STRUCTURAL AND FUNCTIONAL STUDIES OF TAILSPIKE PROTEINS FROM ESCHERICHIA COLI O157:H7 PHAGE CBA120

Bacteriophage CBA120, a member of the Ackermannviridae family, was isolated, sequenced, shown to selectively infect the food pathogen E. coli O157:H7. The CBA120 genome encodes four separate tailspike proteins (TSPs), TSP1-4 corresponding to ORFs 210 through 213. TSPs bind and degrade or modify the lipopolysaccharide (LPS) on the bacterial cell surface as part of the adsorption apparatus of tailed bacteriophages. Electron microscopy revealed that phage CBA120 possesses a long contractible tail with distinct star-like structures attached to the baseplate of the tail. Sequence analysis of the four CBA120 TSPs suggests two distinct classes; one class comprising TSP1 and TSP3 corresponds to two-domain structures. The second class comprising TSP2 and TSP4 have longer amino acid sequences and contain additional N-terminal regions responsible for the assembly of the TSP star-like complex. Crystal structure of TSP1 at 1.8 Å resolution and TSP3 at 1.85 Å resolution revealed two-domain homotrimers displaying conserved N-terminal head domain structures and C-terminal receptor binding domains with an overall β-helical fold. Sequence analysis of the assembly regions of TSP2 and TSP4 reveal remote homology to gp10 of phage T4, which is involved in the assembly of the phage baseplate. The crystal structure of TSP2 reveals the head domain and receptor binding domain, but the N-terminal assembly region is structurally disordered. In contrast, the assembly region of TSP4 has been purified without the head and receptor domain; pull-down experiments showed that this region binds TSP1. Bacterial halo assays of TSPs 1-3 revealed that all three proteins produced circles of clearing corresponding to polysaccharide depolymerase activity. Site-directed mutagenesis coupled with the halo assay confirmed the identity of TSP3’s catalytic residues as Asp383 and Asp426, which, based on the structure, suggests that the enzyme employs an acid/base mechanism to degrade lipopolysaccharide. The structure-based putative active site of TSP2 suggests that the catalytic machinery comprises Asp506 and Asp571. It was also shown that TSP2, but not TSP1 or TSP3, diminishes CBA120’s ability to infect E. coli O157:H7. Thus, TSP2 is the phage specificity determinant that binds to the lipopolysaccharide of E. coli O157:H7, whereas the bacterial targets of TSP1 and TSP3 remain unknown

Predictors of drinking and functional outcomes for men and women following inpatient alcohol treatment

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106952/1/ajad12098.pd

Food composition tables in resource-poor settings: Exploring current limitations and opportunities, with a focus on animal-source foods in sub-Saharan Africa

Animal-source foods (ASF) have the potential to enhance the nutritional adequacy of cereal-based diets in low- and middle-income countries, through the provision of high-quality protein and bioavailable micronutrients. The development of guidelines for including ASF in local diets requires an understanding of the nutrient content of available resources. This article reviews food composition tables (FCT) used in sub-Saharan Africa, examining the spectrum of ASF reported and exploring data sources for each reference. Compositional data are shown to be derived from a small number of existing data sets from analyses conducted largely in high-income nations, often many decades previously. There are limitations in using such values, which represent the products of intensively raised animals of commercial breeds, as a reference in resource-poor settings where indigenous breed livestock are commonly reared in low-input production systems, on mineral-deficient soils and not receiving nutritionally balanced feed. The FCT examined also revealed a lack of data on the full spectrum of ASF, including offal and wild foods, which correspond to local food preferences and represent valuable dietary resources in food-deficient settings. Using poultry products as an example, comparisons are made between compositional data from three high-income nations, and potential implications of differences in the published values for micronutrients of public health significance, including Fe, folate and vitamin A, are discussed. It is important that those working on nutritional interventions and on developing dietary recommendations for resource-poor settings understand the limitations of current food composition data and that opportunities to improve existing resources are more actively explored and supported

General population screening for type 1 diabetes using islet autoantibodies at the preschool vaccination visit:a proof-of-concept study (the T1Early study)

Objective: Type 1 diabetes (T1D) screening programmes testing islet autoantibodies (IAbs) in childhood can reduce life-threatening diabetic ketoacidosis. General population screening is required to detect the majority of children with T1D, since in >85% there is no family history. Age 3–5 years has been proposed as an optimal age for a single screen approach. Design: Capillary samples were collected from children attending their preschool vaccination and analysed for IAbs to insulin, glutamic acid decarboxylase, islet antigen-2 and zinc transporter 8 using radiobinding/luciferase immunoprecipitation system assays. Acceptability was assessed using semistructured interviews and open-ended postcard questionnaires with parents. Setting: Two primary care practices in Oxfordshire, UK. Main outcome measures: The ability to collect capillary blood to test IAbs in children at the routine preschool vaccination (3.5–4 years). Results: Of 134 parents invited, 66 (49%) were recruited (median age 3.5 years (IQR 3.4–3.6), 26 (39.4%) male); 63 provided a sample (97% successfully), and one participant was identified with a single positive IAb. Parents (n=15 interviews, n=29 postcards) were uniformly positive about screening aligned to vaccination and stated they would have been less likely to take part had screening been a separate visit. Themes identified included preparedness for T1D and the long-term benefit outweighing short-term upset. The perceived volume of the capillary sample was a potential concern and needs optimising. Conclusions: Capillary IAb testing is a possible method to screen children for T1D. Aligning collection to the preschool vaccination visit can be convenient for families without the need for an additional visit

General population screening for type 1 diabetes using islet autoantibodies at the preschool vaccination visit:a proof-of-concept study (the T1Early study)

Objective: Type 1 diabetes (T1D) screening programmes testing islet autoantibodies (IAbs) in childhood can reduce life-threatening diabetic ketoacidosis. General population screening is required to detect the majority of children with T1D, since in >85% there is no family history. Age 3–5 years has been proposed as an optimal age for a single screen approach. Design: Capillary samples were collected from children attending their preschool vaccination and analysed for IAbs to insulin, glutamic acid decarboxylase, islet antigen-2 and zinc transporter 8 using radiobinding/luciferase immunoprecipitation system assays. Acceptability was assessed using semistructured interviews and open-ended postcard questionnaires with parents. Setting: Two primary care practices in Oxfordshire, UK. Main outcome measures: The ability to collect capillary blood to test IAbs in children at the routine preschool vaccination (3.5–4 years). Results: Of 134 parents invited, 66 (49%) were recruited (median age 3.5 years (IQR 3.4–3.6), 26 (39.4%) male); 63 provided a sample (97% successfully), and one participant was identified with a single positive IAb. Parents (n=15 interviews, n=29 postcards) were uniformly positive about screening aligned to vaccination and stated they would have been less likely to take part had screening been a separate visit. Themes identified included preparedness for T1D and the long-term benefit outweighing short-term upset. The perceived volume of the capillary sample was a potential concern and needs optimising. Conclusions: Capillary IAb testing is a possible method to screen children for T1D. Aligning collection to the preschool vaccination visit can be convenient for families without the need for an additional visit

General population screening for type 1 diabetes using islet autoantibodies at the preschool vaccination visit: a proof-of-concept study (the T1Early study)

Objective: Type 1 diabetes (T1D) screening programmes testing islet autoantibodies (IAbs) in childhood can reduce life-threatening diabetic ketoacidosis. General population screening is required to detect the majority of children with T1D, since in >85% there is no family history. Age 3–5 years has been proposed as an optimal age for a single screen approach. Design: Capillary samples were collected from children attending their preschool vaccination and analysed for IAbs to insulin, glutamic acid decarboxylase, islet antigen-2 and zinc transporter 8 using radiobinding/luciferase immunoprecipitation system assays. Acceptability was assessed using semistructured interviews and open-ended postcard questionnaires with parents. Setting: Two primary care practices in Oxfordshire, UK. Main outcome measures: The ability to collect capillary blood to test IAbs in children at the routine preschool vaccination (3.5–4 years). Results: Of 134 parents invited, 66 (49%) were recruited (median age 3.5 years (IQR 3.4–3.6), 26 (39.4%) male); 63 provided a sample (97% successfully), and one participant was identified with a single positive IAb. Parents (n=15 interviews, n=29 postcards) were uniformly positive about screening aligned to vaccination and stated they would have been less likely to take part had screening been a separate visit. Themes identified included preparedness for T1D and the long-term benefit outweighing short-term upset. The perceived volume of the capillary sample was a potential concern and needs optimising. Conclusions: Capillary IAb testing is a possible method to screen children for T1D. Aligning collection to the preschool vaccination visit can be convenient for families without the need for an additional visit

Solar ultraviolet-B radiation and vitamin D: a cross-sectional population-based study using data from the 2007 to 2009 Canadian Health Measures Survey

Background: Exposure to solar ultraviolet-B (UV-B) radiation is a major source of vitamin D3. Chemistry climate models project decreases in ground-level solar erythemal UV over the current century. It is unclear what impact this will have on vitamin D status at the population level. The purpose of this study was to measure the association between ground-level solar UV-B and serum concentrations of 25-hydroxyvitamin D (25(OH)D) using a secondary analysis of the 2007 to 2009 Canadian Health Measures Survey (CHMS). Methods: Blood samples collected from individuals aged 12 to 79 years sampled across Canada were analyzed for 25(OH)D (n=4,398). Solar UV-B irradiance was calculated for the 15 CHMS collection sites using the Tropospheric Ultraviolet and Visible Radiation Model. Multivariable linear regression was used to evaluate the association between 25(OH)D and solar UV-B adjusted for other predictors and to explore effect modification. Results: Cumulative solar UV-B irradiance averaged over 91 days (91-day UV-B) prior to blood draw correlated significantly with 25(OH)D. Independent of other predictors, a 1 kJ/m 2 increase in 91-day UV-B was associated with a significant 0.5 nmol/L (95% CI 0.3-0.8) increase in mean 25(OH)D (P =0.0001). The relationship was stronger among younger individuals and those spending more time outdoors. Based on current projections of decreases in ground-level solar UV-B, we predict less than a 1 nmol/L decrease in mean 25(OH)D for the population. Conclusions: In Canada, cumulative exposure to ambient solar UV-B has a small but significant association with 25(OH)D concentrations. Public health messages to improve vitamin D status should target safe sun exposure with sunscreen use, and also enhanced dietary and supplemental intake and maintenance of a healthy body weight

Intraneuronal Aβ immunoreactivity is not a predictor of brain amyloidosis-β or neurofibrillary degeneration

Amyloid β (Aβ) immunoreactivity in neurons was examined in brains of 32 control subjects, 31 people with Down syndrome, and 36 patients with sporadic Alzheimer’s disease to determine if intraneuronal Aβ immunoreactivity is an early manifestation of Alzheimer-type pathology leading to fibrillar plaque formation and/or neurofibrillary degeneration. The appearance of Aβ immunoreactivity in neurons in infants and stable neuron-type specific Aβ immunoreactivity in a majority of brain structures during late childhood, adulthood, and normal aging does not support this hypothesis. The absence or detection of only traces of reaction with antibodies against 4–13 aa and 8–17 aa of Aβ in neurons indicated that intraneuronal Aβ was mainly a product of α- and γ-secretases (Aβ(17–40/42)). The presence of N-terminally truncated Aβ(17–40) and Aβ(17–42) in the control brains was confirmed by Western blotting and the identity of Aβ(17–40) was confirmed by mass spectrometry. The prevalence of products of α- and γ -secretases in neurons and β- and γ-secretases in plaques argues against major contribution of Aβ-immunopositive material detected in neuronal soma to amyloid deposit in plaques. The strongest intraneuronal Aβ(17–42) immunoreactivity was observed in structures with low susceptibility to fibrillar Aβ deposition, neurofibrillary degeneration, and neuronal loss compared to areas more vulnerable to Alzheimer-type pathology. These observations indicate that the intraneuronal Aβ immunoreactivity detected in this study is not a predictor of brain amyloidosis or neurofibrillary degeneration. The constant level of Aβ immunoreactivity in structures free from neuronal pathology during essentially the entire life span suggests that intraneuronal amino-terminally truncated Aβ represents a product of normal neuronal metabolism

The Metagenomics and Metadesign of the Subways and Urban Biomes (MetaSUB) International Consortium inaugural meeting report

The Metagenomics and Metadesign of the Subways and Urban Biomes (MetaSUB) International Consortium is a novel, interdisciplinary initiative comprised of experts across many fields, including genomics, data analysis, engineering, public health, and architecture. The ultimate goal of the MetaSUB Consortium is to improve city utilization and planning through the detection, measurement, and design of metagenomics within urban environments. Although continual measures occur for temperature, air pressure, weather, and human activity, including longitudinal, cross-kingdom ecosystem dynamics can alter and improve the design of cities. The MetaSUB Consortium is aiding these efforts by developing and testing metagenomic methods and standards, including optimized methods for sample collection, DNA/RNA isolation, taxa characterization, and data visualization. The data produced by the consortium can aid city planners, public health officials, and architectural designers. In addition, the study will continue to lead to the discovery of new species, global maps of antimicrobial resistance (AMR) markers, and novel biosynthetic gene clusters (BGCs). Finally, we note that engineered metagenomic ecosystems can help enable more responsive, safer, and quantified cities