RESUMO
Piper is the largest genus of the Piperaceae family. The species of this genus have diverse biological activities and are used in pharmacopeia throughout the world. They are also used in folk medicine for treatment of many diseases in several countries including Brazil, China, India, Jamaica, and Mexico. In Brazil, Piper species are distributed throughout the national territory, making this genus a good candidate for biological activity screening. During our studies with Piper essential oils, we evaluated its activity against Rhizopus oryzae, the main agent of mucormycosis. The main compounds of seven Piper essential oils analyzed were Piper callosum-safrole (53.8%), P. aduncum-dillapiole (76.0%), P. hispidinervum-safrole (91.4%), P. marginatum-propiopiperone (13.2%), P. hispidum-γ-terpinene (30.9%), P. tuberculatum-(E)-caryophyllene (30.1%), and Piper sp.-linalool (14.6%). The minimum inhibitory concentration of Piper essential oils against R. oryzae ranged from 78.12 to >1250 µg/mL. The best result of total inhibition of biofilm formation was obtained with Piper sp. starting from 4.88 µg/mL. Considering the bioactive potential of EOs against planktonic cells and biofilm formation of R. oryzae could be of great interest for development of antimicrobials for therapeutic use in treatment of fungal infection.
RESUMO
Caprine arthritis-encephalitis virus (CAEV) belongs to Retroviridae family, Lentivirus genus. This virus infects caprine all over the world causing arthritis, encephalitis, mammitis and progressive emaciating. This research showed chimera's building made by mixing up CAEV p28, with glutaraldehyde, and CPSMV, purified through the chromotography in biogel and sephadex (G-150). After that, some measures in a spectrophometric were developed to absorbance at 280nm. Peaks, which contained chimera, were collected and submitted to SDS-PAGE, evidencing the band relative to itself. Groups of swiss mice were immunized with chimeric virus, purified CPSMV and with p28 protein using incomplete Freund Adjuvant. CPSMV and p28 specific antibodies recognized chimeric protein in Western Blotting and ELISA showing the efficacy of the method. The results showed the covalent coupling between CPSMV and CAEV p28 was successfully archieved, originating a stable molecule, which no disestablished the capside from CPSMV. Besides, it showed that chimeric virus presents more immunogenicity than protein p28 isolated. It's suggesting CPSMV can be used as a carrier molecule in the production of vaccines to the virus, which infect animals.
O vírus da Artrite-encefalite caprina (CAEV) pertence à família Retroviridae, gênero Lentivirus. O CAEV infecta caprinos do mundo inteiro causando artrite, encefalite, mamite, pneumonia e emagrecimento progressivo. Este trabalho mostra a formação de uma quimera construída através da mistura da p28 do CAEV com glutaraldeído e CPSMV, purificada por meio de cromatografia em biogel e sephadex G-150. As cromatografias foram monitoradas através de leituras em espectrofotômetro no comprimento de onda de 280nm, dos líquidos coletados nos tubos. Os picos contendo a quimera foram coletados e submetidos à eletroforese (SDS-PAGE), sendo assim evidenciada a banda correspondente à mesma. Grupos de camundongos swiss foram imunizados com o vírus quimérico (CPSMV + p28), com o vírus CPSMV purificado e com a proteína p28 do CAEV, utilizando o adjuvante de Freund incompleto. Os anticorpos específicos produzidos contra o CPSMV e p28 reconheceram a proteína quimérica em Western Blotting e em teste de ELISA. Os anticorpos contra o vírus quimérico apresentaram títulos mais elevados do que os anticorpos produzidos contra a p28, demonstrando que o vírus quimérico apresenta maior imunogenicidade do que a proteína p28 sozinha. Os resultados mostraram que o acoplamento covalente entre o CPSMV e a p28 do CAEV foi obtido com sucesso, originando uma molécula estável não comprometendo a estrutura do capsídeo do CPSMV. Desta forma, sugere-se que o CPSMV possa ser utilizado como molécula carreadora na produção de vacinas para vírus que infectam animais.
RESUMO
Caprine arthritis-encephalitis virus (CAEV) belongs to Retroviridae family, Lentivirus genus. This virus infects caprine all over the world causing arthritis, encephalitis, mammitis and progressive emaciating. This research showed chimera's building made by mixing up CAEV p28, with glutaraldehyde, and CPSMV, purified through the chromotography in biogel and sephadex (G-150). After that, some measures in a spectrophometric were developed to absorbance at 280nm. Peaks, which contained chimera, were collected and submitted to SDS-PAGE, evidencing the band relative to itself. Groups of swiss mice were immunized with chimeric virus, purified CPSMV and with p28 protein using incomplete Freund Adjuvant. CPSMV and p28 specific antibodies recognized chimeric protein in Western Blotting and ELISA showing the efficacy of the method. The results showed the covalent coupling between CPSMV and CAEV p28 was successfully archieved, originating a stable molecule, which no disestablished the capside from CPSMV. Besides, it showed that chimeric virus presents more immunogenicity than protein p28 isolated. It's suggesting CPSMV can be used as a carrier molecule in the production of vaccines to the virus, which infect animals.
O vírus da Artrite-encefalite caprina (CAEV) pertence à família Retroviridae, gênero Lentivirus. O CAEV infecta caprinos do mundo inteiro causando artrite, encefalite, mamite, pneumonia e emagrecimento progressivo. Este trabalho mostra a formação de uma quimera construída através da mistura da p28 do CAEV com glutaraldeído e CPSMV, purificada por meio de cromatografia em biogel e sephadex G-150. As cromatografias foram monitoradas através de leituras em espectrofotômetro no comprimento de onda de 280nm, dos líquidos coletados nos tubos. Os picos contendo a quimera foram coletados e submetidos à eletroforese (SDS-PAGE), sendo assim evidenciada a banda correspondente à mesma. Grupos de camundongos swiss foram imunizados com o vírus quimérico (CPSMV + p28), com o vírus CPSMV purificado e com a proteína p28 do CAEV, utilizando o adjuvante de Freund incompleto. Os anticorpos específicos produzidos contra o CPSMV e p28 reconheceram a proteína quimérica em Western Blotting e em teste de ELISA. Os anticorpos contra o vírus quimérico apresentaram títulos mais elevados do que os anticorpos produzidos contra a p28, demonstrando que o vírus quimérico apresenta maior imunogenicidade do que a proteína p28 sozinha. Os resultados mostraram que o acoplamento covalente entre o CPSMV e a p28 do CAEV foi obtido com sucesso, originando uma molécula estável não comprometendo a estrutura do capsídeo do CPSMV. Desta forma, sugere-se que o CPSMV possa ser utilizado como molécula carreadora na produção de vacinas para vírus que infectam animais.