Progress 10/01/12 to 09/30/13
Outputs
Progress Report Objectives (from AD-416): The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Initial research conducted at The MITRE Corporation (MITRE) was patented and licensed to Quickpath Bioscience. Approach (from AD-416): Fimbriae, hair-like structures produced by many species of bacteria, vary in the composition of the fimbrial shaft and protein adhesins found on the tips. Bacterial adhesins selectively bind to tissue-specific glycans. For example, pathogens such as Salmonella and E. coli expressing Type I fimbriae attach to tissues/micelles coated with mannose. There are about twenty-two known fimbrial types. However, reproducibility and predictability of attachment is uncertain. Little is known about how the environment influences fimbrial production. Furthermore, Type I fimbriae are distributed among many bacterial species and there can be variations in the shaft fimbrin protein while the tip adhesin protein remains the same. We have selected a model strain, uropathogenic Escherichia coli (UPEC), to which we will obtain monoclonal antibodies produced against the target fimbriae. Immunoassay studies elucidating fimbrial expression as a function of environmental parameters will be conducted with methods developed for the Signalyte spectrofluorimeter(Creativ MicroTech) and imaged by Epifluorescent and Atomic Force Microscopy (NIST). Environmental parameters to be investigated include: pH (including shifts in pH), redox, temperature, culture age, nutrient ratios (carbon:nitrogen: phosphorus), nutrient composition, ionic strength, and cell chemical signaling effects. After establishing environmental conditions that reliably produce fimbrial expression, attachment studies to lipid-filled glycoprotein micelles provided by MITRE will be performed to establish optimal binding kinetics and survival. Target fimbriae will also be screened against available glycan microarrays. The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices. Fimbriae play a crucial role in attachment of bacteria to biotic (plant) and abiotic surfaces and formation of biofilms. An understanding of these processes is critical to understanding survival and to developing intervention strategies to prevent contamination of fresh produce. The role of Mitre is to provide glycoprotein- or glycolipid-coated micelles for experimentation and to provide expertise on adhesin-receptor binding. Studies confirm that glycoprotein micelles can be used to capture pathogenic E. coli and Salmonella. However, implementation of fimbrial-based capture/detection technologies will require additional information on environmental determinant of fimbrial expression.
Impacts
(N/A)
Publications
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Initial research conducted at The MITRE Corporation (MITRE) was patented and licensed to Quickpath Bioscience. Approach (from AD-416): Fimbriae, hair-like structures produced by many species of bacteria, vary in the composition of the fimbrial shaft and protein adhesins found on the tips. Bacterial adhesins selectively bind to tissue-specific glycans. For example, pathogens such as Salmonella and E. coli expressing Type I fimbriae attach to tissues/micelles coated with mannose. There are about twenty-two known fimbrial types. However, reproducibility and predictability of attachment is uncertain. Little is known about how the environment influences fimbrial production. Furthermore, Type I fimbriae are distributed among many bacterial species and there can be variations in the shaft fimbrin protein while the tip adhesin protein remains the same. We have selected a model strain, uropathogenic Escherichia coli (UPEC), to which we will obtain monoclonal antibodies produced against the target fimbriae. Immunoassay studies elucidating fimbrial expression as a function of environmental parameters will be conducted with methods developed for the Signalyte spectrofluorimeter(Creativ MicroTech) and imaged by Epifluorescent and Atomic Force Microscopy (NIST). Environmental parameters to be investigated include: pH (including shifts in pH), redox, temperature, culture age, nutrient ratios (carbon:nitrogen: phosphorus), nutrient composition, ionic strength, and cell chemical signaling effects. After establishing environmental conditions that reliably produce fimbrial expression, attachment studies to lipid-filled glycoprotein micelles provided by MITRE will be performed to establish optimal binding kinetics and survival. Target fimbriae will also be screened against available glycan microarrays. The role of Mitre is to provide glycoprotein- or glycolipid-coated micelles for experimentation and to provide expertise on adhesin-receptor binding. Fimbriae play a crucial role in attachment of bacteria to biotic (plant) and abiotic surfaces and formation of biofilms. An understanding of these processes is critical to understanding survival and to developing intervention strategies to prevent contamination of fresh produce.
Impacts
(N/A)
Publications
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Initial research conducted at The MITRE Corporation (MITRE) was patented and licensed to Quickpath Bioscience. Approach (from AD-416) Fimbriae, hair-like structures produced by many species of bacteria, vary in the composition of the fimbrial shaft and protein adhesins found on the tips. Bacterial adhesins selectively bind to tissue-specific glycans. For example, pathogens such as Salmonella and E. coli expressing Type I fimbriae attach to tissues/micelles coated with mannose. There are about twenty-two known fimbrial types. However, reproducibility and predictability of attachment is uncertain. Little is known about how the environment influences fimbrial production. Furthermore, Type I fimbriae are distributed among many bacterial species and there can be variations in the shaft fimbrin protein while the tip adhesin protein remains the same. We have selected a model strain, uropathogenic Escherichia coli (UPEC), to which we will obtain monoclonal antibodies produced against the target fimbriae. Immunoassay studies elucidating fimbrial expression as a function of environmental parameters will be conducted with methods developed for the Signalyte spectrofluorimeter(Creativ MicroTech) and imaged by Epifluorescent and Atomic Force Microscopy (NIST). Environmental parameters to be investigated include: pH (including shifts in pH), redox, temperature, culture age, nutrient ratios (carbon:nitrogen: phosphorus), nutrient composition, ionic strength, and cell chemical signaling effects. After establishing environmental conditions that reliably produce fimbrial expression, attachment studies to lipid-filled glycoprotein micelles provided by MITRE will be performed to establish optimal binding kinetics and survival. Target fimbriae will also be screened against available glycan microarrays. Virtually all bacteria possess one or more fimbriae or pili, hair-like structures extending from the cell surface. These fimbriae/pili allow for attachment to various surfaces, including a variety of inert surfaces. Fimbriae/pili associated with pathogens possess adhesins that are responsible for specific attachment to receptor molecules found on human tissues; these are typically glycoproteins or glycol lipids. The role of Mitre is to provide glycoprotein- or glycolipid-coated micelles for experimentation and to provide expertise on adhesin-receptor binding. Meetings are conducted monthly.
Impacts
(N/A)
Publications
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Initial research conducted at The MITRE Corporation (MITRE) was patented and licensed to Quickpath Bioscience. Approach (from AD-416) Fimbriae, hair-like structures produced by many species of bacteria, vary in the composition of the fimbrial shaft and protein adhesins found on the tips. Bacterial adhesins selectively bind to tissue-specific glycans. For example, pathogens such as Salmonella and E. coli expressing Type I fimbriae attach to tissues/micelles coated with mannose. There are about twenty-two known fimbrial types. However, reproducibility and predictably of attachment is uncertain. Little is known about how the environment influences fimbrial production. Furthermore, Type I fimbriae are distributed among many bacterial species and there can be variations in the shaft fimbrin protein while the tip adhesin protein remains the same. We have selected a model strain, uropathogenic Escherichia coli (UPEC), to which we will obtain monoclonal antibodies produced against the target fimbriae. Immunoassay studies elucidating fimbrial expression as a function of environmental parameters will be conducted with methods developed for the Signalyte spectrofluorimeter (Creativ MicroTech) and imaged by Epifluorescent and Atomic Force Microscopy (NIST). Environmental parameters to be investigated include: pH (including shifts in pH), redox, temperature, culture age, nutrient ratios (carbon:nitrogen: phosphorus), nutrient composition, ionic strength, and cell chemical signaling effects. After establishing environmental conditions that reliably produce fimbrial expression, attachment studies to lipid-filled glycoprotein micelles provided by MITRE will be performed to establish optimal binding kinetics and survival. Target fimbriae will also be screened against available glycan microarrays. The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Virtually all bacteria possess one or more fimbriae or pili, hair-like structures extending from the cell surface. These fimbriae/pili allow for attachment to various surfaces, including a variety of inert surfaces. Fimbriae/pili associated with pathogens possess adhesions that are responsible for specific attachment to receptor molecules found on human tissues, typically at the site of infection. For example, uropathogenic E. coli (UPEC) possess Type I fimbriae that mediate attachment to mannose-containing glycoproteins coating the urinary tract. The role of Mitre is to provide glycoprotein- coated micelles for experimentation and to provide expertise on adhesion- receptor binding. Meetings are conducted monthly.
Impacts
(N/A)
Publications
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Initial research conducted at The MITRE Corporation (MITRE) was patented and licensed to Quickpath Bioscience. Approach (from AD-416) Fimbriae, hair-like structures produced by many species of bacteria, vary in the composition of the fimbrial shaft and protein adhesins found on the tips. Bacterial adhesins selectively bind to tissue-specific glycans. For example, pathogens such as Salmonella and E. coli expressing Type I fimbriae attach to tissues/micelles coated with mannose. There are about twenty-two known fimbrial types. However, reproducibility and predictably of attachment is uncertain. Little is known about how the environment influences fimbrial production. Furthermore, Type I fimbriae are distributed among many bacterial species and there can be variations in the shaft fimbrin protein while the tip adhesin protein remains the same. We have selected a model strain, uropathogenic Escherichia coli (UPEC), to which we will obtain monoclonal antibodies produced against the target fimbriae. Immunoassay studies elucidating fimbrial expression as a function of environmental parameters will be conducted with methods developed for the Signalyte spectrofluorimeter (Creativ MicroTech) and imaged by Epifluorescent and Atomic Force Microscopy (NIST). Environmental parameters to be investigated include: pH (including shifts in pH), redox, temperature, culture age, nutrient ratios (carbon:nitrogen: phosphorus), nutrient composition, ionic strength, and cell chemical signaling effects. After establishing environmental conditions that reliably produce fimbrial expression, attachment studies to lipid-filled glycoprotein micelles provided by MITRE will be performed to establish optimal binding kinetics and survival. Target fimbriae will also be screened against available glycan microarrays. Significant Activities that Support Special Target Populations The objective of this agreement is to establish the proof-of-concept that floating glycoprotein film-coated micelles can be used to capture pathogens from aqueous matrices and stabilize them during transport and storage. Virtually all bacteria possess one or more fimbriae or pili, hair-like structures extending from the cell surface. These fimbriae/pili allow for attachment to various surfaces, including a variety of inert surfaces. Fimbriae/pili associated with pathogens possess adhesions that are responsible for specific attachment to receptor molecules found on human tissues, typically at the site of infection. For example, uropathogenic E. coli (UPEC) possess Type I fimbriae that mediate attachment to mannose-containing glycoproteins coating the urinary tract. The role of Mitre is to provide glycoprotein- coated micelles for experimentation and to provide expertise on adhesion- receptor binding. Meetings are conducted monthly.
Impacts
(N/A)
Publications
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