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https://www.readbyqxmd.com/read/27890799/enhancing-gilthead-seabream-immune-status-and-protection-against-bacterial-challenge-by-means-of-antigens-derived-from-vibrio-parahaemolyticus
#1
Martha Reyes-Becerril, Crystal Guluarte, Diana Ceballos-Francisco, Carlos Angulo, M Ángeles Esteban
In an attempt to control the proliferation of the pathogenic bacterium Vibrio parahaemolyticus in gilthead seabream (Sparus aurata), the immunostimulant effect of lysate and ToxA from this bacterium was evaluated. Fish were intraperitoneally injected twice (first injection, day 1 of the experiment; second injection, day 7) and sampled after one week (on days 8 and 15). Afterwards, all fish specimens were experimentally infected with V. parahaemolyticus and mortality was recovered for 1 week. Fish injected with lysate, ToxA and phosphate buffer saline (control) showed 100%, 50% and 0% survival, respectively, when challenged with the pathogen...
November 24, 2016: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/27880838/detection-of-endotoxin-contamination-of-graphene-based-materials-using-the-tnf-%C3%AE-expression-test-and-guidelines-for-endotoxin-free-graphene-oxide-production
#2
Sourav P Mukherjee, Neus Lozano, Melanie Kucki, Antonio E Del Rio-Castillo, Leon Newman, Ester Vázquez, Kostas Kostarelos, Peter Wick, Bengt Fadeel
Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin detection in graphene based material (GBM) samples, including graphene oxide (GO) and few-layered graphene (FLG). Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET) using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay...
2016: PloS One
https://www.readbyqxmd.com/read/27878261/bacterial-ghosts-as-adjuvants-in-syngeneic-tumour-cell-lysate-based-anticancer-vaccination-in-a-murine-lung-carcinoma-model
#3
Jan Aleksander Kraśko, Karolina Žilionytė, Adas Darinskas, Marius Strioga, Svetlana Rjabceva, Iosif Zalutsky, Marina Derevyanko, Vladimir Kulchitsky, Werner Lubitz, Pavol Kudela, Edita Miseikyte-Kaubriene, Olha Karaman, Hennadii Didenko, Hryhorii Potebnya, Vasyl Chekhun, Vita Pašukonienė
Instead of relying on external anticancer factors for treatment, immunotherapy utilizes the host's own immune system and directs it against given tumour antigens. This study demonstrated that it is possible to overcome the documented immunosuppressive properties of tumour cell lysate by supplementing it with appropriate adjuvant. Lewis lung carcinoma (LLC)‑challenged C57BL/6 mice were treated with LLC cryo‑lysate mixed with either bacterial ghosts (BGs) generated from E. coli Nissle 1917 or B. subtilis 70 kDa protein as adjuvants...
November 16, 2016: Oncology Reports
https://www.readbyqxmd.com/read/27864613/oncolysate-loaded-escherichia-coli-bacterial-ghosts-enhance-the-stimulatory-capacity-of-human-dendritic-cells
#4
Jaroslav Michalek, Renata Hezova, Pavlina Turanek-Knötigova, Jana Gabkova, Marius Strioga, Werner Lubitz, Pavol Kudela
The natural adjuvant properties of bacterial ghosts (BGs) lie within the presence of intact pathogen-associated molecular patterns on their surface. BGs can improve the direct delivery, natural processing and presentation of target antigens within dendritic cells (DCs). Moreover, sensitization of human DCs by cancer cell lysate (oncolysate)-loaded BGs in the presence of IFN-α and GM-CSF enhanced DC maturation as indicated by an increased expression of maturation markers and co-stimulatory molecules, higher production of IL-12p70 and stimulation of significantly increased proliferation of both autologous CD4(+) and CD8(+) T cells compared to DCs matured in the presence of purified lipopolysaccharide...
November 18, 2016: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/27864297/expression-of-biologically-active-murine-interleukin-18-il-18-in-lactococcus-lactis
#5
Sadegh Feizollahzadeh, Hossein Khanahmad, Ilnaz Rahimmanesh, Mazdak Ganjalikhani-Hakemi, Alireza Andalib, Mohammad Hossein Sanei, Abbas Rezaei
Food grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L...
November 17, 2016: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/27854308/characterization-of-chemically-induced-bacterial-ghosts-bgs-using-sodium-hydroxide-induced-vibrio-parahaemolyticus-ghosts-vpgs
#6
Hyun Jung Park, Sung Oh, Nagarajan Vinod, Seongmi Ji, Han Byul Noh, Jung Mo Koo, Su Hyeong Lee, Sei Chang Kim, Ki-Sung Lee, Chang Won Choi
Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene E of bacteriophage PhiX174. In this study, Vibrio parahaemolyticus ghosts (VPGs) were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid, and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6...
November 15, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27844185/a-j-modulated-protonless-nmr-experiment-characterizes-the-conformational-ensemble-of-the-intrinsically-disordered-protein-wip
#7
Eva Rozentur-Shkop, Gil Goobes, Jordan H Chill
Intrinsically disordered proteins (IDPs) are multi-conformational polypeptides that lack a single stable three-dimensional structure. It has become increasingly clear that the versatile IDPs play key roles in a multitude of biological processes, and, given their flexible nature, NMR is a leading method to investigate IDP behavior on the molecular level. Here we present an IDP-tailored J-modulated experiment designed to monitor changes in the conformational ensemble characteristic of IDPs by accurately measuring backbone one- and two-bond J((15)N,(13)Cα) couplings...
November 14, 2016: Journal of Biomolecular NMR
https://www.readbyqxmd.com/read/27825951/a-midgut-lysate-of-the-riptortus-pedestris-has-antibacterial-activity-against-lps-o-antigen-deficient-burkholderia-mutants
#8
Ho Am Jang, Eun Sil Seo, Min Young Seong, Bok Luel Lee
Riptortus pedestris, a common pest in soybean fields, harbors a symbiont Burkholderia in a specialized posterior midgut region of insects. Every generation of second nymphs acquires new Burkholderia cells from the environment. We compared in vitro cultured Burkholderia with newly in vivo colonized Burkholderia in the host midgut using biochemical approaches. The bacterial cell envelope of in vitro cultured and in vivo Burkholderia differed in structure, as in vivo bacteria lacked lipopolysaccharide (LPS) O-antigen...
November 5, 2016: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/27822540/identification-of-novel-host-interactors-of-effectors-secreted-by-salmonella-and-citrobacter
#9
Ryan L Sontag, Ernesto S Nakayasu, Roslyn N Brown, George S Niemann, Michael A Sydor, Octavio Sanchez, Charles Ansong, Shao-Yeh Lu, Hyungwon Choi, Dylan Valleau, Karl K Weitz, Alexei Savchenko, Eric D Cambronne, Joshua N Adkins
Many pathogenic bacteria of the family Enterobacteriaceae use type III secretion systems to inject virulence proteins, termed "effectors," into the host cell cytosol. Although host-cellular activities of several effectors have been demonstrated, the function and host-targeted pathways of most of the effectors identified to date are largely undetermined. To gain insight into host proteins targeted by bacterial effectors, we performed coaffinity purification of host proteins from cell lysates using recombinant effectors from the Enterobacteriaceae intracellular pathogens Salmonella enterica serovar Typhimurium and Citrobacter rodentium...
July 2016: MSystems
https://www.readbyqxmd.com/read/27775835/clinical-efficacy-of-edta-ultrasonic-activation-in-the-reduction-of-endotoxins-and-cultivable-bacteria
#10
D R Herrera, F C Martinho, A J de-Jesus-Soares, A A Zaia, C C Ferraz, J F Almeida, B P Gomes
AIM: This clinical study was conducted to investigate the influence of 17% ethylenediaminetetraacetic acid (EDTA) ultrasonic activation after chemomechanical preparation (CMP) on eliminating/reducing oral bacterial lipopolysaccharides (known as endotoxins) and cultivable bacteria in teeth with pulp necrosis and apical periodontitis. METHODOLOGY: Samples were taken from 24 root canals at several clinical periods: S1- before CMP; S2- after CMP; S3- after EDTA: G1- with ultrasonic activation (n=12), and G2- without ultrasonic activation (n=12)...
October 24, 2016: International Endodontic Journal
https://www.readbyqxmd.com/read/27773849/intranasal-administration-of-a-polyvalent-bacterial-lysate-induces-self-restricted-inflammation-in-the-lungs-and-a-th1-th17-memory-signature
#11
Analía Rial, Florencia Ferrara, Norma Suárez, Paola Scavone, Juan Martín Marqués, José Alejandro Chabalgoity
Respiratory tract infections are among the most frequent infections in humans causing millions of deaths especially in children and the elderly. Antibiotics and vaccines are the main available tools of control, but resistant strains are continuously arising and available vaccines only account for few of many pathogens involved. Non-specific immunotherapies are an emerging alternative to induce protective immunity at the airways. Mucosally administered polyvalent bacterial lysates (PBLs) have been widely used for decades for prevention of respiratory diseases, but the bases of their proposed therapeutic effectiveness are still controversial...
October 20, 2016: Microbes and Infection
https://www.readbyqxmd.com/read/27769383/rapid-detection-of-bacterial-endotoxins-in-ophthalmic-viscosurgical-device-materials-by-direct-analysis-in-real-time-mass-spectrometry
#12
Hongli Li, Victoria M Hitchins, Samanthi Wickramasekara
Bacterial endotoxins are lipopolysaccharides bound to the bacterial cell wall and released when bacteria rupture or disintegrate. Possible contamination of endotoxin in ophthalmic devices can cause a painful eye inflammation or result in toxic anterior segment syndrome after cataract surgery. Measurement of bacterial endotoxin in medical device materials is difficult since endotoxin binds with polymer matrix and some of the materials are very viscous and non-water soluble, where traditional enzyme-based Limulus amebocyte lysate (LAL) assay cannot be applied...
November 2, 2016: Analytica Chimica Acta
https://www.readbyqxmd.com/read/27768741/a-simple-and-rapid-method-for-preparing-a-cell-free-bacterial-lysate-for-protein-synthesis
#13
Nitzan Krinsky, Maya Kaduri, Janna Shainsky-Roitman, Mor Goldfeder, Eran Ivanir, Itai Benhar, Yuval Shoham, Avi Schroeder
Cell-free protein synthesis (CFPS) systems are important laboratory tools that are used for various synthetic biology applications. Here, we present a simple and inexpensive laboratory-scale method for preparing a CFPS system from E. coli. The procedure uses basic lab equipment, a minimal set of reagents, and requires less than one hour to process the bacterial cell mass into a functional S30-T7 extract. BL21(DE3) and MRE600 E. coli strains were used to prepare the S30-T7 extract. The CFPS system was used to produce a set of fluorescent and therapeutic proteins of different molecular weights (up to 66 kDa)...
2016: PloS One
https://www.readbyqxmd.com/read/27756565/purification-and-characterization-of-pseudomonas-aeruginosa-lasr-expressed-in-acyl-homoserine-lactone-free-escherichia-coli-cultures
#14
Andrés Corral Lugo, Abdelali Daddaoua, Alvaro Ortega, Bertrand Morel, Ana Isabel Díez Peña, Manuel Espinosa-Urgel, Tino Krell
Quorum sensing systems are essential for bacterial communication. We report here the purification and characterization of the Pseudomonas aeruginosa LasR quorum sensing regulator purified from lysates of E. coli cultures grown in the absence of added acyl-homoserine lactones (AHL). We show by isothermal titration calorimetry that LasR recognizes different AHLs with an affinity of approximately 1 μM. The affinity of LasR for its cognate 3-Oxo-C12-AHL was similar to that of other AHLs, indicating that this regulator has not evolved to preferentially recognize its cognate AHL...
October 15, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/27756533/immunogencity-of-antigens-from-mycobacterium-tuberculosis-self-assembled-as-particulate-vaccines
#15
Patricia Rubio Reyes, Natalie A Parlane, D Neil Wedlock, Bernd H A Rehm
Traditional approaches to vaccine development have failed to identify better vaccines to replace or supplement BCG for the control of tuberculosis (TB). Subunit vaccines offer a safer and more reproducible alternative for the prevention of diseases. In this study, the immunogenicity of bacterially derived polyester beads displaying three different Rv antigens of Mycobacterium tuberculosis was evaluated. Polyester beads displaying the antigens Rv1626, Rv2032, Rv1789, respectively, were produced in an endotoxin-free Escherichia coli strain...
October 13, 2016: International Journal of Medical Microbiology: IJMM
https://www.readbyqxmd.com/read/27756125/engineering-specificity-from-broad-to-narrow-design-of-a-blip-variant-that-exclusively-binds-and-detects-kpc-%C3%AE-lactamase
#16
Dar-Chone Chow, Kacie Rice, Wanzhi Huang, Robert L Atmar, Timothy G Palzkill
The β-lactamase inhibitory protein (BLIP) binds and inhibits a wide range of class A β-lactamases including the TEM-1 β-lactamase (Ki= 0.5 nM), which is widely present in Gram-negative bacteria, and the KPC-2 β-lactamase (Ki= 1.2 nM), which hydrolyzes virtually all clinically useful β-lactam antibiotics. The extent to which the specificity of a protein that binds a broad range of targets can be modified to display narrow specificity was explored in this study by engineering BLIP to bind selectively to KPC-2 β-lactamase...
October 18, 2016: ACS Infectious Diseases
https://www.readbyqxmd.com/read/27730561/measuring-protein-protein-interactions-using-biacore
#17
Paul Leonard, Stephen Hearty, Hui Ma, Richard O'Kennedy
The use of optical biosensors for studying macromolecular interactions is gaining increasing popularity. In one study, 1514 papers that involved the application of biosensor data were identified for the year 2009 alone (Rich and Myszka, J Mol Recognit 24:892-914, 2011), the sheer volume and variety of which present a daunting task for the burgeoning biosensor user to accumulate and decipher. This chapter is designed to provide the reader with the tools necessary to prepare, design, and efficiently execute a kinetic experiment on Biacore...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27725776/leukocyte-immunoglobulin-like-receptor-b4-regulates-key-signalling-molecules-involved-in-fc%C3%AE-ri-mediated-clathrin-dependent-endocytosis-and-phagocytosis
#18
Mijeong Park, Mark J Raftery, Paul S Thomas, Carolyn L Geczy, Katherine Bryant, Nicodemus Tedla
FcγRI cross-linking on monocytes may trigger clathrin-mediated endocytosis, likely through interaction of multiple intracellular molecules that are controlled by phosphorylation and dephosphorylation events. However, the identity of phospho-proteins and their regulation are unknown. We proposed the leukocyte immunoglobulin-like receptor B4 (LILRB4) that inhibits FcγRI-mediated cytokine production via Tyr dephosphorylation of multiple kinases, may also regulate endocytosis/phagocytosis through similar mechanisms...
October 11, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27725208/whole-cell-biosensor-of-cellobiose-and-application-to-wood-decay-detection
#19
Maxime Toussaint, Cyril Bontemps, Arnaud Besserer, Laurence Hotel, Philippe Gérardin, Pierre Leblond
Fungal biodegradation of wood is one of the main threats regarding its use as a material. So far, the detection of this decaying process is empirically assessed by loss of mass, when the fungal attack is advanced and woody structure already damaged. Being able to detect fungal attack on wood in earlier steps is thus of special interest for the wood economy. In this aim, we designed here a new diagnostic tool for wood degradation detection based on the bacterial whole-cell biosensor technology. It was designed in diverting the soil bacteria Streptomyces CebR sensor system devoted to cellobiose detection, a cellulolytic degradation by-product emitted by lignolytic fungi since the onset of wood decaying process...
October 8, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27707587/bacterial-extract-om-85-with-human-equivalent-doses-does-not-inhibit-the-development-of-asthma-in-a-murine-model
#20
A Rodrigues, L P Gualdi, R G de Souza, M H M Vargas, N K Nuñez, A A da Cunha, M H Jones, L A Pinto, R T Stein, P M Pitrez
BACKGROUND: OM-85 is an immunostimulant bacterial lysate, which has been proven effective in reducing the number of lower airways infections. We investigated the efficacy of the bacterial lysate OM-85 in the primary prevention of a murine model of asthma. METHODS: In the first phase of our study the animals received doses of 0.5μg, 5μg and 50μg of OM-85 through gavage for five days (days -10 to -6 of the protocol), 10 days prior to starting the sensitisation with ovalbumin (OVA), in order to evaluate the results of dose-response protocols...
October 1, 2016: Allergologia et Immunopathologia
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