Bioinformatics-guided discovery of biaryl-linked lasso peptides.

Lasso peptides are a class of ribosomally synthesized and post-translationally modified peptides (RiPPs) that feature an isopeptide bond and a distinct lariat fold. A growing number of secondary modifications have been described that further decorate lasso peptide scaffolds. Using genome mining, we have discovered a pair of lasso peptide biosynthetic gene clusters (BGCs) that include cytochrome P450 genes. Using mass spectrometry, stable isotope incorporation, and extensive 2D-NMR spectrometry, we report the structural characterization of two unique examples of (C-N) biaryl-linked lasso peptides. Nocapeptin A, from Nocardia terpenica, is tailored with a Trp-Tyr crosslink, while longipepetin A, from Longimycelium tulufanense, features a Trp-Trp linkage. Besides the unusual bicyclic frame, a Met of longipepetin A undergoes S-methylation to yield a trivalent sulfonium, a heretofore unprecedented RiPP modification. A bioinformatic survey revealed additional lasso peptide BGCs containing P450 enzymes which await future characterization. Lastly, nocapeptin A bioactivity was assessed against a panel of human and bacterial cell lines with modest growth-suppression activity detected towards Micrococcus luteus.

A Unique Model and Workforce to Address Health-Related Social Needs and Health Equity: Regional Health Connectors in Colorado.

Unmet health-related social needs contribute to high morbidity and poor population health. Improving social conditions are likely to reduce health disparities and improve the health of the overall U.S. population. The primary objective of this article is to describe an innovative workforce model, called Regional Health Connectors (RHCs), and how they address health-related social needs in Colorado. This is a program evaluation that analyzed field notes and interview data from 2021-2022. We applied our findings to the framework developed by the National Academies of Sciences, Engineering, and Medicine's (NASEM's) report on strengthening social care integration into health care (2019). We found that RHCs address the following health-related social needs most commonly: food insecurity (n = 18 of 21 regions or 85% of all regions), housing (n = 17 or 81% of all regions), transportation (n = 11 or 52% of all regions), employment opportunities (n = 10 or 48% of all regions), and income/financial assistance (n = 11 or 52% of all regions). RHCs interacted across many sectors to address health-related social needs and provided multiple types of support to primary care practices at the organizational level. Examples of emerging impact of RHCs are described and mapped onto the NASEM framework. Findings from this program evaluation add to the growing landscape of knowledge and importance of detecting and addressing health-related social needs. We conclude that RHCs are a unique and emerging workforce that addresses multiple domains needed to integrate social care into health care.

Bioinformatics-Guided Discovery of Biaryl-Tailored Lasso Peptides.

Lasso peptides are a class of ribosomally synthesized and post-translationally modified peptides (RiPPs) that feature an isopeptide bond and a distinct lariat fold. A growing number of secondary modifications have been described that further decorate lasso peptide scaffolds. Using genome mining, we have discovered a pair of lasso peptide biosynthetic gene clusters (BGCs) that include cytochrome P450 genes. Here, we report the structural characterization of two unique examples of (C-N) biaryl-containing lasso peptides. Nocapeptin A, from Nocardia terpenica, is tailored with Trp-Tyr crosslink while longipepetin A, from Longimycelium tulufanense, features Trp-Trp linkage. Besides the unusual bicyclic frame, longipepetin A receives an S-methylation by a new Met methyltransferase resulting in unprecedented sulfonium-bearing RiPP. Our bioinformatic survey revealed P450(s) and further maturating enzyme(s)-containing lasso BGCs awaiting future characterization.

Evolution of Artificial Arginine Analogues-Fluorescent Guanidiniocarbonyl-Indoles as Efficient Oxo-Anion Binders.

In this article, we present fluorescent guanidiniocarbonyl-indoles as versatile oxo-anion binders. Herein, the guanidiniocarbonyl-indole (GCI) and methoxy-guanidiniocarbonyl-indole (MGCI) were investigated as ethylamides and compared with the well-known guanidiniocarbonyl-pyrrole (GCP) concerning their photophysical properties as well as their binding behavior towards oxo-anions. Hence, a variety of anionic species, such as carboxylates, phosphonates and sulfonates, have been studied regarding their binding properties with GCP, GCI and MGCI using UV-Vis titrations, in combination with the determination of the complex stoichiometry using the Job method. The emission properties were studied in relation to the pH value using fluorescence spectroscopy as well as the determination of the photoluminescence quantum yields (PLQY). Density functional theory (DFT) calculations were undertaken to obtain a better understanding of the ground-lying electronic properties of the investigated oxo-anion binders. Additionally, X-ray diffraction of GCP and GCI was conducted. We found that GCI and MGCI efficiently bind carboxylates, phosphonates and sulfonates in buffered aqueous solution and in a similar range as GCP (Kass ≈ 1000-18,000 M-1, in bis-tris buffer, pH = 6); thus, they could be regarded as promising emissive oxo-anion binders. They also exhibit a visible fluorescence with a sufficient PLQY. Additionally, the excitation and emission wavelength of MGCI was successfully shifted closer to the visible region of the electromagnetic spectrum by introducing a methoxy-group into the core structure, which makes them interesting for biological applications.

Development and Scoring of a Survey on Public Health Accreditation Capacity.

OBJECTIVES: Public health accreditation is a 7-step process that starts with a period of preapplication during which a health department assesses its readiness for accreditation. However, no tools with established reliability and validity that quantitatively measure a local health department's (LHD's) capacity for accreditation are available to complete this initial step. We developed and validated a survey to measure accreditation capacity for LHDs. METHODS: From January through April 2015, we administered a cross-sectional electronic survey instrument with 15 questions that tapped into domains of capacity for public health accreditation. We analyzed and grouped responses by using a confirmatory maximum likelihood factor analysis with oblique rotations. We assessed reliability by using Cronbach α, and we assessed validity by comparing responses with previously established instruments. We administered the survey to 174 LHD directors in Colorado, Kansas, and Nebraska, 153 (88%) of whom responded. RESULTS: The factor analysis produced a 3-factor model of accreditation capacity, suggesting that accreditation capacity depends on 3 distinct latent constructs: support for accreditation, preparation, and planning and approach. The model had good scale reliability (average Cronbach α = 0.7) and validity (average factor correlation = 0.43). CONCLUSIONS: The survey developed and scored in this analysis can be used by LHDs to inform the feasibility of initiating the time-intensive and costly process of accreditation.

The Economic Cost of Communicable Disease Surveillance in Local Public Health Agencies.

OBJECTIVE: We identify economic costs associated with communicable disease (CD) monitoring/surveillance in Colorado local public health agencies and identify possible economies of scale. DATA SOURCES/STUDY SETTING: Data were collected via a survey of local public health employees engaged in CD work. Survey respondents logged time spent on CD surveillance for 2-week periods in the spring of 2014 and fall of 2014. Forty-three of the 54 local public health agencies in Colorado participated. STUDY DESIGN: We used a microcosting approach. We estimated a statistical cost function using cost as a function of the number of reported investigable diseases during the matched 2-week period. We also controlled for other independent variables, including case mix, characteristics of the agency, the community, and services provided. DATA COLLECTION/EXTRACTION METHODS: Data were collected from a microcosting survey using time logs. PRINCIPAL FINDINGS: Costs increased at a decreasing rate as cases increased, with both cases (ß = 431.5, p < .001) and cases squared (ß = -3.62, p = .05) statistically significant. CONCLUSIONS AND IMPLICATIONS: The results of the model suggest economies of scale. Cost per unit is estimated to be one-third lower for high-volume agencies as compared to low-volume agencies. Cost savings could potentially be achieved if smaller agencies shared services.

Strategic Methodologies in Public Health Cost Analyses.

CONTEXT: The National Research Agenda for Public Health Services and Systems Research states the need for research to determine the cost of delivering public health services in order to assist the public health system in communicating financial needs to decision makers, partners, and health reform leaders. OBJECTIVE: The objective of this analysis is to compare 2 cost estimation methodologies, public health manager estimates of employee time spent and activity logs completed by public health workers, to understand to what degree manager surveys could be used in lieu of more time-consuming and burdensome activity logs. DESIGN: Employees recorded their time spent on communicable disease surveillance for a 2-week period using an activity log. Managers then estimated time spent by each employee on a manager survey. Robust and ordinary least squares regression was used to measure the agreement between the time estimated by the manager and the time recorded by the employee. MAIN OUTCOME MEASURES: The 2 outcomes for this study included time recorded by the employee on the activity log and time estimated by the manager on the manager survey. SETTING: This study was conducted in local health departments in Colorado. PARTICIPANTS: Forty-one Colorado local health departments (82%) agreed to participate. RESULTS: Seven of the 8 models showed that managers underestimate their employees' time, especially for activities on which an employee spent little time. Manager surveys can best estimate time for time-intensive activities, such as total time spent on a core service or broad public health activity, and yet are less precise when estimating discrete activities. CONCLUSIONS: When Public Health Services and Systems Research researchers and health departments are conducting studies to determine the cost of public health services, there are many situations in which managers can closely approximate the time required and produce a relatively precise approximation of cost without as much time investment by practitioners.

Minimum package of public health services: the adoption of core services in local public health agencies in Colorado.

OBJECTIVES: We examined the effect of a state law in Colorado that required local public health agencies to deliver a minimum package of public health services. METHODS: We used a longitudinal, pre-post study design, with baseline data collected in 2011 and follow-up data collected in 2013. We conducted means testing to analyze the change in service delivery and activities. We conducted linear regression to test for system structure effects on the implementation of core services. RESULTS: We observed statistically significant increases in several service areas within communicable disease, prevention and population health promotion, and environmental health. In addition to service and program areas, specific activities had significant increases. The significant activity increases were all in population- and systems-based services. CONCLUSIONS: This project provided insight into the likely effect of national adoption of a minimum package as recommended by the Institute of Medicine. The implementation of a minimum package showed significant changes in service delivery, with specific service delivery measurement over a short period of time. Our research sets up a research framework to further explore core service delivery measure development.

An examination of state laws and policies regarding public health agency accreditation prerequisites.

CONTEXT: The Public Health Accreditation Board (PHAB) began accepting applications for national voluntary accreditation in the fall of 2011. There are 3 prerequisites for accreditation: health assessments; health improvement plans; and strategic plans. These prerequisites must be in place for a state or local public health agency to apply for PHAB accreditation. Completion of the prerequisites can become a barrier for agencies that are considering applying for accreditation. OBJECTIVE: This project looked at all 50 states through a legal lens to identify legal tools in the form of laws, rules, executive orders, contracts, legislative resolutions, and other tools that are used to authorize or require that a state or local health agency complete 1 or more of the prerequisites. DESIGN: Using legal research platforms and a Boolean search stream, an inquiry of legal tools in all 50 states was completed to determine where legal tools are used to authorize or require the 3 PHAB prerequisites. Once legal tools were found, interviews were conducted with key informants from each state to understand the implementation and use of the legal tools found and collect any additional legal tools that were not found from the legal search. RESULTS: The results include key findings and, importantly, an inventory of laws and legal tools that are being used around the country to require and support completion of the prerequisites. Within all 50 states, 26 states have some type of a mandate regarding 1 or more of the PHAB prerequisites. This includes 1 state that includes a mandate for PHAB accreditation. CONCLUSIONS: States use a wide variety of legal tools to implement the prerequisites for voluntary accreditation. It is important to understand the interpretation, enforcement, and support of the laws and legal tools to determine whether the tools have impact in individual states.

An assessment of schoolyard features and behavior patterns in children's utilization and physical activity.

BACKGROUND: Careful research that elucidates how behavior relates to design in the context of elementary school grounds can serve to guide cost-efficient design with the goal of encouraging physical activity (PA). This work explores patterns in children's PA behavior within playground spaces with the specific goal of guiding healthy playground design. METHODS: Data on children's utilization and PA behavior in 6 playgrounds divided into 106 observation zones were collected in 2005 and 2006 at Denver elementary school playgrounds using the System for Observing Play and Leisure Activity in Youth. Analyses of variance and t tests determined whether there were differences in utilization and behavior patterns across observations zones and between genders. RESULTS: This study provides evidence that children prefer to use certain types of playground zones and that they are more likely to practice moderate-to-vigorous physical activity (MVPA) in some zones. The authors observed statistically significant differences between genders. Boys were more likely to engage in MVPA in zones without equipment, girls were more likely to use zones with equipment. CONCLUSIONS: This work suggests that the inclusion or omission of specific playground features may have an impact on the way that children use the spaces.

An assessment of schoolyard renovation strategies to encourage children's physical activity.

BACKGROUND: Children in poor and minority neighborhoods often lack adequate environmental support for healthy physical development and community interventions designed to improve physical activity resources serve as an important approach to addressing obesity. In Denver, the Learning Landscapes (LL) program has constructed over 98 culturally-tailored schoolyard play spaces at elementary schools with the goal to encourage utilization of play spaces and physical activity. In spite of enthusiasm about such projects to improve urban environments, little work has evaluated their impact or success in achieving their stated objectives. This study evaluates the impacts of LL construction and recency of renovation on schoolyard utilization and the physical activity rates of children, both during and outside of school, using an observational study design. METHODS: This study employs a quantitative method for evaluating levels of physical activity of individuals and associated environmental characteristics in play and leisure environments. Schools were selected on the basis of their participation in the LL program, the recency of schoolyard renovation, the size of the school, and the social and demographic characteristics of the school population. Activity in the schoolyards was measured using the System for Observing Play and Leisure Activity (SOPLAY), a validated quantitative method for evaluating levels of physical activity of individuals in play and leisure environments. Trained observers collected measurements before school, during school recess, after school, and on weekends. Overall utilization (the total number of children observed on the grounds) and the rate of activity (the percentage of children observed who were physically active) were analyzed. Observations were compared using t-tests and the data were stratified by gender for further analysis. In order to assess the impacts of LL renovation, recently-constructed LL schoolyards were compared to LL schoolyards with older construction, as well as un-renovated schoolyards. RESULTS: Overall utilization was significantly higher at LL schools than at un-renovated schools for most observation periods. Notably, LL renovation had no impact on girl's utilization on the weekends, although differences were observed for all other periods. There were no differences in rates of activity for any comparison. With the exception of the number of boys observed, there was no statistically significant difference in activity when recently-constructed LL schools are compared to LL schools with older construction dates and there was no difference observed in comparisons of older LL with unrenovated sites. CONCLUSIONS: While we observed greater utilization and physical activity in schools with LL, the impact of specific features of LL renovation is not clear. However, schoolyard renovation and programs to encourage schoolyard use before and after school may offer a means to encourage greater physical activity among children, and girls in particular. Additional study of schoolyard renovation may shed light on the specific reasons for these findings or suggest effective policies to improve the physical activity resources of poor and minority neighborhoods.

Influence of schoolyard renovations on children's physical activity: the Learning Landscapes Program.

OBJECTIVES: We examined whether schoolyard improvements led to increased physical activity levels among both boys and girls and assessed the aspects of schoolyard design that have an impact on physical activity. METHODS: In a quasi-experimental research design, 6 schools with renovated schoolyards and 3 control schools were divided into activity areas. We calculated measures of children's physical activity by area during school hours as well as after-school hours. RESULTS: The volume of schoolyard use was significantly higher at schools with renovated schoolyards than at control schools, and students were significantly more active at these schools. Also, activity levels were significantly higher among both boys and girls in certain schoolyard areas, such as those with soft surfaces. CONCLUSIONS: Because few public elementary schools in the United States provide daily physical education or its equivalent for all students throughout the school year, noncurriculum approaches to increasing children's physical activity are important. Renovated schoolyards increase the number of children who are physically active, as well as their overall activity levels, and reduce sedentary behaviors.