background given by the three-year Wilkinson Microwave Anisotropy Probe observations, and the baryon acoustic oscillation measurement from the Sloan digital sky survey, H(z) and lookback time measurements, we have performed a statistical joint analysis of the interacting holographic dark energy model. Consistent parameter estimations show us that the interacting holographic dark energy model is a viable candidate for explaining the observed acceleration of our universe.

Avian-human influenza epidemic model.

Abstract: A mathematical model is proposed to interpret the spread of avian influenza from the bird world to the human world. Our mathematical model warns that two types of the outbreak of avian influenza may occur if the humans do not prevent the spread of avian influenza. Moreover, it suggests that we cannot feel relieved although the total infected humans are kept at low level. In order to prevent spread of avian influenza in the human world, we must take the measures not only for the birds infected with avian influenza to exterminate but also for the humans infected with mutant avian influenza to quarantine when mutant avian influenza has already occurred. In particular, the latter measure is shown to be important to stop the second pandemic of avian influenza.

GhDET2, a steroid 5alpha-reductase, plays an important role in cotton fiber cell initiation and elongation.

Abstract: Cotton (Gossypium hirsutum L.) fibers, one of the most important natural raw materials for textile industry, are highly elongated trichomes from epidermal cells of cotton ovules. DET2, an Arabidopsis steroid 5d-reductase, is considered to catalyze a major rate-limiting in brassinosteroid (BR) biosynthesis. To understand the role of BRs in cotton fiber development, GhDET2, which putatively encodes a steroid 5alpha-reductase by sequence comparison, was cloned from developing fiber cells. In vitro assessment of GhDET2 protein activity confirmed that GhDET2 encodes a functional steroid 5alpha-redutase. High levels of GhDET2 transcript were detected during the fiber initiation stage and the fiber rapid elongation stage. Antisense-mediated suppression of GhDET2 inhibited both fiber initiation and fiber elongation. Similarly, treating cultured ovules with finasteride, a steroid 5alpha-reductase inhibitor, reduced fiber elongation. Inhibition of fiber cell elongation by expression of antisense GhDET2 or the finasteride treatment could be reversed by epibrassinolide, a biologically active BR. Furthermore, seed coat-specific expression of GhDET2 increased fiber number and length. Therefore, GhDET2 and BRs play a crucial role in the initiation and elongation of cotton fiber cells, suggesting that modulation of BR biosynthesis factors may improve fiber quality or yield.

Propagation of HBV with spatial dependence.

Abstract: A mathematical model is proposed to simulate the hepatitis B virus (HBV) infection with spatial dependence. The existence of traveling waves is established via the geometric singular perturbation method. Numerical simulations show that the model admits non-monotone traveling profiles. Influences of various parameters on the minimum wave speed are also discussed.

Testing the viability of the interacting holographic dark energy model by using combined observational constraints

Abstract: Using the data coming from the new 182 Gold type Ia supernova samples, the shift parameter of the Cosmic Microwave Background given by the three-year Wilkinson Microwave Anisotropy Probe observations, and the baryon acoustic oscillation measurement from the Sloan Digital Sky Survey, $H(z)$ and lookback time measurements, we have performed a statistical joint analysis of the interacting holographic dark energy model. Consistent parameter estimations show us that the interacting holographic dark energy model is a viable candidate to explain the observed acceleration of our universe.

Magnetic Control of an Electrochemical Microfluidic Device with an Arrayed Immunosensor for Simultaneous Multiple Immunoassays

Abstract: Background: Methods based on magnetic bead probes have been developed for immunoassay, but most involve complicated labeling or stripping procedures and are unsuitable for routine use. Methods: We synthesized magnet core/shell NiFe2O4/SiO2 nanoparticles and fabricated an electrochemical magnetic controlled microfluidic device for the detection of 4 tumor markers. The immunoassay system consisted of 5 working electrodes and an Ag/AgCl reference electrode integrated on a glass substrate. Each working electrode contained a different antibody immobilized on the NiFe2O4/SiO2 nanoparticle surface and was capable of measuring a specific tumor marker using noncompetitive electrochemical immunoassay. Results: Under optimal conditions, the multiplex immunoassay enabled the simultaneous detection of 4 tumor markers. The sensor detection limit was 5 μg/L (or >5 kunits/L), respectively. No nonspecific adsorption was observed during a series of procedures to detect target proteins, and electrochemical cross-talk (CV) between neighboring sites was <10%. Conclusion: This immunoassay system offers promise for label-free, rapid, simple, cost-effective analysis of biological samples. Importantly, the chip-based immunosensor could be suitable for use in the mass production of miniaturized lab-on-a-chip devices and open new opportunities for protein diagnostics and biosecurity.

A Novel Type of P450c17 Lacking the Lyase Activity Is Responsible for C21-Steroid Biosynthesis in the Fish Ovary and Head Kidney

Abstract: Cytochrome P450c17 is the single enzyme that mediates the 17 alpha-hydroxylase and 17, 20 lyase activities during the biosynthesis of steroid hormones in the gonads and adrenal gland. However, the mechanism underlying its dual action continues to be a controversy in the field of steroidogenesis in fish. In an attempt to resolve this issue, we identified a novel type of P450c17 (P450c17-II) by an in silico analysis from the genomes of six fish species. We cloned P450c17-II from tilapia and medaka, and comparison with the conventional P450c17-I revealed that they differ in gene structure and enzymatic activity. Enzymatic assays by thin-layer chromatography revealed that P450c17-II possesses only the 17 alpha-hydroxylase activity without any 17, 20 lyase activity, unlike P450c17-I, which has both these activities. In testis, both P450c17-I and -II express in the interstitial cells. Remarkable differences, revealed by in situ hybridization, in the expression patterns of the P450c17-I and -II in the ovary and head kidney of tilapia during various stages of development strongly suggest that P450c17-I is responsible for the synthesis of estradiol-17beta in the ovary, whereas P450c17-II is required for the production of C21 steroids such as cortisol in the head kidney. More interestingly, a temporally controlled switching is observable in the expression of these two genes during the steroidogenic shift from estradiol-17beta to the C21 steroid, 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (maturation-inducing hormone of fish oocytes) in the fish ovary, revealing a role for P450c17-II in the production of hormones that induce oocyte maturation in fish.

Magnetic-Core/Porous-Shell CoFe2O4/SiO2 Composite Nanoparticles as Immobilized Affinity Supports for Clinical Immunoassays†

Abstract: This study demonstrates a novel approach towards the development of advanced protein assay systems based on physically functionalized, magnetic-core/porous-shell CoFe 2 O 4 /SiO 2 composite nanoparticles. The preparation, characterization, and measurement of the relevant properties of the protein assay system is discussed, and the system is used for the detection of cancer antigen 15-3 (CA 15-3, used as a model here) in clinical immunoassays. The protein assay system, based on nanometersized magnetic cores and silica shells, shows good adsorption properties for the selective attachment of CA 15-3 antibodies specific to CA 15-3. The core/shell nanostructures exhibit good magnetic properties, which enables their integration into a quartz crystal microbalance (QCM) detection cell with the help of a permanent magnet. Under optimal conditions, the resulting immunoassay system presents a good QCM response for the detection of CA 15-3, and allows the detection of CA 15-3 at concentrations as low as 1.5 U mL -1 (U: units). Importantly, the proposed protein assay system can be extended to the detection of other antigens and biological compounds.

The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters.

Abstract: Here we report the effect of the 35S promoter sequence on activities of the tissue- and organ-specific gene promoters in tobacco plants. In the absence of the 35S promoter sequence the AAP2 promoter is active only in vascular tissues as indicated by expression of the AAP2:GUS gene. With the 35S promoter sequence in the same T-plasmid, transgenic plants exhibit twofold to fivefold increase in AAP2 promoter activity and the promoter becomes active in all tissue types. Transgenic plants hosting the ovary-specific AGL5:iaaM gene (iaaM coding an auxin biosynthetic gene) showed a wild-type phenotype except production of seedless fruits, whereas plants hosting the AGL5:iaaM gene along with the 35S promoter sequence showed drastic morphological alterations. RT-PCR analysis confirms that the phenotype was caused by activation of the AGL5:iaaM gene in non-ovary organs including roots, stems and flowers. When the pollen-, ovule- and early embryo-specific PAB5:barnase gene (barnase coding a RNase gene) was transformed, the presence of 35S promoter sequence drastically reduced transformation efficiencies. However, the transformation efficiencies were restored in the absence of 35S promoter, indicating that the 35S promoter might activate the expression of PAB5:barnase in non-reproductive organs such as calli and shoot primordia. Furthermore, if the 35S promoter sequence was replaced with the NOS promoter sequence, no alteration in AAP2, AGL5 or PAB5 promoter activities was observed. Our results demonstrate that the 35S promoter sequence can convert an adjacent tissue- and organ-specific gene promoter into a globally active promoter.

Combination of features from skin pattern and ABCD analysis for lesion classification.

Abstract: Background/Purpose: It is known that the standard features for lesion classification are ABCD features, that is, asymmetry, border irregularity, colour variegation and diameter of lesion. However, the observation that skin patterning tends to be disrupted by malignant but not by benign skin lesions suggests that measurements of skin pattern disruption on simply captured white light optical skin images could be a useful contribution to a diagnostic feature set. Previous work using both skin line direction and intensity for lesion classification was encouraging. But these features have not been combined with the ABCD features. This paper explores the possibility of combing features from skin pattern and ABCD analysis to enhance classification performance. Methods: The skin line direction and intensity were extracted from a local tensor matrix of skin pattern. Meanwhile, ABCD analysis was conducted to generate six features. They were asymmetry, border irregularity, colour (red, green and blue) variegations and diameter of lesion. The eight features of each case were combined using a principal component analysis (PCA) to produce two dominant features for lesion classification. Results: A larger set of images containing malignant melanoma (MM) and benign naevi were processed as above and the scatter plot in a two-dimensional dominant feature space showed excellent separation of benign and malignant lesions. An ROC (receiver operating characteristic) plot enclosed an area of 0.94. Conclusions: The classification results showed that the individual features have a limited discrimination capability and the combined features were promising to distinguish MM from benign lesion.

Molecular cloning of doublesex and mab-3-related transcription factor 1, forkhead transcription factor gene 2, and two types of cytochrome P450 aromatase in Southern catfish and their possible roles in sex differentiation.

Abstract: To address the roles of doublesex and mab-3-related transcription factor 1 (Dmrt1), forkhead transcription factor gene 2 (Foxl2), and aromatase in sex differentiation of Southern catfish, the cDNA sequences of these genes were isolated from the gonads. Dmrt1a and Dmrt1b were found to be expressed in the gonads, being higher in the testis. A low expression level of Dmrt1b was also detected in the intestine and kidney of the male. Foxl2 was found to be expressed extensively in the brain (B), pituitary (P), gill and gonads (G), with the highest level in the ovary, indicating the possible involvement of Foxl2 in the B-P-G axis. Cytochrome P450 (Cyp)19b was found to be expressed in the brain, spleen, and gonads, while Cyp19a was only expressed in the gonads and spleen. All-female Southern catfish fry were treated with fadrozole (F), tamoxifen (TAM), and 17beta-estradiol (E2) respectively, from 5 to 25 days after hatching (dah). The expression levels of these genes were measured at 65 dah. In the F-, TAM-, and FTAM-treated groups, Dmrt1a and Dmrt1b were up-regulated in the gonad, whereas Foxl2 and Cyp19a were down-regulated, while the expression of Cyp19b in the gonad remained unchanged. Furthermore, down-regulation of Foxl2 and Cyp19b was also detected in the brain. In the E2-treated group, Dmrt1a and Dmrt1b were down-regulated to an undetectable level in the gonad, whereas Foxl2 and Cyp19b were up-regulated in the brain. Consistent with the observed changes in the expressions of these genes, 56, 70, and 80% sex-reversed male individuals were obtained in the F-, TAM-, and F + TAM-treated groups respectively. These results indicate the significant roles of Dmrt1, Foxl2, and Cyp19 in the sex differentiation of Southern catfish.

Effect of organic matter and pH on mercury release from soils

Abstract: An investigation was conducted on the effect of organic matter (OM) and pH on mercury (Hg) release from soils. Hg release flux was measured using the dynamic flux chamber (DFC) combined with the Lumex® multifunctional mercury analyzer in both laboratory experiment and field monitoring. The results showed that Hg emission from the OM-added soils was apparently low because of the high affinity of OM to Hg, resulting in the reverse order as the amount of OM addition. Meanwhile, Hg release flux from different pH value soils exhibited the same trend for both Hg 2+ and Hg 2 2+ treatment, increasing the Hg flux with pH value of soils increasing. The trend of Hg release in the pH dependence experiment has been well in agreement with that from the field test. In addition, Hg release seemed to be related to its species in the soil, the flux from Hg 2+ -added soil was obviously higher than that of Hg 2 2+ -added soil by the laboratory experiment.

A regulator of a G protein signalling (RGS) gene, cag8, from the insect-pathogenic fungus Metarhizium anisopliae is involved in conidiation, virulence and hydrophobin synthesis.

Abstract: Regulators of the G protein signalling (RGS) pathway have been implicated in the control of a diverse array of cellular functions, including conidiation in filamentous fungi. However, the regulatory processes involved in conidiation in insect-pathogenic fungi are poorly understood. Since conidia are the infective propagules in these fungi, an understanding of the regulatory processes involved in conidiation is essential to the development of an effective biocontrol fungus. Here, the cloning and characterization of an RGS protein gene, cag8 (conidiation-associated gene), from the insect-pathogenic fungus Metarhizium anisopliae is reported. Phylogenetic analysis showed that CAG8 was orthologous to the RGS protein FlbA from Aspergillus nidulans. Complementation of A. nidulans ΔflbA, which cannot conidiate, with M. anisopliae cag8 restored conidiation. Gene disruption of cag8 in M. anisopliae resulted in the lack of conidia on agar plates and on infected insects, reduced mycelial growth, decreased virulence, lysis during growth in liquid medium as well as lack of pigmentation and irregularly shaped blastospores. Transcript levels of ssgA (hydrophobin-encoding gene) were markedly reduced in a Δcag8 strain, while pr1A (subtilisin-like protease) transcription was unaffected. These results suggest that cag8 is involved in the modulation of conidiation, virulence and hydrophobin synthesis in M. anisopliae.