ABSTRACT
With disease-modifying drugs on the horizon for degenerative ataxias, ecologically valid, finely granulated, digital health measures are highly warranted to augment clinical and patient-reported outcome measures. Gait and balance disturbances most often present as the first signs of degenerative cerebellar ataxia and are the most reported disabling features in disease progression. Thus, digital gait and balance measures constitute promising and relevant performance outcomes for clinical trials.This narrative review with embedded consensus will describe evidence for the sensitivity of digital gait and balance measures for evaluating ataxia severity and progression, propose a consensus protocol for establishing gait and balance metrics in natural history studies and clinical trials, and discuss relevant issues for their use as performance outcomes.
ABSTRACT
BACKGROUND: Mitochondrial disease is a heterogenous group of rare, complex neurometabolic disorders. Despite their individual rarity, collectively mitochondrial diseases represent the most common cause of inherited metabolic disorders in the UK; they affect 1 in every 4300 individuals, up to 15,000 adults (and a similar number of children) in the UK. Mitochondrial disease manifests multisystem and isolated organ involvement, commonly affecting those tissues with high energy demands, such as skeletal muscle. Myopathy manifesting as fatigue, muscle weakness and exercise intolerance is common and debilitating in patients with mitochondrial disease. Currently, there are no effective licensed treatments and consequently, there is an urgent clinical need to find an effective drug therapy. AIM: To investigate the efficacy of 12-week treatment with acipimox on the adenosine triphosphate (ATP) content of skeletal muscle in patients with mitochondrial disease and myopathy. METHODS: AIMM is a single-centre, double blind, placebo-controlled, adaptive designed trial, evaluating the efficacy of 12 weeks' administration of acipimox on skeletal muscle ATP content in patients with mitochondrial myopathy. Eligible patients will receive the trial investigational medicinal product (IMP), either acipimox or matched placebo. Participants will also be prescribed low dose aspirin as a non-investigational medical product (nIMP) in order to protect the blinding of the treatment assignment. Eighty to 120 participants will be recruited as required, with an interim analysis for sample size re-estimation and futility assessment being undertaken once the primary outcome for 50 participants has been obtained. Randomisation will be on a 1:1 basis, stratified by Fatigue Impact Scale (FIS) (dichotomised as < 40, ≥ 40). Participants will take part in the trial for up to 20 weeks, from screening visits through to follow-up at 16 weeks post randomisation. The primary outcome of change in ATP content in skeletal muscle and secondary outcomes relating to quality of life, perceived fatigue, disease burden, limb function, balance and walking, skeletal muscle analysis and symptom-limited cardiopulmonary fitness (optional) will be assessed between baseline and 12 weeks. DISCUSSION: The AIMM trial will investigate the effect of acipimox on modulating muscle ATP content and whether it can be repurposed as a new treatment for mitochondrial disease with myopathy. TRIAL REGISTRATION: EudraCT2018-002721-29 . Registered on 24 December 2018, ISRCTN 12895613. Registered on 03 January 2019, https://www.isrctn.com/search?q=aimm.
Subject(s)
Mitochondrial Myopathies , Muscular Diseases , Adult , Child , Humans , Adenosine Triphosphate , Aspirin/therapeutic use , Fatigue , Mitochondrial Myopathies/diagnosis , Mitochondrial Myopathies/drug therapy , Pyrazines , Quality of Life , Randomized Controlled Trials as TopicABSTRACT
Background and Aims: Gastrointestinal (GI) dysmotility is a common and debilitating clinical manifestation in patients with mitochondrial DNA (mtDNA)-related disease with no curative and few effective symptomatic therapies. A low-residue diet (LRD) has been shown to be effective at reducing bowel urgency, pain, and distension in functional GI-related conditions. We assessed tolerability and effects of an LRD on bowel habits in patients with mtDNA-related disease. Methods: This was a 12-week single-arm pilot study in patients with genetically determined primary mtDNA-related disease, meeting the ROME III constipation criteria. The co-primary outcomes were tolerability of an LRD (<10 g fiber per day) assessed by food diaries and changes in stool frequency and consistency. The secondary outcomes included GI symptoms, disease burden, laxatives, physical activity levels, colonic transit time using radiopaque markers, gut microbiome (patients and controls), and metabolomics. The gut microbiome of the mtDNA-related disease patients was compared against controls for observational purpose only. Results: Twenty-eight patients were enrolled, and 24 completed the LRD intervention. The LRD was well tolerated with a mean fold change of -34% in dietary fiber (5.3 ± 10.4 grams) per day (P = .03, confidence interval = 0.7-9.9) with no adverse events. The proportion of stool samples with normal stool consistency increased from 36% to 49% (P = .01); GI symptoms and laxative use were reduced. However, the LRD did not change stool frequency, stool output, and colonic transit time. The gut microbiome was significantly different between patients and controls but was not modulated by the dietary intervention. Conclusion: The LRD in patients with mtDNA-related mitochondrial disease and significant constipation is well tolerated and a promising treatment for alleviating GI symptoms. These positive findings should be confirmed in a randomized controlled trial.ClinicalTrials.gov Identifier: NCT03388528.
ABSTRACT
The quorum-sensing signaling systems in Vibrio bacteria converge to control levels of the master transcription factors LuxR/HapR, a family of highly conserved proteins that regulate gene expression for bacterial behaviors. A compound library screen identified 2-thiophenesulfonamide compounds that specifically inhibit Vibrio campbellii LuxR but do not affect cell growth. We synthesized a panel of 50 thiophenesulfonamide compounds to examine the structure-activity relationship effects on Vibrio quorum sensing. The most potent molecule identified, PTSP (3-phenyl-1-(thiophen-2-ylsulfonyl)-1H-pyrazole), inhibits quorum sensing in multiple strains of V. vulnificus, V. parahaemolyticus, and V. campbellii at nanomolar concentrations. However, thiophenesulfonamide inhibition efficacy varies significantly among Vibrio species: PTSP is most inhibitory against V. vulnificus SmcR, but V. cholerae HapR is completely resistant to all thiophenesulfonamides tested. Reverse genetics experiments show that PTSP efficacy is dictated by amino acid sequence in the putative ligand-binding pocket: F75Y and C170F SmcR substitutions are each sufficient to eliminate PTSP inhibition. Further, in silico modeling distinguished the most potent thiophenesulfonamides from less-effective derivatives. Our results revealed the previously unknown differences in LuxR/HapR proteins that control quorum sensing in Vibrio species and underscore the potential for developing thiophenesulfonamides as specific quorum sensing-directed treatments for Vibrio infections.
Subject(s)
Quorum Sensing/drug effects , Repressor Proteins/antagonists & inhibitors , Repressor Proteins/metabolism , Sulfonamides/metabolism , Sulfonamides/pharmacology , Trans-Activators/antagonists & inhibitors , Trans-Activators/metabolism , Vibrio/metabolism , Amino Acid Sequence , Amino Acid Substitution , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Ligands , Molecular Docking Simulation , Protein Binding , Protein Interaction Domains and Motifs , Repressor Proteins/chemistry , Species Specificity , Structure-Activity Relationship , Sulfonamides/chemistry , Trans-Activators/chemistry , Vibrio/chemistry , Vibrio/geneticsABSTRACT
BACKGROUND: The World Health Organization (WHO) recommends that people of all ages take regular and adequate physical activity. If unable to meet the recommendations due to health conditions, international guidance advises being as physically active as possible. Evidence from community interventions of physical activity indicate that people living with medical conditions are sometimes excluded from participation in studies. In this review, we considered the effects of activity-promoting interventions on physical activity and well-being in studies, as well as any adverse events experienced by participants living with inherited or acquired neuromuscular diseases (NMDs). OBJECTIVES: To assess the effects of interventions designed to promote physical activity in people with NMD compared with no intervention or alternative interventions. SEARCH METHODS: On 30 April 2020, we searched Cochrane Neuromuscular Specialised Register, CENTRAL, Embase, MEDLINE, and ClinicalTrials.Gov. WHO ICTRP was not accessible at the time. SELECTION CRITERIA: We considered randomised or quasi-randomised trials, including cross-over trials, of interventions designed to promote physical activity in people with NMD compared to no intervention or alternative interventions. We specifically included studies that reported physical activity as an outcome measure. Our main focus was studies in which promoting physical activity was a stated aim but we also included studies in which physical activity was assessed as a secondary or exploratory outcome. DATA COLLECTION AND ANALYSIS: We used standard Cochrane procedures. MAIN RESULTS: The review included 13 studies (795 randomised participants from 12 studies; number of participants unclear in one study) of different interventions to promote physical activity. Most studies randomised a minority of invited participants. No study involved children or adolescents and nine studies reported minimal entry criteria for walking. Participants had one of nine inherited or acquired NMDs. Types of intervention included structured physical activity support, exercise support (as a specific form of physical activity), and behaviour change support that included physical activity or exercise. Only one included study clearly reported that the aim of intervention was to increase physical activity. Other studies reported or planned to analyse the effects of intervention on physical activity as a secondary or exploratory outcome measure. Six studies did not report results for physical activity outcomes, or the data were not usable. We judged 10 of the 13 included studies at high or unclear risk of bias from incomplete physical activity outcome reporting. We did not perform a meta-analysis for any comparison because of differences in interventions and in usual care. We also found considerable variation in how studies reported physical activity as an outcome measure. The studies that reported physical activity measurement did not always clearly report intention-to-treat (ITT) analysis or whether final assessments occurred during or after intervention. Based on prespecified measures, we included three comparisons in our summary of findings. A physical activity programme (weight-bearing) compared to no physical activity programme One study involved adults with diabetic peripheral neuropathy (DPN) and reported weekly duration of walking during and at the end of a one-year intervention using a StepWatch ankle accelerometer. Based on the point estimate and low-certainty evidence, intervention may have led to an important increase in physical activity per week; however, the 95% confidence interval (CI) included the possibility of no difference or an effect in either direction at three months (mean difference (MD) 34 minutes per week, 95% CI -92.19 to 160.19; 69 participants), six months (MD 68 minutes per week, 95% CI -55.35 to 191.35; 74 participants), and 12 months (MD 49 minutes per week, 95% CI -75.73 to 173.73; 70 participants). Study-reported effect estimates for foot lesions and full-thickness ulcers also included the possibility of no difference, a higher, or lower risk with intervention. A sensor-based, interactive exercise programme compared to no sensor-based, interactive exercise programme One study involved adults with DPN and reported duration of walking over 48 hours at the end of four weeks' intervention using a t-shirt embedded PAMSys sensor. It was not possible to draw conclusions about the effectiveness of the intervention from the very low-certainty evidence (MD -0.64 hours per 48 hours, 95% CI -2.42 to 1.13; 25 participants). We were also unable to draw conclusions about impact on the Physical Component Score (PCS) for quality of life (MD 0.24 points, 95% CI -5.98 to 6.46; 35 participants; very low-certainty evidence), although intervention may have made little or no difference to the Mental Component Score (MCS) for quality of life (MD 5.10 points, 95% CI -0.58 to 10.78; 35 participants; low-certainty evidence). A functional exercise programme compared to a stretching exercise programme One study involved adults with spinal and bulbar muscular atrophy and reported a daily physical activity count at the end of 12 weeks' intervention using an Actical accelerometer. It was not possible to draw conclusions about the effectiveness of either intervention (requiring compliance) due to low-certainty evidence and unconfirmed measurement units (MD -8701, 95% CI -38,293.30 to 20,891.30; 43 participants). Functional exercise may have made little or no difference to quality of life compared to stretching (PCS: MD -1.10 points, 95% CI -5.22 to 3.02; MCS: MD -1.10 points, 95% CI -6.79 to 4.59; 49 participants; low-certainty evidence). Although studies reported adverse events incompletely, we found no evidence of supported activity increasing the risk of serious adverse events. AUTHORS' CONCLUSIONS: We found a lack of evidence relating to children, adolescents, and non-ambulant people of any age. Many people living with NMD did not meet randomised controlled trial eligibility criteria. There was variation in the components of supported activity intervention and usual care, such as physical therapy provision. We identified variation among studies in how physical activity was monitored, analysed, and reported. We remain uncertain of the effectiveness of promotional intervention for physical activity and its impact on quality of life and adverse events. More information is needed on the ITT population, as well as more complete reporting of outcomes. While there may be no single objective measure of physical activity, the study of qualitative and dichotomous change in self-reported overall physical activity might offer a pragmatic approach to capturing important change at an individual and population level.
Subject(s)
Exercise , Health Promotion/methods , Neuromuscular Diseases/rehabilitation , Bias , Humans , Muscle Stretching Exercises , Outcome Assessment, Health Care , Quality of Life , Randomized Controlled Trials as Topic , Resistance Training/statistics & numerical data , Time Factors , Walking/statistics & numerical dataABSTRACT
Quorum sensing gene expression in vibrios is regulated by the LuxR/HapR family of transcriptional factors, which includes Vibrio vulnificus SmcR. The consensus binding site of Vibrio LuxR/HapR/SmcR proteins is palindromic but highly degenerate with sequence variations at each promoter. To examine the mechanism by which SmcR recognizes diverse DNA sites, we generated SmcR separation-of-function mutants that either repress or activate transcription but not both. SmcR N55I is restricted in recognition of single base-pair variations in DNA binding site sequences and thus is defective at transcription activation but retains interaction with RNA polymerase (RNAP) alpha. SmcR S76A, L139R and N142D substitutions disrupt the interaction with RNAP alpha but retain functional DNA binding activity. X-ray crystallography and small angle X-ray scattering data show that the SmcR DNA binding domain exists in two conformations (wide and narrow), and the protein complex forms a mixture of dimers and tetramers in solution. The three RNAP interaction-deficient variants also have two DNA binding domain conformations, whereas SmcR N55I exhibits only the wide conformation. These data support a model in which two mechanisms drive SmcR transcriptional activation: interaction with RNAP and a multi-conformational DNA binding domain that permits recognition of variable DNA sites.
Subject(s)
Bacterial Proteins/chemistry , Bacterial Proteins/genetics , DNA-Binding Proteins/chemistry , Trans-Activators/chemistry , Trans-Activators/genetics , Transcription Factors/chemistry , Vibrio vulnificus/chemistry , Binding Sites , Crystallography, X-Ray , DNA-Binding Proteins/genetics , Dimerization , Gene Expression , Gene Expression Regulation, Bacterial/genetics , Models, Molecular , Mutation , Promoter Regions, Genetic , Protein Conformation , Quorum Sensing/genetics , Recombinant Proteins , Repressor Proteins/chemistry , Repressor Proteins/genetics , Scattering, Small Angle , Transcription Factors/genetics , Vibrio vulnificus/geneticsABSTRACT
Quorum sensing is a cell density-dependent form of cellular communication among bacteria. This signaling process has been heavily studied in vibrios due to their diverse and complex phenotypes and relevance to human and aquaculture disease. Mechanistic studies of Vibrio quorum sensing have required optimization of protein purification techniques to examine the role of key proteins, such as the LuxR/HapR family of transcription factors that control quorum-sensing gene expression. Protein purification is the cornerstone of biochemistry, and it is crucial to consistently produce batches of protein that are pure, active, and concentrated to perform various assays. The methods described here are optimized for purification of the Vibrio master quorum-sensing regulators, LuxR (Vibrio harveyi), HapR (Vibrio cholerae), and SmcR (Vibrio vulnificus). We anticipate that these methods can be applied to other proteins in this family of transcription factors.
Subject(s)
Bacterial Proteins/isolation & purification , Transcription Factors/isolation & purification , Vibrio/chemistry , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Quorum Sensing , Transcription Factors/chemistry , Transcription Factors/metabolismABSTRACT
Background: The benefit and safety of exercise training for patients with neuromuscular disorders (NMDs) has long been a contentious topic. This is, in part, due to recognised challenges associated with rare diseases including small and heterogenous patient populations. We performed a systematic review and meta-analyses to evaluate the effectiveness and safety of interventional exercise and establish minimal clinically important differences (MCID) in outcomes to facilitate clinical interpretation. Methods: We searched six databases from inception to Mar 2018. Aerobic, strength, and combined (aerobic and strength) intervention were eligible. Meta-analyses compared outcomes at baseline with those after at least six weeks (before-after exercise within individuals). A further meta-analysis compared outcomes before-after exercise between groups (exercise training versus usual care). Disease heterogeneity was explored using a random effect model. This study was registered (PROSPERO, CRD42018102183). An interactive database was developed to facilitate full interrogations of data. Results: We identified 130 articles describing 1,805 participants with 35 different forms of NMD. Of these studies, 76 were suitable for meta-analyses. Within group and between group meta-analyses detected an increase in peak aerobic capacity (p=0·04), and peak power (p=0·01). Six-minute walk test (p=0·04), sit-to-stand (STS) (repetitions) (p=0·03), STS (seconds) (p=0·04), rise from supine (p=0·008), SF-36 (p=0·0003), fatigue severity (p=<0·0001), citrate synthase (p=0·0002), central nuclei (p=0·04), type 1 (p=0·002) and type II muscle fibre area (p=0·003), were only able to detect change within group meta-analyses. Substantial I 2 statistic heterogeneity was revealed for STS (seconds) ( I²=58·5%; p=0·04) and citrate synthase ( I²=70·90%; p=0·002), otherwise heterogeneity for all outcomes was low. No study-related serious adverse events were reported nor significant increases in creatine kinase. Conclusions: Exercise training in patients with NMDs appears to cause no harm across a range of outcomes. With the emergence of new therapeutic strategies, defining MCID is vital in informing future clinical trial design.
ABSTRACT
The marine facultative pathogen Vibrio cholerae forms complex multicellular communities on the chitinous shells of crustacean zooplankton in its aquatic reservoir. V. cholerae-chitin interactions are critical for the growth, evolution, and waterborne transmission of cholera. This is due, in part, to chitin-induced changes in gene expression in this pathogen. Here, we sought to identify factors that influence chitin-induced expression of one locus, the chitobiose utilization operon (chb), which is required for the uptake and catabolism of the chitin disaccharide. Through a series of genetic screens, we identified that the master regulator of quorum sensing, HapR, is a direct repressor of the chb operon. We also found that the levels of HapR in V. cholerae are regulated by the ClpAP protease. Furthermore, we show that the canonical quorum sensing cascade in V. cholerae regulates chb expression in an HapR-dependent manner. Through this analysis, we found that signaling via the species-specific autoinducer CAI-1, but not the interspecies autoinducer AI-2, influences chb expression. This phenomenon of species-specific regulation may enhance the fitness of this pathogen in its environmental niche.IMPORTANCE In nature, bacteria live in multicellular and multispecies communities. Microbial species can sense the density and composition of their community through chemical cues using a process called quorum sensing (QS). The marine pathogen Vibrio cholerae is found in communities on the chitinous shells of crustaceans in its aquatic reservoir. V. cholerae interactions with chitin are critical for the survival, evolution, and waterborne transmission of this pathogen. Here, we show that V. cholerae uses QS to regulate the expression of one locus required for V. cholerae-chitin interactions.
Subject(s)
Bacterial Proteins/genetics , Disaccharides/metabolism , Operon , Quorum Sensing , Vibrio cholerae/genetics , Bacterial Proteins/metabolism , Species Specificity , Vibrio cholerae/metabolismABSTRACT
Mitochondrial disorders affect 1/5,000 and have no cure. Inducing mitochondrial biogenesis with bezafibrate improves mitochondrial function in animal models, but there are no comparable human studies. We performed an open-label observational experimental medicine study of six patients with mitochondrial myopathy caused by the m.3243A>G MTTL1 mutation. Our primary aim was to determine the effects of bezafibrate on mitochondrial metabolism, whilst providing preliminary evidence of safety and efficacy using biomarkers. The participants received 600-1,200 mg bezafibrate daily for 12 weeks. There were no clinically significant adverse events, and liver function was not affected. We detected a reduction in the number of complex IV-immunodeficient muscle fibres and improved cardiac function. However, this was accompanied by an increase in serum biomarkers of mitochondrial disease, including fibroblast growth factor 21 (FGF-21), growth and differentiation factor 15 (GDF-15), plus dysregulation of fatty acid and amino acid metabolism. Thus, although potentially beneficial in short term, inducing mitochondrial biogenesis with bezafibrate altered the metabolomic signature of mitochondrial disease, raising concerns about long-term sequelae.
Subject(s)
Bezafibrate/pharmacology , Mitochondria/metabolism , Mitochondrial Myopathies/drug therapy , Humans , Mitochondrial Myopathies/metabolism , Organelle BiogenesisABSTRACT
BACKGROUND: Academic surgeons are encouraged to promote their work on social media. We hypothesized that thoracic surgeons who are active on Twitter have a higher research citation index (Hirsch index [h-index]) than their counterparts who are not. METHODS: Thoracic surgeons on CTSNet.org in Canada and the United States were queried for profiles with an h-index on Google Scholar and/or Research Gate in July 2018. Surgeons were categorized by whether they possessed a Twitter account (T+) or not (T-), and h-index values were compared. Within the T+ cohort a multivariate regression model was used to identify independent predictors of increased h-index among variables related to Twitter activity. RESULTS: Of 3741 surgeons queried, 19.3% (722) had a known h-index. The mean h-index for the entire cohort was 14.54 (SD, 15.73). The median h-index was 10 (range, 0-121), and the 75th percentile h-index was 20. T+ surgeons had a median h-index of 10 (range, 0-66), and T- surgeons had a median h-index of 10 (range, 0-72; P = .25). The 75th percentile h-index for T+ surgeons was 23 compared with 20 for T- surgeons (P = .24). For T+ surgeons the regression model identified the number of followers (P = .029), the number of people followed (P = .048), and the frequency of tweeting (P = .046) as independent predictors of a higher h-index. CONCLUSIONS: The median h-index for an academic thoracic surgeon in Canada and the United States is 10. Surgeons who engage in Twitter activity are more likely to have their research cited by others.
Subject(s)
Biomedical Research/methods , Periodicals as Topic/statistics & numerical data , Social Media/statistics & numerical data , Surgeons/statistics & numerical data , Thoracic Surgery , Canada , Humans , United StatesABSTRACT
OBJECTIVES: Defining clinically relevant outcome measures for myotonic dystrophy type 1 (DM1) that can be valid and feasible for different phenotypes has proven problematic. The Outcome Measures for Myotonic Dystrophy (OMMYD) group proposed a battery of functional outcomes: 6-minute walk test, 30 seconds sit and stand test, timed 10 m walk test, timed 10 m walk/run test, and nine-hole peg test. This, however, required a large-scale investigation, METHODS: A cohort of 213 patients enrolled in the natural history study, PhenoDM1, was analyzed in cross-sectional analysis and subsequently 98 patients were followed for longitudinal analysis. We aimed to assess: (1) feasibility and best practice; (2) intra-session reliability; (3) validity; and (4) behavior over time, of these tests. RESULTS: OMMYD outcomes proved feasible as 96% of the participants completed at least one trial for all tests and more than half (n = 113) performed all three trials of each test. Body mass index and disease severity associate with functional capacity. There was a significant difference between the first and second trials of each test. There was a moderate to strong correlation between these functional outcomes and muscle strength, disease severity and patient-reported outcomes. All outcomes after 1 year detected a change in functional capacity except the nine-hole peg test. CONCLUSIONS: These tests can be used as a battery of outcomes or independently based on the shown overlapping psychometric features and strong cross-correlations. Due to the large and heterogeneous sample of this study, these results can serve as reference values for future studies.
Subject(s)
Muscle Strength/physiology , Myotonic Dystrophy/physiopathology , Walk Test/methods , Adult , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Outcome Assessment, Health Care , Severity of Illness IndexABSTRACT
The objective of this study was to assess the impact of in-feed flavophospholipol on Salmonella shedding and antibody response in nursery pigs. Weaned pigs were fed either a diet containing 4 ppm flavophospholipol (n = 16) or a non-medicated feed (n = 16) for 36 d. All pigs were orally challenged with a 2-mL dose of 108 colony-forming units (CFUs)/mL of Salmonella Typhimurium on Days 7 and 8 of the trial. On Day 36, all pigs were euthanized and samples were collected from the liver, spleen, and ileocecal lymph nodes. Fecal and tissue samples were quantitatively cultured for Salmonella and serum samples were tested for the presence of the Salmonella antibody by enzyme-linked immunosorbent assay (ELISA). There was no difference between the 2 groups in antibody response and the presence of Salmonella in feces and tissue (P > 0.05). Medicating nursery diets with flavophospholipol at 4 ppm did not appear to reduce Salmonella infection in nursery pigs.
L'objectif de la présente étude était d'évaluer l'impact de l'ajout de flavophospholipol dans l'aliment sur l'excrétion de Salmonella et la réponse en anticorps chez des porcs en pouponnière. Des porcs sevrés ont été nourris avec soit une diète contenant 4 ppm de flavophospholipol (n = 16) ou une diète non-médicamentée (n = 16) pendant 36 j. Tous les porcs ont reçu oralement une dose de 2 mL de 108 unités formatrices de colonies (UFC)/mL de Salmonella Typhimurium aux Jours 7 et 8 de l'essai. Au Jour 36, tous les porcs ont été euthanasiés et on préleva des échantillons de foie, rate, et noeuds lymphatiques iléo-caecaux. Des échantillons de fèces et de tissus ont été cultivés pour quantifier le nombre de Salmonella et des échantillons de sérum furent testés pour la présence d'anticorps contre Salmonella par épreuve immunoenzymatique (ELISA). Il n'y avait pas de différence entre les deux groupes quant à la réponse en anticorps et la présence de Salmonella dans les fèces et les tissus (P > 0,05). L'ajout de 4 ppm de flavophospholipol à la diète en pouponnière ne semble pas réduire l'infection par Salmonella chez les porcs en pouponnière.(Traduit par Docteur Serge Messier).
Subject(s)
Animal Feed/analysis , Bacterial Shedding/drug effects , Bambermycins/pharmacology , Salmonella Infections, Animal/prevention & control , Salmonella typhimurium , Swine Diseases/microbiology , Animals , Diet/veterinary , Swine , Swine Diseases/prevention & controlABSTRACT
Salmonella is an important cause of foodborne illnesses in humans. Food-producing animals, including swine, are a major source of Salmonella in food products. This study investigated on farm Salmonella fecal shedding in pigs at different production stages - from weaning to marketing - and its association with the presence of Salmonella in tissues at slaughter. Fourteen groups from 8 commercial farrowing sources (N = 809 pigs) were monitored 5 times from birth to slaughter. Fecal and tissue samples were collected from pigs and cultured for Salmonella. A survey was conducted to collect farm management information. A multi-level mixed-effects logistic regression modelling method was used to analyze Salmonella shedding over time and the association between Salmonella shedding and the presence of Salmonella in tissue samples. Salmonella was recovered from 13% (421/3339) of fecal samples collected from 809 pigs over the course of the study. Overall, 35% (284) of pigs shed Salmonella at least once, while 12% (99) shed more than once. Salmonella shedding increased as pigs aged (P = 0.01) and increased in the summer months (P < 0.01). Salmonella was isolated from tissue samples collected from 23% (134/580) of pigs; however, the presence of Salmonella at slaughter was not associated with on farm shedding. The seasonal trend in Salmonella shedding and its association with age may be used to identify high-risk groups and implement more effective control measures accordingly. The identification of repeat shedders warrants interventions that target this source of infection on swine farms.
Salmonella est une cause importante de maladies d'origine alimentaire chez les humains. Les animaux de rente, incluant le porc, sont une source majeure de Salmonella dans les produits alimentaires. Au cours de la présente étude nous avons examiné l'excrétion fécale de Salmonella à la ferme à différents stades de la production du sevrage jusqu'à la mise en marché et sont association avec la présence de Salmonella dans les tissus au moment de l'abattage. Quatorze groupes provenant de huit sources commerciales de mise-bas (N = 809 porcs) ont été surveillés cinq fois entre la naissance et l'abattage. Des échantillons de fèces et de tissus ont été prélevés des porcs et cultivés pour Salmonella. Un sondage a été mené pour amasser des informations sur la gestion de la ferme. Une méthode de modélisation de régression logistique à effets mixtes de niveaux multiples a été utilisée pour analyser l'excrétion de Salmonella dans le temps et l'association entre l'excrétion de Salmonella et la présence de Salmonella dans les échantillons de tissus. Salmonella a été isolé de 13 % (421/3339) des échantillons de fèces prélevés des 809 porcs durant la durée de cette étude. Au total, 35 % (284) des porcs ont excrété Salmonella au moins une fois, alors que 12 % (99) ont excrété plus d'une fois. L'excrétion de Salmonella augmentait à mesure que les porcs vieillissaient (P = 0,01) et augmentait durant les mois d'été (P < 0,01). Salmonella a été isolé d'échantillons de tissu prélevés de 23 % (134/580) des porcs; toutefois, la présence de Salmonella au moment de l'abattage n'était pas associée avec l'excrétion à la ferme. La tendance saisonnière dans l'excrétion de Salmonella et son association avec l'âge pourraient être utilisées afin d'identifier les groupes à risque élevé et mettre en place selon le cas des méthodes de maitrise plus efficaces. L'identification d'excréteurs à répétition justifie des interventions qui ciblent cette source d'infection sur les fermes porcines.(Traduit par Docteur Serge Messier).
Subject(s)
Bacterial Shedding , Feces/microbiology , Lymph Nodes/microbiology , Salmonella Infections, Animal/microbiology , Salmonella/isolation & purification , Swine Diseases/microbiology , Animals , Ontario/epidemiology , Risk Factors , Salmonella Infections, Animal/epidemiology , Swine , Swine Diseases/epidemiologyABSTRACT
OBJECTIVE: Single, large-scale deletions in mitochondrial DNA (mtDNA) are a common cause of mitochondrial disease. This study aimed to investigate the relationship between the genetic defect and molecular phenotype to improve understanding of pathogenic mechanisms associated with single, large-scale mtDNA deletions in skeletal muscle. METHODS: We investigated 23 muscle biopsies taken from adult patients (6 males/17 females with a mean age of 43 years) with characterized single, large-scale mtDNA deletions. Mitochondrial respiratory chain deficiency in skeletal muscle biopsies was quantified by immunoreactivity levels for complex I and complex IV proteins. Single muscle fibers with varying degrees of deficiency were selected from 6 patient biopsies for determination of mtDNA deletion level and copy number by quantitative polymerase chain reaction. RESULTS: We have defined 3 "classes" of single, large-scale deletion with distinct patterns of mitochondrial deficiency, determined by the size and location of the deletion. Single fiber analyses showed that fibers with greater respiratory chain deficiency harbored higher levels of mtDNA deletion with an increase in total mtDNA copy number. For the first time, we have demonstrated that threshold levels for complex I and complex IV deficiency differ based on deletion class. INTERPRETATION: Combining genetic and immunofluorescent assays, we conclude that thresholds for complex I and complex IV deficiency are modulated by the deletion of complex-specific protein-encoding genes. Furthermore, removal of mt-tRNA genes impacts specific complexes only at high deletion levels, when complex-specific protein-encoding genes remain. These novel findings provide valuable insight into the pathogenic mechanisms associated with these mutations. Ann Neurol 2018;83:115-130.
Subject(s)
DNA, Mitochondrial/genetics , Mitochondrial Diseases/genetics , Sequence Deletion/genetics , Adult , Aged , Biopsy , Cohort Studies , Electron Transport Complex I/genetics , Electron Transport Complex IV/genetics , Female , Gene Deletion , Gene Dosage , Humans , Male , Middle Aged , Mitochondrial Diseases/pathology , Muscle Fibers, Skeletal/pathology , Muscle, Skeletal/pathology , Oxidative Phosphorylation , Young AdultABSTRACT
OBJECTIVE: Exercise intolerance is a clinical hallmark of chronic conditions. The present study determined pathophysiological mechanisms of exercise intolerance in cardiovascular, neuromuscular, and metabolic disorders. METHODS: In a prospective cross-sectional observational study 152 patients (heart failure reduced ejection fraction, n=32; stroke, n=34; mitochondrial disease, n=28; type two diabetes, n=28; and healthy controls, n=30) performed cardiopulmonary exercise testing with metabolic and haemodynamic measurements. Peak exercise O2 consumption and cardiac power output were measures of exercise tolerance and cardiac performance. RESULTS: Exercise tolerance was significantly diminished in patients compared with controls (ie, by 45% stroke, 39% mitochondria disease, and 33% diabetes and heart failure, p<0.05). Cardiac performance was only significantly reduced in heart failure (due to reduced heart rate, stroke volume, and blood pressure) and mitochondrial patients (due reduced stroke volume) compared with controls (ie, by 53% and 26%, p<0.05). Ability of skeletal muscles to extract oxygen (ie, arterial-venous O2 difference) was diminished in mitochondrial, stroke, and diabetes patients (by 24%, 22%, and 18%, p<0.05), but increased by 21% in heart failure (p<0.05) compared with controls. Cardiac output explained 65% and 51% of the variance in peak O2 consumption (p<0.01) in heart failure and mitochondrial patients, whereas arterial-venous O2 difference explained 69% (p<0.01) of variance in peak O2 consumption in diabetes, and 65% and 48% in stroke and mitochondrial patients (p<0.01). CONCLUSIONS: Different mechanisms explain exercise intolerance in patients with heart failure, mitochondrial dysfunction, stroke and diabetes. Their better understanding may improve management of patients, their stress tolerance and quality of life.
ABSTRACT
BACKGROUND: Free-living or habitual physical activity (HPA) refers to someone's performance in his or her free-living environment. Neuromuscular disorders (NMD) manifest through HPA, and the observation of HPA can be used to identify clinical risks and to quantify outcomes in research. This review summarizes and analyses previous studies reporting the assessment of HPA in NMD, and may serve as the basis for evidence-based decision-making when considering assessing HPA in this population. METHODS: A systematic review was performed to identify all studies related to HPA in NMD, followed by a critical appraisal of the assessment methodology and a final review of the identified HPA tools. RESULTS: A total of 22 studies were selected, reporting on eight different direct tools (or activity monitors) and ten structured patient-reported outcomes. Overall, HPA patterns in NMD differ from healthy control populations. There was a noticeable lack of validation studies for these tools and outcome measures in NMD. Very little information regarding feasibility and barriers for the application of these tools in this population have been published. CONCLUSIONS: The variety and heterogeneity of tools and methods in the published literature makes the comparison across different studies difficult, and methodological guidelines are warranted. We propose a checklist of considerations for the assessment and reporting of HPA in NMD.
Subject(s)
Exercise , Fitness Trackers , Habits , Neuromuscular Diseases/physiopathology , Patient Reported Outcome Measures , Case-Control Studies , Humans , Surveys and QuestionnairesABSTRACT
Perceived fatigue is a prominent symptom in patients with mitochondrial disease but to date its prevalence, impact and aetiology are poorly understood. Our aim was to determine the prevalence and assess for comorbidities associated with clinically relevant fatigue in patients with mitochondrial disease. A cross-sectional postal survey of patients with mitochondrial disease was undertaken using a validated self-completion, patient-reported outcome measures (response rate: 60%; n = 132). The prevalence and perceived functional impact of experienced fatigue were assessed using the Fatigue Impact Scale. Other putative biological mechanisms were evaluated using the Hospital Anxiety Depression scale and Epworth sleepiness scale. Data were compared with those for healthy control subjects and patients with Myalgic Encephalopathy/Chronic Fatigue Syndrome matched for age and gender. Sixty-two per cent of patients with mitochondrial disease reported excessive symptomatic fatigue (Fatigue Impact Scale ≥ 40); whilst 32% reported severe, functionally limiting fatigue symptoms (Fatigue Impact Scale ≥ 80) comparable to perceived fatigue in patients with Myalgic Encephalopathy/Chronic Fatigue Syndrome. Fatigue is common and often severe in patients with mitochondrial disease irrespective of age, gender or genotype. Future evaluation of causal factors in mitochondrial disease-associated fatigue is warranted with the potential to guide future treatment modalities.
Subject(s)
Fatigue/epidemiology , Mitochondrial Diseases/epidemiology , Adolescent , Adult , Aged , Aged, 80 and over , Disorders of Excessive Somnolence/epidemiology , Disorders of Excessive Somnolence/genetics , Disorders of Excessive Somnolence/physiopathology , Fatigue/genetics , Fatigue/physiopathology , Female , Genotype , Humans , Male , Middle Aged , Mitochondrial Diseases/genetics , Mitochondrial Diseases/physiopathology , Mitochondrial Diseases/psychology , Muscle Weakness/epidemiology , Muscle Weakness/genetics , Muscle Weakness/physiopathology , Perception , Prevalence , Severity of Illness Index , Surveys and Questionnaires , Young AdultABSTRACT
Complex I (CI) is the largest of the five multi-subunit complexes constituting the human oxidative phosphorylation (OXPHOS) system. Seven of its catalytic core subunits are encoded by mitochondrial DNA (ND (NADH dehydrogenase)1-6, ND4L (NADH dehydrogenase subunit 4L)), with mutations in all seven having been reported in association with isolated CI deficiency. We investigated two unrelated adult patients presenting with marked exercise intolerance, persistent lactic acidaemia and severe muscle-restricted isolated CI deficiency associated with sub-sarcolemmal mitochondrial accumulation. Screening of the mitochondrial genome detected novel mutations in the MTND1 (NADH dehydrogenase subunit 1) gene, encoding subunit of CI [Patient 1, m.3365T>C predicting p.(Leu20Pro); Patient 2, m.4175G>A predicting p.(Trp290*)] at high levels of mitochondrial DNA heteroplasmy in skeletal muscle. We evaluated the effect of these novel MTND1 mutations on complex assembly showing that CI assembly, although markedly reduced, was viable in the absence of detectable ND1 signal. Real-time PCR and Western blotting showed overexpression of different CI assembly factor transcripts and proteins in patient tissue. Together, our data indicate that the mechanism underlying the expression of the biochemical defect may involve a compensatory response to the novel MTND1 gene mutations, promoting assembly factor up-regulation and stabilization of respiratory chain super-complexes, resulting in partial rescue of the clinical phenotype.