Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Abstract Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Resumo A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-1, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-1 em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-1 como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-1 para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

Targeted innovative design of Bt Cry toxin insecticidal mimics / 生物工程学报

Bt Cry toxin is the mostly studied and widely used biological insect resistance protein, which plays a leading role in the green control of agricultural pests worldwide. However, with the wide application of its preparations and transgenic insecticidal crops, the resistance to target pests and potential ecological risks induced by the drive are increasingly prominent and attracting much attention. The researchers seek to explore new insecticidal protein materials that can simulate the insecticidal function of Bt Cry toxin. This will help to escort the sustainable and healthy production of crops, and relieve the pressure of target pests' resistance to Bt Cry toxin to a certain extent. In recent years, the author's team has proposed that Ab2β anti-idiotype antibody has the property of mimicking antigen structure and function based on the "Immune network theory" of antibody. With the help of phage display antibody library and specific antibody high-throughput screening and identification technology, Bt Cry toxin antibody was designed as the coating target antigen, and a series of Ab2β anti-idiotype antibodies (namely Bt Cry toxin insecticidal mimics) were screened from the phage antibody library. Among them, the lethality of Bt Cry toxin insecticidal mimics with the strongest activity was close to 80% of the corresponding original Bt Cry toxin, showing great promise for the targeted design of Bt Cry toxin insecticidal mimics. This paper systematically summarized the theoretical basis, technical conditions, research status, and discussed the development trend of relevant technologies and how to promote the application of existing achievements, aiming to facilitate the research and development of green insect-resistant materials.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-¹ combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-¹ level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-¹ to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-¹, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-¹ em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-¹ como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-¹ para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton

Abstract Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance ( 1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Resumo A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-1, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-1 em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-1 como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-1 para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração ( 1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

A 6-year field monitoring of fall armyworm, Spodoptera frugiperda, in transgenic Bt maize in Brazil

ABSTRACT A 6-year field monitoring study was designed to compare the presence of fall armyworm Spodoptera frugiperda (J. E. Smith), leaf injury, its parasitoids, 100 seed weight, and seed yield of Bt maize hybrids approved for commercialization in the Brazilian market. Field trials were planted in two municipalities, divided into two periods, from 2011 to 2014 (period I), and due to the approval of different Bt hybrids, from 2015 to 2016 (period II). Treatments were Bt, non-Bt maize and all non-Bt maize sprayed with insecticide methomyl. Six Bt-maize hybrids were planted, expressing the following proteins: Cry1Ab, Cry1F, Cry1A.105 (Cry1Ab +Cry1Ac +Cry1F), Cry2Ab2a and Vip3. In 2015 and 2016 Bt Powercore was planted due to its commercial approval. The number of S. frugiperda larvae for most Bt hybrids was lower than on non-Bt hybrids, except for Fórmula TL and Herculex in period II, in both municipalities. In period I, Bt hybrids VT, VT 2 and Viptera showed almost no injury on their leaves, despite the high number of fall armyworm larvae found in plants. Different parasitoids emerged from larvae from Bt and non Bt hybrids: Archytas sp., Campoletis sp., Chelonus sp., Eiphosoma sp. and Ophion luteus. The average 100 seed weight was higher in municipality from Sete Lagoas for all Bt hybrids, non-Bt hybrids and non-Bt hybrids sprayed with methomyl, when compared to Nova Porteirinha during period I. Fórmula TL, non-Bt Fórmula TL sprayed with methomyl, and non-Bt Fórmula TL showed the lowest seed yield in both municipalities and periods.

Resource use efficiency of cotton in improved vs conventional planting geometry with exogenous application of bio-stimulant and synthetic growth retardant / Eficiência do uso de recursos do algodão na geometria de plantio melhorada vs convencional com aplicação exógena de bioestimulante e retardante de crescimento sintético

Abstract Plant growth regulators and improved planting density are the innovative techniques in the establishment of more productive cotton crop. A field study was planned to assess the role of growth regulators in the resource utilization efficiency of cotton cultivars under different row spacing at Agronomic Research Area, Bahauddin Zakariya University Multan and Usmania Agricultural Farm, Shujaabad during Kharif 2012. The study was comprised of cotton cultivars viz. CIM-573 and CIM-598, cultivated under conventional (75 cm), medium (50 cm) and improved ultra-narrow row spacing (25 cm) and foliar spray of growth regulators viz. moringa leaf extract (MLE) and mepiquat chloride (MC), either alone or in combination, distilled water as a control. The application of MLE alone and in combination (MLE + MC) showed the promoting effect on crop growth rate, net assimilation rate, leaf area index, leaf area duration, sympodial branches and number of bolls leading to higher seed cotton yield of both cotton cultivars grown under conventional row spacing. While application of MC averts the plant growth without considerably improving the productivity. MLE being rich source of growth promoting hormone and nutrients showed its potential to a far greater extent under conventional row spacing in efficient utilization of available resources compared to MC and distilled water.

Resumo Reguladores de crescimento de plantas e melhor densidade de plantio são técnicas inovadoras no estabelecimento de culturas mais produtivas de algodão. Um estudo de campo foi planejado com o objetivo de avaliar o papel dos reguladores de crescimento na eficiência de uso de recursos de cultivares de algodão sob diferentes espaçamentos na Área de Pesquisa Agronômica, da Universidade Multil de Bahauddin Zakariya, e na Fazenda Agrícola da Usmania, Shujabad, durante o Kharif 2012. O estudo foi composto de cultivares de algodão CIM-573 e CIM-598, cultivados em espaçamento de linhas convencional (75 cm), médio (50 cm) e superestreito (25 cm) e de pulverização foliar de reguladores de crescimento, a saber, extrato de folhas de moringa (MLE) e cloreto de mepiquat (MC), isoladamente ou em combinação, e água destilada como controle. A aplicação de MLE isoladamente e em combinação (MLE + MC) mostrou efeito promotor na taxa de crescimento da cultura, taxa de assimilação líquida, índice de área foliar, duração de área foliar, ramos simpodiais e número de cápsulas levando à maior produção de algodão nas cultivares com espaçamento de linha convencional. Em contrapartida, a aplicação de MC evitou o crescimento da planta sem melhorar consideravelmente a produtividade. O MLE, por ser uma rica fonte de hormônio promotor de crescimento e nutrientes, mostrou seu potencial em uma extensão muito maior sob o espaçamento convencional entre as linhas no uso eficiente dos recursos disponíveis em comparação com o MC e a água destilada.

Research progress of cyclic peptides derived from phage display technology / 药学学报

Cyclic peptide drugs have gradually become an emerging research direction due to their some favorable properties such as high-efficiency binding affinity, high selectivity, lower toxicity, and stable metabolism. In recent years, the number of cyclic peptide drugs under clinical research has continued to increase. Unlike the previous cyclic peptide drugs, which were mostly derived from natural products and their derivatives, these cyclic peptide drugs are designed by genetically encoded display technologies which are based on rational design and in vitro evolution (such as BT1718, PTG-300, POL6326, etc). Among them, phage display technology has some advantages such as mature research system, low cost, and simpler operation that make it well recognized and praised by the majority of researchers in this field. Here, we reviewed the recent progress of applying phage display technology to explore diverse cyclic peptide libraries, which, we believe, will contribute more valuable candidate cyclic peptide drugs in clinical research.

Evidence of Food Safety in Relation to Cry Proteins in Genetically Modified Foods / Evidencias sobre seguridad alimentaria con relación a Cry proteínas en alimentos transgénicos

Abstract Developments in applying biotechnology to crops have generated strong ethical and social debates about its use. This study was aimed at reviewing epidemiological evidence regarding the consumption of genetically modified foods and the possible effects on human health, particularly certain insect-resistant crops in which isolated Bacillum thurigiensis Cry protein has been introduced. An in-depth review of databases was conducted for 2007-2019. Articles not referring to human health were excluded. In total, 1,350 were obtained and 118 were reviewed. As a result, it can be concluded that most studies have focused on chemical composition and in vitro or laboratory animal trials. Furthermore, the guiding principle of substantial equivalency, generally used today to evaluate potential health effects, should not replace rigorously evaluating products with nutritional, immunological, and toxicological trials. Lastly, this review demonstrates a lack of epidemiological evidence, and therefore, the safety of these foods cannot be conclusively determined based on evidence.

Resumen El desarrollo de la biotecnología aplicada a los cultivos ha generado fuertes debates éticos y sociales sobre su uso. El presente estudio tuvo por objetivo revisar las evidencias epidemiológicas existentes relacionando el consumo de alimentos genéticamente modificados, en particular aquellos provenientes de cultivos con resistencia a algunos insectos plagas en los que se han introducido proteínas Cry aisladas de Bacillum thurigiensis con probables daños o trastornos en la salud de las personas. Se realizó una revisión en profundidad en el periodo 2007 a 2019, en bases de datos. Se excluyeron aquellos artículos que no hacían referencia a salud humana. Se obtuvieron 1 350 y finalmente se revisaron 118. La revisión permitió concluir que la mayoría de los estudios existentes se centran en información respecto a la composición química y ensayos in vitro o en laboratorio con animales. Igualmente, que el principio rector de equivalencia sustancial hoy utilizado en forma generalizada para la evaluación de potenciales efectos en salud, no debería sustituir la necesidad de una evaluación rigurosa de los productos incluyendo ensayos nutricionales, inmunológicos y toxicológicos. Por último se comprueba también que la evidencia epidemiológica incluida es insuficiente por lo que lo que no es posible concluir a partir de ella, sobre la inocuidad de estos alimentos.

First record of Peridroma saucia Hübner (Lepidoptera: Noctuidae) in transgenic soybeans

ABSTRACT The widespread cultivation of transgenic soybeans has caused significant changes in the spectrum of Lepidoptera larvae, both in the number of species as well as on their densities in the field. Transgenic crops producing Bacillus thuringiensis (Bt) insecticidal proteins have successfully reduced the incidence of the most common caterpillars infesting soybeans, namely Anticarsia gemmatalis (Lepidoptera: Erebidae) and Chrysodeixis includens (Lepidoptera: Noctuidae). However, lepidopteran species not previously recorded on the crop have been recently found, and are of concern due to the possibility of adaptation to the genetically modified cultivars. The occurrence of Peridroma saucia Hübner (Lepidoptera: Noctuidae) is described for the first time in Brazil feeding on genetically modified soybean cultivars.

First record of Cotesia scotti () (Hymenoptera: Braconidae: Microgastrinae) comb. nov. parasitising Spodoptera cosmioides (Walk, 1858) and Spodoptera eridania (Stoll, 1782) (Lepidoptera: Noctuidae) in Brazil

ABSTRACT This is the first report of Cotesia scotti (Valerio and Whitfield) comb. nov. in Brazil, attacking larvae of the black armyworm, Spodoptera cosmioides, and the southern armyworm, S. eridania. The moth larvae were found respectively, infesting a protected cropping of organic tomato in Hidrolândia, Goiás, Brazil, and a transgenic soybean crop in São José dos Pinhais, Paraná, Brazil. Biological, molecular and morphological characters were used to confirm the identity of the specimens. Parasitoid identification presented a challenge since the species has most diagnostic characters of the genus Cotesia Cameron, but few in the poorly defined genus Parapanteles Ashmead. Based on morphological and molecular evidence, we transfer Parapanteles scotti to the genus Cotesia. The new combination is discussed by comparison with morphologically similar species and available molecular data.

The antitumor activity of STAT3 target inhibitor Bt354 andmolecular mechanism of anti-prostate cancer / 药学学报

Signal transducer and activator of transcription 3 (STAT3) was found in an abnormal constitutively active status in certain cancer tissues, and under these circumstances the interruption of STAT3 signaling pathway was proposed with the potential anti-cancer efficacy. In this study, our previous reported STAT3 inhibitor Bt354 can inhibit tumor growth in DU145 xenograft mice without affecting body weight. In groups treated with Bt354, the inhibition rate of tumor weight was 58.8%, 62.7% and 73.5% in 10, 20, 40 mg·kg-1 group, respectively. Particularly, the number of Ki 67 positive cells in the tumor sections was significantly decreased in Bt354 groups. Furthermore, Bt354 inhibited the nuclear translocation of STAT3 and consequently induced cell growth inhibition, apoptosis in DU145 cells. These findings suggest that Bt354 may be a potent anticancer agent for STAT3 activated prostate cancer cells. Procedures for animal study were performed with approval of the Animal Care and Use Committee of the Chinese Academy of Medical Sciences and Peking Union Medical College.

AutoBT: A new Paradigm in Periodontal regeneration

Background: Autogenous bone is an ideal material for the reconstruction of hard tissue defects, because it promotes osteogenesis, osteo-induction and osteo-conduction. The use of AutoBT, a novel bone grafting material produced from autogenous teeth, resulted in excellent bone healing based on an analysis of its inorganic components, surface structure and histologic evidence of the healing process.Materials & Methods: Ten sites were included using the following inclusion criteria. Inclusion criteria: One or more sites showing intra-bony defect with probing pocket depth (PPD) ≥ 5mm, clinical attachment loss of ≥ 3mm, 2 or 3 wall intra-bony defects with radiographic defect of size ≥ 3 mm & in same patient mobile teeth indicated for extraction. In test group, among 5 sites, regenerative treatment was performed using tooth as autograft along with chorion membrane & in control group, 5 sites were treated with demineralized freeze-dried bone allograft (DFDBA) with chorion membrane. Clinical parameters such PPD, CAL were evaluated at baseline, 3 & 6 months & radiographic parameters at baseline & after 6 months of treatment.Results: The patients treated by tooth as autoBT material with chorion membrane showed non- significant results to DFDBA with chorion membrane in intra-bony defects in all the clinical parameters. So AutoBT can be used as a useful alternative to DFDBA in periodontal regenerative therapy for intra-bony defects.

Risk Assessment of Insect-resistant and Herbicide Tolerant Genetically Modified Maize Bt11 for Cultivation, Import, Processing, Food and Feed Uses under Directive 2001/18/EC and Regulation (EC) Opinion

The Norwegian Environment Agency (NEA) and the Norwegian Food Safety Authority (NFSA) requested the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion of potential risks to biodiversity and agriculture in Norway associated with import of seeds for sowing and cultivation of insect-resistant and herbicide tolerant genetically modified maize Bt11 under Directive 2001/18/EC (Notification C/F/96.05.10). The notification is still pending for authorisation in the European Union. VKM is also requested to assess the applicant´s post-market environmental monitoring plan, and the management measures suggested in the draft implementing decision of the European Commission. As the scope of the notification does not cover food and feed uses of maize Bt11, VKM was not asked for a health risk assessment of maize Bt11. However, VKM has decided to update a previous safety evaluation of the food and feed uses of maize Bt11 and derived products (VKM, 2014). VKM appointed a working group consisting of members from the Panel on Genetically Modified Organisms, the Panel on Alien Organisms and trade in Endangered Species (CITES) and the VKM staff to answer the requests. The Panel on Genetically Modified Organisms assessed and approved the final report. The genetically modified maize Bt11 was developed to provide protection against certain lepidopteran target pests, such as the European corn borer (ECB, Ostrinia nubilalis), and some species belonging to the genus Sesamia . The insect resistence is achieved by the expression of a truncated form of a Cry1Ab protein encoded by a modified cry1Ab gene derived from the soil microorganism Bacillus thuringiensis subsp kurstaki HD-1. Maize Bt11 also expresses the phosphinothricin - N - ace tyltransferase (pat) gene, derived from the soil microorganism Streptomyces viridochromogenes strain Tu494, which encodes the enzyme: phosphinothricin acetyl transferase (PAT). PAT protein confers tolerance to the herbicidal active substance glufosinate-ammonium. The PAT protein expressed in Bt11 was used as a selectable marker to facilitate the selection process of transformed plant cells and is not intended for weed management purposes. Since the scope of the notification C/F/96.05.10 does not cover the use of glufosinate-ammonium-containing herbicides on maize Bt11, potential effects due to the use of such herbicides on maize Bt11 are not considered by VKM. In delivering its scientific opinion, VKM considered relevant peer-reviewed scientific publications and information provided by the applicant in the notification C/F/96.05.10, the renewal application EFSA/GMO/RX/Bt11, and scientific opinions and comments from EFSA and other EU-member states. VKM has evaluated maize Bt11 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010a), selection of comparators for the risk assessment of GM plants (EFSA, 2011b) and for the post-market environmental monitoring of GM plants (EFSA, 2011c). The scientific risk assessment of maize Bt11 includes molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity. An evaluation of unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes, the post-market environmental monitoring plan and coexistence measures at the farm level has also been undertaken. It is emphasised that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM. Molecular Characterization: Appropriate analyses of the integration site, inserted DNA sequence, flanking regions, and bioinformatics have been performed. The molecular characterisation reported by the applicant shows that the DNA-fragment containing the cry1Ab and pat genes, is integrated as a single copy at a single locus in the nuclear genome of maize Bt11 and that it is stably inherited as a dominant trait. VKM considers the molecular characterisation of maize Bt11 satisfactory. Comparative Assessment: Comparative analyses of data from field trials located at representative sites and environments in North America and Europe indicates that maize Bt11 is compositionally equivalent to its conventional counterpart, with the exception of the herbicide tolerance and insect resistance traits, conferred by the expression of the PAT and Cry1Ab proteins. However, data on the amino acid tryptophan, is only given in one out of six studies. Based on current knowledge, VKM concludes that maize Bt11 is compositionally equivalent to conventional maize. The data provided by the applicant are not sufficient to show that Bt11 maize is phenotypically and agronomically equivalent to conventional near-isogenic maize lines. The agronomic assessment data are provided from one growing season in the North America and one growing season in France. This is not considered to be sufficient for representative testing of agricultural environments. Food and Feed Risk Assessment: Whole food feeding studies have not indicated any adverse health effects of maize Bt11. These studies further support that maize Bt11 is nutritionally equivalent to conventional maize. The Cry1Ab and PAT proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. Some studies have however indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM concludes that maize Bt11 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry1Ab and PAT proteins will introduce a toxic or allergenic potential in food or feed based on maize Bt11 compared to conventional maize. Environmental Risk Assessment: Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivated maize with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. Since maize Bt11 has no altered agronomic and phenotypic characteristics, except for the specific target insect resistance and herbicide tolerance, the likelihood of unintended environmental effects as a consequence of spread of genes from maize Bt11 is considered to be extremely low. There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. Published scientific studies showed that the likelihood of negative effects of Cry1Ab protein on non-target arthropods that live on or in the vicinity of maize plants is low. In Norway, the maize cultivation is marginal. The total crop area of forage maize is estimated to 2000-2800 decares, equivalent to less than 0.1% of the areas with cereal crops. The area of individual fields is limited by the topography such that the quantity of maize pollen produced under flowering is also limited. The potential exposure of Cry1Ab-containing maize pollen on non-target lepidopteran species in Norway is therefore negligible. Cultivation of maize Bt11 is not considered to represent a threat to the prevalence of red-listed species in Norway. Exposure of nontarget organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Cry proteins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. VKM concludes that, although the data on the fate of the Cry1Ab protein and its potential interactions in soil are limited, the relevant scientific publications analysing the Cry1Ab protein, together with the relatively broad knowledge about the environmental fate of other Cry1 proteins, do not indicate significant direct effects on the soil environment. Despite limited number of studies, most studies conclude that effects on soil microorganisms and microbial communities are transient and minor compared to effects caused by agronomic and environmental factors. However, data are only available from short-term experiments and predictions of potential long-term effects are difficult to deduce. Coexistence: VKM concludes that separation distances of 200 meters most likely will ensure coexistence between genetically modified maize and conventional and organic maize varieties in Norway. Overall Conclusion: Based on current knowledge, VKM concludes that maize Bt11 is nutritionally equivalent t

Apoptotic effects of polyporusterone A on estrogen receptor negative breast cancer cells via regulating Bcl-2/Bax pathway / 中草药

Objective To explore the apoptotic effect of polyporusterone A on estrogen receptor (ER)-positive and ER-negative human breast cancer cells and the possible mechanism. Methods Three human breast cancer cell lines (MDA-MB-453, MCF-7, and BT474) were chosen in this study. MTT assay was performed to measure the relative cell viabilities. Flow cytometry was used to analyze the apoptosis and cell cycle. Western blot analysis was used to determine the expression levels of Bcl-2 family proteins. Results Cell proliferation of ER-negative human breast cancer cells was significantly inhibited by polyporusterone A in a dose-and time-dependent manner, while no significant effect was observed on the proliferation of strogen receptor (ER)-positive human breast cancer MCF-7 and BT474 cells. Moreover, polyporusterone A could reversibly arrest the MDA-MB-253 cells in G1 or G2/M phase. Flow cytometry results showed that 50 μmol/L polyporusterone A induced MDA-MB-253 cells apoptosis after treatment for 24 h, while no apoptosis occurred in MCF-7 and BT474 cell lines. Western blot results showed that 50 μmol/L polyporusterone A up-regulated the protein expression of Bad and Bax, but down-regulated the expression of Bcl-2 and Bcl-w protein. Conclusion Polyporusterone A can inhibit the proliferation of ER-negative breast cancer cells and promote apoptosis, which may be associated with the regulation of the expression of Bcl-2 family proteins.

El trabajo: concepciones de jóvenes que laboran en Call Centers en una ciudad intermedia colombiana / Work: Conceptions of young people working in call centers in a middle-sized Colombian city / Trabalho: Concepções de jovens que trabalham em call centers em uma cidade de tamanho médio colombiana

Las organizaciones actuales han desarrollado nuevos modelos para almacenar, transferir y transformar la información; esta era se caracteriza por su alta demanda de las nuevas tecnologías y de las características del individuo trabajador, lo que ha favorecido el desarrollo de la industria Business Process Outsourcing (BPO1), logrando atraer a nuevos trabajadores y trabajadoras jóvenes. Sin embargo, aunque se han mejorado las condiciones de empleabilidad, pareciera que su dinámica cambia tangencialmente las consideraciones sobre el trabajo. En tal sentido, en el presente artículo intento comprender los imaginarios y subjetividades del concepto de "trabajo", asumido por los sujetos jóvenes trabajadores en este tipo de organizaciones de la ciudad de Manizales. Metodológicamente, el estudio lo llevé a cabo desde una perspectiva hermenéutica mediante la aplicación de entrevistas a profundidad a las personas jóvenes, para conocer su postura y comprender la relación ser humano-trabajo. Presento los resultados desde los imaginarios, representaciones, sentido y concepción de los jóvenes y las jóvenes frente al trabajo y sus percepciones de calidad de vida, lo cual en términos generales refleja su adaptación y aceptación de las condiciones actuales de flexibilidad laboral en el país -particularmente en la región-, y la vulnerabilidad en que se encuentran. Como futuras líneas de investigación y desde algunos vacíos evidenciados, planteo la necesidad de abordar el estudio desde otras perspectivas metodológicas que permitan dar explicaciones acerca de la aceptación acrítica de las gentes jóvenes sobre las características del trabajo actual y su aporte a la construcción de nuevas prácticas laborales, al igual que indagar sobre las nuevas identidades de los individuos jóvenes.

Today's organizations have developed new models to store, transfer and transform information, which is characterized by a high demand for new technologies and related skills in workers, which has favored the development of the Business Process Outsourcing (BPO) industry. These jobs attract new and young workers, and although there are improved employment conditions, it seems that the dynamic of this industry changes how people see the notion of work. In this regard, some youth perceptions are presented against the concept of work, the sense that this population gives to their work and the conditions they perceive. Methodologically, the research was conducted used a hermeneutical perspective by conducting in-depth interviews with young people to identify their perceptions regarding work and to understand the human being-work relationship.

As organizações de hoje tém desenvolvido novos modelos para armazenar, transferir e transformar a informação, esta nova era é caracterizada pela alta demanda por novas tecnologias e características do trabalhador, o que tem favorecido o desenvolvimento de indústrias de Business Process Outsourcing (BPO), conseguindo atrair novos jovens trabalhadores, no entanto, mesmo que as condições de empregabilidade tenham melhorado, parece que sua dinã¢mica altera tangencilmente as considerações deste trabalho .Neste sentido o presente artigo tenta compreender os imaginários e subjetividades do conceito de trabalho algumas percepções dos jovens sobre o trabalho assumido por jovens que trabalham com este tipo de organizações na cidade de Manizales. Metodologicamente, o estudo foi realizado a partir de uma perspectiva hermenéutica através da aplicação de entrevistas em profundidade com os jovens para encontrar a sua posição e compreender relação homem-trabalho. Os resultados apresentam-se desde os imaginários, representações, sentido e concepção dos jovens frente ao trabalho e suas percepções de qualidade de vida, a qual em termos gerais refletem sua adaptação e aceitação laboral no país, região e vulnerabilidade em que se encontram. Como futuras linhas de investigação e alguns espaços vazios evidenciados, se plantea a necessidade de abordar o estudo desde outras perspectivas metodológicas que permitam dar explicações acerca da aceitação crítica dos jovens sobre as características do trabalho atual e seu aporte a construção de novas práticas laborais semelhante as novas identidades dos jovens.

Phytophagous mites on genetically modified maize with Bacillus thuringiensis genes / Ácaros fitófagos em milho geneticamente modificado com genes de Bacillus thuringiensis

ABSTRACT: The study of non-target organisms such as mites, can provide information about the possible effects of transgenic maize cultivars on the arthropod community. This study aimed to evaluate the abundance of phytophagous mites in the area of Bt maize and evaluate the instantaneous population growth rate, and food preference of phytophagous mites on Bt maize. We registered the species occurrence and the number of adult mite individuals on four maize cultivars, one non-Bt 30F35, and three containing the proteins Cry1Ab (30F35 Yg), Cry1F (30F35 Hx) and Vip3a (Impacto Viptera). Cry proteins disrupt the midgut epithelium of insect pests. The food preference and instantaneous population growth rate (ri) were evaluated using the two-spotted spider mite Tetranychus urticae as model. The species Catarhinus tricholaenae and Aceria zeala were recorded. The Bt maize cultivars did not significantly affect the average number of C. tricholaenae and A. zeala mites compared to conventional cultivar. The population growth rates of T. urticae were similar for the different maize cultivars. T. urticae showed no preference between the leaf of Bt or conventional cultivars. Bt maize did not affect the abundance of species in phytophagous mite.

RESUMO: O estudo de organismos não-alvo, tais como ácaros, pode fornecer informações sobre os possíveis efeitos de cultivares de milho transgênico na comunidade de artrópodos. Este estudo teve como objetivo avaliar a abundância de ácaros fitófagos na área de milho Bt e avaliar a taxa de crescimento populacional instantânea e preferência alimentar de ácaros fitófagos em milho Bt. Registramos a ocorrência de espécies e número de indivíduos adultos de ácaros em quatro cultivares de milho, um não-Bt 30F35, e três contendo a proteína Cry1Ab (30F35 Yg), Cry1F (30F35 Hx) e VIP3A (Impacto Viptera). A preferência alimentar e taxa instantânea de crescimento populacional foram avaliadas utilizando fêmeas adultas do ácaro-rajado Tetranychus urticae. As espécies Catarhinus tricholaenae e Aceria zeala foram registradas. As cultivares de milho Bt não afetaram significativamente o número médio de ácaros C. tricholaenae e A. zeala em relação ao cultivar convencional. As taxas de crescimento populacional de T. urticae foram similares para os diferentes cultivares. T. urticae não mostrou preferência entre a folha de milho Bt ou cultivares convencionais. Sugere-se, então, que o milho Bt não afeta a abundância de espécies, preferência alimentar e taxa de crescimento de ácaros fitófagos.

Resource use efficiency of cotton in improved vs conventional planting geometry with exogenous application of bio-stimulant and synthetic growth retardant

Abstract Plant growth regulators and improved planting density are the innovative techniques in the establishment of more productive cotton crop. A field study was planned to assess the role of growth regulators in the resource utilization efficiency of cotton cultivars under different row spacing at Agronomic Research Area, Bahauddin Zakariya University Multan and Usmania Agricultural Farm, Shujaabad during Kharif 2012. The study was comprised of cotton cultivars viz. CIM-573 and CIM-598, cultivated under conventional (75 cm), medium (50 cm) and improved ultra-narrow row spacing (25 cm) and foliar spray of growth regulators viz. moringa leaf extract (MLE) and mepiquat chloride (MC), either alone or in combination, distilled water as a control. The application of MLE alone and in combination (MLE + MC) showed the promoting effect on crop growth rate, net assimilation rate, leaf area index, leaf area duration, sympodial branches and number of bolls leading to higher seed cotton yield of both cotton cultivars grown under conventional row spacing. While application of MC averts the plant growth without considerably improving the productivity. MLE being rich source of growth promoting hormone and nutrients showed its potential to a far greater extent under conventional row spacing in efficient utilization of available resources compared to MC and distilled water.

Resumo Reguladores de crescimento de plantas e melhor densidade de plantio são técnicas inovadoras no estabelecimento de culturas mais produtivas de algodão. Um estudo de campo foi planejado com o objetivo de avaliar o papel dos reguladores de crescimento na eficiência de uso de recursos de cultivares de algodão sob diferentes espaçamentos na Área de Pesquisa Agronômica, da Universidade Multil de Bahauddin Zakariya, e na Fazenda Agrícola da Usmania, Shujabad, durante o Kharif 2012. O estudo foi composto de cultivares de algodão CIM-573 e CIM-598, cultivados em espaçamento de linhas convencional (75 cm), médio (50 cm) e superestreito (25 cm) e de pulverização foliar de reguladores de crescimento, a saber, extrato de folhas de moringa (MLE) e cloreto de mepiquat (MC), isoladamente ou em combinação, e água destilada como controle. A aplicação de MLE isoladamente e em combinação (MLE + MC) mostrou efeito promotor na taxa de crescimento da cultura, taxa de assimilação líquida, índice de área foliar, duração de área foliar, ramos simpodiais e número de cápsulas levando à maior produção de algodão nas cultivares com espaçamento de linha convencional. Em contrapartida, a aplicação de MC evitou o crescimento da planta sem melhorar consideravelmente a produtividade. O MLE, por ser uma rica fonte de hormônio promotor de crescimento e nutrientes, mostrou seu potencial em uma extensão muito maior sob o espaçamento convencional entre as linhas no uso eficiente dos recursos disponíveis em comparação com o MC e a água destilada.

Effect of NUP88 gene on proliferation and invasion biological behavior of breast cancer cell line BT-20 / 中国免疫学杂志

Objective:To observe the effect of low-expression or over-expression of NUP88 gene on the proliferation and invasion ability of breast cancer cell line BT-20.Methods: NUP88 recombinant adenovirus expression vector and NUP88 RNAi adenovirus vector were transfected into breast cancer BT-20 cells to obtain BT-20 cells over-expressing NUP88 and BT-20 cells lower-expressing NUP88 and then detected the expression of NUP88 mRNA and NUP88 protein.After that,the apoptosis of BT-20 cells was detected by flow cytometry and the invasion and metastasis of BT-20 cells were detected by Transwell invasion assay.The expression of apoptosis protein and invasion and metastasis proteins were detected by Western blot.Results: BT-20 cell with the over expression levels of NUP88 mRNA and NUP88 protein and BT-20 cell with the low expression levels of NUP88 mRNA and NUP88 protein were structured.The over-expression of NUP88 gene led to proliferation rate and the number of invasive cells were significantly higher than BT-20 cells,apoptosis cells were significantly lower than BT-20 cells(P<0.05).However,the low-expression of NUP88 gene led to proliferation rate and the number of invasive cells were significantly lower than BT-20 cells,apoptosis cells was significantly higher than BT-20 cells(P<0.05).The over-expression of NUP88 gene led to Bcl-2 and β-catenin level were significantly higher than that of BT-20 cells,and Bax and E-cadherin level were significantly lower than that of BT-20 cells(P<0.05).However,the low-expression of NUP88 gene led to Bcl-2 and β-catenin level were significantly lower than that of BT-20 cells,and Bax and E-cadherin level were significantly higher than that of BT-20 cells(P<0.05).Conclusion: NUP88 gene regulates the proliferation and invasion and migration ability of breast cancer cells by regulating the expression of Bax,Bcl-2,E-cadherin and β-catenin.It has an important significance in the target treatment of breast cancer.

Toxicity of Bacillus thuringiensis Cry2Ab and the inheritance of Cry2Ab resistance in the Pink bollworm, Pectinophora gossypiella (Saunders).

Though the insecticidal crystal (Cry) proteins produced by Bacillus thuringiensis (Bt) are effective against insect pests, evolution of resistance remains an issue of great concern. Here, we explored the ability of pink bollworm, Pectinophora gossypiella (Lepidoptera: Gelechiidae), a key pest of cotton, to evolve resistance to Cry2Ab and characterized it in terms of inheritance. Sixteen day bioassay of Bt Cry2Ab toxin against 5-day old pink bollworm larvae showed median lethal concentration (LC50) ranging 0.16-1.44 µg/g diet for five different populations collected from Srivilliputtur (Tamil Nadu), Jalgaon (Maharashtra), Bharuch (Gujarat), New Delhi (Delhi) and Sri Ganganagar (Rajasthan). Selection of pink bollworm for evolution of Cry2Ab resistance led to the maximal of 37.75-fold resistance vis-à-vis the most susceptible strain. Further studies on inheritance using above parental populations showed autosomal and semidominant nature of Cry2Ab resistance, with dominance h values of 0.69 and 0.79 for two reciprocal crosses. The inheritance of Cry2Ab resistance appeared to be governed by multiple alleles/genes. Cry2Ab resistance was associated with fitness costs in terms of prolonged larval and pupal period when resistant parent population was reared on the diet without toxin. Fitness cost in terms of larval period appeared to be inherited in F1, F2 and backcross progenies. F2 progeny also inherited these differences in larval and pupal periods. These studies clearly advocate rigorous monitoring of Cry2Ab resistance and compliance of resistance management in the pink bollworm for ensuring Bt cotton sustainability.

Expression of cry1Ab gene from a novel Bacillus thuringiensis strain SY49-1 active on pest insects

ABSTRACT In this study, the cry1Ab gene of previously characterized and Lepidoptera-, Diptera-, and Coleoptera-active Bacillus thuringiensis SY49-1 strain was cloned, expressed and individually tested on Ephestia kuehniella (Lepidoptera: Pyralidae) and Plodia interpunctella (Lepidoptera: Pyralidae) larvae. pET-cry1Ab plasmids were constructed by ligating the cry1Ab into pET28a (+) expression vector. Constructed plasmids were transferred to an Escherichia coli BL21 (DE3) strain rendered competent with CaCl2. Isopropyl β-D-1-thiogalactopyranoside was used to induce the expression of cry1Ab in E. coli BL21(DE3), and consequently, ∼130 kDa of Cry1Ab was obtained. Bioassay results indicated that recombinant Cry1Ab at a dose of 1000 µg g-1 caused 40% and 64% mortality on P. interpunctella and E. kuehniella larvae, respectively. However, the mortality rates of Bt SY49-1 strains' spore-crystal mixture at the same dose were observed to be 70% on P. interpunctella and 90% on E. kuehniella larvae. The results indicated that cry1Ab may be considered as a good candidate in transgenic crop production and as an alternative biocontrol agent in controlling stored product moths.