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.

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.

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.

A functional peptidyl-tRNA hydrolase, ICT1, has been recruited into the human mitochondrial ribosome.

Bioinformatic analysis classifies the human protein encoded by immature colon carcinoma transcript-1 (ICT1) as one of a family of four putative mitochondrial translation release factors. However, this has not been supported by any experimental evidence. As only a single member of this family, mtRF1a, is required to terminate the synthesis of all 13 mitochondrially encoded polypeptides, the true physiological function of ICT1 was unclear. Here, we report that ICT1 is an essential mitochondrial protein, but unlike the other family members that are matrix-soluble, ICT1 has become an integral component of the human mitoribosome. Release-factor assays show that although ICT1 has retained its ribosome-dependent PTH activity, this is codon-independent; consistent with its loss of both domains that promote codon recognition in class-I release factors. Mutation of the GGQ domain common to ribosome-dependent PTHs causes a loss of activity in vitro and, crucially, a loss of cell viability, in vivo. We suggest that ICT1 may be essential for hydrolysis of prematurely terminated peptidyl-tRNA moieties in stalled mitoribosomes.

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.

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.

Nuclear factors involved in mitochondrial translation cause a subgroup of combined respiratory chain deficiency.

Mutations in several mitochondrial DNA and nuclear genes involved in mitochondrial protein synthesis have recently been reported in combined respiratory chain deficiency, indicating a generalized defect in mitochondrial translation. However, the number of patients with pathogenic mutations is small, implying that nuclear defects of mitochondrial translation are either underdiagnosed or intrauterine lethal. No comprehensive studies have been reported on large cohorts of patients with combined respiratory chain deficiency addressing the role of nuclear genes affecting mitochondrial protein synthesis to date. We investigated a cohort of 52 patients with combined respiratory chain deficiency without causative mitochondrial DNA mutations, rearrangements or depletion, to determine whether a defect in mitochondrial translation defines the pathomechanism of their clinical disease. We followed a combined approach of sequencing known nuclear genes involved in mitochondrial protein synthesis (EFG1, EFTu, EFTs, MRPS16, TRMU), as well as performing in vitro functional studies in 22 patient cell lines. The majority of our patients were children (<15 years), with an early onset of symptoms <1 year of age (65%). The most frequent clinical presentation was mitochondrial encephalomyopathy (63%); however, a number of patients showed cardiomyopathy (33%), isolated myopathy (15%) or hepatopathy (13%). Genomic sequencing revealed compound heterozygous mutations in the mitochondrial transfer ribonucleic acid modifying factor (TRMU) in a single patient only, presenting with early onset, reversible liver disease. No pathogenic mutation was detected in any of the remaining 51 patients in the other genes analysed. In vivo labelling of mitochondrial polypeptides in 22 patient cell lines showed overall (three patients) or selective (four patients) defects of mitochondrial translation. Immunoblotting for mitochondrial proteins revealed decreased steady state levels of proteins in some patients, but normal or increased levels in others, indicating a possible compensatory mechanism. In summary, candidate gene sequencing in this group of patients has a very low detection rate (1/52), although in vivo labelling of mitochondrial translation in 22 patient cell lines indicate that a nuclear defect affecting mitochondrial protein synthesis is responsible for about one-third of combined respiratory chain deficiencies (7/22). In the remaining patients, the impaired respiratory chain activity is most likely the consequence of several different events downstream of mitochondrial translation. Clinical classification of patients with biochemical analysis, genetic testing and, more importantly, in vivo labelling and immunoblotting of mitochondrial proteins show incoherent results, but a systematic review of these data in more patients may reveal underlying mechanisms, and facilitate the identification of novel factors involved in combined respiratory chain deficiency.

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.

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.

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.

Acute liver failure with subsequent cirrhosis as the primary manifestation of TRMU mutations.

Combined respiratory chain deficiency accounts for about 30% of mitochondrial respiratory chain deficiencies and is frequently associated with mtDNA depletion, deletions or point mutations. However combined respiratory chain deficiency may also be caused by mutations in nuclear genes affecting mitochondrial translation. Here we describe a 2-year-old girl, who developed an acute, isolated, severe liver failure with mitochondrial pathology and decreased respiratory chain enzyme activities both in liver and skeletal muscle at 4 months of age. Her liver function improved significantly within a month, liver function tests returned to normal. Liver cirrhosis remained without any further complications so far. Pathogenic compound heterozygous mutations were identified in the TRMU gene. This condition is one of the few mitochondrial disorders with a life-threatening onset showing recovery later in life, therefore a prompt diagnosis and treatment of these patients has great importance in clinical practice. We suggest that TRMU deficiency should be considered in infants with acute liver disease.

The effects of an increasing versus constant crank rate on peak physiological responses during incremental arm crank ergometry.

We examined the effects of concomitant increases in crank rate and power output on incremental arm crank ergometry. Ten healthy males undertook three incremental upper body exercise tests to volitional exhaustion. The first test determined peak minute power. The subsequent tests involved arm cranking at an initial workload of 40% peak minute power with further increases of 10% peak minute power every 2 min. One involved a constant crank rate of 70 rev · min(-1), the other an initial crank rate of 50 rev · min(-1) increasing by 10 rev · min(-1) every 2 min. Fingertip capillary blood samples were analysed for blood lactate at rest and exhaustion. Local (working muscles) and cardiorespiratory ratings of perceived exertion (RPE) were recorded at the end of each exercise stage. Heart rate and expired gas were monitored continuously. No differences were observed in peak physiological responses or peak minute power achieved during either protocol. Blood lactate concentration tended to be greater for the constant crank rate protocol (P = 0.06). Test duration was shorter for the increasing than for the constant crank rate protocol. The relationship between local RPE and heart rate differed between tests. The results of this study show that increasing cadence during incremental arm crank ergometry provides a valid assessment of peak responses over a shorter duration but alters the heart rate-local RPE relationship.

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.

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.

Molecular basis of infantile reversible cytochrome c oxidase deficiency myopathy.

Childhood-onset mitochondrial encephalomyopathies are usually severe, relentlessly progressive conditions that have a fatal outcome. However, a puzzling infantile disorder, long known as 'benign cytochrome c oxidase deficiency myopathy' is an exception because it shows spontaneous recovery if infants survive the first months of life. Current investigations cannot distinguish those with a good prognosis from those with terminal disease, making it very difficult to decide when to continue intensive supportive care. Here we define the principal molecular basis of the disorder by identifying a maternally inherited, homoplasmic m.14674T>C mt-tRNA(Glu) mutation in 17 patients from 12 families. Our results provide functional evidence for the pathogenicity of the mutation and show that tissue-specific mechanisms downstream of tRNA(Glu) may explain the spontaneous recovery. This study provides the rationale for a simple genetic test to identify infants with mitochondrial myopathy and good prognosis.

The effects of a respiratory warm-up on the physical capacity and ventilatory response in paraplegic individuals.

A respiratory warm-up (RWU) can improve exercise performance in able-bodied athletes. However, its effects in paraplegic individuals are unknown. On two occasions, nine male active paraplegic individuals performed an arm cranking test to exhaustion at 85% of their peak power output. In the intervention (INT) trial, this procedure was preceded by a RWU, whereas in the control (CON) trial, no RWU was conducted. Time to exhaustion was reduced following the RWU (CON vs. INT: 497 ± 163 vs. 425 ± 126 s, P = 0.02). Pulmonary ventilation was increased in the middle (74.8 ± 18.0 vs. 78.3 ± 19.6 L min(-1), P = 0.01) and end (86.1 ± 20.4 vs. 95.4 ± 23.3 L min(-1), P = 0.01) phase of exercise following the RWU. Forced expiratory volume in 1 s (FEV1) was reduced following the RWU (3.44 ± 0.45 vs. 3.27 ± 0.54 L, P = 0.02). The decrease in FEV1 following the RWU and the higher pulmonary ventilation during the INT trial suggest that the RWU fatigued the respiratory system, and hence reduced performance capacity. It is possible that the RWU used in this study is not suitable for paraplegic individuals, as their respiratory system is limited due to their disability. We conclude that a RWU impaired exercise performance in a group of active paraplegic individuals as a result of respiratory muscle fatigue.