Community-Based Approaches to Reducing Health Inequities and Fostering Environmental Justice through Global Youth-Engaged Citizen Science

Growing socioeconomic and structural disparities within and between nations have created unprecedented health inequities that have been felt most keenly among the world’s youth. While policy approaches can help to mitigate such inequities, they are often challenging to enact in under-resourced and marginalized communities. Community-engaged participatory action research provides an alternative or complementary means for addressing the physical and social environmental contexts that can impact health inequities. The purpose of this article is to describe the application of a particular form of technology-enabled participatory action research, called the Our Voice citizen science research model, with youth. An overview of 20 Our Voice studies occurring across five continents indicates that youth and young adults from varied backgrounds and with interests in diverse issues affecting their communities can participate successfully in multiple contributory research processes, including those representing the full scientific endeavor. These activities can, in turn, lead to changes in physical and social environments of relevance to health, wellbeing, and, at times, climate stabilization. The article ends with future directions for the advancement of this type of community-engaged citizen science among young people across the socioeconomic spectrum

Socializing One Health: an innovative strategy to investigate social and behavioral risks of emerging viral threats

In an effort to strengthen global capacity to prevent, detect, and control infectious diseases in animals and people, the United States Agency for International Development’s (USAID) Emerging Pandemic Threats (EPT) PREDICT project funded development of regional, national, and local One Health capacities for early disease detection, rapid response, disease control, and risk reduction. From the outset, the EPT approach was inclusive of social science research methods designed to understand the contexts and behaviors of communities living and working at human-animal-environment interfaces considered high-risk for virus emergence. Using qualitative and quantitative approaches, PREDICT behavioral research aimed to identify and assess a range of socio-cultural behaviors that could be influential in zoonotic disease emergence, amplification, and transmission. This broad approach to behavioral risk characterization enabled us to identify and characterize human activities that could be linked to the transmission dynamics of new and emerging viruses. This paper provides a discussion of implementation of a social science approach within a zoonotic surveillance framework. We conducted in-depth ethnographic interviews and focus groups to better understand the individual- and community-level knowledge, attitudes, and practices that potentially put participants at risk for zoonotic disease transmission from the animals they live and work with, across 6 interface domains. When we asked highly-exposed individuals (ie. bushmeat hunters, wildlife or guano farmers) about the risk they perceived in their occupational activities, most did not perceive it to be risky, whether because it was normalized by years (or generations) of doing such an activity, or due to lack of information about potential risks. Integrating the social sciences allows investigations of the specific human activities that are hypothesized to drive disease emergence, amplification, and transmission, in order to better substantiate behavioral disease drivers, along with the social dimensions of infection and transmission dynamics. Understanding these dynamics is critical to achieving health security--the protection from threats to health-- which requires investments in both collective and individual health security. Involving behavioral sciences into zoonotic disease surveillance allowed us to push toward fuller community integration and engagement and toward dialogue and implementation of recommendations for disease prevention and improved health security

Mechanistic Investigations of Cyclase Enzymes Involved in Lantibiotic Biosynthesis

136 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2005.Lantibiotics are peptide antibiotics that contain thioether bridges, termed lanthionines, putatively generated by Michael addition of cysteines to dehydrated serine and threonine residues in the pre-lantibiotic peptide. In class AI lantibiotics, the post-translational modifications are proposed to involve a multi-enzyme complex including the proteins LanB and LanC that are putatively attributed the functions of dehydration and cyclization, respectively, while class AII use a single enzyme, LanM, to carry out both transformations. Previous work in our lab has shown that the LanC enzymes involved in both subtilin and nisin biosynthesis, SpaC and NisC, respectively, contain stoichiometric amounts of zinc, implying a possible catalytic role. EXAFS analysis of SpaC suggests that two cysteines coordinate to the metal and possibly two histidines. The metal may function to activate the cysteine thiol of the substrate toward intramolecular Michael addition to the dehydroalanine and dehydrobutyrine residues in the peptide. Sequence alignments of both the LanC and LanM families of enzymes reveal four conserved residues, two cysteines and two histidines. Mutants of SpaC replacing the conserved cysteines with alanines were generated and characterization by both EXAFS and ICP analysis has shown that these cysteines are involved in zinc binding. Similar studies have been performed using LctM, the LanM protein involved in lacticin 481 biosynthesis. Cysteine and histidine mutants have been generated and analyzed to determine if eliminating the protein's ability to bind zinc leads to a loss in cyclization activity. Results from these experiments implicate Cys836 and His725 in the cyclization mechanism, and yielded mutant enzymes only capable of dehydrating the LctA substrate. In an attempt to understand the structural requirements for synthetase activity, chimeras consisting of the LctA leader sequence fused to structural regions of various class AI and AII lantibiotics have also been generated and treated with LctM to test for activity.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

Mechanistic Investigations of Cyclase Enzymes Involved in Lantibiotic Biosynthesis

136 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2005.Lantibiotics are peptide antibiotics that contain thioether bridges, termed lanthionines, putatively generated by Michael addition of cysteines to dehydrated serine and threonine residues in the pre-lantibiotic peptide. In class AI lantibiotics, the post-translational modifications are proposed to involve a multi-enzyme complex including the proteins LanB and LanC that are putatively attributed the functions of dehydration and cyclization, respectively, while class AII use a single enzyme, LanM, to carry out both transformations. Previous work in our lab has shown that the LanC enzymes involved in both subtilin and nisin biosynthesis, SpaC and NisC, respectively, contain stoichiometric amounts of zinc, implying a possible catalytic role. EXAFS analysis of SpaC suggests that two cysteines coordinate to the metal and possibly two histidines. The metal may function to activate the cysteine thiol of the substrate toward intramolecular Michael addition to the dehydroalanine and dehydrobutyrine residues in the peptide. Sequence alignments of both the LanC and LanM families of enzymes reveal four conserved residues, two cysteines and two histidines. Mutants of SpaC replacing the conserved cysteines with alanines were generated and characterization by both EXAFS and ICP analysis has shown that these cysteines are involved in zinc binding. Similar studies have been performed using LctM, the LanM protein involved in lacticin 481 biosynthesis. Cysteine and histidine mutants have been generated and analyzed to determine if eliminating the protein's ability to bind zinc leads to a loss in cyclization activity. Results from these experiments implicate Cys836 and His725 in the cyclization mechanism, and yielded mutant enzymes only capable of dehydrating the LctA substrate. In an attempt to understand the structural requirements for synthetase activity, chimeras consisting of the LctA leader sequence fused to structural regions of various class AI and AII lantibiotics have also been generated and treated with LctM to test for activity.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

Direct, quantitative detection of Listeria monocytogenes in fresh raw whole milk by qPCR

A method previously developed for direct (non-enrichment) detection of Escherichia coli O157:H7 was adapted for Listeria monocytogenes. The sample treatment protocol was successful in concentrating bacteria from 10 mL raw milk samples and reducing PCR inhibition, but qPCR detection sensitivity and reproducibility was poor. Two DNA extraction reagents and multiple extraction conditions were tested to identify an efficient and reproducible DNA extraction method. Two primer/probe sets were evaluated at two concentrations and three annealing temperatures to minimize false-positive results and optimize sensitivity and reproducibility of qPCR detection. Under the selected conditions, DNA was extracted efficiently from the entire milk sample in a volume of 10 mL, and subsequently quantitated by a 50 nuclease qPCR assay lasting 50 min. The method provided detection of 1 cfu mL1 L. monocytogenes in 10 mL raw milk and quantitation from 10 to 1000 cfu mL1 with a total time to result of <3 h

Two Methods for Increased Specificity and Sensitivity in Loop-Mediated Isothermal Amplification

The technique of loop-mediated isothermal amplification (LAMP) utilizes four (or six) primers targeting six (or eight) regions within a fairly small segment of a genome for amplification, with concentration higher than that used in traditional PCR methods. The high concentrations of primers used leads to an increased likelihood of non-specific amplification induced by primer dimers. In this study, a set of LAMP primers were designed targeting the prfA gene sequence of Listeria monocytogenes, and dimethyl sulfoxide (DMSO) as well as Touchdown LAMP were employed to increase the sensitivity and specificity of the LAMP reactions. The results indicate that the detection limit of this novel LAMP assay with the newly designed primers and additives was 10 fg per reaction, which is ten-fold more sensitive than a commercial Isothermal Amplification Kit and hundred-fold more sensitive than previously reported LAMP assays. This highly sensitive LAMP assay has been shown to detect 11 strains of Listeria monocytogenes, and does not detect other Listeria species (including Listeria innocua and Listeria invanovii), providing some advantages in specificity over commercial Isothermal Amplification Kits and previously reported LAMP assay