Comparison of 16S ribosomal RNA gene sequence analysis and conventional culture in the environmental survey of hospital

学位記番号：医博甲160

Comparison of the efficacy of a commercial inactivated influenza A/H1N1/pdm09 virus (pH1N1) vaccine and two experimental M2e-based vaccines against pH1N1 challenge in the growing pig model

<div><p>Swine influenza A viruses (IAV-S) found in North American pigs are diverse and the lack of cross-protection among heterologous strains is a concern. The objective of this study was to compare a commercial inactivated A/H1N1/pdm09 (pH1N1) vaccine and two novel subunit vaccines, using IAV M2 ectodomain (M2e) epitopes as antigens, in a growing pig model. Thirty-nine 2-week-old IAV negative pigs were randomly assigned to five groups and rooms. At 3 weeks of age and again at 5 weeks of age, pigs were vaccinated intranasally with an experimental subunit particle vaccine (NvParticle/M2e) or a subunit complex-based vaccine (NvComplex/M2e) or intramuscularly with a commercial inactivated vaccine (Inact/pH1N1). At 7 weeks of age, the pigs were challenged with pH1N1 virus or sham-inoculated. Necropsy was conducted 5 days post pH1N1 challenge (dpc). At the time of challenge one of the Inact/pH1N1 pigs had seroconverted based on IAV nucleoprotein-based ELISA, Inact/pH1N1 pigs had significantly higher pdm09H1N1 hemagglutination inhibition (HI) titers compared to all other groups, and M2e-specific IgG responses were detected in the NvParticle/M2e and the NvComplex/M2e pigs with significantly higher group means in the NvComplex/M2e group compared to SHAMVAC-NEG pigs. After challenge, nasal IAV RNA shedding was significantly reduced in Inact/pH1N1 pigs compared to all other pH1N1 infected groups and this group also had reduced IAV RNA in oral fluids. The macroscopic lung lesions were characterized by mild-to-severe, multifocal-to-diffuse, cranioventral dark purple consolidated areas typical of IAV infection and were similar for NvParticle/M2e, NvComplex/M2e and SHAMVAC-IAV pigs. Lesions were significantly less severe in the SHAMVAC-NEG and the Inact/pH1N1pigs. Under the conditions of this study, a commercial Inact/pH1N1 specific vaccine effectively protected pigs against homologous challenge as evidenced by reduced clinical signs, virus shedding in nasal secretions and oral fluids and reduced macroscopic and microscopic lesions whereas intranasal vaccination with experimental M2e epitope-based subunit vaccines did not. The results further highlight the importance using IAV-S type specific vaccines in pigs.</p></div

A bibliometric analysis of m-learning from topic inception to 2015

Mobile learning is a promising and widely adopted mode of education nowadays. However, to the best of our knowledge, there have been no studies that have investigated publications on the use of mobile learning in the Thomson Reuters Web of Science (WoS) through bibliometric analysis. This study aims to provide readers with statistical information to obtain a deep understanding of this domain. The results show that 3087 mobile learning publications have been published since 1982. As a result, the study has compared and determined the possible ways for researchers and authors to improve the citation rate of their publications on m- learning and to gain a deeper understanding of the field of mobile learning by studying the number of publications and country, the number of authors, the keywords, the number of references, the number of pages, the journal, the authors' publications, and the number of citations. Additionally, other bibliometric results are discussed

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Not AvailableThe current inactivated influenza vaccines provide satisfactory protection against homologous viruses but limited cross-protection against antigenically divergent strains. Consequently, there is a need to develop more broadly protective vaccines. The highly conserved extracellular domain of the matrix protein 2 (M2e) has shown promising results as one of the components of a universal influenza vaccine in different animal models. As an approach to overcome the limited, strain specific, protective efficacy of inactivated influenza vaccine (IIV), a combination of recombinant M2e expressed on the surface of norovirus P particle (M2eP) and IIV was tested in chickens. Co-immunization of birds with both vaccines did not affect the production of M2e-specific IgG antibodies compared to the group vaccinated with M2eP alone. However, the co-immunized birds developed significantly higher pre-challenge hemagglutination inhibition antibody titers against the homologous IIV antigen and heterologous challenge virus. These combined vaccine groups also had cross reactive antibody responses against different viruses (H5, H6, and H7 subtypes) compared to the IIV alone vaccinated group. Upon intranasal challenge with homologous and heterologous viruses, the combined vaccine groups showed greater reduction in viral shedding in tracheal swabs compared to those groups receiving IIV alone. Moreover, M2eP antisera from vaccinated birds were able to bind to the native M2 expressed on the surface of whole virus particles and infected cells, and inhibit virus replication in vitro. Our results support the potential benefit of supplementing IIV with M2eP, to expand the vaccine cross protective efficacy.Not Availabl

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Not AvailablePigs are believed to be one of the important sources of emerging human and swine influenza viruses (SwIV). Influenza virus conserved peptides have the potential to elicit cross-protective immune response, but without the help of potent adjuvant and delivery system they are poorly immunogenic. Biodegradable polylactic-co-glycolic acid (PLGA) nanoparticle (PLGA-NP) based vaccine delivery system enhances cross-presentation of antigens by the professional antigen presenting cells. In this study, Norovirus P particle containing SwIV M2e (extracellular domain of the matrix protein 2) chimera and highly conserved two each of H1N1 peptides of pandemic 2009 and classical human influenza viruses were entrapped in PLGA-NPs. Influenza antibody-free pigs were vaccinated with PLGA-NPs peptides cocktail vaccine twice with or without an adjuvant, Mycobacterium vaccae whole cell lysate, intranasally as mist. Vaccinated pigs were challenged with a virulent heterologous zoonotic SwIV H1N1, and one week later euthanized and the lung samples were analyzed for the specific immune response and viral load. Clinically, pigs vaccinated with PLGA-NP peptides vaccine had no fever and flu symptoms, and the replicating challenged SwIV was undetectable in the bronchoalveolar lavage fluid. Immunologically, PLGA-NP peptides vaccination (without adjuvant) significantly increased the frequency of antigen-specific IFNγ secreting CD4 and CD8 T cells response in the lung lymphocytes, despite not boosting the antibody response both at pre- and post-challenge. In summary, our data indicated that nanoparticle-mediated delivery of conserved H1N1 influenza peptides induced the virus specific T cell response in the lungs and reduced the challenged heterologous virus load in the airways of pigsNot Availabl

The ISG and IFN responses in chickens vaccinated with LAIV or IIV at 1 day of age.

<p>(A) Trachea tissue at 1 dpv. (B) Trachea tissue at 3 dpv. (C) Spleen tissue at 1 dpv. (D) Spleen tissue at 3 dpv. The bars represent Log₂ fold change in transcription level of ISGs and IFN genes. Error bars represent mean±SD (n = 5 birds per group). Asterisks without horizontal bar indicate significant differences compared with unvaccinated group. Horizontal bar with asterisks indicates significant difference between LAIV and IIV (<i>*p<0</i>.<i>05</i>, <i>**p<0</i>.<i>01</i>). dpv = days post-vaccination.</p

Reduction in heterologous challenge virus replication in vaccinated birds.

<p>At 5 weeks of age (5 wpv for 1d IIV and 1d LAIV; 2 wpv for 3w LAIV and 3w IIV), chickens were challenged with heterologous virus and tracheal swabs were taken at 2 and 4 days post challenge to determine the level of challenge virus replication. Each swab was eluted in 2 ml of PBS. Virus titers are expressed as median egg infectious doses per ml of tracheal swab eluate. (A) 2 days post-challenge. (B) 4 days post-challenge. Groups are arranged as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0195285#pone.0195285.g005" target="_blank">Fig 5</a>. Different letters inside the plot indicate significant differences between groups (p<0.05).</p

Serum HI antibody response in chickens vaccinated at 1 day of age.

<p>(A) Homologous (TK/OR/71, H7N3) HI titers. (B) Heterologous (CK/NJ/02(H7N2)) HI titers. Sera were collected from birds vaccinated at 1 day of age at 14 days post-vaccination. Individual and median HI titers are indicated with symbols and horizontal lines, respectively. Different letters inside the plot indicate statistical significance among groups (p<0.05).</p