The Commit to Be Fit framework: a community case study of a multi-level, holistic school-based wellness initiative in rural Virginia.

Background: Public health interventions that target children's physical, mental, and emotional health will enhance their ability to learn and grow. Although more complex, school initiatives that address multiple ecological levels and take a holistic view may be more effective and likely to lead to lasting change. Aims: This article presents the framework of Commit to Be Fit (C2BF) as an example of how schools can integrate multi-level and holistic approaches for health. This innovative school-based intervention includes activities addressing individual, home, school, and community to create a culture of wellness. We describe the implementation of C2BF and its basis in ecological models and give examples of activities across three components: cafeteria, classroom, and community. We discuss challenges and note that leadership engagement and alignment were critical elements for C2BF's success thus far. Discussion: C2BF uses a school-based multi-level approach to creating a culture of wellness and holistic health for students, teachers, and community members. C2BF is unique compared to other school-based programming and includes activities that address all eight domains posited for program sustainability within public health. Built to be flexible and adaptive, C2BF was able to successfully pivot during the COVID pandemic and also follow new science. Conclusion: C2BF and other multi-level holistic approaches are more likely to achieve long-term change by utilizing strategies across the multiple levels of the ecological model to improve health and wellbeing.

The Small Metal-Binding Protein SmbP Simplifies the Recombinant Expression and Purification of the Antimicrobial Peptide LL-37.

(1) Background: The cathelicidin peptide LL-37 is a prominent molecule with many biological activities, including antimicrobial. Due to its importance, here, we describe the production of LL-37 tagged with SmbP, a relatively new carrier protein that improves the production of recombinant proteins and peptides in Escherichia coli. We present an alternative method for the rapid expression, purification, and antimicrobial evaluation of LL-37, that involves only one purification step. (2) Methods: A DNA construct of SmbP_LL-37 was transformed into E. coli BL21(DE3); after overnight expression, the protein was purified directly from the cell lysate using immobilized metal-affinity chromatography. SmbP_LL-37 was treated with Enterokinase to obtain the free LL-37 peptide. The antimicrobial activity of both SmbP_LL-37 and free LL-37 was determined using the colony forming unit assay method. (3) Results: SmbP_LL-37 was observed in the soluble fraction of the cell lysate; after purification with IMAC, protein gel electrophoresis, and analysis by ImageJ, it showed 90% purity. A total of 3.6 mg of SmbP_LL-37 was produced from one liter of cell culture. SmbP_LL-37 and free LL-37 both showed inhibition activity against Staphylococcus aureus and Escherichia coli. (4) Conclusions: The SmbP fusion protein is a valuable tool for producing biologically-active LL-37 peptide. The production method described here should be of interest for the expression and purification of additional cationic peptides, since it cuts the purification time considerably prior to determination of antimicrobial activity.

Expression and Purification of Recombinant Proteins in Escherichia coli Tagged with the Metal-Binding Proteins SmbP and CusF3H.

The bacterium Escherichia coli is still considered the first option as a microbial cell factory for recombinant protein production, and affinity chromatography is by far the preferred technique for initial purification after protein expression and cell lysis. In this chapter, we describe the methodology to express and purify recombinant proteins in E. coli tagged with the first two metal-binding proteins proposed as fusion partners. They are the small metal-binding protein SmbP and a mutant of the copper resistance protein CusF3H+. There are several advantages of using them as protein tags: they prevent the formation of inclusion bodies by increasing solubility of the target proteins, they enable purification by immobilized metal-affinity chromatography using Ni(II) ions with high purity, and because of their low molecular weights, excellent final yields are obtained for the target proteins after cleavage and removal of the protein tag. Here we also describe the protocol for the production of proteins in the periplasm of E. coli tagged with two SmbP variants that include the PelB or the TorA signal sequences for transport via the Sec or the Tat pathway, respectively. Based on these methods, we consider CusF3H+ and SmbP excellent alternatives as fusion proteins for the production of recombinant proteins in E. coli.

Engineered small metal-binding protein tag improves the production of recombinant human growth hormone in the periplasm of Escherichia coli.

Fusion proteins play an important role in the production of recombinant proteins in Escherichia coli. They are mostly used for cytoplasmic expression since they can be designed to increase the solubility of the target protein, which then can be easily purified via affinity chromatography. In contrast, fusion proteins are not usually included in construct designs for periplasmic production. Instead, a signal sequence is inserted for protein transport into the periplasm and a C-terminal his-tag added for subsequent purification. Our research group has proposed the small metal-binding protein (SmbP) isolated from the periplasm of Nitrosomonas europaea as a new fusion protein to express recombinant proteins in the cytoplasm or periplasm of E. coli. SmbP also allows purification via immobilized metal affinity chromatography using Ni(II) ions. Recently, we have optimized the periplasmic production of proteins tagged with SmbP by exchanging its native signal peptide with one taken from pectate lyase B (PelB), substantially increasing the amount of protein produced. In this work, we have expressed and purified soluble bioactive human growth hormone (hGH) tagged with PelB-SmbP and obtained the highest periplasmic production reported for this protein so far. Its activity, tested on Nb2-11 cells, was equivalent to commercial growth hormone at 50 ng·mL-1 . Therefore, we strongly recommend the use of PelB-SmbP as a protein tag for the expression and purification of hGH or other possible target proteins in the periplasm of E. coli.

Modos de vida y estado de salud de migrantes en un asentamiento de Barranquilla, 2018 / Lifestyles and health status of migrants in a settlement of Barranquilla, Colombia, 2018

RESUMEN Objetivo Describir los modos de vida y el estado de salud de salud de migrantes venezolanos y colombianos de retorno asentados en Villa Caracas, Barranquilla, en el año 2018. Método Estudio descriptivo de corte transversal con muestreo sistemático de viviendas. Fueron incluidas 229 personas mayores de 15 años procedentes de 90 viviendas. Resultados Se encontraron diferencias en las rutas para llegar, tiempos de traslado y estancia en el asentamiento entre migrantes venezolanos y colombianos en retorno. Las condiciones de la vivienda y el acceso a los servicios públicos son limitadas, menos de la mitad de las viviendas tienen acceso a acueducto, alcantarillado y baño. En general el estado de salud auto-reportado por los migrantes es muy bueno o bueno, las prevalencias de enfermedades crónicas fueron relativamente bajas, con excepción de hipertensión arterial. De los que consultaron al servicio de urgencias, la mayoría reportó acceso efectivo. Se encontraron síntomas depresivos clínicamente significativos para el 20% de la población encuestada. Conclusiones Los migrantes de Villa Caracas se encuentran en condiciones de alta vulnerabilidad social dadas sus condiciones económicas y ambientales. A pesar de la falta de aseguramiento al sistema de salud colombiano, reportaron acceso a la atención por urgencias.(AU)

ABSTRACT Objective To describe the lifestyles and health status of returning Venezuelan and Colombian migrants in Villa Caracas, Barranquilla, in 2018. Methods Descriptive, cross-sectional study with systematic sampling of dwellings. 229 people over 15 years of age from 90 homes were included. Results Differences were found in the routes to arrive, commuting times and stay in the settlement between Venezuelan and returning Colombian migrants. Housing conditions and access to public services are limited: less than half of the dwellings have access to aqueduct, sewerage and bathrooms. In general, self-reported health status of migrants is very good or good and the prevalence of noncommunicable diseases was relatively low, with the exception of high blood pressure. Most of the people who consulted the emergency department reported effective access. Clinically significant depressive symptoms were found in 20% of the surveyed population. Conclusions The migrants of Villa Caracas are under high social vulnerability conditions given their economic and environmental conditions. Despite their lack of enrollment in the Colombian health system, they reported access to emergency care.(AU)

[Lifestyles and health status of migrants in a settlement of Barranquilla, Colombia, 2018]. / Modos de vida y estado de salud de migrantes en un asentamiento de Barranquilla, 2018.

OBJECTIVE: To describe the lifestyles and health status of returning Venezuelan and Colombian migrants in Villa Caracas, Barranquilla, in 2018. METHODS: Descriptive, cross-sectional study with systematic sampling of dwellings. 229 people over 15 years of age from 90 homes were included. RESULTS: Differences were found in the routes to arrive, commuting times and stay in the settlement between Venezuelan and returning Colombian migrants. Housing conditions and access to public services are limited: less than half of the dwellings have access to aqueduct, sewerage and bathrooms. In general, self-reported health status of migrants is very good or good and the prevalence of noncommunicable diseases was relatively low, with the exception of high blood pressure. Most of the people who consulted the emergency department reported effective access. Clinically significant depressive symptoms were found in 20% of the surveyed population. CONCLUSIONS: The migrants of Villa Caracas are under high social vulnerability conditions given their economic and environmental conditions. Despite their lack of enrollment in the Colombian health system, they reported access to emergency care.

OBJETIVO: Describir los modos de vida y el estado de salud de salud de migrantes venezolanos y colombianos de retorno asentados en Villa Caracas, Barranquilla, en el año 2018. MÉTODO: Estudio descriptivo de corte transversal con muestreo sistemático de viviendas. Fueron incluidas 229 personas mayores de 15 años procedentes de 90 viviendas. RESULTADOS: Se encontraron diferencias en las rutas para llegar, tiempos de traslado y estancia en el asentamiento entre migrantes venezolanos y colombianos en retorno. Las condiciones de la vivienda y el acceso a los servicios públicos son limitadas, menos de la mitad de las viviendas tienen acceso a acueducto, alcantarillado y baño. En general el estado de salud auto-reportado por los migrantes es muy bueno o bueno, las prevalencias de enfermedades crónicas fueron relativamente bajas, con excepción de hipertensión arterial. De los que consultaron al servicio de urgencias, la mayoría reportó acceso efectivo. Se encontraron síntomas depresivos clínicamente significativos para el 20% de la población encuestada. CONCLUSIONES: Los migrantes de Villa Caracas se encuentran en condiciones de alta vulnerabilidad social dadas sus condiciones económicas y ambientales. A pesar de la falta de aseguramiento al sistema de salud colombiano, reportaron acceso a la atención por urgencias.

Correlation between diet and gut bacteria in a population of young adults.

Dietary habits strongly influence gut microbiota. The aim of this study was to compare and correlated the abundance of Firmicutes and Bacteroidetes phyla, some representative bacteria of these phyla such as Bacteroides thetaiotaomicron, Prevotella, Faecalibacterium prausnitzii, Clostridium leptum and Bifidobacterium longum as a member of Actinobacteria phylum in young adults with their food intake. Faecal samples used came from lean subjects (BMI = 19.83 ± 0.94 kg/m(2)), overweight (BMI = 27.17 ± 0.51 kg/m(2)) and obese (BMI = 41.33 ± 5.25 kg/m(2)). There were significant differences in total studied gut microbiota between the overweight and lean groups. Members of the Firmicutes phylum, and Bifidobacterium longum, were more abundant in the lean group. The results suggest that diet rich in unsaturated fatty acids and fibre promote an abundant population of beneficial bacteria such as B. longum and Bacteroidetes. However, it has been considered that the results may be biased due to the size of the individuals studied; therefore the results could be only valid for the studied population.

A Simple Mathematical Model for Wound Closure Evaluation.

The incidence of ulcers associated to type 2 diabetes mellitus (T2DM) increases every year. We introduce and explore a new mathematical algorithm to evaluate wound-healing in foot ulcers associated to T2DM. Fifteen patients (nine women and six men), mean age of 70 ± 16 years were included. The evolution of their wounds followed-up for a period of 18-45 days. According to the Wagner grading system the ulcers were grade I (5 patients), grade II (9 patients), and grade III (1 patient). Clinically, the type of the ulcers was neuroischemic (12 patients) and neuropathic (3 patients). A new parameter is introduced, the "continuous linear healing rate" Dc that was more accurate with higher values and requires less quantifications than usual formulas to make a wound-healing projection.

Protein clustering and RNA phylogenetic reconstruction of the influenza A [corrected] virus NS1 protein allow an update in classification and identification of motif conservation.

The non-structural protein 1 (NS1) of influenza A virus (IAV), coded by its third most diverse gene, interacts with multiple molecules within infected cells. NS1 is involved in host immune response regulation and is a potential contributor to the virus host range. Early phylogenetic analyses using 50 sequences led to the classification of NS1 gene variants into groups (alleles) A and B. We reanalyzed NS1 diversity using 14,716 complete NS IAV sequences, downloaded from public databases, without host bias. Removal of sequence redundancy and further structured clustering at 96.8% amino acid similarity produced 415 clusters that enhanced our capability to detect distinct subgroups and lineages, which were assigned a numerical nomenclature. Maximum likelihood phylogenetic reconstruction using RNA sequences indicated the previously identified deep branching separating group A from group B, with five distinct subgroups within A as well as two and five lineages within the A4 and A5 subgroups, respectively. Our classification model proposes that sequence patterns in thirteen amino acid positions are sufficient to fit >99.9% of all currently available NS1 sequences into the A subgroups/lineages or the B group. This classification reduces host and virus bias through the prioritization of NS1 RNA phylogenetics over host or virus phenetics. We found significant sequence conservation within the subgroups and lineages with characteristic patterns of functional motifs, such as the differential binding of CPSF30 and crk/crkL or the availability of a C-terminal PDZ-binding motif. To understand selection pressures and evolution acting on NS1, it is necessary to organize the available data. This updated classification may help to clarify and organize the study of NS1 interactions and pathogenic differences and allow the drawing of further functional inferences on sequences in each group, subgroup and lineage rather than on a strain-by-strain basis.