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1.
Magn Reson Med ; 91(4): 1586-1597, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38169132

RESUMO

PURPOSE: To develop a tissue field-filtering algorithm, called maximum spherical mean value (mSMV), for reducing shadow artifacts in QSM of the brain without requiring brain-tissue erosion. THEORY AND METHODS: Residual background field is a major source of shadow artifacts in QSM. The mSMV algorithm filters large field-magnitude values near the border, where the maximum value of the harmonic background field is located. The effectiveness of mSMV for artifact removal was evaluated by comparing existing QSM algorithms in numerical brain simulation as well as using in vivo human data acquired from 11 healthy volunteers and 93 patients. RESULTS: Numerical simulation showed that mSMV reduces shadow artifacts and improves QSM accuracy. Better shadow reduction, as demonstrated by lower QSM variation in the gray matter and higher QSM image quality score, was also observed in healthy subjects and in patients with hemorrhages, stroke, and multiple sclerosis. CONCLUSION: The mSMV algorithm allows QSM maps that are substantially equivalent to those obtained using SMV-filtered dipole inversion without eroding the volume of interest.


Assuntos
Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Humanos , Processamento de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Imagens de Fantasmas , Encéfalo/diagnóstico por imagem , Mapeamento Encefálico/métodos , Algoritmos , Artefatos
2.
J Magn Reson Imaging ; 57(6): 1621-1640, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36748806

RESUMO

Magnetic materials in tissue, such as iron, calcium, or collagen, can be studied using quantitative susceptibility mapping (QSM). To date, QSM has been overwhelmingly applied in the brain, but is increasingly utilized outside the brain. QSM relies on the effect of tissue magnetic susceptibility sources on the MR signal phase obtained with gradient echo sequence. However, in the body, the chemical shift of fat present within the region of interest contributes to the MR signal phase as well. Therefore, correcting for the chemical shift effect by means of water-fat separation is essential for body QSM. By employing techniques to compensate for cardiac and respiratory motion artifacts, body QSM has been applied to study liver iron and fibrosis, heart chamber blood and placenta oxygenation, myocardial hemorrhage, atherosclerotic plaque, cartilage, bone, prostate, breast calcification, and kidney stone.


Assuntos
Algoritmos , Imageamento por Ressonância Magnética , Masculino , Humanos , Imageamento por Ressonância Magnética/métodos , Fígado , Ferro , Abdome , Encéfalo , Mapeamento Encefálico
3.
J Sports Sci ; 41(8): 788-795, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37537860

RESUMO

Coaches use physically constraining tools to supplement their coaching when developing sport-specific skills, however, their effectiveness is unknown. This scoping review aimed to understand the efficacy of physically constraining tools used in racket, stick, and club sports for grip-specific skill development. This scoping review followed the Joanna Briggs Institute methodology and PRISMA guidelines. Peer-reviewed research, including quasi-experimental, true-experimental, case studies, and grey literature were considered. Peer-reviewed sources were searched on Web of Science, Medline, and SPORTDiscus until October 6th, 2022. Exclusion criteria were (1) not original peer-reviewed research; (2) disabled participants or used for rehabilitation; or (3) not available in English. Data extracted were the type of tool, research foci, measures, and outcomes of the tool's efficacy. Zero peer-reviewed sources were identified on the efficacy of using physically constraining tools for grip-specific skill development. Common trends identified from the excluded sources were explored to provide a basis for the importance of using physically constraining tools for grip-specific skill development. Many tools are used in coaching despite their unclear efficacy, however, the current results can guide future work to assess the acute and longitudinal effects of using these tools, specifically within the development and performance of sport-specific skills.


Assuntos
Esportes , Humanos , Suplementos Nutricionais
4.
J Magn Reson Imaging ; 56(2): 450-461, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35343008

RESUMO

BACKGROUND: Methods for accurate quantification of lung fluid in heart failure (HF) are needed. Dynamic contrast-enhanced (DCE)-MRI may be an appropriate modality. PURPOSE: DCE-MRI evaluation of fraction of fluid volume in the interstitial lung space (ve ) and vascular permeability (Ktrans ). STUDY TYPE: Prospective, single-center method validation. POPULATION: Seventeen evaluable healthy volunteers (HVs), 12 participants with HF, and 3 with acute decompensated HF (ADHF). FIELD STRENGTH/SEQUENCE: T1 mapping (spoiled gradient echo variable flip angle acquisition) followed by dynamic series (three-dimensional spoiled gradient-recalled echo acquisitions [constant echo time, repetition time, and flip angle at 1.5 T]). ASSESSMENT: Three whole-chest scans were acquired: baseline (Session 1), 1-week later (Session 2), following exercise (Session 3). Extended Tofts model quantified ve and Ktrans (voxel-wise basis); total lung median measures were extracted and fitted via repeat measure analysis of variance (ANOVA) model. Patient tolerability of the scanning protocol was assessed. STATISTICAL TESTS: This was constructed as an experimental medicine study. PRIMARY ENDPOINTS: Ktrans and ve at baseline (HV vs. HF), change in Ktrans and ve following exercise, and following lung congestion resolution (ADHF). Ktrans and ve were fitted separately using ANOVA. Secondary endpoint: repeatability, that is, within-participant variability in ve and Ktrans between sessions (coefficient of variation estimated via mixed effects model). RESULTS: There was no significant difference in mean Ktrans between HF and HV (P ≤ 0.17): 0.2216 minutes-1 and 0.2353 minutes-1 (Session 1), 0.2044 minutes-1 and 0.2567 minutes-1 (Session 2), 0.1841 minutes-1 and 0.2108 minutes-1 (Session 3), respectively. ve was greater in the HF group (all scans, P ≤ 0.02). Results were repeatable between Sessions 1 and 2; mean values for HF and HV were 0.4946 and 0.3346 (Session 1), 0.4353 and 0.3205 (Session 2), respectively. There was minimal difference in Ktrans or ve between scans for participants with ADHF (small population precluded significance testing). Scanning was well tolerated. DATA CONCLUSION: While no differences were detected in Ktrans , ve was greater in chronic HF patients vs. HV, augmented beyond plasma and intracellular volume. DCE-MRI is a valuable diagnostic and physiologic tool to evaluate changes in fluid volume in the interstitial lung space associated with symptomatic HF. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY STAGE: 2.


Assuntos
Meios de Contraste , Insuficiência Cardíaca , Insuficiência Cardíaca/diagnóstico por imagem , Humanos , Pulmão/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Permeabilidade
5.
J Strength Cond Res ; 36(2): 493-497, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31895279

RESUMO

ABSTRACT: Roberts, AH, Walden, AJ, Carter, KA, and Symons, TB. Effect of sport-specific constraints on aerobic capacity in high school field hockey players. J Strength Cond Res 36(2): 493-497, 2022-Many sports require the use of a mouth guard (MG) and carrying an object during play. Research has shown that individually these conditions can cause performance decrements; however, no research has been conducted into the effect of combining both mouth guard and stick with adolescent female field hockey players. This study aimed to determine the effects of MG and stick (STK) use on estimated aerobic capacity over the course of a season long training and competition period. Thirty-eight female high school field hockey players were separated into 2 groups: experimental (EXP-all training with MG-STK) or control (CTL-all training without MG-STK). Aerobic capacity was estimated using a multi-stage fitness test. Field hockey specific training prescribed by coaching staff was performed throughout the study, with testing at pre-, mid- and post-season. Subjects performed 2 sets of testing at each time point, first without MG-STK (WOMG-STK) and then with MG-STK. No main effect was observed in aerobic capacity estimations between groups; however, MG-STK testing reduced estimated aerobic capacity at each time point, regardless of the group (WOMG-STK: 37.4 ± 6. mL·kg-1·min-1 vs. MG-STK: 33.2 ± 4. mL·kg-1·min-1; p < 0.01). Chronic use of MG-STK does not seem to negatively impact estimates of aerobic capacity over the course of a season in high school field hockey players. Therefore, players should be encouraged to use MGs during all training sessions as it increases safety and familiarity with no decrement in long-term physiological performance.


Assuntos
Hóquei , Adolescente , Exercício Físico , Teste de Esforço , Tolerância ao Exercício , Feminino , Humanos , Instituições Acadêmicas
6.
Eur Radiol ; 31(8): 5746-5758, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33591383

RESUMO

OBJECTIVES: Evaluate test-retest repeatability, ability to discriminate between osteoarthritic and healthy participants, and sensitivity to change over 6 months, of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) biomarkers in knee OA. METHODS: Fourteen individuals aged 40-60 with mild-moderate knee OA and 6 age-matched healthy volunteers (HV) underwent DCE-MRI at 3 T at baseline, 1 month and 6 months. Voxelwise pharmacokinetic modelling of dynamic data was used to calculate DCE-MRI biomarkers including Ktrans and IAUC60. Median DCE-MRI biomarker values were extracted for each participant at each study visit. Synovial segmentation was performed using both manual and semiautomatic methods with calculation of an additional biomarker, the volume of enhancing pannus (VEP). Test-retest repeatability was assessed using intraclass correlation coefficients (ICC). Smallest detectable differences (SDDs) were calculated from test-retest data. Discrimination between OA and HV was assessed via calculation of between-group standardised mean differences (SMD). Responsiveness was assessed via the number of OA participants with changes greater than the SDD at 6 months. RESULTS: Ktrans demonstrated the best test-retest repeatability (Ktrans/IAUC60/VEP ICCs 0.90/0.84/0.40, SDDs as % of OA mean 33/71/76%), discrimination between OA and HV (SMDs 0.94/0.54/0.50) and responsiveness (5/1/1 out of 12 OA participants with 6-month change > SDD) when compared to IAUC60 and VEP. Biomarkers derived from semiautomatic segmentation outperformed those derived from manual segmentation across all domains. CONCLUSIONS: Ktrans demonstrated the best repeatability, discrimination and sensitivity to change suggesting that it is the optimal DCE-MRI biomarker for use in experimental medicine studies. KEY POINTS: • Dynamic contrast-enhanced MRI (DCE-MRI) provides quantitative measures of synovitis in knee osteoarthritis which may permit early assessment of efficacy in experimental medicine studies. • This prospective observational study compared DCE-MRI biomarkers across domains relevant to experimental medicine: test-retest repeatability, discriminative validity and sensitivity to change. • The DCE-MRI biomarker Ktrans demonstrated the best performance across all three domains, suggesting that it is the optimal biomarker for use in future interventional studies.


Assuntos
Osteoartrite do Joelho , Sinovite , Meios de Contraste , Humanos , Lactente , Articulação do Joelho , Imageamento por Ressonância Magnética , Osteoartrite do Joelho/diagnóstico por imagem , Estudos Prospectivos , Sinovite/diagnóstico por imagem
7.
J Sports Sci ; 39(4): 359-367, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32962508

RESUMO

Coaches are an integral part of talent identification in sport and are often used as the "gold standard" against which scientific methods of talent identification are compared. However, their decision-making during this process is not well understood. In this article, we use an ecological approach to explore talent identification in combat sports. We interviewed twenty-four expert, international-level coaches from the Olympic disciplines of boxing, judo, and taekwondo (age: 48.7 + 7.5 years; experience: 20.8 + 8.3 years). Findings indicated that when coaches identify talent they rely on "gut instinct": intuitive judgements made without conscious thought, used to direct attention to particular athletes or characteristics. Our analysis revealed four major contributors to coaches' intuition: experiential knowledge, temporal factors, seeing athletes in context, and what can be worked with. Our findings demonstrate that i) athlete selections may be influenced by the coaches' perceived ability to improve certain athletes (rather than solely on athlete ability); and ii) "instinctual" decisions are the result of years of experience, time spent with the athlete, and the context surrounding the decision. Based on these findings, we recommend that future research focuses on the duration and conditions that are required for coaches to confidently and reliably identify talented athletes.


Assuntos
Aptidão , Atletas , Desempenho Atlético , Tomada de Decisões/fisiologia , Instinto , Intuição/fisiologia , Boxe , Feminino , Humanos , Julgamento/fisiologia , Masculino , Artes Marciais , Pessoa de Meia-Idade , Pesquisa Qualitativa , Fatores de Tempo
8.
J Magn Reson Imaging ; 52(6): 1753-1764, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32677070

RESUMO

BACKGROUND: Determining the compositional response of articular cartilage to dynamic joint-loading using MRI may be a more sensitive assessment of cartilage status than conventional static imaging. However, distinguishing the effects of joint-loading vs. inherent measurement variability remains difficult, as the repeatability of these quantitative methods is often not assessed or reported. PURPOSE: To assess exercise-induced changes in femoral, tibial, and patellar articular cartilage composition and compare these against measurement repeatability. STUDY TYPE: Prospective observational study. POPULATION: Phantom and 19 healthy participants. FIELD STRENGTH/SEQUENCE: 3T; 3D fat-saturated spoiled gradient recalled-echo; T1ρ - and T2 -prepared pseudosteady-state 3D fast spin echo. ASSESSMENT: The intrasessional repeatability of T1ρ and T2 relaxation mapping, with and without knee repositioning between two successive measurements, was determined in 10 knees. T1ρ and T2 relaxation mapping of nine knees was performed before and at multiple timepoints after a 5-minute repeated, joint-loading stepping activity. 3D surface models were created from patellar, femoral, and tibial articular cartilage. STATISTICAL TESTS: Repeatability was assessed using root-mean-squared-CV (RMS-CV). Using Bland-Altman analysis, thresholds defined as the smallest detectable difference (SDD) were determined from the repeatability data with knee repositioning. RESULTS: Without knee repositioning, both surface-averaged T1ρ and T2 were very repeatable on all cartilage surfaces, with RMS-CV <1.1%. Repositioning of the knee had the greatest effect on T1ρ of patellar cartilage with the surface-averaged RMS-CV = 4.8%. While T1ρ showed the greatest response to exercise at the patellofemoral cartilage region, the largest changes in T2 were determined in the lateral femorotibial region. Following thresholding, significant (>SDD) average exercise-induced in T1ρ and T2 of femoral (-8.0% and -5.3%), lateral tibial (-6.9% and -5.9%), medial tibial (+5.8% and +2.9%), and patellar (-7.9% and +2.8%) cartilage were observed. DATA CONCLUSION: Joint-loading with a stepping activity resulted in T1ρ and T2 changes above background measurement error. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY STAGE: 1 J. MAGN. RESON. IMAGING 2020;52:1753-1764.


Assuntos
Cartilagem Articular , Cartilagem Articular/diagnóstico por imagem , Humanos , Joelho , Articulação do Joelho/diagnóstico por imagem , Imageamento por Ressonância Magnética , Tíbia/diagnóstico por imagem
9.
J Magn Reson Imaging ; 52(4): 1139-1151, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32447815

RESUMO

BACKGROUND: Traditional quantitative analysis of cartilage with MRI averages measurements (eg, thickness) across regions-of-interest (ROIs) which may reduce responsiveness. PURPOSE: To validate and describe clinical application of a semiautomated surface-based method for analyzing cartilage relaxation times ("composition") and morphology on MRI, 3D cartilage surface mapping (3D-CaSM). STUDY TYPE: Validation study in cadaveric knees and prospective observational (cohort) study in human participants. POPULATION: Four cadaveric knees and 14 participants aged 40-60 with mild-moderate knee osteoarthritis (OA) and 6 age-matched healthy volunteers, imaged at baseline, 1, and 6 months. FIELD STRENGTH/SEQUENCE: 3D spoiled gradient echo, T1 rho/T2 magnetization-prepared 3D fast spin echo for mapping of T1 rho/T2 relaxation times and delayed gadolinium enhanced MRI of cartilage (dGEMRIC) using variable flip angle T1 relaxation time mapping at 3T. ASSESSMENT: 3D-CaSM was validated against high-resolution peripheral quantitative computed tomography (HRpQCT) in cadaveric knees, with comparison to expert manual segmentation. The clinical study assessed test-retest repeatability and sensitivity to change over 6 months for cartilage thickness and relaxation times. STATISTICAL TESTS: Bland-Altman analysis was performed for the validation study and evaluation of test-retest repeatability. Six-month changes were assessed via calculation of the percentage of each cartilage surface affected by areas of significant change (%SC), defined using thresholds based on area and smallest detectable difference (SDD). RESULTS: Bias and precision (0.06 ± 0.25 mm) of 3D-CaSM against reference HRpQCT data were comparable to expert manual segmentation (-0.13 ± 0.26 mm). 3D-CaSM demonstrated significant (>SDD) 6-month changes in cartilage thickness and relaxation times in both OA participants and healthy controls. The parameter demonstrating the greatest 6-month change was T2 relaxation time (OA median %SC [IQR] = 8.8% [5.5 to 12.6]). DATA CONCLUSION: This study demonstrates the construct validity and potential clinical utility of 3D-CaSM, which may offer advantages to conventional ROI-based methods. LEVEL OF EVIDENCE: 2. TECHNICAL EFFICACY STAGE: 2. J. Magn. Reson. Imaging 2020;52:1139-1151.


Assuntos
Cartilagem Articular , Osteoartrite do Joelho , Adulto , Cartilagem Articular/diagnóstico por imagem , Gadolínio DTPA , Humanos , Lactente , Articulação do Joelho , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Osteoartrite do Joelho/diagnóstico por imagem , Reprodutibilidade dos Testes
10.
Cell Microbiol ; 21(10): e13089, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31373151

RESUMO

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), is a skin disease responsible for the global decline of amphibians. Frog species and populations can vary in susceptibility, but this phenomenon remains poorly understood. Here, we investigated serotonin in the skin of infected and uninfected frogs. In more susceptible frog populations, skin serotonin rose with increasing infection intensity, but decreased in later stages of the disease. The more resistant population maintained a basal level of skin serotonin. Serotonin inhibited both Bd sporangial growth and Jurkat lymphocyte proliferation in vitro. However, serotonin accumulates in skin granular glands, and this compartmentalisation may prevent inhibition of Bd growth in vivo. We suggest that skin serotonin increases in susceptible frogs due to pathogen excretion of precursor tryptophan, but that resistant frogs are able to control the levels of serotonin. Overall, the immunosuppressive effects of serotonin may contribute to the susceptibility of frogs to chytridiomycosis.


Assuntos
Anuros/microbiologia , Quitridiomicetos , Suscetibilidade a Doenças/veterinária , Micoses/veterinária , Serotonina/metabolismo , Dermatopatias/veterinária , Pele/metabolismo , Animais , Anuros/imunologia , Anuros/metabolismo , Austrália , Proliferação de Células/efeitos dos fármacos , Quitridiomicetos/efeitos dos fármacos , Suscetibilidade a Doenças/metabolismo , Suscetibilidade a Doenças/microbiologia , Cromatografia Gasosa-Espectrometria de Massas , Micoses/imunologia , Micoses/metabolismo , Serotonina/farmacologia , Pele/química , Pele/microbiologia , Dermatopatias/metabolismo , Esporângios/efeitos dos fármacos , Esporângios/crescimento & desenvolvimento , Linfócitos T/efeitos dos fármacos
11.
Med Mycol ; 57(2): 204-214, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29566178

RESUMO

Captive and wild amphibians are under threat of extinction from the deadly fungal pathogen Batrachochytrium dendrobatidis (Bd). The antifungal drug terbinafine (TBF) is used by pet owners to treat Bd-infected frogs; however, it is not widely used in academic or zoological institutions due to limited veterinary clinical trials. To assess TBF's efficacy, we undertook treatment trials and pharmacokinetic studies to investigate drug absorption and persistence in frog skin; and then we correlated these data to the minimal lethal concentrations (MLC) against Bd. Despite an initial reduction in zoospore load, the recommended treatment (five daily 5 min 0.01% TBF baths) was unable to cure experimentally infected alpine tree frogs and naturally infected common eastern froglets. In vitro and in vivo pharmacokinetics showed that absorbed TBF accumulates in frog skin with increased exposure, indicating its suitability for treating cutaneous pathogens via direct application. The MLC of TBF for zoosporangia was 100 µg/ml for 2 h, while the minimal inhibitory concentration was 2 µg/ml, suggesting that the drug concentration absorbed during 5 min treatments is not sufficient to cure high Bd burdens. With longer treatments of five daily 30 min baths, Bd clearance improved from 12.5% to 50%. A higher dose of 0.02% TBF resulted in 78% of animals cured; however, clearance was not achieved in all individuals due to low TBF skin persistence, as the half-life was less than 2 h. Therefore, the current TBF regime is not recommended as a universal treatment against Bd until protocols are optimized, such as with increased exposure frequency.


Assuntos
Antifúngicos/administração & dosagem , Antifúngicos/farmacocinética , Anuros/microbiologia , Quitridiomicetos/efeitos dos fármacos , Micoses/veterinária , Terbinafina/administração & dosagem , Terbinafina/farmacocinética , Animais , Antifúngicos/farmacologia , Relação Dose-Resposta a Droga , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Micoses/tratamento farmacológico , Pele/efeitos dos fármacos , Pele/metabolismo , Pele/microbiologia , Esporos Fúngicos/efeitos dos fármacos , Terbinafina/farmacologia , Resultado do Tratamento
12.
Pediatr Exerc Sci ; 31(4): 448-457, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30982436

RESUMO

PURPOSE: A novel 4-task Athlete Introductory Movement Screen was developed and tested to provide an appropriate and reliable movement screening tool for youth sport practitioners. METHODS: The overhead squat, lunge, push-up, and a prone brace with shoulder touches were selected based on previous assessments. A total of 28 mixed-sport junior athletes (18 boys and 10 girls; mean age = 15.7 [1.8] y) completed screening after viewing standardized demonstration videos. Athletes were filmed performing 8 repetitions of each task and assessed retrospectively by 2 independent raters using a 3-point scale. The primary rater reassessed the footage 3 weeks later. A subgroup (n = 11) repeated the screening 7 days later, and a further 8 athletes were reassessed 6 months later. Intraclass correlation coefficients (ICC), typical error (TE), coefficient of variation (CV%), and weighted kappa (k) were used in reliability analysis. RESULTS: For the Athlete Introductory Movement Screen 4-task sum score, intrarater reliability was high (ICC = .97; CV = 2.8%), whereas interrater reliability was good (intraclass correlation coefficient = .88; CV = 5.6%). There was a range of agreement from fair to almost perfect (k = .31-.89) between raters across individual movements. A 7-day and 6-month test-retest held good reliability and acceptable CVs (≤ 10%) for sum scores. CONCLUSION: The 4-task Athlete Introductory Movement Screen appears to be a reliable tool for profiling emerging athletes. Reliability was strongest within the same rater; it was lower, yet acceptable, between 2 raters. Scores can provide an overview of appropriate movement competencies, helping practitioners assess training interventions in the athlete development pathway.


Assuntos
Atletas , Teste de Esforço/métodos , Movimento , Esportes Juvenis , Adolescente , Feminino , Humanos , Masculino , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Estudos Retrospectivos , Gravação em Vídeo
13.
Infect Immun ; 82(1): 316-32, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24166956

RESUMO

In Staphylococcus aureus, the low-molecular-weight thiol called bacillithiol (BSH), together with cognate S-transferases, is believed to be the counterpart to the glutathione system of other organisms. To explore the physiological role of BSH in S. aureus, we constructed mutants with the deletion of bshA (sa1291), which encodes the glycosyltransferase that catalyzes the first step of BSH biosynthesis, and fosB (sa2124), which encodes a BSH-S-transferase that confers fosfomycin resistance, in several S. aureus strains, including clinical isolates. Mutation of fosB or bshA caused a 16- to 60-fold reduction in fosfomycin resistance in these S. aureus strains. High-pressure liquid chromatography analysis, which quantified thiol extracts, revealed some variability in the amounts of BSH present across S. aureus strains. Deletion of fosB led to a decrease in BSH levels. The fosB and bshA mutants of strain COL and a USA300 isolate, upon further characterization, were found to be sensitive to H2O2 and exhibited decreased NADPH levels compared with those in the isogenic parents. Microarray analyses of COL and the isogenic bshA mutant revealed increased expression of genes involved in staphyloxanthin synthesis in the bshA mutant relative to that in COL under thiol stress conditions. However, the bshA mutant of COL demonstrated decreased survival compared to that of the parent in human whole-blood survival assays; likewise, the naturally BSH-deficient strain SH1000 survived less well than its BSH-producing isogenic counterpart. Thus, the survival of S. aureus under oxidative stress is facilitated by BSH, possibly via a FosB-mediated mechanism, independently of its capability to produce staphyloxanthin.


Assuntos
Proteínas de Bactérias/fisiologia , Cisteína/análogos & derivados , Glucosamina/análogos & derivados , Estresse Oxidativo/fisiologia , Staphylococcus aureus/metabolismo , Amidoidrolases/deficiência , Análise de Variância , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Cisteína/genética , Cisteína/fisiologia , Glucosamina/genética , Glucosamina/fisiologia , Glicosiltransferases/genética , Peróxido de Hidrogênio/farmacologia , Análise em Microsséries , Testes de Sensibilidade Microbiana , Mutação , NADP/metabolismo , Peroxidase/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/crescimento & desenvolvimento , Xantofilas/biossíntese
14.
Biochem J ; 451(1): 69-79, 2013 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-23256780

RESUMO

FosB is a divalent-metal-dependent thiol-S-transferase implicated in fosfomycin resistance among many pathogenic Gram-positive bacteria. In the present paper, we describe detailed kinetic studies of FosB from Staphylococcus aureus (SaFosB) that confirm that bacillithiol (BSH) is its preferred physiological thiol substrate. SaFosB is the first to be characterized among a new class of enzyme (bacillithiol-S-transferases), which, unlike glutathione transferases, are distributed among many low-G+C Gram-positive bacteria that use BSH instead of glutathione as their major low-molecular-mass thiol. The K(m) values for BSH and fosfomycin are 4.2 and 17.8 mM respectively. Substrate specificity assays revealed that the thiol and amino groups of BSH are essential for activity, whereas malate is important for SaFosB recognition and catalytic efficiency. Metal activity assays indicated that Mn(2+) and Mg(2+) are likely to be the relevant cofactors under physiological conditions. The serine analogue of BSH (BOH) is an effective competitive inhibitor of SaFosB with respect to BSH, but uncompetitive with respect to fosfomycin. Coupled with NMR characterization of the reaction product (BS-fosfomycin), this demonstrates that the SaFosB-catalysed reaction pathway involves a compulsory ordered binding mechanism with fosfomycin binding first followed by BSH which then attacks the more sterically hindered C-1 carbon of the fosfomycin epoxide. Disruption of BSH biosynthesis in S. aureus increases sensitivity to fosfomycin. Together, these results indicate that SaFosB is a divalent-metal-dependent bacillithiol-S-transferase that confers fosfomycin resistance on S. aureus.


Assuntos
Antibacterianos/química , Proteínas de Bactérias/química , Farmacorresistência Bacteriana , Fosfomicina/química , Staphylococcus aureus/enzimologia , Transferases/química , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cisteína/análogos & derivados , Cisteína/genética , Cisteína/metabolismo , Fosfomicina/farmacologia , Glucosamina/análogos & derivados , Glucosamina/genética , Glucosamina/metabolismo , Cinética , Magnésio/química , Magnésio/metabolismo , Manganês/química , Manganês/metabolismo , Staphylococcus aureus/genética , Transferases/genética , Transferases/metabolismo
15.
Biochem J ; 454(2): 239-47, 2013 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-23758290

RESUMO

BshB, a key enzyme in bacillithiol biosynthesis, hydrolyses the acetyl group from N-acetylglucosamine malate to generate glucosamine malate. In Bacillus anthracis, BA1557 has been identified as the N-acetylglucosamine malate deacetylase (BshB); however, a high content of bacillithiol (~70%) was still observed in the B. anthracis ∆BA1557 strain. Genomic analysis led to the proposal that another deacetylase could exhibit cross-functionality in bacillithiol biosynthesis. In the present study, BA1557, its paralogue BA3888 and orthologous Bacillus cereus enzymes BC1534 and BC3461 have been characterized for their deacetylase activity towards N-acetylglucosamine malate, thus providing biochemical evidence for this proposal. In addition, the involvement of deacetylase enzymes is also expected in bacillithiol-detoxifying pathways through formation of S-mercapturic adducts. The kinetic analysis of bacillithiol-S-bimane conjugate favours the involvement of BA3888 as the B. anthracis bacillithiol-S-conjugate amidase (Bca). The high degree of specificity of this group of enzymes for its physiological substrate, along with their similar pH-activity profile and Zn²âº-dependent catalytic acid-base reaction provides further evidence for their cross-functionalities.


Assuntos
Amidoidrolases/metabolismo , Bacillus anthracis/metabolismo , Bacillus cereus/metabolismo , Proteínas de Bactérias/metabolismo , Cisteína/análogos & derivados , Glucosamina/análogos & derivados , Acetilação , Acetilglucosamina/análogos & derivados , Acetilglucosamina/metabolismo , Amidoidrolases/química , Amidoidrolases/genética , Amidoidrolases/isolamento & purificação , Sequência de Aminoácidos , Substituição de Aminoácidos , Bacillus anthracis/enzimologia , Bacillus cereus/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Biocatálise , Domínio Catalítico , Sequência Conservada , Cisteína/metabolismo , Glucosamina/metabolismo , Concentração de Íons de Hidrogênio , Hidrólise , Malatos/metabolismo , Dados de Sequência Molecular , Proteínas Mutantes/química , Proteínas Mutantes/isolamento & purificação , Proteínas Mutantes/metabolismo , Filogenia , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Especificidade por Substrato , Zinco/metabolismo
16.
J Sci Med Sport ; 2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38777738

RESUMO

OBJECTIVES: To identify what coaches, support staff, and athletes believe should be considered when implementing menstrual cycle tracking in sport. DESIGN: Concept mapping (mixed-methods participatory approach). METHODS: Participants brainstormed statements in response to the focus prompt "What should be considered when tracking menstrual cycles in sport?" Participants then sorted the statements into groups according to meaning and rated each statement on its importance and feasibility to address. RESULTS: Twenty-six participants (12 athletes and 14 coaches/support staff) generated 53 ideas that the research team synthesised to 57 unique statements. Sixteen participants sorted the statements into an average of 8 (±4) groups. Six clusters were identified as the most appropriate representation of the sorting data following multidimensional scaling and hierarchical cluster analysis: 1) Ethics, 2) Tracking Tools and Methods, 3) Performance and Health, 4) Education, 5) Interpretation and Framing and 6) Communication. Participants rated each statement based on their importance (n = 12) and feasibility (n = 10), respectively. The most important (mean = 3.88 out of 5) and feasible (mean = 3.85 out of 5) cluster was Ethics, whilst the least important (mean = 3.17) and feasible (mean = 3.04) was Communication. CONCLUSIONS: Ethical considerations pertaining to privacy, consent, and scope of practice should be prioritised when tracking menstrual cycles in sport. A low burden method of tracking, which allows for some individualisation based on athletes' preferences or needs, and menstrual health education should also be undertaken to improve the uptake and impact of menstrual cycle tracking.

17.
Biochimie ; 220: 22-30, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38104714

RESUMO

Batrachochytrium dendrobatidis (Bd) is a lethal amphibian pathogen, partly due to its ability to evade the immune system of susceptible frog species. In many pathogenic fungi, the antioxidant glutathione is a virulence factor that helps neutralise oxidative stressors generated from host immune cells, as well as other environmental stressors such as heavy metals. The role of glutathione in stress tolerance in Bd has not been investigated. Here, we examine the changes in the glutathione pool after stress exposure and quantify the effect of glutathione depletion on cell growth and stress tolerance. Depletion of glutathione repressed growth and release of zoospores, suggesting that glutathione is essential for life cycle completion in Bd. Supplementation with <2 mM exogenous glutathione accelerated zoospore development, but concentrations >2 mM were strongly inhibitory to Bd cells. While hydrogen peroxide exposure lowered the total cellular glutathione levels by 42 %, glutathione depletion did not increase the sensitivity to hydrogen peroxide. Exposure to cadmium increased total cellular glutathione levels by 93 %. Glutathione-depleted cells were more sensitive to cadmium, and this effect was attenuated by glutathione supplementation, suggesting that glutathione plays an important role in cadmium tolerance. The effects of heat and salt were exacerbated by the addition of exogenous glutathione. The impact of glutathione levels on Bd stress sensitivity may help explain differences in host susceptibility to chytridiomycosis and may provide opportunities for synergistic therapeutics.


Assuntos
Batrachochytrium , Cádmio , Glutationa , Peróxido de Hidrogênio , Glutationa/metabolismo , Cádmio/toxicidade , Animais , Batrachochytrium/metabolismo , Peróxido de Hidrogênio/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Micoses/microbiologia , Micoses/veterinária , Micoses/metabolismo , Anfíbios/microbiologia
18.
Biochem Biophys Res Commun ; 436(2): 128-33, 2013 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-23618856

RESUMO

Bacillithiol (BSH), an α-anomeric glycoside of l-cysteinyl-d-glucosaminyl-l-malate, is a major low molecular weight thiol found in low GC Gram-positive bacteria, such as Staphylococcus aureus. Like other low molecular weight thiols, BSH is likely involved in protection against a number of stresses. We examined S. aureus transposon mutants disrupted in each of the three genes associated with BSH biosynthesis. These mutants are sensitive to alkylating stress, oxidative stress, and metal stress indicating that BSH and BSH-dependent enzymes are involved in protection of S. aureus. We further demonstrate that BshB, a deacetylase involved in the second step of BSH biosynthesis, also acts as a BSH conjugate amidase and identify S. aureus USA 300 LAC 2626 as a BSH-S-transferase, which is able to conjugate chlorodinitrobenzene, cerulenin, and rifamycin to BSH.


Assuntos
Cisteína/análogos & derivados , Glucosamina/análogos & derivados , Mutação , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Vias Biossintéticas/genética , Cromatografia Líquida de Alta Pressão , Cisteína/metabolismo , Glucosamina/metabolismo , Iodoacetamida/farmacologia , Metais/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/genética , Oxidantes/farmacologia , Aldeído Pirúvico/farmacologia , Staphylococcus aureus/enzimologia , Compostos de Sulfidrila/metabolismo , Fatores de Tempo
19.
Chembiochem ; 14(16): 2160-8, 2013 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-24115506

RESUMO

Bacillithiol (BSH) is the major low-molecular-weight (LMW) thiol in many low-G+C Gram-positive bacteria (Firmicutes). Evidence now emerging suggests that BSH functions as an important LMW thiol in redox regulation and xenobiotic detoxification, analogous to what is already known for glutathione and mycothiol in other microorganisms. The biophysical properties and cellular concentrations of such LMW thiols are important determinants of their biochemical efficiency both as biochemical nucleophiles and as redox buffers. Here, BSH has been characterised and compared with other LMW thiols in terms of its thiol pKa , redox potential and thiol-disulfide exchange reactivity. Both the thiol pKa and the standard thiol redox potential of BSH are shown to be significantly lower than those of glutathione whereas the reactivities of the two compounds in thiol-disulfide reactions are comparable. The cellular concentration of BSH in Bacillus subtilis varied over different growth phases and reached up to 5 mM, which is significantly greater than previously observed from single measurements taken during mid-exponential growth. These results demonstrate that the biophysical characteristics of BSH are distinctively different from those of GSH and that its cellular concentrations can reach levels much higher than previously reported.


Assuntos
Bacillus subtilis/química , Cisteína/análogos & derivados , Glucosamina/análogos & derivados , Aminas/química , Bacillus subtilis/metabolismo , Ácidos Carboxílicos/química , Cisteína/química , Glucosamina/química , Glutationa/química , Cinética , Oxirredução , Compostos de Sulfidrila/química
20.
J Funct Morphol Kinesiol ; 8(3)2023 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-37489302

RESUMO

Field lacrosse requires sudden directional changes and rapid acceleration/deceleration. The capacity to perform these skills is dependent on explosive muscle force production. Limited research exists on the potential of sprint interval training (SIT) to impact explosive muscle force production in field lacrosse players. The purpose of this study is to examine SIT, concurrent to field-lacrosse-specific training, on the rate of torque development (RTD), contractile impulse, and muscle function in female high school field lacrosse players (n = 12; 16 ± 1 yrs.). SIT was performed three times per week, concurrent to field-lacrosse-specific training, for 12 weeks. Right lower-limb muscle performance was assessed pre-, mid-, and post-SIT training via isometric and isokinetic concentric knee extensor contractions. Outcomes included RTD (Nm·s-1), contractile impulse (Nm·s), and peak torque (Nm). RTD for the first 50 ms of contraction improved by 42% by midseason and remained elevated at postseason (p = 0.004, effect size (ES) = -577.3 to 66.5). Contractile impulse demonstrated a training effect across 0-50 ms (42%, p = 0.004, ES = -1.4 to 0.4), 0-100 ms (33%, p = 0.018, ES = 3.1 to 0.9), and 0-200 ms (22%, p = 0.031, ES = -7.8 to 1.6). Isometric (0 rad·s-1) and concentric (3.1 rad·s-1) strength increased by 20% (p = 0.002, ES = -60.8 to -20.8) and 9% (p = 0.038, ES = -18.2 to 0.0) from SIT and field-lacrosse-specific training, respectively (p < 0.05). SIT, concurrent to field-lacrosse-specific training, enhanced lower-limb skeletal muscle performance, which may enable greater sport-specific gains.

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