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Classification of asteraceae weed pollen grains by image analysis
$90
Abstracts
7 Patients Patterns of Prick Skin Test Reactivity to Tree Pollen Allergens in with Seasonal Allergic Rhinitis Are Unrelated to Measured Pollen Exposure ,1. F. White l, D. I. Bernstein 2, M. Villareal 2, H. St. Clair 3, K. Murphy2; llmmunology, University of Cincinnati, Cincinnati, OH, 2University of Cincinnati, Cincinnati, OH, 3Department of Environmental Services, Cincinnati, OH. RATIONALE: The purpose of this study is to evaluate relationships between reported tree pollen exposure and in vivo percutaneous reactivities to commercially-available tree pollen extracts in the Cincinnati metropolitan area. METHODS: Sixty-seven patients were recruited who have resided in the Cincinnati area for most of their life with symptoms of seasonal allergic rhinitis during tree pollen season for at least two consecutive years. Patients underwent prick skin testing (PST) to'extracts of 15 regionally prevalent trees. Tree pollen counts from 1996-2002 were collected. Prevalences of reactivity to individual pollens and the estimated average wheal diameter were analyzed for correlations with tree pollen counts and hypersensitivity information to determine potential relationships. RESULTS: There was no significant correlation of mean pollen counts over 5 years with percent prevalence prick skin test reactivity to individual species or with mean wheal size diameter for individual tree pollen allergens. Kappa statistics were performed to examine agreement between skin test reactions to tree antigens base.d upon known patterns of crossreactivity. There were only modest levels of agreement between phylogenetically similar species such as box elder and red maple (~=0.37), and birch and oak (~=0.47). There was substantial agreement between white oak and red oak species 0r CONCLUSIONS: In this study, patterns of in vivo sensitization to pollen allergens are unrelated to ambient exposure to tree pollen allergen. These findings could be related to unknown cross-reactivity between tree pollen allergens or attributable to differences in relative allergenic potencies of individual pollens.
Funding: University of Cincinnati
8
Assessing Allergenicity of Indoor Air Fungal Contaminants
M. D. W. Ward l, M. E. Viana 2, N. Haykal-Coates l, L. B. Copeland l, S. H. Gavett I. M. K. SelgradO; 1Experimental Toxicology Division, US Environmental Protection Agency, Research Triangle Park, NC, 2College of Veterinary Medicine, North Carolina State University, Raleigh, NC. RATIONALE: The indoor environment has increased in importance to children's health with children now spending more than 90% of their time indoors. However, the health effects caused by childhood exposures to indoor molds are complex. M E T H O D S : We have assessed the allergic potential of two fungal extracts in a female BALB/c mouse model: (1) the biopesticide Metarhizium anisopliae (MACA) and (2) Stachybotrys chartarum (SCE-I). In a series of studies mice were subjected to 4 aspiration (ASP) exposures to 10 mg of MACA, SCE-I, bovine serum albumin (BSA) (negative control), or HBSS (vehicle control) over a 4-week period. Serum and bronchoalveolar lavage fluid (BALF) were collected before (DO), and at 1 (DI) and 3 (D3) days following the final ASP exposure. Additionally, wholebody plethysmography (Buxco) was used to assess pulmonary resistance and bronchoconstriction. Mice were assessed for immediate responses (10 minute baseline and 1 hour following each aspiration exposure) and airway hyperresponsiveness to methacholine (D1 and D3). RESULTS: Exposure to both extracts caused increased BALF total protein, LDH, and IL-5 levels (DI). BALF total cell counts as well as neutrophil (D 1), eosinophil and lymphocyte (D 1-3) counts were elevated. As was serum total IgE. Furthermore, these extracts caused significant increases in Buxco measures compared to both HBSS and BSA. Additionally, 4 IgE-inducing proteins have been identified in MACA. CONCLUSIONS: Respiratory exposure to either of these extracts causes responses similar to those observed in human allergic lung disease.
Funding: Supported by NCSU/EPA Cooperative Training Agreement CT826512010. This abstract does not reflect EPA policy.
J ALLERGY CLIN IMMUNOL FEBRUARY 2003
9 sisClassification of Asteraceae Weed Pollen Grains by Image AnalyT. C. Ong, X. Wang, B. W. Lee, H. T. W. Tan, F. T. Chew; National University of Singapore, Singapore, SINGAPORE. RATIONALE: Our work involves studying the feasibility of using image analysis to differentiate airborne pollen grains and spores. This study demonstrates the classification of Asteraceae weed pollen via this method. METHODS: A total of 20 types of Asteraceae weed pollen (Ambrosia sp., Artemisia sp. Baccharis sp. Chrysanthemum sp., Eupatorium sp., Helianthus sp., lva sp., Solidago sp., Taraxacum sp. and Xanthium sp.) were studied. Reference slides of respective Asteraceae weeds pollen were scanned using a CCD camera at 400• Image segmentation was performed prior to making measurements. 1000 individual pollen grains of each species were studied. A total of 47 primary and secondary parameters were measured but only 25 were used to classify the pollen grains by stepwise canonical discriminate analysis. RESULTS: 11 pollen types could be classified with accuracy > 9 0 % - -
Ambrosia acanthicarpa, Artemisia californica, Artemisia frigida, Artemisia vulgaris, Baccharis halimifolia, Baccharis sarothroides, Chrysanthemum leucanthemum, Helianthus annus, lva axillaries, Solidago spp. and Xanthium commune. 4 other pollen types namely Ambrosia artemisiifolia, Ambrosia psilostachya, Ambrosia trifida and lva xanthifolia fall into the above 80% accuracy range. Accuracy below 50% was obtained for 3 pollen types; Eupatorium capillifolium, Hymenoclea salsola and Taraxacum commune but could be improved with the use of texture-based parameters. CONCLUSIONS: Results of this study have demonstrated that different pollen types of the Asteraceae weed may be distinguished by image analysis. This method may form the basis for a rapid identification process of more airborne spores and pollen, and lead towards automating the airspora identification and quantification process.
Funding: Biomedical Research Council, Singapore
0
Identification of an Unknown Prevalent Fungal Airspora Component in Singapore via Internal Transcribed Spacer (ITS) and 18S rDNA Sequences
S. S. Joshi, T. C. Ong, A. H. B. Loo, F. L. Wong, T. K. Tan, F. T. Chew; National University of Singapore, Singapore, SINGAPORE. RATIONALE: In our previous studies, we isolated an unknown fungal spore that is prevalent in the Singapore environment. This study attempts to determine its identity and characterize its aerobiological profile and allergenicity. METHODS: Its aerobiological profile was monitored using a Burkard spore trap. The fungal spore was isolated and germinated in Sabouraud liquid medium but was resistant to sporulation even in several different media. A phylogenetic parsimony analysis using ITS (Internal Transcribed Spacer) and 18S rDNA sequences was thus carried out. Potential allergenicity of this spore was evaluated by skin prick testing. RESULTS: The unknown spore accounted for 21% and 17% of the outdoor and indoor airspora in Singapore, respectively (ranging from 18-972 spores/meter3/day). In the phylogenetic tree generated from ITS sequences of 35 fungal species, this unknown fungus was found in the clade consisting of Lophiostoma vagabundum (Lophiostomataceae) and Monodictys castaneae (Mitosporic Ascomycete). This clade was sister to members of Pleosporaceae. Likewise for the 18S tree consisting of 37 species, the unknown was found in a clade with members from Dothideomycetes (Pleosporales and Chaetothyriomycetes incertae sedis). Hence, the unknown fungus is thought to be a member of Dothideomycetes. A total of 10/58 atopic individuals responded with a minimum 3 • 3 mm skin prick reaction to extracts of this fungal spore compared to 0/10 among non-atopics (Fisher's, p<0.05). C O N C L U S I O N S : This prevalent fungi is likely a member of Dothideomycetes and is shown to be potentially allergenic. Further characterization would be required to understand its allergenicity in relation to other fungal spores.
Funding: Biomedical Research Council, Singapore
Abstracts
7 Patients Patterns of Prick Skin Test Reactivity to Tree Pollen Allergens in with Seasonal Allergic Rhinitis Are Unrelated to Measured Pollen Exposure ,1. F. White l, D. I. Bernstein 2, M. Villareal 2, H. St. Clair 3, K. Murphy2; llmmunology, University of Cincinnati, Cincinnati, OH, 2University of Cincinnati, Cincinnati, OH, 3Department of Environmental Services, Cincinnati, OH. RATIONALE: The purpose of this study is to evaluate relationships between reported tree pollen exposure and in vivo percutaneous reactivities to commercially-available tree pollen extracts in the Cincinnati metropolitan area. METHODS: Sixty-seven patients were recruited who have resided in the Cincinnati area for most of their life with symptoms of seasonal allergic rhinitis during tree pollen season for at least two consecutive years. Patients underwent prick skin testing (PST) to'extracts of 15 regionally prevalent trees. Tree pollen counts from 1996-2002 were collected. Prevalences of reactivity to individual pollens and the estimated average wheal diameter were analyzed for correlations with tree pollen counts and hypersensitivity information to determine potential relationships. RESULTS: There was no significant correlation of mean pollen counts over 5 years with percent prevalence prick skin test reactivity to individual species or with mean wheal size diameter for individual tree pollen allergens. Kappa statistics were performed to examine agreement between skin test reactions to tree antigens base.d upon known patterns of crossreactivity. There were only modest levels of agreement between phylogenetically similar species such as box elder and red maple (~=0.37), and birch and oak (~=0.47). There was substantial agreement between white oak and red oak species 0r CONCLUSIONS: In this study, patterns of in vivo sensitization to pollen allergens are unrelated to ambient exposure to tree pollen allergen. These findings could be related to unknown cross-reactivity between tree pollen allergens or attributable to differences in relative allergenic potencies of individual pollens.
Funding: University of Cincinnati
8
Assessing Allergenicity of Indoor Air Fungal Contaminants
M. D. W. Ward l, M. E. Viana 2, N. Haykal-Coates l, L. B. Copeland l, S. H. Gavett I. M. K. SelgradO; 1Experimental Toxicology Division, US Environmental Protection Agency, Research Triangle Park, NC, 2College of Veterinary Medicine, North Carolina State University, Raleigh, NC. RATIONALE: The indoor environment has increased in importance to children's health with children now spending more than 90% of their time indoors. However, the health effects caused by childhood exposures to indoor molds are complex. M E T H O D S : We have assessed the allergic potential of two fungal extracts in a female BALB/c mouse model: (1) the biopesticide Metarhizium anisopliae (MACA) and (2) Stachybotrys chartarum (SCE-I). In a series of studies mice were subjected to 4 aspiration (ASP) exposures to 10 mg of MACA, SCE-I, bovine serum albumin (BSA) (negative control), or HBSS (vehicle control) over a 4-week period. Serum and bronchoalveolar lavage fluid (BALF) were collected before (DO), and at 1 (DI) and 3 (D3) days following the final ASP exposure. Additionally, wholebody plethysmography (Buxco) was used to assess pulmonary resistance and bronchoconstriction. Mice were assessed for immediate responses (10 minute baseline and 1 hour following each aspiration exposure) and airway hyperresponsiveness to methacholine (D1 and D3). RESULTS: Exposure to both extracts caused increased BALF total protein, LDH, and IL-5 levels (DI). BALF total cell counts as well as neutrophil (D 1), eosinophil and lymphocyte (D 1-3) counts were elevated. As was serum total IgE. Furthermore, these extracts caused significant increases in Buxco measures compared to both HBSS and BSA. Additionally, 4 IgE-inducing proteins have been identified in MACA. CONCLUSIONS: Respiratory exposure to either of these extracts causes responses similar to those observed in human allergic lung disease.
Funding: Supported by NCSU/EPA Cooperative Training Agreement CT826512010. This abstract does not reflect EPA policy.
J ALLERGY CLIN IMMUNOL FEBRUARY 2003
9 sisClassification of Asteraceae Weed Pollen Grains by Image AnalyT. C. Ong, X. Wang, B. W. Lee, H. T. W. Tan, F. T. Chew; National University of Singapore, Singapore, SINGAPORE. RATIONALE: Our work involves studying the feasibility of using image analysis to differentiate airborne pollen grains and spores. This study demonstrates the classification of Asteraceae weed pollen via this method. METHODS: A total of 20 types of Asteraceae weed pollen (Ambrosia sp., Artemisia sp. Baccharis sp. Chrysanthemum sp., Eupatorium sp., Helianthus sp., lva sp., Solidago sp., Taraxacum sp. and Xanthium sp.) were studied. Reference slides of respective Asteraceae weeds pollen were scanned using a CCD camera at 400• Image segmentation was performed prior to making measurements. 1000 individual pollen grains of each species were studied. A total of 47 primary and secondary parameters were measured but only 25 were used to classify the pollen grains by stepwise canonical discriminate analysis. RESULTS: 11 pollen types could be classified with accuracy > 9 0 % - -
Ambrosia acanthicarpa, Artemisia californica, Artemisia frigida, Artemisia vulgaris, Baccharis halimifolia, Baccharis sarothroides, Chrysanthemum leucanthemum, Helianthus annus, lva axillaries, Solidago spp. and Xanthium commune. 4 other pollen types namely Ambrosia artemisiifolia, Ambrosia psilostachya, Ambrosia trifida and lva xanthifolia fall into the above 80% accuracy range. Accuracy below 50% was obtained for 3 pollen types; Eupatorium capillifolium, Hymenoclea salsola and Taraxacum commune but could be improved with the use of texture-based parameters. CONCLUSIONS: Results of this study have demonstrated that different pollen types of the Asteraceae weed may be distinguished by image analysis. This method may form the basis for a rapid identification process of more airborne spores and pollen, and lead towards automating the airspora identification and quantification process.
Funding: Biomedical Research Council, Singapore
0
Identification of an Unknown Prevalent Fungal Airspora Component in Singapore via Internal Transcribed Spacer (ITS) and 18S rDNA Sequences
S. S. Joshi, T. C. Ong, A. H. B. Loo, F. L. Wong, T. K. Tan, F. T. Chew; National University of Singapore, Singapore, SINGAPORE. RATIONALE: In our previous studies, we isolated an unknown fungal spore that is prevalent in the Singapore environment. This study attempts to determine its identity and characterize its aerobiological profile and allergenicity. METHODS: Its aerobiological profile was monitored using a Burkard spore trap. The fungal spore was isolated and germinated in Sabouraud liquid medium but was resistant to sporulation even in several different media. A phylogenetic parsimony analysis using ITS (Internal Transcribed Spacer) and 18S rDNA sequences was thus carried out. Potential allergenicity of this spore was evaluated by skin prick testing. RESULTS: The unknown spore accounted for 21% and 17% of the outdoor and indoor airspora in Singapore, respectively (ranging from 18-972 spores/meter3/day). In the phylogenetic tree generated from ITS sequences of 35 fungal species, this unknown fungus was found in the clade consisting of Lophiostoma vagabundum (Lophiostomataceae) and Monodictys castaneae (Mitosporic Ascomycete). This clade was sister to members of Pleosporaceae. Likewise for the 18S tree consisting of 37 species, the unknown was found in a clade with members from Dothideomycetes (Pleosporales and Chaetothyriomycetes incertae sedis). Hence, the unknown fungus is thought to be a member of Dothideomycetes. A total of 10/58 atopic individuals responded with a minimum 3 • 3 mm skin prick reaction to extracts of this fungal spore compared to 0/10 among non-atopics (Fisher's, p<0.05). C O N C L U S I O N S : This prevalent fungi is likely a member of Dothideomycetes and is shown to be potentially allergenic. Further characterization would be required to understand its allergenicity in relation to other fungal spores.
Funding: Biomedical Research Council, Singapore