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Stephan M Caucheteux, Vincent Piguet
Cutaneous vaccination can be a challenge because the development of local skin inflammation is often unavoidable. Thus, it is important to identify and validate new vaccine adjuvants that enhance immunization without the burden of inflammation. Wang et al. now report on a cyclic GMP-AMP adjuvant, the natural stimulator of interferon genes agonist, providing evidence for potent immune responses without inflammation.
November 2016: Journal of Investigative Dermatology
Hanan R Shehata, Cassandra L Ettinger, Jonathan A Eisen, Manish N Raizada
Endophytes are microbes that inhabit internal plant tissues without causing disease. Some endophytes are known to combat pathogens. The corn (maize) landrace Chapalote has been grown continuously by subsistence farmers in the Americas since 1000 BC, without the use of fungicides, and the crop remains highly valued by farmers, in part for its natural tolerance to pests. We hypothesized that the pathogen tolerance of Chapalote may, in part, be due to assistance from its endophytes. We previously identified a bacterial endophyte from Chapalote seeds, Burkholderia gladioli strain 3A12, for its ability to combat a diversity of crop pathogens, including Sclerotinia homoeocarpa, the most important fungal disease of creeping bentgrass, a relative of maize used here as a model system...
2016: Frontiers in Microbiology
Tatsuhiko Azegami, Yoshikazu Yuki, Kaori Hayashi, Mari Nakamura, Akihito Hishikawa, Hiroshi Kiyono, Hiroshi Itoh
OBJECTIVE: Previous studies have shown that injectable vaccines against renin-angiotensin system may be effective for the treatment of hypertension, but these injectable vaccines often cause local skin reactions and soreness at the injection site. The aim of this study was to develop a new non-injectable vaccine against angiotensin II type 1 receptor (AT1R) and examine its effect on blood pressure. DESIGN AND METHOD: The peptide, seven-amino-acid sequence from second extracellular loop of rat AT1R was synthesized and then conjugated with pneumococcal surface protein A (PspA) as a carrier protein for the nasal vaccination...
September 2016: Journal of Hypertension
Luisa F Castiblanco, George W Sundin
Bacterial biofilms are multicellular aggregates encased in an extracellular matrix mainly composed of exopolysaccharides (EPSs), protein, and nucleic acids, which determines the architecture of the biofilm. Erwinia amylovora Ea1189 forms a biofilm inside the xylem of its host, that results in vessel plugging and water transport impairment. Production of the EPSs amylovoran and levan are critical for the formation of a mature biofilm. Additionally, cyclic dimeric GMP (c-di-GMP) was reported to positively regulate amylovoran biosynthesis and biofilm formation in E...
October 18, 2016: Molecular Plant Pathology
Smita Gupta, Sunita Yadav, Venkatesan Suryanarayanan, Sanjeev K Singh, Jitendra K Saxena
Guanylate kinase is one of the key enzymes in nucleotide biosynthesis. The study highlights the structural and functional properties of Brugia malayi Guanylate kinase (BmGK) in the presence of chemical denaturants. An inactive, partially unfolded, dimeric intermediate was observed at 1-2M urea while GdnCl unfolding showed monomer molten globule like intermediate at 0.8-1.0M. The results also illustrate the protective role of substrates in maintaining the integrity of the enzyme. The thermo stability of protein was found to be significantly enhanced in the presence of the substrates...
October 14, 2016: International Journal of Biological Macromolecules
Christoph Priesner, Ruth Esser, Sabine Tischer, Michael Marburger, Krasimira Aleksandrova, Britta Maecker-Kolhoff, Hans-Gert Heuft, Lilia Goudeva, Rainer Blasczyk, Lubomir Arseniev, Ulrike Köhl, Britta Eiz-Vesper, Stephan Klöß
BACKGROUND AND AIMS: The infusion of enriched CMV-specific donor T-cells appears to be a suitable alternative for the treatment of drug-resistant CMV reactivation or de novo infection after both solid organ and hematopoietic stem cell transplantation. Antiviral lymphocytes can be selected from apheresis products using the CliniMACS Cytokine-Capture-System(®) either with the well-established CliniMACS(®) Plus (Plus) device or with its more versatile successor CliniMACS Prodigy(®) (Prodigy)...
2016: Frontiers in Immunology
Karin Förster-Fromme, Sarah Schneider, Georg A Sprenger, Christoph Albermann
OBJECTIVES: To investigate the translocation of nucleotide-activated sugars from the cytosol across a membrane into the endoplasmatic reticulum or the Golgi apparatus which is an important step in the synthesis of glycoproteins and glycolipids in eukaryotes. RESULTS: The heterologous expression of the recombinant and codon-adapted human GDP-L-fucose antiporter gene SLC35C1 (encoding an N-terminal OmpA-signal sequence) led to a functional transporter protein located in the cytoplasmic membrane of Escherichia coli...
October 13, 2016: Biotechnology Letters
Shuting Xu, Aurélie Ducroux, Aparna Ponnurangam, Gabrielle Vieyres, Sergej Franz, Mathias Müsken, Thomas Zillinger, Angelina Malassa, Ellen Ewald, Veit Hornung, Winfried Barchet, Susanne Häussler, Thomas Pietschmann, Christine Goffinet
Upon sensing cytoplasmic retroviral DNA in infected cells, cyclic GMP-AMP (cGAMP) synthase (cGAS) produces the cyclic dinucleotide cGAMP, which activates STING to trigger a type I interferon (IFN) response. We find that membrane fusion-inducing contact between donor cells expressing the HIV envelope (Env) and primary macrophages endogenously expressing the HIV receptor CD4 and coreceptor enable intercellular transfer of cGAMP. This cGAMP exchange results in STING-dependent antiviral IFN responses in target macrophages and protection from HIV infection...
October 12, 2016: Cell Host & Microbe
Ammar W Ashor, Shakir Chowdhury, Clio Oggioni, Othman Qadir, Kirsten Brandt, Abbas Ishaq, John C Mathers, Gabriele Saretzki, Mario Siervo
BACKGROUND: Aging and obesity are associated with raised oxidative stress and a reduction of nitric oxide (NO) bioavailability, with subsequent decline in insulin sensitivity and endothelial function. Inorganic nitrate is converted into NO via a 2-step reduction process and may be an effective nutritional intervention to modify vascular and metabolic functions. OBJECTIVES: This study tested whether inorganic nitrate supplementation improved glucose disposal and attenuated the acute effects of hyperglycemia on oxidative stress, inflammation, and vascular function in young and old obese participants...
October 12, 2016: Journal of Nutrition
Milan M Milutinović, Jovana V Bogojeski, Olivera Klisurić, Andreas Scheurer, Sofi K C Elmroth, Živadin D Bugarčić
A novel rhodium(iii) complex [Rh(III)(H2L(tBu))Cl3] (1) (H2L(tBu) = 2,6-bis(5-tert-butyl-1H-pyrazol-3-yl)pyridine) containing a pincer type, tridentate nitrogen-donor chelate system was synthesized. Single crystal X-ray structure analysis revealed that 1 crystallizes in the orthorhombic space group Pbcn with a = 20.7982(6), b = 10.8952(4), c = 10.9832(4) Å, V = 2488.80(15) Å(3), and eight molecules in the unit cell. The rhodium center in the complex [Rh(III)(H2L(tBu))Cl3] (1) is coordinated in a slightly distorted octahedral geometry by the tridentate N,N,N-donor and three chloro ligands, adopting a mer arrangement with an essentially planar ligand skeleton...
October 4, 2016: Dalton Transactions: An International Journal of Inorganic Chemistry
Daniel Jose Matos Medeiros-Lima, Antonio Claudio Mendes-Ribeiro, Tatiana Marlowe Cunha Brunini, Marcela Anjos Martins, Wanda Vianna Mury, Raul Almeira Freire, Walace David Monteiro, Paulo Tarso Veras Farinatti, Cristiane Matsuura
Growing evidence has shown that acute exercise impairs erythrocyte membrane structure and function as a consequence of increased physical and chemical stress. Erythrocyte-synthesized nitric oxide (NO) is known to modulate membrane fluidity, and its bioavailability depends on the balance between its production and scavenging by reactive oxygen species. Here, we investigated whether a maximal exercise test could affect erythrocyte NO bioavailability and oxidative stress. Twelve men (26±4 years old, V̇O2peak 44...
October 3, 2016: Clinical Hemorheology and Microcirculation
Christopher Poon, Xiaopin Duan, Christina Chan, Wenbo Han, Wenbin Lin
Due to the ability of ovarian cancer (OCa) to acquire drug resistance, it has been difficult to develop efficient and safe chemotherapy for OCa. Here, we examined the therapeutic use of a new self-assembled core-shell nanoscale coordination polymer nanoparticle (NCP-Carbo/GMP) that delivers high loadings of carboplatin (28.0±2.6 wt.%) and gemcitabine monophosphate (8.6±1.5 wt.%). A strong synergistic effect was observed between carboplatin and gemcitabine against platinum-resistant OCa cells, SKOV-3, and A2780/CDDP in vitro...
October 6, 2016: Molecular Pharmaceutics
A R Bustos-Carpinteyro, N Delgado-Figueroa, E Santiago-Luna, M T Magaña-Torres, J Y Sánchez-López
Gastric cancer (GC), the third leading cause of cancer-related deaths in Mexico and worldwide, can be classified into diffuse (DGC) or intestinal (IGC) types based on its histological characteristics. DGC is characterized by reduced expression of the cell adhesion protein E-cadherin, which is encoded by CDH1. The -472delA (rs5030625) and -160C>A (rs16260) polymorphisms in CDH1 induce a decrease in gene transcription; in fact, these mutated alleles have been associated with GC in some populations, with conflicting results...
September 16, 2016: Genetics and Molecular Research: GMR
Yujie Xiao, Hailing Nie, Huizhong Liu, Wenli Chen, Qiaoyun Huang
Cyclic di-GMP (c-di-GMP), a ubiquitous bacterial second messenger that regulates diverse cellular processes, is synthesized by diguanylate cyclase (DGC) and degraded by phosphodiesterase (PDE). GcbA is a well conserved DGC among Pseudomonas species, and has been reported to influence biofilm formation and flagellar motility in Pseudomonas fluorescens and Pseudomonas aeruginosa. Here we confirm the function of GcbA in Pseudomonas putida and reveal that expression of GcbA is regulated by FleQ in response to c-di-GMP...
October 4, 2016: Environmental Microbiology Reports
Mona W Orr, Michael Y Galperin, Vincent T Lee
Bacteria utilize a diverse set of nucleotide second messengers to regulate cellular responses by binding macromolecular receptors (RNAs and proteins). Recent studies on cyclic di-GMP (c-di-GMP) have shown that this signaling molecule binds multiple receptors to regulate different steps in the same biological process. We propose this property of the same molecule regulating multiple steps in the same process is biologically meaningful and have termed this phenomenon 'sustained sensing'. Here, we discuss the recent findings that support the concept of sustained sensing of c-di-GMP levels and provide additional examples that support the utilization of sustained sensing by other second messengers...
October 1, 2016: Current Opinion in Microbiology
Kwang-Jin Cho, Darren E Casteel, Priyanka Prakash, Lingxiao Tan, Dharini van der Hoeven, Angela A Salim, Choel Kim, Robert J Capon, Ernest Lacey, Shane R Cunha, Alemayehu A Gorfe, John F Hancock
K-Ras must localize to the plasma membrane and be arrayed in nanoclusters for biological activity. We show here that K-Ras is a substrate for cyclic GMP-dependent protein kinases (PKGs). In intact cells activated PKG2 selectively co-localizes with K-Ras on the plasma membrane and phosphorylates K-Ras at Ser181 in the C-terminal polybasic domain. K-Ras phosphorylation by PKG2 is triggered by activation of AMP kinase and requires eNOS and soluble guanylyl-cyclase. Phosphorylated K-Ras re-organizes into distinct nanoclusters that retune signal output...
October 3, 2016: Molecular and Cellular Biology
Rameshwar K Sharma, Teresa Duda, Clint L Makino
This monograph presents a historical perspective of cornerstone developments on the biochemistry and physiology of mammalian membrane guanylate cyclases (MGCs), highlighting contributions made by the authors and their collaborators. Upon resolution of early contentious studies, cyclic GMP emerged alongside cyclic AMP, as an important intracellular second messenger for hormonal signaling. However, the two signaling pathways differ in significant ways. In the cyclic AMP pathway, hormone binding to a G protein coupled receptor leads to stimulation or inhibition of an adenylate cyclase, whereas the cyclic GMP pathway dispenses with intermediaries; hormone binds to an MGC to affect its activity...
2016: Frontiers in Molecular Neuroscience
Bart W Faber, Stephan Hellwig, Sophie Houard, Nicolas Havelange, Jürgen Drossard, Hubert Mertens, Alexander Croon, Robin Kastilan, Richard Byrne, Nicole van der Werff, Marjolein van der Eijk, Alan W Thomas, Clemens H M Kocken, Edmond J Remarque
Plasmodium falciparum apical membrane antigen 1 (PfAMA1) is a leading asexual blood stage vaccine candidate for malaria. In preparation for clinical trials, three Diversity Covering (DiCo) PfAMA1 ectodomain proteins, designed to overcome the intrinsic polymorphism that is present in PfAMA1, were produced under Good Manufacturing Practice (GMP) in Pichia pastoris. Using identical methodology, the 3 strains were cultivated in 70-L scale fed-batch fermentations and PfAMA1-DiCos were purified by two chromatography steps, an ultrafiltration/diafiltration procedure and size exclusion chromatography, resulting in highly pure (>95%) PfAMA1-DiCo1, PfAMA1 DiCo2 and PfAMA1 DiCo3, with final yields of 1...
2016: PloS One
Emily A Sawin, Bridget M Stroup, Sangita G Murali, Lucas M O'Neill, James M Ntambi, Denise M Ney
BACKGROUND: Glycomacropeptide (GMP) is a 64-amino acid glycophosphopeptide released from κ-casein during cheesemaking that promotes satiety, reduces body fat, increases bone mass and infers prebiotic and anti-inflammatory effects. The impact of adiposity and gender on bone health is unclear. OBJECTIVE: To determine how feeding female mice diets providing 60% Fat Kcal (high-fat) or 13% Fat Kcal (control) with either GMP or casein as the protein source impacts: body composition, ex vivo fatty acid oxidation, bone (femoral) biomechanical performance, and the relationship between body composition and bone...
2016: PloS One
Geneviève Pépin, Charlotte Nejad, Belinda J Thomas, Jonathan Ferrand, Kate McArthur, Philip G Bardin, Bryan R G Williams, Michael P Gantier
Acridine dyes, including proflavine and acriflavine, were commonly used as antiseptics before the advent of penicillins in the mid-1940s. While their mode of action on pathogens was originally attributed to their DNA intercalating activity, work in the early 1970s suggested involvement of the host immune responses, characterized by induction of interferon (IFN)-like activities through an unknown mechanism. We demonstrate here that sub-toxic concentrations of a mixture of acriflavine and proflavine instigate a cyclic-GMP-AMP (cGAMP) synthase (cGAS)-dependent type-I IFN antiviral response...
September 29, 2016: Nucleic Acids Research
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