Delivering Signal-Altering Bacterial Effector Proteins to Mammalian Cells Using Cell-Penetrating Peptide Technology

A major role of the mitogen activated protein kinase (MAPK) pathway in eukaryotes is to activate the bacterial pathogen defense response upon the detection of bacterial products in the environment. This defensive signaling results in the induction of inflammation, the transcription of antimicrobial peptides, the modulation of the cell cycle and cell survival. Some Gram-negative bacteria have evolved needle-like structures called Type III Secretion Systems (T3SS) that secrete signal-altering molecules into the host cell to interrupt signaling pathways that would otherwise lead to the elimination of the bacterial infection. These signal-altering molecules are known as bacterial effector proteins (BEPs). Bacterial effectors YopJ and VopA have been shown to interfere with specific signaling molecules in the MAPK pathway; effectively inducing apoptosis in mammalian intestinal endothelial cells. In this study, we deliver these proteins to colon cancer cells to artificially induce cell death, using a novel cell-penetrating peptide (CPP) delivery system called TAT-CaM. Here, we show that the TAT-CaM system is capable of delivering YopJ into mammalian cells and that YopJ is capable of inducing cell death once delivered. Although we encountered issues with reproducibility, we believe that TAT-CaM-YopJ could be effective in inducing cell death in cancer cells in a reproducible manner after experimental adjustments

Idiopathic scoliosis: aetiology, natural history and treatment

EPIDEMIOLOGY: 1 | School Screening for Scoliosis: cohort study of clinical course. 2 | Assessment of Scoliosis is Children - low dose radiographic technique. 3 | Scoliosis in the Community. 4 | School Screening and Pelvic Tilt Scoliois. 5 | Screening for Scoliosis. 6 | Stature and Idiopathic Scoliosis. A Prospective Study. 7 | Scoliosis in the Community. 8 | Screening for Scoliosis: the problem of arm length. 9 | Idiopathic Scoliosis: The Leeds Epidemiology Survey. 10 | The First Year's Follow-up of the MRC Scoliosis Screening Programme.PATHOGENESIS: 11 | The Pathogenesis of Idiopathic Scoliosis - bi-planar spinal asymmetry. 12 | The Pathogenesis of Idiopathic Scoliosis. 13 | Idiopathic Scoliosis in Three Dimensions. 14 | Combined Idiopathic Kyphosis and Scoliosis. An Analysis of the Lateral Spinal Curvatures Associated with Scheuermann's disease. 15 | Aetiology of Idiopathic Spinal Deformities. 16 | The Pathogenesis of Idiopathic Scoliosis. 17 | Scoliosis: How Big Are You? 18 | The Anatomy of Spinal Deformity: A biomechanical Analysis. 19 | Vertebral Shape in the Median Sagittal Plane in Idiopathic Thoracic Scoliosis. A study of true lateral radiographs on 150 patients. 20 | Spinal Growth. 21 | The Aetiology of Spinal Deformities.EXPERIMENTAL SCOLIOSIS: 22 | The Experimental Basis of Idiopathic Scoliosis. 23 | Experimental Structural Scoliosis.CLINICAL APPLICATION: 24 | Conservative Treatment for Idiopathic Scoliosis. 25 | The Surgical Management of Idiopathic Thoracic Scoliosis. 26 | Spinal Deformities in Children. 27 | Operative Surgery for Spinal Deformity. 28 | Idiopathic Scoliosis: Foundation for Physiological Treatment. 29 | Surgical Treatment of Late-onset Idiopathic Thoracic Scoliosis - The Leeds Procedure.MISCELLANEOUS: 30 | The Kyphotic Spine in Myelomeningocele. 31 | Vertebral Body Resection for Spinal Deformity. 32 | Congenital Lumbar Kyphosis in Myelomeningocele - vertebral body resection and posterior fusion. 33 | Cotrel Traction, Exercises, Casting in the Treatment of Idiopathic Scoliosis - a Prospective Controlled Clinical Trial. 34 | Two-Stage Corrective Surgery for Congenital 35 | Deformities of the Spine. Management of Spinal Deformities

Method for Obtaining Antifungal and Herbicidal Compounds that Target the First Committed Step in Shingolipid Long-Chain Base Biosynthesis

The invention provides the LCB1 and LCB2 genes of the yeast Saccharomyces cerevisiae that encode subunits of the enzyme serine palmitoyltransferase (SPT), the first enzyme leading to synthesis of the long-base component of the sphingolipids. The present specification describes the isolation of the LCB1 and LCB2 genes. The invention further relates to methods of using these genes to either inhibit SPT activity or to inhibit synthesis of the enzyme. Furthermore, the invention relates to methods for constructing strains of S. cerevisiae or other organisms that can be used to select and to test for compounds that either inhibit SPT activity or to inhibit synthesis of the enzyme

A Geographical Location Model for Targeted Implementation of Lure-and-Kill Strategies Against Disease-Transmitting Mosquitoes in Rural Areas

Outdoor devices for luring and killing disease-transmitting mosquitoes have been proposed as potential com- plementary interventions alongside existing intra-domiciliary methods namely insecticide treated nets and house spraying with residual insecticides. To enhance effectiveness of such outdoor interventions, it is essential to optimally locate them in such a way that they target most of the outdoor mosquitoes. Using odour-baited lure and kill stations (OBS) as an example, we describe a map model derived from: 1) com-munity participatory mapping conducted to identify mosquito breeding habitats, 2) entomological field studies conducted to estimate outdoor mosquito densities and to determine safe distances of the OBS from human dwellings, and 3) field surveys conducted to map households, roads, outdoor human aggregations and landmarks. The resulting data were combined in a Ge- ographical Information Systems (GIS) environment and analysed to determine optimal locations for the OBS. Separately, a GIS-interpolated map produced by asking community members to rank different zones of the study area and show where they expected to find most mosquitoes, was visually compared to another map interpolated from the entomological survey of outdoor mosquito densities. An easy-to-interpret suitability map showing optimal sites for placing OBS was produced, which clearly depicted areas least suitable and areas most suitable for locating the devices. Comparative visual interpretation of maps derived from interpolating the community knowledge and entomological data revealed major similarities between the two maps. Using distribution patterns of human and mosquito populations as well as characteristics of candidate outdoor interventions, it is possible to readily determine suitable areas for targeted positioning of the interventions, thus improve effectiveness. This study also highlights possibilities of relying on community knowledge to approximate areas where mosquitoes are most abundant and where to locate outdoor complementary interventions such as odour-baited lure and kill stations for controlling disease-transmitting mosquitoes.\u

Social Networking in Academic Libraries: The Possibilities and the Concerns

The goal of this article is to examine the use of the major social networking tools in academic libraries in the United States. Since college students are heavy users of social networking, such efforts provide academic libraries with outreach possibilities to students who do not use the physical library. The paper also examines the concerns about their use both from students and within the academic library

Fungal IPC Synthase Assay

The presently-disclosed IPC synthase-inhibitor assays comprise the steps of: (1) expression of the IPC1 gene in a cell; (2) introducing labeled starting substrates for ceramide conversion as well as potential inhibitor(s) of such conversion to the expressed gene product in an environment which allows time and conditions for conversion, and (3) identifying those potential inhibitors which actually inhibit conversion. The present invention also provides methods to determine the ability of a test compound to inhibit fungal growth, comprising the steps of (1) presenting active inositolphosophotidylceramide synthase in a manner such that synthesis of inositolphosphotidylceramide can occur; (2) introducing ceramide and phosphotylinositol, said ceramide or phosphatidylinositol carrying label for identification; (3) subjecting said active inositolphosophotidylceramide synthase, ceramide and phosphotylinositol to ordinary conditions necessary for ceramide conversion to phosphoinositol ceramide; and (4) identifying those test compounds which inhibit ceramide conversion to phosphoinositol ceramide

RFLP markers and genetic linkage of oil content and hypodermis color in sunflower seed (Helianthus annuus L.)

The service sector is dependent upon customers’ willingness to contribute their knowledge, skills, and abilities to co-produce the service experiences they want and expect. Service organizations therefore seek to employ strategies that will enhance their customers’ ability to do whatever they must to be successful in co-producing those experiences. Applying the concept of self-efficacy, we offer a theory-based approach to developing these strategies that firms may utilize. These strategies involve focusing both employee training and environmental cues on how to enhance the self-efficacy of the customer in performing whatever tasks are necessary toward a successful service experience

Technique for Specifying the Fatty Acid at the SN2 Position of Acylglycerol Lipids

A method for specifying a fatty acid at the sn2 position of acylglycerol lipids including (a) transfecting a vector including the SLC1 gene or a variant thereof into embryonic biological material, and (b) allowing the SLC1 gene to specify the type of fatty acid at the sn2 position of acylglycerol lipids. Also provided for is an isolated SLC1 gene and a probe for its detection

LAC+ Saccharomyces Cerevisiae

The invention relates to novel, transformed strains of Lac+ Saccharomyces cerevisiae, capable of utilizing lactose as a sole carbon source, produced by inserting into the Saccharomyces cerevisiae a plasmid containing a lactose permease and a beta-galactosidase gene derived from Kluyveromyces lactis yeast

Signal transduction protocols

Making Protein Immunoprecipitates Elaine A. Elion and Yunmei Wang Signal Transduction Inhibitors in Cellular Function Maofu Fu, Chenguang Wang, Xueping Zhang, and Richard G. Pestell Two-Dimensional Gel Electrophoresis for the Identification of Signaling Targets Yukihito Kabuyama, Kirsi K. Polvinen, Katheryn A. Resing, and Natalie G. Ahn A High-Throughput Mammalian Cell-Based Transient Transfection Assay Daniel J. Noonan, Kenneth Henry, and Michelle L. Twaroski Determining Protein Half-Lives Pengbo Zhou Assaying Protein Kinase Activity Jan Brabek and Steven K. Hanks Comparative Phosphorylation Site Mapping From Gel-Derived Proteins Using a Multidimensional ES/MS-Based Approach Francesca Zappacosta, Michael J. Huddleston, and Roland S. Annan Studies of Calmodulin-Dependent Regulation Paul C. Brandt and Thomas C. Vanaman Measurement of Protein-DNA Interactions In Vivo by Chromatin Immunoprecipitation Hogune Im, Jeffrey A. Grass, Kirby D. Johnson, Meghan E. Boyer, Jing Wu, and Emery H. Bresnick Characterization of Protein-DNA Association In Vivo by Chromatin Immunoprecipitation Laurent Kuras Nonradioactive Methods for Detecting Activation of Ras-Related Small G Proteins Douglas A. Andres Nucleocytoplasmic Glycosylation, O-GlcNAc: Identification and Site Mapping Natasha Elizabeth Zachara, Win Den Cheung, and Gerald Warren Hart Techniques in Protein Methylation Jaeho Lee, Donghang Cheng, and Mark T. Bedford Assaying Lipid Phosphate Phosphatase Activities Gil-Soo Han and George M. Carman Assaying Phosphoinositide Phosphatases Gregory S. Taylor and Jack E. Dixon Assaying Phospholipase A2 Activity Christina C. Leslie and Michael H. Gelb Measurement and Immunofluorescence of Cellular Phosphoinositides Hiroko Hama, Javad Torabinejad, Glenn D. Prestwich, and Daryll B. DeWald Measuring Dynamic Changes in cAMP Using Fluorescence Resonance Energy Transfer Sandrine Evellin, Marco Mongillo, Anna Terrin, Valentina Lissandron, and Manuela Zaccolo In Vivo Detection of Protein-Protein Interaction in Plant Cells Using BRET Chitra Subramanian, Yao Xu, Carl Hirschie Johnson, and Albrecht G. von Arnim Revealing Protein Dynamics by Photobleaching Techniques Frank van Drogen and Matthias Peter Assaying Cytochrome c Translocation During Apoptosis Nigel J. Waterhouse, Rohan Steel, Ruth Kluck, and Joseph A. Trapani Inde