Corrigendum: Recombinant Porphyromonas gingivalis FimA preproprotein expressed in Escherichia coli is lipidated and the mature or processed recombinant FimA protein forms a short filament in vitro

2011 ◽  
Vol 57 (1) ◽  
pp. 68
Author(s):  
Mikio Shoji ◽  
Atsutoshi Yoshimura ◽  
Hidenobu Yoshioka ◽  
Akemi Takade ◽  
Yasuko Takuma ◽  
...  

2016 ◽  
Vol 17 (4) ◽  
pp. 282-285 ◽  
Author(s):  
Deema J Farsi ◽  
Anne Tanner

ABSTRACT Aim To determine the sensitivity of Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia to triclosan, and determine if these bacteria develop resistance to triclosan upon prolonged exposure. Materials and methods Susceptibility to triclosan was tested against three periodontal pathogens P. gingivalis, P. intermedia, and T. forsythia. Escherichia coli strains sensitive and resistant to triclosan were used as biological controls to confirm the efficacy of triclosan in the assays. Agar plates were prepared locally with vitamin K and hemin-supplemented medium. Results Porphyromonas gingivalis and P. intermedia did not grow on plates containing ≥2 μg/ml triclosan, while T. forsythia did not grow on ≥1.66 μg/ml. Colonies of P. intermedia resistant to triclosan developed after prolonged incubation at 2 μg/ml, but this resistance disappeared during subculture in the absence of triclosan. Conclusion No significant resistance to triclosan was detected for these species. Clinical significance Dental products containing triclosan can be beneficial in controlling periodontal disease. How to cite this article Farsi D, Tanner A. In vitro Resistance Testing of Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia to Triclosan. J Contemp Dent Pract 2016;17(4):282-285.



2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Tatiane Oliveira ◽  
Camila A. Figueiredo ◽  
Carlos Brito ◽  
Alexander Stavroullakis ◽  
Anuradha Prakki ◽  
...  

Allium cepaL. is known to possess numerous pharmacological properties. Our aim was to examine thein vitroeffects ofAllium cepaL. extract (AcE) onPorphyromonas gingivalisLPS andEscherichia coliLPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated byPgLPS (1 μg/mL) andE. coliLPS (1 μg/mL) in the presence or absence of different concentrations of AcE (10–1000 μg/mL) for 5 days at 37°C/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000 μg/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest thatAllium cepaL. extract could be used forin vitrostudies onPorphyromonas gingivalisLPS andEscherichia coliLPS-stimulated osteoclast precursor cells.



2021 ◽  
Author(s):  
Auygna Pamyda Gomes da Silva ◽  
Abigail Eduarda de Miranda Magalhães ◽  
Maria Amélia Paiva Ferruccio ◽  
Tayonara dos Santos Melo ◽  
Tuanne dos Santos Melo

Introdução: A Plantago major L. é popularmente conhecida como tanchagem pertence à família Plantaginacea. Tem estrutura perene com folhas rosetadas, as flores e folhas jovens são comestíveis. Utilizada na medicina popular para o tratamento de lesões na pele, gengivite, abcessos, dor de dente. Possui grande variedade de bioativos como alcalóides, terpenos, flavonóides, vitaminas. A sua atividade funcional é relacionada a presença desses compostos químicos.(1) Objetivos: Realizar um levantamento bibliográfico para identificar a atividade antimicrobiana de P. major. Métodos: Trata-se de uma revisão bibliográfica do tipo integrativa realizada por busca nas bases de dados Pubmed, SciELO e BVS, trabalhos indexados no período de 2016 a 2021. Foram utilizados para busca dos artigos os seguintes descritores “Plantago major” e “antimicrobial”. Como critério de inclusão foram pesquisados artigos em inglês, espanhol e português, disponíveis na íntegra, estudos com informações referentes ao uso da espécie como agente antimicrobiano. Os critérios de exclusão foram os artigos duplicados, revisões bibliográficas. Resultados: Foram encontrados 18 artigos, após realizar uma análise criteriosa do material, foram incluídos na revisão 6 trabalhos. Um dos trabalhos sugeriu a utilização de extrato de P. major como enxaguante bucal para o tratamento de mucosite oral.(2) Em outro estudo a aplicação do extrato de P. major mostrou atividade antibacteriana contra Pseudomonas tolaasii, resultando um maior diâmetro inibitório. Também apresentou inibição de crescimento contra Pseudomonas aeruginosa utilizando a técnica de disco, com o aparecimento de halos de inibição de crescimento maiores que 1 cm. No ensaio de citotoxicidade, o grau de hemólise foi classificado como baixo (5%), logo, indicando a segurança no uso da tanchagem(3) Utilizando sementes de P. major aplicadas em nanoparticulas apresentou atividade aceitável contra Micrococcus luteus, Escherichia coli e Penicillium digitatum. Além desses patógenos, outro estudo evidenciou que o extrato etanólico (75%;100%) de tanchagem apresentou efeito antibacteriano in vitro contra Porphyromonas gingivalis, apresentando halos de inibição.(4) Também foi obtido resultados contra Cândida albicans, aplicando extrato de P. major, com aucubina e baicaleína, atuando na hidrofobicidade de C. albicans, demonstrando eficácia como antifúngico, sugerindo ser um potencial promissor para infecções relacionadas ao biofilme por C. Albicans.(5) Conclusões: A partir desses resultados é possível identificar a possibilidade do uso de Plantago major para fins terapêuticos, fitoterápicos ou nutricionais sendo necessário realizar mais estudos quanto a sua atividade antimicrobiana além de testes quanto à sua toxicidade.



2010 ◽  
Vol 56 (11) ◽  
pp. 959-967 ◽  
Author(s):  
Mikio Shoji ◽  
Atsutoshi Yoshimura ◽  
Hidenobu Yoshioka ◽  
Akemi Takade ◽  
Yasuko Takuma ◽  
...  

The gram-negative anaerobic bacterium Porphyromonas gingivalis is an etiologically important pathogen for chronic periodontal diseases in adults. Our previous study suggested that the major structural components of both Fim and Mfa fimbriae in this organism are secreted through their lipidated precursors. In this study, we constructed Escherichia coli strains expressing various fimA genes with or without the 5′-terminal DNA region encoding the signal peptide, and we determined whether lipidation of recombinant FimA proteins occurred in E. coli. Lipidation occurred for a recombinant protein from the fimA gene with the 5′-terminal DNA region encoding the signal peptide but not for a recombinant protein from the fimA gene without the signal-peptide-encoding region, as revealed by [3H]palmitic acid labeling experiments. A TLR2-dependent signaling response was induced by the recombinant protein from the fimA gene with the signal-peptide-encoding region but not by a recombinant protein from the fimA gene with the signal-peptide-encoding region that had a base substitution causing an amino acid substitution (C19A). Electron microscopic analysis revealed that recombinant FimA (A-47 – W-383) protein was autopolymerized to form filamentous structures of about 80 nm in length in vitro. The results suggest that FimA protein, a major subunit of Fim fimbriae, is transported to the outer membrane by the lipoprotein sorting system, and a mature or processed FimA protein on the outer membrane is autopolymerized to form Fim fimbriae.





Author(s):  
О.В. Шамова ◽  
М.С. Жаркова ◽  
П.М. Копейкин ◽  
Д.С. Орлов ◽  
Е.А. Корнева

Антимикробные пептиды (АМП) системы врожденного иммунитета - соединения, играющие важную роль в патогенезе инфекционных заболеваний, так как обладают свойством инактивировать широкий спектр патогенных бактерий, обеспечивая противомикробную защиту живых организмов. В настоящее время АМП рассматриваются как потенциальные соединения-корректоры инфекционной патологии, вызываемой антибиотикорезистентными бактериями (АБР). Цель данной работы состояла в изученим механизмов антибактериального действия трех пептидов, принадлежащих к семейству бактенецинов - ChBac3.4, ChBac5 и mini-ChBac7.5Nb. Эти химически синтезированные пептиды являются аналогами природных пролин-богатых АМП, обнаруженных в лейкоцитах домашней козы Capra hircus и проявляющих высокую антимикробную активность, в том числе и в отношении грамотрицательных АБР. Методы. Минимальные ингибирующие и минимальные бактерицидные концентрации пептидов (МИК и МБК) определяли методом серийных разведений в жидкой питательной среде с последующим высевом на плотную питательную среду. Эффекты пептидов на проницаемость цитоплазматической мембраны бактерий для хромогенного маркера исследовали с использованием генетически модифицированного штамма Escherichia coli ML35p. Действие бактенецинов на метаболическую активность бактерий изучали с применением маркера резазурина. Результаты. Показано, что все исследованные пептиды проявляют высокую антимикробную активность в отношении Escherichia coli ML35p и антибиотикоустойчивых штаммов Escherichia coli ESBL и Acinetobacter baumannii in vitro, но их действие на бактериальные клетки разное. Использован комплекс методик, позволяющих наблюдать в режиме реального времени динамику действия бактенецинов в различных концентрациях (включая их МИК и МБК) на барьерную функцию цитоплазматической мембраны и на интенсивность метаболизма бактериальных клеток, что дало возможность выявить различия в характере воздействия бактенецинов, отличающихся по структуре молекулы, на исследуемые микроорганизмы. Установлено, что действие каждого из трех исследованных бактенецинов в бактерицидных концентрациях отличается по эффективности нарушения целостности бактериальных мембран и в скорости подавления метаболизма клеток. Заключение. Полученная информация дополнит существующие фундаментальные представления о механизмах действия пролин-богатых пептидов врожденного иммунитета, а также послужит основой для биотехнологических исследований, направленных на разработку на базе этих соединений новых антибиотических препаратов для коррекции инфекционных заболеваний, вызываемых АБР и являющимися причинами тяжелых внутрибольничных инфекций. Antimicrobial peptides (AMPs) of the innate immunity are compounds that play an important role in pathogenesis of infectious diseases due to their ability to inactivate a broad array of pathogenic bacteria, thereby providing anti-microbial host defense. AMPs are currently considered promising compounds for treatment of infectious diseases caused by antibiotic-resistant bacteria. The aim of this study was to investigate molecular mechanisms of the antibacterial action of three peptides from the bactenecin family, ChBac3.4, ChBac5, and mini-ChBac7.5Nb. These chemically synthesized peptides are analogues of natural proline-rich AMPs previously discovered by the authors of the present study in leukocytes of the domestic goat, Capra hircus. These peptides exhibit a high antimicrobial activity, in particular, against antibiotic-resistant gram-negative bacteria. Methods. Minimum inhibitory and minimum bactericidal concentrations of the peptides (MIC and MBC) were determined using the broth microdilution assay followed by subculturing on agar plates. Effects of the AMPs on bacterial cytoplasmic membrane permeability for a chromogenic marker were explored using a genetically modified strain, Escherichia coli ML35p. The effect of bactenecins on bacterial metabolic activity was studied using a resazurin marker. Results. All the studied peptides showed a high in vitro antimicrobial activity against Escherichia coli ML35p and antibiotic-resistant strains, Escherichia coli ESBL and Acinetobacter baumannii, but differed in features of their action on bacterial cells. The used combination of techniques allowed the real-time monitoring of effects of bactenecin at different concentrations (including their MIC and MBC) on the cell membrane barrier function and metabolic activity of bacteria. The differences in effects of these three structurally different bactenecins on the studied microorganisms implied that these peptides at bactericidal concentrations differed in their capability for disintegrating bacterial cell membranes and rate of inhibiting bacterial metabolism. Conclusion. The obtained information will supplement the existing basic concepts on mechanisms involved in effects of proline-rich peptides of the innate immunity. This information will also stimulate biotechnological research aimed at development of new antibiotics for treatment of infectious diseases, such as severe in-hospital infections, caused by antibiotic-resistant strains.



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