Mycolic acids are major components of the cell wall of Mycobacterium tuberculosis. Several studies indicate that functional groups in the acyl chain of mycolic acids are important for pathogenesis ...and persistence. There are at least three mycolic acid cyclopropane synthases (PcaA, CmaA1, and CmaA2) that are responsible for these site-specific modifications of mycolic acids. To derive information on the specificity and enzyme mechanism of the family of proteins, the crystal structures of CmaA1, CmaA2, and PcaA were solved to 2-, 2-, and 2.65-Å resolution, respectively. All three enzymes have a seven-stranded α/β fold similar to other methyltransferases with the location and interactions with the cofactor S-adenosyl-l-methionine conserved. The structures of the ternary complexes demonstrate the position of the mycolic acid substrate binding site. Close examination of the active site reveals electron density that we believe represents a bicarbonate ion. The structures support the hypothesis that these enzymes catalyze methyl transfer via a carbocation mechanism in which the bicarbonate ion acts as a general base. In addition, comparison of the enzyme structures reveals a possible mechanism for substrate specificity. These structures provide a foundation for rational-drug design, which may lead to the development of new inhibitors effective against persistent bacteria.
To identify genetic variants associated with diabetic retinopathy (DR), we performed a large multiethnic genome-wide association study. Discovery included eight European cohorts (
= 3,246) and seven ...African American cohorts (
= 2,611). We meta-analyzed across cohorts using inverse-variance weighting, with and without liability threshold modeling of glycemic control and duration of diabetes. Variants with a
value <1 × 10
were investigated in replication cohorts that included 18,545 European, 16,453 Asian, and 2,710 Hispanic subjects. After correction for multiple testing, the C allele of rs142293996 in an intron of nuclear VCP-like (
) was associated with DR in European discovery cohorts (
= 2.1 × 10
), but did not reach genome-wide significance after meta-analysis with replication cohorts. We applied the Disease Association Protein-Protein Link Evaluator (DAPPLE) to our discovery results to test for evidence of risk being spread across underlying molecular pathways. One protein-protein interaction network built from genes in regions associated with proliferative DR was found to have significant connectivity (
= 0.0009) and corroborated with gene set enrichment analyses. These findings suggest that genetic variation in
as well as variation within a protein-protein interaction network that includes genes implicated in inflammation, may influence risk for DR.
CFTR F508del (c.1521_1523delCTT, p.Phe508delPhe) is the most common pathogenic allele underlying cystic fibrosis (CF), and its frequency varies in a geographic cline across Europe. We hypothesized ...that genetic variation associated with this cline is overrepresented in a large cohort (N > 5,000) of persons with CF who underwent whole-genome sequencing and that this pattern could result in spurious associations between variants correlated with both the F508del genotype and CF-related outcomes. Using principal-component (PC) analyses, we showed that variation in the CFTR region disproportionately contributes to a PC explaining a relatively high proportion of genetic variance. Variation near CFTR was correlated with population structure among persons with CF, and this correlation was driven by a subset of the sample inferred to have European ancestry. We performed genome-wide association studies comparing persons with CF with one versus two copies of the F508del allele; this allowed us to identify genetic variation associated with the F508del allele and to determine that standard PC-adjustment strategies eliminated the significant association signals. Our results suggest that PC adjustment can adequately prevent spurious associations between genetic variants and CF-related traits and are therefore effective tools to control for population structure even when population structure is confounded with disease severity and a common pathogenic variant.
Background. Genome-wide association studies (GWAS) may yield insights into longevity. Methods. We performed a meta-analysis of GWAS in Caucasians from four prospective cohort studies: the Age, ...Gene/Environment Susceptibility-Reykjavik Study, the Cardiovascular Health Study, the Framingham Heart Study, and the Rotterdam Study participating in the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) Consortium. Longevity was defined as survival to age 90 years or older (n = 1,836); the comparison group comprised cohort members who died between the ages of 55 and 80 years (n = 1,955). In a second discovery stage, additional genotyping was conducted in the Leiden Longevity Study cohort and the Danish 1905 cohort. Results. There were 273 single-nucleotide polymorphism (SNP) associations with p < .0001, but none reached the prespecified significance level of 5 × 10−8. Of the most significant SNPs, 24 were independent signals, and 16 of these SNPs were successfully genotyped in the second discovery stage, with one association for rs9664222, reaching 6.77 × 10−7 for the combined meta-analysis of CHARGE and the stage 2 cohorts. The SNP lies in a region near MINPP1 (chromosome 10), a well-conserved gene involved in regulation of cellular proliferation. The minor allele was associated with lower odds of survival past age 90 (odds ratio = 0.82). Associations of interest in a homologue of the longevity assurance gene (LASS3) and PAPPA2 were not strengthened in the second stage. Conclusion. Survival studies of larger size or more extreme or specific phenotypes may support or refine these initial findings.
The incidence of elbow medial ulnar collateral ligament (MUCL) injuries has been increasing, leading to advances in surgical treatments. However, it is not clear that there is consensus among ...surgeons regarding diagnostic imaging, the indications for acute surgery and postoperative rehabilitation. The purpose of this study is evaluate surgeon variability in the presurgical, surgical, and postsurgical treatment of MUCL injuries regarding the imaging modalities used for diagnosis, indications for acute surgical treatment, and postoperative treatment recommendations for rehabilitation and return to play (RTP). Our hypothesis is that indications for acute surgical treatment will be highly variable based on MUCL tear patterns and that agreement on the time to RTP will be consistent for throwing athletes and inconsistent for nonthrowing athletes.
A survey developed by 6 orthopedic surgeons with expertise in throwing athlete elbow injuries was distributed to 31 orthopedic surgeons who routinely treat MUCL injuries. The survey evaluated diagnostic and treatment topics related to MUCL injuries, and responses reaching 75% agreement were considered as high-level agreement.
Twenty-four surgeons responded to the survey, resulting in a 77% response rate. There is 75% or better agreement among surveyed surgeons regarding acute surgical treatment for distal full thickness tears, ulnar nerve transposition in symptomatic patients or with ulnar nerve subluxation, postoperative splinting for 1-2 weeks with initiation of rehabilitation within 2 weeks, the use of bracing after surgery and the initiation of a throwing program at 3 months after MUCL repair with internal brace by surgeons performing 20 or more MUCL surgeries per year. There were a considerable number of survey topics without high-level agreement, particularly regarding the indications for acute surgical treatment, the time to return to throwing and time RTP in both throwing and nonthrowing athletes.
The study reveals that there is agreement for the indication of acute surgical treatment of distal MUCL tears, duration of bracing after surgery, and the time to initiate physical therapy after surgery. There is not clear agreement on indications for surgical treatment for every MUCL tear pattern, RTP time for throwing, hitting and participation in nonthrowing sports.
Building stakeholders cannot easily quantify the environmental impacts of buildings as they accrue during construction. The goal of this work is to demonstrate a method to measure and manage the ...cradle-to-gate life cycle environmental impacts by linking environmental targets with modern construction management methods, to enable buildings to meet sustainable target values (STV). In this work, a construction activity-based computational framework was developed to enable stakeholders to reliably and efficiently construct cradle-to-gate life cycle models capturing environmental impacts including carbon and energy associated with material extraction, manufacture, transport to site, and construction. These models allow stakeholders to measure and manage impact accrual so as to not exceed STVs; without this framework, construction managers and other building stakeholders do not possess adequate environmental management tools to deliver projects consistently at or below STV. Specifically, the components developed are: (1) time dependent impact accrual budgets during construction and (2) impact measurement during construction. These benchmarks are used to determine whether a specific project is above or below target values, similar to methods for cost and schedule variance analysis. Two case studies were used to test this framework. This integration provides a life cycle assessment (LCA) modeling platform for management of environmental footprint during construction.
•Sustainable target values (STV) link building impacts with ecologic carrying capacity.•STV design connects LCA with building design, production, and construction.•STV is operationalized by integrating LCA and construction management practices.•A construction activity-based environmental management framework is developed.•With the framework, buildings can be produced that deliver on STV design.
Buildings are the largest consumer of energy and greatest contributor to climate change in the United States—consuming approximately half of energy produced and contributing close to half of ...greenhouse gas emissions. Building designers, contractors, and owners currently have few methods to effectively control a building's life cycle energy and environmental impacts during the design phase. Managing and reducing these impacts during design requires rapid information turnaround and decision-making. When left unconsidered, poor environmental design decisions leave potential design value uncaptured. This research combines life cycle assessment (LCA) and target value design (TVD) to rapidly produce more sustainable building designs. By establishing site-specific sustainability targets and using dynamically-updating life cycle assessments, this research demonstrates that buildings can be designed to perform at higher environmental standards than those designed without a target in place. The research also offers unique opportunities to analyze the tradeoffs between design and operational decisions.
•Sustainable target value (STV) design combines LCA and target value design (TVD).•Building life cycle performance targets are developed for CO2, energy, H2O, and O3.•STV targets are based on the planet's ecological carrying capacity.•STV software tool is developed to assess building design environmental performance.•Design teams with STV designed more sustainable buildings than teams without.
The impact of single-sport specialization on performance and injury risk in youth and adolescent athletes remains debated. The purpose of this study was to determine the incidence of single-sport ...specialization and its association with pitch velocity, pitcher utilization, range of motion, strength, and injury history.
High school pitchers in the Northeastern, Southeastern, Midwestern, and Western Regions of the United States were prospectively recruited. Prior to the season, pitchers were surveyed about their basic demographics, sport participation history, and injury history. At this timepoint, a standardized physical examination was performed and max pitch velocity was measured. They were then prospectively followed over the ensuing high school season, where pitch utilization and counts were followed and injuries were tracked.
Across the study, 115 pitchers were recruited, of whom 53% played baseball as their only sport and 47% played other sports. Single- position specialization within baseball was uncommon as 90% of athletes played positions other than pitcher. There were no differences in range of motion, strength, or pitch velocity between multisport athletes and single-sport specialists. Within the multisport group, 26% reported a history of injury, while 38% of the single-sport specialists reported a history of injury (P = .179). There were no significant differences in pitcher utilization between multisport athletes and single-sport specialists. Only a single prospective injury occurred in the 115 players.
Although single-sport specialization is common, nearly half of all high school pitchers in this study were multisport athletes. While injury history was generally similar between the two groups, single-sport baseball players did not have the following: improved motion, increased strength, higher ball velocity, or greater pitching volumes during the high school baseball season compared to multisport athletes. Accordingly, single-sport specialization did not have a competitive advantage for these high school pitchers based on the performance factors studied. Further prospective studies are necessary to understand the association between single-sport specialization and injury risk in high school baseball players.
Formation of globular clusters (GCs), the Galactic bulge, or galaxy bulges in general is an important unsolved problem in Galactic astronomy. Homogeneous infrared observations of large samples of ...stars belonging to GCs and the Galactic bulge field are one of the best ways to study these problems. We report the discovery by APOGEE (Apache Point Observatory Galactic Evolution Experiment) of a population of field stars in the inner Galaxy with abundances of N, C, and Al that are typically found in GC stars. The newly discovered stars have high N/Fe, which is correlated with Al/Fe and anticorrelated with C/Fe. They are homogeneously distributed across, and kinematically indistinguishable from, other field stars within the same volume. Their metallicity distribution is seemingly unimodal, peaking at Fe/H ~ -1, thus being in disagreement with that of the Galactic GC system. Our results can be understood in terms of different scenarios. N-rich stars could be former members of dissolved GCs, in which case the mass in destroyed GCs exceeds that of the surviving GC system by a factor of ~8. In that scenario, the total mass contained in so-called 'first-generation' stars cannot be larger than that in 'second-generation' stars by more than a factor of ~9 and was certainly smaller. Conversely, our results may imply the absence of a mandatory genetic link between 'second-generation' stars and GCs. Last, but not least, N-rich stars could be the oldest stars in the Galaxy, the by-products of chemical enrichment by the first stellar generations formed in the heart of the Galaxy.
Insulinomas are a rare cause of hyperinsulinaemic hypoglycaemia (HH) in children. The clinical features, investigations, management and histology of these rare pancreatic tumours in children have not ...been described in a large cohort of patients.
We conducted a retrospective review of cases diagnosed between 2000 and 2012, presenting to two referral centres in the United Kingdom. Clinical, biochemical, imaging (magnetic resonance imaging (MRI) and 6-L-¹⁸F-fluorodihydroxyphenylalanine (¹⁸F-DOPA) PET/CT scanning) and histological data were collected.
Nine children (age range 2-14.5 years) were diagnosed during the study period at Great Ormond Street Hospital (n=5) and Royal Manchester Children's Hospital (n=4). The combination of abdominal MRI scan (7/8) and ¹⁸F-DOPA PET/CT scan (2/4) correctly localised the anatomical location of all insulinomas. Before surgery, diazoxide therapy was used to treat hypoglycaemia, but only four patients responded. After surgical resection of the insulinoma, hypoglycaemia resolved in all patients. The anatomical localisation of the insulinoma in each patient was head (n=4), uncinate process (n=4) and tail (n=2, one second lesion) of the pancreas. Histology confirmed the diagnosis of insulinoma with the presence of sheets and trabeculae of epithelioid and spindle cells staining strongly for insulin and proinsulin, but not for glucagon or somatostatin. Two children were positive for MEN1, one of whom had two separate insulinoma lesions within the pancreas.
We describe a cohort of paediatric insulinoma patients. Although rare, insulinomas should be included in the differential diagnosis of HH, even in very young children. In the absence of a single imaging modality in the preoperative period, localisation of the tumour is achieved by combining imaging techniques, both conventional and functional.
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