Worldwide occurrence of feline hemoplasma infections in wild felid species

While hemoplasma infections in domestic cats are well studied, almost no information is available on their occurrence in wild felids. The aims of the present study were to investigate wild felid species as possible reservoirs of feline hemoplasmas and the molecular characterization of the hemoplasma isolates. Blood samples from the following 257 wild felids were analyzed: 35 Iberian lynxes from Spain, 36 Eurasian lynxes from Switzerland, 31 European wildcats from France, 45 lions from Tanzania, and 110 Brazilian wild felids, including 12 wild felid species kept in zoos and one free-ranging ocelot. Using real-time PCR, feline hemoplasmas were detected in samples of the following species: Iberian lynx, Eurasian lynx, European wildcat, lion, puma, oncilla, Geoffroy's cat, margay, and ocelot. "Candidatus Mycoplasma haemominutum" was the most common feline hemoplasma in Iberian lynxes, Eurasian lynxes, Serengeti lions, and Brazilian wild felids, whereas "Candidatus Mycoplasma turicensis" was the most prevalent in European wildcats; hemoplasma coinfections were frequently observed. Hemoplasma infection was associated with species and free-ranging status of the felids in all animals and with feline leukemia virus provirus-positive status in European wildcats. Phylogenetic analyses of the 16S rRNA and the partial RNase P gene revealed that most hemoplasma isolates exhibit high sequence identities to domestic cat-derived isolates, although some isolates form different subclusters within the phylogenetic tree. In conclusion, 9 out of 15 wild felid species from three different continents were found to be infected with feline hemoplasmas. The effect of feline hemoplasma infections on wild felid populations needs to be further investigated

Síntese e caracterização de arcabouços de quitosana com agente antineoplásicos

O sistema de liberação controlada de fármacos através da utilização de biomateriais poliméricos associados a compostos com ação antineoplásica pode ser empregado como alternativa de tratamento de neoplasias. Desta forma, este trabalho teve como objetivo a síntese e caracterização de sistemas de arcabouços de quitosana com o agente antineoplásico (1,4-naftoquinona), cuja taxa de liberação pode ser controlada pela utilização de um agente reticulante como o tripolifosfato de sódio (TPP). O método de preparação consistiu da solubilização da quitosana em ácido acético, adição do fármaco, congelamento, liofilização e reticulação com TPP. Todas as amostras foram caracterizadas por Difração de Raios X (DRX), Microscopia Eletrônica de Varredura (MEV), Espectroscopia de Energia Dispersiva de Raios X(EDS), grau de intumescimento e biodegradação enzimática. Na MEV foi evidenciada a formação de poros interconectados com tamanhos e formas variadas em todas as estruturas estudadas caracterizando a formação de arcabouços. Já no EDS foi observada a presença de elementos químicos característico da composição química de cada material. No entanto foi observada a presença do sódio que pode estar relacionado ao agente neutralizante utilizado. A reticulação de parte dos arcabouços foi comprovada pelo DRX, EDS e aumentou a taxa de degradação enzimática in vitro dos mesmos. A incorporação do fármaco foi confirmada por DRX, grau de intumescimento e EDS. Desta forma, pode-se concluir que ocorreu à formação de arcabouços reticulados e não reticulados porosos, com propriedades morfológicas e físico-químicas que podem contribuir para carrear fármacos antineoplásicos, sendo possível controlar a taxa de degradação dos mesmos e provável liberação do fármaco

Evaluation of the Allergenicity Potential of TcPR-10 Protein from Theobroma cacao

Background: The pathogenesis related protein PR10 (TcPR-10), obtained from the Theobroma cacao-Moniliophthora perniciosa interaction library, presents antifungal activity against M. perniciosa and acts in vitro as a ribonuclease. However, despite its biotechnological potential, the TcPR-10 has the P-loop motif similar to those of some allergenic proteins such as Bet v 1 (Betula verrucosa) and Pru av 1 (Prunus avium). The insertion of mutations in this motif can produce proteins with reduced allergenic power. The objective of the present work was to evaluate the allergenic potential of the wild type and mutant recombinant TcPR-10 using bioinformatics tools and immunological assays. Methodology/Principal Findings: Mutant substitutions (T10P, I30V, H45S) were inserted in the TcPR-10 gene by sitedirected mutagenesis, cloned into pET28a and expressed in Escherichia coli BL21(DE3) cells. Changes in molecular surface caused by the mutant substitutions was evaluated by comparative protein modeling using the three-dimensional structure of the major cherry allergen, Pru av 1 as a template. The immunological assays were carried out in 8-12 week old female BALB/c mice. The mice were sensitized with the proteins (wild type and mutants) via subcutaneous and challenged intranasal for induction of allergic airway inflammation. Conclusions/Significance: We showed that the wild TcPR-10 protein has allergenic potential, whereas the insertion of mutations produced proteins with reduced capacity of IgE production and cellular infiltration in the lungs. On the other hand, in vitro assays show that the TcPR-10 mutants still present antifungal and ribonuclease activity against M. perniciosa RNA. In conclusion, the mutant proteins present less allergenic potential than the wild TcPR-10, without the loss of interesting biotechnological properties. (Résumé d'auteur