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Abstract Objective For treatment of medication-related osteonecrosis of the jaw, one proposed approach is the use of a topical agent to block entry of these medications in oral soft tissues. We tested the ability of phosphonoformic acid (PFA), an inhibitor of bisphosphonate entry through certain sodium-dependent phosphate contransporters (SLC20A1, 20A2, 34A1-3) as well as Dynasore, a macropinocytosis inhibitor, for their abilities to prevent zoledronate-induced (ZOL) death in human gingival fibroblasts (HGFs). Methodology MTT assay dose-response curves were performed to determine non-cytotoxic levels of both PFA and Dynasore. In the presence of 50 μM ZOL, optimized PFA and Dynasore doses were tested for their ability to restore HGF viability. To determine SLC expression in HGFs, total HGF RNA was subjected to quantitative real-time RT-PCR. Confocal fluorescence microscopy was employed to see if Dynasore inhibited macropinocytotic HGF entry of AF647-ZOL. Endosomal acidification in the presence of Dynasore was measured by live cell imaging utilizing LysoSensor Green DND-189. As a further test of Dynasore's ability to interfere with ZOL-containing endosomal maturation, perinuclear localization of mature endosomes containing AF647-ZOL or TRITC-dextran as a control were assessed via confocal fluorescence microscopy with CellProfiler™ software analysis of the resulting photomicrographs. Results 0.5 mM PFA did not rescue HGFs from ZOL-induced viability loss at 72 hours while 10 and 30 μM geranylgeraniol did partially rescue. HGFs did not express the SLC transporters as compared to the expression in positive control tissues. 10 μM Dynasore completely prevented ZOL-induced viability loss. In the presence of Dynasore, AF647-ZOL and FITC-dextran co-localized in endosomes. Endosomal acidification was inhibited by Dynasore and perinuclear localization of both TRITC-dextran- and AF647-ZOL-containing endosomes was inhibited by 30 μM Dynasore. Conclusion Dynasore prevents ZOL-induced viability loss in HGFs by partially interfering with macropinocytosis and by inhibiting the endosomal maturation pathway thought to be needed for ZOL delivery to the cytoplasm.
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The genus Cerradoa (type species Cerradoa palmaea) was established in 1978 by Hennen and Ono and named after the Brazilian Cerrado biome. The holotype collected in Planaltina, Federal District, Brazil, belonged to the first rust fungus reported on palms (Arecaceae). For decades, the status of Cerradoa as a distinct genus has been regarded as doubtful, representing a synonym of Edythea (Uropyxidaceae) starting with the second edition of the Illustrated Genera of Rust Fungi in 1983. Our molecular phylogenetic analyses, as well as our morphological investigations, allowed us to reject this synonymy, leading to the reinstatement of Cerradoa within the Pucciniaceae. Cerradoa, together with morphologically similar genera such as the newly established Pseudocerradoa with two species (Ps. paullula and Ps. rhaphidophorae) infecting araceous hosts, the fern rust Desmella, and also P. engleriana, could not be assigned to any of the seven identified major lineages within the Pucciniaceae. Edythea, instead of being maintained as a member of the Uropyxidaceae, was herein placed in Pucciniaceae, shown phylogenetically in close relationship to Cumminsiella mirabilissima, both infecting the Berberidaceae. Additionally, our extensive phylogenetic analyses add guidance for future taxonomic revisions in the highly polyphyletic genus Puccinia and other established taxa within the family Pucciniaceae.
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Filogenia , BrasilRESUMO
Yasuní National Park in Ecuador is one of the most biodiverse places on earth. The fungi in this tropical rainforest are also diverse but have received little research attention. This research paper focuses on an important group of fungi in the family Polyporaceae and examines the genera Polyporus, Atroporus, and Neodictyopus that form aerial melanized cord-like structures called rhizomorphs. Phylogenetic analyses, macro and micromorphological descriptions of basidiomata and rhizomorphs, as well as cultural characterization were completed to better understand these ecologically important fungi. Here we describe four new species: Atroporus yasuniensis, Atroporus tagaeri, Neodictyopus sylvaticus, and Polyporus taromenane, and a new variety Polyporus leprieurii var. yasuniensis. The information presented in this study adds important new knowledge about the unusual rhizomorph producing fungi found in Yasuní National Park, Ecuador and other tropical rainforests.
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Ecossistema , Fungos/genética , Filogenia , Polyporaceae/genética , Animais , Biodiversidade , Equador , Fungos/classificação , Polyporaceae/classificação , Polyporus/genética , Simuliidae/genéticaRESUMO
Since mite allergens are the most relevant inducers of allergic diseases worldwide, resulting in significant morbidity and increased burden on health services, the International Collaboration in Asthma, Allergy and Immunology (iCAALL), formed by the American Academy of Allergy, Asthma and Immunology (AAAAI), the American College of Allergy, Asthma and Immunology (ACAAI), the European Academy of Allergy and Clinical Immunology (EAACI), and the World Allergy Organization (WAO), has proposed to issue an International Consensus (ICON) on the clinical consequences of mite hypersensitivity. The objectives of this document are to highlight aspects of mite biology that are clinically relevant, to update the current knowledge on mite allergens, routes of sensitization, the genetics of IgE responses to mites, the epidemiologic aspects of mite hypersensitivity, the clinical pictures induced by mites, the diagnosis, specific immunotherapeutic approaches, and prevention.
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BACKGROUND: Sublingual immunotherapy (SLIT) has become established in Europe, and its efficacy is being evaluated in the United States. The doses used for SLIT in Europe today are difficult to evaluate, because each manufacturer expresses the potency of its extracts differently. OBJECTIVES: To compare in vitro European SLIT maintenance solutions against US licensed standardized allergenic extract concentrates and to determine the monthly SLIT doses delivered expressed in bioequivalent allergy units ([B]AU). METHODS: We studied Dermatophagoides pteronyssinus, timothy grass pollen, cat (hair) and short ragweed pollen allergen extracts. The SLIT maintenance solutions of 4 leading European manufacturers and standardized concentrate extracts of 3 US manufacturers were analyzed with the following assays: protein content, relative potency (immunoglobulin E [IgE]-binding enzyme-linked immunosorbent assay [ELISA] inhibition) and major allergen content. The relative monthly allergen dose in (B)AU was calculated for each recommended SLIT schedule. RESULTS: Relative potency was approximately 10 times higher for US concentrate standardized extracts-which are meant to be diluted-than for European SLIT maintenance solutions of D pteronyssinus and timothy grass pollen. For cat (hair) and short ragweed pollen, the difference was less. Measurements of relative potency and major allergen content correlated well. In our assays, European mite extracts contain a very low quantity of Der p 2 compared with US mites. CONCLUSION: Recommended SLIT doses in Europe vary widely among the manufacturers, but are consistently lower (Eur1) or higher (Eur4) over all four allergens tested. SLIT efficacy probably depends on additional factors apart from the exact dose. SLIT dose finding studies should be done for each product.
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Alérgenos/farmacocinética , Dessensibilização Imunológica , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/imunologia , Preparações Farmacêuticas/administração & dosagem , Administração Sublingual , Alérgenos/administração & dosagem , Animais , Dessensibilização Imunológica/normas , Cálculos da Dosagem de Medicamento , Europa (Continente) , Humanos , Hipersensibilidade/epidemiologia , Indústrias/normas , Equivalência Terapêutica , Estados UnidosRESUMO
Osteocephalus festae is an Amazonian species recently resurrected from a synonymy with Osteocephalus buckleyi. Because few specimens are known, its morphological variation, diagnostic characters, and distribution are poorly understood. Herein we determine its phylogenetic relationships and provide a complete taxonomic account based on recently collected specimens (adults and larvae) from nine localities in Ecuador and Peru. Osteocephalus festae is most similar to Osteocephalus verruciger from which it differs in having less tuberculate dorsal skin on males, smaller tympanum, and more tooth rows in the oral disk of larvae. A phylogeny based on mitochondrial DNA sequences, genes 12S and ND1, shows that Osteocephalus festae is closely related to Osteocephalus buckleyi, Osteocephalus mutabor and Osteocephalus verruciger. A clade consisting of Osteocephalus festae, Osteocephalus verruciger, and Osteocephalus buckleyi is characterized by stream dwelling tadpoles. Surprisingly, we found paraphyly among Ecuadorian populations of Osteocephalus buckleyi and Osteocephalus verruciger. The causes for paraphyly are unknown but in Osteocephalus buckleyi may result from the existence of cryptic species.
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The secondary metabolite composition of the gorgonian octocoral Eunicea pinta from San Andrés Island, Colombia, is described for the first time. This investigation has resulted in the isolation of eight new gamma-cembranolide-type diterpenes, namely, compounds 3-10, and a new saponin, 13, possessing a pregnene-derived aglycon. In addition, two previously known alpha-methylene-gamma-lactone cembranolides, euniolide (1) and succinolide (2), were also isolated. Their structures were determined on the basis of the results of spectroscopic analysis, X-ray diffraction analysis, and chemical conversions. The discovery of cembranolides 3-10 has prompted us to conclude that the structures proposed earlier for several uprolide-type cembranolides need revision and that a minor stereochemical correction is also in order for succinolide. Cytotoxicity of compounds 3, 4, and 13 toward various cancer cell lines and results of an antimycobacterial screening for 2, 4, and 13 also are described.