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  • 1
    Online Resource
    Online Resource
    Efficacy comparison of adjuvants in PcrV vaccine against Pseudomonas aeruginosa pneumonia
    Wiley ; 2017
    In:  Microbiology and Immunology Vol. 61, No. 2 ( 2017-02), p. 64-74
    Details
    In: Microbiology and Immunology, Wiley, Vol. 61, No. 2 ( 2017-02), p. 64-74
    Abstract: Vaccination against the type III secretion system of P. aeruginosa is a potential prophylactic strategy for reducing the incidence and improving the poor prognosis of P. aeruginosa pneumonia. In this study, the efficacies of three different adjuvants, Freund's adjuvant (FA), aluminum hydroxide (alum) and CpG oligodeoxynucleotide (ODN), were examined from the viewpoint of inducing PcrV‐specific immunity against virulent P. aeruginosa . Mice that had been immunized intraperitoneally with recombinant PcrV formulated with one of the above adjuvants were challenged intratracheally with a lethal dose of P. aeruginosa . The PcrV–FA immunized group attained a survival rate of 91%, whereas the survival rates of the PcrV–alum and PcrV–CpG groups were 73% and 64%, respectively. In terms of hypothermia recovery after bacterial instillation, PcrV–alum was the most protective, followed by PcrV–FA and PcrV–CpG. The lung edema index was lower in the PcrV–CpG vaccination group than in the other groups. PcrV–alum immunization was associated with the greatest decrease in myeloperoxidase in infected lungs, and also decreased the number of lung bacteria to a similar number as in the PcrV–FA group. There was less neutrophil recruitment in the lungs of mice vaccinated with PcrV–alum or PcrV–CpG than in those of mice vaccinated with PcrV–FA or PcrV alone. Overall, in terms of mouse survival the PcrV–CpG vaccine, which could be a relatively safe next‐generation vaccine, showed a comparable effect to the PcrV–alum vaccine.
    Type of Medium: Online Resource
    ISSN: 0385-5600 , 1348-0421
    URL: Issue
    URL: Article
    DOI: 10.1111/mim.v61.2
    DOI: 10.1111/1348-0421.12467
    Language: English
    Publisher: Wiley
    Publication Date: 2017
    detail.hit.zdb_id: 2102145-4
    SSG: 12
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  • 2
    Online Resource
    Online Resource
    Synthesis of 4′‐Thiomodified c‐di‐AMP Analogs
    Wiley ; 2023
    In:  Current Protocols Vol. 3, No. 9 ( 2023-09)
    Details
    In: Current Protocols, Wiley, Vol. 3, No. 9 ( 2023-09)
    Abstract: Cyclic diadenosine monophosphate (c‐di‐AMP) is a bacterial cyclic dinucleotide (CDN) comprising two adenosine monophosphates covalently linked by two 3′,5′‐phosphodiester bonds. c‐di‐AMP works as a second messenger, regulating many biological processes in bacteria such as cell wall homeostasis, DNA integrity, and sporulation via specific protein and/or RNA receptors. Moreover, c‐di‐AMP can function as an immunomodulatory agent in eukaryote cells via the stimulator of interferon genes (STING) signaling pathway. This protocol describes the chemical synthesis of two c‐di‐AMP analogs with a sulfur atom at the 4′‐position of the furanose ring instead of an oxygen atom: c‐di‐4′‐thioAMP ( 1 ) and cAMP‐4′‐thioAMP ( 2 ). Analogs 1 and 2 have resistance to phosphodiesterase‐mediated degradation and are therefore useful for understanding the diverse biological phenomena regulated by c‐di‐AMP. In this protocol, two 4′‐thioadenosine monomers are initially prepared via a Pummerer‐like reaction assisted by hypervalent iodine. The CDN skeleton is then constructed through two key reactions based on phosphoramidite chemistry: dimerization of two appropriately protected nucleoside monomers to produce a linear dinucleotide, followed by macrocyclization of the resulting linear dinucleotide to form the CDN skeleton. © 2023 Wiley Periodicals LLC. Basic Protocol 1 : Preparation of 4′‐thioadenosine monomers 13 and 14 Basic Protocol 2 : Preparation of c‐di‐4′‐thioAMP ( 1 ) and cAMP‐4′‐thioAMP ( 2 )
    Type of Medium: Online Resource
    ISSN: 2691-1299 , 2691-1299
    URL: Issue
    URL: Article
    DOI: 10.1002/cpz1.v3.9
    DOI: 10.1002/cpz1.892
    Language: English
    Publisher: Wiley
    Publication Date: 2023
    detail.hit.zdb_id: 3059383-9
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  • 3
    Online Resource
    Online Resource
    Epidemiological analysis of serum anti‐ Pseudomonas aeruginosa PcrV titers in adults
    Wiley ; 2016
    In:  Microbiology and Immunology Vol. 60, No. 2 ( 2016-02), p. 114-120
    Details
    In: Microbiology and Immunology, Wiley, Vol. 60, No. 2 ( 2016-02), p. 114-120
    Abstract: Of the various virulence mechanisms of the opportunistic pathogen Pseudomonas aeruginosa , the type III secretion system (TTSS) has been characterized as a major factor associated with acute lung injury, bacteremia and mortality. In addition, PcrV, a component protein of the TTSS, has been characterized as a protective antigen against infection with P. aeruginosa . This study comprised an epidemiological analysis of serum anti‐PcrV titers in a cohort of Japanese adults. From April 2012 to March 2013, serum anti‐PcrV titers of 198 volunteer participants undergoing anesthesia for scheduled surgeries were measured. The median, minimum and maximum serum anti‐PcrV titers among the 198 participants were 4.09 nM, 1.01 nM and 113.81 nM, respectively. The maximum peaks in the histogram were within the anti‐PcrV 2.00–4.99 nM titer range; values for 115 participants (58.1%) were within this range. Anti‐PcrV titers were more than approximately three‐fold greater ( 〉 12 nM) than the median value in 21 participants (10.6%). Ten‐year interval age increases, history of treatment for traffic trauma, and a history of past surgery each showed statistically significant associations with higher anti‐PcrV titers (i.e., 〉 10 nM) than did the other factors assessed by binomial analysis. This study revealed a considerable variation in anti‐PcrV titers in adult subjects without any obvious histories of infection with P. aeruginosa .
    Type of Medium: Online Resource
    ISSN: 0385-5600 , 1348-0421
    URL: Issue
    URL: Article
    DOI: 10.1111/mim.v60.2
    DOI: 10.1111/1348-0421.12353
    Language: English
    Publisher: Wiley
    Publication Date: 2016
    detail.hit.zdb_id: 2102145-4
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    The efficacy of basic fibroblast growth factor‐loaded poly(lactic‐ co ‐glycolic acid) nanosheet for mouse wound healing
    Wiley ; 2017
    In:  Wound Repair and Regeneration Vol. 25, No. 6 ( 2017-11), p. 1008-1016
    Details
    In: Wound Repair and Regeneration, Wiley, Vol. 25, No. 6 ( 2017-11), p. 1008-1016
    Abstract: Although human recombinant basic fibroblast growth factor (bFGF) is widely used for wound healing, daily treatment with bFGF is required because of its short half‐life. An effective controlled‐release system of bFGF is, therefore, desired in clinical settings. To investigate the efficacy of a bFGF‐loaded nanosheet for wound healing, focusing on the controlled‐release of bFGF, bFGF‐loaded poly(lactic‐ co ‐glycolic acid) (PGLA) nanosheets were developed, and their in vitro release profile of bFGF and their in vivo efficacy for wound healing were examined. A polyion complex of positively charged human recombinant bFGF and negatively charged alginate was sandwiched between PLGA nanosheets (70 nm thick for each layer). The resulting bFGF‐loaded nanosheet robustly adhered to silicon skin by observation using a microscratch test. bFGF was gradually and continuously released over three days in an in vitro incubation study. Treatment with the bFGF‐loaded nanosheets (every 3 day for 15 days) as well as with a conventional bFGF spray effectively promoted wound healing of mouse dorsal skin defects with accelerated tissue granulation and angiogenesis, although the dose of bFGF used in the treatment with the bFGF nanosheets was approximately 1/20 of the sprayed bFGF. In conclusion, we developed a bFGF‐loaded nanosheet that sustained a continuous release of bFGF over three days and effectively promoted wound healing in mice.
    Type of Medium: Online Resource
    ISSN: 1067-1927 , 1524-475X
    URL: Issue
    URL: Article
    DOI: 10.1111/wrr.2017.25.issue-6
    DOI: 10.1111/wrr.12604
    Language: English
    Publisher: Wiley
    Publication Date: 2017
    detail.hit.zdb_id: 2011990-2
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  • 5
    Online Resource
    Online Resource
    The protective effects of nasal PcrV‐CpG oligonucleotide vaccination against Pseudomonas aeruginosa pneumonia
    Wiley ; 2018
    In:  Microbiology and Immunology Vol. 62, No. 12 ( 2018-12), p. 774-785
    Details
    In: Microbiology and Immunology, Wiley, Vol. 62, No. 12 ( 2018-12), p. 774-785
    Abstract: An effective vaccine against Pseudomonas aeruginosa would be hugely beneficial to people who are susceptible to the serious infections it can cause. Vaccination against PcrV of the P. aeruginosa type III secretion system is a potential prophylactic strategy for improving the incidence and prognosis of P. aeruginosa pneumonia. Here, the effect of nasal PcrV adjuvanted with CpG oligodeoxynucleotide (CpG) was compared with a nasal PcrV/aluminum hydroxide gel (alum) vaccine. Seven groups of mice were vaccinated intranasally with one of the following: 1, PcrV‐CpG; 2, PcrV‐alum; 3, PcrV alone; 4, CpG alone; 5, alum alone; 6 and 7, saline control. Fifty days after the first immunization, anti‐PcrV IgG, IgA and IgG isotype titers were measured; significant increases in these titers were detected only in the PcrV‐CpG vaccinated mice. The vaccinated mice were then intratracheally infected with a lethal dose of P. aeruginosa and their body temperatures and survival monitored for 24 hr, edema, bacteria, myeloperoxidase activity and lung histology also being evaluated at 24 hr post‐infection. It was found that 73% of the PcrV‐CpG‐vaccinated mice survived, whereas fewer than 30% of the mice vaccinated with PcrV‐alum or adjuvant alone survived. Lung edema and other inflammation‐related variables were less severe in the PcrV‐CpG group. The significant increase in PcrV‐specific IgA titers detected following PcrV‐CpG vaccination is probably a component of the disease protection mechanism. Overall, our data show that intranasal PcrV‐CpG vaccination has potential efficacy for clinical application against P. aeruginosa pneumonia.
    Type of Medium: Online Resource
    ISSN: 0385-5600 , 1348-0421
    URL: Issue
    URL: Article
    DOI: 10.1111/mim.v62.12
    DOI: 10.1111/1348-0421.12658
    Language: English
    Publisher: Wiley
    Publication Date: 2018
    detail.hit.zdb_id: 2102145-4
    SSG: 12
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  • 6
    Online Resource
    Online Resource
    Molecular epidemiology of clinically high‐risk Pseudomonas aeruginosa strains: Practical overview
    Wiley ; 2020
    In:  Microbiology and Immunology Vol. 64, No. 5 ( 2020-05), p. 331-344
    Details
    In: Microbiology and Immunology, Wiley, Vol. 64, No. 5 ( 2020-05), p. 331-344
    Abstract: In recent years, numerous outbreaks of multidrug‐resistant Pseudomonas aeruginosa have been reported across the world. Once an outbreak occurs, besides routinely testing isolates for susceptibility to antimicrobials, it is required to check their virulence genotypes and clonality profiles. Replacing pulsed‐field gel electrophoresis DNA fingerprinting are faster, easier‐to‐use, and less expensive polymerase chain reaction (PCR)‐based methods for characterizing hospital isolates. P. aeruginosa possesses a mosaic genome structure and a highly conserved core genome displaying low sequence diversity and a highly variable accessory genome that communicates with other Pseudomonas species via horizontal gene transfer. Multiple‐locus variable‐number tandem‐repeat analysis and multilocus sequence typing methods allow for phylogenetic analysis of isolates by PCR amplification of target genes with the support of Internet‐based services. The target genes located in the core genome regions usually contain low‐frequency mutations, allowing the resulting phylogenetic trees to infer evolutionary processes. The multiplex PCR‐based open reading frame typing (POT) method, integron PCR, and exoenzyme genotyping can determine a genotype by PCR amplifying a specific insertion gene in the accessory genome region using a single or a multiple primer set. Thus, analyzing P. aeruginosa isolates for their clonality, virulence factors, and resistance characteristics is achievable by combining the clonality evaluation of the core genome based on multiple‐locus targeting methods with other methods that can identify specific virulence and antimicrobial genes. Software packages such as eBURST, R, and Dendroscope, which are powerful tools for phylogenetic analyses, enable researchers and clinicians to visualize clonality associations in clinical isolates.
    Type of Medium: Online Resource
    ISSN: 0385-5600 , 1348-0421
    URL: Issue
    URL: Article
    DOI: 10.1111/mim.v64.5
    DOI: 10.1111/1348-0421.12776
    Language: English
    Publisher: Wiley
    Publication Date: 2020
    detail.hit.zdb_id: 2102145-4
    SSG: 12
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