<ORGINAL ARTICLE>Comparison of the characteristics of a hydrophilic variant of Streptococcus gordonii ATCC 10558 and a hydrophilic variant of Streptococcus mutans Ingbritt

Streptococcus mutans Ingbrittからhydrophihc variantを得るのと同様の方法にてStreptococcus gordonii ATCC 10558よりhydrophilic variantを得た。S. gordonn ATCC 10558のhydrophihc variantは親株に比較してhydrophobicityは低下していたが,唾液コートハイドロキシアパタイト(S-HA)に対する付着性は低下しなかった。S. gordonii ATCC 10558のhydrophilic variantは親株に比較して培地中に多くの種類のタンパク質を産生した。これらのことより,S gordonii ATCC 10558のS-HAに対する付着因子はS. mutans Ingbrittの付着因子と異なり菌体表層のhydrophobicityに関与する物質とは異なることが示唆された。We isolated the hydrophilic variant of Streptococcus gordonn ATCC 10558 by the isolation procedure for the hydrophilic variant of Streptococcus mutans Ingbntt. The hydrophobicity of the variant of 5 S gordonn ATCC 10558 was lower than that of the parent strain, but adherence to salivary coated hydroxyapatite (S-HA) was not lower. The hydrophilic variant of 5 gordonn ATCC 10558 released more quantity of proteins of various molecular weights into the culture supernatant than parent strain. The findings suggest that the adhesin of S. gordonii ATCC 10558 to S-HA differs from the components causing cell surface hydrophobicity in S mutans Ingbntt

The Diagnosis and Blistering Mechanisms of Mucous Membrane Pemphigoid

Mucous membrane pemphigoid (MMP) is a mucous membrane-dominated autoimmune subepithelial blistering disease that is caused by autoantibodies against various autoantigens in basement membrane zone (BMZ) proteins, including collagen XVII (COL17). Clinicians face diagnostic problems in detecting circulating antibodies and targeted antigens in MMP. The diagnostic difficulties are mainly attributed to the low titers of MMP autoantibodies in sera and to heterogeneous autoantigens. Additionally, no unanimous diagnostic criteria have been drawn for MMP, which can result in delayed diagnoses or misdiagnoses. This review aims to integrate and present currently available data to clarify diagnostic strategies and to present diagnostic criteria for MMP. The ultimate blistering mechanism in MMP has not been elucidated, and such mechanism is especially obscure in COL17-type MMP. In bullous pemphigoid (BP), which is the most common autoimmune subepidermal blistering disease, some patients show oral lesion as well as predominant skin lesions. However, there is no fundamental explanation for the onset of oral lesions in BP. This article summarizes innovative research perspectives on the pathogenesis of oral lesions in pemphigoid. Finally, we propose a potential pathogenesis for COL17-type MMP

Purification and characterization of hemolysin from Prevotella oris

AbstractWe observed hemolytic activity in culture supernatant of Prevotella oris. Results from growth-phase experiments show that hemolysin production increased during the logarithmic growth phase and decreased during the stationary phase. The hemolysin produced by P. oris was purified from the culture supernatant by ultrafiltration, diethylaminoethyl (DEAE) and carboxymethyl (CM) ion-exchange chromatography, and gel filtration chromatography; further, we investigated the purified hemolysin characteristics, including its ability to lyse human, horse, sheep, and rabbit erythrocytes. The purified hemolysin was observed as a single, 16-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. The specific activity was obtained by concentrating the purified hemolysin by 9200 fold. Although hemolysin was inactivated by heat treatment, ethylenediaminetetraacetic acid (EDTA), l-cysteine, dithiothreitol (DTT), and 2-mercaptoethanol enhanced its activity. Further, treatments using trypsin, MgCl2, CaCl2, and cholesterol did not affect its hemolytic activity. A pH of 6.0 was optimal for inducing the hemolysin activity. To the best of our knowledge, this is the first report describing the purification and characterization of hemolysin produced by P. oris

Development of Film Dosage Form Containing Allopurinol for Prevention and Treatment of Oral Mucositis

Film dosage forms (FDs) containing allopurinol (AP) were prepared using a casting method with water-soluble polysaccharides, such as sodium alginate (ALG), and the release profile of AP from FDs was investigated in limited dissolution medium. Some ALGs were able to form FDs incorporating AP, and the thickness was about 50 μm. All FDs were easy to handle, though the rheological properties varied with ALG species. AP was homogenously present throughout the FDs and was released with disintegration in 10 mL of physiological saline. These results confirmed that FDs are useful for preventing or treating localized problems in the oral cavity, such as mucositis. FDs are also useful for administering drugs to cancer patients receiving chemotherapy and/or radiotherapy

Prevotella oris溶血素の特徴とその赤血球膜との相互作用(The characteristics of Prevotella oris hemolysin and its interaction with the erythrocyte membrane)

P.orisはSDSポリアクリルアミドゲル電気泳動(SDS-PAGE)上で16kDaバンドとして観察される蛋白質溶血素を産生する。P.oris溶血素のN末端アミノ酸配列を分析した。7N末端アミノ酸を特定し、P.oris溶血素が短鎖ペプチド配列を有すると推定した。溶血素のアミノ酸配列は他の細菌性蛋白やペプチドと共通の配列ではなかった。P.oris溶血素は溶血前に温度依存性に赤血球と結合することが明らかになった。この溶血活性は赤血球をトリプシンまたはグリコシダーゼで処理した場合に阻害された。コレステロールは活性には影響しなかった。トリプシン処理した赤血球膜の特徴からSDS-PAGEにおいて46kDa糖蛋白が消失した。以上より、46kDa赤血球膜糖蛋白は溶血素の結合部位であることが示唆された。本研究はP.oris溶血素のN末端アミノ酸配列とその赤血球膜上の結合部位に関する初めての報告である。P.orisはSDSポリアクリルアミドゲル電気泳動(SDS-PAGE)上で16kDaバンドとして観察される蛋白質溶血素を産生する。P.oris溶血素のN末端アミノ酸配列を分析した。7N末端アミノ酸を特定し、P.oris溶血素が短鎖ペプチド配列を有すると推定した。溶血素のアミノ酸配列は他の細菌性蛋白やペプチドと共通の配列ではなかった。P.oris溶血素は溶血前に温度依存性に赤血球と結合することが明らかになった。この溶血活性は赤血球をトリプシンまたはグリコシダーゼで処理した場合に阻害された。コレステロールは活性には影響しなかった。トリプシン処理した赤血球膜の特徴からSDS-PAGEにおいて46kDa糖蛋白が消失した。以上より、46kDa赤血球膜糖蛋白は溶血素の結合部位であることが示唆された。本研究はP.oris溶血素のN末端アミノ酸配列とその赤血球膜上の結合部位に関する初めての報告である

<ORIGINAL>Quantification of Porphyromonas gingivalis by real time PCR : new primers targeting the rgpA and rgpB gene encoding RGP

We designed new primers for the quantification of Porphyromonas gingivalis by real time PCR. The new primer set targeted the rgpA and rgpB genes that encode arginine specific cysteine proteinase (Arggingipain or Rgp), one of the putative pathogenic factors of P. gingivalis. The PCR product obtained using our primers showed no by-products by melting curve analysis. The PCR product sequence showed no significant matches to other sequences by BLAST searching of genetic databases except for matches to P. gingivalis rgpA and rgpB sequence, and could not be amplified from template derived from other oral bacteria apart from P. gingivalis. Therefore, we concluded that our primers were specific for P. gingivalis rgpA and rgpB, and could be used to quantity from 10^3 to 10^7 P. gingivalis cells when applied to real time PCR

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Bacillus nattoがCandida albicansとC.stellatoideaに対してはその発育を阻止するが,他のCandida種の発育は阻止しない抗真菌物質を菌体外に産生することを確認した。B.nattoの培養上澄より活性炭の添加と限外ろ過(分子量 10,000)処理により粗抗カンジダ物質(ACS)を得た。粗ACSは供試したC.albicans154株の全株の発育を阻止したが,C.tropicalisの全株(5株)は発育が阻止されないことをペーパーディスク法により確認した。この方法により,粗ACSはこの2種のカンジダを明確に分別できることが判明した。粗ACSのC.albicansに対するMICは400μg/mlであったが,C.tropicalisは12,800μg/mlでも発育は阻止されなかった。-20℃に保存した粗ACSと4℃又は-20℃に保存した粗ACS溶液は初期の活性を8週間保持した。粗ACSのC.albicansに対する作用は主に静菌的であり,また弱く殺菌的にも作用した。この結果は粗ACSで処理したC.albicansの形態変化とも一致した。粗ACS含有サブロー寒天平板上のC.albicansの形態は,初めに一部の菌体が膨化し,次いで破壊される像が観察された。また,粗ACS含有液体培地中におけるC.albicansの形態は破壊され遊離した細胞質様物質が菌体周囲に存在し,多くの菌体はグラム陰性に染色された。以上の成績から,より簡単で迅速かつ正確なC.albicansの同定が,このペーパーディスク法と他の同定法とを併用することによって成し得ることが示唆された。Bacillus natto produced antifungal substance extracelluraly which inhibited the growth of Candida albicans and C. stellatoidea, but did not inhibit the growth of other Candida species. This antifungal substance was obtained from the culture filtrate by a procedure including the addition of active carbon and molecular filtration (MW 10,000), and this substance was designated as crude Anti-Candida substance (ACS). By paper disk method with crude ACS, the inhibitory zone was detected in all strains of C. albicans but not detected in any strains of C. tropicalis. A clear distinguishment between these two species was obtained by this method. MIC of crude ACS against C. albicans was 400 μg/ml but C. tropicalis growth was not inhibited at 12,800 μg/ml of crude ACS. Crude ACS which was stored at -20℃ and crude ACS solution which was stored at 4℃ or -20℃ retained its original inhibitory activity for 8 weeks. The action of crude ACS against C. albicans was mainly bacteriostatic and weakly bacteriocidal. This result was also identical with the morphological aspect of C. albicans treated with crude ACS. Regarding the morphological aspects of C. albicans on crude ACS-Sabouraud\u27s dextrose agar plate some organisms were swollen at first, and as a secondary step these were destroyed. In conclusion, this study suggests that a more easy, rapid, and exact identification of C. albicans can be done by the utilization this paper disk method in parallel with other identification methods

The Impact of PNPLA3 rs738409 Genetic Polymorphism and Weight Gain ≥10 kg after Age 20 on Non-Alcoholic Fatty Liver Disease in Non-Obese Japanese Individuals.

Non-alcoholic fatty liver disease (NAFLD) in non-obese individuals is inadequately elucidated. We aim to investigate the impact of known genetic polymorphisms on NAFLD and the interaction between genetic risks and weight gain on NAFLD in obese and non-obese Japanese individuals. A total of 1164 participants who received health checkups were included. Participants with excessive alcohol consumption, with viral hepatitis or other inappropriate cases were excluded. Fatty liver was diagnosed by ultrasonography. Participants with a body mass index (BMI) of <18.5 kg/m2, 18.5-22.9 kg/m2, 23.0-24.9 kg/m2 and ≥25 kg/m2 were classified underweight, normal weight, overweight and obese, respectively. Self-administered questionnaire for lifestyle was assessed and a total of 8 previously reported genetic polymorphisms were chosen and examined. In all, 824 subjects were enrolled. The overall prevalence of NAFLD was 33.0%: 0% in underweight, 15.3% in normal weight, 41.1% in overweight and 71.7% in obese individuals. The prevalence of NAFLD is more affected by the G allele of patatin-like phospholipase domain-containing protein 3 (PNPLA3) rs738409 in normal weight (odds ratio (OR) 3.52; 95%-CI: 1.42-8.71; P = 0.0063) and in overweight individuals (OR 2.60; 95%-CI: 1.14-5.91; P = 0.0225) than in obese individuals (not significant). Moreover, the G allele of PNPLA3 rs738409 and weight gain ≥10 kg after age 20 had a joint effect on the risk of NAFLD in the normal weight (OR 12.00; 95% CI: 3.71-38.79; P = 3.3×10-5) and the overweight individuals (OR 13.40; 95% CI: 2.92-61.36; P = 0.0008). The G allele of PNPLA3 rs738409 is a prominent risk factor for NAFLD and the interaction between the PNPLA3 rs738409 and weight gain ≥10 kg after age 20 plays a crucial role in the pathogenesis of NAFLD, especially in non-obese Japanese individuals

口腔内偏性嫌気性糖非分解性細菌Eubacteriumその意義と展望

Historically, it has been suggested for a long time that there are so many uncultured, undescribed and unknown bacterial species including VNC (viable but non-cultivable) in the human oral cavity, especially infectious lesions such as periodontal pockets. All these strains are asaccharolytic anaerobic gram-positive rods, and have been proven to be difficult to culture and unreactive in the conventional biochemical tests. The molecular systematic techniques including 16S rRNA gene sequencing analysis and DNA-DNA hybridization method have demonstrated that they are mainly Eubacterium and/or the related bacteria with Eubacterium, phylogenetically. And recently, many novel bacterial genera and species, such as genus Slackia, Eggerthella, Cryptobacterium and Mogibacterium, have been proposed on the basis of phylogenetic data for these bacterial strains. A complete description of the microbial flora associated with oral infections lead to the potential benefits for the elucidation of disease causation and also the development of novel diagnostic tools. In this paper, we summarize the recent knowledge of these bacteria species, and also discuss the etiologic roles of these bacteria in human oral infections

Porphyromonas gingivalisの自己融解酵素変異株の性状

The results of a BLAST search for autolysin indicated that one ORF was indentified as an autolysin which had 39% homology (58/147 amino acids) with the N-acetylmuramoyl-L-alanine amidase like protein of Clostridium tetani. This autolysin mutant of Porphyromonas gingivalis was constructed by alleic exchange. The growth curve of the autolysin mutant was the same as that of parent strain. However, the morphology of the autolysin mutant altered into chain forms at the early logphase. These results indicate that this autolysin may play an important role in cell division in P. gingivalis