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It has been proposed that intermittent exercise can differentially affect mitochondrial responses to training, with training volume being more important than intensity for increasing skeletal muscle mitochondrial content and with intensity playing a greater role in mitochondrial respiration. To test this hypothesis, we examined markers of skeletal muscle mitochondrial content and respiration in response to two different interval training protocols performed using single-leg cycling, which permitted a within-subjects design. Ten healthy active adults [6 males / 4 females, 22±4 y, peak oxygen uptake (VO2peak) = 42±4 ml/kg/min] were recruited. Each leg was randomized to either a HIIT [4 × (5 min at 65% Wpeak and 2.5 min at 20% Wpeak)] or SIT [4 x (30-s “all-out” sprints and 4 min active recovery)] protocol and completed three exercise sessions/wk over 4 wk for a total of 12 exercise sessions/leg. The mean work performed during each session was 133±32 and 44±8.0 kJ for HIIT and SIT respectively, and the average workload during intervals was 95±25 W and 322±77 W for HIIT and SIT respectively. Citrate synthase (CS) maximal activity increased compared to baseline after training interventions, with the change being greater after SIT vs HIIT (42±25% vs 16±13%, interaction p=0.01). COXIV protein content and succinate-supported state 3 were unchanged. Single-leg VO2peak and time to exhaustion (TTE) increased to a similar extent in both HIIT and SIT (main effect of time, p<0.05). These data suggest that, in contrast to what has been proposed by others, training intensity is more important than volume for increasing mitochondrial content during short-term interval training in human skeletal muscle. / Thesis / Master of Science in Kinesiology / Mitochondria are an important component of cells that use oxygen to convert fuels such as sugars and fats into energy. One of the factors that determines the amount of mitochondria in skeletal muscle is physical activity. Aerobic exercise training can be performed over a range of intensities, from relatively easy to very hard, and in an intermittent or continuous manner. This thesis examined the effect of short-term, intermittent exercise training performed at two different intensities on the content of mitochondria in human skeletal muscle. It found that both high- intensity interval training (HIIT) and sprint interval training (SIT) increased mitochondrial content. The increase was greater after SIT compared to HIIT, even though the total “dose” or amount of exercise was lower in the former compared to the latter. These results suggest that intensity is an important determinant of skeletal muscle remodelling induced by intermittent exercise in humans.

This thesis considered three potential factors that may influence skeletal muscle responses to interval exercise training in healthy adults, with a focus on mitochondrial remodelling: biological sex, fibre type and contraction pattern. Study 1 assessed the acute response to a sprint interval training (SIT) protocol that involved three, 20-second ‘all-out’ cycling efforts interspersed with 2 minutes of recovery. It found similar exercise-induced increases in the mRNA expression of genes linked to mitochondrial biogenesis in men and women. Study 2 showed that a single session of SIT and moderate-intensity continuous training (MICT) induced similar increases in the phosphorylation of signalling proteins linked to mitochondrial biogenesis in type I and IIa fibres when compared within the same individuals. It also assessed responses to 12 weeks of training in two different groups and found MICT elicited greater increases in markers of mitochondrial content, including cytochrome c oxidase subunit IV (COXIV) protein content, in type I fibres, whereas the increase in type IIa fibres was similar compared to SIT. Study 3 utilized single-leg cycling as a within-subject model to compare interval and continuous training protocols that were matched for exercise intensity and total training volume, but the pattern of contraction differed. Measurements of mitochondrial content, including subsarcolemmal mitochondrial area, the maximal activity of citrate synthase and COXIV protein content, increased after 4 weeks of continuous training but were unchanged after interval training. Overall, this thesis advances our understanding of the influence of biological sex, muscle fibre type and contraction pattern on skeletal muscle mitochondrial responses to exercise. Major findings include: the acute response to SIT was similar between sexes, acute responses to interval and continuous exercise were similar but training elicited some divergent fibre type-specific responses, and mitochondrial content increased following continuous training but was unchanged following work- and intensity-matched interval training. / Dissertation / Doctor of Philosophy (PhD) / This thesis considered the role of biological sex, fibre type and contraction pattern on skeletal muscle responses to exercise. It focused on mitochondria, which are important components of the cell that affect health and performance. The responses of genes that regulate the formation of new mitochondria to a single session of brief, “sprint” interval exercise were similar between men and women. Acute sprint interval exercise also activated molecular pathways similar to traditional endurance exercise in the two main types of muscle fibres; however, 12 weeks of endurance training induced greater increases in mitochondrial content in type I (“slow-twitch”) fibres, whereas the increases in type II (“fast-twitch”) fibres were similar compared to interval training. Lastly, mitochondrial content increased after continuous training but not interval training when the intensity and total amount of exercise was matched. Our findings improve our understanding of the mechanisms by which exercise elicits mitochondrial remodelling in humans.

Skeletal muscle satellite cells (SC) are essential in muscle repair and regeneration. The role of SCs in mediating hypertrophic adaptations following resistance training has been widely studied. Recent evidence from endurance training studies suggest that SCs may also play a role in mediating non-hypertrophic adaptations. Indeed, it has been shown that satellite cells respond to endurance training. Work in rodent models suggest that exercise intensity may play an important role in expanding the SC pool whereas the results of endurance training studies in humans are much less consistent. Limited evidence also suggest that exercise intensity may be important in mediating exercised-induced capillarization following endurance training in humans. In both instances, it is unknown whether the on-and-off pattern characteristic of interval training (i.e. the rest-work cycles) plays a role in the magnitude of these skeletal muscle responses to this type of exercise. Thus, we sought to determine if the rest-to-work cycle plays a role in the scope of these skeletal muscle responses by comparing the SC response and capillarization to two distinct work-matched protocols that are performed at the same intensity but at two different exercise patterns (interval versus continuous). We hypothesized that interval exercise will elicit a greater SC response and induce greater capillary growth compared to work-matched continuous exercise. Ten young active individuals performed 12 sessions of counterweighted single-leg cycling over 4 weeks. Each leg was randomly assigned to Interval (INT) (10 x 3-min intervals at 50% of single leg peak power output (PPO), with 1 min recovery) or Continuous (CONTIN) (30 min at 50% PPO, followed by 10 min recovery), which were performed 5 min apart on each day, in an alternating order. Resting muscle biopsies were obtained from the vastus lateralis pre- and post-training. Immunofluorescent microscopy of muscle cross sections was used to assess muscle fibre capillarization, SC expansion and activation. Western blot analysis was performed on pro-angiogenic factors, specifically VEGF and VEGFR2. Mixed muscle SC activation increased in the INT leg compared to the pre-training time-point (3.2 ± 0.5 vs. 1.5 ± 0.2 Pax7+/MyoD+ cells/ 100 myofibers, respectively; P < 0.05). Mixed muscle SC activation did not increase significantly in the CONTIN leg compared to the pre-training time-point and there was no significant difference in activation between CONTIN and INT post-training. With regards to capillarization, no differences in type I or type II muscle fibre CC (capillary contacts), C/Fi (individual capillary-to-fibre ratio) or CFPE (capillary-to-fibre perimeter exchange index) were observed post-training or between legs. No significant expansion of the SC pool occurred in either legs post-training and no increases in fibre cross-sectional area was observed. This study presents novel evidence of mixed muscle SC activation following interval exercise training that is not observed following work- and intensity-matched continuous exercise training. This suggest that the rest-to-work cycle associated with interval exercise may dictate, to some extent, SC activation whereas it may not be a primary stimulus for training-induced changes in capillarization. / Thesis / Master of Science (MSc)

Studies were conducted on the effects of alcohol and opioids on the development of forensically important flies. In addition different methods of degree-day calculations and development thresholds were used to determine the effects on PMI estimates. The first study determined the effects of ethanol on the development of Phormia regina in vitro. Ground pork loin was treated with a 1, 5, or 10% ethanol solution to give an equivalent Blood Alcohol Concentration (BAC) of 0.01, 0.04, and 0.8 % w/v. A significant difference in the time for second instars to complete the stage was seen between the 1% treated and control. Significant differences were also found among pupal and adult weights between all treatments and the control. A significant difference was shown between growth curves of the 5% treated and control for third instar larvae using the Kolmogorov-Smirnov test. The ethanol content of Phormia regina in migrating third instar larvae that fed on treated meat was examined using headspace-gas chromatography (HSGC). All larvae had a content of 0.01% w/v, including the control. The effects of ante-mortem injection of oxycodone in pigs were examined with respect to insect succession patterns and the development rates of Phormia regina. Pigs were given a subcutaneous injection of oxycodone hydrochloride (3 mg/kg by weight) and antemortem blood samples were collected prior to and following drug injection. Shortly after death the carcasses were placed at an open field site and allowed to decompose in a field cage. Insect samples were collected from carcasses for seven days post-mortem and the collected data were used to develop occurrence matrices. The Simple Matching Coefficient showed that successional patterns were similar between treated and untreated animals. Loin and liver from the carcasses were used as rearing media for in vitro development studies of Phormia regina. Kolmogorov-Smirnov test showed that third instar P. regina maggots from treated loin tissue were significantly longer in length than maggots feeding on untreated loin tissue. A significant difference in time was found among larvae on loin for the time from eclosion to completion of the second instar. Significant differences were seen in the weight of adults reared on liver and loin. A chi-square for homogeneity showed that adults were biased towards males (2:1) from untreated loin tissue. A final study compared weather data sources, Accumulated Degree-Day (ADD) methods, and postmortem interval (PMI) estimations based on threshold and developmental data source. Four pigs were used for statistical comparisons. Pigs were taken to a test site and allowed to decompose in an experimental cage. Probes recorded ambient temperatures and body temperatures. Maggot sampling was completed every day for each pig. A three way factorial linear fit model was used to test for statistical differences. Significant differences were seen in the calculated ADD based on probe location and the development threshold used. The ADD calculated from local weather station locations: Kentland Farm, Moore Farm, and Blacksburg Airport were also compared. A significant difference in ADD was found in the main effects among locations (Airport 44.1 ADD, Kentland 37.5 ADD, Moore 48.6 ADD), as well as among the thresholds used (10,12.2, and 14°C). Different PMI estimations also resulted when using development data from different development studies on Phormia regina. / Ph. D.

This work is focused on overdetermined systems of interval linear equati- ons. First part consists of introduction to interval arithmetics and interval linear algebra and basic theory of interval linear systems. In the second part various methods for solving overdetermined interval linear systems are de- scribed. By solution of overdetermined interval system we mean union of all solutions of all subsystems. Known and our variants of algorithms are discussed. We introduce our subsquare method. All mentioned methods are implemented in one toolbox for Matlab. Methods are tested on solvable and unsolvable overdetermined systems. For solvable systems we test solution enclosure, time and special features of methods. For unsolvable systems we test detection of unsolvability. At the end of this work we provide basic in- troduction to Intlab. 1

Despite clear recommendations on the minimal amount of physical activity for achieving health benefits and reducing risk of chronic disease, the majority of people in the Western world remain sedentary. As a 'lack of time' has been identified as one of the main barriers to becoming and remaining physically active, in the past decade research has focused on high-intensity interval training (HIT) as a time-efficient alternative to aerobic exercise. Although initial studies convincingly demonstrated equal or better health benefits with various HIT protocols compared to much larger volumes of aerobic exercise, these HIT protocols tend to be very strenuous and as such are unlikely to be adhered to by sedentary populations. Furthermore, most HIT protocols are not as time-efficient as sometimes claimed, with the total time per exercise session generally exceeding 20-30 minutes. This thesis aimed to characterise the effects of a novel reduced-exertion HIT (ReHIT) protocol, requiring a maximum of 2 x 20 s all-out sprint efforts in a 10 min training session, upon insulin sensitivity, aerobic capacity, glycogen utilisation and associated acute metabolic responses. The ReHIT exercise bouts were well tolerated by participants, but were associated with a substantial disturbance of physiological homeostasis including muscle glycogen degradation, lactate accumulation, excursions in plasma volume, post-exercise oxygen consumption, respiratory exchange ratio and heart rate, as well as a skeletal muscle signalling response through AMPK, and increases in skeletal muscle GLUT4 and PGC1α mRNA expression (Chapter 4 and 5). The combined training studies (n=49) provide some support for improvements in key disease biomarkers following ReHIT, with improvements in insulin sensitivity observed in men, and increased aerobic capacity observed in men and women (Chapter 7). These observations highlight a potential regulatory role for glycogen in exercise-induced adaptation. However, the mean improvements in insulin sensitivity in men were not consistent between the two training studies (Chapter 2 and 6), and there was a high level of variability observed between individuals (Chapter 6 and 7). Therefore, the impact of ReHIT on insulin sensitivity needs to be further explored in the context of a randomised controlled trial, and the mechanisms underpinning the large variability in adaptive response need to be characterised.

Consideramos um modelo de regressão linear simples, em que tanto a variável resposta como a independente estão sujeitas a censura intervalar. Como motivação utilizamos um estudo em que o objetivo é avaliar a possibilidade de previsão dos resultados de um exame audiológico comportamental a partir dos resultados de um exame audiológico eletrofisiológico. Calculamos intervalos de previsão para a variável resposta, analisamos o comportamento dos estimadores de máxima verossimilhança obtidos sob o modelo proposto e comparamos seu desempenho com aquele de estimadores obtidos de um modelo de regressão linear simples usual, no qual a censura dos dados é desconsiderada. / We consider a simple linear regression model in which both variables are interval censored. To motivate the problem we use data from an audiometric study designed to evaluate the possibility of prediction of behavioral thresholds from physiological thresholds. We develop prediction intervals for the response variable, obtain the maximum likelihood estimators of the proposed model and compare their performance with that of estimators obtained under ordinary linear regression models.

Comparative experiments, in which subjects are randomized to one of two treatments, are performed often. There is no shortage of papers testing whether a treatment effect exists and providing confidence intervals for the magnitude of this effect. While it is well understood that the object and scope of inference for an experiment will depend on what assumptions are made, these entities are not always clearly presented. We have proposed one possible method, which is based on the ideas of Jerzy Neyman, that can be used for constructing confidence intervals in a comparative experiment. The resulting intervals, referred to as Neyman-type confidence intervals, can be applied in a wide range of cases. Special care is taken to note which assumptions are made and what object and scope of inference are being investigated. We have presented a notation that highlights which parts of a problem are being treated as random. This helps ensure the focus on the appropriate scope of inference. The Neyman-type confidence intervals are compared to possible alternatives in two different inference settings: one in which inference is made about the units in the sample and one in which inference is made about units in a fixed population. A third inference setting, one in which inference is made about a process distribution, is also discussed. It is stressed that certain assumptions underlying this third type of inference are unverifiable. When these assumptions are not met, the resulting confidence intervals may cover their intended target well below the desired rate. Through simulation, we demonstrate that the Neyman-type intervals have good coverage properties when inference is being made about a sample or a population. In some cases the alternative intervals are much wider than necessary on average. Therefore, we recommend that researchers consider using our Neyman-type confidence intervals when carrying out inference about a sample or a population as it may provide them with more precise intervals that still cover at the desired rate.