Insight into the Carboxyl Transferase Domain Mechanism of Pyruvate Carboxylase from \u3cem\u3eRhizobium etli\u3c/em\u3e

The effects of mutations in the active site of the carboxyl transferase domain of Rhizobium etli pyruvate carboxylase have been determined for the forward reaction to form oxaloacetate, the reverse reaction to form MgATP, the oxamate-induced decarboxylation of oxaloacetate, the phosphorylation of MgADP by carbamoyl phosphate, and the bicarbonate-dependent ATPase reaction. Additional studies with these mutants examined the effect of pyruvate and oxamate on the reactions of the biotin carboxylase domain. From these mutagenic studies, putative roles for catalytically relevant active site residues were assigned and a more accurate description of the mechanism of the carboxyl transferase domain is presented. The T882A mutant showed no catalytic activity for reactions involving the carboxyl transferase domain but surprisingly showed 7- and 3.5-fold increases in activity, as compared to that of the wild-type enzyme, for the ADP phosphorylation and bicarbonate-dependent ATPase reactions, respectively. Furthermore, the partial inhibition of the T882A-catalyzed BC domain reactions by oxamate and pyruvate further supports the critical role of Thr882 in the proton transfer between biotin and pyruvate in the carboxyl transferase domain. The catalytic mechanism appears to involve the decarboxylation of carboxybiotin and removal of a proton from Thr882 by the resulting biotin enolate with either a concerted or subsequent transfer of a proton from pyruvate to Thr882. The resulting enolpyruvate then reacts with CO2 to form oxaloacetate and complete the reaction

Collective Action and Football Fandom A Relational Sociological Approach

This book draws upon a relational sociological paradigm to explore the processes of collective action in football fandom across Europe and the UK. Through a range of case studies, the authors address pertinent themes in football fandom, ..

Comparing CNN and Human Crafted Features for Human Activity Recognition

Deep learning techniques such as Convolutional Neural Networks (CNNs) have shown good results in activity recognition. One of the advantages of using these methods resides in their ability to generate features automatically. This ability greatly simplifies the task of feature extraction that usually requires domain specific knowledge, especially when using big data where data driven approaches can lead to anti-patterns. Despite the advantage of this approach, very little work has been undertaken on analyzing the quality of extracted features, and more specifically on how model architecture and parameters affect the ability of those features to separate activity classes in the final feature space. This work focuses on identifying the optimal parameters for recognition of simple activities applying this approach on both signals from inertial and audio sensors. The paper provides the following contributions: (i) a comparison of automatically extracted CNN features with gold standard Human Crafted Features (HCF) is given, (ii) a comprehensive analysis on how architecture and model parameters affect separation of target classes in the feature space. Results are evaluated using publicly available datasets. In particular, we achieved a 93.38% F-Score on the UCI-HAR dataset, using 1D CNNs with 3 convolutional layers and 32 kernel size, and a 90.5% F-Score on the DCASE 2017 development dataset, simplified for three classes (indoor, outdoor and vehicle), using 2D CNNs with 2 convolutional layers and a 2x2 kernel size

Probing the Catalytic Roles of Arg548 and Gln552 in the Carboxyl Transferase Domain of the \u3cem\u3eRhizobium etli\u3c/em\u3e Pyruvate Carboxylase by Site-directed Mutagenesis

The roles of Arg548 and Gln552 residues in the active site of the carboxyl transferase domain of Rhizobium etli pyruvate carboxylase were investigated using site-directed mutagenesis. Mutation of Arg548 to alanine or glutamine resulted in the destabilization of the quaternary structure of the enzyme, suggesting that this residue has a structural role. Mutations R548K, Q552N, and Q552A resulted in a loss of the ability to catalyze pyruvate carboxylation, biotin-dependent decarboxylation of oxaloacetate, and the exchange of protons between pyruvate and water. These mutants retained the ability to catalyze reactions that occur at the active site of the biotin carboxylase domain, i.e., bicarbonate-dependent ATP cleavage and ADP phosphorylation by carbamoyl phosphate. The effects of oxamate on the catalysis in the biotin carboxylase domain by the R548K and Q552N mutants were similar to those on the catalysis of reactions by the wild-type enzyme. However, the presence of oxamate had no effect on the reactions catalyzed by the Q552A mutant. We propose that Arg548 and Gln552 facilitate the binding of pyruvate and the subsequent transfer of protons between pyruvate and biotin in the partial reaction catalyzed in the active site of the carboxyl transferase domain of Rhizobium etli pyruvate carboxylase

Influence of case definition on incidence and outcome of acute coronary syndromes

© 2016, BMJ Publishing Group. All rights reserved. Objective: Acute coronary syndromes (ACS) are common, but their incidence and outcome might depend greatly on how data are collected. We compared case ascertainment rates for ACS and myocardial infarction (MI) in a single institution using several different strategies. Methods: The Hull and East Yorkshire Hospitals serve a population of ∼560 000. Patients admitted with ACS to cardiology or general medical wards were identified prospectively by trained nurses during 2005. Patients with a death or discharge code of MI were also identified by the hospital information department and, independently, from Myocardial Infarction National Audit Project (MINAP) records. The hospital laboratory identified all patients with an elevated serum troponin-T (TnT) by contemporary criteria ( > 0.03 μg/L in 2005). Results: The prospective survey identified 1731 admissions (1439 patients) with ACS, including 764 admissions (704 patients) with MIs. The hospital information department reported only 552 admissions (544 patients) with MI and only 206 admissions (203 patients) were reported to the MINAP. Using all 3 strategies, 934 admissions (873 patients) for MI were identified, for which TnT was > 1 μg/L in 443, 0.04-1.0 μg/L in 435, =0.03 μg/L in 19 and not recorded in 37. A further 823 patients had TnT > 0.03 μg/L, but did not have ACS ascertained by any survey method. Of the 873 patients with MI, 146 (16.7%) died during admission and 218 (25.0%) by 1 year, but ranging from 9% for patients enrolled in the MINAP to 27% for those identified by the hospital information department. Conclusions: MINAP and hospital statistics grossly underestimated the incidence of MI managed by our hospital. The 1-year mortality was highly dependent on the method of ascertainment

Collegiate Athletes’ Perceptions of Osteopathic Manipulative Treatment

Introduction. Many published studies have examined the effects of osteopathic manipulative treatment (OMT), but none has evaluated its role in treating collegiate athletes. The authors examined collegiate athletes’ perception of OMT. Methods. A cross-sectional survey of a convenience sample of 592 collegiate athletes was conducted from two universities in the midwestern United States during August-September 2019. The athletes completed a 12-item survey during pre-participation physical evaluations at their respective institutions. Main outcome measures included pain, need for pain medication, stress and anxiety associated with injuries, and overall satisfaction with the OMT in recovery and return to sports. Fisher’s exact test was used to evaluate association between the variables. Results. The participation rate was 80.6% (477/592). Slightly fewer than 7% (31/477) of the athletes were familiar with OMT. Eighteen of the 31 athletes (58.1%) had received osteopathic manipulation as part of a treatment plan for injury. Of these athletes, 94.4% (17/18) reported a decreased need for pain medication and 83.3% (15/18) had reduced stress and anxiety related to their injury. One in three of them expressed interest in receiving osteopathic manipulation as a treatment option for an injury. The athletes reported general satisfaction with OMT in their recovery and return to sports. Conclusion. The findings demonstrated the interest and benefits of OMT among collegiate athletes. This evidence supported previous findings about perceived efficacy of OMT in treating patients regardless of injury and diagnosis. Future studies need to establish causal relationship among OMT, stress and anxiety, pain, and use of pain medications

Academic Impact of COVID-19 in Collegiate Athletes

Introduction. The COVID-19 pandemic caused a pause to nearly all sporting activities in the spring of 2020, and collegiate athletes at the National Collegiate Athletic Association (NCAA)-affiliated universities whose sporting seasons were affected by the pandemic were granted an extra year of athletic eligibility. The study was conducted to determine how collegiate athletes planned to use an additional year of eligibility granted by the NCAA. Methods. The authors conducted a cross-sectional survey of 632 athletes from two universities in the Midwestern United States, between August and September 2021. The athletes completed an anonymous, 9-item survey to assess the effect of the pandemic on athletic season, athletic eligibility, and potential change in an academic or professional career. Chi-square tests, generalized linear mixed models, and adjusted odds ratio were used for the analyses. Results. The participation rate was 74.5% (471 of 632). Nearly 63% (290 of 461) of the athletes received an additional year of eligibility because of the pandemic, with 193 (66.6%) planned to use their extra year for scholastic development. Male athletes (65.3% vs 34.7%; χ2[1, n=290]=11.66, P<.001, Φ=0.20), Division II athletes (59.6% vs 40.4%; χ2[1, n=290]=13.93, P<.001, Φ=0.22), and athletes who had not previously used redshirt (73.1% vs 26.9%; χ2[1, n=290] = 4.79, P=.029, Φ=0.32) where more likely to use their extra year of eligibility academically. Conclusions. Our findings suggest that most of the athletes planned to use their extra year of eligibility to pursue further scholastic or professional development, highlighting the positive part of the COVID-19 pandemic. Future studies should investigate how these findings relate to athletes from universities in different geographical locations, intra-division schools

Diabetic retinopathy in Tanzania: prevalence and risk factors at entry into a regional screening programme.

OBJECTIVE: The number of adults with diabetes in sub-Saharan Africa (SSA) is expected to almost double by 2035. This study investigated the prevalence of diabetic retinopathy (DR) and its risk factors at entry into a community-based screening programme. METHODS: All persons with diabetes screened for retinopathy at entry into a screening programme in Kilimanjaro Region, Tanzania between November 2010 and December 2014 were included. Fundus photographs were taken with a Topcon retinal camera following pupil dilation. Data were collected on BP, random blood sugar, duration of diabetes, BMI and visual acuity on entry. RESULTS: A total of 3187 persons were screened for DR. The prevalence of any DR was 27.9% (95%CI 26.4-29.5%) with background diabetic retinopathy (BDR), pre-proliferative diabetic retinopathy (PPDR) and proliferative diabetic retinopathy (PDR) having a prevalence of 19.1% (95% CI 17.7-20.4%), 6.0% (95%CI 5.2-6.8%) and 2.9% (95%CI 2.3-3.5%), respectively. Maculopathy was present in 16.1% (95%CI 14.8-17.4%) of participants. Multivariable logistic regression analysis for the presence of any DR found independent associations with duration of diabetes (P < 0.0001), systolic BP (P < 0.0001), random blood sugar (P < 0.0001) and attending a government hospital diabetic clinic (P = 0.0339). CONCLUSIONS: This study is the first to present data from a DR screening programme in SSA. The results will provide policymakers with data to aid planning of DR screening and treatment services in the African region. The study highlights the importance of managing comorbidities within DR screening programmes