Different hypersensitivities against homologous proteins of MGL_1304 in patients with atopic dermatitis

Publication date: Available online 24 June 2017
Source:Allergology International
Author(s): Takuma Kohsaka, Takaaki Hiragun, Kaori Ishii, Makiko Hiragun, Akiko Kamegashira, Michihiro Hide
BackgroundAtopic dermatitis (AD) is exacerbated by sweating, and the skin of most patients with AD are resided by Malassezia (M.) fungi. Recently, MGL_1304 produced by Malasseziaglobosa was identified as the major histamine releasing antigen in human sweat.MethodsThe full length cDNA of the counterpart of MGL_1304 in Malasseziarestricta (Mala r 8), was cloned by degenerate PCR and rapid identification of cDNA ends (RACE). Recombinant MGL_1304, and its counterparts, Mala s 8 (produced by Malasseziasympodialis) and Mala r 8 were prepared, and compared in their allergenicities by dot blot analysis and histamine release tests with sera and basophils of patients with AD.ResultsThe identities between MGL_1304 and Mala s 8, MGL_1304 and Mala r 8, and Mala s 8 and Mala r 8 were 68%, 78%, and 76%, respectively, in protein sequences. Dot blot analysis revealed that the level of IgE binding to Mala s 8 was higher than that of MGL_1304. However, histamine release tests revealed that MGL_1304 and Mala r 8 possessed higher activity than Mala s 8. In addition, the crude lysate of M. globosa showed higher histamine release ability than that of M. sympodialis.ConclusionsPatients with AD showed hypersensitivities against MGL_1304 and its homologs. However, the allergenicities of the homologs are variable and the histamine release activities may be different from the solid-phase binding activities for IgE. Sweat allergy should be carefully evaluated with biological activities of MGL_1304 and its homologs of other Malassezia fungi residing on the skin.



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Generation and characterization of a human oral squamous carcinoma cell line SCC-9 with CRISPR/Cas9-mediated deletion of the p75 neurotrophin receptor

Publication date: October 2017
Source:Archives of Oral Biology, Volume 82
Author(s): Ping Huang, Dongdong Tong, Jing Sun, Qing Li, Fenghe Zhang
ObjectiveTo investigate the importance of the p75 neurotrophin receptor (p75^NTR) in human tongue squamous carcinoma cells, we exploited the CRISPR/Cas9 technology to establish a p75^NTR-knockout SCC-9 cell line and to explore the effect on biological functions.Materials and methodsThe clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease (Cas9) system was used to generate genomic deletion mutants of p75^NTR in the tongue squamous carcinoma cell lines SCC-9. Single-guide RNA (sgRNA) sequences were designed to target the p75^NTR genomic sequence and were cloned into plasmid pGK1.1. The linearized vector was electroporated into SCC-9 cells and p75^NTR deletion was confirmed using Cruiser™ enzyme digestion and PCR amplification. SCC-9 clones with successful deletion of p75^NTR were identified and verified by sequencing and selected for functional testing in cell proliferation, invasion, migration, and colony-forming assays.ResultsCompared with control cells, p75^NTR-knockout SCC-9 cells showed significantly diminished abilities to proliferate, invade, migrate, and form colonies, indicating a reduction in pro-tumorigenic behavior.ConclusionThese data demonstrate, first, that the CRISPR/Cas9 system is a simplified method for generating p75^NTR knockouts with relatively high efficiency, and second, that deletion of p75^NTR suppresses several tumor-promoting properties of SCC-9 cells, suggesting that p75^NTR is a potential target for the development of novel therapies for tongue cancer.



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Fibulins and matrilins are novel structural components of the periodontium in the mouse

Publication date: October 2017
Source:Archives of Oral Biology, Volume 82
Author(s): Andrea Schubert, Boris Schminke, Nicolai Miosge
Periodontitis refers to inflammatory disease of the periodontal structures (the gingiva, dental cementum, periodontal ligament (PDL) and alveolar bone) that ultimately leads to their destruction. Whereas collagens are well-examined main components of the periodontium, little is known about the other structural proteins that make up this tissue. The aim of this study was to identify new extracellular matrix (ECM) components, including fibulins and matrilins, in the periodontium of mice.After sacrificing 14 mice (Sv/129 strain), jaws were prepared. Each tissue sample contained a molar and its surrounding alveolar bone. Immunohistochemistry was carried out on paraffin-embedded sections.Our results show that mice exhibit fibulin-3, -4 and -5 and matrilin-1, -2, -3 and -4 in PDL and in blood vessels of alveolar bone and PDL as well as in the pericellular matrix of osteocytes and cementocytes. In dental cementum, only fibulin-4 is expressed.For the first time, we show that fibulin-3, -4 and -5 and matrilin-1, -2, -3 and -4 are essential components of the periodontal tissues. Our findings indicate an association of these proteins with collagens and oxytalan fibers that might be of future interest in regenerative periodontitis therapy.



http://ift.tt/2sPt3ul

Generation and characterization of a human oral squamous carcinoma cell line SCC-9 with CRISPR/Cas9-mediated deletion of the p75 neurotrophin receptor

Publication date: October 2017
Source:Archives of Oral Biology, Volume 82
Author(s): Ping Huang, Dongdong Tong, Jing Sun, Qing Li, Fenghe Zhang
ObjectiveTo investigate the importance of the p75 neurotrophin receptor (p75^NTR) in human tongue squamous carcinoma cells, we exploited the CRISPR/Cas9 technology to establish a p75^NTR-knockout SCC-9 cell line and to explore the effect on biological functions.Materials and methodsThe clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease (Cas9) system was used to generate genomic deletion mutants of p75^NTR in the tongue squamous carcinoma cell lines SCC-9. Single-guide RNA (sgRNA) sequences were designed to target the p75^NTR genomic sequence and were cloned into plasmid pGK1.1. The linearized vector was electroporated into SCC-9 cells and p75^NTR deletion was confirmed using Cruiser™ enzyme digestion and PCR amplification. SCC-9 clones with successful deletion of p75^NTR were identified and verified by sequencing and selected for functional testing in cell proliferation, invasion, migration, and colony-forming assays.ResultsCompared with control cells, p75^NTR-knockout SCC-9 cells showed significantly diminished abilities to proliferate, invade, migrate, and form colonies, indicating a reduction in pro-tumorigenic behavior.ConclusionThese data demonstrate, first, that the CRISPR/Cas9 system is a simplified method for generating p75^NTR knockouts with relatively high efficiency, and second, that deletion of p75^NTR suppresses several tumor-promoting properties of SCC-9 cells, suggesting that p75^NTR is a potential target for the development of novel therapies for tongue cancer.



http://ift.tt/2s52qnX

Fibulins and matrilins are novel structural components of the periodontium in the mouse

Publication date: October 2017
Source:Archives of Oral Biology, Volume 82
Author(s): Andrea Schubert, Boris Schminke, Nicolai Miosge
Periodontitis refers to inflammatory disease of the periodontal structures (the gingiva, dental cementum, periodontal ligament (PDL) and alveolar bone) that ultimately leads to their destruction. Whereas collagens are well-examined main components of the periodontium, little is known about the other structural proteins that make up this tissue. The aim of this study was to identify new extracellular matrix (ECM) components, including fibulins and matrilins, in the periodontium of mice.After sacrificing 14 mice (Sv/129 strain), jaws were prepared. Each tissue sample contained a molar and its surrounding alveolar bone. Immunohistochemistry was carried out on paraffin-embedded sections.Our results show that mice exhibit fibulin-3, -4 and -5 and matrilin-1, -2, -3 and -4 in PDL and in blood vessels of alveolar bone and PDL as well as in the pericellular matrix of osteocytes and cementocytes. In dental cementum, only fibulin-4 is expressed.For the first time, we show that fibulin-3, -4 and -5 and matrilin-1, -2, -3 and -4 are essential components of the periodontal tissues. Our findings indicate an association of these proteins with collagens and oxytalan fibers that might be of future interest in regenerative periodontitis therapy.



http://ift.tt/2sPt3ul

Predictive value of body mass index to metabolic syndrome risk factors in Syrian adolescents

Obesity has become a serious epidemic health problem in both developing and developed countries. There is much evidence that obesity among adolescents contributed significantly to the development of type 2 dia...

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Intraoperative sialendoscopy to assist with and confirm repair of Stensen’s duct

Publication date: Available online 24 June 2017
Source:British Journal of Oral and Maxillofacial Surgery
Author(s): C.-B. Man, R. Patel, K. Karavidas




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