Therapeutic switching: from antidermatophytic essential oils to new leishmanicidal products

This study examined whether the antidermatophytic activity of essential oils (EOs) can be used as an indicator for the discovery of active natural products against Leishmania amazonensis. The aerial parts of seven plants were hydrodistilled. Using broth microdilution techniques, the obtained EOs were tested against three strains of dermatophytes (Trichophyton mentagrophytes, Microsporum gypseum and Microsporum canis). To compare the EOs antifungal and antiparasitic effects, the EOs activities against axenic amastigotes of L. amazonensis were concurrently evaluated. For the most promising EOs, their antileishmanial activities against parasites infecting peritoneal macrophages of BALB/c mice were measured. The most interesting antifungal candidates were the EOs from Cymbopogon citratus, Otacanthus azureus and Protium heptaphyllum, whereas O. azureus, Piper hispidum and P. heptaphyllum EOs exhibited the lowest 50% inhibitory concentration (IC50) values against axenic amastigotes, thus revealing a certain correspondence between both activities. The P. hispidum EO was identified as the most promising product in the results from the infected macrophages model (IC50: 4.7 µg/mL, safety index: 8). The most abundant compounds found in this EO were sesquiterpenes, notably curzerene and furanodiene. Eventually, the evaluation of the antidermatophytic activity of EOs appears to be an efficient method for identifying new potential drugs for the treatment of L. amazonensis.

Secreted proteases from dermatophytes

Dermatophytes are highly specialized pathogenic fungi that exclusively infect the stratum corneum, nails or hair, and it is evident that secreted proteolytic activity is important for their virulence. Endo- and exoproteases-secreted by dermatophytes are similar to those of species of the genus Aspergillus. However, in contrast to Aspergillus spp., dermatophyte-secreted endoproteases are multiple and are members of two large protein families, the subtilisins (serine proteases) and the fungalysins (metalloproteases). In addition, dermatophytes excrete sulphite as a reducing agent. In the presence of sulphite, disulphide bounds of the keratin substrate are directly cleaved to cysteine and S-sulphocysteine, and reduced proteins become accessible for further digestion by various endo- and exoproteases secreted by the fungi. Sulphitolysis is likely to be an essential step in the digestion of compact keratinized tissues which precedes the action of all proteases.

New and newly illustrated Gnomidolon Thomson (Coleoptera, Cerambycidae, Hexoplonini) from Bolivia

Three new species of Gnomidolon Thomson, 1864 (Coleoptera, Cerambycidae, Hexoplonini) from Bolivia are described and illustrated, and G. proseni Martins, 1962 is redefined. Photographs of G. proseni and four other Bolivian species of this genus are provided: G. bonsae Martins, 1967, G. nanum Martins, 1962, G. musivum (Erichson, 1847), and both pale and dark forms of G. wappesi Martins, 2006.

Cytotaxonomy of North African species of Delphinium L. sect. Delphinium (Ranunculaceae).

S'han estudiat els nombres cromosómics i l'estructura dels cariotipus de 15 poblacions nordafricanes de Delphinium L. sect. Delphinium. Delphinium balansae Boiss. & Reuter, amb cariotipus mes simétric, apareix com a possible ancestre perenne de la resta d'espécies annuals amb cariotipus mes asimétric i mes curt. Es proposa una reorganització taxonómica de la secció, d'on es descriuen dues series noves (ser. Macropeiala ser. nova i ser. Balansae ser. nova.) i es proposen dues combinacions noves (D. nanum subsp. alboliliaceum i D. nanum subsp. elongatum).

El programa Biodiversitat Micològica de les terres de Ponent. Notícia i primers resultats.

La tardor de 1994 ha presentat unes condicions molt favorables de pluviositat, humitat i temperatura, que han permès un estudi intensiu (59 excursions, 64 localitats) de la part occidental, baixa i seca, de Catalunya, de la qual es tenien molt poques dades micològiques. E1 catàleg preliminar que oferim, amb tot i l'absència de moltes espècies pendents d'estudi o confirmació, i també de les més eurioiques, considerades poc característiques, conté 170 espècies, i permet fer-se una idea de la flora fúngica xero-termòfila mediterrània, especialment, la de les brolles, pinedes i espais oberts. Al costat d'algunes espècies poc citades, com Eutryblidiella hysterina, Helvella villosa, Agaricus pilatianus, Amanita boudieri, Calyptella capula, Ceriporia bresadolae, Coprinus vosoustii, Henningsomyces puber, Hygrophorus carneogriseus, Marasmius corbariensis, Phellinus punctatus, Ramicola iberica, Skeletocutis percandida, Tulostoma nanum, T. occidentale, T. xerophilum, Typhula setipes, Xerocomus ichnusanus, d'altres han mostrat una abundància inusitada, com Mycocalicium minutellum, Amanita ovoidea, Clitocybe alexandrí, C. umbilicata, Entoloma rusticoides, Hebeloma edurum, Inocybe roseipes, Lepista rickenii, Lopharia spadicea, Omphalotus olearius, Phaeotellus rickenii, Polyporus meridionalis, Suillus bellinii, S. collinitus, Volvariella speciosa, Mucilago crustacea. És previst continuar les prospeccions, per a completar aquesta primera visió.

Identification des dermatophytes causant des teignes de la peau glabre (Tinea glabra) et du cuir chevelu (Tinea capitis) par PCR

Contexte: Le nombre de teignes du cuir chevelu et de la peau glabre étant en nette augmentation, l'identification du pathogène qui est indispensable pour un traitement ciblé, a, par conséquence, un grand intérêt pour la santé publique. Dans divers cas, un animal de compagnie peut être identifié en tant que source du pathogène. La fréquence de cultures restant stériles est particulièrement élevée en cas de prétraitement antifongique. Objectif: Le but de ce travail est de mettre au point une méthode rapide d'identification du dermatophyte pathogène in situ par PCR/séquençage dans les cas de teignes du cuir chevelu et/ou de la peau glabre. Matériel et méthodes : De l'ADN a été extrait de squames (N=5) et cheveux (N=21) dont l'examen direct démontrait une infection fongique (N=26) ou se révèlait négatif (N=1). Ensuite, une première PCR du segment 28s de l'ADN ribosomale fongique a été effectuée, suivie par une PCR nichée intérieure à ce segment. L'amplicon a été séquencé et le champignon est identifié par alignement. Résultats : Seule la PCR enchainée a permis d'obtenir une quantité suffisante d'amplicon pour permettre le séquençage. Dans 4 cas sur 5 de tinea pedis, 10 sur 12 de tinea glabra, respectivement 4 sur 4 de tinea capitis, dans lesquels un dermato- phyte a été identifié en culture, le même dermatophyte a été identifié par PCR/séquençage. Une fois sur 27 prélèvements, un autre dermatophyte a été identifié par PCR/séquençage. Ce résultat pourrait être dû à une fausse identification du champignon en culture. Dans un cas de tinea pedis et un cas de tinea corporis, la culture est restée stérile, mais un dermatophyte a pu être identifié par PCR et séquençage. Conclusions : La méthode décrite est à la fois rapide (24 h au lieu de deux semaines pour la culture), sensible et très spécifique. Elle est particulièrement utile dans les cas de teigne du cuir chevelu, dans lesquels le traitement est différent selon l'espèce de dermatophyte et où il s'agit d'un traitement systémique lourd, souvent chez l'enfant.

Identification of novel secreted proteases during extracellular proteolysis by dermatophytes at acidic pH.

The dermatophytes are a group of closely related fungi which are responsible for the great majority of superficial mycoses in humans and animals. Among various potential virulence factors, their secreted proteolytic activity attracts a lot of attention. Most dermatophyte-secreted proteases which have so far been isolated in vitro are neutral or alkaline enzymes. However, inspection of the recently decoded dermatophyte genomes revealed many other hypothetical secreted proteases, in particular acidic proteases similar to those characterized in Aspergillus spp. The validation of such genome predictions instigated the present study on two dermatophyte species, Microsporum canis and Arthroderma benhamiae. Both fungi were found to grow well in a protein medium at acidic pH, accompanied by extracellular proteolysis. Shotgun MS analysis of secreted protein revealed fundamentally different protease profiles during fungal growth in acidic versus neutral pH conditions. Most notably, novel dermatophyte-secreted proteases were identified at acidic pH such as pepsins, sedolisins and acidic carboxypeptidases. Therefore, our results not only support genome predictions, but demonstrate for the first time the secretion of acidic proteases by dermatophytes. Our findings also suggest the existence of different pathways of protein degradation into amino acids and short peptides in these highly specialized pathogenic fungi.

CARACTERÍSTICAS FÍSICAS DE PEDÚNCULOS DE CAJUEIRO PARA COMERCIALIZAÇÃO IN NATURA

Com o objetivo de selecionar pedúnculos de cajueiro-anão precoce (Anacardium occidentale L. var. nanum) para comercialização in natura, foram avaliados 09 clones selecionados a partir de um experimento de competição sob irrigação, no município de Mossoró-RN. O clone CCP 76 foi utilizado como testemunha. Os cajus foram colhidos em agosto de 1997 e avaliados quanto às seguintes características: textura, tamanho (diâmetros apical e basal e comprimento), formato, coloração (carta de cores e pigmentos) e peso (total e pedúnculo). Dentre os materiais avaliados, apenas o CCP 09 apresentou cor inferior à testemunha, sendo que os clones CAP 6 (500), END 157, END 189 e END 329 destacaram-se com coloração mais intensa que a mesma. Além da testemunha, apenas os clones END 157, 183 e 189 apresentaram pedúnculos que podem ser classificados como tipo 4 (de maior valor comercial), enquanto, com relação à forma, apenas os clones CAP 6 (500), END 157 e END 183 apresentaram formato piriforme. O clone END 157 apresentou as melhores características para comercialização in natura, inclusive quando comparado à testemunha. Os clones END 183 e 189 apresentaram resultados semelhantes à testemunha, com exceção da cor para o 183 e do formato para o 189.

Glicoalcaloides antifúngicos, flavonoides e outros constituintes químicos de Solanum asperum

Two glycoalkaloids: solamargine and solasonine; three flavonoids: tiliroside, 7-O-α-L-ramnopyranosyl-kaempferol and 3-O-[ß-D-glucopyranosyl-(1→6)-α-L-ramnopyranosyl]-7- O-α-L-ramnopyranosyl-kaempferol, in addition to the tripeptide Leu-Ile-Val, the aminoacid proline and the eicosanoic acid were isolated from Solanum asperum (Solanaceae). The structures of all compounds were determined by interpretation of their spectra (IR, MS, ¹H and 13C NMR) and comparison with the literature data. All compounds, except the glycoalkaloids, are being reported for the first time for S. asperum. Solasonine showed strong activity (MIC < 0.24 μg/mL) against four filamentous fungi species of the genera Microsporum and Trichophyton.

FLAVONOIDES DE Piper glandulosissimum Yuncker (Piperaceae)

Three flavanones, two chalcones and one dihydrochalcone were isolated from the branches of Piper glandulosissimum. All isolated compounds were characterized based on IR, UV, 1H and 13C NMR, including 2D NMR analyses (HMQC, HMBC, COSY and NOESY) and comparison with the literature. The compound 7-hydroxy-5,8-dimethoxyflavanone displayed antimicrobial activity against Staphylococcus aureus, S. epidermidis, Trichophyton mentagrophytes and Microsporum canis.

Atividade antagônica a microrganismos patogênicos por bactérias endofíticas isoladas de Echinodorus scaber Rataj

Os objetivos desse trabalho foram identificar e avaliar o potencial antagônico in vitro de bactérias endofíticas isoladas de Echinodorus scaber (chapéu de couro) sobre alguns patógenos e verificar sua capacidade de controlar o desenvolvimento de fungos em grãos de soja. Um total de 113 linhagens foi confrontado com cinco fungos patogênicos (método de cultura dupla), e quatro bactérias patogênicas (método de sobrecamada). O controle de crescimento de fungo em grãos de soja foi realizado por microbiolização e avaliado pelo método de papel de filtro. As bactérias antagonistas foram submetidas a teste de antibiose contra quatro bactérias patógenas. Duas linhagens inibiram os fungos Colletotrichum lindemunthianum, C. gloeosporioides, Corynespora cassiicula, Fusarium solani, Microsporum canis. No teste de antibiose contra as bactérias patogênicas somente (BREIII-107) apresentou atividade antagônica. As duas linhagens e foram identificadas como Bacillus sp (BREI-92) e Bacillus subitilis (BREIII-107). Quando inoculadas em grãos de soja, Bacillus sp (BREI-92) e Bacillus subitilis (BREIII-107) inibiram aproximadamente 100% do desenvolvimento de fungos sobre os grãos.

Lufenuron no tratamento da dermatofitose em gatos?

Em função de controvérsias sobre a eficácia do lufenuron no tratamento da dermatofitose causada por Microsporum canis, o efeito da droga foi avaliado em 46 gatos (30 com lesões cutâneas e 16 portadores assintomáticos) atendidos no Hospital Veterinário da Universidade Federal Rural do Rio de Janeiro, Brasil. O diagnóstico foi estabelecido através da lâmpada de Wood, da cultura fúngica e, adicionalmente, na maioria dos animais sintomáticos, pela avaliação histopatológica da pele. Os animais foram tratados com 120mg/kg de lufenuron, a cada 21 dias (quatro doses); a droga mostrou-se eficaz no tratamento de 29 dos 30 felinos que apresentaram a forma clínica da dermatofitose, bem como no de todos os animais assintomáticos. O único felino que não teve cura clínica completa havia recebido várias doses de dexametasona antes do inicio do tratamento. Em um animal gestante, foram utilizadas duas doses do lufenuron e não foi observada qualquer alteração clínica e morfológica nos filhotes. Nenhum dos animais tratados teve qualquer reação adversa ao medicamento. Vinte dias após o último tratamento, o exame micológico resultou negativo em 45 dos felinos estudados (98%). Embora o lufenuron tenha o custo um pouco mais elevado do que o do cetoconazol, é uma droga que apresenta praticidade e margem de segurança bem maiores. Vale ressaltar que o sucesso do tratamento está diretamente relacionado à correta utilização da droga, assim como a perfeita higienização do ambiente e dos utensílios dos animais.

Ontogenia foliar de três espécies de Erythroxylum P. Browne (Erythroxylaceae) ocorrentes no Cerrado

Com o objetivo de descrever a ontogenia foliar e natureza das cicatrizes, encontradas em lâminas adultas de Erythroxylum, foram analisadas as folhas de Erythroxylum campestre, E. nanum e E. tortuosum, em diferentes estágios de desenvolvimento. Para as três espécies, verificou-se a ocorrência de um crescimento foliar marginal do tipo "submarginal mediano". No início do desenvolvimento do órgão, a atividade do meristema adaxial é intensa e leva à formação de uma saliência nessa face, enquanto que a porção laminar se desenvolve de forma involuta, devido à atividade do meristema marginal. Os estômatos se diferenciam junto com as camadas do mesofilo e a expansão final do órgão é dada por um crescimento intercalar. Durante a ontogenia não foi registrado o aparecimento de cicatrizes laminares que, quando presentes, se assemelham a injúrias superficiais, dispostas na face inferior e paralelamente à nervura principal.

Characterization and PCR multiplexing of polymorphic microsatellite loci in cashew (Anacardium occidentale L.) and their cross-species utilization

Cashew (Anacardium occidentale L.) is the most economically important tropical nut crop in the world, and yet there are no sequence tagged site (STS) markers available for its study. Here we use an automated, high-throughput system to isolate cashew microsatellites from a non-enriched genomic library blotted onto membranes at high density for screening. Sixty-five sequences contained a microsatellite array, of which 21 proved polymorphic among a closely related seed garden population of 49 genotypes. Twelve markers were suitable for multiplex analysis. Of these, 10 amplified in all three related tropical tree species tested: Anacardium microcarpum, Anacardium pumilum and Anacardium nanum.

Avaliação do sistema conservante em formulação com extrato hidroalcóolico de schinus terebinthifolius raddi nacardiaceae

Schinus terebinthifolius Raddi is used in the treatment of skin and mucosal injuries, infections of respiratory, digestive and genitourinary systems. Currently one of the biggest problems faced for the industry of phytopharmaceuticals with regard to the quality of raw materials is the microbial contamination. The aim this study was to evaluate the antimicrobial action of the hidroalcoholic extract of aroeira, beyond testing the effectiveness of the preservative system in hidrogel to the base of this extract. The extracts were prepared by maceration in the ratio of 1:10 of solvent plant/with alcohol 40%. The methods for microbial count were pour plate and test for specific microorganisms, analyzing in third copy each one of the samples. The antimicrobial activity of aroeira extracts was performed using an agar diffusion method, using strains of S. aureus, P. aeruginosa, E. coli, B. subtilis, C. albicans, C. tropicalis, C. krusei, C. guilliermondii, T. rubrum, M. gypseum, A. flavus and A. niger. The formula with aroeira was evaluated by the challenge test. This method consisted of artificial contamination the sample with separate inóculos of A. niger, C. albicans, E. coli e S. aureus aeruginosa and determinations of survivors for the method of counting for pour plate , during times 0, 24h, 48h, 7 days, 14 days, 21 days and 28 days. How much to the results, one verified that the extract of aroeira in the 13,5 concentration mg/mL presented antimicrobial activity for cepas of E. coli, B. subtilis, P. aeruginosa e S. aureus, producing inhibition zone, on average with 13 mm of diameter. However it did not present no fungi activity. The formula with aroeira containig both methylparaben and propylparaben showed a good efficacy in challenge test front to strains of A. niger, C. albicans, E. coli, S. aureus. The A criteria of European Pharmacopoeia, adopted in this work, was verified that this product revealed the good preservative efficacy for the challenge test, time interval of the 28 days. However, it is interesting to extend this study, in order to carry through the sped up stability and the test of shelf, to establish the validity of this formularization