6 results on '"Garces, Hans Garcia"'
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2. Paracoccidioides brasiliensis Isolated from Nine-Banded Armadillos (Dasypus novemcinctus) Reveal Population Structure and Admixture in the Amazon Basin
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Bagagli, Eduardo, primary, Matute, Daniel Ricardo, additional, Garces, Hans Garcia, additional, Tenório, Bernardo Guerra, additional, Garces, Adalberto Garcia, additional, Alves, Lucas Gomes de Brito, additional, Yamauchi, Danielle Hamae, additional, Hrycyk, Marluce Francisca, additional, Barker, Bridget Marie, additional, and Teixeira, Marcus de Melo, additional
- Published
- 2021
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3. Prp8 intein in dermatophytes: molecular identification and evolution
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Garces, Hans Garcia, Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
- Subjects
filogenia ,Dermatophytes ,Intein PRP8 ,molecular identification ,PRP8 intein ,Dermatófitos ,Identificação molecular ,phylogeny - Abstract
Submitted by HANS GARCIA GARCES null (atiweb@gmail.com) on 2018-02-15T14:17:44Z No. of bitstreams: 1 tese final hans garcia.pdf: 5207018 bytes, checksum: dc8235a547fe1b9abb0de219b802a19f (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-02-20T17:09:18Z (GMT) No. of bitstreams: 1 garces_hg_dr_bot_par.pdf: 414383 bytes, checksum: a87b367ff5d75b8f43af0ee14876ad21 (MD5) Made available in DSpace on 2018-02-20T17:09:18Z (GMT). No. of bitstreams: 1 garces_hg_dr_bot_par.pdf: 414383 bytes, checksum: a87b367ff5d75b8f43af0ee14876ad21 (MD5) Previous issue date: 2018-02-08 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Os dermatófitos são um grupo de fungos constituídos pelos gêneros Trichophyton, Epidermophyton e Microsporum que têm a habilidade de degradar a queratina. É por essa razão que podem colonizar a pele do homem e dos animais, embora também possam crescer no ambiente, geralmente em solos com restos de queratina. As características morfológicas permitem a identificação e classificação taxonômica das espécies, mas realizar um diagnóstico baseado nestas características pode levar a erros. As técnicas moleculares ajudam a realizar diagnósticos mais rápidos baseados em uma identificação molecular e também têm possibilitado importantes avanços quanto aos estudos filogenéticos, sendo as sequências ITS1-5.8S-ITS2 as mais utilizadas. No entanto, estes marcadores não são suficientes para identificar e elucidar todas as relações filogenéticas e taxonômicas dos dermatófitos devido à ampla variedade de espécies existentes, -sendo necessário novos marcadores genéticos, como o intein PRP8. Os inteins são elementos genéticos parasitas, de natureza proteica, presentes em alguns fungos e estão associados a importantes genes altamente conservados. Ointein PRP8 está localizado nogenePRP8 que codifica para a proteína PRP8 associada ao complexo do spliciosoma. O presente estudo visa caracterizar o intein PRP8 em fungos dermatófitos, de forma a empregar estes elementos genéticos como marcadores moleculares para uma correta identificação destas espécies e elucidações das relações filogenéticas do grupo. Para realizar este objetivo, foram caracterizadasmolecularmente 45 cepas usando as regiões nucleares ribossomais ITS1-5.8S-ITS2 e D1/D2 e posteriormente a região do intein PRP8 de 40 cepas foi sequenciada para determinar as características do intein. A presença do intein foi comprovada em todas as espécies avaliadas, apresentando um full-intein com uma Homing Endonuclease presumivelmente ativa. As construções filogenéticas obtidas usando as sequências do intein são correspondentes as construídas a partir de genes nucleares ribossomais, demonstrando a validade do intein para ser incluído em futuros estudos filogenético associados a taxonomia e determinação de espécies. O polimorfismo na região da Homing Endonuclease permitiu a identificação de algumas espécies de Microsporum mediante um ensaio de PCR-Eletroforese. Dermatophytes are a fungal group composed by the genera Trichophyton, Epidermophyton and Microsporum with the ability to degrade keratin. That is why they can colonize the skin of man and animals, although they can also grow in the environment, usually in soils with remains of keratin. The morphological characteristics allow the identification and taxonomic classification of the species, but a diagnosis based on these characteristics is difficult. Molecular techniques made diagnoses based on molecular identification faster and made possible important advances in phylogenetic studies, with ITS1-5.8s-ITS2 sequences being the most used. However, these markers are not enough to elucidate all the phylogenetic relationships and taxonomic of dermatophytes due to the wide variety of existing species, and new genetic markers may be needed for correct identification and phylogenetic studies, such as the PRP8 intein. Inteins are parasitic genetic elements of a protein nature present in some fungi and are associated with important highly conserved genes. The PRP8 intein is located within the PRP8 gene coding for the PRP8 protein associated with the spliciosome complex. The present study aims to characterize the PRP8 intein in dermatophyte fungi, in order to use these genetic elements as molecular markers for a correct identification of these species and elucidations of the phylogenetic relationships of the group. To accomplish this goal, 45 strains were molecularly characterized using the ITS1-5.8S-ITS2 and D1 / D2 ribosomal nuclear regions and subsequently the PRP8 intein region of 40 strains was sequenced to determine the intein characteristics. The presence of intein was confirmed in all evaluated species, presenting a full-intein with a presumably active Homing Endonuclease. The phylogenetic construction obtained using the intein sequences correspond to those constructed from ribosomal nuclear genes, demonstrating the intein validity to be included in future phylogenetic studies associated with taxonomy and species determination. Polymorphism in the Homing Endonuclease region allowed the identification of some Microsporum species by a PCR-Electrophoresis assay. 12481-13-0
- Published
- 2018
4. Chemical Composition and Bioactivity of Essential Oil from Blepharocalyx salicifolius.
- Author
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Furtado, Fabiana Barcelos, Borges, Bruna Cristina, Teixeira, Thaise Lara, Garces, Hans Garcia, Almeida Junior, Luiz Domingues de, Alves, Fernanda Cristina Bérgamo, Silva, Claudio Vieira da, and Fernandes Junior, Ary
- Subjects
ESSENTIAL oils ,AROMATIC plants ,EVERGREENS ,ANTIFUNGAL agents ,ANTIBACTERIAL agents ,CHEMICAL composition of plants - Abstract
Natural products represent a source of biologically active molecules that have an important role in drug discovery. The aromatic plant Blepharocalyx salicifolius has a diverse chemical constitution but the biological activities of its essential oils have not been thoroughly investigated. The aims of this paper were to evaluate in vitro cytotoxic, antifungal and antibacterial activities of an essential oil from leaves of B. salicifolius and to identify its main chemical constituents. The essential oil was extracted by steam distillation, chemical composition was determined by gas chromatography/mass spectrometry, and biological activities were performed by a microdilution broth method. The yield of essential oil was 0.86% (w/w), and the main constituents identified were bicyclogermacrene (17.50%), globulol (14.13%), viridiflorol (8.83%), γ-eudesmol (7.89%) and α-eudesmol (6.88%). The essential oil was cytotoxic against the MDA-MB-231 (46.60 μg·mL
-1 ) breast cancer cell line, being more selective for this cell type compared to the normal breast cell line MCF-10A (314.44 μg·mL-1 ). Flow cytometry and cytotoxicity results showed that this oil does not act by inducing cell death, but rather by impairment of cellular metabolism specifically of the cancer cells. Furthermore, it presented antifungal activity against Paracoccidioides brasiliensis (156.25 μg·mL-1 ) but was inactive against other fungi and bacteria. Essential oil from B. salicifolius showed promising biological activities and is therefore a source of molecules to be exploited in medicine or by the pharmaceutical industry. [ABSTRACT FROM AUTHOR]- Published
- 2018
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5. Evolution of species of the genus sporothrix based on the sequences of vma and prp8 inteins
- Author
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Oliveira, Alana Lucena, Universidade Estadual Paulista (Unesp), Bosco, Sandra de Moraes Gimenes [UNESP], and Garces, Hans Garcia
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Sporotrix spp ,Filogenia ,Phylogen ,Intein VMA ,Diagnóstico molecular ,Intein PRP8 ,Molecular diagnosis - Abstract
Submitted by Alana Lucena Oliveira (alana.lucena@unesp.br) on 2021-02-01T15:48:01Z No. of bitstreams: 1 Dissertação Alana Lucena Oliveira 2020.pdf: 2308331 bytes, checksum: bd160b6361208756396acd7a6fa7d08b (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2021-02-02T18:55:17Z (GMT) No. of bitstreams: 1 oliveira_al_me_bot_par.pdf: 2455167 bytes, checksum: f38020f6125c00265711f879b2caa596 (MD5) Made available in DSpace on 2021-02-02T18:55:17Z (GMT). No. of bitstreams: 1 oliveira_al_me_bot_par.pdf: 2455167 bytes, checksum: f38020f6125c00265711f879b2caa596 (MD5) Previous issue date: 2020-10-30 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) A esporotricose é uma micose causada principalmente pelas espécies do clado patogênico do gênero Sporothrix. Estas espécies compreendem S. schenckii, S. brasiliensis, S. globosa e S. luriei. S. schenckii tem uma ampla distribuição mundial enquanto S. globosa é uma sapronose comum na Ásia e S. luriei é uma espécie bem restrita e com pouquíssimos relatos. A esporotricose causada por S. brasiliensis é uma zoonose que afeta principalmente os gatos, cuja incidência aumentou no Brasil na última década, sendo considerada uma das micoses emergentes mais importantes na atualidade. Visando a eliminação de surtos, junto a campanhas de prevenção e medidas de vigilância, é muito importante contar com métodos diagnósticos de baixo custo, rápidos e precisos compreendendo as corretas espécies envolvidas. Foi realizada a padronização e diferenciação das espécies do clado patogênico do gênero Sporothrix por meio de biologia molecular utilizando regiões gênicas específicas dos inteins VMA e PRP8 que ainda não foram estudas para essas espécies. A região dos inteins VMA e PRP8 já foram utilizadas com finalidades de diagnóstico e estudos filogenéticos em outros fungos, sendo, portanto, uma região promissora para esses estudos no gênero Sporothrix Inteins são sequencias de proteínas presentes em alguns organismos que têm a capacidade de realizar um processo de auto-splicing logo após a tradução da sua proteína hospedeira. Sendo comprovada a existência dos dois inteins (VMA e PRP8) na maioria das espécies do clado patogênico de Sporothrix spp., o objetivo foi caracterizar os inteins PRP8 e VMA visando inferências diagnósticas e evolutivas, incluindo amostras de DNA a partir de isolados em cultivo . Até então, analisamos 22 cepas de isolados clínicos (5 S. globosa, 5 S. schenckii, 12 S. brasiliensis) pelo sequenciamento das regiões dos inteins com primers por nós desenhados e analisamos por bioinformática uma cepa de S. luriei. O intein PRP8 foi encontrado em todas as espécies enquanto o VMA foi ausente em S. globosa e S. luriei. Os inteins sempre foram encontrados na forma de Full-length intein. As análises filogenéticas permitiram a diferenciação das espécies de S. schenckii próximo a S. brasiliensis e S. globosa mais próxima de S. luriei ao analisar as sequencias do intein PRP8. Igualmente, o intein VMA evidenciou um agrupamento separado das espécies S. schenckii e S. brasiliensis. A presença ou ausência e análise do polimorfismo encontrado no intein VMA permitiu desenhar um par de primers adicional para diferenciar as espécies S. schenckii, S. brasiliensis e S. globosa por meio de PCR/eletroforese, sem necessidade de sequenciamento de DNA. Sporotrichosis is a fungal disease caused mainly by species of the pathogenic clade of the genus Sporothrix. These species comprise S. schenckii, S. brasiliensis, S. globosa and S. luriei. S. schenckii has a broad worldwide distribution while S. globosa is a common sapronosis in Asia and S. luriei is a very restricted species with very few reports associated with disease. S. brasiliensis is a zoonosis that mainly affects cats and has had a great incidence in Brazil in the last decade, being considered one of the most important emerging mycoses in recent times. In order to eliminate outbreaks, along with prevention campaigns and surveillance measures, it is very important to have low-cost, fast and accurate diagnostic methods, identifying the correct species involved. Standardization and differentiation of the species from the pathogenic clade of the complex genus Sporothrix were carried out by means of molecular biology using specific gene regions of the VMA and PRP8 inteins that have not yet been studied within these species. The VMA and PRP8 intein regions have already been used for the purposes of diagnosis and phylogenetic studies in other fungi, and are thus considered promising regions for the genus Sporothrix. Inteins are protein sequences present in some organisms that have the ability to perform a self-splicing process just after their host protein is translated. Since the existence of both inteins is proven in the majority of all species of the Sporothrix pathogenic clade, we aim to characterize the PRP8 and VMA inteins in order to diagnose these species molecularly and to determine the evolutionary history of the group, including DNA samples from cultivated isolates. Currently, we have 22 strains of clinical isolates (5 S. globosa, 5 S. schenckii, 12 S. brasiliensis) by sequencing the regions with designed primers and one strain of S. luriei that was analyzed by bioinformatics. The PRP8 intein was found in all species while the VMA intein was absent in S. globosa and S. luriei. Inteins was always found in full-length form. Phylogenetic analyses allowed species differentiation by grouping S. schenckii close to S. brasiliensis and S. globosa close to S. luriei when analyzing the sequences of the PRP8 intein. Likewise, the VMA intein showed a separate clustering of the species S. schenckii and S. brasiliensis. The absence of inteins in S. globosa and the polymorphism found in the VMA intein allowed the designing of an additional primer set to differentiate the species S. schenkii, S. brasiliensis and S. globosa thought only a PCR/Eletrophoresis technique without the need for sequencing. FAPESP: 2019/3489-7
- Published
- 2020
6. Antifungal activity of liriodenine on agents of systemic mycoses, with emphasis on the genus Paracoccidioides .
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Vinche ADL, de-la-Cruz-Chacón I, González-Esquinca AR, da Silva JF, Ferreira G, Dos Santos DC, Garces HG, de Oliveira DVM, Marçon C, Cavalcante RS, and Mendes RP
- Abstract
Background: Endemic systemic mycoses remain a health challenge, since these opportunistic diseases are increasingly infecting immunosuppressed patients. The simultaneous use of antifungal compounds and other drugs to treat infectious or non-infectious diseases has led to several interactions and undesirable effects. Thus, new antifungal compounds should be investigated. The present study aimed to evaluate the activity of liriodenine extracted from Annona macroprophyllata on agents of systemic mycoses, with emphasis on the genus Paracoccidioides ., Methods: The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined by the microdilution method. The cellular alterations caused by liriodenine on a standard P. brasiliensis (Pb18) strain were evaluated by transmission and scanning electron microscopy., Results: Liriodenine was effective only in 3 of the 8 strains of the genus Paracoccidioides and in the Histoplasma capsulatum strain, in a very low concentration (MIC of 1.95 μg.mL
-1 ); on yeasts of Candida spp. (MIC of 125 to 250 μg.mL-1), including C. krusei (250 μg.mL-1 ), which has intrinsic resistance to fluconazole; and in Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 μg.mL-1 ). However, liriodenine was not effective against Aspergillus fumigatus at the studied concentrations. Liriodenine exhibited fungicidal activity against all standard strains and clinical isolates that showed to be susceptible by in vitro tests. Electron microscopy revealed cytoplasmic alterations and damage to the cell wall of P. brasiliensis (Pb18)., Conclusion: Our results indicate that liriodenine is a promising fungicidal compound that should undergo further investigation with some chemical modifications., Competing Interests: Competing interests: The authors declare that they have no competing interests.- Published
- 2020
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