Expression and purification of pneumococcal surface protein a of clade 4 in Escherichia coli using hydroxylapatite and ion-exchange column chromatography
In this study, we describe a simple method to produce PspA of clade 4 from an Escherichia coli expression system using hydroxylapatite and ion-exchange chromatography. Using this method, we successfully expressed soluble PspA4 in 10 L of autoinducing culture medium, with a wet-cell yield of 19 g/L and a final PspA4 concentration of 22.8 mg/L. Additionally, we improved PspA4 purity from 17% to 70% in a single step through the use of hydroxylapatite, resulting in acquisition of recombinant PspA4 (>95% purity) at a final yield of 43% from the starting cell-lysis solution. We subsequently verified the secondary structure molecular weight of recombinant PspA4 by circular dichroism and mass spectrometry, respectively. These results demonstrated a highly efficient method for mass producing PspA4 protein and that can also be applied for purification of PspA proteins from other clades.
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CONCLUSION: BPPV is the most frequent diagnosis seen in the ED; however, physicians need to document nystagmus more precisely and perform diagnostic tests systematically, in order to make an accurate diagnosis. To avoid misdiagnoses, ED physicians and ENT specialists should be able to recognize central signs in patients with an acute vestibular syndrome. Every fourth patient does not receive a definitive diagnosis. Diagnostic ED workup for patients with dizziness needs further improvement. PMID: 32221628 [PubMed - as supplied by publisher]
AbstractCytomegalovirus (CMV) infection causes significant morbidity and mortality in immunocompromised transplant patients. ASP0113, a first ‐in‐class DNA vaccine containing plasmids encoding CMV phosphoprotein 65 and glycoprotein B (gB), was evaluated in a phase 1b, subject‐blinded study in CMV‐seropositive (n = 13) and CMV‐seronegative (n = 12) healthy and CMV‐seronegative dialysis subjects (n = 12) randomized to ASP0 113 or placebo. End points included pharmacokinetics, anti‐gB antibody levels, phosphoprotein 65–specific T‐cell responses measured by ex vivo enzyme‐l...
ConclusionsThus, hMCs exhibit a highly individualized pattern of immune response possibly to meet tissue requirements and regulate bacteria coexistence vs defense.
Publication date: Available online 30 March 2020Source: Mutation Research/Fundamental and Molecular Mechanisms of MutagenesisAuthor(s): Kristen Fernandez, Sara D’Souza, Jenny J. Ahn, Summer Singh, Erin Mae Bacasen, Daniel Mashiach, Daniel Mishail, Timothy Kao, Jasmine Thai, Spring Hwang, Lekha Yaramada, Jeffrey H. Miller
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Publication date: Available online 30 March 2020Source: LWTAuthor(s): David R. Kasler, Taras Pyatkovskyy, Ahmed E. Yousef, Sudhir K. Sastry
Biofilms usually occur on or within medical devices such as mechanical heart valves, prosthetic joints, peritoneal dialysis catheters, contact lenses, central venous catheters, pacemakers and urinary catheters. The presence of biofilms can be seen in different diseases like endocarditis, otitis, cystic fibrosis, periodontitis, osteomyelitis and chronic wounds. Toxin–antitoxin systems are reported to respond to stresses that are also found in the host. Some toxins of toxin–antitoxin systems modules have been proposed as potential weapons used by bacteria to act on host targets. In this review, we focus on the mo...
The antimicrobial susceptibility testing of 284 Enterobacteriaceae isolates responsible for urinary tract infections to ampicillin, ceftazidime, ciprofloxacin, nitrofurantoin, trimethoprin-sulfamethaxole, and fosfomycin was performed by disk diffusion method. Additionally, in fosfomycin-resistant and intermediate susceptible isolates using disk diffusion method, the minimum inhibitory concentration (MIC) of fosfomycin was determined by agar dilution. The presence of fosA and fosA3 genes and ESBL genes was investigated in fosfomycin-resistant isolates and ESBL-producing isolates, respectively. Klebsiella pneumoniae [72.34% ...