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1.
The Korean Journal of Parasitology ; : 85-93, 2015.
Article in English | WPRIM | ID: wpr-130560

ABSTRACT

Proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of proteome. For detection of antigens from Haemaphysalis longicornis, 1-dimensional electrophoresis (1-DE) quantitative immunoblotting technique combined with 2-dimensional electrophoresis (2-DE) immunoblotting was used for whole body proteins from unfed and partially fed female ticks. Reactivity bands and 2-DE immunoblotting were performed following 2-DE electrophoresis to identify protein spots. The proteome of the partially fed female had a larger number of lower molecular weight proteins than that of the unfed female tick. The total number of detected spots was 818 for unfed and 670 for partially fed female ticks. The 2-DE immunoblotting identified 10 antigenic spots from unfed females and 8 antigenic spots from partially fed females. Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF) of relevant spots identified calreticulin, putative secreted WC salivary protein, and a conserved hypothetical protein from the National Center for Biotechnology Information and Swiss Prot protein sequence databases. These findings indicate that most of the whole body components of these ticks are non-immunogenic. The data reported here will provide guidance in the identification of antigenic proteins to prevent infestation and diseases transmitted by H. longicornis.


Subject(s)
Animals , Antigens/analysis , Arthropod Proteins/analysis , Electrophoresis , Immunoblotting , Ixodidae/chemistry , Mass Screening , Proteomics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
2.
The Korean Journal of Parasitology ; : 85-93, 2015.
Article in English | WPRIM | ID: wpr-130553

ABSTRACT

Proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of proteome. For detection of antigens from Haemaphysalis longicornis, 1-dimensional electrophoresis (1-DE) quantitative immunoblotting technique combined with 2-dimensional electrophoresis (2-DE) immunoblotting was used for whole body proteins from unfed and partially fed female ticks. Reactivity bands and 2-DE immunoblotting were performed following 2-DE electrophoresis to identify protein spots. The proteome of the partially fed female had a larger number of lower molecular weight proteins than that of the unfed female tick. The total number of detected spots was 818 for unfed and 670 for partially fed female ticks. The 2-DE immunoblotting identified 10 antigenic spots from unfed females and 8 antigenic spots from partially fed females. Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF) of relevant spots identified calreticulin, putative secreted WC salivary protein, and a conserved hypothetical protein from the National Center for Biotechnology Information and Swiss Prot protein sequence databases. These findings indicate that most of the whole body components of these ticks are non-immunogenic. The data reported here will provide guidance in the identification of antigenic proteins to prevent infestation and diseases transmitted by H. longicornis.


Subject(s)
Animals , Antigens/analysis , Arthropod Proteins/analysis , Electrophoresis , Immunoblotting , Ixodidae/chemistry , Mass Screening , Proteomics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
3.
Clinics ; 66(7): 1199-1202, 2011. ilus
Article in English | LILACS | ID: lil-596908

ABSTRACT

INTRODUCTION: Asthma affects approximately 10 percent of the world's population. Sensitization to allergens is an important risk factor, and exposure to allergens is associated with disease severity. METHODS: We performed skin tests to evaluate allergen sensitization to mites, cockroaches, cats, dogs, and molds in 73 asthmatic patients. Enzyme Linked Immunosorbent Assay was used to assay the mite and cockroach allergens found in dust from the bedding, hammocks, bedroom floors, living rooms, and kitchens of 29 patients and 14 controls. RESULTS: Fifty patients (68.5 percent) had positive skin test responses. There were positive responses to D. pteronyssinus (52.0 percent), B. tropicalis (53.4 percent), T. putrescentiae (15.0 percent), E. maynei (12.3 percent), L. destructor (8.2 percent), B. germanica (20.5 percent), P. americana (21.9 percent), Felis catus (10.9 percent), C. herbarium (2.7 percent), A. alternata (4.1 percent), and P. notatun (1.3 percent). The exposure to mite and cockroach allergens was similar in the patients and the controls. The Dermatophagoides pteronyssinus Group 1 levels were highest in the beds and hammocks. The Blattella germanica Group 1 levels were highest in the kitchens, living rooms and hammocks. DISCUSSION: The positive skin tests to mites, cockroaches and cats were consistent with previous studies. D pteronyssinus was the most prevalent home dust mite, and hammocks were a source of allergens. To improve asthma prophylaxis, it is important to determine its association with mite allergen exposure in hammocks.


Subject(s)
Adolescent , Adult , Animals , Female , Humans , Male , Middle Aged , Young Adult , Allergens/immunology , Asthma/immunology , Dust/immunology , Antigens, Dermatophagoides/analysis , Arthropod Proteins/analysis , Case-Control Studies , Cockroaches , Cysteine Endopeptidases/analysis , Dust/analysis , Enzyme-Linked Immunosorbent Assay , Mites , Risk Factors , Skin Tests , Statistics, Nonparametric
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