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"neisseria meningitidis"

Xianyang Wang, Peng Wang, Dongwei Li, Ming Li
An efficient protocol to construct β-d-gluco-/galactosaminosyl linkages was established using nonparticipating and strong electron-withdrawing C-2-2,4-dinitrobenzenesulfonamide (DNsNH)-directed SN 2-like glycosylation of glycosyl ortho-hexynylbenzoates. The reaction is applicable to a wide range of O-, N-, and C-nucleophiles and features convenient conversion of DNsNH into AcNH in high yield under mild conditions. Oligomerization-ready trisaccharide, composed of β-d-(1→3)-glucosamino residues, has been achieved, setting a solid foundation for the synthesis of oligosaccharides associated with Neisseria meningitidis capsular polysaccharide...
March 22, 2019: Organic Letters
Ana Paula Rocha, Madalena Borges, Conceição Neves, João Farela Neves
We report a case of an 18-month-old boy with H factor deficiency with atypical presentation: recurrent acute otitis media and several maternal family members with autoimmune disorders (vitiligo, thyroiditis and immune trombocytopenia). Blood tests revealed low C3 and AH50, as well as low properdin and H factor. I factor was normal. CFH gene molecular test confirmed the H factor deficiency diagnosis. This child had none of the typical manifestations of this disorder, namely Neisseria meningitidis infection or renal disease (glomerulonephritis and atypical haemolytic uremic syndrome)...
February 28, 2019: Acta Médica Portuguesa
Bert Bogaerts, Raf Winand, Qiang Fu, Julien Van Braekel, Pieter-Jan Ceyssens, Wesley Mattheus, Sophie Bertrand, Sigrid C J De Keersmaecker, Nancy H C Roosens, Kevin Vanneste
Despite being a well-established research method, the use of whole-genome sequencing (WGS) for routine molecular typing and pathogen characterization remains a substantial challenge due to the required bioinformatics resources and/or expertise. Moreover, many national reference laboratories and centers, as well as other laboratories working under a quality system, require extensive validation to demonstrate that employed methods are "fit-for-purpose" and provide high-quality results. A harmonized framework with guidelines for the validation of WGS workflows does currently, however, not exist yet, despite several recent case studies highlighting the urgent need thereof...
2019: Frontiers in Microbiology
Wude Mihret, Berit Sletbakk Brusletto, Reidun Øvstebø, Anne-Marie Siebke Troseid, Gunnstein Norheim, Yared Merid, Afework Kassu, Workeabeba Abebe, Samuel Ayele, Mezgebu Silamsaw Asres, Lawrence Yamuah, Abraham Aseffa, Beyene Petros, Dominique A Caugant, Petter Brandtzaeg
Neisseria meningitidis infections in sub-Saharan Africa usually present with distinct symptoms of meningitis but very rarely as fulminant septicemia when reaching hospitals. In Europe, development of persistent meningococcal shock and multiple organ failure occurs in up to 30% of patients and is associated with a bacterial load of >106 /ml plasma or serum. We have prospectively studied 27 Ethiopian patients with meningococcal infection as diagnosed and quantified with real-time PCR in the cerebrospinal fluid (CSF) and serum...
April 2019: Innate Immunity
Sara Sigurlásdóttir, Gabriela M Wassing, Fanglei Zuo, Melanie Arts, Ann-Beth Jonsson
Neisseria meningitidis is a Gram-negative bacterium that asymptomatically colonizes the human nasopharyngeal mucosa. Pilus-mediated initial adherence of N. meningitidis to the epithelial mucosa is followed by the formation of three-dimensional aggregates, called microcolonies. Dispersal from microcolonies contributes to the transmission of N. meningitidis across the epithelial mucosa. We have recently discovered that environmental concentrations of host cell-derived lactate influences N. meningitidis microcolony dispersal...
2019: Frontiers in Microbiology
Enrique Chacon-Cruz, Christopher Roberts, Rosa Maria Rivas-Landeros, Erika Zoe Lopatynsky-Reyes, Lucila Alejandra Almada-Salazar, Jorge Arturo Alvelais-Palacios
Introduction: In Mexico, Neisseria meningitidis is considered to be a rare cause of bacterial meningitis (BM), however, one national publication using active surveillance has suggested the opposite. Group B Streptococcus (GBS) is also considered to be infrequent in young infants as a cause of BM in central Mexico. Streptococcus pneumoniae vaccination using the 13-valent conjugate vaccine (PCV13) started in our region in May 2012. We focused our research on whether N. meningitidis and GBS are important causes of BM, and to examine the effectiveness of PCV13 on pneumococcal BM...
January 2019: Therapeutic Advances in Infectious Disease
Matthias J H Gerritzen, Dirk E Martens, Joost P Uittenbogaard, René H Wijffels, Michiel Stork
Outer membrane vesicles (OMVs) produced by bacteria are interesting vaccine candidates. OMVs are nanoparticles that contain many immunogenic components, are self-adjuvating, and non-replicative. Despite recent insights in the biogenesis of OMVs, there is no consensus on a conserved mechanism of OMV release and the OMV yield from bacterial cultures remains low. For Neisseria meningitidis, a Gram-negative human pathogen causing meningitis and sepsis, a feasible OMV production method based on triggering OMV release by cysteine depletion has been described...
March 18, 2019: Scientific Reports
Caroline Cayrou, Christopher D Bayliss
Reverse transcriptase quantitative PCR (RT qPCR) is widely used for assessing the levels of expression of specific genes in various organisms. Here we describe a RT qPCR assay designed to determine the level of expression of fHbp in multiple isolates of Neisseria meningitidis. The level of expression is measured by a two-step qPCR and is associated with a promoter region analysis.
2019: Methods in Molecular Biology
Jakob Loschko, Karen Garcia, David Cooper, Michael Pride, Annaliesa Anderson
Flow cytometry provides an automated analysis of bacteria passing in fluid suspension through a laser light beam. Bacteria are first treated with antibodies that bind to a specific target. These antibodies are tagged to fluorophores that fluoresce when passed through a laser beam. As the bacteria pass sequentially through the laser beam, they absorb and scatter the light in forward and side (90°) angles. The forward angle scatter is proportional to the size of the bacteria and the 90° angle side scatter is proportional to the internal structure (granularity)...
2019: Methods in Molecular Biology
Elizabeth González, Fátima Reyes, Oscar Otero, Frank Camacho, Maribel Cuello, Fidel Ramírez, Reinaldo Acevedo
Vaccination has reduced morbidity and mortality of many diseases that previously caused devastating epidemics and deaths globally. Vaccines as a biological product may contain microorganisms or their derivatives. This aspect together with the fact that they are administered to healthy individuals (mainly children) means that approximately 70% of vaccines development time is dedicated to quality control. Monoclonal antibodies (MAbs) have become essential analytical tools for application in ELISAs, Western and Dot blotting, immunoprecipitation, and flow cytometric assays that ensure the quality control of vaccines...
2019: Methods in Molecular Biology
Jay Lucidarme, Jennifer Louth, Kelly Townsend-Payne, Ray Borrow
Serum bactericidal antibody (SBA) assays measure functional antibody titers against Neisseria meningitidis in sera. Induction of complement-dependent SBA after vaccination with meningococcal polysaccharide or conjugate or protein based vaccines is regarded as the surrogate of protection and thus acceptable evidence of the potential efficacy of these vaccines. This chapter discusses and details SBA assay protocols for measuring the complement-mediated lysis of serogroup A, B, C, W, X, and Y meningococci by human sera, for example, following vaccination or disease...
2019: Methods in Molecular Biology
Kay O Johswich, Scott D Gray-Owen
The single greatest barrier to studying the lifestyle of Neisseria meningitidis stems from its exquisite adaptation to life in humans, a specialization which prevents it from infecting other animals. This barrier to modeling meningococcal infection has been overcome by the provision of factors that allow the meningococci to overcome one or more aspects of host restriction, including the use of mice expressing receptors that allow mucosal colonization and/or the inclusion of serum factors that facilitate meningococcal replication during disseminated meningococcal disease...
2019: Methods in Molecular Biology
Brandon J Kim, Alexandra Schubert-Unkmeir
Bacterial meningitis is a serious, life-threatening infection of the central nervous system (CNS). To cause meningitis, bacteria must interact with and penetrate the meningeal blood-cerebrospinal fluid barrier (mB/CSFB), which comprises highly specialized brain endothelial cells. Neisseria meningitidis (meningococcus) is a leading cause of bacterial meningitis, and examination meningococcus' interaction with the BBB is critical for understanding disease progression. To examine specific interactions, in vitro mB/CSFB models have been developed and employed and are of great importance because in vivo models have been difficult to produce considering Neisseria meningitidis is exclusively a human pathogen...
2019: Methods in Molecular Biology
Emily A Kibble, Mitali Sarkar-Tyson, Geoffrey W Coombs, Charlene M Kahler
Neisseria meningitidis and Neisseria gonorrhoeae are obligate pathogens of the human host. Due to their adaptation to the human host, many factors required for infection are specialized for the human host to the point that natural infection processes are difficult to replicate in animal models. Immortalized human cell lines have been used to identify the host factors necessary for successful colonization of human mucosal surfaces. One such model is the Detroit 562 pharyngeal immortalized cell monolayer model which is used to measure the rate of attachment to and invasion of N...
2019: Methods in Molecular Biology
Tsitsi D Mubaiwa, Lauren E Hartley-Tassell, Evgeny A Semchenko, Christopher J Day, Michael P Jennings, Kate L Seib
A growing body of evidence suggests that glycans are important for meningococcal host-pathogen interactions and virulence. The development of glycobiology techniques such as glycan array analysis and surface plasmon resonance (SPR) has increased awareness of the importance of glycans in biological processes and has increased the interest of their study. While these techniques are more routinely used with purified proteins, there is growing interest in their applicability to cell-based studies, to better emulate host-pathogen interactions in vivo...
2019: Methods in Molecular Biology
Benjamin L Schulz
Mass spectrometry (MS) proteomics allows systematic identification, characterization, and relative quantification of the full suite of proteins in a biological sample, and is a powerful analytical approach for investigation of many aspects of the biology of Neisseria meningitidis. Here, we describe methods for robust and efficient sample preparation of the proteome of N. meningitidis suitable for diverse MS proteomics workflows.
2019: Methods in Molecular Biology
Freda E-C Jen, Kate L Seib, Aimee Tan
Fragment analysis (or fragment length analysis) is a PCR-based method which allows quantification of the size and proportion of a DNA repeat tract length of a phase-variable region. Primers are labeled with a fluorescent dye, the resulting amplicons are processed by capillary electrophoresis, and results are analyzed for amplicon size and proportion by associated software (such as Peakscanner). Here we describe the process of designing primers and controls to screen for the number of repeats in a polymeric tract of a phase-variable gene in Neisseria meningitidis (the DNA methyltransferase ModA is used as an example, but this method can be applied to other phase-variable genes)...
2019: Methods in Molecular Biology
Holly B Bratcher, Odile B Harrison, Martin C J Maiden
Whole genome sequencing (WGS) has revolutionized molecular microbiology, allowing the population biology of bacterial pathogens to be examined with greater accuracy and detail. The study of Neisseria meningitidis isolates, in particular, has benefitted from the availability of WGS data allowing outbreak cases, hyper-invasive lineages, molecular epidemiology, and vaccine coverage to be determined. Here, we describe a suite of protocols for the optimum recovery and analysis of WGS data, including a brief overview of methods for N...
2019: Methods in Molecular Biology
Nadja Heidrich, Saskia Bauriedl, Christoph Schoen
Deep sequencing technology has revolutionized transcriptome analyses of both prokaryotes and eukaryotes. RNA-sequencing (RNA-seq), which is based on massively parallel sequencing of cDNAs, has been used to annotate transcript boundaries and has revealed widespread antisense transcription as well as a wealth of novel noncoding transcripts in many bacterial pathogens. Moreover, RNA-seq is nowadays also widely used to comprehensively explore the interaction between RNA-binding proteins and their RNA targets on a genome-wide level in many human-pathogenic bacteria...
2019: Methods in Molecular Biology
Ala-Eddine Deghmane, Eva Hong, Muhamed-Kheir Taha
Neisseria meningitidis (Nm) is a leading cause of invasive infections associated with high mortality and morbidity, notably meningitis and septicemia. Etiological rapid diagnosis is key for the preventive management of invasive meningococcal disease (IMD). However, conventional methods for diagnosis are time-consuming and could be hampered by the difficulties in culturing the isolates from clinical specimens especially due to early antibiotic treatment. Therefore, sensitive, specific and rapid non-culture-based methods are valuable for early diagnosis, effective therapy, and prevention...
2019: Methods in Molecular Biology
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