title: Enzymatic isolation of Lacazia loboi cells from skin lesions of lobomycosis.
abstract: Lacazia loboi is the etiologic agent of Jorge Lobo's disease, a cutaneous and subcutaneous mycosis endemic to Latin America tropical regions and characterized by chronic nodular or keloidal lesions which develop after traumatic events. A new method for the extraction of L. loboi yeast cells from biopsies of lobomycosis skin lesions is presented. The method is based on the proteolytic action of the enzyme dispase which is known for its action against fibronectin and collagen type IV. Fungal identification was based on histological examination of the biological material and molecular analysis based on 18S ribosomal sequences. Observations under optic and fluorescence microscopy proved the efficacy of enzymatic isolation of the lobomycosis etiologic agent, as well as identifying the organism's main parasitic characteristics. Molecular phylogenetic analysis corroborated the histological examination and indicated L. loboi relationship with other members of the Onygenales. Use of dispase proved to be ideal for the isolation of L. loboi from human biopsies, shows promise as an important tool for improving biological studies of this peculiar fungus.
authors.affiliation: Laboratório de Dermato-Imunologia, Universidade do Estado do Pará, Universidade Federal do Pará and Centro de Referência e Treinamento em Dermatologia Sanitária do Estado do Pará "Dr. Marcello Candia" (MC), Marituba, Pará, Brazil. csalgado@ufpa.br
authors.forename: Claudio Guedes
authors.initials: CG
authors.lastname: Salgado
authors.forename: Lívia Simone Alves
authors.initials: LS
authors.lastname: Tavares
authors.forename: Helio Longoni
authors.initials: HL
authors.lastname: Plautz
authors.forename: Moises Batista
authors.initials: MB
authors.lastname: Da Silva
authors.forename: Suellen Sirleide Pereira
authors.initials: SS
authors.lastname: Yamano
authors.forename: Patricia Fagundes
authors.initials: PF
authors.lastname: Da Costa
authors.forename: Igor Guerreiro
authors.initials: IG
authors.lastname: Hamoy
authors.forename: Anderson Nonato Do Rosario
authors.initials: AN
authors.lastname: Marinho
authors.forename: Andrea Kely Campos Ribeiro Dos
authors.initials: AK
authors.lastname: Santos
authors.forename: Ubirajara Imbiriba
authors.initials: UI
authors.lastname: Salgado
authors.forename: Jorge Pereira
authors.initials: JP
authors.lastname: Da Silva
mesh_terms: D001706:Biopsy; D003881:Dermatomycoses / Q000175:diagnosis / Q000382:microbiology*; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009172:Mycology / Q000379:methods; D020036:Onygenales / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012867:Skin / Q000378:metabolism* / Q000382:microbiology* / Q000473:pathology
chemical_list: D012337:RNA, Ribosomal, 18S; D010450:Endopeptidases; C023139:dispase