Our current understanding of the toxicity of altered mito-ribosomal fidelity during mitochondrial protein synthesis: What can it tell us about human disease?

Altered mito-ribosomal fidelity is an important and insufficiently understood causative agent of mitochondrial dysfunction. Its pathogenic effects are particularly well-known in the case of mitochondrially induced deafness, due to the existence of the, so called, ototoxic variants at positions 847C (m.1494C) and 908A (m.1555A) of 12S mitochondrial (mt-) rRNA. It was shown long ago that the deleterious effects of these variants could remain dormant until an external stimulus triggered their pathogenicity. Yet, the link from the fidelity defect at the mito-ribosomal level to its phenotypic manifestation remained obscure. Recent work with fidelity-impaired mito-ribosomes, carrying error-prone and hyper-accurate mutations in mito-ribosomal proteins, have started to reveal the complexities of the phenotypic manifestation of mito-ribosomal fidelity defects, leading to a new understanding of mtDNA disease. While much needs to be done to arrive to a clear picture of how defects at the level of mito-ribosomal translation eventually result in the complex patterns of disease observed in patients, the current evidence indicates that altered mito-ribosome function, even at very low levels, may become highly pathogenic. The aims of this review are three-fold. First, we compare the molecular details associated with mito-ribosomal fidelity to those of general ribosomal fidelity. Second, we gather information on the cellular and organismal phenotypes associated with defective translational fidelity in order to provide the necessary grounds for an understanding of the phenotypic manifestation of defective mito-ribosomal fidelity. Finally, the results of recent experiments directly tackling mito-ribosomal fidelity are reviewed and future paths of investigation are discussed.

Structural analysis of mitochondrial rRNA gene variants identified in patients with deafness.

The last few years have witnessed dramatic advances in our understanding of the structure and function of the mammalian mito-ribosome. At the same time, the first attempts to elucidate the effects of mito-ribosomal fidelity (decoding accuracy) in disease have been made. Hence, the time is right to push an important frontier in our understanding of mitochondrial genetics, that is, the elucidation of the phenotypic effects of mtDNA variants affecting the functioning of the mito-ribosome. Here, we have assessed the structural and functional role of 93 mitochondrial (mt-) rRNA variants thought to be associated with deafness, including those located at non-conserved positions. Our analysis has used the structural description of the human mito-ribosome of the highest quality currently available, together with a new understanding of the phenotypic manifestation of mito-ribosomal-associated variants. Basically, any base change capable of inducing a fidelity phenotype may be considered non-silent. Under this light, out of 92 previously reported mt-rRNA variants thought to be associated with deafness, we found that 49 were potentially non-silent. We also dismissed a large number of reportedly pathogenic mtDNA variants, 41, as polymorphisms. These results drastically update our view on the implication of the primary sequence of mt-rRNA in the etiology of deafness and mitochondrial disease in general. Our data sheds much-needed light on the question of how mt-rRNA variants located at non-conserved positions may lead to mitochondrial disease and, most notably, provide evidence of the effect of haplotype context in the manifestation of some mt-rRNA variants.

The Identification of Multidrug-Resistant Microorganisms including Bergeyella zoohelcum Acquired from the Skin/Prosthetic Interface of Amputees and Their Susceptibility to Medihoney™ and Garlic Extract (Allicin).

Users of prosthetic devices face the accumulation of potentially drug-resistant pathogenic bacteria on the skin/prosthesis interface. In this study, we took surface swabs of the skin/prosthesis interface of eleven disabled athletes to identify microorganisms present. In addition to determining their antimicrobial resistance profile, we assessed their sensitivity to Manuka honey and Garlic extract (allicin). Eleven volunteers were directed to swab the skin at the skin/prosthesis interface. After initial isolation of microorganisms, we employed the following general microbiological methods: Gram stain, Catalase test, Oxidase test, lactose fermenting capability, haemolytic capability, Staphaurex, mannitol fermenting capability, Streptex; API Staph, 20E, Candida, and BBL crystal identification system tests. Once identified, isolates were analysed for their sensitivity to penicillin, erythromycin, ampicillin, vancomycin, ceftazidime, ciprofloxacin, gentamicin, and colistin-sulphate. Isolates were also analysed for their sensitivity to allicin (Garlic Extract (GE)) and Manuka honey (Medihoney™) (MH). Eleven isolates were identified: Bacillus cereus, Staphylococcus haemolyticus, Staphylococcus aureus, Micrococcus luteus, Pseudomonas oryzihabitans, Micrococcus spp., Bacillus subtilis, Group D Streptococcus, Pantoea spp., Enterobacter cloacae, and Bergeyella zoohelcum. All isolates were resistant to 1 unit of penicillin and 10 µg of ampicillin. Bergeyella zoohelcum was observed to have the widest range of resistance with observed resistance against five of the eight antimicrobials employed in this study. This study highlights the prevalence of uncommon drug-resistant microorganisms on the skin within a vulnerable population, highlighting the potential for MH or GE intervention.

The Science of Handcycling: A Narrative Review.

The aim of this narrative review is to provide insight as to the history, biomechanics, and physiological characteristics of competitive handcycling. Furthermore, based upon the limited evidence available, this paper aims to provide practical training suggestions by which to develop competitive handcycling performance. Handbike configuration, individual physiological characteristics, and training history all play a significant role in determining competitive handcycling performance. Optimal handcycling technique is highly dependent upon handbike configuration. As such, seat positioning, crank height, crank fore-aft position, crank length, and handgrip position must all be individually configured. In regard to physiological determinants, power output at a fixed blood lactate concentration of 4 mmol·L-1, relative oxygen consumption, peak aerobic power output, relative upper body strength, and maximal anaerobic power output have all been demonstrated to impact upon handcycling performance capabilities. Therefore, it is suggested that that an emphasis be placed upon the development and frequent monitoring of these parameters. Finally, linked to handcycling training, it is suggested that handcyclists should consider adopting a concurrent strength and endurance training approach, based upon a block periodization model that employs a mixture of endurance, threshold, interval, and strength training sessions. Despite our findings, it is clear that several gaps in our scientific knowledge of handcycling remain and that further research is necessary in order to improve our understanding of factors that determine optimal performance of competitive handcyclists. Finally, further longitudinal research is required across all classifications to study the effects of different training programs upon handcycling performance.

Perceived barriers to, and benefits of physical activity among British military veterans that are wounded, injured, and/or sick: a Behaviour Change Wheel perspective.

PURPOSE: Many British military veterans that are wounded, injured, and/or sick (WIS) face significant physical, psychological and social challenges following discharge from the military. There is increasing evidence to support the application and benefit of physical activity for veterans that are WIS. Understanding engagement in physical activity is therefore imperative so that physical activity interventions and initiatives can be designed effectively, and their benefits optimised. Therefore, the aim of this study is to identify the perceived barriers to, and benefits of physical activity among veterans that are WIS. MATERIALS AND METHODS: Nine semi-structured interviews were conducted to explore perceived barriers to, and benefits of physical activity amongst veterans that are WIS. A thematic analysis was conducted, and themes mapped to the Behaviour Change Wheel's capability, opportunity, motivation-behaviour (COM-B) model. RESULTS: Perceptions related to physical capability, psychological capability, physical opportunity and reflective motivation were predominant barriers to engagement in physical activity. As well providing opportunities to socialise, the perceived benefits of engagement in physical activity seemed to mirror the perceived barriers, suggesting a reciprocal relationship. CONCLUSIONS: Using the Behaviour Change Wheel, incentivisation, education, persuasion, enablement and environmental restructuring were identified as potentially beneficial intervention functions when seeking to increase levels of physical activity among veterans that are WIS living within the United Kingdom.IMPLICATIONS FOR REHABILITATIONUK-based veterans that are wounded, injured, and/or sick (WIS) often face significant challenges with respect to physical, mental, and social well-being.Perceived barriers, in the form of limited opportunities and a self-perceived lack of capability impact on levels of motivation and are viewed as obstacles to becoming physically active by veterans that are WIS.Providing education and complementary incentives may help reduce the extent of perceived barriers in veterans that are WIS.Approaches that allow veterans that are WIS to socialise while developing self-efficacy will likely increase physical activity engagement.

Heterologous Inferential Analysis (HIA) and Other Emerging Concepts: In Understanding Mitochondrial Variation In Pathogenesis: There is no More Low-Hanging Fruit.

Here we summarize our latest efforts to elucidate the role of mtDNA variants affecting the mitochondrial translation machinery, namely variants mapping to the mt-rRNA and mt-tRNA genes. Evidence is accumulating to suggest that the cellular response to interference with mitochondrial translation is different from that occurring as a result of mutations in genes encoding OXPHOS proteins. As a result, it appears safe to state that a complete view of mitochondrial disease will not be obtained until we understand the effect of mt-rRNA and mt-tRNA variants on mitochondrial protein synthesis. Despite the identification of a large number of potentially pathogenic variants in the mitochondrially encoded rRNA (mt-rRNA) genes, we lack direct methods to firmly establish their pathogenicity. In the absence of such methods, we have devised an indirect approach named heterologous inferential analysis (HIA ) that can be used to make predictions concerning the disruptive potential of a large subset of mt-rRNA variants. We have used HIA to explore the mutational landscape of 12S and 16S mt-rRNA genes. Our HIA studies include a thorough classification of all rare variants reported in the literature as well as others obtained from studies performed in collaboration with physicians. HIA has also been used with non-mammalian mt-rRNA genes to elucidate how mitotypes influence the interaction of the individual and the environment. Regarding mt-tRNA variations, rapidly growing evidence shows that the spectrum of mutations causing mitochondrial disease might differ between the different mitochondrial haplogroups seen in human populations.

The Relationship Between Absolute and Relative Upper-Body Strength and Handcycling Performance Capabilities.

PURPOSE: To explore the relationship between absolute and relative upper-body strength and selected measures of handcycling performance. METHODS: A total of 13 trained H3/H4-classified male handcyclists (mean [SD] age 37 [11] y; body mass 76.6 [10.1] kg; peak oxygen consumption 2.8 [0.6] L·min-1; relative peak oxygen consumption 36.5 [10] mL·kg·min-1) performed a prone bench-pull and bench-press 1-repetition-maximum strength assessment, a 15-km individual time trial, a graded exercise test, and a 15-second all-out sprint test. Relationships between all variables were assessed using Pearson correlation coefficient. RESULTS: Absolute strength measures displayed a large correlation with gross mechanical efficiency and maximum anaerobic power output (P = .05). However, only a small to moderate relationship was identified with all other measures. In contrast, relative strength measures demonstrated large to very large correlations with gross mechanical efficiency, 15-km time-trial velocity, maximum anaerobic power output, peak aerobic power output, power at a fixed blood lactate concentration of 4 mmol·L-1, and peak oxygen consumption (P = .05). CONCLUSION: Relative upper-body strength demonstrates a significant relationship with time-trial velocity and several handcycling performance measures. Relative strength is the product of one's ability to generate maximal forces relative to body mass. Therefore, the development of one's absolute strength combined with a reduction in body mass may influence real-world handcycling race performance.

The Anthropometric, Physiological, and Strength-Related Determinants of Handcycling 15-km Time-Trial Performance.

PURPOSE: The aim of this study was to investigate the relationship between selected anthropometric, physiological, and upper-body strength measures and 15-km handcycling time-trial (TT) performance. METHODS: Thirteen trained H3/H4 male handcyclists performed a 15-km TT, graded exercise test, 15-second all-out sprint, and 1-repetition-maximum assessment of bench press and prone bench pull strength. Relationship between all variables was assessed using a Pearson correlation coefficient matrix with mean TT velocity representing the principal performance outcome. RESULTS: Power at a fixed blood lactate concentration of 4 mmol·L-1 (r = .927; P < .01) showed an extremely large correlation with TT performance, whereas relative V˙O2peak (peak oxygen uptake) (r = .879; P < .01), power-to-mass ratio (r = .879; P < .01), peak aerobic power (r = .851; P < .01), gross mechanical efficiency (r = 733; P < .01), relative prone bench pull strength (r = .770; P = .03) relative bench press strength (r = .703; P = .11), and maximum anaerobic power (r = .678; P = .15) all demonstrated a very large correlation with performance outcomes. CONCLUSION: Findings of this study indicate that power at a fixed blood lactate concentration of 4 mmol·L-1, relative V˙O2peak, power-to-mass ratio, peak aerobic power, gross mechanical efficiency, relative upper-body strength, and maximum anaerobic power are all significant determinants of 15-km TT performance in H3/H4 handcyclists.

Update on the Role of Actovegin in Musculoskeletal Medicine: A Review of the Past 10 Years.

BACKGROUND: Actovegin is a biological drug with a controversial history of use in the treatment of sports injuries during the past 60 years. Particular concerns have been raised about its ergogenic potential to enhance performance, but some of these have been based on little more than anecdote. OBJECTIVES: In this article, we review the most recent scientific evidence to determine the clinical efficacy, safety profile, and legal status of Actovegin. METHODS: We considered all studies directly commenting on experience with Actovegin use as the primary intervention within the past 10 years. Outcomes included mechanisms of action, clinical efficacy in enhancing muscle repair, any report of safety issues, and any evidence for ergogenic effect. RESULTS: Our database search returned 212 articles, abstracts were screened, and after inclusion/exclusion criteria were applied, 25 articles were considered: Publications included 11 primary research articles (7 in vitro studies and 4 clinical trials), 8 review articles, 5 editorials, and a single case report. CONCLUSIONS: Current literature is still yet to define the active compound(s) of Actovegin, but suggests that it shows antioxidant and antiapoptotic properties, and may also upregulate macrophage responses central to muscle repair. Clinical efficacy was supported by one new original research article, and the use of Actovegin to treat muscle injuries remains safe and supported. Two articles argued the ergogenic effect of Actovegin, but in vitro findings did not to translate to the outcomes of a clinical trial. An adequate and meaningful scientific approach remains difficult in a field where there is immense pressure to deliver cutting-edge therapies.

Communities ready for takeoffIntegrating social assets for biofuel site-selection modeling.

Although much of the social science literature supports the importance of community assets for success in many policy areas, these assets are often overlooked when selecting communities for new infrastructure facilities. Extensive collaboration is crucial for the success of environmental and economic projects, yet it often is not adequately addressed when making siting decisions for new projects. This article develops a social asset framework that includes social, creative, and human capital to inform site-selection decisions. This framework is applied to the Northwest Advanced Renewables Alliance project to assess community suitability for biofuel-related developments. This framework is the first to take all necessary community assets into account, providing insight into successful site selection beyond current models. The framework not only serves as a model for future biorefinery projects but also guides tasks that depend on informed location selection for success.

Analysis of 17ß-estradiol, estriol and estrone in American eel (Anguilla rostrata) tissue samples using liquid chromatography coupled to electrospray differential ion mobility tandem mass spectrometry.

RATIONALE: 17ß-Estradiol (E2), estrone (E1) and estriol (E3) are steroid hormones responsible for the regulation of the female reproductive system. Estradiol is planned to be used to feminize eels in aquaculture in order to improve their size and marketability. The residual levels of these hormones in fish tissue must be monitored to meet the requirements of food regulatory agencies. Few studies have studied these hormones in complex biological matrices such as fish tissue. METHODS: We developed a method to analyze E1, E2 and E3 in fish tissue using liquid chromatography in combination with differential ion mobility spectrometry (DMS) and tandem mass spectrometry (MS/MS). The mass spectrometer was operated in negative polarity selected reaction monitoring (SRM) mode. To test the performance of this method, residual levels of E1, E2 and E3 were measured in the muscle tissue of juvenile eels subjected to feminization treatment with E2. RESULTS: We report that following 17ß-estradiol treatment, E2 is rapidly metabolized from the eel tissue, with a 50% depletion rate per day. Five days post-treatment, E2 returned to the level found in non-treated controls, similar to levels found in wild mature female eels. CONCLUSIONS: The method presented herein allows the quantitative analysis of E1, E2 and E3 in fish tissue samples. Under the experimental conditions, E2 in fish tissue samples returned to physiological levels post hormonal treatment. Copyright © 2017 John Wiley & Sons, Ltd.

Communities ready for takeoff.

Although much of the social science literature supports the importance of community assets for success in many policy areas, these assets are often overlooked when selecting communities for new infrastructure facilities. Extensive collaboration is crucial for the success of environmental and economic projects, yet it often is not adequately addressed when making siting decisions for new projects. This article develops a social asset framework that includes social, creative, and human capital to inform site-selection decisions. This framework is applied to the Northwest Advanced Renewables Alliance project to assess community suitability for biofuel-related developments. This framework is the first to take all necessary community assets into account, providing insight into successful site selection beyond current models. The framework not only serves as a model for future biorefinery projects but also guides tasks that depend on informed location selection for success.

The presence of highly disruptive 16S rRNA mutations in clinical samples indicates a wider role for mutations of the mitochondrial ribosome in human disease.

Mitochondrial DNA mutations are well recognized as an important cause of disease, with over two hundred variants in the protein encoding and mt-tRNA genes associated with human disorders. In contrast, the two genes encoding the mitochondrial rRNAs (mt-rRNAs) have been studied in far less detail. This is because establishing the pathogenicity of mt-rRNA mutations is a major diagnostic challenge. Only two disease causing mutations have been identified at these loci, both mapping to the small subunit (SSU). On the large subunit (LSU), however, the evidence for the presence of pathogenic LSU mt-rRNA changes is particularly sparse. We have previously expanded the list of deleterious SSU mt-rRNA mutations by identifying highly disruptive base changes capable of blocking the activity of the mitoribosomal SSU. To do this, we used a new methodology named heterologous inferential analysis (HIA). The recent arrival of near-atomic-resolution structures of the human mitoribosomal LSU, has enhanced the power of our approach by permitting the analysis of the corresponding sites of mutation within their natural structural context. Here, we have used these tools to determine whether LSU mt-rRNA mutations found in the context of human disease and/or ageing could disrupt the function of the mitoribosomal LSU. Our results clearly show that, much like the for SSU mt-rRNA, LSU mt-rRNAs mutations capable of compromising the function of the mitoribosomal LSU are indeed present in clinical samples. Thus, our work constitutes an important contribution to an emerging view of the mitoribosome as an important element in human health.

Influence of noncircular chainring on male physiological parameters in hand cycling.

The purpose of this study was to examine the influence of a noncircular chainring (NCC) compared with a conventional circular chainring (CC) on hand cycling performance. Eleven nondisabled male participants with no hand cycling experience initially completed an incremental exercise test. Afterward, the participants completed two 20 s sprint tests, followed by a 20 min endurance test and then another two 20 s sprint tests. An NCC and a CC were used in random order on two separate occasions. To compare the effects of the NCC and CC on power data of the sprint tests and metabolic response during the endurance test, a two-way analysis of variance for repeated measures was used. Average power values of the sprint tests showed no significant difference between NCC and CC, but over time, values of the first and third sprint tests were higher than those of the second and fourth sprint tests for both chainrings. Values of energy expenditure (kilojoules), gross efficiency (percentage), and net efficiency (percentage) after 10 and 20 min during the endurance test using NCC and CC showed no significant differences (p > 0.05) either between tests or over time. Under the current test conditions and focusing on physiological parameters, a performance optimization using an NCC in hand cycling could not be proven.

Heterologous inferential analysis (HIA) as a method to understand the role of mitochondrial rRNA mutations in pathogenesis.

Despite the identification of a large number of potentially pathogenic variants in the mitochondrially encoded rRNA (mt-rRNA) genes, we lack direct methods to firmly establish their pathogenicity. In the absence of such methods, we have devised an indirect approach named heterologous inferential analysis or HIA that can be used to make predictions on the disruptive potential of a large subset of mt-rRNA variants. First, due to the high evolutionary conservation of the rRNA fold, comparison of phylogenetically derived secondary structures of the human mt-rRNAs and those from model organisms allows the location of structurally equivalent residues. Second, visualization of the heterologous equivalent residue in high-resolution structures of the ribosome allows a preliminary structural characterization of the residue and its neighboring region. Third, an exhaustive search for biochemical and genetic information on the residue and its surrounding region is performed to understand their degree of involvement in ribosomal function. Additional rounds of visualization in biochemically relevant high-resolution structures will lead to the structural and functional characterization of the residue's role in ribosomal function and to an assessment of the disruptive potential of mutations at this position. Notably, in the case of certain mitochondrial variants for which sufficient information regarding their genetic and pathological manifestation is available; HIA data alone can be used to predict their pathogenicity. In other cases, HIA will serve to prioritize variants for additional investigation. In the context of a scoring system specifically designed for these variants, HIA could lead to a powerful diagnostic tool.

The role of the mitochondrial ribosome in human disease: searching for mutations in 12S mitochondrial rRNA with high disruptive potential.

Mutations of mitochondrial DNA are linked to many human diseases. Despite the identification of a large number of variants in the mitochondrially encoded rRNA (mt-rRNA) genes, the evidence supporting their pathogenicity is, at best, circumstantial. Establishing the pathogenicity of these variations is of major diagnostic importance. Here, we aim to estimate the disruptive effect of mt-rRNA variations on the function of the mitochondrial ribosome. In the absence of direct biochemical methods to study the effect of mt-rRNA variations, we relied on the universal conservation of the rRNA fold to infer their disruptive potential. Our method, named heterologous inferential analysis or HIA, combines conservational information with functional and structural data obtained from heterologous ribosomal sources. Thus, HIA's predictive power is superior to the traditional reliance on simple conservation indexes. By using HIA, we have been able to evaluate the disruptive potential for a subset of uncharacterized 12S mt-rRNA variations. Our analysis revealed the existence of variations in the rRNA component of the human mitoribosome with different degrees of disruptive power. In cases where sufficient information regarding the genetic and pathological manifestation of the mitochondrial phenotype is available, HIA data can be used to predict the pathogenicity of mt-rRNA mutations. In other cases, HIA analysis will allow the prioritization of variants for additional investigation. Eventually, HIA-inspired analysis of potentially pathogenic mt-rRNA variations, in the context of a scoring system specifically designed for these variants, could lead to a powerful diagnostic tool.

Altered 2-thiouridylation impairs mitochondrial translation in reversible infantile respiratory chain deficiency.

Childhood-onset mitochondrial encephalomyopathies are severe, relentlessly progressive conditions. However, reversible infantile respiratory chain deficiency (RIRCD), due to a homoplasmic mt-tRNA(Glu) mutation, and reversible infantile hepatopathy, due to tRNA 5-methylaminomethyl-2-thiouridylate methyltransferase (TRMU) deficiency, stand out by showing spontaneous recovery, and provide the key to treatments of potential broader relevance. Modification of mt-tRNA(Glu) is a possible functional link between these two conditions, since TRMU is responsible for 2-thiouridylation of mt-tRNA(Glu), mt-tRNA(Lys) and mt-tRNA(Gln). Here we show that down-regulation of TRMU in RIRCD impairs 2-thiouridylation and exacerbates the effect of the mt-tRNA(Glu) mutation by triggering a mitochondrial translation defect in vitro. Skeletal muscle of RIRCD patients in the symptomatic phase showed significantly reduced 2-thiouridylation. Supplementation with l-cysteine, which is required for optimal TRMU function, rescued respiratory chain enzyme activities in human cell lines of patients with RIRCD as well as deficient TRMU. Our results show that l-cysteine supplementation is a potential treatment for RIRCD and for TRMU deficiency, and is likely to have broader application for the growing group of intra-mitochondrial translation disorders.

Effects of playing surface on physiological responses and performance variables in a controlled football simulation.

In spite of the increased acceptance of artificial turf in football, few studies have investigated if matches are altered by the type of surface used and no research has compared physiological responses to football activity on artificial and natural surfaces. In the present study, participants performed a football match simulation on high-quality artificial and natural surfaces. Neither mean heart rate (171 ± 9 beats · min(-1) vs. 171 ± 9 beats · min(-1); P > 0.05) nor blood lactate (4.8 ± 1.6 mM vs. 5.3 ± 1.8 mM; P > 0.05) differed between the artificial and natural surface, respectively. Measures of sprint, jumping and agility performance declined through the match simulation but surface type did not affect the decrease in performance. For example, the fatigue index of repeated sprints did not differ (P > 0.05) between the artificial, (6.9 ± 2.1%) and natural surface (7.4 ± 2.4%). The ability to turn after sprinting was affected by surface type but this difference was dependent on the type of turn. Although there were small differences in the ability to perform certain movements between artificial and natural surfaces, the results suggest that fatigue and physiological responses to football activity do not differ markedly between surface-type using the high-quality pitches of the present study.

Immunoendocrine responses of male spinal cord injured athletes to 1-hour self-paced exercise: pilot study.

This study examined the effect of a 1 h, self-paced handcycling time trial on blood leukocytes, mucosal immunity, and markers of stress in paraplegic athletes. Nine male paraplegic athletes (spinal injury level thoracic 4-lumbar 2) performed 1 h of handcycling exercise on a standard 400 m athletics track. Heart rate (HR) was measured continuously during exercise, and a retrospective rating of perceived exertion (RPE) was obtained immediately after. Venous blood and saliva samples were collected immediately before exercise (Pre-Ex), after exercise (End-Ex), and 1 h postexercise (1-h Post). The athletes completed mean +/- standard error of mean 22.4 +/- 1.1 km cycling at HR 165 +/- 2 beats/min, RPE 15 +/- 1, and blood lactate 7.9 +/- 2.5 mmol/L. Total leukocytes increased 72% and neutrophils increased 74% End-Ex; both remained elevated at 1-h Post (both p < 0.05). Lymphocytes increased 53% and natural killer cells increased 175% End-Ex (both p < 0.05), but returned to near baseline levels 1-h Post. Increases (p < 0.05) were observed End-Ex in alpha-amylase activity (p < 0.05), which returned to baseline at 1-h Post, but there was no significant change in saliva flow rate, salivary immunoglobulin A, or cortisol. These data confirm that 1 h of handcycling exercise elevated circulating leukocytes but had a minimal effect on mucosal immunity. These changes appear to be associated with alpha-amylase rather than cortisol.

Altering the balance between healthy and mutated mitochondrial DNA.

Pathogenic mutations of the mitochondrial genome are frequently found to co-exist with wild-type mtDNA molecules, a state known as heteroplasmy. In most disease cases, the mutation is recessive with manifestation of a clinical phenotype occurring when the proportion of mutated mtDNA exceeds a high threshold. The concept of increasing the ratio of healthy to mutated mtDNA as a means to correcting the biochemical defect has received much attention. A number of strategies are highlighted in this article, including manipulation of the mitochondrial genome by antigenomic drugs or restriction endonucleases, zinc finger peptide-targeted nucleases and exercise-induced gene shifting. The feasibility of these approaches has been demonstrated in a number of models, however more work is necessary before use in human patients.