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Microbiology and Immunology

Irina N Protasova, Tsai-Wen Wan, Natalya V Bakhareva, Wei-Chun Hung, Wataru Higuchi, Yasuhisa Iwao, Tatyana A Yelistratova, Natalya A Ilyenkova, Yelena S Sokolovskaya, Galina P Martynova, Ivan V Reva, Galina V Reva, Sergey V Sidorenko, Lee-Jene Teng, Olga V Peryanova, Alla B Salmina, Tatsuo Yamamoto
Streptococcus pneumoniae, a common human pathogen, colonizes the nasopharynx and causes diseases including acute otitis media (AOM). We herein report pneumococcal serotype distributions in children before and after PCV7 vaccination and in patients with pneumococcal disease in Siberian Russia (Krasnoyarsk). Analyses included antimicrobial susceptibility testing, ST typing, a pulsed-field gel electrophoresis analysis, virulence-related surface protein gene (VSG) typing with novel primers, and a structural analysis by scanning electron microscopy...
July 23, 2017: Microbiology and Immunology
Si-Biao Su, Jian-Feng Zhang, Fei-Fei Huang, Yu Cen, Hai-Xing Jiang
Cholangiocarcinoma (CCA) associated with liver fluke infection appears to involve inflammatory and immune processes, but whether these involve the proinflammatory cytokine interleukin-17A (IL-17A) and proliferative cytokine interleukin-22 (IL-22) is unclear.We compared levels of IL-22- or IL-17A-producing T helper cells and cytokine levels in 30 CCA patients with long-term liver fluke infection, 40 patients with liver-fluke infection but not CCA and 16 healthy controls. Analyses were performed using immunohistochemistry, flow cytometry, ELISA and RT-PCR...
July 18, 2017: Microbiology and Immunology
Yuki Matsushima, Tomomi Shimizu, Ikuko Doi, Fuminori Mizukoshi, Koo Nagasawa, Akihide Ryo, Hideaki Shimizu, Masae Kobayashi, Keiji Funatogawa, Noriko Nagata, Mariko Ishikawa, Ayako Komane, Nobuhiko Okabe, Yoshio Mori, Makoto Takeda, Hirokazu Kimura
To detect various viral genes in the patients with rash and fever illness (RFI), a new multiplex (reverse transcription) polymerase chain reaction [(RT)-PCR] method was constructed. New primer sets were designed for the detection of herpes simplex viruses 1 and 2 (HSV1 and 2), and Epstein-Barr virus (EBV). The newly designed and previously reported primer sets were used to detect 13 types of RFI-associated viruses by multiplex (RT)-PCR assay systems. Moreover, to eliminate non-specific PCR products, a double-stranded specific DNase was used for the digestion of double-stranded DNA derived from the templates in clinical specimens...
July 15, 2017: Microbiology and Immunology
Chantal Ama Agbemabiese, Toyoko Nakagomi, Minh Quang Nguyen, Punita Gauchan, Osamu Nakagomi
One major mechanism by which Rotavirus A (RVA) evolves is genetic reassortment between strains possessing different genotype constellations. The parental strains of the reassortants generated are however seldom identified. Here, we examined the whole genome of two suspected reassortants RVA/Human-wt/VNM/SP127/2013/G1P[4] and RVA/Human-wt/VNM/SP193/2013/G1P[4] with short RNA electropherotypes by Illumina MiSeq sequencing and reconstructed their ancestral phylogenies. Their genotype constellation - G1-P[4]-I2-R2-C2-M2-A2-N2-T2-E2-H2 indicated that they were G1 VP7 mono-reassortants possessing DS-1-like genetic backbones...
July 11, 2017: Microbiology and Immunology
Yothin Teethaisong, Katie Evans, Ismini Nakouti, Kanokwan Tiamyom, James R Ketudat-Cairns, Glyn Hobbs, Griangsak Eumkeb
A resazurin chromogenic agar (RCA) along with combined disc method has been developed as a promising method for rapid screening of extended-spectrum-β-lactamase (ESBL), AmpC β-lactamase, and co-production of ESBL and AmpC. Cefpodoxime (CPD) discs supplemented with and without clavulanic acid (CA), cloxacillin (CX), or CA+CX were evaluated against 86-molecularly confirmed β-lactamase-producing Enterobacteriaceae, including 15 ESBLs, 32 AmpCs, 9 co-producers of ESBL and AmpC, and 30 carbapenemase producers...
July 7, 2017: Microbiology and Immunology
Tat-Chuan Cham, Yen-Chen Chang, Pei-Shiue Tsai, Ching-Ho Wu, Hui-Wen Chen, Chian-Ren Jeng, Victor Fei Pang, Hui-Wen Chang
Unlike serotype II feline coronaviruses (FCoV II), the cellular receptor for serotype I FCoV (FCoV I), the most prevalence FCoV serotype, is unknown. To provide a platform to assess attachment pattern of the FCoV I with its host receptor(s), we established HEK293 cell lines that stably expressed the ectodomains of the spike (S) proteins derived from a FCoV I feline enteric coronavirus strain UU7 (FECV UU7) and a feline infectious peritonitis virus strain UU4 (FIPV UU4). Using the recombinant S proteins as probes to perform S protein affinity histochemistry in paraffin-embedded tissues, despite no tissue or enteric binding of FECV UU7 S protein was detected, we showed that the tissue distribution of FIPV UU4 S protein-bound cells correlated with that of FIPV antigen-positive cells by immunohistochemistry and lesions associated with FIP; and the affinity binding of FIPV UU4 S protein on macrophages was not affected by the enzymatic removal of host cell-surface sialic acid with neuraminidase...
July 4, 2017: Microbiology and Immunology
Nattika Nantachit, Panya Sunintaboon, Sukathida Ubol
Domain III of E protein of DENV is one of the targets for vaccine development. Unfortunately, this protein-based platform has generally low immunogenicity. To circumvent this problem, we investigated the use of an adjuvant-nanoparticle delivery system to facilitate immunogenicity of soluble DENV-EDIII protein. One of the key features of this delivery system is its ability to simultaneously deliver antigens and exert adjuvanticity to specialized immune cells. In this study, we generated N-trimethyl chitosan (TMC) nanoparticles (NPs) to be used as adjuvant and carrier for the soluble E-domain III of dengue virus serotype 3 (sEDIII-D3)...
June 12, 2017: Microbiology and Immunology
Toyotaka Sato, Hidetoshi Higuchi, Shin-Ichi Yokota, Yutaka Tamura
Erythromycin, tylosin, and tilmicosin are approved for use in cattle in Japan, and later two are used for treatment of Mycoplasma bovis infection. In this study, 58 M. bovis isolates obtained from Japanese dairy calves all exhibited reduced susceptibility to these macrolides. The widespread reduced susceptibility to the macrolides resulted from a few dominant lineages. All 58 isolates contained the G748A variant in both the rrl3 and rrl4 alleles of 23S rRNA, whereas a reference strain (PG45) did not. G748 localizes in the central loop of domain II (from C744 to A753) of 23S rRNA that participates in binding to mycinose, a sugar residue present in both tylosin and tilmicosin...
May 15, 2017: Microbiology and Immunology
Yan-Mei Jiao, Guang-Lei Chen, Wei-Jun Zhu, Hui-Huang Huang, Jun-Liang Fu, Wei-Wei Chen, Ming Shi, Tong Zhang, Hao Wu, Fu-Sheng Wang
To date, there have been no reports characterizing the Human Immunodeficiency Virus-1 (HIV-1) in the semen of Chinese men having sex with men (MSM) with early infection of HIV-1.We examined genetic diversity and viral load of HIV-1 in the seminal compartments and blood of Chinese MSM with early HIV-1 infection in this study. Viral load and genetic diversity of HIV-1 in paired samples of semen and blood were analyzed in seven homosexual patients who were with early HIV-1 infection. Levels of HIV-1 RNA and DNA were tested by real-time PCR assays...
May 13, 2017: Microbiology and Immunology
Akio Abe, Ryutaro Nishimura, Asaomi Kuwae
Bordetella bronchiseptica infects a wide variety of mammals, and the type III secretion system (T3SS) is involved in the long-term colonization of Bordetella in the trachea and lung. The T3SS translocates virulence factors (commonly referred to as effectors) into host cells, leading to alteration of the host physiological function. The Bordetella effectors BopN and BteA are known to have roles in the up-regulation of IL-10 and the cytotoxicity, respectively. Nevertheless, the mechanism by which BopN is translocated into host cells has not been examined in sufficient detail...
May 13, 2017: Microbiology and Immunology
Souichi Nukuzuma, Chiyoko Nukuzuma, Masanori Kameoka, Shigeki Sugiura, Kazuo Nakamichi, Takafumi Tasaki, Tsutomu Takegami
JC polyomavirus (JCPyV) is the causative agent of the demyelinating disease in the central nervous system, known as PML in immunocompromised patients. Moreover, PML occurred in patients treated with natalizumab for MS or Crohn's disease as natalizumab-related PML. Many antiviral agents are being investigated due to the lack of drugs currently available for PML. The previous study demonstrated that the topoisomerase I inhibitors topotecan and β-lapachone have inhibitory effects on JCPyV replication in IMR-32 cells...
May 2, 2017: Microbiology and Immunology
Yan Shen, Xin Xie, Zhuolun Li, Yan Huang, Li Ma, Xinhe Shen, Yanyue Liu, Yuxia Zhao
IL-17 plays a key role in a variety of autoimmune diseases. MCP-1 is involved in the infiltration of mononuclear cells of myocardium in VMC. However, the relationship between IL-17 and MCP-1 in myocardial injury remains unclear. In this study, expression of MCP-1 mRNA and protein in cardiac myocytes was detected with qRT-PCR and ELISA, respectively. It was found that IL-17A induced MCP-1 expression in a dose- and time-dependent manner in cardiac myocytes, which could be blocked by IL-17A and IL-17RA neutralizing antibodies...
July 2017: Microbiology and Immunology
Yeon-Sook Kim, Yeon-Joo Choi, Kyung-Min Lee, Kyu-Joong Ahn, Heung-Chul Kim, Terry Klein, Ju Jiang, Allen Richards, Kyung-Hee Park, Won-Jong Jang
A Rickettsia sp. was isolated from the blood of a patient with an acute febrile illness using the shell vial technique; the isolate was named CN45Kr and was identified by molecular assay as Rickettsia monacensis, which was first recognized as a pathogen in Spain. Sequencing analysis showed that the gltA sequence of the isolate was identical to that of Rickettsia sp. IRS3. The ompA-5mp fragment sequence showed 100% identity to those of R. monacensis and Rickettsia sp. In56 and ompA-3pA In56 and 100% identity to that of Rickettsia sp...
July 2017: Microbiology and Immunology
Qiangde Duan, Weiping Zhang
Heat-stable toxin type I (STa)-ovalbumin chemical conjugates are currently used as the only coating antigen in ELISA to titrate anti-STa antibodies for ETEC vaccine candidates. STa-ovalbumin chemical conjugation requires STa toxin purification, a process that can be carried out by only a couple of laboratories and often with a low yield. Alternative ELISA coating antigens are needed for anti-STa antibody titration for ETEC vaccine development. In the present study, we genetically fused STa toxin gene (three copies) to a modified chicken ovalbumin gene for genetic fusion 3×STa-ovalbumin, and examined application of this fusion protein as an alternative coating antigen of anti-STa antibody titration ELISA...
July 2017: Microbiology and Immunology
Nanae Minami, Takayuki Abe, Lin Deng, Chieko Matsui, Takasuke Fukuhara, Yoshiharu Matsuura, Ikuo Shoji
Interferon-stimulated gene 15 (ISG15), a ubiquitin-like protein, is induced by type I INF. Although several groups have reported ISGylation of the HCV NS5A protein, it is still unclear whether ISGylation of NS5A has anti- or pro-viral effects in hepatitis C virus (HCV) infection. In the present study, the role of ISGylation-independent, unconjugated ISG15 in HCV infection was examined. Immunoprecipitation analyses revealed that ISG15 interacts specifically with NS5A domain I. ISG15 mutants lacking the C-terminal glycine residue that is essential for ISGylation still interacted with NS5A protein...
July 2017: Microbiology and Immunology
Narges Zeinalzadeh, Ali Hatef Salmanian, Goli Goujani, Jafar Amani, Ghasem Ahangari, Asal Akhavian, Mahyat Jafari
Enterotoxigenic Escherichia Coli (ETEC) strains are the commonest bacteria causing diarrhea in children in developing countries and travelers to these areas. Colonization factors (CFs) and enterotoxins are the main virulence determinants in ETEC pathogenesis. Heterogeneity of CFs is commonly considered the bottleneck to developing an effective vaccine. It is believed that broad spectrum protection against ETEC would be achieved by induced anti-CF and anti-enterotoxin immunity simultaneously. Here, a fusion antigen strategy was used to construct a quadrivalent recombinant protein called 3CL and composed of CfaB, a structural subunit of CFA/I, and CS6 structural subunits, LTB and STa toxoid of ETEC...
July 2017: Microbiology and Immunology
Hiroshi Oki, Takuya Yazawa, Yasuko Baba, Yumi Kanegae, Hanako Sato, Seiko Sakamoto, Takahisa Goto, Izumu Saito, Kiyoyasu Kurahashi
Pulmonary emphysema impairs quality of life and increases mortality. It has previously been shown that administration of adenovirus vector expressing murine keratinocyte growth factor (KGF) before elastase instillation prevents pulmonary emphysema in mice. We therefore hypothesized that therapeutic administration of KGF would restore damage to lungs caused by elastase instillation and thus improve pulmonary function in an animal model. KGF expressing adenovirus vector, which prevented bleomycin-induced pulmonary fibrosis in a previous study, was constructed...
July 2017: Microbiology and Immunology
Kosuke Kataoka, Yoshiko Fukuyama, David E Briles, Tatsuro Miyake, Kohtaro Fujihashi
To develop safe vaccines for inducing mucosal immunity to major pulmonary bacterial infections, appropriate vaccine antigens (Ags), delivery systems and nontoxic molecular adjuvants must be considered. Such vaccine constructs can induce Ag-specific immune responses that protect against mucosal infections. In particular, it has been shown that simply mixing the adjuvant with the bacterial Ag is a relatively easy means of constructing adjuvant-based mucosal vaccine preparations; the resulting vaccines can elicit protective immunity...
June 2017: Microbiology and Immunology
Neha Lohia, Manoj Baranwal
Influenza vaccine development is considered to be complicated and challenging. Constantly evolving influenza viruses require continuous global monitoring and reformulation of the vaccine strains. Peptide conserved among different strains and subtypes of influenza A virus are well-thought-out to be attractive targets for development of cross protective influenza vaccine based on cellular response. Three highly conserved (> 90%) matrix 1 peptides ILGFVFTLTVPSERGLQRRRF (PM 1), LIRHENRMVLASTTAKA (PM 2) and LQAYQKRMGVQMQR (PM 3) containing multiple T cell epitopes have been assessed for their immunogenic potential in vitro, subjecting peripheral blood mononuclear cells from healthy volunteers to repetitive stimulation of these chemically synthesised peptides and measuring their interferon (IFN)-γ level and proliferation by ELISA (Enzyme-linked immunosorbent assay) and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium bromide) assay, respectively...
April 21, 2017: Microbiology and Immunology
Yuka Sato, Tetsuo Koshizuka, Kei Ishibashi, Koichi Hashimoto, Ken Ishioka, Kazufumi Ikuta, Shin-Ichi Yokota, Nobuhiro Fujii, Tatsuo Suzutani
The suppressor of cytokine signaling (SOCS) family consists of 8 members and suppresses various cytokine signaling pathways including interferon (IFN) signaling. Therefore, some viruses have evolved molecular mechanisms to induce SOCS proteins in order to escape from host immunity. Herpes simplex virus type 1 (HSV-1) has a mechanism for escaping from type I IFN by induction of SOCS1 and SOCS3 as well. In this study, expression of 8 members of the SOCS family by HSV-1 infection was comparatively analyzed by quantitative reverse transcription PCR...
April 17, 2017: Microbiology and Immunology
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