No description available.

"Vulvovaginal candidiasis (VVC) is a mucosal infection caused by Candida species and represents one of the most common clinical problems in women of reproductive age (68,71). Annually in the United States there are approximately 13 million cases of VVC, resulting in 10 million gynecologic office visits per year (38). It is estimated that 75% of women will experience at least one episode in their lifetime, with a projected 50% of all women experiencing multiple episodes (23). Candida albicans is a dimorphic commensal organism of the urogenital and gastrointestinal tracts and has been identified as the main pathogenic agent in VVC, accounting for approximately 85-90% of patients with positive cultures (52). Despite extensive research, the invasive mechanism of vaginal yeast infections is not well understood. Traditionally it has been assumed that changes in the host vaginal environment promote the dimorphic transition from blastospore to hyphae, resulting in a shift from asymptomatic colonization to symptomatic vaginitis (28). In contrast to the normal, systemic immune response, which confers an aseptic environment for tissue and organs, immune responses at the mucosal level are designed to prevent tissue invasion and local disease while maintaining an indigenous flora that could be both beneficial and pathogenic (28). Since fungi are eukaryotic, the vital cellular mechanisms that are usually targeted by modern pharmacologic agents, such as DNA replication and protein translation, are either conserved or have a strong homology to their human orthologs. Obtaining a better understanding of natural fungal suppression mechanisms and molecules at the mucosal level may pave the way for the development of more efficacious drugs or preventative regiments. The mechanism by which the human immune system is able to resist fungal invasion at the vaginal mucosa is unknown. Our research was aimed at finding any host factors that might play a role in the suppression of or prevention of a fungal infection at the vaginal mucosa. In order to screen candidate molecules that might be important in this type of vaginal defense, we chose a pathogenic C. albicans strain, SC5314, to test fungal cell viability upon introduction of the candidate molecules. We have identified a host factor that exhibits strong fungicidal activity when organically extracted from both human and bovine serum albumins. Characterization of this factor through organic extractions and acetone separations reveal that this molecule is a non-polar lipid. Serum samples that have been thoroughly stripped of fatty acids and other lipophilic molecules show no apparent fungicidal activity in cell viability assays. Since the factor is extractable from both human and bovine serum albumins, it may be conserved among mammals. Identification and characterization of this molecule may play a pivotal role in understanding host-Candida interactions at the mucosal membrane interface. Due to its human origin, the use of this factor as an antifungal would be extremely advantageous in regards to FDA (Food and Drug Administration) guidelines and ADMET (Adsorption, Distribution, Metabolism, Excretion, Toxicology) properties. "

Made available in DSpace on 2014-06-11T19:31:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-12Bitstream added on 2014-06-13T19:01:46Z : No. of bitstreams: 1 graciani_apb_dr_botfm.pdf: 427562 bytes, checksum: b481f2e6f558c5bae2f08ca356c1ed60 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / Paracoccidioides brasiliensis (Pb), agente etiológico da paracoccidioidomicose, é um fungo dimórfico que sobrevive no interior de monócitos/macrófagos humanos não ativados. Estudos anteriores em nosso laboratório têm demonstrado que os monócitos humanos não ativados são incapazes de realizar atividade fungicida, e esse processo está associado com a capacidade do fungo induzir a produção de prostaglandinas (PGs), uma vez que, essas células são capazes de realizar atividade fungicida significativa após o tratamento com indometacina (INDO), um inibidor da produção de ciclooxigenase. No entanto, o processo de pré-ativação com IFN-γ, resulta em um parcial efeito compensatório sobre os efeitos inibidores das PGs, principalmente quando essas células são desafiadas com a cepa de baixa virulência do fungo. Assim, a proposta deste presente estudo foi avaliar se a ativação de monócitos humanos com outras citocinas como TNF-α e GM-CSF resulta em um efeito similar ao observado com IFN-γ. Uma outra questão a ser respondida é se esse processo poderia estar associado com alterações nos níveis de H2O2 e NO, que são moléculas efetoras envolvidas na atividade fungicida contra o P. brasiliensis, bem como nos níveis das citocinas TNF-α, IL-10 e IL-6. Culturas de monócitos do sangue periférico, obtidos de 20 indivíduos normais foram tratadas somente com INDO ou ativados com IFN-γ, TNF-α ou GM-CSF na presença ou ausência de INDO por 18h, e posteriormente desafiados com cepas de alta (Pb18) ou baixa (Pb265) virulência do P. brasiliensis por 4h. Após esse período, as culturas foram avaliadas quanto à atividade fungicida, produção de H2O2 e NO e expressão de mRNA para enzima óxido nítrico sintase (iNOS) por RT-PCR em tempo real. As concentrações de TNF-α, IL-6 e IL-10 nos sobrenadantes das coculturas foram avaliadas por ELISA. Nossos resultados... / Paracoccidioides brasiliensis (Pb), the etiological agent of paracoccidioidomycosis, is a dimorphic fungus that survives within nonactivated human monocytes/macrophages. Previous studies have demonstrated that the lack of fungicidal activity by nonactivated human monocytes is associated to fungus capacity to inducing prostaglandins release, since a significative fungicidal activity was detected after monocytes treatment with indomethacin (INDO), a cyclooxigenase inhibitor. However, cells activation with IFN-γ seems to partially compensating this inhibitory effect, mainly when cells were challenged with low virulent strain of the fungus. Here, we extended our studies, addressing whether monocytes activation with other cytokines such as TNF-α and GM-CSF results in a similar effect to that observed with IFN-γ. Moreover, we asked if this process could be associated with alterations on H2O2 and NO levels, the molecules involved in Pb killing, as well as in the levels of the cytokines TNF-α, IL-10 and IL-6. Peripheral blood monocytes obtained from 18 healthy donors were treated only with INDO or activated with IFN-γ, TNF-α or GM-CSF in presence or absence of INDO for 18h, and further challenged with high (Pb18) or low (Pb265) virulent strain of Pb for 4h. After, cultures were evaluated for fungicidal activity, H2O2 and NO release and expression of inducible nitric oxide synthase (iNOS) mRNA by real-time RT-PCR. The concentrations of TNF-α, IL-6 and IL-10 on supernatants of cocultures were evaluated by ELISA. Our results provided evidence that human monocytes challenged with both strains of P. brasiliensis release prostaglandins that via induction of IL-10 and IL-6 inhibits TNF-α production. This process results in defective cell activation with consequent release of low H2O2 levels and lack of fungicidal activity by cells. However the inhibitory effect of PGs may be... (Complete abstract click electronic access below)

The history of cultivating barley dates back to the beginning of conscious agriculture, accompanying humans together with wheat, as the second oldest grain. It is not possible to understand the current role barley plays in our economy only from the perspective of its use of malting, although we regard it as a priority, both in light of breeding and cultivating, and its research. With regard to the short growing season, the weaker root system and its biological character, barley reacts sensitive to stressful conditions of all kind, and thus also to any br1eeding mistake. Throughout its entire vegetation period, spring barley is exposed to the pressure of a number of fungal diseases. Especially in regard to the malting barley, it is necessary to fight against this danger effectively. High demands are put on the quality of malting barley and diseases can - to a large extent - decide on economic breeding results. It says: For successful breeding of malting barley, its outcome is the tax of the complex breeding proceedings. The fight against fungal diseases is a significant part of agricultural engineering. A series of preparations against fungal diseases exist and further registrations of preparations continue to occur. The laboratory Kluky s.r.o. is one of the companies that makes registered experiments of the preparations. Within this recognized laboratory, the experiment of monitoring diseases of the spring barley was carried out while applying chosen beneficent substances.

Resumo: Paracoccidioides brasiliensis (Pb), agente etiológico da paracoccidioidomicose, é um fungo dimórfico que sobrevive no interior de monócitos/macrófagos humanos não ativados. Estudos anteriores em nosso laboratório têm demonstrado que os monócitos humanos não ativados são incapazes de realizar atividade fungicida, e esse processo está associado com a capacidade do fungo induzir a produção de prostaglandinas (PGs), uma vez que, essas células são capazes de realizar atividade fungicida significativa após o tratamento com indometacina (INDO), um inibidor da produção de ciclooxigenase. No entanto, o processo de pré-ativação com IFN-γ, resulta em um parcial efeito compensatório sobre os efeitos inibidores das PGs, principalmente quando essas células são desafiadas com a cepa de baixa virulência do fungo. Assim, a proposta deste presente estudo foi avaliar se a ativação de monócitos humanos com outras citocinas como TNF-α e GM-CSF resulta em um efeito similar ao observado com IFN-γ. Uma outra questão a ser respondida é se esse processo poderia estar associado com alterações nos níveis de H2O2 e NO, que são moléculas efetoras envolvidas na atividade fungicida contra o P. brasiliensis, bem como nos níveis das citocinas TNF-α, IL-10 e IL-6. Culturas de monócitos do sangue periférico, obtidos de 20 indivíduos normais foram tratadas somente com INDO ou ativados com IFN-γ, TNF-α ou GM-CSF na presença ou ausência de INDO por 18h, e posteriormente desafiados com cepas de alta (Pb18) ou baixa (Pb265) virulência do P. brasiliensis por 4h. Após esse período, as culturas foram avaliadas quanto à atividade fungicida, produção de H2O2 e NO e expressão de mRNA para enzima óxido nítrico sintase (iNOS) por RT-PCR em tempo real. As concentrações de TNF-α, IL-6 e IL-10 nos sobrenadantes das coculturas foram avaliadas por ELISA. Nossos resultados... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Paracoccidioides brasiliensis (Pb), the etiological agent of paracoccidioidomycosis, is a dimorphic fungus that survives within nonactivated human monocytes/macrophages. Previous studies have demonstrated that the lack of fungicidal activity by nonactivated human monocytes is associated to fungus capacity to inducing prostaglandins release, since a significative fungicidal activity was detected after monocytes treatment with indomethacin (INDO), a cyclooxigenase inhibitor. However, cells activation with IFN-γ seems to partially compensating this inhibitory effect, mainly when cells were challenged with low virulent strain of the fungus. Here, we extended our studies, addressing whether monocytes activation with other cytokines such as TNF-α and GM-CSF results in a similar effect to that observed with IFN-γ. Moreover, we asked if this process could be associated with alterations on H2O2 and NO levels, the molecules involved in Pb killing, as well as in the levels of the cytokines TNF-α, IL-10 and IL-6. Peripheral blood monocytes obtained from 18 healthy donors were treated only with INDO or activated with IFN-γ, TNF-α or GM-CSF in presence or absence of INDO for 18h, and further challenged with high (Pb18) or low (Pb265) virulent strain of Pb for 4h. After, cultures were evaluated for fungicidal activity, H2O2 and NO release and expression of inducible nitric oxide synthase (iNOS) mRNA by real-time RT-PCR. The concentrations of TNF-α, IL-6 and IL-10 on supernatants of cocultures were evaluated by ELISA. Our results provided evidence that human monocytes challenged with both strains of P. brasiliensis release prostaglandins that via induction of IL-10 and IL-6 inhibits TNF-α production. This process results in defective cell activation with consequent release of low H2O2 levels and lack of fungicidal activity by cells. However the inhibitory effect of PGs may be... (Complete abstract click electronic access below) / Orientador: Ângela Maria Victoriano de Campos Soares / Coorientador: Luciene Alarcão Dias Melicio / Banca: Maria Terezinha Serrão Peraçoli / Banca: João pessoa Araújo Junior / Banca: Heloisa Blotta / Banca: Gil Bernard / Doutor

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Sementes de Platypodium elegans Vogel, pertencente à FamÃlia Fabaceae, subfamÃlia Papilionoideae, Tribo Dalbergiae possui uma lectina Manose/Glicose ligante. Na purificaÃÃo da lectina em estudo, o extrato total de sementes de P. elegans preparado em NaCl 0,15M teve sua fraÃÃo hemaglutinante isolada por cromatografia de afinidade em quitina, trocas iÃnica em DEAE-Sephacel e em HiTrap SP, essa Ãltima acoplada a HPLC. As fraÃÃes ativas para atividade hemaglutinante obtidas sequencialmente nesses procedimentos foram avaliados quanto à sua atividade especÃfica e homogeneidade em SDS-PAGE. A lectina em seu Ãltimo passo de purificaÃÃo apresentou massa molecular aparente de 55 kDa. A fraÃÃo prÃ-purificada ativa obtida em DEAE-Sephacel foi caracterizada quanto ao seu potencial termiticida e fungicida. A atividade termiticida contra operÃrios e soldados de Nasutitermes corniger, induzindo 100% de mortalidade em ambas as classes, foram melhor visualizadas com as concentraÃÃes 1,0 e 0,8mg/mL, entre 8 e 9 dias para operÃrios e 7 e 8 dias para soldados, onde ainda nÃo se havia registrado menos que 50% de mortalidade no controle negativo. A fraÃÃo testada nÃo apresentou efeito repelente. Em relaÃÃo à atividade fungicida, 20 ug da fraÃÃo chegou a inibir consideravelmente Fusarium lateritium (33%), F. oxysporum (19,4%) e F. solani (14,3%). F. decemcellulare teve uma inibiÃÃo de 4,8% e F. moniliforme, 3,3%. Essas duas atividades reforÃam a participaÃÃo dessa lectina na defesa vegetal, fazendo-a uma potente ferramenta biotecnolÃgica a ser mais profundamente investigada e estudada em relaÃÃo a seu papel na defesa vegetal e na potencial utilizaÃÃo na resistÃncia de madeiras susceptÃveis a cupins. / Seeds of Platypodium elegans Vogel, belonging to the Fabaceae family, Papilionoideae sub-family, Dalbergiae Tribe, posses a specific mannose/glucose lectin. In the purification of the lectin in study, the total extract of seeds of P. elegans prepared in NaCl 0,15M had its active fraction isolated for affinity chromatography in chitin and exchange ionic chromatography in DEAE-Sephacel and HiTrap SP, this last one connected to the HPLC.The active fractions for hemmagutinant activity gotten in these procedures had been sequentially about its specific activity and homogeneity in SDS-PAGE. The lectin in the last step of purification presented apparent molecular mass of 55 kDa. The active fraction from DEAE-Sephacel was characterized about its termiticidal and fungicidal potential. The termiticidal activity against laborers and soldiers of Nasutitermes corniger, inducing 100% of mortality in both the classrooms, was better visualized with the concentrations of 1,0 and 0,8mg/mL, between 8 and 9 days for laborers and 7 and 8 days for soldiers, where it had not been registered mortality less than 50% in the negative control. The tested fraction did not present repellent effect. In relation to the fungicidal activity, 20 ug of the fraction arrived to inhibit considerably Fusarium lateritium (33%), F. oxysporum (19.4%) and F. solani (14.3%). F. decemcellulare had a inhibition of 4,8% and F. moniliforme, 3.3%. These two activities strengthen the participation of this lectin in the vegetal defense, making it a powerful biotechnological tool to be more deeply investigated and studied about its paper in the vegetal defense and the potential use in the resistance of susceptible wood to termites.

<p> </p> <p>Disinfectant application on environmental surfaces is generally accepted to be effective</p> <p>against vegetative pathogens. However, several important application considerations are not being evaluated</p> <p>in the registration of these disinfectant solutions. This thesis looks into factors impacting the disinfectant wipes' bactericidal and fungicidal efficacy on hard, non-porous surfaces and how the interaction between the disinfectant solution and wipe substrate lead to changes in the efficacy of disinfection.</p>

Made available in DSpace on 2016-12-23T14:37:30Z (GMT). No. of bitstreams: 1 Tatiane Paulino da Cruz.pdf: 1055660 bytes, checksum: 58a56273fc2f252688911c2700a070e5 (MD5) Previous issue date: 2013-02-20 / The decline of guava has been responsible for the eradication of many orchards nationwide, resulting in high economic loss to producers. Due to increasing pressure from society for healthier products, free from chemicals and to increase the importance of essential oils in the management of plant diseases, the aim of this study was to determine the chemical composition and fungitoxic evaluate the effects of essential oils of Eucalyptus citriodora (eucalyptus), E. grandis (eucalyptus), Cymbopogon citratus (lemon grass), Cymbopogon winterianus (lemongrass), Cinnamomum zeylanicum (Cinnamon), Eucalyptus commercial Rosmarinos officinalis (rosemary), Ocimum gratissimum (basil-clove) and Syzygium aromaticum (clove india ) on three isolates of Fusarium solani and M. enterolobii. The essential oil was obtained by hydrodistillation using a Clevenger type apparatus, and its yield was determined relative to the dry weight of the plant. The analysis by gas chromatography and gas chromatography with mass detector of essential oils led to the identification of compounds. To evaluate the effect of essential oils on mycelial growth, sporulation and germination of spores of the fungus were used aliquots of 5, 10, 15, 20, 25 and 30 &#956;l of essential oils that were distributed on the surface of PDA culture medium contained in Petri dishes before subculturing the fungus. To evaluate the effect of oil on hatching and mortality of second stage juveniles (J2) of the nematode were used aqueous suspensions containing 200 eggs or 200 J2 which were added in the same rates of essential oils from botanicals mencinadas the previously. The experimental design was completely randomized with a 9 x 6 + 1, (nine oils, six aliquots and an additional control) with 5 repetitions. The lowest rate of essential oils of clove, cinnamon and clove basil-inhibited by more than 90% spore germination, mycelial growth and sporulation of the fungus isolates as well as the emergence of J2, as for oils eucalyptus, lemongrass, citronella inhibition was greater than 90% from the rate of 15 &#956;l, rosemary oil showed the worst result in 50% inhibiting the development of the fungus isolates / O declínio da goiabeira tem sido responsável pela erradicação de muitos pomares em âmbito nacional, o que resulta em elevada perda econômica aos produtores. Devido a crescente pressão da sociedade por produtos mais saudáveis, livres de produtos químicos e com aumento da importância dos óleos essenciais no manejo de doenças de plantas, objetivou-se com este trabalho determinar a composição química e avaliar os efeitos fungitóxicos dos óleos essenciais de Eucalyptus citriodora (eucalipto), E. grandis (eucalipto), Cymbopogon citratus (capim-limão), Cymbopogon winterianus (citronela), Cinnamomum zeylanicum (Canela), Eucalipto comercial, Rosmarinos officinalis (alecrim), Ocimum gratissimum (alfavaca-cravo) e Syzygium aromaticum (cravo-da-india) sobre três isolados de Fusarium solani e M. enterolobii. O óleo essencial foi obtido por hidrodestilação, utilizando um aparelho tipo Clevenger, e o seu rendimento foi determinado em relação à massa seca da planta. As análises de cromatografia gasosa e cromatografia gasosa com detector de massas de óleos essenciais levaram à identificação dos compostos. Para avaliar o efeito dos óleos essenciais no crescimento micelial, esporulação e germinação de esporos do fungo, foram utilizadas alíquotas de 5; 10; 15; 20; 25 e 30 &#956;L dos óleos essenciais que foram distribuídas na superfície do meio de cultura BDA contido em placas de Petri antes da repicagem do fungo. Para a avaliação do efeito dos óleos sobre a eclosão e mortalidade de juvenis de segundo estádio (J2) do nematóide foram utilizadas suspensões aquosas contendo 200 ovos ou 200 J2 nas quais foram adicionadas as mesmas alíquotas dos óleos essenciais das espécies botânicas mencinadas anteirormente. O delineamento experimental foi inteiramente casualizado num esquema fatorial 9 x 6 + 1, (nove óleos, seis aliquotas e uma testemunha adicional) com 5 repetições. A menor alíquota dos óleos essenciais de cravo-da-índia, canela e alfavaca-cravo inibiram em mais de 90% a germinação de esporos, o crescimento micelial e a esporulação dos isolados do fungo assim como a eclosão de J2, já para os óleos de eucalipto, capim limão, citronela a inibição foi superior a 90% a partir da alíquota de 15 &#956;l, o óleo de alecrim foi o que apresentou o pior resultado inibindo em 50% o desenvolvimento dos isolados do fungo

O desenvolvimento de resistência antimicrobiana e a consequente seleção de microrganismos multirresistentes consolidam-se como grandes ameaças à saúde global. Neste contexto, a busca por novas drogas antimicrobianas/microbicidas é fundamental e compostos como os peptídeos antimicrobianos (AMPs) tornaram-se alvos atraentes. Os AMPs são compostos químicos de massa molar média e grande diversidade estrutural, produzidos por todos os seres vivos e com capacidade de inibir o crescimento de e/ou matar microrganismos. O AMP Cheferina I (Chef I) foi isolado das raízes de Capsella bursa-pastoris e é resultado da proteólise de uma proteína da família das proteínas ricas em glicina, que em plantas estão relacionadas às funções de defesa e cicatrização. O nosso grupo de pesquisa foi pioneiro no desenvolvimento e estudo de análogos truncados amidados deste AMP atípico rico em glicina (67,9%) e histidina (28,6%), que se mostraram ativos frente às diferentes cepas de Candida e a S. cerevisiae pela internalização/ação celular acompanhada de manutenção da integridade da membrana plasmática; o análogo amidado (Chef Ia) e o análogo marcado com 5(6)-carboxifluoresceína/FAM (FAM-Chef Ia) tiveram as suas atividades antifúngicas potencializadas por íons Zn2+. Este trabalho deu continuidade ao estudo do efeito dos íons metálicos divalentes Zn2+, Cu2+, Ca2+ e Mg2+ nas atividades anticandida/fungistática e candidacida/fungicida a diferentes pHs e forças iônicas, estruturas e localizações intracelulares destes análogos. Os resultados na ausência de íons em pH 5,1 revelaram maior atividade do análogo fluorescente em relação à do não fluorescente. Neste mesmo pH, as atividades anticandida e candidacida de Chef Ia foram influenciadas negativamente pelos íons Ca2+ e Mg2+ (2-4 vezes) enquanto que, na presença de íons Zn2+ as atividades anticandida de ambos os análogos foram aumentadas (Chef Ia: 8-64 vezes; FAM-Chef Ia: 4-32 vezes). Os íons Cu2+ aumentaram a atividade anticandida de Chef Ia (2-4 vezes), mas não a do análogo fluorescente, mas as atividades candidacidas de ambos foram melhoradas (Chef Ia: 2-8 vezes; FAM-Chef Ia: 2 vezes). Em pH 5,1, os íons Zn2+ mantiveram a atividade anticandida de Chef Ia em alta força iônica, mas só FAM-Chef Ia exibiu atividade candidacida. Em pH 7,4 ambos análogos foram inativos em baixa e alta forças iônicas na ausência e presença de Zn2+ ou Cu2+. As maiores porcentagens de folhas-&#946;-antiparalelas e dobras foram observadas no espectro de DC de Chef Ia em pH 7,4, sendo que aqueles registrados em pH 5,1 e 7,4 em presença de íons Zn2 e Cu2+ indicaram a formação de quelatos estruturalmente distintos. Ambos os peptídeos são bioquelantes em potencial, sendo as proporções peptídeo: íon obtidas as seguintes: FAM-Chef Ia = 1:2 para Cu2+, 1:10 para Zn2+; Chef Ia = 1:1 para Cu2+. A análise da internalização celular de FAM-Chef Ia permitiu a suposição de dois mecanismos de internalização (translocação direta e endocitose), sendo que nas células vivas a presença de Zn2+ afetou negativamente a translocação direta (p 0,0343) e potencializou a endocitose (p 0,0002). / The development of antimicrobial resistance and the consequent selection of multiresistant microorganisms have become major threats to global health. In this context, the search for new antimicrobial/microbicidal drugs is crucial and the antimicrobial peptides (AMPs) have been seen as attractive targets. AMPs are chemical compounds of medium molecular mass and high structural diversity produced by all living beings, capable of inhibiting the growth of microorganisms and killing them. The AMP Shepherin I (Shep I) was isolated from the roots of Capsella bursa-pastoris, being a bioactive peptide encrypted in a glycine-rich protein from a family that in plants are strictly related to defense and healing functions. Our research group has pioneered the development and study of amidated truncated analogues of this atypical glycine- (67.9%) and histidine-rich (28.6%) AMP, which has shown activity against different strains of Candida and S. cerevisiae through cellular internalization with maintenance of the plasma membrane integrity. The amide analogue (Chef Ia) and its fluorescent analog labeled with 5 (6) - carboxyfluorescein / FAM (FAM-Chef Ia) had their antifungal activities potentiated by Zn2+ ions, so the present work continued examining the effect of the divalent metallic ions Zn2+, Cu2+, Ca2+ and Mg2+ on the anticandidal/fungistatic and candidacidal/fungicide activities at different pHs and ionic forces, structures and intracellular locations of these analogues. The results in the absence of those ions at pH 5.1 revealed that the fluorescently labelled analog was more potent than the nonfluorescent. At the same pH, Shep Ia anticandidal and candidacidal activities were negatively influenced by Ca2+ and Mg2+ ions (2-4 fold), whereas in the presence of Zn2+ ions the anticandidal activities of both analogues were increased (Shep Ia: 8-64 fold, FAM- Shep Ia: 4-32 fold). Cu2+ ions increased Shep Ia anticandidal activity (2-4 fold) but not that of FAM-Shep Ia, nevertheless, the candidacidal activities of both analogues were increased (Shep Ia: 2-8 fold, FAM-Shep Ia: 2 fold). Also at pH 5.1, the Zn2+ ions helped retaining the anticandidal activity of Shep Ia at high ionic strength, although only FAM-Shep Ia exhibited candidacidal activity. At pH 7.4 both analogues were inactive at low and high ionic strengths in the absence or presence of Zn2+ or Cu2+. The highest percentages of antiparallel &#946;-sheet and turns were observed in Shep Ia CD spectrum at pH 7.4, while those recorded at pH 5.1 and 7.4 in the presence of Zn2+ or Cu2+ ions indicated the formation of structurally different chelates. Both peptides are potential biochelates, with the following peptide:ion ratios: FAM-Shep Ia = 1: 2 for Cu2+, 1:10 for Zn2+; Shep Ia = 1: 1 for Cu2+. The analysis of the cellular internalization of FAM-Chef Ia allowed the assumption of two mechanisms of internalization (direct translocation and endocytosis) and in the living cells the presence of Zn2+ negatively affected the direct translocation (p 0.0343) and potentiated endocytosis (p 0.0002).

BENEVIDES, Raquel Guimarães. Avaliação do potencial fungicida e termiticida de uma fração protéica lectínica de sementes de Platypodium elegans Vogel e obtenção da lectina purificada. 2008. 113 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal do Ceará, Fortaleza-CE, 2008. / Submitted by Eric Santiago (erichhcl@gmail.com) on 2016-07-04T12:43:49Z No. of bitstreams: 1 2008_dis_rgbenevides.pdf: 5092516 bytes, checksum: 86289fcb11e2cb709d80b92c6f7cd851 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-08-02T20:15:19Z (GMT) No. of bitstreams: 1 2008_dis_rgbenevides.pdf: 5092516 bytes, checksum: 86289fcb11e2cb709d80b92c6f7cd851 (MD5) / Made available in DSpace on 2016-08-02T20:15:19Z (GMT). No. of bitstreams: 1 2008_dis_rgbenevides.pdf: 5092516 bytes, checksum: 86289fcb11e2cb709d80b92c6f7cd851 (MD5) Previous issue date: 2008 / Seeds of Platypodium elegans Vogel, belonging to the Fabaceae family, Papilionoideae sub-family, Dalbergiae Tribe, posses a specific mannose/glucose lectin. In the purification of the lectin in study, the total extract of seeds of P. elegans prepared in NaCl 0,15M had its active fraction isolated for affinity chromatography in chitin and exchange ionic chromatography in DEAE-Sephacel and HiTrap SP, this last one connected to the HPLC.The active fractions for hemmagutinant activity gotten in these procedures had been sequentially about its specific activity and homogeneity in SDS-PAGE. The lectin in the last step of purification presented apparent molecular mass of 55 kDa. The active fraction from DEAE-Sephacel was characterized about its termiticidal and fungicidal potential. The termiticidal activity against laborers and soldiers of Nasutitermes corniger, inducing 100% of mortality in both the classrooms, was better visualized with the concentrations of 1,0 and 0,8mg/mL, between 8 and 9 days for laborers and 7 and 8 days for soldiers, where it had not been registered mortality less than 50% in the negative control. The tested fraction did not present repellent effect. In relation to the fungicidal activity, 20 ug of the fraction arrived to inhibit considerably Fusarium lateritium (33%), F. oxysporum (19.4%) and F. solani (14.3%). F. decemcellulare had a inhibition of 4,8% and F. moniliforme, 3.3%. These two activities strengthen the participation of this lectin in the vegetal defense, making it a powerful biotechnological tool to be more deeply investigated and studied about its paper in the vegetal defense and the potential use in the resistance of susceptible wood to termites. / Sementes de Platypodium elegans Vogel, pertencente à Família Fabaceae, subfamília Papilionoideae, Tribo Dalbergiae possui uma lectina Manose/Glicose ligante. Na purificação da lectina em estudo, o extrato total de sementes de P. elegans preparado em NaCl 0,15M teve sua fração hemaglutinante isolada por cromatografia de afinidade em quitina, trocas iônica em DEAE-Sephacel e em HiTrap SP, essa última acoplada a HPLC. As frações ativas para atividade hemaglutinante obtidas sequencialmente nesses procedimentos foram avaliados quanto à sua atividade específica e homogeneidade em SDS-PAGE. A lectina em seu último passo de purificação apresentou massa molecular aparente de 55 kDa. A fração pré-purificada ativa obtida em DEAE-Sephacel foi caracterizada quanto ao seu potencial termiticida e fungicida. A atividade termiticida contra operários e soldados de Nasutitermes corniger, induzindo 100% de mortalidade em ambas as classes, foram melhor visualizadas com as concentrações 1,0 e 0,8mg/mL, entre 8 e 9 dias para operários e 7 e 8 dias para soldados, onde ainda não se havia registrado menos que 50% de mortalidade no controle negativo. A fração testada não apresentou efeito repelente. Em relação à atividade fungicida, 20 ug da fração chegou a inibir consideravelmente Fusarium lateritium (33%), F. oxysporum (19,4%) e F. solani (14,3%). F. decemcellulare teve uma inibição de 4,8% e F. moniliforme, 3,3%. Essas duas atividades reforçam a participação dessa lectina na defesa vegetal, fazendo-a uma potente ferramenta biotecnológica a ser mais profundamente investigada e estudada em relação a seu papel na defesa vegetal e na potencial utilização na resistência de madeiras susceptíveis a cupins.