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https://www.readbyqxmd.com/read/27914091/toward-clinical-use-of-the-igg-specific-enzymes-ides-and-endos-against-antibody-mediated-diseases
#1
Mattias Collin, Lars Björck
The endoglycosidase EndoS and the protease IdeS from the human pathogen Streptococcus pyogenes are immunomodulating enzymes hydrolyzing human IgG. IdeS cleaves IgG in the lower hinge region, while EndoS hydrolyzes the conserved N-linked glycan in the Fc region. Both enzymes are remarkably specific for human IgG that after hydrolysis loses most of its effector functions, such as binding to leukocytes and complement activation, all contributing to bacterial evasion of adaptive immunity. However, taken out of their infectious context, we and others have shown that IdeS and EndoS can alleviate autoimmune disease in a number of animal models of antibody-mediated disorders...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914089/generating-and-purifying-fab-fragments-from-human-and-mouse-igg-using-the-bacterial-enzymes-ides-speb-and-kgp
#2
Jonathan Sjögren, Linda Andersson, Malin Mejàre, Fredrik Olsson
Fab fragments are valuable research tools in various areas of science including applications in imaging, binding studies, removal of Fc-mediated effector functions, mass spectrometry, infection biology, and many others. The enzymatic tools for the generation of Fab fragments have been discovered through basic research within the field of molecular bacterial pathogenesis. Today, these enzymes are widely applied as research tools and in this chapter, we describe methodologies based on bacterial enzymes to generate Fab fragments from both human and mouse IgG...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914082/detection-of-intracellular-proteins-by-high-resolution-immunofluorescence-microscopy-in-streptococcus-pyogenes
#3
Assaf Raz
Immunofluorescence microscopy is an invaluable tool for the study of biological processes at the cellular level. While the localization of surface-exposed antigens can easily be determined using fluorescent antibodies, localization of intracellular antigens requires permeabilization of the bacterial cell wall and membrane. Here, we describe an immunofluorescence protocol tailored specifically for Streptococcus pyogenes, applying the phage lysin PlyC for cell wall permeabilization. This protocol allows a high level of morphological preservation, suitable for high-resolution microscopy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27913674/immunoglobulins-and-their-receptors-and-subversion-of-their-protective-roles-by-bacterial-pathogens
#4
REVIEW
Jenny M Woof
Immunoglobulins (Igs) play critical roles in immune defence against infectious disease. They elicit potent elimination processes such as triggering complement activation and engaging specific Fc receptors present on immune cells, resulting in phagocytosis and other killing mechanisms. Many important pathogens have evolved mechanisms to subvert or evade Ig-mediated defence. One such mechanism used by several pathogenic bacteria features proteins that bind the Ig Fc region and compromise engagement of host effector molecules...
December 15, 2016: Biochemical Society Transactions
https://www.readbyqxmd.com/read/27900888/using-the-crispr-cas9-system-to-eliminate-native-plasmids-of-zymomonas-mobilis-zm4
#5
Qing-Hua Cao, Huan-Huan Shao, Hui Qiu, Tao Li, Yi-Zheng Zhang, Xue-Mei Tan
The CRISPR/Cas system can be used to simply and efficiently edit the genomes of various species, including animals, plants, and microbes. Zymomonas mobilis ZM4 is a highly efficient, ethanol-producing bacterium that contains five native plasmids. Here, we constructed the pSUZM2a-Cas9 plasmid and a single-guide RNA expression plasmid. The pSUZM2a-Cas9 plasmid was used to express the Cas9 gene cloned from Streptococcus pyogenes CICC 10464. The single-guide RNA expression plasmid pUC-T7sgRNA, with a T7 promoter, can be used for the in vitro synthesis of single-guide RNAs...
November 30, 2016: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/27895128/characterization-of-the-pathogenicity-of-streptococcus-intermedius-tyg1620-isolated-from-a-human-brain-abscess-based-on-the-complete-genome-sequence-with-transcriptome-analysis-and-transposon-mutagenesis-in-a-murine-subcutaneous-abscess-model
#6
Noriko Hasegawa, Tsuyoshi Sekizuka, Yutaka Sugi, Nobuhiro Kawakami, Yumiko Ogasawara, Kengo Kato, Akifumi Yamashita, Fumihiko Takeuchi, Makoto Kuroda
Streptococcus intermedius is known to cause periodontitis and pyogenic infections in the brain and liver. Here we report the complete genome sequence of strain TYG1620 (genome size: 2,006,877 bp; GC content: 37.6%; 2,020 predicted ORFs) isolated from a brain abscess in an infant. Comparative genome analysis of S. intermedius genome sequences suggested that TYG1620 carries a notable type VII secretion system (T7SS), two long-repeat regions and 19 ORFs for cell-wall-anchored proteins (CWAPs). To elucidate genes responsible for the pathogenicity of TYG1620, transcriptome analysis was performed in a murine subcutaneous abscess model...
November 28, 2016: Infection and Immunity
https://www.readbyqxmd.com/read/27874935/effect-of-administration-of-streptococcus-salivarius-k12-on-the-occurrence-of-streptococcal-pharyngo-tonsillitis-scarlet-fever-and-acute-otitis-media-in-3-years-old-children
#7
F Di Pierro, M Colombo, M G Giuliani, M L Danza, I Basile, T Bollani, A M Conti, A Zanvit, A S Rottoli
OBJECTIVE: Streptococcus salivarius K12 (BLIS K12) is a probiotic strain strongly antagonistic to the growth of Streptococcus pyogenes, the most important bacterial cause of pharyngeal infections in humans. Shown to colonize the oral cavity and to be safe for human use, BLIS K12 has previously been reported to reduce pharyngo-tonsillitis episodes in children or adults known to have experienced recurrent streptococcal infection. The present study was focussed upon evaluating the role of BLIS K12 in the control of streptococcal disease and acute otitis media in children attending the first year of kindergarten...
November 2016: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/27874072/cas9-catalyzed-dna-cleavage-generates-staggered-ends-evidence-from-molecular-dynamics-simulations
#8
Zhicheng Zuo, Jin Liu
The CRISPR-associated endonuclease Cas9 from Streptococcus pyogenes (spCas9) along with a single guide RNA (sgRNA) has emerged as a versatile toolbox for genome editing. Despite recent advances in the mechanism studies on spCas9-sgRNA-mediated double-stranded DNA (dsDNA) recognition and cleavage, it is still unclear how the catalytic Mg(2+) ions induce the conformation changes toward the catalytic active state. It also remains controversial whether Cas9 generates blunt-ended or staggered-ended breaks with overhangs in the DNA...
November 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27868255/regulatory-gene-mutation-a-driving-force-behind-group-a-streptococcus-strain-and-serotype-specific-variation
#9
Poulomee Sarkar, Paul Sumby
Data from multiple bacterial pathogens are consistent with regulator-encoding genes having higher mutation frequencies than the genome average. Such mutations drive both strain- and type- (e.g. serotype, haplotype) specific phenotypic heterogeneity, and may challenge public health due to the potential of variants to circumvent established treatment and/or preventative regimes. Here, using the human bacterial pathogen the group A Streptococcus (GAS; S. pyogenes) as a model organism, we review the types and regulatory-, phenotypic-, and disease-specific consequences of naturally occurring regulatory gene mutations...
November 21, 2016: Molecular Microbiology
https://www.readbyqxmd.com/read/27864476/anti-hinge-antibodies-recognize-igg-subclass-and-protease-restricted-neoepitopes
#10
Willem J J Falkenburg, Dirkjan van Schaardenburg, Pleuni Ooijevaar-de Heer, Michel W P Tsang-A-Sjoe, Irene E M Bultink, Alexandre E Voskuyl, Arthur E H Bentlage, Gestur Vidarsson, Gertjan Wolbink, Theo Rispens
Anti-hinge Abs (AHAs) target neoepitopes exposed after proteolytic cleavage of IgG. In this study, we explored the diversity of protease- and IgG subclass-restricted AHAs and their potential as immunological markers in healthy donors (HDs) and patients with rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE). AHA reactivity against IgG-degrading enzyme of Streptococcus pyogenes (IdeS)- or pepsin-generated F(ab')2 fragments of all four human IgG subclasses was determined. AHA reactivity against one or more out of eight F(ab')2 targets was found in 68% (68 of 100) of HDs, 69% (68 of 99) of SLE patients, and 81% (79 of 97) of RA patients...
November 18, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/27862457/a-pts-eii-mutant-library-in-group-a-streptococcus-identifies-a-promiscuous-man-family-pts-transporter-influencing-sls-mediated-hemolysis
#11
Ganesh S Sundar, Emrul Islam, Kanika Gera, Yoann Le Breton, Kevin S McIver
The Group A Streptococcus (GAS, Streptococcus pyogenes) is a Gram-positive human pathogen that must adapt to unique host environments in order to survive. Links between sugar metabolism and virulence have been demonstrated in GAS, where mutants in the phosphoenolpyruvate-dependent phosphotransferase system (PTS) exhibited Streptolysin S (SLS)-mediated hemolysis during exponential growth. This early onset hemolysis correlated with an increased lesion size and severity in a murine soft tissue infection model when compared with parental M1T1 MGAS5005...
November 8, 2016: Molecular Microbiology
https://www.readbyqxmd.com/read/27862322/staphylococcus-aureus-isolated-from-tonsillectomized-adult-patients-with-recurrent-tonsillitis
#12
Marta Katkowska, Katarzyna Garbacz, Józef Stromkowski
The aim of this study was to analyze the prevalence and antibiotic resistance of Staphylococcus aureus strains from 118 tonsillectomized adults due to recurrent tonsillitis (RT). The study included strains isolated from the tonsillar surface prior to tonsillectomy, recovered from the tonsillar core at the time of surgery, and from the posterior throat 2-4 weeks after the procedure. Susceptibility of isolates to 19 antibiotics was tested in line with the Clinical and Laboratory Standards Institute recommendations...
November 16, 2016: APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica
https://www.readbyqxmd.com/read/27845387/adenoviral-vectors-encoding-crispr-cas9-multiplexes-rescue-dystrophin-synthesis-in-unselected-populations-of-dmd-muscle-cells
#13
Ignazio Maggio, Jin Liu, Josephine M Janssen, Xiaoyu Chen, Manuel A F V Gonçalves
Mutations disrupting the reading frame of the ~2.4 Mb dystrophin-encoding DMD gene cause a fatal X-linked muscle-wasting disorder called Duchenne muscular dystrophy (DMD). Genome editing based on paired RNA-guided nucleases (RGNs) from CRISPR/Cas9 systems has been proposed for permanently repairing faulty DMD loci. However, such multiplexing strategies require the development and testing of delivery systems capable of introducing the various gene editing tools into target cells. Here, we investigated the suitability of adenoviral vectors (AdVs) for multiplexed DMD editing by packaging in single vector particles expression units encoding the Streptococcus pyogenes Cas9 nuclease and sequence-specific gRNA pairs...
November 15, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27843558/prevalence-of-bovine-milk-pathogens-in-azorean-pastures-mobile-versus-fixed-milking-machines
#14
C Azevedo, D Pacheco, L Soares, M Moitoso, J Maldonado, R Guix, J Simões
The aims of the present study were (1) to evaluate the influence of using mobile (n=47) or fixed (n=45) milking machines in Azorean herds on the apparent prevalence of several milk pathogens in bulk tank milk (BTM) and (2) to determine whether separated subclinical mastitic cows can serve, in real time, as predictors of milk pathogen prevalence for the remaining animals at the herd level. The use of a mobile or fixed milking machine influenced (P≤0.05) the prevalence of Staphylococcus aureus (72.3 per cent; n=34 v 51...
2016: Veterinary Record Open
https://www.readbyqxmd.com/read/27842538/the-modified-gingyo-san-a-chinese-herbal-medicine-has-direct-antibacterial-effects-against-respiratory-pathogens
#15
Tetsuya Yamada, Takeaki Wajima, Hidemasa Nakaminami, Kaho Kobayashi, Hideaki Ikoshi, Norihisa Noguchi
BACKGROUND: Modified Gingyo-san (MGS) is empirically used to treat various respiratory infections. MGS has been reported to have antiinflammatory and antiviral activities; however, it is not known if it has an antibacterial activity. Therefore, in this study, we aimed to investigate the antimicrobial activity of MGS against respiratory pathogens. METHODS: MGS, which is sold as an over-the-counter drug in Japan, was used for the study. Antimicrobial activity was evaluated using the disk diffusion method...
November 14, 2016: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/27832543/using-an-inducible-crispr-dcas9-krab-effector-system-to-dissect-transcriptional-regulation-in-human-embryonic-stem-cells
#16
Krishna Mohan Parsi, Erica Hennessy, Nicola Kearns, René Maehr
CRISPR-Cas9 effector systems have wide applications for the stem cell and regenerative medicine field. The ability to dissect the functional gene regulatory networks in pluripotency and potentially in differentiation intermediates of all three germ layers makes this a valuable tool for the stem cell community. Catalytically inactive Cas9 fused to transcriptional/chromatin effector domains allows for silencing or activation of a genomic region of interest. Here, we describe the application of an inducible, RNA-guided, nuclease-deficient (d) Cas9-KRAB system (adapted from Streptococcus pyogenes) to silence target gene expression in human embryonic stem cells, via KRAB repression at the promoter region...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27830834/suilysin-induced-platelet-neutrophil-complexes-formation-is-triggered-by-pore-formation-dependent-calcium-influx
#17
Shengwei Zhang, Yuling Zheng, Shaolong Chen, Shujing Huang, Keke Liu, Qingyu Lv, Yongqiang Jiang, Yuan Yuan
Platelet activation and platelet-neutrophil interactions have been found to be involved in inflammation, organ failure and soft-tissue necrosis in bacterial infections. Streptococcus suis, an emerging human pathogen, can cause streptococcal toxic-shock syndrome (STSS) similarly to Streptococcus pyogenes. Currently, S. suis-platelet interactions are poorly understood. Here, we found that suilysin (SLY), the S. suis cholesterol-dependent cytolysin (CDC), was the sole stimulus of S. suis that induced platelet-neutrophil complexes (PNC) formation...
November 10, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27826373/acute-cardiac-tamponade-in-a-58-year-old-male-with-poststreptococcal-glomerulonephritis
#18
Wendy Bottinor, Daniel Fronk, Salima Sadruddin, Harriet Foster, Nilang Patel, Andreas Prinz, Ion S Jovin
Pericarditis in conjunction with nephritis is an uncommon clinical scenario with a broad differential diagnosis. We present the case of a 58-year-old male who developed nephritis and pericardial effusion with tamponade physiology. In the following, we discuss the differential diagnosis for concomitant nephritis and pericarditis and discuss the work-up performed on our patient. We also review the epidemiology of postinfectious glomerulonephritis in adults and describe previous cases of Streptococcus pyogenes pericarditis in the literature...
September 2016: Methodist DeBakey Cardiovascular Journal
https://www.readbyqxmd.com/read/27826289/pimms-pragmatic-insertional-mutation-mapping-system-laboratory-methodology-a-readily-accessible-tool-for-identification-of-essential-genes-in-streptococcus
#19
Adam M Blanchard, Sharon A Egan, Richard D Emes, Andrew Warry, James A Leigh
The Pragmatic Insertional Mutation Mapping (PIMMS) laboratory protocol was developed alongside various bioinformatics packages (Blanchard et al., 2015) to enable detection of essential and conditionally essential genes in Streptococcus and related bacteria. This extended the methodology commonly used to locate insertional mutations in individual mutants to the analysis of mutations in populations of bacteria. In Streptococcus uberis, a pyogenic Streptococcus associated with intramammary infection and mastitis in ruminants, the mutagen pGhost9:ISS1 was shown to integrate across the entire genome...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27821851/phenotypic-differentiation-of-streptococcus-pyogenes-populations-is-induced-by-recombination-driven-gene-specific-sweeps
#20
Yun-Juan Bao, B Jesse Shapiro, Shaun W Lee, Victoria A Ploplis, Francis J Castellino
Genomic recombination plays an important role in driving adaptive evolution and population differentiation in bacteria. However, controversy exists as to the effects of recombination on population diversity and differentiation, i.e., recombination is frequent enough to sweep through the population at selected gene loci (gene-specific sweeps), or the recombination rate is low without interfering genome-wide selective sweeps. Observations supporting either view are sparse. Pathogenic bacteria causing infectious diseases are promising candidates to provide observations of recombination...
November 8, 2016: Scientific Reports
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