In JoVE (1)

Other Publications (166)

Articles by Kongming Wu in JoVE

 JoVE Immunology and Infection

Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm

1State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 2Tobacco Research Institute, Chinese Academy of Agricultural Sciences, 3Crop and Environment Sciences, Harper Adams University

JoVE 55534

Other articles by Kongming Wu on PubMed

Resistance Monitoring of Helicoverpa Armigera (Lepidoptera Noctuidae) to Bacillus Thuringiensis Insecticidal Protein in China

Journal of Economic Entomology. Aug, 2002  |  Pubmed ID: 12216827

Sensitivities of Helicoverpa armigera (Hübner) field populations to Bacillus thuringiensis (Bt) insecticidal protein CrylAc were monitored during 1998-2000 in China. A total of 41 strains was sampled, and most of them were collected from Bt cotton planting regions. The range of IC50 values (concentration producing 50% inhibition of larval development to third instar) among different populations in 1998,1999, and 2000 were 0.020-0.105 microg/ml, 0.016-0.099 microg/ml, and 0.016-0.080 microg/ml, respectively. Diagnostic concentration studies (IC99) showed that the percentage of individuals reaching third instar ranged from 0 to 4.35%, with only eight of the 41 tested populations showing values above 0%. Also interesting was a trend over successive years in which fewer populations contained individuals that survived the diagnostic concentration by reaching third instar. Considering these data, it was determined that the field populations sampled during the 3-yr study were susceptible to CrylAc protein, and that movement toward resistance among H. armigera populations was not apparent.

Evaluation of the Natural Refuge Function for Helicoverpa Arnigera (Lepidoptera: Noctuidae) Within Bacillus Thuringiensis Transgenic Cotton Growing Areas in North China

Journal of Economic Entomology. Aug, 2002  |  Pubmed ID: 12216828

The density of Helicoverpa armigera (Hübner) populations on Bacillus thuringiensis Berliner (Bt) transgenic cotton, corn, peanut, and soybean; differences in its development on Bt cotton and common (nontransgenic) cotton; and the potential for mating among populations from Bt cotton fields and other crop fields were investigated in the suburbs of Xinxiang City (Henan Province) and Langfang City (Hebei Province) in the southern and northern parts of north China, respectively. Although development of H. armigera on Bt cotton was much slower than on common cotton, there was a still high probability of mating between populations from Bt cotton and other sources due to the scattered emergence pattern of H. armigera adults, and overlap of the second and third generations. In a cotton and corn growing region, early and late planted corn provided suitable refugia for the third and fourth generations of H. armigera, but not for the second generation. In a cotton and soybean/ peanut mix system, noncotton crops provided a natural refugia from the second- to fourth-generation H. armigera, but function of the refuge would closely depend on the proportion of Bt cotton. Consequently, it may be necessary to compensate the original mixed cropping patterns in different areas for delaying resistance development of H. armigera to Bt cotton.

Flavopiridol and Trastuzumab Synergistically Inhibit Proliferation of Breast Cancer Cells: Association with Selective Cooperative Inhibition of Cyclin D1-dependent Kinase and Akt Signaling Pathways

Molecular Cancer Therapeutics. Jul, 2002  |  Pubmed ID: 12479366

Cyclin D1 is essential for Neu-induced cell growth and is induced by growth factors through Ras-dependent and Ras-independent signaling pathways (1). Because flavopiridol, a novel flavanoid cyclin-cyclin-dependent kinase inhibitor, may function through Ras-dependent and/or -independent pathways, we hypothesized that treatment of breast cancer cells with inhibitors of Neu signaling and flavopiridol might synergize to inhibit proliferation. Human breast cancer cell lines, which express high levels of endogenous Neu receptor, were treated with the anti-Neu antibody, trastuzumab, together with flavopiridol and subject to MTT assay. Cell lines were assayed for alterations in cell cycle by fluorescence-activated cell sorter and signaling proteins by Western blot. Flavopiridol and trastuzumab synergistically inhibited DNA synthesis, cellular proliferation, and contact-dependent growth. Cytotoxic synergy was observed independent of the sequence of addition of the two drugs to cultured cells. In SKBR3 cells, a combination of trastuzumab and flavopiridol inhibited the Ras-MAPK-Akt pathway, decreased cyclin D1 abundance, and kinase activity to a greater extent than either drug alone. Compared with single-agent treatment, combination treatment selectively inhibited Akt and pRB phosphorylation. Cytotoxic synergy was observed with flavopiridol plus LY294002 (selective phosphatidylinositol 3-kinase inhibitor) but not with PD98059 (selective mitogen-activated protein kinase kinase 1 inhibitor) suggesting that Akt inhibition may be important in synergy. Zinc-induced overexpression of cyclin D1 in T-47D deltaMTcycD1 cells were more resistant to drug-induced cell death when compared with vector-transfected T-47D deltaMT cells. Cyclin D1 overexpression reverses drug treatment induced cell cycle arrest in SKBR3 cells. Flavopiridol and trastuzumab yield cytotoxic synergy in human breast cancer cells overexpressing Neu. Cyclin D1 overexpression results in combination drug resistance. In addition, inhibition of Akt may prove to be a useful therapeutic strategy in combination with flavopiridol.

IKKalpha Regulates Mitogenic Signaling Through Transcriptional Induction of Cyclin D1 Via Tcf

Molecular Biology of the Cell. Feb, 2003  |  Pubmed ID: 12589056

The Wnt/beta-catenin/Tcf and IkappaB/NF-kappaB cascades are independent pathways involved in cell cycle control, cellular differentiation, and inflammation. Constitutive Wnt/beta-catenin signaling occurs in certain cancers from mutation of components of the pathway and from activating growth factor receptors, including RON and MET. The resulting accumulation of cytoplasmic and nuclear beta-catenin interacts with the Tcf/LEF transcription factors to induce target genes. The IkappaB kinase complex (IKK) that phosphorylates IkappaB contains IKKalpha, IKKbeta, and IKKgamma. Here we show that the cyclin D1 gene functions as a point of convergence between the Wnt/beta-catenin and IkappaB pathways in mitogenic signaling. Mitogenic induction of G(1)-S phase progression and cyclin D1 expression was PI3K dependent, and cyclin D1(-/-) cells showed reduced PI3K-dependent S-phase entry. PI3K-dependent induction of cyclin D1 was blocked by inhibitors of PI3K/Akt/IkappaB/IKKalpha or beta-catenin signaling. A single Tcf site in the cyclin D1 promoter was required for induction by PI3K or IKKalpha. In IKKalpha(-/-) cells, mitogen-induced DNA synthesis, and expression of Tcf-responsive genes was reduced. Reintroduction of IKKalpha restored normal mitogen induction of cyclin D1 through a Tcf site. In IKKalpha(-/-) cells, beta-catenin phosphorylation was decreased and purified IKKalpha was sufficient for phosphorylation of beta-catenin through its N-terminus in vitro. Because IKKalpha but not IKKbeta induced cyclin D1 expression through Tcf activity, these studies indicate that the relative levels of IKKalpha and IKKbeta may alter their substrate and signaling specificities to regulate mitogen-induced DNA synthesis through distinct mechanisms.

Cyclin D1 Governs Adhesion and Motility of Macrophages

Molecular Biology of the Cell. May, 2003  |  Pubmed ID: 12802071

The cyclin D1 gene encodes the regulatory subunit of a holoenzyme that phosphorylates and inactivates the retinoblastoma protein, thereby promoting cell-cycle progression. Cyclin D1 is overexpressed in hematopoetic and epithelial malignancies correlating with poor prognosis and metastasis in several cancer types. Because tumor-associated macrophages have been shown to enhance malignant progression and metastasis, and cyclin D1-deficient mice are resistant to oncogene-induced malignancies, we investigated the function of cyclin D1-/- bone marrow-derived macrophages. Cyclin D1 deficiency increased focal complex formation at the site of substratum contact, and enhanced macrophage adhesion, yielding a flattened, circular morphology with reduced membrane ruffles. Migration in response to wounding, cytokine-mediated chemotaxis, and transendothelial cell migration of cyclin D1-/- bone marrow-derived macrophages were all substantially reduced. Thus, apart from proliferative and possible motility defects in the tumor cells themselves, the reduced motility and invasiveness of cyclin D1-/- tumor-associated macrophages may contribute to the tumor resistance of these mice.

The Role of Ink4a/Arf in ErbB2 Mammary Gland Tumorigenesis

Cancer Research. Jun, 2003  |  Pubmed ID: 12810676

Most human tumors display inactivation of the p53 and the p16(INK4)/pRb pathway. The Ink4a/alternative reading frame (ARF) locus encodes the p16(INK4a) and p14(ARF) (murine p19(ARF)) proteins. p16(INK4a) is deleted in 40-60% of breast cancer cell lines, and p16(INK4a) inactivation by DNA methylation occurs in < or =30% of human breast cancers. In mice genetically heterozygous for p16(INK4a) or Ink4a/Arf, predisposition to specific tumor types is enhanced. Ink4a/Arf(+/-) mice have increased E micro -Myc-induced lymphomagenesis and epidermal growth factor receptor-induced gliomagenesis. ErbB2 (epidermal growth factor receptor-related protein B2) is frequently overexpressed in human breast cancer and is sufficient for mammary tumorigenesis in vivo. We determined the role of heterozygosity at the Ink4a/Arf locus in ErbB2-induced mammary tumorigenesis. Compared with mouse mammary tumor virus-ErbB2 Ink4a/Arf(+/-) mice, mouse mammary tumor virus-ErbB2 Ink4a/Arf(wt) mammary tumors showed increased p16(INK4a), reduced Ki-67 expression, and reduced cyclin D1 protein but increased mammary tumor apoptosis with no significant change in the risk of developing mammary tumors. These studies demonstrate the contribution of Ink4a/Arf heterozygosity to tumor progression is tissue specific in vivo. In view of the important role of Ink4a/Arf in response to chemotherapy, these transgenic mice may provide a useful model for testing breast tumor therapies.

[HPLC Analysis of Main Flavonoid Chemicals and Their Spatio-temporal Dynamics in Bt Transgenic Cotton]

Ying Yong Sheng Tai Xue Bao = The Journal of Applied Ecology. Feb, 2003  |  Pubmed ID: 12827880

Flavonoids are important chemicals of resistance to pests in cotton plant. The main flavonoid chemicals and their spatio-temporal dynamics of content in Bt transgenic cotton were tested by HPLC. The results showed that the flavonoid chemicals of resistance to pests mainly including rutin, isoquercitrin and quercetin could be detected and quantitatively analyzed by HPLC. The contents of rutin, isoquercitrin and quercetin were the highest in petal, but lower in calyx, bract and cotton boll. Moreover, the total content of flavonoid chemicals in top leaf was much higher at developmental stage than at seedling stage. The content of each flavonoid chemicals of resistance to pests was different during different developmental stage and in different organs. It was indicated that different flavonoid chemicals played different roles in resistance to pests.

Cyclin D1 Repression of Peroxisome Proliferator-activated Receptor Gamma Expression and Transactivation

Molecular and Cellular Biology. Sep, 2003  |  Pubmed ID: 12917338

The cyclin D1 gene is overexpressed in human breast cancers and is required for oncogene-induced tumorigenesis. Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a nuclear receptor selectively activated by ligands of the thiazolidinedione class. PPAR gamma induces hepatic steatosis, and liganded PPAR gamma promotes adipocyte differentiation. Herein, cyclin D1 inhibited ligand-induced PPAR gamma function, transactivation, expression, and promoter activity. PPAR gamma transactivation induced by the ligand BRL49653 was inhibited by cyclin D1 through a pRB- and cdk-independent mechanism, requiring a region predicted to form an helix-loop-helix (HLH) structure. The cyclin D1 HLH region was also required for repression of the PPAR gamma ligand-binding domain linked to a heterologous DNA binding domain. Adipocyte differentiation by PPAR gamma-specific ligands (BRL49653, troglitazone) was enhanced in cyclin D1(-/-) fibroblasts and reversed by retroviral expression of cyclin D1. Homozygous deletion of the cyclin D1 gene, enhanced expression by PPAR gamma ligands of PPAR gamma and PPAR gamma-responsive genes, and cyclin D1(-/-) mice exhibit hepatic steatosis. Finally, reduction of cyclin D1 abundance in vivo using ponasterone-inducible cyclin D1 antisense transgenic mice, increased expression of PPAR gamma in vivo. The inhibition of PPAR gamma function by cyclin D1 is a new mechanism of signal transduction cross talk between PPAR gamma ligands and mitogenic signals that induce cyclin D1.

Efficacy of Transgenic Cotton Containing a Cry1Ac Gene from Bacillus Thuringiensis Against Helicoverpa Armigera (Lepidoptera: Noctuidae) in Northern China

Journal of Economic Entomology. Aug, 2003  |  Pubmed ID: 14503607

NuCOTN 33B, a Bt transgenic variety of upland cotton (Gossypium hirsutum L.) expressing the insecticidal protein Cry1Ac from Bacillus thuringiensis Berliner sp. kurstaki, was evaluated for resistance to Helicoverpa armigera (Hübner) during 1998-2000 in northern China. The results indicated that there was no significant difference in egg densities between NuCOTN 33B and three nontransgenic varieties (DP5415, Zhongmian12, and Shiyuan321) during the season, although the survival of larvae on NuCOTN 33B seemed significantly reduced. High larval densities observed on non-Bt cotton appeared in great contrast to the low larval populations observed on NuCOTN 33B plants during the seasons. In an environment without insecticide sprays, the annual ginned cotton yields in NuCOTN 33B plots, ranging from 1391.17 to 1511.35 kg/ha, were significantly higher than those in non-Bt cotton (340.34-359.58 kg/ha). These high levels of field efficacy for NuCOTN 33B against H. armigera in northern China may pave the way for reduced pesticide applications and an expansion of alternative pest-control strategies.

DACH1 Inhibits Transforming Growth Factor-beta Signaling Through Binding Smad4

The Journal of Biological Chemistry. Dec, 2003  |  Pubmed ID: 14525983

The vertebrate homologues of Drosophila dachsund, DACH1 and DACH2, have been implicated as important regulatory genes in development. DACH1 plays a role in retinal and pituitary precursor cell proliferation and DACH2 plays a specific role in myogenesis. DACH proteins contain a domain (DS domain) that is conserved with the proto-oncogenes Ski and Sno. Since the Ski/Sno proto-oncogenes repress AP-1 and SMAD signaling, we hypothesized that DACH1 might play a similar cellular function. Herein, DACH1 was found to be expressed in breast cancer cell lines and to inhibit transforming growth factor-beta (TGF-beta)-induced apoptosis. DACH1 repressed TGF-beta induction of AP-1 and Smad signaling in gene reporter assays and repressed endogenous TGF-beta-responsive genes by microarray analyses. DACH1 bound to endogenous NCoR and Smad4 in cultured cells and DACH1 co-localized with NCoR in nuclear dotlike structures. NCoR enhanced DACH1 repression, and the repression of TGF-beta-induced AP-1 or Smad signaling by DACH1 required the DACH1 DS domain. The DS domain of DACH was sufficient for NCoR binding at a Smad4-binding site. Smad4 was required for DACH1 repression of Smad signaling. In Smad4 null HTB-134 cells, DACH1 inhibited the activation of SBE-4 reporter activity induced by Smad2 or Smad3 only in the presence of Smad4. DACH1 participates in the negative regulation of TGF-beta signaling by interacting with NCoR and Smad4.

Geographic Variation in Susceptibility of Chilo Suppressalis (Lepidoptera: Pyralidae) to Bacillus Thuringiensis Toxins in China

Journal of Economic Entomology. Dec, 2003  |  Pubmed ID: 14977124

Geographic variation in the susceptibility of the striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), in China to Bacillus thuringiensis (Bt) insecticidal crystal proteins Cry1Ac and Cry1Ab was studied to establish baseline information for comparing the future response of populations with increased exposure to Bt products. Rice is the major host of C. suppressalis, and Bt rice ma) be released in China in the near future. Twelve populations of the pest were collected from the major rice-growing regions of China. LC50 estimates were determined for all populations for Cry1Ac and for eight populations for Cry1Ab. The bioassay results indicated that the range of LC50 in neonate larvae to Cry1Ac and Cry1Ab was from approximately 15 to approximately 157 mg (AI)/L and approximately 2 to approximately 34 mg (AI)/L, respectively. LC50 values were lower for Cry1Ab than for Cry1Ac, and there was a significant positive correlation between the two toxins tested.

The Androgen Receptor Acetylation Site Regulates CAMP and AKT but Not ERK-induced Activity

The Journal of Biological Chemistry. Jul, 2004  |  Pubmed ID: 15123687

The androgen receptor (AR) regulates ligand-dependent gene transcription upon binding specific DNA sequences. The AR conveys both trans-activation and trans-repression functions, which together contribute to prostate cellular growth, differentiation, and apoptosis. Like histone H3, the AR is post-translationally modified by both acetylation and phosphorylation. The histone acetyltransferase p300 transactivates the AR and directly acetylates the AR in vitro at a conserved motif. Point mutations of the AR acetylation motif that abrogate acetylation reduce trans-activation by p300 without affecting the trans-repression function of the AR. The current studies assessed the functional relationship between acetylation and phosphorylation of the AR. Herein trans-activation of the AR acetylation site mutants were enhanced by the p42/p44 MAPK pathway but were defective in regulation by protein kinase A (PKA) signaling. PKA inhibition augmented ARwt activity but not AR acetylation mutant gene reporter activity and association at an androgen response element in chromatin immunoprecipitation assays. Mutations of the lysine residues at the AR acetylation site reduced trichostatin A (TSA) responsiveness and ligand-induced phosphorylation of the AR. The AR acetylation site mutant formed ligand-induced phosphorylation-dependent isoforms with distinguishable characteristics from wild type AR as determined with two-dimensional electrophoresis. Conversely, point mutation of a subset of AR phosphorylation sites reduced trichostatin A responsiveness and trans-activation by histone acetyltransferases. Together these studies suggest that acetylation and phosphorylation of the AR are linked events and that the conserved AR lysine motif contributes to a select subset of pathways governing AR activity.

The Inhibitor of Cyclin-dependent Kinase 4a/alternative Reading Frame (INK4a/ARF) Locus Encoded Proteins P16INK4a and P19ARF Repress Cyclin D1 Transcription Through Distinct Cis Elements

Cancer Research. Jun, 2004  |  Pubmed ID: 15205322

The Ink4a/Arf locus encodes two structurally unrelated tumor suppressor proteins, p16(INK4a) and p14(ARF) (murine p19(ARF)). Invariant inactivation of either the p16(INK4a)-cyclin D/CDK-pRb pathway and/or p53-p14(ARF) pathway occurs in most human tumors. Cyclin D1 is frequently overexpressed in breast cancer cells contributing an alternate mechanism inactivating the p16(INK4a)/pRb pathway. Targeted overexpression of cyclin D1 to the mammary gland is sufficient for tumorigenesis, and cyclin D1-/- mice are resistant to Ras-induced mammary tumors. Recent studies suggest cyclin D1 and p16(INK4a) expression are reciprocal in human breast cancers. Herein, reciprocal regulation of cyclin D1 and p16(INK4a) was observed in tissues of mice mutant for the Ink4a/Arf locus. p16(INK4a) and p19(ARF) inhibited DNA synthesis in MCF7 cells. p16(INK4a) repressed cyclin D1 expression and transcription. Repression of cyclin D1 by p16(INK4a) occurred independently of the p16(INK4a)-cdk4-binding function and required a cAMP-response element/activating transcription factor-2-binding site. p19(ARF) repressed cyclin D1 through a novel distal cis-element at -1137, which bound p53 in chromatin-immunoprecipitation assays. Transcriptional repression of the cyclin D1 gene through distinct DNA sequences may contribute to the tumor suppressor function of the Ink4a/Arf locus.

Changes in Susceptibility to Conventional Insecticides of a Cry1Ac-selected Population of Helicoverpa Armigera (Hübner) (Lepidoptera: Noctuidae)

Pest Management Science. Jul, 2004  |  Pubmed ID: 15260299

The changes in the susceptibility of the cotton bollworm, Helicoverpa armigera (Hübner) to three insecticides (lambda-cyhalothrin, phoxim and endosulfan) commonly used for control of this pest in China were monitored by bioassays at various generations. The insects were originally collected from Bt cotton fields and selected with Cry1Ac over 44 generations. In comparison with a susceptible strain, the larval resistance of the Bt-selected populations to Cry1Ac toxin increased 106-fold. Simultaneously, the resistance levels to lambda-cyhalothrin, phoxim and endosulfan declined dramatically. The results indicated no positive cross-resistance between Cry1Ac toxin and the insecticides. Evidence of the lack of cross-resistance to three commonly used synthetic insecticides in our laboratory-derived Cry1Ac-resistant population may suggest that growers can confidently use these insecticides if and when resistance to Cry1Ac cotton does occur.

Regional Reversion of Insecticide Resistance in Helicoverpa Armigera (Lepidoptera: Noctuidae) is Associated with the Use of Bt Cotton in Northern China

Pest Management Science. May, 2005  |  Pubmed ID: 15643649

A continuous programme for monitoring resistance of Helicoverpa armigera (Hubner) to commonly used insecticides was undertaken for assessing the impact of Bt cotton planting on the evolution of the pest resistance from 1994 to 2002 in China. The bioassay results showed that the resistance levels per year in field populations of H armigera to lambda-cyhalothrin, phoxim and endosulfan decreased, respectively, to 9-15-fold, 11-14-fold and 6-8-fold in 2001-2002 from 197-262-fold, 52-74-fold and 18-38-fold before the cultivation of Bt cotton in 1997. This significant increase in susceptibility to insecticides is expected to result in a reduction in insecticide application for H armigera control in Bt cotton. It is concluded that Bt cotton is playing an important role in the long-term management of H armigera by increasing the potential for natural and chemical control of the pest.

Structural Determinants of the BRCA1 : Estrogen Receptor Interaction

Oncogene. Mar, 2005  |  Pubmed ID: 15674350

Previously, we showed that the BRCA1 protein interacts directly and functionally with estrogen receptor-alpha (ER-alpha), resulting in the inhibition of estradiol (E2)-stimulated ER-alpha transcriptional activity. The interaction sites were mapped to the N-terminus of BRCA1 (within amino acids (aa) 1-302) and the ligand-binding domain/activation function-2 (LBD/AF-2) region (within aa 282-420) of ER-alpha. In this study, we have further characterized the structure/function relationship for the BRCA1 : ER-alpha interaction. We found that the N-terminal RING domain (aa 20-64) is not required for the BRCA1 : ER-alpha interaction. We identified two separate contact points for ER-alpha, one within aa 1-100 and the other within aa 100-200 of BRCA1; and we showed that each of these BRCA1 peptides interacts with BRCA1 in vitro and in vivo. By using different fragments of the BRCA1 N-terminus, we found that aa 67-100 and 101-133 are required for the interaction with ER-alpha, but that aa 1-67 and 134-302 are dispensible. Previously, we showed that BRCA1 aa 1-302 does not inhibit E2-stimulated ER-alpha transcriptional activity but does bind to ER-alpha and acts as a dominant negative inhibitor of the full-length BRCA1 protein. Somewhat surprisingly, we found that BRCA1 aa 1-100 and BRCA1 aa 101-200 (but not aa 201-300) each inhibited ER-alpha activity, although not as efficiently as full-length BRCA1. Mutations within an HIV Rev-like nuclear export signal that resembles a nuclear receptor corepressor motif (aa 86-95) impaired the ability of both truncated (aa 1-100) and full-length (aa 1-1863) BRCA1 proteins to interact with and/or repress ER-alpha activity. Based on these findings, a partial BRCA1 : ER-alpha three-dimensional structure is proposed. The implications of these findings for understanding the BRCA1 : ER-alpha interaction are discussed.

Cyclin D1 Inhibits Peroxisome Proliferator-activated Receptor Gamma-mediated Adipogenesis Through Histone Deacetylase Recruitment

The Journal of Biological Chemistry. Apr, 2005  |  Pubmed ID: 15713663

The cyclin D1 gene encodes the labile serum-inducible regulatory subunit of a holoenzyme that phosphorylates and inactivates the retinoblastoma protein. Overexpression of cyclin D1 promotes cellular proliferation and normal physiological levels of cyclin D1 function to inhibit adipocyte differentiation in vivo. We have previously shown that cyclin D1 inhibits peroxisome proliferator-activated receptor (PPAR)gamma-dependent activity through a cyclin-dependent kinase- and retinoblastoma protein-binding-independent mechanism. In this study, we determined the molecular mechanism by which cyclin D1 regulated PPARgamma function. Herein, murine embryonic fibroblast (MEF) differentiation by PPARgamma ligand was associated with a reduction in histone deacetylase (HDAC1) activity. Cyclin D1-/- MEFs showed an increased propensity to undergo differentiation into adipocytes. Genetic deletion of cyclin D1 reduced HDAC1 activity. Reconstitution of cyclin D1 into the cyclin D1-/- MEFs increased HDAC1 activity and blocked PPARgamma-mediated adipogenesis. PPARgamma activity was enhanced in cyclin D1-/- cells. Reintroduction of cyclin D1 inhibited basal and ligand-induced PPARgamma activity and enhanced HDAC repression of PPARgamma activity. Cyclin D1 bound HDAC in vivo and preferentially physically associated with HDAC1, HDAC2, HDAC3, and HDAC5. Chromatin immunoprecipitation assay demonstrated that cyclin D1 enhanced recruitment of HDAC1 and HDAC3 and histone methyltransferase SUV39H1 to the PPAR response element of the lipoprotein lipase promoter and decreased acetylation of total histone H3 and histone H3 lysine 9. Collectively, these studies suggest an important role of cyclin D1 in regulation of PPARgamma-mediated adipocyte differentiation through recruitment of HDACs to regulate PPAR response element local chromatin structure and PPARgamma function.

Seasonal Expression Profiles of Insecticidal Protein and Control Efficacy Against Helicoverpa Armigera for Bt Cotton in the Yangtze River Valley of China

Journal of Economic Entomology. Feb, 2005  |  Pubmed ID: 15765683

Seasonal levels of Bacillus thuringiensis (Bt) insecticidal protein and its control efficacy against Helicoverpa armigera (Hübner) in Bt transgenic cotton GK19 (carrying a Cry1Ac/Cry1Ab fused gene) and BG1560 (carrying a Cry1Ac gene) were investigated in Tianmen County, Hubei Province, located in the Yangtze River valley of China, in 2001 and 2002. The results showed that the toxin content in Bt cotton changed significantly over time, and that the structure, growth stage, and variety were significant sources of variability. Generally, insecticidal protein levels were high during the early stages of cotton growth; they declined in mid-season, and rebounded in late season. On most dates sampled, the toxin contents in leaf, square, petal, and stamens (including nonovule pistil tissue) were much higher than those in ovule and boll. Compared with BG1560, the expression of Cry1Ac/Cry1Ab protein in GK19 was more variable during the whole growth period of cotton. The field evaluation on larval population dynamics of H. armigera in Bt and conventional cotton showed that the larval densities in BG1560 and GK19 fields decreased, respectively, 92.04 and 81.85% in 2001, and 96.84 and 91.80% in 2002.

A Study of Cytotoxic Synergy of UCN-01 and Flavopiridol in Syngeneic Pair of Cell Lines

Investigational New Drugs. Aug, 2005  |  Pubmed ID: 16012789

Flavopiridol and UCN-01 are two novel protein kinase inhibitors with diverse cellular effects that may complement each other with regards to induction of apoptosis. HeLa cells engineered to overexpress human survivin (HeLa-S) were at least approximately 4.8-fold resistant to UCN-01 relative to proliferation observed in control HeLa cells (HeLa-V). Flavopiridol cytotoxicity as measured using the MTT assay was unaffected in HeLa-S cells when compared with HeLa-V cells. Similarly, simultaneous treatment of HeLa-V cells with flavopiridol and UCN-01 for 72 hours did not result in synergistic inhibition of proliferation; however, in HeLa-S cells, this combination resulted in synergistic inhibition of cell proliferation. Flavopiridol and UCN-01 augmented apoptosis in HeLa-S cells (as compared with HeLa-V cells) as measured by caspase-3 cellular activity assay, DNA fragmentation and PARP cleavage by western blot. In HeLa-V and -S cells, combination treatment resulted in caspase-8 cleavage. Caspase-9 was expressed in HeLa-V cells; however, there was a marked reduction of caspase-9 content in HeLa-S cells only. Combination treatment resulted in a significant reduction in survivin abundance in HeLa-S and SKBR3-UR cells, but not in their respective parental lines. The synergy of Flavopiridol and UCN-01 are selectively toxic to survivin-overexpressing cell lines and the mechanism of toxicity involves caspase-dependent cell death.

Gene Cloning and Expression of Cadherin in Midgut of Helicoverpa Armigera and Its Cry1A Binding Region

Science in China. Series C, Life Sciences. Aug, 2005  |  Pubmed ID: 16248428

Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin-like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect's resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence, suggesting that the decrease of expression quantity of BtR-harm is one of the main reasons for H. armigera resistance to Cry1Ac.

Regulation of AlphaA-crystallin Via Pax6, C-Maf, CREB and a Broad Domain of Lens-specific Chromatin

The EMBO Journal. May, 2006  |  Pubmed ID: 16675956

Pax6 and c-Maf regulate multiple stages of mammalian lens development. Here, we identified novel distal control regions (DCRs) of the alphaA-crystallin gene, a marker of lens fiber cell differentiation induced by FGF-signaling. DCR1 stimulated reporter gene expression in primary lens explants treated with FGF2 linking FGF-signaling with alphaA-crystallin synthesis. A DCR1/alphaA-crystallin promoter (including DCR2) coupled with EGFP virtually recapitulated the expression pattern of alphaA-crystallin in lens epithelium and fibers. In contrast, the DCR3/alphaA/EGFP reporter was expressed only in 'late' lens fibers. Chromatin immunoprecipitations showed binding of Pax6 to DCR1 and the alphaA-crystallin promoter in lens chromatin and demonstrated that high levels of alphaA-crystallin expression correlate with increased binding of c-Maf and CREB to the promoter and of CREB to DCR3, a broad domain of histone H3K9-hyperacetylation extending from DCR1 to DCR3, and increased abundance of chromatin remodeling enzymes Brg1 and Snf2h at the alphaA-crystallin locus. Our data demonstrate a novel mechanism of Pax6, c-Maf and CREB function, through regulation of chromatin-remodeling enzymes, and suggest a multistage model for the activation of alphaA-crystallin during lens differentiation.

Resistance Monitoring of Helicoverpa Armigera (Lepidoptera: Noctuidae) to Bt Insecticidal Protein During 2001-2004 in China

Journal of Economic Entomology. Jun, 2006  |  Pubmed ID: 16813327

Susceptibility of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) field populations to the CrylAc toxin of Bacillus thuringiensis Berliner (Bt) were monitored from 1997 to 2004 in China. During 2001-2004, 53 strains from the Bt cotton planting region were sampled. The range of concentration producing 50% inhibition of larval development to third instar (IC50) values among different populations in 2001, 2002, 2003, and 2004 was 0.014-0.046, 0.010-0.062, 0.005-0.062, and 0.005-0.035 microg/ml, respectively. Diagnostic concentration studies (IC99) showed that the percentage of individuals reaching third instar ranged from 0 to 9.09%, with only four of the 53 tested populations showing values above 0%. Considering these data, it was determined that the susceptibility to CrylAc of the field populations sampled was not different from the baseline in 1997, and no movement toward resistance among H. armigera populations was apparent.

Expression Profiling Identifies Altered Expression of Genes That Contribute to the Inhibition of Transforming Growth Factor-beta Signaling in Ovarian Cancer

Cancer Research. Sep, 2006  |  Pubmed ID: 16951150

Ovarian cancer is resistant to the antiproliferative effects of transforming growth factor-beta (TGF-beta); however, the mechanism of this resistance remains unclear. We used oligonucleotide arrays to profile 37 undissected, 68 microdissected advanced-stage, and 14 microdissected early-stage papillary serous cancers to identify signaling pathways involved in ovarian cancer. A total of seven genes involved in TGF-beta signaling were identified that had altered expression >1.5-fold (P < 0.001) in the ovarian cancer specimens compared with normal ovarian surface epithelium. The expression of these genes was coordinately altered: genes that inhibit TGF-beta signaling (DACH1, BMP7, and EVI1) were up-regulated in advanced-stage ovarian cancers and, conversely, genes that enhance TGF-beta signaling (PCAF, TFE3, TGFBRII, and SMAD4) were down-regulated compared with the normal samples. The microarray data for DACH1 and EVI1 were validated using quantitative real-time PCR on 22 microdissected ovarian cancer specimens. The EVI1 gene locus was amplified in 43% of the tumors, and there was a significant correlation (P = 0.029) between gene copy number and EVI1 gene expression. No amplification at the DACH1 locus was found in any of the samples. DACH1 and EVI1 inhibited TGF-beta signaling in immortalized normal ovarian epithelial cells, and a dominant-negative DACH1, DACH1-Delta DS, partially restored signaling in an ovarian cancer cell line resistant to TGF-beta. These results suggest that altered expression of these genes is responsible for disrupted TGF-beta signaling in ovarian cancer and they may be useful as new and novel therapeutic targets for ovarian cancer.

P27Kip1 Repression of ErbB2-induced Mammary Tumor Growth in Transgenic Mice Involves Skp2 and Wnt/beta-catenin Signaling

Cancer Research. Sep, 2006  |  Pubmed ID: 16951165

Expression of the cyclin-dependent kinase (Cdk) inhibitor (p27(Kip1)) is frequently reduced in human tumors, often correlating with poor prognosis. p27(Kip1) functions as a haploinsufficient tumor suppressor; however, the mechanism by which one allele of p27(Kip1) regulates oncogenic signaling in vivo is not well understood. We therefore investigated the mechanisms by which p27(Kip1) inhibits mammary tumor onset. Using the common background strain of FVB, p27(Kip1) heterozygosity (p27(+/-)) accelerated ErbB2-induced mammary tumorigenesis. We conducted microarray analyses of mammary tumors developing in mice with genetic haploinsufficiency for p27(Kip1) expressing a mammary-targeted ErbB2 oncogene. Global gene expression profiling and Western blot analysis of ErbB2/p27(+/-) tumors showed that the loss of p27(Kip1) induced genes promoting lymphangiogenesis, cellular proliferation, and collaborative oncogenic signaling (Wnt/beta-catenin/Tcf, Cdc25a, Smad7, and Skp2). Skp2 expression was induced by ErbB2 and repressed by p27(Kip1). Degradation of p27(Kip1) involves an SCF-type E3 ubiquitin ligase, including Skp2. The Skp2 component of the SCF(SKP2) complex that degrades p27(Kip1) was increased in ErbB2 tumors correlating with earlier tumor onset. In both murine and human ErbB2-overexpressing breast cancers, p27(Kip1) levels correlated inversely with Skp2. p27(Kip1) haploinsufficiency activated Wnt/beta-catenin/hedgehog signaling. Reintroduction of p27(Kip1) inhibited beta-catenin induction of Tcf-responsive genes (Siamosis, c-Myc, and Smad7). p27(Kip1) is haploinsufficient for ErbB2 mammary tumor suppression in vivo and functions to repress collaborative oncogenic signals including Skp2 and Wnt/beta-catenin signaling.

DACH1 is a Cell Fate Determination Factor That Inhibits Cyclin D1 and Breast Tumor Growth

Molecular and Cellular Biology. Oct, 2006  |  Pubmed ID: 16980615

Obstacles to the expansion of cells with proliferative potential include the induction of cell death, telomere-based senescence, and the pRb and p53 tumor suppressors. Not infrequently, the molecular pathways regulating oncogenesis recapitulate aberrations of processes governing embryogenesis. The genetic network, consisting of the dachshund (dac), eyes absent (eya), eyeless, and sine oculis (so) genes, regulates cell fate determination in metazoans, with dac serving as a cointegrator through a So DNA-binding factor. Here, DACH1 inhibited oncogene-mediated breast oncogenesis, blocking breast cancer epithelial cell DNA synthesis, colony formation, growth in Matrigel, and tumor growth in mice. Genetic deletion studies demonstrated a requirement for cyclin D1 in DACH1-mediated inhibition of DNA synthesis. DACH1 repressed cyclin D1 through a novel mechanism via a c-Jun DNA-binding partner, requiring the DACH1 alpha-helical DS domain which recruits corepressors to the local chromatin. Analysis of over 2,000 patients demonstrated increased nuclear DACH1 expression correlated inversely with cellular mitosis and predicted improved breast cancer patient survival. The cell fate determination factor, DACH1, arrests breast tumor proliferation and growth in vivo providing a new mechanistic and potential therapeutic insight into this common disease.

Cell Fate Determination Factor DACH1 Inhibits C-Jun-induced Contact-independent Growth

Molecular Biology of the Cell. Mar, 2007  |  Pubmed ID: 17182846

The cell fate determination factor DACH1 plays a key role in cellular differentiation in metazoans. DACH1 is engaged in multiple context-dependent complexes that activate or repress transcription. DACH1 can be recruited to DNA via the Six1/Eya bipartite transcription (DNA binding/coactivator) complex. c-Jun is a critical component of the activator protein (AP)-1 transcription factor complex and can promote contact-independent growth. Herein, DACH1 inhibited c-Jun-induced DNA synthesis and cellular proliferation. Excision of c-Jun with Cre recombinase, in c-jun(f1/f1) 3T3 cells, abrogated DACH1-mediated inhibition of DNA synthesis. c-Jun expression rescued DACH1-mediated inhibition of cellular proliferation. DACH1 inhibited induction of c-Jun by physiological stimuli and repressed c-jun target genes (cyclin A, beta-PAK, and stathmin). DACH1 bound c-Jun and inhibited AP-1 transcriptional activity. c-jun and c-fos were transcriptionally repressed by DACH1, requiring the conserved N-terminal (dac and ski/sno [DS]) domain. c-fos transcriptional repression by DACH1 requires the SRF site of the c-fos promoter. DACH1 inhibited c-Jun transactivation through the delta domain of c-Jun. DACH1 coprecipitated the histone deacetylase proteins (HDAC1, HDAC2, and NCoR), providing a mechanism by which DACH1 represses c-Jun activity through the conserved delta domain. An oncogenic v-Jun deleted of the delta domain was resistant to DACH1 repression. Collectively, these studies demonstrate a novel mechanism by which DACH1 blocks c-Jun-mediated contact-independent growth through repressing the c-Jun delta domain.

Efficacy of Transgenic Rice Expressing Cry1Ac and CpTI Against the Rice Leaffolder, Cnaphalocrocis Medinalis (Guenée)

Journal of Invertebrate Pathology. Sep, 2007  |  Pubmed ID: 17445827

Detached leaf bioassays, open field tests and cage tests were conducted to evaluate the control efficacy of two transgenic rice lines, expressing Cry1Ac and CpTI, against Cnaphalocrocis medinalis (Guenée) during 2005-2006 in Fuzhou, China. Bioassay results showed that cumulative feeding areas of C. medinalis on transgenic lines were significantly lower than that on control rice lines at different developmental stages. The corrected mortalities at 96 h after infestation on transgenic lines during six rice growth stages were >90% and 100% during experiments conducted in 2005 and 2006, respectively. In the open field test, there was no significant difference in egg density between transgenic and control lines during early days of infestation, but significant differences were detected in late season, due to serious damage on control lines. Larval densities on control lines were significantly higher than the low larval populations observed on transgenic lines during both seasons. The percentages of plants with folded leaves and percentages of folded leaves on transgenic lines were significantly lower than that on control lines with and without insecticide applications, during the entire season. In cage tests the cumulative numbers of C. medinalis adults derived from transgenic lines were significantly lower than that from control lines with and without insecticide treatments. The high level of efficacy of the two transgenic rice lines against C. medinalis may provide an important basis for reduced insecticide applications, an expansion of alternative pest-control strategies and insect resistance management of Bt rice in the future.

Monitoring and Management Strategy for Helicoverpa Armigera Resistance to Bt Cotton in China

Journal of Invertebrate Pathology. Jul, 2007  |  Pubmed ID: 17467730

The cotton bollworm, Helicoverpa armigera, is one of the most important insect pests in cotton growing regions of China. Transgenic cotton that expresses a gene derived from the bacterium Bacillus thuringiensis (Bt) has been deployed for combating cotton bollworm since 1997. Natural refugees derived from the mixed planting system consisting of cotton, corn, soybean, vegetables, peanut and others on single-family farms of a small scale were used for delaying the evolution of resistance to Bt cotton. Susceptibility of H. armigera field populations to the Bt insecticidal protein Cry1Ac was monitored from 1997 to 2006. The results indicate that the field populations are still susceptible to Cry1Ac, and monitoring indication no apparent shifts in susceptibility in field populations of this important pest.

Cross-resistance Studies of Cry1Ac-resistant Strains of Helicoverpa Armigera (Lepidoptera: Noctuidae) to Cry2Ab

Journal of Economic Entomology. Jun, 2007  |  Pubmed ID: 17598555

Bioassays and binding tests between Cry toxins (CrylAa, CrylAb, CrylAc, and Cry2Ab) and brush-border membrane vesicles (BBMVs) from larvae of a Bacillus thurningiensis (Bt)-susceptible (96S) and two CrylAc-resistant strains (BtR and LFR10) were conducted for investigating cross-resistance of CrylAc-resistant strains to Cry2Ab in Helicoverpa armigera (Hiibner) (Lepidoptera: Noctuidae). The resistance ratio (RR) values of the BtR and LFR10 strains to CrylAc and Cry2Ab were 2,971- and 1.1-fold and 253- and 1.0-fold, respectively, indicating that there was no cross-resistance to Cry2Ab. The binding experiments between Cry toxins and BBMVs from BtR, LFR10, and 96S larvae showed that all of the toxins could bind with these BBMVs, but the Cry2Ab could not displace 125I labeled CrylAc and CrylAb. The same results were observed in reciprocal binding tests, demonstrating that CrylA and Cry2Ab had different binding sites in H. armigera and providing a potential mechanism for the lack of cross-resistance between CrylA and Cry2Ab toxins. These results suggest that the transgenic cotton, Gossypium hirsutum L., expressing CrylAc and Cry2Ab genes may be deployed for management of CrylAc resistant H. armigera.

Population Dynamics of Aphis Glycines (Homoptera: Aphididae) and Impact of Natural Enemies in Northern China

Environmental Entomology. Aug, 2007  |  Pubmed ID: 17716474

Field surveys of soybean aphid, Aphis glycines Matsumura, and its natural enemies, as well as natural enemy exclosure experiments, were conducted during 2003 and 2004 in soybean fields near Langfang, China. In 2003, aphid density increased six-fold during 12 d in July from 66+/-12 per 10 plants to a seasonal peak of 401+/-79 per 10 plants. Aphid density remained high for another 10 d and declined during late July and early August. In 2004, aphid density increased 29-fold during 13 d in July from 14+/-2 per 10 plants to a seasonal peak of 375+/-30 per 10 plants. Unlike 2003, aphid density remained relatively high during late July and August, peaking again at 296+/-31 per 10 plants on 24 August. In both years, aphid density remained below economic injury level and seemed to be limited by natural enemies. Exclosure of natural enemies led to increases in A. glycines density in 2003 and 2004. In 2003, peak aphid densities in large- and medium-mesh cages were three- and seven-fold higher, respectively, than densities on uncaged plants. In 2004, peak aphid densities in large- and medium-mesh cages were 2-fold and 30-fold higher, respectively, than densities on uncaged plants in one experiment. In another experiment, peak aphid densities in large-, medium-, and small-mesh cages were 8-fold, 28-fold, and 68-fold higher, respectively, than densities on uncaged plants. Both predators and parasitoids were important in limiting aphid density. We compare our results with those from North America and discuss implications for biological control.

Flight Potential of Lygus Lucorum (Meyer-Dür) (Heteroptera: Miridae)

Environmental Entomology. Oct, 2007  |  Pubmed ID: 18284721

Lygus lucorum (Meyer-Dür) is a key pest of Bt cotton in China. This study reports on its flight potential examined by a flight-mill system. We found that 10-d-old mated females engaged in flight over the greatest distance (40.1 +/- 5.2 km) and duration (7.7 +/- 1.0 h) in 24-h flight assays in relation to age, sex, and mating status. Optimum temperature for flight was 20 degrees C, and optimum relative humidity was 75% RH. Flight potential of 10-d-old mated females under the optimum conditions (20 degrees C and 75% RH) was tested continuously for 48 h. Results showed that the flight distance amounted to 67.3 +/- 9.7 km, with a maximum distance of 151.3 km. This study shows that L. lucorum has the potential to undertake long-distance flight. The information will help in the development of the forecast and management of L. lucorum.

Degradation of Cry1Ac Protein Within Transgenic Bacillus Thuringiensis Rice Tissues Under Field and Laboratory Conditions

Environmental Entomology. Oct, 2007  |  Pubmed ID: 18284753

To clarify the environmental fate of the Cry1Ac protein from Bacillus thuringiensis subsp. kurstaki (Bt) contained in transgenic rice plant stubble after harvest, degradation was monitored under field conditions using an enzyme-linked immunosorbent assay. In stalks, Cry1Ac protein concentration decreased rapidly to 50% of the initial amount during the first month after harvest; subsequently, the degradation decreased gradually reaching 21.3% when the experiment was terminated after 7 mo. A similar degradation pattern of the Cry1Ac protein was observed in rice roots. However, when the temperature increased in April of the following spring, protein degradation resumed, and no protein could be detected by the end of the experiment. In addition, a laboratory experiment was conducted to study the persistence of Cry1Ac protein released from rice tissue in water and paddy soil. The protein released from leaves degraded rapidly in paddy soil under flooded conditions during the first 20 d and plateaued until the termination of this trial at 135 d, when 15.3% of the initial amount was still detectable. In water, the Cry1Ac protein degraded more slowly than in soil but never entered a relatively stable phase as in soil. The degradation rate of Cry1Ac protein was significantly faster in nonsterile water than in sterile water. These results indicate that the soil environment can increase the degradation of Bt protein contained in plant residues. Therefore, plowing a field immediately after harvest could be an effective method for decreasing the persistence of Bt protein in transgenic rice fields.

DACH1 Negatively Regulates the Human RANK Ligand Gene Expression in Stromal/preosteoblast Cells

Journal of Cellular Biochemistry. Apr, 2008  |  Pubmed ID: 17891780

Receptor activator of NF-kappaB ligand (RANKL) is a critical osteoclastogenic factor that is expressed on bone marrow stromal/preosteoblast cells. Most bone resorption stimuli induce osteoclast formation by modulating RANKL expression in these cells. However, little is known about the mechanisms regulating RANKL gene expression. We recently reported that heat shock factor-2 (HSF-2) is a downstream target for FGF-2 signaling to enhance RANKL gene transcription in marrow stromal/preosteoblast cells. In this study, we show that DACH1 (human homologue of Drosophila dachshund gene) negatively regulates RANKL gene expression and suppresses FGF-2-enhanced RANKL gene expression in these cells. DACH1 contains a conserved dachshund domain (DS) in the N-terminal region, which interacts with the nuclear co-repressor (NCoR) to repress gene expression. Co-expression of DACH1 with hRANKL promoter-luciferase reporter plasmid in normal human bone marrow-derived stromal cells significantly decreased (3.3-fold) FGF-2-stimulated hRANKL gene promoter activity. Deletion of DS domain abolished DACH1 inhibition of FGF-2-enhanced RANKL gene promoter activity. Western blot analysis confirmed that DACH1 suppressed FGF-2-stimulated RANKL expression in marrow stromal/preosteoblast cells. We show HSF-2 co-immune precipitated with DACH1 and that FGF-2 stimulation significantly increased (2.7-fold) HSF-2 binding to DACH1. Confocal microscopy analysis further demonstrated that FGF-2 promotes HSF-2 nuclear transport and co-localization with DACH1 in marrow stromal cells. Co-expression of NCoR with DACH1 significantly decreased (5.3-fold) and siRNA suppression of NCoR in DACH1 co-transfected cells increased (3.6-fold) RANKL promoter activity. Furthermore, DACH1 co-expression with NCoR significantly decreased (7.5-fold) RANKL mRNA expression in marrow stromal cells. Collectively, these studies indicate that NCoR participates in DACH1 repression of RANKL gene expression in marrow stromal/preosteoblast cells. Thus, DACH1 plays an important role in negative regulation of RANKL gene expression in marrow stromal/preosteoblast cells in the bone microenvironment.

Disruption of C-Jun Reduces Cellular Migration and Invasion Through Inhibition of C-Src and Hyperactivation of ROCK II Kinase

Molecular Biology of the Cell. Apr, 2008  |  Pubmed ID: 18216279

The spread of metastatic tumors to different organs is associated with poor prognosis. The metastatic process requires migration and cellular invasion. The protooncogene c-jun encodes the founding member of the activator protein-1 family and is required for cellular proliferation and DNA synthesis in response to oncogenic signals and plays an essential role in chemical carcinogenesis. The role of c-Jun in cellular invasion remains to be defined. Genetic deletion of c-Jun in transgenic mice is embryonic lethal; therefore, transgenic mice encoding a c-Jun gene flanked by LoxP sites (c-jun(f/f)) were used. c-jun gene deletion reduced c-Src expression, hyperactivated ROCK II signaling, and reduced cellular polarity, migration, and invasiveness. c-Jun increased c-Src mRNA abundance and c-Src promoter activity involving an AP-1 site in the c-Src promoter. Transduction of c-jun(-/-) cells with either c-Jun or c-Src retroviral expression systems restored the defective cellular migration of c-jun(-/-) cells. As c-Src is a critical component of pathways regulating proliferation, survival, and metastasis, the induction of c-Src abundance, by c-Jun, provides a novel mechanism of cooperative signaling in cellular invasion.

Flight Performance of the Soybean Aphid, Aphis Glycines (Hemiptera: Aphididae) Under Different Temperature and Humidity Regimens

Environmental Entomology. Apr, 2008  |  Pubmed ID: 18419900

The soybean aphid, Aphis glycines (Matsumura), is native to eastern Asia and has recently invaded North America, where it is currently the most important insect pest of soybeans. The soybean aphid has spread rapidly within North America, presumably through a combination of active and passive (wind-aided) flight. Here, we studied the active flight potential of A. glycines under a range of environmental conditions using an aphid flight mill. Winged (alate) A. glycines were tested on a specially designed 32-channel, computer-monitored flight mill system. Aphids that were 12-24 h old exhibited the strongest flight behavior, with average flight durations of 3.3-4.1 h, which represented flight distances of 4.6-5.1 km. After the age of 72 h, A. glycines flight performance rapidly declined. The optimum temperature range for flight was 16-28 degrees C, whereas optimum relative humidity was 75%. Our findings show that A. glycines posseses a fairly strong active flight aptitude (ability and inclination) and point to the possibility of flight initiation under a broad range of environmental conditions. These results have the potential to aid forecasting and management protocols for A. glycines at the landscape level.

Dachshund Inhibits Oncogene-induced Breast Cancer Cellular Migration and Invasion Through Suppression of Interleukin-8

Proceedings of the National Academy of Sciences of the United States of America. May, 2008  |  Pubmed ID: 18467491

Oncogene-mediated signaling to the host environment induces a subset of cytokines and chemokines. The Drosophila Dac gene promotes migration of the morphogenetic furrow during eye development. Expression of the cell-fate determination factor Dachshund (DACH1) was lost in poor prognosis invasive breast cancer. Mouse embryo fibroblasts derived from Dach1(-/-) mice demonstrated endogenous Dach1 constitutively represses cellular migration. DACH1 inhibited cellular migration and invasion of oncogene (Ras, Myc, ErbB2, c-Raf)-transformed human breast epithelial cells. An unbiased proteomic analysis identified and immunoneutralizing antibody and reconstitution experiments demonstrated IL-8 is a critical target of DACH1 mediating breast cancer cellular migration and metastasis in vivo. DACH1 bound the endogenous IL-8 promoter in ChIP assays and repressed the IL-8 promoter through the AP-1 and NF-kappaB binding sites. Collectively, our data identify a pathway by which an endogenous cell-fate determination factor blocks oncogene-dependent tumor metastasis via a key heterotypic mediator.

A Cyclin D1/microRNA 17/20 Regulatory Feedback Loop in Control of Breast Cancer Cell Proliferation

The Journal of Cell Biology. Aug, 2008  |  Pubmed ID: 18695042

Decreased expression of specific microRNAs (miRNAs) occurs in human tumors, which suggests a function for miRNAs in tumor suppression. Herein, levels of the miR-17-5p/miR-20a miRNA cluster were inversely correlated to cyclin D1 abundance in human breast tumors and cell lines. MiR-17/20 suppressed breast cancer cell proliferation and tumor colony formation by negatively regulating cyclin D1 translation via a conserved 3' untranslated region miRNA-binding site, thereby inhibiting serum-induced S phase entry. The cell cycle effect of miR-17/20 was abrogated by cyclin D1 siRNA and in cyclin D1-deficient breast cancer cells. Mammary epithelial cell-targeted cyclin D1 expression induced miR-17-5p and miR-20a expression in vivo, and cyclin D1 bound the miR-17/20 cluster promoter regulatory region. In summary, these studies identify a novel cyclin D1/miR-17/20 regulatory feedback loop through which cyclin D1 induces miR-17-5p/miR-20a. In turn, miR-17/20 limits the proliferative function of cyclin D1, thus linking expression of a specific miRNA cluster to the regulation of oncogenesis.

Population Dynamics of Spodoptera Litura (Lepidoptera: Noctuidae) on Bt Cotton in the Yangtze River Valley of China

Environmental Entomology. Aug, 2008  |  Pubmed ID: 18801271

Genetically modified cotton that produces a crystalline protein from Bacillus thuringiensis subsp. kurstaki (Berliner) (Bt) has been widely deployed to manage lepidopteran insect pests in cotton growing areas worldwide. However, susceptibility of different insect species to Bt protein varies, which may affect lepidopteran pest populations in the field. Studies on effects of two transgenic cotton lines (BG1560 and GK19) carrying a Cry1A gene on common cutworm Spodoptera litura F. (Lepidoptera: Noctuidae), were conducted during 2002-2005 in the cotton planting region of the Yangtze River valley of China. Results showed that common cutworm larvae had low susceptibility to Bt cotton. There was no significant difference in larval population densities in conventional and Bt cotton fields. However, the larval populations of the insect on conventional plants treated with chemical insecticides for control of target pest of Bt cotton were significantly lower than that in Bt cotton fields. These results indicated that the common cutworm was the potential to become a major and alarming pest in Bt cotton fields, and therefore efforts to develop an effective alternative management strategy are needed.

Population Dynamics of Sesamia Inferens on Transgenic Rice Expressing Cry1Ac and CpTI in Southern China

Environmental Entomology. Oct, 2008  |  Pubmed ID: 19036217

Genetically modified insect-resistant rice lines containing the cry1Ac gene from Bacillus thuringiensis (Bt) or the CpTI (cowpea trypsin inhibitor) gene developed for the management of lepidopterous pests are highly resistant to the major target pests, Chilo suppressalis (Walker), Cnaphalocrocis medinalis (Guenée), and Scirpophaga incertulas (Walker), in the main rice-growing areas of China. However, the effects of these transgenic lines on Sesamia inferens (Walker), an important lepidopterous rice pest, are currently unknown. Because different insect species have varying susceptibility to Bt insecticidal proteins that may affect population dynamics, research into the effects of these transgenic rice lines on the population dynamics of S. inferens was conducted in Fuzhou, southern China, in 2005 and 2006. The results of laboratory, field cage, and field plot experiments show that S. inferens has comparatively high susceptibility to the transgenic line during the early growing season, with significant differences observed in larval density and infestation levels between transgenic and control lines. Because of a decrease in Cry1Ac levels in the plant as it ages, the transgenic line provided only a low potential for population suppression late in the growing season. There is a correlation between the changing expression of Cry1Ac and the impact of transgenic rice on the population dynamics of S. inferens during the season. These results indicate that S. inferens may become a major pest in fields of prospective commercially released transgenic rice, and more attention should be paid to developing an effective alternative management strategy.

Lethal and Sublethal Effects of Endosulfan on Apolygus Lucorum (Hemiptera: Miridae)

Journal of Economic Entomology. Dec, 2008  |  Pubmed ID: 19133460

Various pesticides directly impact insect pest populations through acute toxicity (lethal) effects, but they also cause a range of indirect (sublethal) effects on pest physiology or behavior. In this study, we describe both lethal and sublethal effects of endosulfan on the survival, development, and fecundity of Apolygus lucorum Meyer-Dür (Hemiptera: Miridae), an important pest of Bacillus thuringiensis (Bt) cotton, Gossypium hirsutum L., in China. We quantify pesticide effects on adult mirids as well as on subsequent (F1, F2) generations. Toxicity bioassays determined the median lethal concentration (LC50) for adult A. lucorum as 0.58 ppm. To evaluate the sublethal effect of endosulfan, we used LC20 and LC40 concentrations at 0.30 ppm and 0.48 ppm, respectively. Sublethal concentrations of endosulfan significantly reduced longevity and fecundity of female adults, but slightly affected male longevity. We also found sublethal effects on the F1 generation, through a reduction of egg hatching rate, nymphal mortality, adult molting rate, and adult fecundity, and on the F2 generation, through a decrease of egg hatching rate. These results show that endosulfan has far greater effects on A. lucorum than estimated by acute toxicity tests, and the effects of this pesticide on surviving mirids can be comparatively long-lived. This pesticide likely causes severe disruptions of A. lucorum population dynamics, with single endosulfan application, invoking a great reduction of population growth of resident mirid populations. We discuss implications of this work for control programs for A. lucorum on Bt cotton.

Field-testing of Synthetic Herbivore-induced Plant Volatiles As Attractants for Beneficial Insects

Environmental Entomology. Dec, 2008  |  Pubmed ID: 19161683

Seven synthetic herbivore-induced plant volatiles (HIPVs) and a mixture of nonanal + (Z)-3-hexen-1-ol were field tested for their ability to attract beneficial insects in an open cotton field. Eleven species of the main natural enemies of insect pests in cotton fields were studied. Significantly greater numbers of the ladybird beetle Coccinella septempunctata were trapped on (Z)-3-hexenyl acetate-baited cards than on others that were HIPV baited or the control cards. Erigonidium graminicolum was attracted to traps baited with nonanal, (Z)-3-hexenyl acetate, and methyl salicylate (MeSA). The predatory bug Deraeocoris punctulatus was only attracted to traps baited with octanal. The predatory bug Orius similis responded to traps baited with 3,7-dimethyl,1,3,6-octatriene, nonanal, (Z)-3-hexenyl acetate, nonanal + (Z)-3-hexen-1-ol, and MeSA. Dimethyl octatriene, nonanal + (Z)-3-hexen-1-ol, and octanal significantly attracted the syrphid fly Paragus quadrifasciatus. The ladybird beetle Propylaea japonica, the green lacewing Chrysopa sinica, the bigeyed bug Geocoris pallidipennis, the syrphid fly Epistrophe balteata, and the parasitic wasp Campoletis chlorideae did not respond to any of the HIPVs tested. These results are discussed with regard to the potential of HIPVs as tools for recruiting natural enemies into cotton fields.

Seasonal Migration of Helicoverpa Armigera (Lepidoptera: Noctuidae) over the Bohai Sea

Journal of Economic Entomology. Feb, 2009  |  Pubmed ID: 19253623

The seasonal migration of the Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) over the Bohai Sea was observed with a searchlight trap and an entomological radar located on a small island in the center of the sea, and through a network of light-traps around the Bohai region. The H. armigera moths were observed to migrate over the sea at least as early as May and light trapping through a network suggested migration might start as early as April, as soon as the moths had emerged from overwintering pupae. H. armigera moths migrated toward the north in southerly winds during spring and summer, and returned south on nights with northerly winds, or at altitudes where the wind was northerly, during fall. The passage of a weather front (cold or warm) or trough at approximately 1700 hours provokes migration of H. armigera over the sea. The H. armigera generally flew at altitudes of below 1,500 m above sea level (asl) with layer concentrations at 200-500 m asl, where the wind direction, wind speed, and temperature were optimum. During fall migration, H. armigera tended to orient toward the southwest and was able to compensate for the wind drift by turning clockwise when the downwind direction was < 225 degrees but counterclockwise when it was > 225 degrees. The displacement speed measured with the radar was 24-41 km/h, the duration of flight was 8-11 h and the maximum migration rate was 1,894 moths per km per h.

Trade-offs Between Flight and Fecundity in the Soybean Aphid (Hemiptera: Aphididae)

Journal of Economic Entomology. Feb, 2009  |  Pubmed ID: 19253628

The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is native to eastern Asia and was accidentally introduced into North America in 2000. Within a few years, it was found throughout the U.S. and Canadian soybean-growing regions. The spread of A. glycines in North America is partly ascribed to its great flight capacity. We conducted direct measurements of flight performance of winged A. glycines adults and correlated this to their subsequent longevity and fecundity to determine whether there are trade-offs between flight and fecundity. We also estimated fecundity and development time of the offspring of females that had flown varying distances to examine potential maternal effects linked to flight. The experiment was conducted using a specifically designed aphid flight mill system in which winged aphids were tethered by their abdomens and flight duration, distance and speed were quantified. Fecundity, longevity and reproductive periods of 12-h-old A. glycines alates that had engaged in > 0.5 km long flights were significantly lower than those of < 0.5-km individuals. The offspring of alates with flight experiences of > 1.5 km also had lower fecundity than those produced by individuals that had engaged in flights < 1.5 km. Our results are therefore consistent both with direct trade-offs between flight and fecundity and a trade-off between flight and fecundity via maternal effects.

Nuclear Factor-kappaB Enhances ErbB2-induced Mammary Tumorigenesis and Neoangiogenesis in Vivo

The American Journal of Pathology. May, 2009  |  Pubmed ID: 19349372

The (HER2/Neu) ErbB2 oncogene is commonly overexpressed in human breast cancer and is sufficient for mammary tumorigenesis in transgenic mice. Nuclear factor (NF)-kappaB activity is increased in both human and murine breast tumors. The immune response to mammary tumorigenesis may regulate tumor progression. The role of endogenous mammary epithelial cell NF-kappaB had not previously been determined in immune-competent animals. Furthermore, the role of the NF-kappaB components, p50 and p65, in tumor growth was not known. Herein, the expression of a stabilized form of the NF-kappaB-inhibiting IkappaBalpha protein (IkappaBalphaSR) in breast tumor cell lines that express oncogenic ErbB2 inhibited DNA synthesis and growth in both two- and three-dimensional cultures. Either NF-kappaB inhibition or selective silencing of p50 or p65 led to a loss of contact-independent tumor growth in vitro. IkappaBalphaSR reversed the features of the oncogene-induced phenotype under three-dimensional growth conditions. The NF-kappaB blockade inhibited ErbB2-induced mammary tumor growth in both immune-competent and immune-deficient mice. These findings were associated with both reduced tumor microvascular density and a reduction in the amount of vascular endothelial growth factor. The expression of IkappaBalphaSR in breast cancer tumors inhibited angiogenesis. Thus, mammary epithelial cell NF-kappaB activity enhances ErbB2-mediated mammary tumorigenesis in vivo by promoting both growth and survival signaling via the promotion of tumor vasculogenesis.

The Cell Fate Determination Factor Dachshund Inhibits Androgen Receptor Signaling and Prostate Cancer Cellular Growth

Cancer Research. Apr, 2009  |  Pubmed ID: 19351840

Initially isolated as the dominant suppressor of the mutant epidermal growth factor receptor (ellipse), the Dachshund gene plays a key role in metazoan development regulating the Retinal Determination Gene Network. Herein, the DACH1 gene was expressed in normal prostate epithelial cells with reduced expression in human prostate cancer. DACH1 inhibited prostate cancer cellular DNA synthesis, growth in colony forming assays, and blocked contact-independent growth in soft agar assays. DACH1 inhibited androgen receptor (AR) activity, requiring a conserved DS Domain (Dachshund domain conserved with Ski/Sno) that bound NCoR/HDAC and was recruited to an androgen-responsive gene promoter. DACH1 inhibited ligand-dependent activity of AR mutations identified in patients with androgen-insensitive prostate cancer. The DS domain was sufficient for repression of the AR wild-type but failed to repress an AR acetylation site point mutant. These studies show a role for the Retinal Determination Gene Network in regulating cellular growth and signaling in prostate cancer.

Mutation of an Aminopeptidase N Gene is Associated with Helicoverpa Armigera Resistance to Bacillus Thuringiensis Cry1Ac Toxin

Insect Biochemistry and Molecular Biology. Jul, 2009  |  Pubmed ID: 19376227

A Cry1Ac-resistant strain (Bt-R) of Helicoverpa armigera, with 2971-fold resistance, was derived by selection with Cry1Ac toxin for 75 generations. We used cDNA-amplified fragment length polymorphism analysis to identify those genes differentially expressed in the Cry1Ac-resistant and -susceptible strains, which revealed 212 differentially expressed transcripts among 2000 screened cDNAs. Among these transcript-derived fragments (TDFs), 37 showed some homology to known sequences, including Aminopeptidase N (APN), which is expressed in the midgut epithelium and has been implicated as a Cry1A subfamily receptor in several moths, including H. armigera. We confirmed the TDF by RT-PCR and identified a deletion mutation of apn1 in the Bt-R strain. We expressed the TDF in bacteria. The partial HaAPN1-96S wild-type protein, bound to Cry1Ac on ligand blots, whereas HaAPN1-BtR did not. This suggested that HaAPN1 is a receptor for Bt Cry1Ac and that its deletion mutation is associated with Cry1Ac resistance in H. armigera. The absence of one binding site is responsible for its resistance to Cry1Ac. We developed an allele-specific PCR to monitor whether the apn1 gene in an H. armigera field population produced a similar mutation. No deleted mutants were found in 2250 individuals collected from the field in 2006-2007.

Reduction of Bacillus Thuringiensis Cry1Ac Toxicity Against Helicoverpa Armigera by a Soluble Toxin-binding Cadherin Fragment

Journal of Insect Physiology. Aug, 2009  |  Pubmed ID: 19446559

A cadherin-like protein has been identified as a putative receptor for Bacillus thuringiensis (Bt) Cry1Ac toxin in Helicoverpa armigera and plays a key role in Bt insecticidal action. In this study, we produced a fragment from this H. armigera Cry1Ac toxin-binding cadherin that included the predicted toxin-binding region. Binding of Cry1Ac toxin to this cadherin fragment facilitated the formation of a 250-kDa toxin oligomer. The cadherin fragment was evaluated for its effect on Cry1Ac toxin-binding and toxicity by ligand blotting, binding assays, and bioassays. The results of ligand blotting and binding assays revealed that the binding of Cry1Ac to H. armigera midgut epithelial cells was reduced under denaturing or native conditions in vitro. Bioassay results indicated that toxicities from Cry1Ac protoxin or activated toxin were reduced in vivo by the H. armigera cadherin fragment. The addition of the cadherin fragment had no effect on Cry2Ab toxicity.

The Cell Fate Determination Factor DACH1 is Expressed in Estrogen Receptor-alpha-positive Breast Cancer and Represses Estrogen Receptor-alpha Signaling

Cancer Research. Jul, 2009  |  Pubmed ID: 19605405

The Dachshund (dac) gene, initially cloned as a dominant inhibitor of the Drosophila hyperactive EGFR mutant ellipse, encodes a key component of the cell fate determination pathway involved in Drosophila eye development. Analysis of more than 2,200 breast cancer samples showed improved survival by some 40 months in patients whose tumors expressed DACH1. Herein, DACH1 and estrogen receptor-alpha (ERalpha) expressions were inversely correlated in human breast cancer. DACH1 bound and inhibited ERalpha function. Nuclear DACH1 expression inhibited estradiol (E(2))-induced DNA synthesis and cellular proliferation. DACH1 bound ERalpha in immunoprecipitation-Western blotting, associated with ERalpha in chromatin immunoprecipitation, and inhibited ERalpha transcriptional activity, requiring a conserved DS domain. Proteomic analysis identified proline, glutamic acid, and leucine rich protein 1 (PELP1) as a DACH1-binding protein. The DACH1 COOH terminus was required for binding to PELP1. DACH1 inhibited induction of ERalpha signaling. E(2) recruited ERalpha and disengaged corepressors from DACH1 at an endogenous ER response element, allowing PELP1 to serve as an ERalpha coactivator. DACH1 expression, which is lost in poor prognosis human breast cancer, functions as an endogenous inhibitor of ERalpha function.

Cry2Ab Tolerance Response of Helicoverpa Armigera (Lepidoptera: Noctuidae) Populations from CrylAc Cotton Planting Region

Journal of Economic Entomology. Jun, 2009  |  Pubmed ID: 19610441

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is an important lepidopteran pest of cotton, Gossypium hirsutum L., in Asia. Transgenic cotton expressing the cry1Ac gene from Bacillus thuringiensis (Bt) has been widely planted for control of this pest. For managing the potential risk from resistance evolution in this pest, a new transgenic Bt cotton containing cry1Ac and cry2Ab genes (gene pyramided strategy) was designed as a successor of cry1Ac cotton. This article reports on levels of Cry2Ab tolerance in H. armigera populations from CrylAc cotton planting region in China based on bioassays of F1 and F2 offspring of isofemale lines. In total, 572 isofemale families of H. armigera from Xiajin County of Shandong Province (an intensive Bt cotton-planting area) and 124 families from Anci County of Hebei Province [a multiple-crop system, including corn (Zea mays L.), soybean (Gycine max (L.) Merr., peanut (Arachis spp.), and Bt cotton] were screened with both Cry1Ac- and Cry2Ab-containing diets in 2008. The bioassays results indicated that relative average development rates (RADR) of F1 full-sib families from field-collected female moths on Cry1Ac- and Cry2Ab-containing diet were positively correlated. The same correlation was found in the F2 generation, indicating cross-tolerance between Cry1Ac and Cry2Ab in field populations of H. armigera in Yellow River cotton-farming region of China. This cross-tolerance must be considered in evaluating the utility of pyramiding Bt genes in cotton for delaying evolution of resistance.

Frequency of Bt Resistance Alleles in H. Armigera During 2006-2008 in Northern China

Environmental Entomology. Aug, 2009  |  Pubmed ID: 19689916

Helicoverpa armigera is an important lepidopteran pest of cotton in China. From 2002, the frequency of Bt resistance alleles and quantitative shifts in larval Cry1Ac tolerance of field H. armigera population were monitored using bioassays of F(1) and F(2) offspring of isofemale lines from Xiajin County of Shandong Province (an intensive Bt cotton planting area) and Anci County of Hebei Province (a multiple-crop system including corn, soybean, peanut, and Bt cotton) in northern China. During 2006-2008, a total of 2,306 isofemale lines from the Xiajin population and a total of 1,270 isofemale lines from the Anci population were successfully screened on Cry1Ac diets. For each year, it was estimated that the major resistance gene frequency in Xiajin population in 2006, 2007, and 2008 was 0, 0.00022, and 0.00033, respectively. No major alleles conferring resistance to Cry1Ac were found in the Anci population; the frequency of resistance alleles for Cry1Ac was 0. Based on the relative average development rates (RADRs) of H. armigera larvae in F(1) tests, no substantial increase in Cry1Ac tolerance was found in either location over the 3-yr period. There were also significantly positive correlations between RADR of lines in the F(1) generation and the RADR of their F(2) offspring, indicating genetic variation in response to toxin. The low frequency of resistance alleles found in this study and in our previous results from 2002 to 2005 suggest the frequency of resistance alleles has remained low and that natural refugia resistance management strategy maybe effective for delaying resistance evolution in H. armigera to Bt cotton in northern China.

The Dachshund Gene in Development and Hormone-responsive Tumorigenesis

Trends in Endocrinology and Metabolism: TEM. Jan, 2010  |  Pubmed ID: 19896866

The dachshund (dac) gene was initially described as a mutant phenotype in flies featuring extremely short legs relative to their body length. Functioning as a dominant suppressor of the ellipse mutation, a hypermorphic allele of the Epidermal Growth Factor Receptor (EGFR), the dac gene plays a key role in metazoan development, regulating ocular, limb, brain, and gonadal development. In the Drosophila eye, dac is a key component of the Retinal Determination Gene Network (RDGN) governing the normal initiation of the morphogenetic furrow and thereby eye development. Recent studies have demonstrated an important role for human Dachshund homologue (DACH1) in tumorigenesis, in particular, breast, prostate and ovarian cancer. The molecular mechanisms by which DACH1 regulates differentiation and tumorigenesis are discussed herein.

Antisera-mediated in Vivo Reduction of Cry1Ac Toxicity in Helicoverpa Armigera

Journal of Insect Physiology. Jul, 2010  |  Pubmed ID: 20035762

A functional assessment of Bacillus thuringiensis (Bt) toxin receptors in the midgut of lepidopteran insects will facilitate understanding of the toxin mode of action and provide effective strategies to counter the development of resistance. In this study, we produced anti-aminopeptidase (APN) and anti-cadherin sera with purified Cry1Ac toxin-binding APN or cadherin fragments from Heliocoverpa armigera. Antisera were evaluated for their effects on Cry1Ac toxicity through bioassays. Our results indicated that both the anti-APN and anti-cadherin sera reduced Cry1Ac toxicity in vivo, although cadherin antiserum reduced toxicity more than APN antiserum. These results suggest that both APN and cadherin are involved in Cry1Ac intoxication of H. armigera, evidence that the pore formation model may be representative of Cry1Ac toxin mode of action in this insect.

Characterization of a Cry1Ac Toxin-binding Alkaline Phosphatase in the Midgut from Helicoverpa Armigera (Hübner) Larvae

Journal of Insect Physiology. Jun, 2010  |  Pubmed ID: 20170658

Midgut membrane-bound alkaline phosphatases (mALP) tethered to the brush border membrane surface by a glycosylphosphatidylinositol (GPI) anchor have been proposed as crucial for Cry1Ac intoxication. In the present work, two full-length cDNAs-encoding alkaline phosphatases in the midgut of Helicoverpa armigera larvae were cloned and named HaALP1 (GenBank accession no. EU729322) and HaALP2 (GenBank accession no. EU729323), respectively. These two clones displayed high identity (above 94%) at the amino acid sequence, indicating that they may represent allelic variants, and were predicted to contain a GPI anchor. Protein sequence alignment revealed that HaALPs were grouped with mALP from the Heliothis virescens midgut. The HaALP1 and HaALP2 ( approximately 68kDa) proteins were heterologously expressed in Sf9 cells using a baculovirus expression system and purified to homogeneity. Ligand blot and dot blot analysis revealed that the Cry1Ac bound to both denatured and native purified HaALPs. Data from lectin blots, competition assays with soybean agglutinin (SBA) lectin and GalNAc binding inhibition assays were indicative of the presence of GalNAc on HaALPs and binding of Cry1Ac toxin to this residue. This observation was further confirmed through N-glycosidase digestion of HaALPs, which resulted in reduced Cry1Ac binding. Our data represent the first report on HaALPs and their putative role as receptors for Cry1Ac toxin in H. armigera.

Attenuation of Forkhead Signaling by the Retinal Determination Factor DACH1

Proceedings of the National Academy of Sciences of the United States of America. Apr, 2010  |  Pubmed ID: 20351289

The Drosophila Dachshund (Dac) gene, cloned as a dominant inhibitor of the hyperactive growth factor mutant ellipse, encodes a key component of the retinal determination gene network that governs cell fate. Herein, cyclic amplification and selection of targets identified a DACH1 DNA-binding sequence that resembles the FOX (Forkhead box-containing protein) binding site. Genome-wide in silico promoter analysis of DACH1 binding sites identified gene clusters populating cellular pathways associated with the cell cycle and growth factor signaling. ChIP coupled with high-throughput sequencing mapped DACH1 binding sites to corresponding gene clusters predicted in silico and identified as weight matrix resembling the cyclic amplification and selection of targets-defined sequence. DACH1 antagonized FOXM1 target gene expression, promoter occupancy in the context of local chromatin, and contact-independent growth. Attenuation of FOX function by the cell fate determination pathway has broad implications given the diverse role of FOX proteins in cellular biology and tumorigenesis.

Electrophysiological and Behavioral Responses of Microplitis Mediator (Hymenoptera: Braconidae) to Caterpillar-induced Volatiles from Cotton

Environmental Entomology. Apr, 2010  |  Pubmed ID: 20388293

Microplitis mediator Haliday (Hymenoptera: Braconidae) is an important larval endoparasitoid of various lepidopteran pests, including Helicoverpa armigera (Hübner). In China, H. armigera is a key pest of cotton and is currently the focus of several biological control efforts that use M. mediator as principal natural enemy of this pest. To improve the success of biological control efforts, behavioral studies are needed that shed light on the interaction between M. mediator and H. armigera. In this study, we determined M. mediator response to volatile compounds from undamaged, mechanically injured, or H. armigera--damaged plants and identified attractive volatiles. In Y-tube olfactometer assays, we found that mechanically damaged plants and/or plants treated with H. armigera oral secretions did not attract wasps. However, volatiles from H. armigera-damaged plants elicited a strong attraction of both M. mediator sexes. Headspace extracts from H. armigera-damaged cotton were analyzed by coupled gas chromatography-electroantennographic detection (GC-EAD), and a total of seven different compounds were found to elicit electroantennogram (EAG) responses, including an unknown compound. Six different EAD-active volatiles were identified from caterpillar-damaged cotton plants, of which 3, 7-dimethyl-1, 3, 6-octatriene and (Z)-3-hexenyl acetate were the principal compounds. Olfactometer assays indicated that individual synthetic compounds of 3, 7-dimethyl-1, 3, 6-octatriene, (Z)-3-hexenyl acetate, and nonanal were attractive to M. mediator. Field cage studies showed that parasitism of H. armigera larvae by M. mediator was higher on cotton plants to which 3,7-dimethyl-1,3, 6-octatriene was applied. Our results show that the combination of terpenoids and green leaf volatiles may not only facilitate host, mate, or food location but may also increase H. armigera parasitism by M. mediator.

Mirid Bug Outbreaks in Multiple Crops Correlated with Wide-scale Adoption of Bt Cotton in China

Science (New York, N.Y.). May, 2010  |  Pubmed ID: 20466880

Long-term ecological effects of transgenic Bacillus thuringiensis (Bt) crops on nontarget pests have received limited attention, more so in diverse small holder-based cropping systems of the developing world. Field trials conducted over 10 years in northern China show that mirid bugs (Heteroptera: Miridae) have progressively increased population sizes and acquired pest status in cotton and multiple other crops, in association with a regional increase in Bt cotton adoption. More specifically, our analyses show that Bt cotton has become a source of mirid bugs and that their population increases are related to drops in insecticide use in this crop. Hence, alterations of pest management regimes in Bt cotton could be responsible for the appearance and subsequent spread of nontarget pests at an agro-landscape level.

Phytase Transgenic Maize Does Not Affect the Development and Nutrition Utilization of Ostrinia Furnacalis and Helicoverpa Armigera

Environmental Entomology. Jun, 2010  |  Pubmed ID: 20550822

Use of transgenic maize expressing phytase in seeds as feedstuff can greatly increase phosphate availability to livestock and poultry. Because phosphorus is an essential mineral for all living organisms, growing of phytase transgenic maize may affect the performance of the arthropod community in maize fields. We conducted a preliminary study to assess the potential effects of phytase transgenic maize (BVLA430101) on two herbivore species, Ostrinia furnacalis (Guenée) and Helicoverpa armigera (Hübner), both of which are directly exposed to high concentrations of phytase caused by ingestion of transgenic maize kernels. Our results showed that for both species, survival and duration of the first and second instars and fresh weight of the third instar were not affected when fed transgenic phytase maize kernels compared with those fed nontransformed near isoline kernels. Similarly, there was no statistical difference detected for the same life table parameters when the herbivores were fed artificial diet containing either transgenic phytase maize meal or nontransformed maize meal. In addition, the nutrition utilization of the two species was evaluated with the same diet treatments by comparing the following indices: relative food consumption rate (RCR), relative metabolic rate (RMR), efficiency of approximate digestibility (EAD), efficiency of conversation of ingested food (ECI), and efficiency of conversation of digested food (ECD). No statistical difference was detected for any index of either species between transgenic maize and nontransformed maize treatments. These results provide useful baseline information for further studies to assess the potential effects of phytase transgenic maize on other arthropods in maize fields.

Cotton Bollworm Resistance to Bt Transgenic Cotton: a Case Analysis

Science China. Life Sciences. Aug, 2010  |  Pubmed ID: 20821292

Cotton bollworm (Helicoverpa armigera) is one of the most serious insect pests of cotton. Transgenic cotton expressing Cry toxins derived from a soil bacterium, Bacillus thuringiensis (Bt), has been produced to target this pest. Bt cotton has been widely planted around the world, and this has resulted in efficient control of bollworm populations with reduced use of synthetic insecticides. However, evolution of resistance by this pest threatens the continued success of Bt cotton. To date, no field populations of bollworm have evolved significant levels of resistance; however, several laboratory-selected Cry-resistant strains of H. armigera have been obtained, which suggests that bollworm has the capacity to evolve resistance to Bt. The development of resistance to Bt is of great concern, and there is a vast body of research in this area aimed at ensuring the continued success of Bt cotton. Here, we review studies on the evolution of Bt resistance in H. armigera, focusing on the biochemical and molecular basis of Bt resistance. We also discuss resistance management strategies, and monitoring programs implemented in China, Australia, and India.

Transcription Elongation Regulator 1 is a Co-integrator of the Cell Fate Determination Factor Dachshund Homolog 1

The Journal of Biological Chemistry. Dec, 2010  |  Pubmed ID: 20956529

DACH1 (Dachshund homolog 1) is a key component of the retinal determination gene network and regulates gene expression either indirectly as a co-integrator or through direct DNA binding. The current studies were conducted to understand, at a higher level of resolution, the mechanisms governing DACH1-mediated transcriptional repression via DNA sequence-specific binding. DACH1 repressed gene transcription driven by the DACH1-responsive element (DRE). Recent genome-wide ChIP-Seq analysis demonstrated DACH1 binding sites co-localized with Forkhead protein (FOX) binding sites. Herein, DACH1 repressed, whereas FOX proteins enhanced, both DRE and FOXA-responsive element-driven gene expression. Reduced DACH1 expression using a shRNA approach enhanced FOX protein activity. As DACH1 antagonized FOX target gene expression and attenuated FOX signaling, we sought to identify limiting co-integrator proteins governing DACH1 signaling. Proteomic analysis identified transcription elongation regulator 1 (TCERG1) as the transcriptional co-regulator of DACH1 activity. The FF2 domain of TCERG1 was required for DACH1 binding, and the deletion of FF2 abolished DACH1 trans-repression function. The carboxyl terminus of DACH1 was necessary and sufficient for TCERG1 binding. Thus, DACH1 represses gene transcription through direct DNA binding to the promoter region of target genes by recruiting the transcriptional co-regulator, TCERG1.

No Refuge for Insect Pests

Nature Biotechnology. Dec, 2010  |  Pubmed ID: 21139612

Vip3Aa Tolerance Response of Helicoverpa Armigera Populations from a Cry1Ac Cotton Planting Region

Journal of Economic Entomology. Dec, 2010  |  Pubmed ID: 21309241

Transgenic cotton, Gossypium hirsutum L., that expresses the Bacillus thuringiensis (Bt) Cry1Ac toxin, holds great promise in controlling target insect pests. Evolution of resistance by target pests is the primary threat to the continued efficacy of Bt cotton. To thwart pest resistance evolution, a transgenic cotton culitvar that produces two different Bt toxins, cry1Ac and vip3A genes, was proposed as a successor of cry1Ac cotton. This article reports on levels of Vip3Aa tolerance in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) populations from the Cry1Ac cotton planting region in China based on bioassays of the F1 generation of isofemale lines. In total, 80 isofemale families of H. armigera from Xiajin county of Shandong Province (an intensive Bt cotton planting area) and 93 families from Anci county of Hebei Province (a multiple-crop system including corn [Zea mays L.] , soybean [Glycine max (L.) Merr.], peanut (Arachis hypogaea L.), and Bt cotton) were screened with a discriminating concentration of both Cry1Ac- and Vip3A-containing diets in 2009. From data on the relative average development rates and percentage of larval weight inhibition of F1 full-sib families tested simultaneously on Cry1Ac and Vip3Aa, results indicate that responses to Cry1Ac and Vip3Aa were not genetically correlated in field population ofH. armigera. This indicates that the threat of cross-resistance between Cry1Ac and Vip3A is low in field populations of H. armigera. Thus, the introduction of Vip3Aa/Cry1Ac-producing lines could delay resistance evolution in H. armigera in Bt cotton planting area of China.

Genetic Variation of Mitochondrial DNA in Chinese Populations of Pectinophora Gossypiella (Lepidoptera: Gelechiidae)

Environmental Entomology. Aug, 2010  |  Pubmed ID: 22127186

The pink bollworm Pectinophora gossypiella is an invasive pest insect that has successfully established populations in many cotton growing regions around the world. In this study, the genetic diversity and population structure of Chinese populations of P. gossypiella were evaluated using mitochondrial DNA sequence data (COII and Nad4). For comparison, individuals of Pakistan and America were also sequenced at the same two mtDNA regions. Extremely low genetic variation was observed in the two mitochondrial regions among all populations examined. Most of the populations harbored only one to two haplotypes. Although the Nad4 region showed relatively high haplotype diversity and nucleotide variation, ranging from 0.363 to 0.591 and from 0.00078 to 0.00140, respectively, there were only three haplotypes observed in this region. COII and Nad4 haplotype networks shaved one or two common haplotype(s) forming the center of a star-shaped phylogeny. Pairwise tests showed that most of the populations were not significantly differentiated from each other. The Chinese populations were differentiated from the Pakistani and American populations in the Nad4 region. The low level of population genetic variation of P. gossypiella is attributed to invasion bottlenecks, which may have been subsequently strengthened by its nonmigratory biology and the mosaic pattern of agricultural activities.

Frequency of Bt Resistance Alleles in Helicoverpa Armigera in the Xinjiang Cotton-planting Region of China

Environmental Entomology. Oct, 2010  |  Pubmed ID: 22546469

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a key insect pest of cotton in Xinjiang cotton-planting region of northwest China. In this region, cotton is grown on average ≈ 1.65 million ha (1.53 ≈ 1.80 million ha) annually in largely monoculture agricultural landscapes, similarly to cropping systems in the United States or Australia. Under such cropping regimes, naturally occurring refuges (with non-Bt crops) may be insufficient to prevent H. armigera resistance development to Bt toxins. Therefore, we assessed frequency of alleles conferring resistance to Cry1Ac toxin of F(1) and F(2) offspring of H. armigera isofemale lines from two distinct localities in the region during 2005-2009. More specifically, a total of 224 isofemale lines was collected from Korla County (≈ 70% Bt cotton adoption) and 402 lines from Shache County (≈ 5% Bt cotton planting). Subsequent offspring was screened on Cry1Ac artificial diet. From 2005 to 2009, resistance gene frequency in Korla fluctuated between 0.0000 and 0.0040, while being 0.0000-0.0008 in individuals collected from Shache, and there were no significant increases in both counties from 2005 to 2009. Relative average development rates (RADRs) of larvae in F(1) tests showed significant increases from Korla, but not in Shache. RADR of F(1) larvae is significantly correlated with RADR of F(2) offspring, indicating genetic variation in response to toxin in field H. armigera population. Although the occurrence of Cry1Ac resistance alleles was low in Xinjiang cotton-planting region of China, particular attention should be given to H. armigera resistance development in Korla County.

Cell Fate Determination Factor Dachshund Reprograms Breast Cancer Stem Cell Function

The Journal of Biological Chemistry. Jan, 2011  |  Pubmed ID: 20937839

The cell fate determination factor Dachshund was cloned as a dominant inhibitor of the hyperactive epidermal growth factor receptor ellipse. The expression of Dachshund is lost in human breast cancer associated with poor prognosis. Breast tumor-initiating cells (TIC) may contribute to tumor progression and therapy resistance. Here, endogenous DACH1 was reduced in breast cancer cell lines with high expression of TIC markers and in patient samples of the basal breast cancer phenotype. Re-expression of DACH1 reduced new tumor formation in serial transplantations in vivo, reduced mammosphere formation, and reduced the proportion of CD44(high)/CD24(low) breast tumor cells. Conversely, lentiviral shRNA to DACH1 increased the breast (B)TIC population. Genome-wide expression studies of mammary tumors demonstrated DACH1 repressed a molecular signature associated with stem cells (SOX2, Nanog, and KLF4) and genome-wide ChIP-seq analysis identified DACH1 binding to the promoter of the Nanog, KLF4, and Lin28 genes. KLF4/c-Myc and Oct4/Sox2 antagonized DACH1 repression of BTIC. Mechanistic studies demonstrated DACH1 directly repressed the Nanog and Sox2 promoters via a conserved domain. Endogenous DACH1 regulates BTIC in vitro and in vivo.

Reduced Levels of Membrane-bound Alkaline Phosphatase Are Common to Lepidopteran Strains Resistant to Cry Toxins from Bacillus Thuringiensis

PloS One. Mar, 2011  |  Pubmed ID: 21390253

Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP) as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP) were detected by two dimensional differential in-gel electrophoresis (2D-DIGE) analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR) we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests.

Construction and Analysis of CDNA Libraries from the Antennae of Male and Female Cotton Bollworms Helicoverpa Armigera (Hübner) and Expression Analysis of Putative Odorant-binding Protein Genes

Biochemical and Biophysical Research Communications. Apr, 2011  |  Pubmed ID: 21396914

Two high-quality cDNA libraries were constructed from female and male antennae of the cotton bollworm Helicoverpa armigera (Hübner). The titers were approximately 2.0 × 10⁶ pfu/ml for females and 2.3 × 10⁶ pfu/ml for males, and this complies with the test requirement. From the libraries, 1750 male ESTs and 1640 female ESTs were sequenced and further analyzed. We identified 15 olfactory genes (12 are new), and 14 of them have the characteristic six conserved cysteine residues. With the exception of OBP9, all the genes were classified as classical OBP genes. By alignment and cluster analysis, the 14 classical OBPs were divided into pheromone binding protein (PBP) genes, odorant binding protein (OBP) genes, general odorant binding protein 1 (GOBP1) genes, general odorant binding protein 2 (GOBP2) genes and antennae binding protein (ABP) genes. Among these genes, we obtained three PBP genes (PBP1-PBP3) including two new PBP genes, one new ABP gene, nine new OBP genes (OBP1-OBP9), one known GOBP1 gene and one known GOBP2 gene. Furthermore, the expression patterns of these 14 classical OBP genes were investigated in various tissues by real-time quantitative polymerase chain reaction (qPCR). The results indicated that some OBP genes are expressed differently in different sexes and tissues, but most of them are highly expressed in antennae.

Flight Mill Performance of the Lacewing Chrysoperla Sinica (Neuroptera: Chrysopidae) As a Function of Age, Temperature, and Relative Humidity

Journal of Economic Entomology. Feb, 2011  |  Pubmed ID: 21404845

The lacewing Chrysoperla sinica (Tjeder) (Neuroptera: Chrysopidae) is an important predator of several insect pests in China and has considerable potential as a biological control agent. An inoculative approach would be the releasing adults early in the season to ensure that populations are present before pest densities increase. However, an understanding of adult flight activity under different conditions is necessary to develop appropriate release strategies. Therefore, we used a 32-channel, computer-monitored flight mill system to determine the effect of age on the flight activity of unmated female and male adults. Both sexes had high total flight activity levels as well as the longest individual flight bouts 2 and 3 d after emergence. The effects of temperature (between 13 and 33 degrees C at 75% RH) and relative humidity (between 30 and 90% RH at 23 degrees C) on the flight activity of 3-d-old unmated adults also were determined. Flight activity declined at the lowest (13 degrees C) and highest (33 degrees C) temperatures tested, as well as at the lowest relative humidity (30% RH). These findings are discussed within the context of selecting the appropriate environmental conditions for releasing C. sinica.

The Type 1 Insulin-like Growth Factor Receptor and Resistance to DACH1

Cell Cycle (Georgetown, Tex.). Jun, 2011  |  Pubmed ID: 21558809

The mammalian homolog of the Drosophila dachshund gene (DACH1) has been reported as a tumor suppressor in human breast and prostate cancers. It downregulates the epidermal growth factor receptor (EGFR) and cyclin D1. The signaling pathway of the type 1 insulin-like growth factor receptor (IGF-IR) is known to be responsible for the development of resistance to treatment of human cancer with antibodies to the EGFR. We have asked whether DACH1 still exerts its tumor suppressor activity in cells dependent on the IGF-IR for growth. We find that in cells growing in IGF-1 (and unresponsive to EGF), DACH1 is devoid of tumor suppressor activity.

Sublethal Effects of Imidacloprid on Bemisia Tabaci (Hemiptera: Aleyrodidae) Under Laboratory Conditions

Journal of Economic Entomology. Jun, 2011  |  Pubmed ID: 21735901

The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most important pests in tropical and subtropical agriculture and is a key pest in greenhouse production worldwide. Current management of B. tabaci relies upon frequent applications of insecticides. Insecticide use not only directly affects pest populations through acute toxicity but also has indirect (sublethal) effects on pest physiology or behavior. In this study, we described sublethal effects of imidacloprid on adult feeding, immature development, adult fecundity, and F1 development of B. tabaci. Honeydew excretion of adults feeding on leaves treated with LC20 and LC40 concentration was significantly lower than that on untreated leaf discs. Egg production of B. tabaci adults subject to LC20 and LC40 concentrations also was less than untreated individuals. Upon transfer to untreated leaves, honeydew excretion and egg production recovered well within 24 and 48 h, respectively. Exposure to LC20 and LC40 concentrations significantly affected developmental time of B. tabaci eggs and nymphs, whereas it did not affect adult molting rate. We did not find sublethal effects on longevity and fecundity of B. tabaci adults when exposed to LC90 and LC40 concentrations for 24 h, and on egg hatching rate, nymphal mortality, and molting rate of the subsequent F1 generation. Exposure to imidacloprid at LC40 concentration significantly decreased the number of females in the F1 generation. Imidacloprid negatively affects development and reproduction of exposed individuals, and sex ratio of subsequent (F1) generation of B. tabaci, which probably disrupts B. tabaci population dynamics, slows population increase, and reduces infestation levels. Therefore, it is necessary to consider potential impact from imidacloprid for integrated management of the pest.

Seasonal Expression of Bt Proteins in Transgenic Rice Lines and the Resistance Against Asiatic Rice Borer Chilo Suppressalis (Walker)

Environmental Entomology. Oct, 2011  |  Pubmed ID: 22251743

Laboratory bioassays and field surveys were carried out to compare the resistance of three transgenic rice (Oryza sativa L.) lines including Bt-DL expressing a single gene cry1Ab, Bt-KF6 expressing stacked genes cry1Ac and CpTI genes and Bt-SY63 expressing a fusion gene cry1Ab/cry1Ac, respectively, to an important rice pest Chilo suppressalis (Walker). In addition, enzyme-linked immunosorbent assays (ELISA) were conducted to monitor the Bt protein expressions in rice leaves and stems at different rice growth stages. Results showed that all the transgenic rice lines exhibited significantly high resistance to the pest compared with their corresponding nontransformed isolines. Among the transgenic rice lines, Bt-SY63 and Bt-KF6 had higher resistance to C. suppressalis at early growth stage, but lower resistance at late stages, while the pest resistance of Bt-DL was relatively stable throughout the growing season. The results were consistent with ELISA results showing that Bt protein levels in Bt-SY63 or Bt-KF6 leaves decreased in late growth stages, but were relatively stable in Bt-DL at all growth stages. This demonstrates that the resistance to a pest by Bt plants is positively correlated with Cry protein expression levels in plant tissues. Compared with Bt-SY63 and Bt-KF6, the Bt protein expression levels were significantly lower in Bt-DL, while its resistance to C. suppressalis was the highest. This may suggest that C. suppressalis is more susceptible to Cry1Ab than to Cry1Ac. The data from the current study are valuable for decision-making for commercial use of Bt rice lines and development of appropriate pest control and resistance management strategies for the transgenic rice lines.

Increased Frequency of Pink Bollworm Resistance to Bt Toxin Cry1Ac in China

PloS One. 2012  |  Pubmed ID: 22238687

Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) kill some key insect pests, but evolution of resistance by pests can reduce their efficacy. The main approach for delaying pest adaptation to Bt crops uses non-Bt host plants as "refuges" to increase survival of susceptible pests. To delay evolution of pest resistance to transgenic cotton producing Bt toxin Cry1Ac, the United States and some other countries have required refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. The "natural" refuge strategy focuses on cotton bollworm (Helicoverpa armigera), the primary target of Bt cotton in China that attacks many crops, but it does not apply to another major pest, pink bollworm (Pectinophora gossypiella), which feeds almost entirely on cotton in China. Here we report data showing field-evolved resistance to Cry1Ac by pink bollworm in the Yangtze River Valley of China. Laboratory bioassay data from 51 field-derived strains show that the susceptibility to Cry1Ac was significantly lower during 2008 to 2010 than 2005 to 2007. The percentage of field populations yielding one or more survivors at a diagnostic concentration of Cry1Ac increased from 0% in 2005-2007 to 56% in 2008-2010. However, the median survival at the diagnostic concentration was only 1.6% from 2008 to 2010 and failure of Bt cotton to control pink bollworm has not been reported in China. The early detection of resistance reported here may promote proactive countermeasures, such as a switch to transgenic cotton producing toxins distinct from Cry1A toxins, increased planting of non-Bt cotton, and integration of other management tactics together with Bt cotton.

Lethal Effect of Imidacloprid on the Coccinellid Predator Serangium Japonicum and Sublethal Effects on Predator Voracity and on Functional Response to the Whitefly Bemisia Tabaci

Ecotoxicology (London, England). Jul, 2012  |  Pubmed ID: 22447470

Neonicotinoid insecticides are widely used for controlling sucking pests, and sublethal effects can be expected in beneficial arthropods like natural enemies. Serangium japonicum is an important predator in many agricultural systems in China, and a potential biological control agent against Bemisia tabaci. We evaluated the toxicity of imidacloprid to S. japonicum and its impact on the functional response to B. tabaci eggs. S. japonicum adults exposed through contact to dried residues of imidacloprid at the recommended field rate on cotton against B. tabaci (4 g active ingredient per 100 l, i.e. 40 ppm [part per million]), and reduced rates (25, 20, 15 and 10 ppm) for 24 h showed high mortality rates. The mortality induced by a lowest rate, 5 ppm, was not significantly different than the control group and thus it was considered as a sublethal rate. The lethal rate 50 and hazard quotient (HQ) were estimated to be 11.54 ppm and 3.47 respectively, indicating a risk for S. japonicum in treated fields (HQ > 2). When exposed to dried residues of imidacloprid at the sublethal rate (5 ppm) on cotton leaves, functional response of S. japonicum to B. tabaci eggs was affected with an increase in handling time and a reduction in peak consumption of eggs. Imidacloprid residues also disturbed predator voracity, the number of B. tabaci eggs consumed on treated leaves being significantly lower than on untreated leaves. All effects disappeared within a few hours after transfer to untreated cotton leaves. Imidacloprid systemically applied at the recommended field rate (for cotton) showed no toxicity to S. japonicum, nor affected the functional response of the predator. Sublethal effects of imidacloprid on S. japonicum observed in our study likely negatively affect S. japonicum development and reproductive capacity and may ultimately reduce predator population growth. These results hint at the importance of assessing potential effects of imidacloprid on S. japonicum for developing effective integrated pest management programs of B. tabaci in China.

Early Detection of Field-evolved Resistance to Bt Cotton in China: Cotton Bollworm and Pink Bollworm

Journal of Invertebrate Pathology. Jul, 2012  |  Pubmed ID: 22537835

Transgenic crops producing Bacillus thuringiensis (Bt) toxins kill some major insect pests, but pests can evolve resistance and thereby reduce the effectiveness of such Bt crops. The main approach for slowing pest adaptation to Bt crops uses non-Bt host plants as "refuges" to increase survival of susceptible pests. To delay evolution of pest resistance to cotton producing Bt toxin Cry1Ac, several countries have required refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. This strategy is designed for cotton bollworm (Helicoverpa armigera), which attacks many crops and is the primary target of Bt cotton in China, but it does not apply to pink bollworm (Pectinophora gossypiella), which feeds almost entirely on cotton in China. Here we review evidence of field-evolved resistance to Cry1Ac by cotton bollworm in northern China and by pink bollworm in the Yangtze River Valley of China. For both pests, results of laboratory diet bioassays reveal significantly decreased susceptibility of field populations to Cry1Ac, yet field control failures of Bt cotton have not been reported. The early detection of resistance summarized here may spur countermeasures such as planting Bt cotton that produces two or more distinct toxins, increased planting of non-Bt cotton, and integration of other management tactics together with Bt cotton.

Diverse Genetic Basis of Field-evolved Resistance to Bt Cotton in Cotton Bollworm from China

Proceedings of the National Academy of Sciences of the United States of America. Jun, 2012  |  Pubmed ID: 22689968

Evolution of pest resistance reduces the efficacy of insecticidal proteins from Bacillus thuringiensis (Bt) used in sprays or in transgenic crops. Although several pests have evolved resistance to Bt crops in the field, information about the genetic basis of field-evolved resistance to Bt crops has been limited. In particular, laboratory-selected resistance to Bt toxin Cry1Ac based on recessive mutations in a gene encoding a toxin-binding cadherin protein has been identified in three major cotton pests, but previous work has not determined if such mutations are associated with field-selected resistance to Bt cotton. Here we show that the most common resistance alleles in field populations of cotton bollworm, Helicoverpa armigera, selected with Bt cotton in northern China, had recessive cadherin mutations, including the deletion mutation identified via laboratory selection. However, unlike all previously studied cadherin resistance alleles, one field-selected cadherin resistance allele conferred nonrecessive resistance. We also detected nonrecessive resistance that was not genetically linked with the cadherin locus. In field-selected populations, recessive cadherin alleles accounted for 75-84% of resistance alleles detected. However, most resistance alleles occurred in heterozygotes and 59-94% of resistant individuals carried at least one nonrecessive resistance allele. The results suggest that resistance management strategies must account for diverse resistance alleles in field-selected populations, including nonrecessive alleles.

Widespread Adoption of Bt Cotton and Insecticide Decrease Promotes Biocontrol Services

Nature. Jul, 2012  |  Pubmed ID: 22722864

Over the past 16 years, vast plantings of transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have helped to control several major insect pests and reduce the need for insecticide sprays. Because broad-spectrum insecticides kill arthropod natural enemies that provide biological control of pests, the decrease in use of insecticide sprays associated with Bt crops could enhance biocontrol services. However, this hypothesis has not been tested in terms of long-term landscape-level impacts. On the basis of data from 1990 to 2010 at 36 sites in six provinces of northern China, we show here a marked increase in abundance of three types of generalist arthropod predators (ladybirds, lacewings and spiders) and a decreased abundance of aphid pests associated with widespread adoption of Bt cotton and reduced insecticide sprays in this crop. We also found evidence that the predators might provide additional biocontrol services spilling over from Bt cotton fields onto neighbouring crops (maize, peanut and soybean). Our work extends results from general studies evaluating ecological effects of Bt crops by demonstrating that such crops can promote biocontrol services in agricultural landscapes.

The Halo Effect: Suppression of Pink Bollworm on Non-Bt Cotton by Bt Cotton in China

PloS One. 2012  |  Pubmed ID: 22848685

In some previously reported cases, transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have suppressed insect pests not only in fields planted with such crops, but also regionally on host plants that do not produce Bt toxins. Here we used 16 years of field data to determine if Bt cotton caused this "halo effect" against pink bollworm (Pectinophora gossypiella) in six provinces of the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We found that Bt cotton significantly decreased the population density of pink bollworm on non-Bt cotton, with net decreases of 91% for eggs and 95% for larvae on non-Bt cotton after 11 years of Bt cotton use. Insecticide sprays targeting pink bollworm and cotton bollworm (Helicoverpa armigera) decreased by 69%. Previously reported evidence of the early stages of evolution of pink bollworm resistance to Bt cotton in China has raised concerns that if unchecked, such resistance could eventually diminish or eliminate the benefits of Bt cotton. The results reported here suggest that it might be possible to find a percentage of Bt cotton lower than the current level that causes sufficient regional pest suppression and reduces the risk of resistance.

Complete Genome Sequence of a Monosense Densovirus Infecting the Cotton Bollworm, Helicoverpa Armigera

Journal of Virology. Oct, 2012  |  Pubmed ID: 22966197

Densoviruses (DNVs) infecting arthropods are members of the family Parvoviridae. Here we report the complete genome sequence of a novel DNV with a monosense genome that infects cotton bollworms (Helicoverpa armigera), named HaDNV-1. Alignment and phylogenetic analysis revealed that HaDNV-1 showed high identity with the genus Iteravirus.

Early Season Host Plants of Apolygus Lucorum (Heteroptera: Miridae) in Northern China

Journal of Economic Entomology. Oct, 2012  |  Pubmed ID: 23156156

Apolygus lucorum (Meyer-Dür) (Heteroptera: Miridae) has become a severe pest of cotton and many other crops in northern China as a result of the widespread adoption of Bacillus thuringiensis (Berliner) cotton, with a corresponding reduction of broad-spectrum insecticide application in cotton fields. From the middle of April to middle June, A. lucorum feeds and develops on other host plants before dispersing to cotton fields. Effective suppression of A. lucorum populations before they enter cotton fields may be an excellent strategy for reducing the occurrence and damage of their subsequent generations in cotton fields. For that, basic information about the host plant range of A. lucorum during the early season is needed. Between 2006 and 2010, a total of 94 plant species from 41 families covering 39,956 square meters of land in natural conditions were sampled using the standard white pan beat method. Sixty-six plant species, including 45 weeds, 10 fruit trees, 5 timber trees, 4 pasture crops, and 2 arable crops were found to be hosts of A. lucorum. Among these species, Descurainia sophia (L.) Webb ex Prantl, Humulus scandens (Loureiro) Merrill, Zizyphus jujuba Miller, Vitis vinifera L., Viciafaba L., and Medicago sativa L. were identified as dominant host species because of their wide distribution and high population densities of A. lucorum. The results of this study provide useful information about the early season host range of A. lucorum, which can be used to develop effective strategies to control the pest before its dispersal to cotton fields.

Double Homozygous Missense Mutations in DACH1 and BMP4 in a Patient with Bilateral Cystic Renal Dysplasia

Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association. Jan, 2013  |  Pubmed ID: 23262432

Renal hypodysplasia (RHD) is characterized by small and/or disorganized kidneys following abnormal organogenesis. Mutations in several genes have been identified recently to be associated with RHD in humans, including BMP4, a member of the transforming growth factor (TGF)-β family of growth factors. DACH1 has been proposed as a candidate gene for RHD because of its involvement in the EYA-SIX-DACH network of renal developmental genes. Here, we present a patient with renal dysplasia carrying homozygous missense mutations in both BMP4 (p.N150K) and DACH1 (p.R684C). The genotype-phenotype correlation in the family hints at an oligogenic mode of inheritance of the disease in this kindred. Functional analyses of the identified DACH1 mutation in HEK293T cells demonstrated enhanced suppression of the TGF-β pathway suggesting that both mutations could act synergistically in the development of the phenotype in this patient. This finding provides a model for RHD as an oligo-/polygenic disorder and supports a role for DACH1 in the development of RHD in humans.

Dachshund Binds P53 to Block the Growth of Lung Adenocarcinoma Cells

Cancer Research. Jun, 2013  |  Pubmed ID: 23492369

Hyperactive EGF receptor (EGFR) and mutant p53 are common genetic abnormalities driving the progression of non-small cell lung cancer (NSCLC), the leading cause of cancer deaths in the world. The Drosophila gene Dachshund (Dac) was originally cloned as an inhibitor of hyperactive EGFR alleles. Given the importance of EGFR signaling in lung cancer etiology, we examined the role of DACH1 expression in lung cancer development. DACH1 protein and mRNA expression was reduced in human NSCLC. Reexpression of DACH1 reduced NSCLC colony formation and tumor growth in vivo via p53. Endogenous DACH1 colocalized with p53 in a nuclear, extranucleolar location, and shared occupancy of -15% of p53-bound genes in ChIP sequencing. The C-terminus of DACH1 was necessary for direct p53 binding, contributing to the inhibition of colony formation and cell-cycle arrest. Expression of the stem cell factor SOX2 was repressed by DACH1, and SOX2 expression was inversely correlated with DACH1 in NSCLC. We conclude that DACH1 binds p53 to inhibit NSCLC cellular growth.

Assessment of Potential Sublethal Effects of Various Insecticides on Key Biological Traits of the Tobacco Whitefly, Bemisia Tabaci

International Journal of Biological Sciences. 2013  |  Pubmed ID: 23494876

The tobacco whitefly Bemisia tabaci is one of the most devastating pests worldwide. Current management of B. tabaci relies upon the frequent applications of insecticides. In addition to direct mortality by typical acute toxicity (lethal effect), insecticides may also impair various key biological traits of the exposed insects through physiological and behavioral sublethal effects. Identifying and characterizing such effects could be crucial for understanding the global effects of insecticides on the pest and therefore for optimizing its management in the crops. We assessed the effects of sublethal and low-lethal concentrations of four widely used insecticides on the fecundity, honeydew excretion and feeding behavior of B. tabaci adults. The probing activity of the whiteflies feeding on treated cotton seedlings was recorded by an Electrical Penetration Graph (EPG). The results showed that imidacloprid and bifenthrin caused a reduction in phloem feeding even at sublethal concentrations. In addition, the honeydew excretions and fecundity levels of adults feeding on leaf discs treated with these concentrations were significantly lower than the untreated ones. While, sublethal concentrations of chlorpyrifos and carbosulfan did not affect feeding behavior, honeydew excretion and fecundity of the whitefly. We demonstrated an antifeedant effect of the imidacloprid and bifenthrin on B. tabaci, whereas behavioral changes in adults feeding on leaves treated with chlorpyrifos and carbosulfan were more likely caused by the direct effects of the insecticides on the insects' nervous system itself. Our results show that aside from the lethal effect, the sublethal concentration of imidacloprid and bifenthrin impairs the phloem feeding, i.e. the most important feeding trait in a plant protection perspective. Indeed, this antifeedant property would give these insecticides potential to control insect pests indirectly. Therefore, the behavioral effects of sublethal concentrations of imidacloprid and bifenthrin may play an important role in the control of whitefly pests by increasing the toxicity persistence in treated crops.

Expression of Cry1Ac in Transgenic Bt Soybean Lines and Their Efficiency in Controlling Lepidopteran Pests

Pest Management Science. Dec, 2013  |  Pubmed ID: 23564718

Two transgenic lines of the soybean Glycine max, MON87701 expressing the Cry1Ac protein and MON87701RR2Y expressing Cry1Ac +  EPSPS proteins, were evaluated for their resistance to four lepidopteran pests in the laboratory using detached-leaf bioassays throughout the soybean growth seasons (before anthesis, during anthesis and after anthesis) in China. Enzyme-linked immunosorbent assays (ELISAs) were used to monitor the Cry1Ac expression in soybean leaves.

The Endosymbiont Arsenophonus is Widespread in Soybean Aphid, Aphis Glycines, but Does Not Provide Protection from Parasitoids or a Fungal Pathogen

PloS One. 2013  |  Pubmed ID: 23614027

Aphids commonly harbor bacterial facultative symbionts that have a variety of effects upon their aphid hosts, including defense against hymenopteran parasitoids and fungal pathogens. The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is infected with the symbiont Arsenophonus sp., which has an unknown role in its aphid host. Our research goals were to document the infection frequency and diversity of the symbiont in field-collected soybean aphids, and to determine whether Arsenophonus is defending soybean aphid against natural enemies. We performed diagnostic PCR and sequenced four Arsenophonus genes in soybean aphids from their native and introduced range to estimate infection frequency and genetic diversity, and found that Arsenophonus infection is highly prevalent and genetically uniform. To evaluate the defensive role of Arsenophonus, we cured two aphid genotypes of their natural Arsenophonus infection through ampicillin microinjection, resulting in infected and uninfected isolines within the same genetic background. These isolines were subjected to parasitoid assays using a recently introduced biological control agent, Binodoxys communis [Braconidae], a naturally recruited parasitoid, Aphelinus certus [Aphelinidae], and a commercially available biological control agent, Aphidius colemani [Braconidae]. We also assayed the effect of the common aphid fungal pathogen, Pandora neoaphidis (Remaudiere & Hennebert) Humber (Entomophthorales: Entomophthoraceae), on the same aphid isolines. We did not find differences in successful parasitism for any of the parasitoid species, nor did we find differences in P. neoaphidis infection between our treatments. Our conclusion is that Arsenophonus does not defend its soybean aphid host against these major parasitoid and fungal natural enemies.

EYA1 Phosphatase Function is Essential to Drive Breast Cancer Cell Proliferation Through Cyclin D1

Cancer Research. Jul, 2013  |  Pubmed ID: 23636126

The Drosophila Eyes Absent Homologue 1 (EYA1) is a component of the retinal determination gene network and serves as an H2AX phosphatase. The cyclin D1 gene encodes the regulatory subunits of a holoenzyme that phosphorylates and inactivates the pRb protein. Herein, comparison with normal breast showed that EYA1 is overexpressed with cyclin D1 in luminal B breast cancer subtype. EYA1 enhanced breast tumor growth in mice in vivo, requiring the phosphatase domain. EYA1 enhanced cellular proliferation, inhibited apoptosis, and induced contact-independent growth and cyclin D1 abundance. The induction of cellular proliferation and cyclin D1 abundance, but not apoptosis, was dependent upon the EYA1 phosphatase domain. The EYA1-mediated transcriptional induction of cyclin D1 occurred via the AP-1-binding site at -953 and required the EYA1 phosphatase function. The AP-1 mutation did not affect SIX1-dependent activation of cyclin D1. EYA1 was recruited in the context of local chromatin to the cyclin D1 AP-1 site. The EYA1 phosphatase function determined the recruitment of CBP, RNA polymerase II, and acetylation of H3K9 at the cyclin D1 gene AP-1 site regulatory region in the context of local chromatin. The EYA1 phosphatase regulates cell-cycle control via transcriptional complex formation at the cyclin D1 promoter.

Epigenetic Silencing of DACH1 Induces Loss of Transforming Growth Factor-β1 Antiproliferative Response in Human Hepatocellular Carcinoma

Hepatology (Baltimore, Md.). Dec, 2013  |  Pubmed ID: 23787902

Human dachshund homolog 1 (DACH1) is a major component of the Retinal Determination Gene Network (RDGN) and functions as a tumor suppressor. However, the regulation of DACH1 expression and its function in hepatocellular carcinoma (HCC) remain unclear. In this study, epigenetic changes of DACH1 were analyzed in HCC cell lines and primary cancers. We found that promoter region hypermethylation was correlated with loss or reduction of DACH1 expression, and restoration of DACH1 expression was induced by 5-aza-2'-deoxycytidine (5-AZA) in HCC cell lines. Promoter region methylation was found in 42% of primary HCC. Reduced expression of DACH1 was associated with poor differentiation of HCC nodules and higher serum aspartate aminotransferase/alanine aminotransferase ratio. DACH1 suppressed cellular growth by reactivating transforming growth factor beta (TGF-β) signaling. Ectopic expression of DACH1 enhanced chemosensitivity to 5-fluorouracil (5-FU) by inducing p21 expression in HCC cells.

Acetylation of the Cell-fate Factor Dachshund Determines P53 Binding and Signaling Modules in Breast Cancer

Oncotarget. Jun, 2013  |  Pubmed ID: 23798621

Breast cancer is a leading form of cancer in the world. The Drosophila Dac gene was cloned as an inhibitor of the hyperactive epidermal growth factor (EGFR), ellipse. Herein, endogenous DACH1 co-localized with p53 in a nuclear, extranucleolar compartment and bound to p53 in human breast cancer cell lines, p53 and DACH1 bound common genes in Chip-Seq. Full inhibition of breast cancer contact-independent growth by DACH1 required p53. The p53 breast cancer mutants R248Q and R273H, evaded DACH1 binding. DACH1 phosphorylation at serine residue (S439) inhibited p53 binding and phosphorylation at p53 amino-terminal sites (S15, S20) enhanced DACH1 binding. DACH1 binding to p53 was inhibited by NAD-dependent deacetylation via DACH1 K628. DACH1 repressed p21CIP1 and induced RAD51, an association found in basal breast cancer. DACH1 inhibits breast cancer cellular growth in an NAD and p53-dependent manner through direct protein-protein association.

Combination of Plant and Insect Eggs As Food Sources Facilitates Ovarian Development in an Omnivorous Bug Apolygus Lucorum (Hemiptera: Miridae)

Journal of Economic Entomology. Jun, 2013  |  Pubmed ID: 23865184

Diet nutrient is considered as an important regulatory factor for reproduction of insects. To understand the effect of different food sources on the reproductive physiology of Apolygus lucorum (Meyer-Dür), the ovarian development in adult females was investigated when they were fed on green beans (Gb), combination of green beans Phaseolus vulgaris L and Helicoverpa armigera eggs (GbHe), or H. armigera eggs (He). A female of A. lucorum has two ovaries, and each ovary contained seven yellowish ovarioles. Females fed on Gb or GbHe had larger ovaries and the ovarioles contained larger numbers of oocytes compared with those fed on He. Females in GeHe treatment has significantly higher number of follicles per ovary throughout the whole adult period compared with those in Gb or He treatment. Furthermore, the length of the best developed ovariole was affected by the diet type. The females fed on GbHe had the most developed ovarioles, with significantly longer ovarioles than those fed on Gb or He. A method was described to quantitatively score the degree of ovarian development in the current study. Similarly, the ovarian development scores were significantly higher for females in GbHe treatment than those in other two diet treatments. The ovarian development significantly delayed for females fed on He. Our results demonstrate that A. lucorum, as an omnivorous insect species, can acquire nutrients from both plant and animal origin food sources, and the combination of plants and animal food sources can significantly facilitate the ovary development of its females.

Preference of a Polyphagous Mirid Bug, Apolygus Lucorum (Meyer-Dür) for Flowering Host Plants

PloS One. 2013  |  Pubmed ID: 23874835

Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is one of the most important herbivores in a broad range of cultivated plants, including cotton, cereals, vegetables, and fruit crops in China. In this manuscript, we report on a 6-year long study in which (adult) A. lucorum abundance was recorded on 174 plant species from 39 families from early July to mid-September. Through the study period per year, the proportion of flowering plants exploited by adult A. lucorum was significantly greater than that of non-flowering plants. For a given plant species, A. lucorum adults reached peak abundance at the flowering stage, when the plant had the greatest attraction to the adults. More specifically, mean adult abundance on 26 species of major host plants and their relative standard attraction were 10.3-28.9 times and 9.3-19.5 times higher at flowering stage than during non-flowering periods, respectively. Among all the tested species, A. lucorum adults switched food plants according to the succession of flowering plant species. In early July, A. lucorum adults preferred some plant species in bloom, such as Vigna radiata, Gossypium hirsutum, Helianthus annuus and Chrysanthemum coronarium; since late July, adults dispersed into other flowering hosts (e.g. Ricinus communis, Impatiens balsamina, Humulus scandens, Ocimum basilicum, Agastache rugosus and Coriandrum sativum); in early September, they largely migrated to flowering Artemisia spp. (e.g. A. argyi, A. lavandulaefolia, A. annua and A. scoparia). Our findings underscore the important role of flowering plays in the population dynamics and inter-plant migration of this mirid bug. Also, our work helps understand evolutionary aspects of host plant use in polyphagous insects such as A. lucorum, and provides baseline information for the development of sustainable management strategies of this key agricultural pest.

Epigenetic Regulation of DACH1, a Novel Wnt Signaling Component in Colorectal Cancer

Epigenetics. Dec, 2013  |  Pubmed ID: 24149323

Colorectal cancer (CRC) is one of the common malignant tumors worldwide. Both genetic and epigenetic changes are regarded as important factors of colorectal carcinogenesis. Loss of DACH1 expression was found in breast, prostate, and endometrial cancer. To analyze the regulation and function of DACH1 in CRC, 5 colorectal cancer cell lines, 8 cases of normal mucosa, 15 cases of polyps and 100 cases of primary CRC were employed in this study. In CRC cell lines, loss of DACH1 expression was correlated with promoter region hypermethylation, and re-expression of DACH1 was induced by 5-Aza-2'-deoxyazacytidine treatment. We found that DACH1 was frequently methylated in primary CRC and this methylation was associated with reduction in DACH1 expression. These results suggest that DACH1 expression is regulated by promoter region hypermethylation in CRC. DACH1 methylation was associated with late tumor stage, poor differentiation, and lymph node metastasis. Re-expression of DACH1 reduced TCF/LEF luciferase reporter activity and inhibited the expression of Wnt signaling downstream targets (c-Myc and cyclinD1). In xenografts of HCT116 cells in which DACH1 was re-expressed, tumor size was smaller than in controls. In addition, restoration of DACH1 expression induced G2/M phase arrest and sensitized HCT116 cells to docetaxel. DACH1 suppresses CRC growth by inhibiting Wnt signaling both in vitro and in vivo. Silencing of DACH1 expression caused resistance of CRC cells to docetaxel. In conclusion, DACH1 is frequently methylated in human CRC and methylation of DACH1 may serve as detective and prognostic marker in CRC.

The Evolution and Expression of the Moth Visual Opsin Family

PloS One. 2013  |  Pubmed ID: 24205129

Because visual genes likely evolved in response to their ambient photic environment, the dichotomy between closely related nocturnal moths and diurnal butterflies forms an ideal basis for investigating their evolution. To investigate whether the visual genes of moths are associated with nocturnal dim-light environments or not, we cloned long-wavelength (R), blue (B) and ultraviolet (UV) opsin genes from 12 species of wild-captured moths and examined their evolutionary functions. Strong purifying selection appeared to constrain the functions of the genes. Dark-treatment altered the levels of mRNA expression in Helicoverpa armigera such that R and UV opsins were up-regulated after dark-treatment, the latter faster than the former. In contrast, B opsins were not significantly up-regulated. Diel changes of opsin mRNA levels in both wild-captured and lab-reared individuals showed no significant fluctuation within the same group. However, the former group had significantly elevated levels of expression compared with the latter. Consequently, environmental conditions appeared to affect the patterns of expression. These findings and the proportional expression of opsins suggested that moths potentially possessed color vision and the visual system played a more important role in the ecology of moths than previously appreciated. This aspect did not differ much from that of diurnal butterflies.

Cyclin D1 Induction of Dicer Governs MicroRNA Processing and Expression in Breast Cancer

Nature Communications. 2013  |  Pubmed ID: 24287487

Cyclin D1 encodes the regulatory subunit of a holoenzyme that phosphorylates the pRB protein and promotes G1/S cell-cycle progression and oncogenesis. Dicer is a central regulator of miRNA maturation, encoding an enzyme that cleaves double-stranded RNA or stem-loop-stem RNA into 20-25 nucleotide long small RNA, governing sequence-specific gene silencing and heterochromatin methylation. The mechanism by which the cell cycle directly controls the non-coding genome is poorly understood. Here we show that cyclin D1(-/-) cells are defective in pre-miRNA processing which is restored by cyclin D1a rescue. Cyclin D1 induces Dicer expression in vitro and in vivo. Dicer is transcriptionally targeted by cyclin D1, via a cdk-independent mechanism. Cyclin D1 and Dicer expression significantly correlates in luminal A and basal-like subtypes of human breast cancer. Cyclin D1 and Dicer maintain heterochromatic histone modification (Tri-m-H3K9). Cyclin D1-mediated cellular proliferation and migration is Dicer-dependent. We conclude that cyclin D1 induction of Dicer coordinates microRNA biogenesis.

Involvement of Nonbinding Site Proteinases in the Development of Resistance of Helicoverpa Armigera (Lepidoptera: Noctuidae) to Cry1Ac

Journal of Economic Entomology. Dec, 2013  |  Pubmed ID: 24498753

Development of resistance to transgenic crops expressing the Cry toxin from Bacterium thuringiensis (Bt) has been the major concern for the long-term success of Bt crops. Alterations in nonbinding site proteinases and Bt toxin receptors are the two types of mechanisms responsible for Bt resistance in resistant insects. However, little is known about the relative contributions of the two types of mechanisms in the early and late phases of the development of Bt resistance. To address the relative contributions of four nonbinding site proteinases including esterase, total protease, chymotrypsin, and glutathione S-transferase in the early and late phases of the development of Cry1Ac resistance, we analyzed the relationships between nonbinding site proteinases and resistance of three groups of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) strains with different resistance levels because of different geographic origins and selection pressures. Positive correlation (esterase, glutathione-S-tranferases [GST], and chymotrypsin) and negative correlation (total midgut protease) were observed within the low to moderate group II resistant strains. Such correlations were less obvious within the low to moderate group III resistant strains because of only threefold differences in LC50 values. Relative to the unselected susceptible 96S strain, the two highly resistant group I resistant strains BtI and BtR have the same amounts of esterase, GST, and chymotrypsin and disproportionally decreased the amount of total midgut protease. Overall, the low to moderate resistant strains had the lowest amount of the nonbinding site proteinases. The results obtained suggest that alternations in the nonbinding site proteinases probably can only confer low to moderate levels of resistance and thus are enriched in the early phase of the development of Cry1Ac resistance.

Cell Fate Factor DACH1 Represses YB-1-mediated Oncogenic Transcription and Translation

Cancer Research. Feb, 2014  |  Pubmed ID: 24335958

The epithelial-mesenchymal transition (EMT) enhances cellular invasiveness and confers tumor cells with cancer stem cell-like characteristics, through transcriptional and translational mechanisms. The mechanisms maintaining transcriptional and translational repression of EMT and cellular invasion are poorly understood. Herein, the cell fate determination factor Dachshund (DACH1), suppressed EMT via repression of cytoplasmic translational induction of Snail by inactivating the Y box-binding protein (YB-1). In the nucleus, DACH1 antagonized YB-1-mediated oncogenic transcriptional modules governing cell invasion. DACH1 blocked YB-1-induced mammary tumor growth and EMT in mice. In basal-like breast cancer, the reduced expression of DACH1 and increased YB-1 correlated with poor metastasis-free survival. The loss of DACH1 suppression of both cytoplasmic translational and nuclear transcriptional events governing EMT and tumor invasion may contribute to poor prognosis in basal-like forms of breast cancer, a relatively aggressive disease subtype.

Use of a Pollen-based Diet to Expose the Ladybird Beetle Propylea Japonica to Insecticidal Proteins

PloS One. 2014  |  Pubmed ID: 24409328

A rape seed pollen-based diet was developed and found to be suitable for use in a dietary exposure assay for Propylea japonica. Using the diet, we established and validated a dietary exposure assay by using the protease inhibitor E-64 as positive control. Dose-dependent responses were documented for all observed life-table parameters of P. japonica including survival, pupation and eclosion rates, development time and adult weight. Results suggested that the dietary assay can detect the effects of insecticidal compounds on the survival and development of P. japonica. Using the established dietary assay, we subsequently tested the toxicity of Cry1Ab, Cry1Ac and Cry1F proteins that are expressed by transgenic maize, cotton or rice plants to P. japonica larvae. The diet containing E-64 was included as a positive control. Survival and development of P. japonica larvae were not adversely affected when the diet contained purified Cry1Ab, Cry1Ac, or Cry1F at 500 µg/g diet representing a worst-case exposure scenario. In contrast, P. japonica larvae were adversely affected when the diet contained E-64. The bioactivity and stability of the Cry proteins in the diet and Cry protein uptake by the ladybird larvae were confirmed by bioassay with a Cry-sensitive insect species and by ELISA. The current study describes a suitable experimental system for assessing the potential effects of gut-active insecticidal compounds on ladybird beetle larvae. The experiments with the Cry proteins demonstrate that P. japonica larvae are not sensitive to Cry1Ab, Cry1Ac and Cry1F.

Effect of Farm Management Practices in the Bt Toxin Production by Bt Cotton: Evidence from Farm Fields in China

Transgenic Research. Jun, 2014  |  Pubmed ID: 24445856

Based on farm field plot level survey data and laboratory test, we examine the determinants of the expression of Bt toxin in China's Bt cotton production. The results show that the expression of Bt toxin differs significantly among varieties. Even for the same variety the expression of Bt toxin also varies substantially among villages and among farmers in the same village. Econometric analyses show that after controlling for the effects of varieties and locations (or villages), farm management, particular applications of phosphate and potash fertilizers, and manure, has significant positive effects on Bt toxin expression in farmer's fields. In contrast to previous studies which showed that nitrogen fertilizer has a positive impact on expression of Bt toxin, this study shows that nitrogen fertilizer has no significant impact on expression of Bt toxin in farmer's fields. On the other hand, the expression of Bt toxin has a positive relationship with phosphate fertilizer, potash fertilizer and manure application.

Biosafety Management and Commercial Use of Genetically Modified Crops in China

Plant Cell Reports. Apr, 2014  |  Pubmed ID: 24493253

As a developing country with relatively limited arable land, China is making great efforts for development and use of genetically modified (GM) crops to boost agricultural productivity. Many GM crop varieties have been developed in China in recent years; in particular, China is playing a leading role in development of insect-resistant GM rice lines. To ensure the safe use of GM crops, biosafety risk assessments are required as an important part of the regulatory oversight of such products. With over 20 years of nationwide promotion of agricultural biotechnology, a relatively well-developed regulatory system for risk assessment and management of GM plants has been developed that establishes a firm basis for safe use of GM crops. So far, a total of seven GM crops involving ten events have been approved for commercial planting, and 5 GM crops with a total of 37 events have been approved for import as processing material in China. However, currently only insect-resistant Bt cotton and disease-resistant papaya have been commercially planted on a large scale. The planting of Bt cotton and disease-resistant papaya have provided efficient protection against cotton bollworms and Papaya ringspot virus (PRSV), respectively. As a consequence, chemical application to these crops has been significantly reduced, enhancing farm income while reducing human and non-target organism exposure to toxic chemicals. This article provides useful information for the colleagues, in particular for them whose mother tongue is not Chinese, to clearly understand the biosafety regulation and commercial use of genetically modified crops in China.

Does Expression of Bt Toxin Matter in Farmer's Pesticide Use?

Plant Biotechnology Journal. May, 2014  |  Pubmed ID: 24702829

Despite the widespread adoption of Bt cotton, farmers still spray excessive pesticides in their cotton fields. In contrast to scientists who always use high quality seeds in the laboratory and/or experimental fields, farmers may plant low quality seeds with a low expression of Bt toxin. How does the expression of Bt toxin influence farmers' pesticide use? On the basis of a plot-level survey and laboratory test data, this study shows that pesticide use on one cotton plot is influenced not only by the expression of Bt crops in this plot, but also by the average expression in the village in the early stage of the cotton growing season. In other words, high expression of Bt toxin benefits not only the farmers who plant the varieties but also all the other villagers.

Silencing DACH1 Promotes Esophageal Cancer Growth by Inhibiting TGF-β Signaling

PloS One. 2014  |  Pubmed ID: 24743895

Human Dachshund homologue 1 (DACH1) is a major component of the Retinal Determination Gene Network. Loss of DACH1 expression was found in breast, prostate, lung, endometrial, colorectal and hepatocellular carcinoma. To explore the expression, regulation and function of DACH1 in human esophageal cancer, 11 esophageal cancer cell lines, 10 cases of normal esophageal mucosa, 51 cases of different grades of dysplasia and 104 cases of primary esophageal squamous cancer were employed. Methylation specific PCR, immunohistochemistry, western blot, flow cytometry, small interfering RNAs, colony formation techniques and xenograft mice model were used. We found that DACH1 expression was regulated by promoter region hypermethylation in esophageal cancer cell lines. 18.8% (6 of 32) of grade 1, 42.1% (8 of 19) of grade 2 and grade 3 dysplasia (ED2,3), and 61.5% (64 of 104) of esophageal cancer were methylated, but no methylation was found in 10 cases of normal esophageal mucosa. The methylation was increased in progression tendency during esophageal carcinogenesis (P<0.01). DACH1 methylation was associated with poor differentiation (P<0.05) and late tumor stage (P<0.05). Restoration of DACH1 expression inhibited cell growth and activated TGF-β signaling in KYSE150 and KYSE510 cells. DACH1 suppressed human esophageal cancer cell tumor growth in xenograft mice. In conclusion, DACH1 is frequently methylated in human esophageal cancer and methylation of DACH1 is involved in the early stage of esophageal carcinogenesis. DACH1 expression is regulated by promoter region hypermethylation. DACH1 suppresses esophageal cancer growth by activating TGF-β signaling.

Plant Volatile Analogues Strengthen Attractiveness to Insect

PloS One. 2014  |  Pubmed ID: 24911460

Green leaf bug Apolygus lucorum (Meyer-Dür) is one of the major pests in agriculture. Management of A. lucorum was largely achieved by using pesticides. However, the increasing population of A. lucorum since growing Bt cotton widely and the increased awareness of ecoenvironment and agricultural product safety makes their population-control very challenging. Therefore this study was conducted to explore a novel ecological approach, synthetic plant volatile analogues, to manage the pest. Here, plant volatile analogues were first designed and synthesized by combining the bioactive components of β-ionone and benzaldehyde. The stabilities of β-ionone, benzaldehyde and analogue 3 g were tested. The electroantennogram (EAG) responses of A. lucorum adult antennae to the analogues were recorded. And the behavior assay and filed experiment were also conducted. In this study, thirteen analogues were acquired. The analogue 3 g was demonstrated to be more stable than β-ionone and benzaldehyde in the environment. Many of the analogues elicited EAG responses, and the EAG response values to 3 g remained unchanged during seven-day period. 3 g was also demonstrated to be attractive to A. lucorum adults in the laboratory behavior experiment and in the field. Its attractiveness persisted longer than β-ionone and benzaldehyde. This indicated that 3 g can strengthen attractiveness to insect and has potential as an attractant. Our results suggest that synthetic plant volatile analogues can strengthen attractiveness to insect. This is the first published study about synthetic plant volatile analogues that have the potential to be used in pest control. Our results will support a new ecological approach to pest control and it will be helpful to ecoenvironment and agricultural product safety.

Epigenetic Silencing of DACH1 Induces the Invasion and Metastasis of Gastric Cancer by Activating TGF-β Signalling

Journal of Cellular and Molecular Medicine. Dec, 2014  |  Pubmed ID: 24912879

Gastric cancer (GC) is the fourth most common malignancy in males and the fifth most common malignancy in females worldwide. DACH1 is frequently methylated in hepatic and colorectal cancer. To further understand the regulation and mechanism of DACH1 in GC, eight GC cell lines, eight cases of normal gastric mucosa, 98 cases of primary GC and 50 cases of adjacent non-tumour tissues were examined. Methylation-specific PCR, western blot, transwell assay and xenograft mice were used in this study. Loss of DACH1 expression correlated with promoter region methylation in GC cells, and re-expression was induced by 5-Aza-2'-deoxyazacytidine. DACH1 is methylated in 63.3% (62/98) of primary GC and 38% (19/50) of adjacent non-tumour tissues, while no methylation was found in normal gastric mucosa. Methylation of DACH1 correlated with reduced expression of DACH1 (P < 0.01), late tumour stage (stage III/IV) (P < 0.01) and lymph node metastasis (P < 0.05). DACH1 expression inhibited epithelial-mesenchymal transition and metastasis by inhibiting transforming growth factor (TGF)-β signalling and suppressed GC cell proliferation through inducing G2/M phase arrest. The tumour size is smaller in DACH1-expressed BGC823 cell xenograft mice than in unexpressed group (P < 0.01). Restoration of DACH1 expression also sensitized GC cells to docetaxel. These studies suggest that DACH1 is frequently methylated in human GC and expression of DACH1 was controlled by promoter region methylation. DACH1 suppresses GC proliferation, invasion and metastasis by inhibiting TGF-β signalling pathways both in vitro and in vivo. Epigenetic silencing DACH1 may induce GC cells' resistance to docetaxel.

Arthropod Abundance and Diversity in Transgenic Bt Soybean

Environmental Entomology. Aug, 2014  |  Pubmed ID: 24915416

Before the commercialization of any insect-resistant genetically modified crop, it must be subjected to a rigorous premarket risk assessment. Here, possible effects of growing of transgenic Cry1Ac soybean on arthropod communities under field conditions were assessed for 2 yr and quantified in terms of arthropod community indices including the Shannon-Weaver diversity index, richness index, and dominance index. Our results showed no significant differences of diversity, richness, or dominant indices for Bt soybean compared with the recipient cultivar, conventional soybean, or sprayed conventional soybean. Conventional soybean treatment with insecticide had an adverse effect on the arthropod community after spraying, but arthropod community diversity recovered quickly. Bt soybean had no negative effect on the dominant distribution of subcommunities, including sucking pests, other pests, predators, parasitoids, and others except for lepidopteran pests. The dominance distribution of lepidopteran pests decreased significantly in Bt soybean because of the significant decrease in the numbers of Spodoptera litura (F.) and Ascotis selenaria Schiffermüller et Denis compared with the recipient cultivar. Our results showed that there were no negative effects of Cry1Ac soybean on the arthropod community in soybean field plots in the short term.

Quantitative Analysis of Fitness Costs Associated with the Development of Resistance to the Bt Toxin Cry1Ac in Helicoverpa Armigera

Scientific Reports. Jul, 2014  |  Pubmed ID: 25005122

Transgenic Bacillus thuringiensis (Bt) crops play an increasing role in pest control, and resistance management is a major issue in large-scale cultivation of Bt crops. The fitness cost of resistance in targeted pests is considered to be one of the main factors delaying resistance when using the refuge strategy. By comparing 10 resistant Helicoverpa armigera (Hubner) strains, showing various resistance levels to Bt toxin (Cry1Ac), to a susceptible strain, we showed an increasing fitness cost corresponding with increasing levels of resistance. The relationship between overall fitness cost C and the resistance ratio Rr could be described by C = 24.47/(1 + exp([1.57 - Log10Rr]/0.2)). This model predicted that the maximum overall fitness cost would be ~24% (± 5.22) in the strains with the highest resistance level. The overall fitness cost was closely linked to egg hatching rate, fecundity, emergence rate, larval survival rate, and developmental duration of adults. Among fitness components measured, fecundity was the most sensitive trait linked to the resistance selection. To integrate the results into simulation models would be valuable in evaluating how variation in fitness cost may influence the development of resistance in pest populations, thus helping to develop enhanced refuge strategies.

Does Athetis Lepigone Moth (Lepidoptera: Noctuidae) Take a Long-distance Migration?

Journal of Economic Entomology. Jun, 2014  |  Pubmed ID: 25026658

Athetis lepigone (Möschler), a new lepidopteran pest in China, has spread quickly to seven provinces since it was first reported causing damage on summer maize in Hebei province in 2005, Whether this species is a migrant or not remains unknown. The past 3 yr searchlight trapping on an island in the center of Bohai Gulf provided direct evidence that both male and female A. lepigone moths migrate across the Bohai Gulf waters in northern China because no host crops or A. lepigone larvae were found on this island. The four migration waves observed in this study represent high-altitude movements of the overwintering, first, second, and third generations of A. lepigone moths, respectively. Carbon isotope analysis showed that 1.76-5.44% of the tested A. lepigone moths originated from C4 plants, which provides additional evidence that this species is a migrant because there are no C4 plants on this small island. The 89.24-96.89% of tested A. lepigone moths originated from C3 plants were significantly higher than that from C4 plants in all generations, suggesting that maize fields are not the main host sites for A. lepigone. Few females were trapped in spring and early summer with relatively high mating frequency and more advanced ovarian development, suggesting that the migration of this species is not completely bound by the "oogenesis-flight syndrome." These findings reveal a new route for A. lepigone migrating to and from the northeastern agricultural region of China, and improve our knowledge of the migration ecology of A. lepigone. Further studies are needed to clarify the migration trajectories that will help in developing sound forecasting systems for this pest species.

Seasonal Migration of Ctenoplusia Agnata (Lepidoptera: Noctuidae) over the Bohai Sea in Northern China

Journal of Economic Entomology. Jun, 2014  |  Pubmed ID: 25026659

Ctenoplusia agnata (Staudinger) (Lepidoptera: Noctuidae) is an important polyphagous pest in East Asia. Previous studies showed that C. agnata moths possesses the potential to undertake long-distance migration; however, knowledge of whether or not the migration of C. agnata moths is a regular ecological behavior and what the pattern of seasonal migrations is in case of regular migration is currently lacking. In the current study, systemic monitoring of population dynamics of C. agnata was conducted by a searchlight trap on an island in the center of Bohai gulf in northern China, during 2003-2013. Our results provided strong evidence for the hypothesis that C. agnata is one of the pest species undertaking regular high altitude long-distance migration and we have depicted the seasonal migration pattern over the Bohai Sea. The first capture of C. agnata generally appeared in late April and early May, then the daily number of catches increased to high levels in late July and formed two waves of migration through August and early September, and finally, the moths disappeared in late October. The mean time from the earliest trapping to the latest trapping within a year was 141.0 +/- 3.0 days. The index of ovarian development of female C. agnata showed seasonal variability and suggested that its migratory flight may be independent of the degree of ovarian development and mating status. In addition, strong migration events took place in 2003, 2004, 2008, and 2010 (annual sum of catches > 10,000). The research result from this work is helpful for understanding the occurrence regularity of C. agnata and developing an integrated pest management strategy.

Acquisition of Cry1Ac Protein by Non-target Arthropods in Bt Soybean Fields

PloS One. 2014  |  Pubmed ID: 25110881

Soybean tissue and arthropods were collected in Bt soybean fields in China at different times during the growing season to investigate the exposure of arthropods to the plant-produced Cry1Ac toxin and the transmission of the toxin within the food web. Samples from 52 arthropod species/taxa belonging to 42 families in 10 orders were analysed for their Cry1Ac content using enzyme-linked immunosorbent assay (ELISA). Among the 22 species/taxa for which three samples were analysed, toxin concentration was highest in the grasshopper Atractomorpha sinensis and represented about 50% of the concentration in soybean leaves. Other species/taxa did not contain detectable toxin or contained a concentration that was between 1 and 10% of that detected in leaves. These Cry1Ac-positive arthropods included a number of mesophyll-feeding Hemiptera, a cicadellid, a curculionid beetle and, among the predators, a thomisid spider and an unidentified predatory bug belonging to the Anthocoridae. Within an arthropod species/taxon, the Cry1Ac content sometimes varied between life stages (nymphs/larvae vs. adults) and sampling dates (before, during, and after flowering). Our study is the first to provide information on Cry1Ac-expression levels in soybean plants and Cry1Ac concentrations in non-target arthropods in Chinese soybean fields. The data will be useful for assessing the risk of non-target arthropod exposure to Cry1Ac in soybean.

Mis-splicing of the ABCC2 Gene Linked with Bt Toxin Resistance in Helicoverpa Armigera

Scientific Reports. Aug, 2014  |  Pubmed ID: 25154974

Toxins from the bacterium Bacillus thuringiensis (Bt) are used widely for insect control in sprays and transgenic plants, but their efficacy is reduced when pests evolve resistance. Previous work showed that mutations in a gene encoding the transporter protein ABCC2 are linked with resistance to Bt toxins Cry1Ab, Cry1Ac or both in four species of Lepidoptera. Here we compared the ABCC2 gene of Helicoverpa armigera (HaABCC2) between susceptible strains and a laboratory-selected strain with >1,000-fold resistance to Cry1Ac relative its susceptible parent strain. We discovered a 73-base pair (bp) insertion in the cDNA of the resistant strain that generates a premature stop codon expected to yield a truncated ABCC2 protein. Sequencing of genomic DNA revealed that this insertion is an intron that is not spliced out because of a 6-bp deletion at its splicing site. Analysis of progeny from crosses revealed tight genetic linkage between HaABCC2 and resistance to Cry1Ac. These results provide the first evidence that mis-splicing of a gene encoding an ABCC2 protein confers resistance to a Bt toxin.

Seasonal Migration of Apolygus Lucorum (Hemiptera: Miridae) over the Bohai Sea in Northern China

Journal of Economic Entomology. Aug, 2014  |  Pubmed ID: 25195428

During the past decade, Apolygus lucorum (Meyer-Dür) has become a key pest on cotton in northern China. Whether or not this species is a migrant, and if so, what pattern of seasonal migration this species exhibits remains unknown. The combination of searchlight trapping and radar observation on an isolated island in the center of Bohai Gulf during the past 11 yr provided direct evidence that both male and female A. lucorum adults migrate at least 40-60 km (and probably much greater distances) across the Bohai Gulf waters. There were considerable yearly and monthly variation in the number of A. lucorum trapped on BH, and the migration period during 2009-2013 ranged from 102 to 154 d. A. lucorum adults had downwind displacement rather than randomly by heading toward their seasonally favorable directions, which toward the east-northeast in summer, but south-southwest (SSW) in early autumn. The vast majority of adults flying at airspeeds 0.5-2.5 m/s and at altitudes < 150 m above ground level. Most of trapped females were virgins with little or no ovarian development, as suggests that the onset of migration is initiated mainly by sexually immature individuals, which is termed the "oogenesis-flight syndrome." Such findings reveal a new route for A. lucorum movements northward to and southward from the northeastern agricultural region of China, which will help us develop more effective management strategies against this pest species.

CAMK2N1 Inhibits Prostate Cancer Progression Through Androgen Receptor-dependent Signaling

Oncotarget. Nov, 2014  |  Pubmed ID: 25296973

Castration resistance is a major obstacle to hormonal therapy for prostate cancer patients. Although androgen independence of prostate cancer growth is a known contributing factor to endocrine resistance, the mechanism of androgen receptor deregulation in endocrine resistance is still poorly understood. Herein, the CAMK2N1 was shown to contribute to the human prostate cancer cell growth and survival through AR-dependent signaling. Reduced expression of CAMK2N1 was correlated to recurrence-free survival of prostate cancer patients with high levels of AR expression in their tumor. CAMK2N1 and AR signaling form an auto-regulatory negative feedback loop: CAMK2N1 expression was down-regulated by AR activation; while CAMK2N1 inhibited AR expression and transactivation through CAMKII and AKT pathways. Knockdown of CAMK2N1 in prostate cancer cells alleviated Casodex inhibition of cell growth, while re-expression of CAMK2N1 in castration-resistant cells sensitized the cells to Casodex treatment. Taken together, our findings suggest that CAMK2N1 plays a tumor suppressive role and serves as a crucial determinant of the resistance of prostate cancer to endocrine therapies.

DACH1 Inhibits Cyclin D1 Expression, Cellular Proliferation and Tumor Growth of Renal Cancer Cells

Journal of Hematology & Oncology. Oct, 2014  |  Pubmed ID: 25322986

Renal cell carcinoma (RCC) is a complex with diverse biological characteristics and distinct molecular signature. New target therapies to molecules that drive RCC initiation and progression have achieved promising responses in some patients, but the total effective rate is still far from satisfaction. Dachshund (DACH1) network is a key signaling pathway for kidney development and has recently been identified as a tumor suppressor in several cancer types. However, its role in renal cell carcinoma has not been fully investigated.

Densovirus is a Mutualistic Symbiont of a Global Crop Pest (Helicoverpa Armigera) and Protects Against a Baculovirus and Bt Biopesticide

PLoS Pathogens. Oct, 2014  |  Pubmed ID: 25357125

Mutualistic associations between symbiotic bacteria and their hosts are common within insect systems. However, viruses are often considered as pathogens even though some have been reported to be beneficial to their hosts. Herein, we report a novel densovirus, Helicoverpa armigera densovirus-1 (HaDNV-1) that appears to be beneficial to its host. HaDNV-1 was found to be widespread in wild populations of H. armigera adults (>67% prevalence between 2008 and 2012). In wild larval populations, there was a clear negative interaction between HaDNV-1 and H. armigera nucleopolyhedrovirus (HaNPV), a baculovirus that is widely used as a biopesticide. Laboratory bioassays revealed that larvae hosting HaDNV-1 had significantly enhanced resistance to HaNPV (and lower viral loads), and that resistance to Bacillus thuringiensis (Bt) toxin was also higher at low doses. Laboratory assays indicated that the virus was mainly distributed in the fat body, and could be both horizontally- and vertically-transmitted, though the former occurred only at large challenge doses. Densovirus-positive individuals developed more quickly and had higher fecundity than uninfected insects. We found no evidence for a negative effect of HaDNV-1 infection on H. armigera fitness-related traits, strongly suggesting a mutualistic interaction between the cotton bollworm and its densovirus.

Cis-mediated Down-regulation of a Trypsin Gene Associated with Bt Resistance in Cotton Bollworm

Scientific Reports. Nov, 2014  |  Pubmed ID: 25427690

Transgenic plants producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are useful for pest control, but their efficacy is reduced when pests evolve resistance. Here we examined the mechanism of resistance to Bt toxin Cry1Ac in the laboratory-selected LF5 strain of the cotton bollworm, Helicoverpa armigera. This strain had 110-fold resistance to Cry1Ac protoxin and 39-fold resistance to Cry1Ac activated toxin. Evaluation of five trypsin genes revealed 99% reduced transcription of one trypsin gene (HaTryR) was associated with resistance. Silencing of this gene with RNA interference in susceptible larvae increased their survival on diets containing Cry1Ac. Bioassays of progeny from crosses revealed that resistance to Cry1Ac was genetically linked with HaTryR. We identified mutations in the promoter region of HaTryR in the resistant strain. In transfected insect cell lines, transcription was lower when driven by the resistant promoter compared with the susceptible promoter, implicating cis-mediated down-regulation of HaTryR transcription as a mechanism of resistance. The results suggest that H. armigera can adapt to Bt toxin Cry1Ac by decreased expression of trypsin. Because trypsin activation of protoxin is a critical step in toxicity, transgenic plants with activated toxins rather than protoxins might increase the durability of Bt crops.

Notch Signaling and EMT in Non-small Cell Lung Cancer: Biological Significance and Therapeutic Application

Journal of Hematology & Oncology. Dec, 2014  |  Pubmed ID: 25477004

Through epithelial-mesenchymal transition (EMT), cancer cells acquire enhanced ability of migration and invasion, stem cell like characteristics and therapeutic resistance. Notch signaling regulates cell-cell connection, cell polarity and motility during organ development. Recent studies demonstrate that Notch signaling plays an important role in lung cancer initiation and cross-talks with several transcriptional factors to enhance EMT, contributing to the progression of non-small cell lung cancer (NSCLC). Correspondingly, blocking of Notch signaling inhibits NSCLC migration and tumor growth by reversing EMT. Clinical trials have showed promising effect in some cancer patients received treatment with Notch1 inhibitor. This review attempts to provide an overview of the Notch signal in NSCLC: its biological significance and therapeutic application.

Monitoring Cotton Bollworm Resistance to Cry1Ac in Two Counties of Northern China During 2009-2013

Pest Management Science. Mar, 2015  |  Pubmed ID: 24753356

Transgenic cotton that expresses a gene derived from the bacterium Bacillus thuringiensis (Bt) has been deployed for combating cotton bollworm in China since 1997. As a follow-up on research started in 2002, the quantitative shifts in larval Cry1Ac resistance of field Helicoverpa armigera populations were monitored from 2009-2013 using bioassays of isofemale lines.

Large-scale Test of the Natural Refuge Strategy for Delaying Insect Resistance to Transgenic Bt Crops

Nature Biotechnology. Feb, 2015  |  Pubmed ID: 25503384

The 'natural refuge strategy" for delaying insect resistance to transgenic cotton that produces insecticidal proteins from Bacillus thuringiensis (Bt) relies on refuges of host plants other than cotton that do not make Bt toxins. We tested this widely adopted strategy by comparing predictions from modeling with data from a four-year field study of cotton bollworm (Helicoverpa armigera) resistance to transgenic cotton producing Bt toxin Cry1Ac in six provinces of northern China. Bioassay data revealed that the percentage of resistant insects increased from 0.93% in 2010 to 5.5% in 2013. Modeling predicted that the percentage of resistant insects would exceed 98% in 2013 without natural refuges, but would increase to only 1.1% if natural refuges were as effective as non-Bt cotton refuges. Therefore, the results imply that natural refuges delayed resistance, but were not as effective as an equivalent area of non-Bt cotton refuges. The percentage of resistant insects with nonrecessive inheritance of resistance increased from 37% in 2010 to 84% in 2013. Switching to Bt cotton producing two or more toxins and integrating other control tactics could slow further increases in resistance.

Cross-resistance and Interactions Between Bt Toxins Cry1Ac and Cry2Ab Against the Cotton Bollworm

Scientific Reports. Jan, 2015  |  Pubmed ID: 25586723

To delay evolution of pest resistance to transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt), the "pyramid" strategy uses plants that produce two or more toxins that kill the same pest. We conducted laboratory diet experiments with the cotton bollworm, Helicoverpa armigera, to evaluate cross-resistance and interactions between two toxins in pyramided Bt cotton (Cry1Ac and Cry2Ab). Selection with Cry1Ac for 125 generations produced 1000-fold resistance to Cry1Ac and 6.8-fold cross-resistance to Cry2Ab. Selection with Cry2Ab for 29 generations caused 5.6-fold resistance to Cry2Ab and 61-fold cross-resistance to Cry1Ac. Without exposure to Bt toxins, resistance to both toxins decreased. For each of the four resistant strains examined, 67 to 100% of the combinations of Cry1Ac and Cry2Ab tested yielded higher than expected mortality, reflecting synergism between these two toxins. Results showing minor cross-resistance to Cry2Ab caused by selection with Cry1Ac and synergism between these two toxins against resistant insects suggest that plants producing both toxins could prolong the efficacy of Bt cotton against this pest in China. Including toxins against which no cross-resistance occurs and integrating Bt cotton with other control tactics could also increase the sustainability of management strategies.

The Endogenous Cell-fate Factor Dachshund Restrains Prostate Epithelial Cell Migration Via Repression of Cytokine Secretion Via a Cxcl Signaling Module

Cancer Research. May, 2015  |  Pubmed ID: 25769723

Prostate cancer is the second leading form of cancer-related death in men. In a subset of prostate cancer patients, increased chemokine signaling IL8 and IL6 correlates with castrate-resistant prostate cancer (CRPC). IL8 and IL6 are produced by prostate epithelial cells and promote prostate cancer cell invasion; however, the mechanisms restraining prostate epithelial cell cytokine secretion are poorly understood. Herein, the cell-fate determinant factor DACH1 inhibited CRPC tumor growth in mice. Using Dach1(fl/fl)/Probasin-Cre bitransgenic mice, we show IL8 and IL6 secretion was altered by approximately 1,000-fold by endogenous Dach1. Endogenous Dach1 is shown to serve as a key endogenous restraint to prostate epithelial cell growth and restrains migration via CXCL signaling. DACH1 inhibited expression, transcription, and secretion of the CXCL genes (IL8 and IL6) by binding to their promoter regulatory regions in chromatin. DACH1 is thus a newly defined determinant of benign and malignant prostate epithelium cellular growth, migration, and cytokine abundance in vivo.

Development of 10 Microsatellite Markers from Pantala Flavescens and Their Applicability in Studying Genetics Diversity

Molecular Biology Reports. Aug, 2015  |  Pubmed ID: 25788247

Pantala flavescens (Fabricius 1798) is one of the most common species among migration dragonflies. It is often encountered in large swarms during migration or directed dispersal flights. For a better understanding of its gene flow, genetic structure and migration patterns throughout the world, 10 polymorphic microsatellite markers were isolated in this study. We respectively collected 32 P. flavescens from three places (Hunan, Liaoning and Heilongjiang) and 20 P. flavescens from Beijing. Partial genomic libraries containing microsatellite sequences were constructed with magnetic-bead enrichment method. By screening, sequence analysis, PCR amplification and so on, ten 10 polymorphic microsatellite markers were isolated. In order to assess their applicability, genetic diversity of these novel markers was tested in 96 individuals from three populations in China (Hunan, Liaoning and Heilongjiang). These markers were highly polymorphic, with 3-12 alleles per markers. The observed (Ho) and expected (He) heterozygosities ranged 0.321-0.667 and from 0.531 to 0.948 respectively. The genetic difference between Hunan and Liaoning is 0.429, while the genetic difference between Liaoning and Heilongjiang is 0.0508. These microsatellite markers for P. flavescens were developed for the first time, and will be a powerful tool for studying population genetic diversity and dispersal behavior of P. flavescens in China and worldwide.

DACH1 Inhibits Lung Adenocarcinoma Invasion and Tumor Growth by Repressing CXCL5 Signaling

Oncotarget. Mar, 2015  |  Pubmed ID: 25788272

Whole-genome and transcriptome sequencing of non-small cell lung cancer (NSCLC) identified that DACH1, is a human homolog of drosophila gene dac, is involved in NSCLC. Here we showed that expression of DACH1 was significantly decreased in human NSCLC tissues and DACH1 abundance was inversely correlated with tumor stages and grades. Restoration of DACH1 expression in NSCLC cells significantly reduced cellular proliferation, clone formation, migration and invasion in vitro, as well as tumor growth in vivo. Unbiased screen and functional study suggested that DACH1 mediated effects were dependent in part on suppression of CXCL5. There was an inverse correlation between DACH1 mRNA levels and CXCL5 in both lung cancer cell lines and human NSCLC tissues. Kaplan-Mier analysis of human NSCLC samples demonstrated that high DACH1 mRNA levels predicted favorable prognosis for relapse-free and overall survival. In agreement, high CXCL5 expression predicted a worse prognosis for survival.

A Toxin-binding Alkaline Phosphatase Fragment Synergizes Bt Toxin Cry1Ac Against Susceptible and Resistant Helicoverpa Armigera

PloS One. 2015  |  Pubmed ID: 25885820

Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry) proteins from the bacterium Bacillus thuringiensis (Bt) in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50) of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f) was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects.

DACH1 is a Novel Predictive and Prognostic Biomarker in Hepatocellular Carcinoma As a Negative Regulator of Wnt/β-catenin Signaling

Oncotarget. Apr, 2015  |  Pubmed ID: 25940701

The cell fate determination factor Dachshund (DACH1) functions as a novel suppressor in the progression of various neoplasms. Previous study has suggested that hypermethylation of promoter region was responsible for the reduction of DACH1 expression in hepatocellular carcinoma (HCC), and associated with the progression of HCC, but the clinical significance and the exact molecular mechanisms of DACH1 in the progression of HCC remain unclear. In this study, we employed public microarray data analysis and tissue microarrays (TMAs) technologies and showed that DACH1 expression was reduced in HCC even at early stage and associated with the tumor progression. Notably, Kaplan-Meier analysis further indicated DACH1 could be an independent prognostic factor for the overall survival of HCC. Further, mechanistic studies revealed that overexpression of DACH1 inhibited the growth and migration of HCC cell line, which were dependent in part on the inactivation of Wnt pathway via phosphorylation of GSK3β to suppress β-catenin. In agreement, the abundance of DACH1 was inversely correlated with several Wnt target genes. Collectively, our study indicated β-catenin is a novel target of DACH1 in HCC.

Molecular Characterization and Expression Profiles of Polygalacturonase Genes in Apolygus Lucorum (Hemiptera: Miridae)

PloS One. 2015  |  Pubmed ID: 25955307

Polygalacturonase (PG) is an enzyme in the salivary glands of piercing-sucking mirid bugs (Hemiptera: Miridae) that plays a key role in plant feeding and injury. By constructing a full-length cDNA library, we cloned and characterized 14 PG genes from the salivary glands of Apolygus lucorum, a pestiferous mirid bug in cotton, fruit trees and other crops in China. BLAST search analysis showed that the amino acid sequences deduced from transcripts of the PG genes were closely related to PGs from other mirid bugs. Phylogenetic analysis showed that the PGs of mirid bugs had six main branches, PG1-PG6 (Genbank accession numbers: KF881899~KF881912). We investigated the mRNA expression patterns of the A. lucorum PG genes using real-time PCR. All 14 PGs were expressed significantly higher in the salivary glands than in other tissues (head, thorax, abdomen, leg and wing). For eggs and nymphs, the expression levels of these PGs were much higher in the 5th instar stage than in the egg, and 1st and 3rd instar stages. The PG expression levels in 1-day-old adults were very low, and increased in 5, 20 and 30-day-old adults. Additionally, PG expression levels were generally similar between males and females. The possible physiological functions of PGs in A. lucorum were discussed.

Meta-analysis Reveals the Correlation of Notch Signaling with Non-small Cell Lung Cancer Progression and Prognosis

Scientific Reports. May, 2015  |  Pubmed ID: 25996086

Various studies have assessed the clinicopathological and prognostic value of Notch1 and Notch3 expression in Non-small cell lung cancer (NSCLC), but their results remain controversial. This meta-analysis was conducted to address the above issues by using a total of 19 studies involving 3663 patients. The correlations between Notch1 and Notch3 expression and clinicopathological features and NSCLC prognosis were analyzed. The meta-analysis indicated that higher expression of Notch1 was associated with greater possibility of lymph node metastasis and higher TNM stages. Moreover, patients with Notch1 overexpression and Notch3 overexpression showed significantly poor overall survival (Notch1: HR, 1.29; 95% CI, 1.06-1.57, p = 0.468 and I(2) = 0.0%; Notch3: HR, 1.57; 95%CI, 1.04-2.36, p = 0.445 and I(2) = 0.0%). Furthermore, there are statistically significant association between overall survival of NSCLC patients and the expression of Notch signaling ligand DLL3 and target gene HES1. Our meta-analysis supports that Notch signaling is a valuable bio-marker to predict progression and targeting Notch signaling could benefit subpopulation of NSCLC patients.

Annual Migration of Agrotis Segetum (Lepidoptera: Noctuidae): Observed on a Small Isolated Island in Northern China

PloS One. 2015  |  Pubmed ID: 26114576

Migration behavior of the turnip moth, Agrotis segetum (Lepidoptera: Noctuidae), is not well known by far. Here, we present the data from an 11-year study on A. segetum by means of searchlight trapping and ovarian dissection on Beihuang (BH) Island, which located in the center of the Bohai Strait in northern China. The data showed a large number of A. segetum flight across the strait each year, which provides direct evidence that A. segetum is a long-distance migrant, migrating at least 40-60 km to reach the trapping site. The migration period during 2003-2013 ranged from 115 to 172 d. Among the catches, the proportion of females was significantly higher than that of males in each month from May to September. Ovarian dissection showed that the proportion of mated females and the proportion of sexually mature females was significantly higher than that of unmated females and sexually immature females in early summer, respectively, but conversely in autumn. The early summer populations migrate in a south-north direction, which might undertake a long-distance flight on several successive nights. The autumn populations migrate in a north-south direction, which might originate not far from the trapping site. Based on these findings, the migratory physiology of A. segetum was discussed.

Expression of Notch1 Correlates with Breast Cancer Progression and Prognosis

PloS One. 2015  |  Pubmed ID: 26121683

Various studies have evaluated the significance of Notch1 expression in breast cancer, but the results have ever been disputed. By using 21 studies involving 3867 patients, this meta-analysis revealed that the expression of Notch1 was significantly higher in breast cancer than in normal tissues (OR=7.21; 95%CI, 4.7-11.07) and that higher Notch1 expression was associated with transition from ductal carcinoma in situ (DCIS) to invasive cancer (OR=3.75; 95% CI, 1.8-7.78). Higher Notch1 activity was observed in the basal subtype of breast cancer (OR=2.53; 95% CI, 1.18-5.43). Moreover, patients with Notch1 overexpression exhibited significantly worse overall and recurrence-free survival. Our meta-analysis suggests that Notch inhibitors may be useful in blocking the early progression of DCIS and that the outcomes of clinical trials for Notch1-targeting therapeutics could be improved by the molecular stratification of breast cancer patients.

Annual Migration of Cabbage Moth, Mamestra Brassicae L. (Lepidoptera: Noctuidae), over the Sea in Northern China

PloS One. 2015  |  Pubmed ID: 26176951

The cabbage moth, Mamestra brassicae L. (Lepidoptera: Noctuidae), is a serious pest of vegetable crops throughout the world. In order to determine whether or not M. brassicae is a migrant, and if yes, what is the pattern of M. brassicae seasonal migration, a long-term study on M. brassicae from April to October in 2003-2014 was carried out by means of a searchlight trap on a small island located in the center of the Bohai Strait. The results show that a large number of M. brassicae were trapped every year on the island, which indicates that M. brassicae is a migrant and migrated at least 40-60 km across the Bohai Strait. The mean migration period of M. brassicae over the sea within one year is 151 ± 8 d in 2003-2014, with the shortest time span 78 d in 2003 and the longest 189 d in 2014, respectively. The number of M. brassicae captured, however, varies considerably between months or years. The majority of captures were female, with different levels of ovarian development and mating status. Most of the females trapped in May-July during 2010-2014 had a high mating rate and advanced level of ovarian development, suggesting that the migration of this species does not conform to the hypothesis of 'oogenesis-flight syndrome'. The findings of the present study are beneficial to the development of forecasting systems and management strategies of M. brassicae.

Non-invasive Approaches to Monitor EGFR-TKI Treatment in Non-small-cell Lung Cancer

Journal of Hematology & Oncology. Jul, 2015  |  Pubmed ID: 26227959

Tyrosine kinase inhibitors of epidermal growth factor receptor (EGFR-TKIs) are standard treatments for advanced non-small-cell lung cancer (NSCLC) patients harboring activating epidermal growth factor receptor (EGFR) mutations. Nowadays, tumor tissues acquired by surgery or biopsy are the routine materials for EGFR mutation analysis. However, the accessibility of tumor tissues is not always satisfactory in advanced NSCLC. Moreover, a high proportion of NSCLC patients will eventually develop resistance to EGFR-TKIs. Invasive procedures, such as surgery or biopsy, are impractical to be performed repeatedly to assess the evolution of EGFR-TKI resistance. Thus, exploring some convenient and less invasive techniques to monitor EGFR-TKI treatment is urgently needed. Circulating cell-free tumor DNA (ctDNA) has a high degree of specificity to detect EGFR mutations in NSCLC. Besides, ctDNA is capable of monitoring the disease progression during EGFR-TKI treatment. Certain serum microRNAs that correlate with EGFR signaling pathway, such as miR-21 and miR-10b, have been demonstrated to be helpful in evaluating the efficiency of EGFR-TKI therapeutics. A commercialized serum-based proteomic test, named VeriStrat test, has shown an outstanding ability to predict the clinical outcome of NSCLC patients receiving EGFR-TKIs. Analysis of EGFR mutations in circulating tumor cells (CTCs) is feasible, and CTCs represent a promising material to predict EGFR-TKI-treatment efficacy and resistance. These evidences suggested that non-invasive techniques based on serum or plasma samples had a great potential for monitoring EGFR-TKI treatment in NSCLC. In this review, we summarized these non-invasive approaches and considered their possible applications in EGFR-TKI-treatment monitoring.

The Inhibitory Effects of AR/miR-190a/YB-1 Negative Feedback Loop on Prostate Cancer and Underlying Mechanism

Scientific Reports. Aug, 2015  |  Pubmed ID: 26314494

Prostate cancer at advanced stages including metastatic and castration-resistant cancer remains incurable due to the lack of effective therapies. MiR-190a belongs to the small noncoding RNA family and has an important role in breast cancer metastasis. However, it is still unknown whether miR-190a plays a role in prostate cancer development. Herein, we first observed AR/miR-190a/YB-1 forms an auto-regulatory negative feedback loop in prostate cancer: miR-190a expression was down-regulated by AR activation; YB-1 functions are as an AR activator; miR-190a inhibited AR expression and transactivation through direct binding to 3'UTR of YB-1 gene. MiR-190a contributes the human prostate cancer cell growth through AR-dependent signaling. Moreover, we examined the expression of miR-190a and observed a significant decrease in human prostate cancers. Reduced expression of miR-190a was inversely correlated to AR levels of prostate cancer patients, and patients with higher miR-190a expression in their tumor have improved tumor-free survival. Taken together, our findings identified a biochemical and functional link between miR-190a with reduced expression in advanced prostate cancer, YB-1 and AR signaling in prostate cancer.

Notch Signaling: an Emerging Therapeutic Target for Cancer Treatment

Cancer Letters. Dec, 2015  |  Pubmed ID: 26341688

The Notch pathway is involved in cell proliferation, differentiation and survival. The Notch signaling pathway is one of the most commonly activated signaling pathways in cancer. Alterations include activating mutations and amplification of the Notch pathway, which play key roles in the progression of cancer. Accumulating evidence suggests that the pharmacological inhibition of this pathway can overcome chemoresistance. Efforts have been taken to develop Notch inhibitors as a single agent or in combination with clinically used chemotherapeutics to treat cancer. Some Notch inhibitors have been demonstrated to have therapeutic efficacy in preclinical studies. This review summarizes the recent studies and clinical evaluations of the Notch inhibitors in cancer.

Volatile Fragrances Associated with Flowers Mediate Host Plant Alternation of a Polyphagous Mirid Bug

Scientific Reports. 10, 2015  |  Pubmed ID: 26423224

Apolygus lucorum (Hemiptera: Miridae) is an important insect pest of cotton and fruit trees in China. The adults prefer host plants at the flowering stage, and their populations track flowering plants both spatially and temporally. In this study, we examine whether flower preference of its adults is mediated by plant volatiles, and which volatile compositions play an important role in attracting them. In olfactometer tests with 18 key host species, the adults preferred flowering plants over non-flowering plants of each species. Coupled gas chromatography-electroantennography revealed the presence of seven electrophysiologically active compounds from flowering plants. Although the adults responded to all seven synthetic plant volatiles in electroantennography tests, only four (m-xylene, butyl acrylate, butyl propionate and butyl butyrate) elicited positive behavioral responses in Y-tube olfactometer bioassays. The adults were strongly attracted to these four active volatiles in multi-year laboratory and field trials. Our results suggest that these four fragrant volatiles, which are emitted in greater amounts once plants begin to flower, mediate A. lucorum's preference to flowering host plants. We proved that the use of commonly occurring plant volatiles to recognize a large range of plant species can facilitate host selection and preference of polyphagous insect herbivore.

Seasonal Pattern of Spodoptera Litura (Lepidoptera: Noctuidae) Migration Across the Bohai Strait in Northern China

Journal of Economic Entomology. Apr, 2015  |  Pubmed ID: 26470163

The common cutworm, Spodoptera litura (F.), is a serious crop pest with a strong migratory ability. Previous studies on the migration of S. litura were mostly carried out in its "year-round breeding region" (YBR) or "overwintering region" (OR). However, the pattern of seasonal movements in its "summer breeding region" (SBR; i.e., northern China where they cannot overwinter) remains unknown. Here, we present data from an 11-yr study of this species made by searchlight trapping on Beihuang (BH) Island in the center of the Bohai Strait, which provides direct evidence that S. litura regularly migrates across this sea. There was considerable yearly and monthly variation in the number of S. litura trapped on BH, with the vast majority trapped in the autumn. The mean time from the earliest trapping to the latest trapping within a year was 110±12 d during 2003-2013, with the shortest time span of 40 d in 2003 and the longest of 166 d in 2012. S. litura moths had downwind displacement rather than randomly by heading toward their seasonally favorable direction (i.e. toward southwest in the four autumn migration events by prevailing northeasterly winds). Some females trapped in July showed a relatively higher proportion of having mated and a degree of ovarian development, suggesting that the migration of this species is not completely bound by the "oogenesis-flight syndrome." These findings provide a good starting point of research on S. litura migration between its OR (or YBR) and SBR, which will help us develop more effective regional management strategies against this pest.

Endogenous Dach1 in Cancer

Oncoscience. 2015  |  Pubmed ID: 26682253

The Role of CD44 in Epithelial-mesenchymal Transition and Cancer Development

OncoTargets and Therapy. 2015  |  Pubmed ID: 26719706

CD44, a multi-structural and multifunctional transmembrane glycoprotein, was initially identified as a receptor for hyaluronan that participates in both physiological and pathological processes. CD44 is found to be closely linked to the development of various solid tumors. Molecular studies have revealed that high CD44 expression was correlated with the phenotypes of cancer stem cells and epithelial-mesenchymal transition, thereby contributing to tumor invasion, metastasis, recurrence, and chemoresistance. Correspondingly, blockade of CD44 has been demonstrated to be capable of attenuating the malignant phenotype, slowing cancer progression, and reversing therapy resistance. Clinical analyses showed that high CD44 expression is associated with poor survival of various cancer patients, indicating that CD44 can be a potential prognostic marker. In this review, we summarize recent research progress of CD44 on tumor biology and the clinical significance of CD44.

Interplay of Retinal Determination Gene Network with TGF-β Signaling Pathway in Epithelial-mesenchymal Transition

Stem Cell Investigation. 2015  |  Pubmed ID: 27358880

As a fundamental event in the generation of tissues and organs during embryogenesis, the epithelial-mesenchymal transition (EMT) has also been implicated in cancer progression by its ability to alter the plasticity of epithelial cells to acquire invasive properties. Evidence is mounting that ectopic activation of transforming growth factors β (TGF-β)/bone morphogenetic protein (BMP) superfamily members to enhance tumorigenesis and metastasis. In this respect, the Retinal Determination Gene Network (RDGN), which was identified to govern the normal initiation of the morphogenetic furrow in Drosophila, has now been found to be de-regulated in various types of cancers, and the key members of this network, DACH, SIX, and EYA, have emerged as novel co-regulators of TGF- signaling during EMT. Understanding the molecular mechanism by which RDGN regulates TGF-β/BMP signaling to influence EMT may lead to novel strategies for targeted therapies.

The DACH/EYA/SIX Gene Network and Its Role in Tumor Initiation and Progression

International Journal of Cancer. Mar, 2016  |  Pubmed ID: 26096807

The functional abnormality of developmental genes is a common phenomenon in cancer initiation and progression. The retinal determination gene network (RDGN) is a key signal in Drosophila eye specification, and this conservative pathway is also required for the development of multiple organs in mammalian species. Recent studies demonstrated that aberrant expressions of RDGN components in vertebrates, mainly Dach, Six, and Eya, represent a novel tumor signal. RDGN regulates proliferation, apoptosis, tumor growth and metastasis through interactions with multiple signaling pathways in a co-ordinated fashion; Dach acts as a tumor suppressor, whereas Six and Eya function as oncogenes. Clinical analyses demonstrated that the expression levels of RDGN correlate with tumor stage, metastasis and survival, suggesting that combinational detection of this pathway might be used as a promising biomarker for the stratification of therapy and for the prediction of the prognosis of cancer patients.

The Development and Status of Bt Rice in China

Plant Biotechnology Journal. Mar, 2016  |  Pubmed ID: 26369652

Multiple lines of transgenic rice expressing insecticidal genes from the bacterium Bacillus thuringiensis (Bt) have been developed in China, posing the prospect of increases in production with decreased application of pesticides. We explore the issues facing adoption of Bt rice for commercial production in China. A body of safety assessment work on Bt rice has shown that Bt rice poses a negligible risk to the environment and that Bt rice products are as safe as non-Bt control rice products as food. China has a relatively well-developed regulatory system for risk assessment and management of genetically modified (GM) plants; however, decision-making regarding approval of commercial production has become politicized, and two Bt rice lines that otherwise were ready have not been allowed to enter the Chinese agricultural system. We predict that Chinese farmers would value the prospect of increased yield with decreased use of pesticide and would readily adopt production of Bt rice. That Bt rice lines may not be commercialized in the near future we attribute to social pressures, largely due to the low level of understanding and acceptance of GM crops by Chinese consumers. Hence, enhancing communication of GM crop science-related issues to the public is an important, unmet need. While the dynamics of each issue are particular to China, they typify those in many countries where adoption of GM crops has been not been rapid; hence, the assessment of these dynamics might inform resolution of these issues in other countries.

APN1 is a Functional Receptor of Cry1Ac but Not Cry2Ab in Helicoverpa Zea

Scientific Reports. Jan, 2016  |  Pubmed ID: 26755166

Lepidopteran midgut aminopeptidases N (APNs) are phylogenetically divided into eight clusters, designated as APN1-8. Although APN1 has been implicated as one of the receptors for Cry1Ac in several species, its potential role in the mode of action of Cry2Ab has not been functionally determined so far. To test whether APN1 also acts as one of the receptors for Cry1Ac in Helicoverpa zea and even for Cry2Ab in this species, we conducted a gain of function analysis by heterologously expressing H. zea APN1 (HzAPN1) in the midgut and fat body cell lines of H. zea and the ovarian cell line of Spodoptera frugiperda (Sf9) and a loss of function analysis by RNAi (RNA interference) silencing of the endogenous APN1 in the three cell lines using the HzAPN1 double strand RNA (dsRNA). Heterologous expression of HzAPN1 significantly increased the susceptibility of the three cell lines to Cry1Ac, but had no effects on their susceptibility to Cry2Ab. Knocking down of the endogenous APN1 made the three cell lines resistant to Cry1Ac, but didn't change cell lines susceptibility to Cry2Ab. The findings from this study demonstrate that HzAPN1 is a functional receptor of Cry1Ac, but not Cry2Ab.

GRIM-19 Inhibition Induced Autophagy Through Activation of ERK and HIF-1α Not STAT3 in Hela Cells

Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine. Jul, 2016  |  Pubmed ID: 26810068

Gene associated with retinoid-interferon-induced mortality (GRIM-19), an important subunit of mitochondrial complex I, has been identified as a tumor suppressor, and its reduced expression has been reported to be associated with tumorigenesis and metastasis. Autophagy has been proposed as a protective mechanism for cell survival under various stresses, including chemotherapy. However, it remains unknown whether GRIM-19 is linked to autophagy and chemotherapy resistance. Here, we showed that suppression of GRIM-19 by shRNA enhanced cell-type-dependent autophagy by activating extracellular regulated protein kinase (ERK) and hypoxia inducible factor-1a (HIF-1a) in a reactive oxygen species (ROS)-mediated manner, and thereby conferred resistance to paclitaxel. Besides, the antioxidant N-acetyl-L-cysteine (NAC) and autophagy inhibitor 3-MA could in part overcome this resistance. We also found that GRIM-19 expression was significantly correlated with clinical stage and grade in patients with cervical cancers. Taken together, our results indicated that GRIM-19 inhibition induced autophagy and chemotherapy resistance, which could affect prognosis of cervical cancers. Our study has identified new function of GRIM-19 and its underlying mechanism, and it will provide possible avenues for therapeutic targeting in cervical cancers.

Enrichment of CD44 in Basal-type Breast Cancer Correlates with EMT, Cancer Stem Cell Gene Profile, and Prognosis

OncoTargets and Therapy. 2016  |  Pubmed ID: 26855592

Cluster of differentiation 44 (CD44) is a transmembrane glycoprotein that serves as the receptor for the extracellular matrix component hyaluronic acid. CD44 has been reported to play key roles in cell proliferation, motility, and survival, but its role in breast cancer remains controversial. In this study, we conducted a meta-analysis. A total of 23 published Gene Expression Omnibus databases were included to evaluate the association between CD44 mRNA expression and clinicopathological characteristics or prognosis of the patients with breast cancer. Our analysis revealed that CD44 expression was associated with clinicopathological features, including the histological grade, estrogen receptor status, progesterone receptor status, and human epidermal growth factor receptor-2 status. Higher levels of CD44 expression were observed in the basal subtype of breast cancer both at the mRNA and protein levels (odds ratio [OR] =2.08, 95% confidence interval [CI]: 1.72-2.52; OR =2.11, 95% CI: 1.67-2.68). Patients with CD44 overexpression exhibited significantly worse overall survival (hazard ratio =1.27; 95% CI: 1.04-1.55). Whole gene profile analysis revealed that CD44 expression was enriched in basal-type breast cancer and correlated with epithelial-mesenchymal transition and cancer stem cell gene profiles. In summary, our analyses indicated that CD44 potentially might be a prognostic marker for breast cancer and thus can serve as a therapeutic target for basal-type breast cancer.

Resistance to Bacillus Thuringiensis Mediated by an ABC Transporter Mutation Increases Susceptibility to Toxins from Other Bacteria in an Invasive Insect

PLoS Pathogens. Feb, 2016  |  Pubmed ID: 26872031

Evolution of pest resistance reduces the efficacy of insecticidal proteins from the gram-positive bacterium Bacillus thuringiensis (Bt) used widely in sprays and transgenic crops. Recent efforts to delay pest adaptation to Bt crops focus primarily on combinations of two or more Bt toxins that kill the same pest, but this approach is often compromised because resistance to one Bt toxin causes cross-resistance to others. Thus, integration of Bt toxins with alternative controls that do not exhibit such cross-resistance is urgently needed. The ideal scenario of negative cross-resistance, where selection for resistance to a Bt toxin increases susceptibility to alternative controls, has been elusive. Here we discovered that selection of the global crop pest, Helicoverpa armigera, for >1000-fold resistance to Bt toxin Cry1Ac increased susceptibility to abamectin and spineotram, insecticides derived from the soil bacteria Streptomyces avermitilis and Saccharopolyspora spinosa, respectively. Resistance to Cry1Ac did not affect susceptibility to the cyclodiene, organophospate, or pyrethroid insecticides tested. Whereas previous work demonstrated that the resistance to Cry1Ac in the strain analyzed here is conferred by a mutation disrupting an ATP-binding cassette protein named ABCC2, the new results show that increased susceptibility to abamectin is genetically linked with the same mutation. Moreover, RNAi silencing of HaABCC2 not only decreased susceptibility to Cry1Ac, it also increased susceptibility to abamectin. The mutation disrupting ABCC2 reduced removal of abamectin in live larvae and in transfected Hi5 cells. The results imply that negative cross-resistance occurs because the wild type ABCC2 protein plays a key role in conferring susceptibility to Cry1Ac and in decreasing susceptibility to abamectin. The negative cross-resistance between a Bt toxin and other bacterial insecticides reported here may facilitate more sustainable pest control.

Emerging Roles of Nrf2 Signal in Non-small Cell Lung Cancer

Journal of Hematology & Oncology. Feb, 2016  |  Pubmed ID: 26922479

Non-small cell lung cancer (NSCLC) causes considerable mortality in the world. Owing to molecular biological progress, treatments in adenocarcinoma have evolved revolutionarily while those in squamous lung cancer remain unsatisfied. Recent studies revealed high-frequency alteration of Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-like factor 2 (Keap1/Nrf2) pathway within squamous lung cancer, attracting researchers to focus on this particular pathway. In NSCLC patients, deregulated Nrf2 signal is recognized as a common feature at both DNA and protein level. Emerging associations between Nrf2 and other pathways have been elucidated. MicroRNA was also implicated in the regulation of Nrf2. Agents activating or antagonizing Nrf2 showed an effect in preclinical researches, reflecting different effects of Nrf2 during tumor initiation and progression. Prognostic evaluation demonstrated a negative impact of Nrf2 signal on NSCLC patients' survival. Considering the importance of Nrf2 signal in NSCLC, further studies are required in the future.

Characterization of the Natural Enemy Community Attacking Cotton Aphid in the Bt Cotton Ecosystem in Northern China

Scientific Reports. Apr, 2016  |  Pubmed ID: 27075171

Planting Bt cotton in China since 1997 has led to important changes in the natural enemy communities occurring in cotton, however their specific effect on suppressing the cotton aphids (being notorious in conventional cotton ecosystem) has not been fully documented yet. We observed strong evidence for top-down control of the aphid population, e.g. the control efficiency of natural enemies on cotton aphid increased significantly in open field cages compared to exclusion cages, accounted for 60.2, 87.2 and 76.7% in 2011, 2012 and 2013 season, respectively. The cotton aphid populations peaked in early June to late July (early and middle growth stages) in open field cotton survey from 2011 to 2013. The population densities of cotton aphids and natural enemies were highest on middle growth stage while lowest densities were recorded on late stage for aphids and on early plant stage for natural enemies. Aphid parasitoids (Trioxys spp., Aphidius gifuensis), coccinellids and spiders were key natural enemies of cotton aphid. Briefly, natural enemies can suppress aphid population increase from early to middle plant growth stages by providing biocontrol services in Chinese Bt cotton.

Adult Exposure to Bt Toxin Cry1Ac Reduces Life Span and Reproduction of Resistant and Susceptible Pink Bollworm (Lepidoptera: Gelechiidae)

Journal of Economic Entomology. May, 2016  |  Pubmed ID: 27133579

Insecticidal proteins from Bacillus thuringiensis (Bt) are used widely in sprays and transgenic plants to control insect pests. Although much research has elucidated the effects of Bt toxins on larvae, relatively little is known about their effects on adults. Here, we evaluated the effects of exposing adults to Bt toxin Cry1Ac on the life span and reproduction of two strains of pink bollworm (Pectinophora gossypiella (Saunders)). In larval diet bioassays, the concentration of Cry1Ac killing 50% of larvae (LC50) was 640 times higher for the laboratory-selected resistant strain (AZP-R) than the susceptible strain (APHIS-S). In experiments with adults, the highest concentrations of Cry1Ac tested (160 and 640 µg Cry1Ac per ml of 5% honey water) reduced life span for both strains. Treatments with 10, 40, and 160 µg Cry1Ac per ml reduced the duration of the oviposition period as well as the number of eggs laid by both strains, but did not affect the percentage of pairs producing eggs, the duration of the preoviposition period, or the percentage of eggs hatching for either strain. Adult life span did not differ between strains at low to moderate concentrations of Cry1Ac, but it was significantly greater for the resistant strain than the susceptible strain at the two highest concentrations of Cry1Ac tested. The reduced susceptibility to high concentrations of Cry1Ac in adults of the AZP-R strain relative to the APHIS-S strain provides the first evidence of expression of resistance to a Bt toxin in adult Lepidoptera.

Molecular Cloning, Expression, and Identification of Bre Genes Involved in Glycosphingolipids Synthesis in Helicoverpa Armigera (Lepidoptera: Noctuidae)

Journal of Economic Entomology. May, 2016  |  Pubmed ID: 27190043

Glycosphingolipids (GSLs) play important roles in the cellular biology of vertebrate and invertebrate organisms, such as cell differentiation, tumor metastasis, and cell coordination. GSLs also serve as receptors for different bacterial toxins. For example, in the nematode Caenorhabditis elegans, GSLs function as receptors of the insecticidal Cry toxins produced by Bacillus thuringiensis (Bt), and mutations in bre genes involved in GSLs synthesis resulted in resistance to Cry5 toxin in this organism. However, the information of GSLs function in insects is still limited. In this study, three genes for glycosyltransferases, bre2, bre3, and bre4, from Helicoverpa armigera were identified and cloned. The previously reported bre5 gene from H. armigera was also analyzed. Protein sequence alignments revealed that proteins codified by H. armigera Bre shared high identity with homologous proteins from other organisms. Expression profile analysis revealed that the expressions of bre genes varied in the different tissues and also in the different developmental stages of H. armigera. Finally, the heterologous expression of bre genes in Trichoplusia ni Hi5 cell line showed that the corresponding translated proteins were localized in the cytoplasm of Hi5 cells. These results provide the bases for further functional studies of bre genes and analyzing potential roles of GSLs in mode of action of Cry1A toxin in H. armigera.

The Retinal Determination Gene Network: from Developmental Regulator to Cancer Therapeutic Target

Oncotarget. Aug, 2016  |  Pubmed ID: 27203207

Although originally identified for its function in Drosophila melanogaster eye specification, the Retinal Determination Gene Network (RDGN) is essential for the development of multiple organs in mammals. The RDGN regulates proliferation, differentiation and autocrine signaling, and interacts with other key signaling pathways. Aberrant expression of RDGN members such as DACH, EYA and SIX contributes to tumor initiation and progression; indeed, the levels of RDGN members are clinically prognostic factors in various cancer types. Stimulation or suppression of the activities of these crucial components can block cancer cell proliferation, prevent cancer stem cell expansion and even reverse the EMT process, thereby attenuating malignant phenotypes. Thus, cancer therapeutic interventions targeting RDGN members should be pursued in future studies.

Activation of Bt Protoxin Cry1Ac in Resistant and Susceptible Cotton Bollworm

PloS One. 2016  |  Pubmed ID: 27257885

Crystalline (Cry) proteins from Bacillus thuringiensis (Bt) are used extensively for insect control in sprays and transgenic plants, but their efficacy is reduced by evolution of resistance in pests. Here we evaluated reduced activation of Cry1Ac protoxin as a potential mechanism of resistance in the invasive pest Helicoverpa armigera. Based on the concentration killing 50% of larvae (LC50) for a laboratory-selected resistant strain (LF120) divided by the LC50 for its susceptible parent strain (LF), the resistance ratio was 1600 for Cry1Ac protoxin and 1200 for trypsin-activated Cry1Ac toxin. The high level of resistance to activated toxin as well as to protoxin indicates reduced activation of protoxin is not a major mechanism of resistance to Cry1Ac in LF120. For both insect strains, treatment with either the trypsin inhibitor N-a-tosyl-L-lysine chloromethyl ketone (TLCK) or the chymotrypsin inhibitor N-a-tosyl-L-phenylalanine chloromethyl ketone (TPCK) did not significantly affect the LC50 of Cry1Ac protoxin. Enzyme activity was higher for LF than LF120 for trypsin-like proteases, but did not differ between strains for chymotrypsin-like proteases. The results here are consistent with previous reports indicating that reduced activation of protoxin is generally not a major mechanism of resistance to Bt proteins.

Host Plants Identification for Adult Agrotis Ipsilon, a Long-Distance Migratory Insect

International Journal of Molecular Sciences. Jun, 2016  |  Pubmed ID: 27271592

In this study, we determined the host relationship of Agrotis ipsilon moths by identifying pollen species adhering them during their long-distance migration. Pollen carried by A. ipsilon moths was collected from 2012 to 2014 on a small island in the center of the Bohai Strait, which is a seasonal migration pathway of this pest species. Genomic DNA of single pollen grains was amplified by using whole genome amplification technology, and a portion of the chloroplast rbcL sequence was then amplified from this material. Pollen species were identified by a combination of DNA barcoding and pollen morphology. We found 28 species of pollen from 18 families on the tested moths, mainly from Angiosperm, Dicotyledoneae. From this, we were able to determine that these moths visit woody plants more than herbaceous plants that they carry more pollen in the early and late stages of the migration season, and that the amounts of pollen transportation were related to moth sex, moth body part, and plant species. In general, 31% of female and 26% of male moths were found to be carrying pollen. Amounts of pollen on the proboscis was higher for female than male moths, while the reverse was true for pollen loads on the antennae. This work provides a new approach to study the interactions between noctuid moth and their host plants. Identification of plant hosts for adult moths furthers understanding of the coevolution processes between moths and their host plants.

A Novel Paclitaxel-loaded Poly(d,l-lactide-co-glycolide)-Tween 80 Copolymer Nanoparticle Overcoming Multidrug Resistance for Lung Cancer Treatment

International Journal of Nanomedicine. 2016  |  Pubmed ID: 27307727

Drug resistance has become a main obstacle for the effective treatment of lung cancer. To address this problem, a novel biocompatible nanoscale package, poly(d,l-lactide-co-glycolide)-Tween 80, was designed and synthesized to overcome paclitaxel (PTX) resistance in a PTX-resistant human lung cancer cell line. The poly(d,l-lactide-co-glycolide) (PLGA)-Tween 80 nanoparticles (NPs) could efficiently load PTX and release the drug gradually. There was an increased level of uptake of PLGA-Tween 80 in PTX-resistant lung cancer cell line A549/T, which achieved a significantly higher level of cytotoxicity than both PLGA NP formulation and Taxol(®). The in vivo antitumor efficacy also showed that PLGA-Tween 80 NP was more effective than Taxol(®), indicating that PLGA-Tween 80 copolymer was a promising carrier for PTX in resistant lung cancer.

Left Lower Limb May Be a Forbidden Region for Indwelling Needle During Operation

Thrombosis Research. Aug, 2016  |  Pubmed ID: 27352239

Information about the incidence and risk factors of venous thromboembolism (VTE) after thyroid operation is limited. This study aimed to analyze the incidence and risk factors of postoperative VTE in patients who had undergone thyroid surgery.

Distribution and Metabolism of Bt-Cry1Ac Toxin in Tissues and Organs of the Cotton Bollworm, Helicoverpa Armigera

Toxins. Jul, 2016  |  Pubmed ID: 27399776

Crystal (Cry) proteins derived from Bacillus thuringiensis (Bt) have been widely used in transgenic crops due to their toxicity against insect pests. However, the distribution and metabolism of these toxins in insect tissues and organs have remained obscure because the target insects do not ingest much toxin. In this study, several Cry1Ac-resistant strains of Helicoverpa armigera, fed artificial diets containing high doses of Cry1Ac toxin, were used to investigate the distribution and metabolism of Cry1Ac in their bodies. Cry1Ac was only detected in larvae, not in pupae or adults. Also, Cry1Ac passed through the midgut into other tissues, such as the hemolymph and fat body, but did not reach the larval integument. Metabolic tests revealed that Cry1Ac degraded most rapidly in the fat body, followed by the hemolymph, peritrophic membrane and its contents. The toxin was metabolized slowly in the midgut, but was degraded in all locations within 48 h. These findings will improve understanding of the functional mechanism of Bt toxins in target insects and the biotransfer and the bioaccumulation of Bt toxins in arthropod food webs in the Bt crop ecosystem.

CD44 Correlates with Clinicopathological Characteristics and is Upregulated by EGFR in Breast Cancer

International Journal of Oncology. Oct, 2016  |  Pubmed ID: 27499099

Cluster of differentiation 44 (CD44), a well-known transmembrane glycoprotein, serves as a promoting factor in the carcinogenesis and progression of a variety of neoplasms. Previous studies have demonstrated that aberrant expression of CD44 was associated with the initiation, invasion, metastasis, and therapy-resistance of breast cancer, but whether there was any association between CD44 and pathological characteristics of breast cancer or epidermal growth factor receptor (EGFR) has not been clearly elucidated. In this study, we utilized public microarray data analysis and tissue microarray technologies to display that CD44 level was enhanced in breast cancer and was significantly correlated with histological grade and the status of estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2 (HER2) and EGFR. Furthermore, mRNA expression of CD44 in breast tumors was positively correlated with basal cytokeratin markers KRT5 and KRT17, but inversely associated with luminal marker FOXA1. Besides, Kaplan-Meier analysis showed that high CD44 mRNA level had adverse impact on the progression-free survival of patients with HER2-expressing or basal-like breast cancer. Functionally, inhibition of EGFR activity by erlotinib impaired the invasion and migration ability of breast cancer cell lines. Western blot assays demonstrated that erlotinib treatment decreased the expression of CD44, accompanied with the reduced protein levels of mesenchymal and cancer stem cell markers. Collectively, this study suggested that the expression of CD44 was upregulated by EGFR pathway and CD44 had a robust impact on the development of breast cancer.

The CXCL8-CXCR1/2 Pathways in Cancer

Cytokine & Growth Factor Reviews. Oct, 2016  |  Pubmed ID: 27578214

Persistent infection or chronic inflammation contributes significantly to tumourigenesis and tumour progression. C-X-C motif ligand 8 (CXCL8) is a chemokine that acts as an important multifunctional cytokine to modulate tumour proliferation, invasion and migration in an autocrine or paracrine manner. Studies have suggested that CXCL8 and its cognate receptors, C-X-C chemokine receptor 1 (CXCR1) and C-X-C chemokine receptor 2 (CXCR2), mediate the initiation and development of various cancers including breast cancer, prostate cancer, lung cancer, colorectal carcinoma and melanoma. CXCL8 also integrates with multiple intracellular signalling pathways to produce coordinated effects. Neovascularisation, which provides a basis for fostering tumour growth and metastasis, is now recognised as a critical function of CXCL8 in the tumour microenvironment. In this review, we summarize the biological functions and clinical significance of the CXCL8 signalling axis in cancer. We also propose that CXCL8 may be a potential therapeutic target for cancer treatment.

Modification of Platinum Sensitivity by KEAP1/NRF2 Signals in Non-small Cell Lung Cancer

Journal of Hematology & Oncology. Sep, 2016  |  Pubmed ID: 27601007

The objective of this study was to evaluate the effect of platinum-based drugs on nuclear-factor erythroid2 like 2 (NRF2) signaling in non-small cell lung cancer cell lines with or without Kelch-like ECH-associated protein 1 (KEAP1) mutations and to determine the role of NRF2 and KEAP1 on platinum-based drug treatment.

Functional Opsin Retrogene in Nocturnal Moth

Mobile DNA. 2016  |  Pubmed ID: 27777631

Retrotransposed genes are different to other types of genes as they originate from a processed mRNA and are then inserted back into the genome. For a long time, the contribution of this mechanism to the origin of new genes, and hence to the evolutionary process, has been questioned as retrogenes usually lose their regulatory sequences upon insertion and generally decay into pseudogenes. In recent years, there is growing evidence, notably in mammals, that retrotransposition is an important process driving the origin of new genes, but the evidence in insects remains largely restricted to a few model species.

The Expression Profile and Clinic Significance of the SIX Family in Non-small Cell Lung Cancer

Journal of Hematology & Oncology. Nov, 2016  |  Pubmed ID: 27821176

The SIX family homeobox genes have been demonstrated to be involved in the tumor initiation and progression, but their clinicopathological features and prognostic values in non-small cell lung cancer (NSCLC) have not been well defined. We analyzed relevant datasets and performed a systemic review and a meta-analysis to assess the profile of SIX family members in NSCLC and evaluate their importance as biomarkers for diagnosis and prediction of NSCLC.

OK-432 (Sapylin) Reduces Seroma Formation After Axillary Lymphadenectomy in Breast Cancer

Journal of Investigative Surgery : the Official Journal of the Academy of Surgical Research. Feb, 2017  |  Pubmed ID: 27431576

Purpose/aim: Modified radical mastectomy is the standard surgery for breast cancer in developing countries. However, seroma formation regarded as the most frequent postoperative complication limits the therapeutic benefit of mastectomy and axillary surgery. The purpose of this study was to evaluate the efficacy of OK-432 in reducing seroma formation after axillary dissection.

Activities of Digestive Enzymes in the Omnivorous Pest Apolygus Lucorum (Hemiptera: Miridae)

Journal of Economic Entomology. Feb, 2017  |  Pubmed ID: 28039425

The mirid bug Apolygus lucorum Meyer-Dür, 1843, an omnivorous species that feeds on plants and animals, has become a major pest in China as production of Bt-cotton has grown to such a large scale. Its omnivory is likely to be critical for its success, but the digestive mechanism(s) underlying processing and adsorption of such diverse foods is relatively unknown. Here, we examined the activities of digestive enzymes of A. lucorum in the salivary gland complex and midgut and the effect of sex, age, and food source on these activities. Amylase and protease were present in the salivary gland complex and the midgut, but were higher in the salivary gland complex. Trypsin-like enzyme was also present in both organs, but chymotrypsin-like enzyme was present only in the midgut. Sex, age, and food source affected the activities of these digestive enzymes. In general, the activities of these enzymes peaked at 10 d after emergence, and amylase and protease activities were higher in female adults than in males. Of the food sources tested, green bean pods (Gb) induced the highest amylase activity, whereas Helicoverpa armigera Hübner, 1809 eggs (He) and a mixture of Gb and He induced higher activities of the trypsin-like and chymotrypsin-like enzymes. The results from food switching experiments confirmed that amylase activity could be induced by plant sources, and animal sources induced protease activity. Thus, the types and activities of digestive enzymes in A. lucorum provide the physiological basis of the pest's omnivory.

Prostate-specific IL-6 Transgene Autonomously Induce Prostate Neoplasm Through Amplifying Inflammation in the Prostate and Peri-prostatic Adipose Tissue

Journal of Hematology & Oncology. Jan, 2017  |  Pubmed ID: 28077171

The causative role of the pro-inflammatory cytokine IL-6 in prostate cancer progression has been well established at molecular level. However, whether and how IL-6 may play a role in prostate cancer risk and development is not well defined. One limitation factor to acquiring this knowledge is the lack of appropriate animal models.

A Single Point Mutation Resulting in Cadherin Mislocalization Underpins Resistance Against Bacillus Thuringiensis Toxin in Cotton Bollworm

The Journal of Biological Chemistry. Feb, 2017  |  Pubmed ID: 28082675

Transgenic plants that produce Bacillus thuringiensis (Bt) crystalline (Cry) toxins are cultivated worldwide to control insect pests. Resistance to B. thuringiensis toxins threatens this technology, and although different resistance mechanisms have been identified, some have not been completely elucidated. To gain new insights into these mechanisms, we performed multiple back-crossing from a 3000-fold Cry1Ac-resistant BtR strain from cotton bollworm (Helicoverpa armigera), isolating a 516-fold Cry1Ac-resistant strain (96CAD). Cry1Ac resistance in 96CAD was tightly linked to a mutant cadherin allele (mHaCad) that contained 35 amino acid substitutions compared with HaCad from a susceptible strain (96S). We observed significantly reduced levels of the mHaCad protein on the surface of the midgut epithelium in 96CAD as compared with 96S. Expression of both cadherin alleles from 96CAD and 96S in insect cells and immunofluorescence localization in insect midgut tissue sections showed that the HaCAD protein from 96S localizes on the cell membrane, whereas the mutant 96CAD-mHaCad was retained in the endoplasmic reticulum (ER). Mapping of the mutations identified a D172G substitution mainly responsible for cadherin mislocalization. Our finding of a mutation affecting membrane receptor trafficking represents an unusual and previously unrecognized B. thuringiensis resistance mechanism.

Characterization of a Novel Member of Genus Iflavirus in Helicoverpa Armigera

Journal of Invertebrate Pathology. Mar, 2017  |  Pubmed ID: 28163012

The cotton bollworm, Helicoverpa armigera, is one of the most important agricultural pests of many economic crops worldwide. Herein, we found a novel single-strand RNA virus by RNA-Seq and Polymerase Chain Reaction (PCR) method in H. armigera named Helicoverpa armigera iflavirus (HaIV), which possessed a genome with 10,017 nucleotides in length and contained a single large open reading frame (ORF) encoding a putative polyprotein of 3021 amino acids with a predicted molecular mass of 344.16kDa and a theoretical isoelectric point (pI) of 6.45. The deduced amino acid sequence showed highest similarity (61.0%) with the protein of Lymantria dispar Iflavirus 1. Phylogenetic analysis with putative RdRp amino acid sequences indicated that the virus clustered with members of the genus Iflavirus. The virus was mainly distributed in the fat body of its host and was found to be capable of both horizontal and vertical transmission. The efficiency of perorally horizontal transmission was dose dependent (100% infection rate with a viral dose of 10(8)copies/μl) while vertical transmission efficiency was found to be relatively low (<28.57%). These results suggest that we have found a novel member of genus Iflavirus in H. armigera.

Structure and Transcription of the Helicoverpa Armigera Densovirus (HaDV2) Genome and Its Expression Strategy in LD652 Cells

Virology Journal. Feb, 2017  |  Pubmed ID: 28173863

Densoviruses (DVs) are highly pathogenic to their hosts. However, we previously reported a mutualistic DV (HaDV2). Very little was known about the characteristics of this virus, so herein we undertook a series of experiments to explore the molecular biology of HaDV2 further.

Structural Proteins of Helicoverpa Armigera Densovirus 2 Enhance Transcription of Viral Genes Through Transactivation

Archives of Virology. Feb, 2017  |  Pubmed ID: 28210815

Herein, we report the identification of putative promoters for the non-structural proteins (NS) and capsid structural proteins (VP) of Helicoverpa armigera densovirus (HaDV2) as well as a potential mechanism for how these promoters might be regulated. For the first time, we report that VP is able to transactivate the VP promoter and, to a lesser degree, the NS promoter in densoviruses. In addition to this, another promoter-like sequence designated P2, when co-transfected with the VP gene, enhanced luciferase activity by approximately 35 times compared to a control. This suggests that there are two promoters for VP in HaDV2 and that the VP of parvoviruses might play a more important role in viral transcription than previously appreciated.

TPS46, a Rice Terpene Synthase Conferring Natural Resistance to Bird Cherry-Oat Aphid, Rhopalosiphum Padi (Linnaeus)

Frontiers in Plant Science. 2017  |  Pubmed ID: 28217135

Plant terpene synthases (TPSs) are key enzymes responsible for terpene biosynthesis, and can play important roles in defense against herbivore attack. In rice, the protein sequence of TPS46 was most closely related to maize TPS10. However, unlike maize tps10, tps46 was also constitutively expressed in rice even in the absence of herbivore attack. Potential roles or constitutive emissions of specific volatiles may due to the constitutive expressions of tps46 in rice. Therefore, in the present study, RNA interference (Ri) and overexpression (Oe) rice lines were generated to investigate the potential function of TPS46 in Oryza sativa sp. japonica. Interestingly, the rice plants become more susceptible to Rhopalosiphum padi when expression of tps46 was silenced compared with Wt in greenhouse conditions. Artificial infestation bioassays further confirmed that Ri rice lines were susceptible to R. padi, whereas Oe rice lines were repellent to R. padi. Based on GC-MS and ToF-MS analysis, a total of eight volatile products catalyzed by TPS46 in rice were identified. Among them, only limonene and Eβf could be detected in all the Ri, Oe, and Wt lines, whereas other six volatiles were only found in the blend of volatiles from Oe lines. Moreover, the amount of constitutive limonene and Eβf in the Ri lines was significantly lower than in Wt lines, while the amounts of these two volatiles in the Oe line were obviously higher than in control rice. Our data suggested that the constitutive emissions of Eβf and limonene regulated by the constitutive expression of tps46 may play a crucial role in rice defense against R. padi. Consequently, tps46 could be a potential target gene to be employed for improving the resistance of plants to aphids.

Cytotoxicity and Binding Profiles of Activated Cry1Ac and Cry2Ab to Three Insect Cell Lines

Insect Science. Mar, 2017  |  Pubmed ID: 28247982

While Cry1Ac has been known to bind with larval midgut proteins cadherin, APN (amino peptidase N), ALP (alkaline phosphatase) and ABCC2 (ATP-binding cassette transporter subfamily C2), little is known about the receptors of Cry2Ab. To provide a clue to the receptors of Cry2Ab, we tested the baseline cytotoxicity of activated Cry1Ac and Cry2Ab against the midgut and fat body cell lines of Helicoverpa zea and the ovary cell line of Spodoptera frugiperda (SF9). As expected, the descending order of cytotoxicity of Cry1Ac against the three cell lines in terms of LC50 was midgut (31.0 μg/mL) > fat body (59.0 μg/mL) and SF9 cell (99.6 μg/mL). By contrast, the fat body cell line (LC50 = 7.55 μg/mL) was about 2 times more susceptible to Cry2Ab than the midgut cell line (16.0 μg/mL), whose susceptibility was not significantly greater than that of SF9 cell (27.0 μg/mL). And ligand blot showed the binding differences between Cry1Ac and Cry2Ab in the three cell lines. These results indicated that the receptors of Cry2Ab were enriched in fat body cells and thus largely different from the receptors of Cry1Ac, which were enriched in midgut cells. This article is protected by copyright. All rights reserved.

Flight Performance of Ctenoplusia Agnata (Lepidoptera: Noctuidae)

Journal of Economic Entomology. Mar, 2017  |  Pubmed ID: 28334311

Ctenoplusia agnata (Staudinger) (Lepidoptera: Noctuidae) is a highly destructive polyphagous pest of cotton, maize, soybean, and cruciferous vegetables in East Asia. The effect of various biotic and abiotic factors on the flight performance of C. agnata is crucial for a better understanding of its transregional migration. In this study, the flight performance of C. agnata moths at different ages, temperatures, and relative humidity (RH) levels, was examined by tethering individual moths to computerized flight mills for a 24-h scotophase. The results showed that 1) C. agnata had the capacity for sustained flight and the flight ability was most pronounced in 3-d-old individuals, and then their flight performance decreased significantly as the moth got older. 2) For both sexes, temperature had a significant effect on their flight performance, and the flight activity was most pronounced at 24-28 °C. 3) There was a significant effect of RH on all flight parameters of the tested moths, and the flight activity was most pronounced at RH of 60-75%. 4) For 3-d-old moths under the optimum conditions (24 °C and 75% RH) throughout the 24-h scotophase, the total flight distance reached 69.01 ± 2.13 km (females) and 62.15 ± 2.31 km (males), and the total flight duration reached 14.11 ± 0.79 h (females) and 13.08 ± 0.70 h (males), which suggests that C. agnata has a strong potential to undertake long-distance migration. These findings will be helpful for developing sound forecasting systems of this pest species.

Chimeric Antigen Receptor T Cells: a Novel Therapy for Solid Tumors

Journal of Hematology & Oncology. Mar, 2017  |  Pubmed ID: 28356156

The chimeric antigen receptor T (CAR-T) cell therapy is a newly developed adoptive antitumor treatment. Theoretically, CAR-T cells can specifically localize and eliminate tumor cells by interacting with the tumor-associated antigens (TAAs) expressing on tumor cell surface. Current studies demonstrated that various TAAs could act as target antigens for CAR-T cells, for instance, the type III variant epidermal growth factor receptor (EGFRvIII) was considered as an ideal target for its aberrant expression on the cell surface of several tumor types. CAR-T cell therapy has achieved gratifying breakthrough in hematological malignancies and promising outcome in solid tumor as showed in various clinical trials. The third generation of CAR-T demonstrates increased antitumor cytotoxicity and persistence through modification of CAR structure. In this review, we summarized the preclinical and clinical progress of CAR-T cells targeting EGFR, human epidermal growth factor receptor 2 (HER2), and mesothelin (MSLN), as well as the challenges for CAR-T cell therapy.

Expression of Lima Bean Terpene Synthases in Rice Enhances Recruitment of a Beneficial Enemy of a Major Rice Pest

Plant, Cell & Environment. Mar, 2017  |  Pubmed ID: 28370092

Volatile terpenoids play a key role in plant defence against herbivory by attracting parasitic wasps. We identified seven terpene synthase genes from lima bean, Phaseolus lunatus L. following treatment with either the elicitor alamethicin or spider mites, Tetranychus cinnabarinus. Four of which (Pltps2, Pltps3, Pltps4 and Pltps5) were upregulated, and their derived proteins are phylogenetically clustered in the TPS-g subfamily, with PlTPS3 positioned at the base of this cluster. Recombinant PlTPS3 was able to convert geranyl diphosphate and farnesyl diphosphate to linalool and (E)-nerolidol, the latter being precursor of the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). Recombinant PlTPS4 showed a different substrate specificity and produced linalool and (E)-nerolidol, as well as (E,E)-geranyllinalool from geranylgeranyl diphosphate. Transgenic rice expressing Pltps3 emitted significantly more (S)-linalool and DMNT than wild type plants, whereas transgenic rice expressing Pltps4 produced (S)-linalool, DMNT and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene. In laboratory bioassay, female parasitoid wasp Cotesia chilonis, the natural enemy of the striped rice stemborer Chilo suppressalis, were significantly attracted to the transgenic plants and their volatiles. We further confirmed this with synthetic blends mimicking natural rice volatile composition. Our study demonstrates that the transformation of rice to produce volatile terpenoids has the potential to enhance plant indirect defence through natural enemy recruitment.

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