title: Enzymatic activity in body and fecal extracts of the storage mite Chortoglyphus arcuatus.
abstract: BACKGROUND
  Chortoglyphus arcuatus has been described in many countries. Many allergens are potent enzymes, which may promote a Th2 immune response. The aim of this study was to evaluate the enzymatic activity of body and fecal extracts of C. arcuatus.
  
  
  METHODS
  Feces and bodies of full-grown C. arcuatus cultures were separated by sieving, extracted in PBS, dialyzed and lyophilized. The antigenic profile of both extracts was determined by SDS-PAGE. Immunoblot experiments were conducted using a pool of sera from allergic individuals residing in Galicia, a region of Spain, where this species is abundant. The enzymatic activity of the extracts was evaluated by the zymogram technique. Serine and cysteine protease activity was measured using in vitro methods. The API Zym system was used to determine the enzymatic properties of the extracts.
  
  
  RESULTS
  The antigenic profile showed that the body extract contained more and better defined bands than the fecal extract. Allergens were detected in both extracts in a molecular weight range between 14 and 100 kDa. Gelatinolytic gels confirmed that fecal extracts contain more hydrolytic enzymatic activity than body extracts. Serine protease activity in fecal extracts was higher than in body extracts (5.98 vs. 2.701 IU of trypsin/mg of freeze-dried material). No cysteine protease activity was detected.
  
  
  CONCLUSIONS
  C. arcuatus extracts contain several allergens and proteins with high enzymatic activity, especially in the feces. Some of these allergens may be enzymes. Fecal extracts have more enzymatic activity than body extracts.
authors.affiliation: Hospital Xeral de Calde, Allergology Service, Lugo, Spain. jcarnes@leti.com
authors.forename: Jerónimo
authors.initials: J
authors.lastname: Carnés
authors.forename: Manuel
authors.initials: M
authors.lastname: Boquete
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Carballada
authors.forename: Victor
authors.initials: V
authors.lastname: Iraola
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Gallego
authors.forename: Enrique
authors.initials: E
authors.lastname: Fernández-Caldas
mesh_terms: D000485:Allergens / Q000032:analysis / Q000737:chemistry; D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism; D005243:Feces / Q000201:enzymology*; D006798:Housing; D006801:Humans; D015151:Immunoblotting; D008925:Mites / Q000201:enzymology* / Q000276:immunology; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000378:metabolism; D013030:Spain; D014020:Tissue Extracts / Q000378:metabolism
chemical_list: D000485:Allergens; D014020:Tissue Extracts; D012697:Serine Endopeptidases; D003546:Cysteine Endopeptidases