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Current Protocols in Microbiology

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https://www.readbyqxmd.com/read/30016567/genetic-manipulation-of-cryptococcus-neoformans
#1
Kwang-Woo Jung, Kyung-Tae Lee, Yee-Seul So, Yong-Sun Bahn
Cryptococcus neoformans is an opportunistic fungal pathogen, which causes life-threatening meningoencephalitis in immunocompromised individuals and is responsible for more than 1,000,000 infections and 600,000 deaths annually worldwide. Nevertheless, anti-cryptococcal therapeutic options are limited, mainly because of the similarity between fungal and human cellular structures. Owing to advances in genetic and molecular techniques and bioinformatics in the past decade, C. neoformans, belonging to the phylum basidiomycota, is now a major pathogenic fungal model system...
July 17, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29995344/in-vitro-culturing-and-screening-of-candida-albicans-biofilms
#2
Megha Gulati, Matthew B Lohse, Craig L Ennis, Ruth E Gonzalez, Austin M Perry, Priyanka Bapat, Ashley Valle Arevalo, Diana L Rodriguez, Clarissa J Nobile
Candida albicans is a normal member of the human microbiota that asymptomatically colonizes healthy individuals, however it is also an opportunistic pathogen that can cause severe infections, especially in immunocompromised individuals. The medical impact of C. albicans depends, in part, on its ability to form biofilms, communities of adhered cells encased in an extracellular matrix. Biofilms can form on both biotic and abiotic surfaces, such as tissues and implanted medical devices. Once formed, biofilms are highly resistant to antifungal agents and the host immune system, and can act as a protected reservoir to seed disseminated infections...
July 11, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29927109/shigella-pathogenesis-modeling-with-tissue-culture-assays
#3
Benjamin J Koestler, Cara M Ward, Shelley M Payne
Shigella is an enteroinvasive human pathogen that infects the colonic epithelium and causes Shigellosis, an infectious diarrheal disease. There is no vaccine for the prevention or treatment of Shigellosis and antibiotic-resistant strains of Shigella are increasing, emphasizing the need for a deeper understanding of Shigella pathogenesis in order to design effective antimicrobial therapies. Small animal models do not recapitulate Shigellosis, therefore tissue-cultured cells have served as model systems to study Shigella pathogenesis...
May 24, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29927105/use-of-axenic-culture-tools-to-study-coxiella-burnetii
#4
Savannah E Sanchez, Eduardo Vallejo-Esquerra, Anders Omsland
Coxiella burnetii is a highly infectious obligate intracellular bacterium and the etiological agent of the zoonosis Query (Q) fever. This Gram-negative gamma-proteobacterium has adapted to replicate within a specialized compartment in mammalian phagocytic cells, known as the Coxiella-containing vacuole (CCV). Knowledge of critical characteristics of the CCV microenvironment (e.g., luminal pH), analysis of the C. burnetii genome sequence, and strategic metabolic profiling have provided the basis for determining the physicochemical and nutritional conditions necessary to support axenic replication of C...
May 18, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29927091/salmonella-typhimurium-infection-of-human-monocyte-derived-macrophages
#5
Stephanie K Lathrop, Kendal G Cooper, Kelsey A Binder, Tregei Starr, Veena Mampilli, Corrella S Detweiler, Olivia Steele-Mortimer
The successful infection of macrophages by non-typhoidal serovars of Salmonella enterica is likely essential to the establishment of the systemic disease they sometimes cause in susceptible human populations. However, the interactions between Salmonella and human macrophages are not widely studied, with mouse macrophages being a much more common model system. Fundamental differences between mouse and human macrophages make this less than ideal. Additionally, the inability of human macrophage-like cell lines to replicate some properties of primary macrophages makes the use of primary cells desirable...
May 18, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29927096/human-intestinal-enteroids-for-the-study-of-bacterial-adherence-invasion-and-translocation
#6
Nina M Poole, Anubama Rajan, Anthony W Maresso
Adherence, invasion, and translocation to and through the intestinal epithelium are important drivers of disease for many enteric bacteria. However, most work has been limited to transformed intestinal cell lines or murine models that often do not faithfully recapitulate key elements associated with human disease. The recent technological advances in organotypic tissue and cell culture are providing unparalleled access to systems with human physiology and complexity. Human intestinal enteroids (HIEs), derived from patient biopsy or surgical specimens of intestinal tissues, are organotypic cultures now being adapted to the study of enteric infections...
May 17, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512119/reverse-genetics-for-newcastle-disease-virus-as-a-vaccine-vector
#7
Shin-Hee Kim, Siba K Samal
Newcastle disease virus (NDV) is an economically important pathogen in the poultry industry worldwide. Recovery of infectious NDV from cDNA using reverse genetics has made it possible to manipulate the genome of NDV. This has greatly contributed to our understanding of the molecular biology and pathogenesis of NDV. Furthermore, NDV has modular genome and accommodates insertion of a foreign gene as a transcriptional unit, thus enabling NDV as a vaccine vector against diseases of humans and animals. Avirulent NDV strains (e...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512118/analyzing-chemotaxis-and-related-behaviors-of-azospirillum-brasilense
#8
Lindsey O'Neal, Tanmoy Mukherjee, Gladys Alexandre
Bacteria of the genus A. brasilense are motile and capable of chemotaxis and aerotaxis (taxis in gradient of oxygen) using a single polar flagellum that propels the cells in aqueous environments. Responses to attractants and repellents have been described and spatial gradient assays that permit the visualization of these responses are detailed in this unit. These assays are simple and can be readily implemented with minimum set ups. © 2018 by John Wiley & Sons, Inc.
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512117/growth-and-laboratory-maintenance-of-paenibacillus-larvae
#9
Osama S Mahdi, Nathan A Fisher
Paenibacillus larvae is a Gram-positive, spore-forming bacterium and the causative agent of American foulbrood disease (AFB), a highly contagious, fatal disease affecting managed honeybee (Apis mellifera) colonies. As the etiological agent of American foulbrood disease, P. larvae is the most economically significant bacterial pathogen infecting honeybees. This unit includes protocols for the in vitro growth and laboratory maintenance of P. larvae. © 2018 by John Wiley & Sons, Inc.
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512116/sporulation-and-germination-of-paenibacillus-larvae-cells
#10
Osama S Mahdi, Nathan A Fisher
Endospores are metabolically dormant cells formed by a variety of Gram-positive bacteria within the phylum Firmicutes in response to nutrient limiting or otherwise unfavorable growth conditions. American foulbrood disease (AFB) is a serious disease of honeybees that is caused by the introduction of Paenibacillus larvae endospores into a honeybee colony. Progression to fulminant disease and eventual collapse of the colony requires multiple rounds of endospore germination, vegetative replication, endospore formation, and subsequent spread within the colony...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512115/hepatitis-e-virus-isolation-propagation-and-quantification
#11
Shaoli Lin, Liping Yang, Yan-Jin Zhang
Hepatitis E virus (HEV) predominantly causes acute liver disease in humans and is transmitted via the fecal-oral route. HEV infection in pregnant women can result in grave consequences, with up to 30% fatality. The HEV strains infecting humans mainly belong to four genotypes. Genotypes 1 and 2 are restricted to human infection, while genotypes 3 and 4 are zoonotic. HEV genotype 3 (HEV-3) can cause both acute and chronic liver diseases. Several cell lines (mainly hepatocytes) have been developed for HEV propagation and biological study...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512114/poliovirus-replicon-rna-generation-transfection-packaging-and-quantitation-of-replication
#12
Ekaterina G Viktorova, Sunil Khattar, Siba Samal, George A Belov
Poliovirus is a prototype member of the Enterovirus genus of the Picornaviridae family of small positive strand RNA viruses, which include important human and animal pathogens. Quantitative assessment of viral replication is very important for investigation of the virus biology and the development of anti-viral strategies. The poliovirus genome structure allows replacement of structural genes with a reporter protein, such as a luciferase or a fluorescent protein, whose signals can be detected and quantified in vivo, thus permitting observation of replication kinetics in live cells...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512113/porcine-reproductive-and-respiratory-syndrome-virus-propagation-and-quantification
#13
Zexu Ma, Liping Yang, Yan-Jin Zhang
Porcine reproductive and respiratory syndrome virus (PRRSV) is a member of the family Arteriviridae, order Nidovirale. PRRSV is an enveloped, single-stranded, positive-sense RNA virus with a genome around 15 kb in length. For propagation of PRRSV in vitro, the MARC-145 cell line is the most often used in a laboratory setting. Infectious cDNA clones of many PRRSV strains have been established, from which these viruses can be recovered. PRRSV titration is generally done in MARC-145 cells. PRRSV RNA copy numbers can be assessed by reverse transcription and real-time PCR...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29512112/molecular-genetic-manipulation-of-sarcocystis-neurona
#14
Daniel K Howe, Michelle Yeargan, Landon Simpson, Sriveny Dangoudoubiyam
Sarcocystis neurona is a member of the important phylum Apicomplexa and the primary cause of equine protozoal myeloencephalitis (EPM). Moreover, S. neurona is the best-studied species in the genus Sarcocystis, one of the most successful parasite taxa, as virtually all vertebrate animals may be infected by at least one species. Consequently, scientific investigation of S. neurona will aid in the control of EPM and neurologic disease in sea mammals, while also improving our understanding of a prominent branch on the apicomplexan phylogenetic tree...
February 22, 2018: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120487/azospirillum-brasilense-a-beneficial-soil-bacterium-isolation-and-cultivation
#15
Gladys Alexandre
Bacteria of the genus Azospirillum comprise 15 species to date, with A. brasilense the best studied species in the genus. Azospirillum are soil bacteria able to promote the growth of plants from 113 species spanning 35 botanical families. These non-pathogenic and beneficial bacteria are ubiquitous in soils and inhabit the roots of diverse plants. These bacteria are microaerophilic, able to fix nitrogen under free-living conditions, motile, and able to navigate in gradients of various chemicals, including oxygen...
November 9, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120486/computational-methods-for-human-microbiome-analysis
#16
Matthieu J Miossec, Sandro L Valenzuela, Katterinne N Mendez, Eduardo Castro-Nallar
As the field of microbiomics advances, the burden of computational work that scientists need to perform in order to extract biological insight has grown accordingly. Likewise, while human microbiome analyses are increasingly shifting toward a greater integration of various high-throughput data types, a core number of methods form the basis of nearly every study. In this unit, we present step-by-step protocols for five core stages of human microbiome research. The protocols presented in this unit provide a base case for human microbiome analysis, complete with sufficient detail for researchers to tailor certain aspects of the protocols to the specificities of their data...
November 9, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120485/measuring-ph-of-the-coxiella-burnetii-parasitophorous-vacuole
#17
Dhritiman Samanta, Stacey D Gilk
Coxiella burnetii is the causative agent of human Q fever, a zoonotic disease that can cause a debilitating, flu-like illness in acute cases, or a life-threatening endocarditis in chronic patients. An obligate intracellular bacterial pathogen, Coxiella survives and multiplies in a large lysosome-like vacuole known as the Coxiella parasitophorous vacuole (CPV). A unique characteristic of the CPV is the acidic environment (pH ∼5.0), which is required to activate Coxiella metabolism and the Coxiella type 4 secretion system (T4SS), a major virulence factor required for intracellular survival...
November 9, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120484/elisa-for-molluscum-contagiosum-virus
#18
Subuhi Sherwani, Mohammed Chowdhury, Joachim J Bugert
Molluscum contagiosum virus (MCV) is a common skin pathogen of children and young adults. Infection with MCV causes benign skin tumors in children and young adults and is mostly self-limiting. In contrast to orthopoxviruses, MCV infections tend to take a subacute clinical course but may persist for up to 12 months. Current numbers for MCV seroprevalence in different geographical areas are based on a variety of historical serological methods from complement fixation assays to MCV ELISAs based on purified MCV virions and MC133 antigen expressed in a Semliki Forest Virus expression system...
November 9, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120483/azospirillum-brasilense-laboratory-maintenance-and-genetic-manipulation
#19
Jessica Gullett, Lindsey O'Neal, Tanmoy Mukherjee, Gladys Alexandre
Bacteria of the genus Azospirillum, including the most comprehensively studied Azospirillum brasilense, are non-pathogenic soil bacteria that promote the growth of diverse plants, making them an attractive model to understand non-symbiotic, beneficial plant-bacteria associations. Research into the physiology and genetics of these organisms spans decades and a range of molecular tools and protocols have been developed for allelic exchange mutagenesis, in trans expression of genes, and fusions to reporter genes...
November 9, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/29120482/leptospira-molecular-detection-of-pathogenic-species-in-natural-sources
#20
Brittney Beigel, Ashutosh Verma
This protocol describes a method for the rapid detection of leptospiral DNA in environmental water. In summary, the DNA is extracted from water samples and tested in a TaqMan-based real-time quantitative polymerase chain reaction (qPCR) for the presence of lipl32, a gene that is present only in pathogenic Leptospira spp. The gene target used in this assay is important in that it only detects pathogenic leptospires and not the saprophytic leptospires that may be present in environmental samples. © 2017 by John Wiley & Sons, Inc...
November 9, 2017: Current Protocols in Microbiology
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