Establishment of maize resistance to fungal diseases by host-induced gene silencing and site-directed mutagenesis

Maize is one of the most cultivated crops in the world. A disease called anthracnose accounts for up to 80% of the loss in maize production. It is caused by the hemibiotrophic fungus Colletotrichum graminicola. Unfortunately, the disease is notoriously difficult to combat, since host resistance mechanisms are hardly available. In the present investigation, the principle of host-induced gene silencing (HIGS) was employed to protect maize plants from C. graminicola infection. HIGS is an RNA-interefence (RNAi)-based process, wherein plant-produced short interfering RNAs (siRNA) are taken up by the fungus and trigger the silencing of cognate genes of the latter. In the present study, genes encoding fungicide targets were chosen as HIGS targets, namely C. graminicola -Tubulin 2 and Succinate dehydrogenase 1. RNAi vectors were designed using appropriate regions of these target genes. Transgenic plants expressing RNAi constructs were infected with C. graminicola, whereby the plants showed quantitative resistance. In addition to the HIGS approach, a further strategy was pursued, which consisted in knocking out a susceptibility factor against C. graminicola by means of targeted mutagenesis. This factor was the 9-LIPOXYGENASE LOX3 gene from maize, for which several mutated plants were generated by expression of RNA-directed Cas9 endonuclease. Homozygous lox3 mutants were tested in C. graminicola infection assays to analyze the consequences of their mutations. Quantification of fungal biomass revealed that the lox3 mutants were significantly less colonized by C. graminicola compared to the non-mutated wild-type. Corn common smut, another important fungal disease, is caused by the biotrophic pathogen Ustilago maydis. Transcriptional data (Doehlemann et al., 2008) collected during the course of infection with U. maydis showed that, depending on the infection, several members of the LOX gene family are upregulated, one of which is LOX3. Therefore, the available lox3 mutants were tested for their response to infection with U. maydis. The quantification of the disease symptoms showed that the lox3 mutants showed a moderate resistance against U. maydis infections. Furthermore, the quantification of the biomass of U. maydis revealed that the lox3 mutants were colonized by the fungus to a lesser extent compared to the wild-type. Furthermore, infection tests were performed using lox3 mutants independently produced by transposon insertion mutagenesis. These lines showed a resistance behavior similar to that of Cas9-induced mutants, by which the anticipated role of LOX3 for the interaction of maize and U. maydis was corroborated. From the literature it is known that U. maydis suppresses the accumulation of reactive oxygen species (ROS) to establish its biotrophic mode of pathogenesis. A ROS accumulation test revealed that lox3 mutants feature increased ROS accumulationcompared to the wild-type, suggesting that the immunity of the mutants triggered by pathogen-associated molecular pattern (PAMP) led to a reduction in the severity of fungal infection. This is the first study showing that lox3 mutants show moderate resistance to U. maydis. In view of these results, it is concluded that LOX3 is a susceptibility factor for U. maydis as wel

Thin Films of Tin Sulphide for Application in Photovoltaic Solar Cells

Tin sulphide (SnS) is a promising new material for use in photovoltaic solar cells. With a direct energy band gap of about 1.3 eV, and a high optical absorption coefficient, only a few microns of SnS are needed to absorb most of the incident light. Not only is SnS made of abundant, environmentally acceptable elements, it is also amphoteric giving flexibility to device design. Structures that can be envisioned include p-type SnS (absorber layer) / n-type (window layer) heterojunction devices, buried p-n junction devices made using SnS and p-i-n structure devices where the i-layer is SnS. It is most likely that the grain boundaries in SnS can be passivated either by counter-doping the grain boundaries, or by oxidizing the grain boundaries to form wide energy bandgap n-type SnO2 within p-type SnS, as dopants or oxygen will diffuse preferentially down the grain boundaries and react first at the grain boundary surfaces. Thin film solar cell devices based on the use of SnS have now been produced with efficiencies > 2 %; these and other promising results indicate that it is most likely that devices with efficiencies > 10% will be produced in the near future. Given that tin layers are routinely coated in industry over large area substrates and that industrial sulphidization processes are also well established, the industrialization of this technology should be more straightforward than that encountered with the already commercialised cadmium telluride and copper indium gallium diselenide thin film technologies. This review discusses the chemical and physical properties of SnS, the methods of producing both bulk crystals and thin films of SnS, the literature available on studies of SnS2 based photovoltaic solar cell devices, and progress made so far in developing this exciting new material

Formulation development and In vivo evaluation of mouth dissolving films containing Palonosetron HCl

Fast dissolving drug delivery systems have gained patient acceptability and popularity in the recent times. The purpose of this work was to develop mouth dissolving oral films of Palonosetron HCl is an antiemetic drug especially used in the prevention and treatment of chemotherapy-induced nausea and vomiting . An attempt was made to prepare oral dissolving films by solvent casting method. The films were prepared by using different grades of HPMC E3, E6 and E15, maltodextrin DE6 and other polymers by solvent casting method. They were evaluated for physical characteristics such as thickness, uniformity of weight, folding endurance, drug content, surface pH, percentage elongation and tensile strength and results were found to be satisfactory. The formulations were subjected to disintegration and in-vitro drug release test.The in vitro disintegration time of the optimized formulation F13 was10sec and drug release was found to be very fast i.e. 99.52% of within 10 min when compared with innovator product i.e 80.5%. In vivo studies confirmed that their potential as an innovative dosage form to improve the bioavailability and considered to be potentially useful for the treatment of emesis where quick onset of action is desirable. DSC and FTIR data revealed that no interactions takes place between the drug and polymers used in the optimized formulation. From the above results, it can be a good alternative to conventional Palonosetron tablets in the treatment of emesis. In vitro and in vivo evaluation of the films confirmed their potential as an innovative dosage form to improve delivery and quick onset of action of Granisetron Hydrochloride. Therefore, the oral fast dissolving film is considered to be potentially useful for the treatment of emesis disease where quick onset of action is desired, also improved patient compliance

Investigation of genetic algorithm design representation for multi-objective truss optimization

The objective of this research is to develop a flexible design grammar and genetic algorithm representation to be used in a multi-objective optimization method to design efficient steel roof trusses given space dimensions and loading requirements by the user. The goal of implementing the method as a multi-objective problem is to obtain a set of near-optimal trusses for the defined unstructured problem domain, not just a single near-optimal design. The method developed was required to support the exploration of a broad range of conceptual designs before making design decisions. Therefore, a method was developed that could define numerous design variables, support techniques to locate global or near-global optimal designs, and improve the efficiency of the computational procedures implemented. This research effort was motivated by the need to consider structural designs that may be beyond the established conventions of designers in the search for cost-efficient, structurally-sound designs. An effective design grammar that is capable of generating stable trusses is defined in this research. The design grammar supports the optimization of member size, in addition to truss geometry and topology. Multi-objective genetic algorithms were used to evolve sets of Pareto-optimal trusses that had varying topology, geometry, and member sizes. The Pareto-optimal curves provided design engineers with a range of near-optimal design alternatives that showed the tradeoffs that occur in meeting the stated objectives. Designers can select their final design from this set based on their own individual weighting of the design objectives. Trials are performed using a multiobjective genetic algorithm that works with the design grammar to evolve trusses for different span lengths. In addition to evaluate the performance of the developed optimization method further, trials were performed on a benchmark truss problem domain and the results obtained were compared with results obtained by other researchers. The results of the performance evaluation trials for the proposed method, in which the sizing, shape and topology were simultaneously performed, indicated that the method was effective in evolving a variety of truss topologies compared to previous published results, which evolved from a ground structure. The diverse topologies, however, were obtained over several trials instead of being found in a Pareto-optimal set found by a single trial. In addition, the proposed method was not able to locally optimize the member section sizes. Additional trials were performed to determine the benefit of applying local optimization to the member section sizes for a given truss topology or geometry provided by the method. The results indicate that significant weight reduction could be achieved by performing local optimization to the truss designs obtained by the proposed multi-objective optimization method

Mechanisms of Action of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) in Colon Cancer

Non-steroidal anti-inflammatory drugs (NSAIDs) and their NO derivatives (NO-NSAIDs), and synthetic analogs are highly effective as anticancer agents that exhibit relatively low toxicity compared to most clinically used drugs. However, the mechanisms of action for NSAIDs and NO-NSAIDs are not well defined and this has restricted their clinical applications and applications for combined therapies. Earlier studies from our laboratory reported that specificity protein (Sp) transcription factors (Sp1, Sp3 and Sp4) are overexpressed in several types of human cancers including colon cancer and many Sp-regulated genes are pro-oncogenic and individual targets for cancer chemotherapy. Based on published results showing that NSAIDs downregulate several putative Sp-regulated genes, we hypothesized that the anticancer properties of NSAIDs may be due, in part, to downregulation of Sp transcription factors. NSAIDs including aspirin and tolfenamic acid (TA) and nitro derivatives of NSAIDs such as GT-094 have been investigated in colon cancer cells and in vivo xenograft models. Aspirin and TA induced apoptosis and decreased colon cancer cell growth and tumor growth in vivo and downregulated genes associated with cell growth, survival, and angiogenesis. Previous RNA interference studies in this laboratory have shown that many of these genes are regulated, in part, by Sp transcription factors Sp1, Sp3 and Sp4 that are overexpressed in colon and other cancer cell lines. Not surprisingly, these NSAIDs also decreased Sp1, Sp3 and Sp4 proteins and Sp-regulated gene products in colon cancer cells and this was due to caspase-dependent proteolysis of Sp1, Sp3 and Sp4 proteins. Aspirin-induced activation of caspases and degradation of Sp1, Sp3 and Sp4 was due to sequestration of zinc and could be reversed by addition of zinc sulphate, whereas TA mediated induction of caspases was independent of zinc ions and is currently being investigated. GT-094 is a novel NO chimera-containing NSAID, which also inhibited colon cancer cell proliferation and induced apoptosis; these effects were accompanied by decreased mitochondrial membrane potential (MMP) and induction of reactive oxygen species (ROS), and were reversed after cotreatment with the antioxidant glutathione. GT-094 also downregulated Sp and Sp-dependent gene products and was due to decreased expression of microRNA-27a (miR-27a) and induction of ZBTB10, an Sp transcriptional repressor that is regulated by miR-27a in colon cancer cells. Moreover, the effects of GT-094 on Sp1, Sp3, Sp4, miR-27a and ZBTB10 were also inhibited by glutathione suggesting that the anticancer activity of GT-094 in colon cancer cells is due, in part, to ROS-dependent disruption of miR-27a:ZBTB10. The importance of ROS induction in targeting Sp transcription factors was also confirmed using pro-oxidants such as ascorbic acid, hydrogen peroxide and t-butyl hydroperoxide and similar results have been observed in collaborative studies with other ROS inducers in colon cancer cells. Many cancer cell lines and tumors exhibit addiction to non-oncogenes such as Sp1, Sp3 and Sp4 for maintaining the oncogenic phenotype and future research will focus on the mechanisms of ROS-mediated targeting of Sp transcription factors which represents a novel approach for cancer chemotherapy

1 STUDI ANALISIS PEMBENTUKAN CIVIC VIRTUE DALAM RUANG LINGKUP NORMA, HUKUM DAN PERATURAN DI SMP NEGERI 1 GEMOLONG TAHUN 2009

Tantangan pendidikan dewasa ini untuk menghasilkan Sumber Daya Manusia (SDM) yang berkualitas dan tangguh semakin berat. Pendidikan tidak cukup hanya berhenti pada memberikan pengetahuan yang paling mutakhir, namun juga harus mampu membentuk dan membangun sistem keyakinan dan karakter kuat setiap peserta didik sehingga mampu mengembangkan potensi diri dan menemukan tujuan hidupnya. Pendidikan di sekolah tidak lagi cukup hanya dengan mengajar peserta didik membaca, menulis, dan berhitung kemudian lulus ujian dan nantinya mendapatkan pekerjaan yang baik. Sekolah juga harus mampu mendidik peserta didik agar dapat memutuskan apa yang benar dan salah. Sekolah juga perlu membantu menemukan tujuan hidup setiap peserta didik. Bangsa Indonesia menyadari pentingnya pendidikan untuk mengembangkan potensi SDM secara optimal untuk menjadikan insan pembangunan yang berkualitas. Hal tersebut tertuang dalam Undang-Undang Dasar 1945 (UUD 1945) pada pasal 31 ayat (1), (2), (3), (4), dan (5) yang menyatakan bahwa: 1. Setiap warga negara berhak mendapat pendidikan. 2. Setiap warga negara wajib mengikuti pendidikan dasar dan pemerintah wajib membiayainya. 3. Pemerintah mengusahakan dan menyelenggarakan suatu sistem pendidikan nasional yang meningkatkan keimanan dan ketakwaan serta akhlak mulia dalam rangka mencerdaskan kehidupan bangsa yang diatur dengan undang undang. 4. Negara memprioritaskan anggaran pendidikan sekurang-kurangnya dua puluh persen dari anggaran pendapatan dan belanja daerah untuk memenuhi kebutuhan penyelenggaraan pendidikan nasional. 5. Pemerintah memajukan ilmu pengetahuan dan teknologi dengan menjunjung tinggi nilai-nilai agama dan persatuan bangsa untuk kemajuan peradaban serta kesejahteraan umat manusia. (Anonim, 2005:23) Adanya pernyataan bahwa tujuan negara Republik Indonesia yang tercantum dalam Pembukaan UUD 1945 yaitu mencerdaskan kehidupan bangsa, maka yang terjadi adalah banyaknya tuntutan dan kebutuhan bagi bangsa ini menjadi negara yang maju dan mandiri. Untuk itu faktor yang penting adalah sumber daya manusia, karena sumber daya manusia sebagai faktor yang sangat strategis sekali. Artinya tuntutan dan kebutuhan utama adalah mengembangkan kreativitas dan inovasi, kemampuan dalam mengembangkan dan mendayagunakan teknologi, dan sekaligus kemampuan dalam mengelola dan mengembangkan pembangunan nasional dalam bidang pendidikan. Upaya untuk mewujudkan pembangunan nasional dalam bidang pendidikan perlu adanya kesadaran guru memahami tentang fungsi pendidikan, dengan harapan guru dapat menjadikan peserta didik sesuai dengan fungsi dan tujuan pendidikan nasional yang tercantum dalam Undang-Undang Nomor 20 Tahun 2003 tentang Sistem Pendidikan Nasional (UU Sisdiknas). Sehingga dengan terwujudnya pembangunan nasional dalam bidang pendidikan diharapkan dapat mencapai tujuan pendidikan nasional yang salah satunya adalah membentuk manusia Indonesia yang berkualitas yang mampu menghadapi segala tantangan di segala bidang. UU Sisdiknas pasal 3 menyatakan Pendidikan nasional berfungsi mengembangkan kemampuan dan membentuk watak serta peradaban bangsa yang bermartabat dalam rangka mencerdaskan kehidupan bangsa, bertujuan untuk berkembangnya potensi peserta didik agar menjadi manusia yang beriman dan bertakwa kepada Tuhan Yang Maha Esa, berakhlak mulia, sehat, berilmu, cakap, kreatif, mandiri, dan menjadi warga negara yang demokratis serta bertanggung jawab (Anonim, 2003:3). Upaya untuk mewujudkan fungsi dan tujuan pendidikan nasional guru dituntut untuk profesional. Seperti yang tercantum pada Undang-Undang Nomor 14 Tahun 2005 tentang Guru dan Dosen pasal 1 ayat (1) bahwa “Guru adalah pendidik profesional dengan tugas utama mendidik, mengajar, membimbing, mengarahkan, melatih, menilai, dan mengevaluasi peserta didik pada pendidikan anak usia dini jalur pendidikan formal, pendidikan dasar, dan pendidikan menengah” (Anonim, 2006:2). Hal tersebut dapat dimaknai bahwa guru sebagai tenaga profesional mempunyai peran sebagai agen dalam meningkatkan mutu pendidikan nasional

Interaction of Ihh and BMP/Noggin Signaling during Cartilage Differentiation

AbstractBone morphogenetic proteins (BMPs) have been implicated in regulating multiple stages of bone development. Recently it has been shown that constitutive activation of theBMP receptor-IAblocks chondrocyte differentiation in a similar manner as misexpression ofIndian hedgehog.In this paper we analyze the role of BMPs as possible mediators of Indian hedgehog signaling and useNogginmisexpression to gain insight into additional roles of BMPs during cartilage differentiation. We show by comparative analysis ofBMPandIhhexpression domains that the borders ofIndian hedgehogexpression in the chondrocytes are reflected in changes of the expression level of severalBMPgenes in the adjacent perichondrium. We further demonstrate that misexpression ofIndian hedgehogappears to directly upregulateBMP2andBMP4expression, independent of the differentiation state of the flanking chondrocytes. In contrast, changes inBMP5andBMP7expression in the perichondrium correspond to altered differentiation states of the flanking chondrocytes. In addition,NogginandChordin,which are both expressed in the developing cartilage elements, also change their expression pattern afterIhhmisexpression. Finally, we use retroviral misexpression ofNoggin,a potent antagonist of BMP signaling, to gain insight into additional roles of BMP signaling during cartilage differentiation. We find that BMP signaling is necessary for the growth and differentiation of the cartilage elements. In addition, this analysis revealed that the members of the BMP/Noggin signaling pathway are linked in a complex autoregulatory network

Development of a Modular Unit of a Higher Level Framework or Tool for Basic Programming Course Teaching Through E-Learning Mode

This paper reports about the development of a modular unit of a higher level framework or tool whose intended objective is the creation of animated lessons for basic programming (CS1) course in computer science discipline with visual aids. The goal of such lessons is to address the difficulties faced by the novice programmers in CS1 course.This module here after referred to as ‘type writer’allows instructors to render programmes or code snippets in a live typing manner as opposed to their sudden or en-block placement on the presentation area like a Power Point Slide; a commonly used approach in the present day eLearning.This project is planned to be executed in two stages and ‘type writer’ is the outcome of the first stage. This would be combined with another two modules that are planned to be developed in the second stage, to make the complete tool. The entire tool would be developed in Action Script 3.0 language that works on Adobe Flash Platform