ABSTRACT
Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin-Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.
Subject(s)
Antiviral Agents/pharmacology , Fatty Acids, Unsaturated/pharmacology , Gastropoda/chemistry , Mucus/chemistry , Orthomyxoviridae/drug effects , Virus Replication/drug effects , Animals , Antiviral Agents/metabolism , Dogs , Fatty Acids, Unsaturated/metabolism , Gastropoda/metabolism , Humans , Influenza, Human/virology , Madin Darby Canine Kidney Cells , Mucus/metabolism , Orthomyxoviridae/physiologyABSTRACT
Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin-Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.
ABSTRACT
Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin-Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.
ABSTRACT
Measles is a viral disease highly contagious spread by respiratory transmission. Although infection can be controlled by vaccination, numerous cases of measles have been registered in many areas of the world, highlighting the need for additional interventions. Terrestrial gastropods exude mucus on their body surface when traveling, to protect the body from mechanical injury, desiccation or contact with harmful substances. The mucus of mollusks has been studied as a source of new natural compounds with diverse biological activities. In this study, the antiviral activity of the mucus of the land slug P. boraceiensis was demonstrated in vitro using Vero cells infected with measles virus. The crude sample and four fractions were tested in cultures infected with measles virus and the antiviral activity was assessed by the cytopathic effect in infected cell cultures as well as by immunofluorescence and qPCR. Fractions 39 and 50 of the mucus from P. boraceiensis were analyzed by HPLC-DAD-ESI-MS/MS and infrared spectroscopy. A mixture of polyunsaturated fatty acids was found in the two fractions. A reduction in the growth of the measles virus was observed, measured by qPCR, with a protection index of 80% in Vero cells infected with measles and treated with fraction 39. Fraction 39 exhibited the best antiviral action in vitro and high contents of hydroxy-tritriacontapentaenoic acid and hydroxy-pentatriacontapentaenoic acid were found in this fraction.
Subject(s)
Antiviral Agents/pharmacology , Measles virus/drug effects , Mollusca/chemistry , Mucus/chemistry , Mucus/metabolism , Animals , Antiviral Agents/isolation & purification , Biological Products/chemistry , Biological Products/pharmacology , Carboxylic Acids/isolation & purification , Carboxylic Acids/pharmacology , Chlorocebus aethiops , Drug Discovery , Fatty Acids/isolation & purification , Fatty Acids/pharmacology , Tandem Mass Spectrometry , Vero Cells , Virus Replication/drug effectsABSTRACT
Measles is a viral disease highly contagious spread by respiratory transmission. Although infection can be controlled by vaccination, numerous cases of measles have been registered in many areas of the world, highlighting the need for additional interventions. Terrestrial gastropods exude mucus on their body surface when traveling, to protect the body from mechanical injury, desiccation or contact with harmful substances. The mucus of mollusks has been studied as a source of new natural compounds with diverse biological activities. In this study, the antiviral activity of the mucus of the land slug P. boraceiensis was demonstrated in vitro using Vero cells infected with measles virus. The crude sample and four fractions were tested in cultures infected with measles virus and the antiviral activity was assessed by the cytopathic effect in infected cell cultures as well as by immunofluorescence and qPCR. Fractions 39 and 50 of the mucus from P. boraceiensis were analyzed by HPLC-DAD-ESI-MS/MS and infrared spectroscopy. A mixture of polyunsaturated fatty acids was found in the two fractions. A reduction in the growth of the measles virus was observed, measured by qPCR, with a protection index of 80% in Vero cells infected with measles and treated with fraction 39. Fraction 39 exhibited the best antiviral action in vitro and high contents of hydroxy-tritriacontapentaenoic acid and hydroxy-pentatriacontapentaenoic acid were found in this fraction. (C) 2016 Elsevier B.V. All rights reserved.
Subject(s)
Virology , PathologyABSTRACT
OBJECTIVES: Phyllocaulis boraceiensis mucus has been studied as a potential source of new natural compounds that are capable of inducing proliferation and remodeling tissue. The aim of this study was to evaluate the clinical aspects of healing in the wounded mouse skin, which was treated with an ointment that was composed of mucus, which was released by P boraceiensis. MATERIALS & METHODS: Mice were submitted to a 1-cm dorsal excision. The control group (T1) was treated with papain; the T2 group was treated with papain that was associated with 0.18 µg/µL of mucus; and the T3 group was treated with papain that was associated with 0.012 µg/µL of mucus. RESULTS: Accelerated proliferation was observed after 3 days in the T3 group, presenting a high deposition of fibroblasts at the wound margin, whereas accelerated proliferation in the T1 group began 5 days after surgery. The T2 group presented inflammation during all periods of observation, and even when healing had already begun, the new tissue showed capillary fragility. Remodeling began after 4 days in the T3 group, whereas remodeling began after 6 days in the other groups. T3 showed edema, hyperemia, and bleeding only until the fifth day, and granulation and scar tissues intensely appeared from the 11th day forward. T1 and T2 groups exhibited edema, hyperemia, and bleeding until the 11th day, and granulation and scar tissues appeared after the 13th day. CONCLUSION: The healing process and wound closure were efficient after the daily application of 0.012 µg/µL P boraceiensis mucus.
Subject(s)
Biological Products/therapeutic use , Gastropoda , Mucus , Skin/blood supply , Wound Healing/physiology , Wounds and Injuries/therapy , Animals , Cell Proliferation/physiology , Disease Models, Animal , Female , Mice , Mice, Inbred BALB C , Ointments , Skin/injuries , Skin/physiopathology , Time FactorsABSTRACT
Gastrópodes terrestres exsudam muco através da superfície corporal quando se locomovem, para proteção do corpo contra injúria mecânica, dessecação ou contato com substâncias nocivas; a quantidade de muco aumenta quando são atacados por competidores, predadores ou alguns parasitas. Diversos autores têm estudado os componentes do muco do caracol gigante africano Achatina fulica, e demonstrado que o mesmo secreta substâncias com atividade bactericida que atuam tanto em bactérias Gram-positivas quanto em Gram-negativas, protegendo os gastrópodes que se locomovem nas mais diversas superfícies. Neste trabalho foi estudado o efeito microbiológico e bactericida do muco produzido pelas lesmas terrestres Phyllocaulis boraceiensis, mantidas em condições controladas de laboratório. Analisou-se o efeito bactericida e/ou bacteriostático de soluções teste frente às bactérias Gram−negativas (Pseudomonas aeruginosa e Escherichia coli) e Gram−positiva (Staphylococcus aureus), através de métodos de difusão em discos e de diluições em tubos. As propriedades do muco foram analisadas por métodos bioquímicos e os resultados serviram de base para as caracterizações das especificidades dessa(s)ação(ões) microbiológica(s).
Subject(s)
Biochemistry , Epithelial Cells , Mollusca/microbiologyABSTRACT
Gastrópodes terrestres exsudam muco através da superfície corporal quando se locomovem, para proteção do corpo contra injúria mecânica, dessecação ou contato com substâncias nocivas; a quantidade de muco aumenta quando são atacados por competidores, predadores ou alguns parasitas. Diversos autores têm estudado os componentes do muco do caracol gigante africano Achatina fulica, e demonstrado que o mesmo secreta substâncias com atividade bactericida que atuam tanto em bactérias Gram-positivas quanto em Gram-negativas, protegendo os gastrópodes que se locomovem nas mais diversas superfícies. Neste trabalho foi estudado o efeito microbiológico e bactericida do muco produzido pelas lesmas terrestres Phyllocaulis boraceiensis, mantidas em condições controladas de laboratório. Analisou-se o efeito bactericida e/ou bacteriostático de soluções teste frente às bactérias Gram−negativas (Pseudomonas aeruginosa e Escherichia coli) e Gram−positiva (Staphylococcus aureus), através de métodos de difusão em discos e de diluições em tubos. As propriedades do muco foram analisadas por métodos bioquímicos e os resultados serviram de base para as caracterizações das especificidades dessa(s)ação(ões) microbiológica(s).
Subject(s)
Biochemistry , Epithelial Cells , Mollusca/microbiologyABSTRACT
Um estudo comparativo sobre a ação de diversos fixadores utilizados em microscopia eletrônica de transmissão (MET) e microscopia de luz (ML) foi realizado, a fim de analisar a preservação de estruturas celulares ao microscópio eletrônico de transmissão (MET). Fragmentos de fígado de camundongo foram fixados em 5 diferentes fixadores: o fixador de Karnovsky, glutaraldeído e paraformaldeído utilizados no processamento para MET e o formaldeído comercial e líquido de Bouin utilizados no processamento para ML. Após a fixação, os fragmentos foram pós-fixados com tetróxido de ósmio e contrastastados com acetato de uranila. A seguir foram desidratados e incluídos em resina Epon. Os cortes ultrafinos mostraram que os fragmentos fixados com Karnovsky e glutaraldeído apresentaram melhor preservação das estruturas celulares e menor extração. O paraformaldeído produziu alguns artefatos de má fixação e extração pelo fato de formar menos ligações cruzadas que o glutaraldeído. Os fixadores formaldeído comercial e o líquido de Bouin utilizados em microscopia de luz mostraram que não são adequados ao uso em MET, pois são considerados fixadores coagulantes e produzem extensa extração dos componentes celulares. Comparando-se as imagens obtidas com os fixadores utilizados, o fixador de Karnovsky e o glutaraldeído mostraram melhor preservação e maior similaridade na morfologia
