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No / The biochemistry of hair pigmentation is a complex field involving a plethora of protein and peptide mechanisms. The in loco factory for melanin formation is the hair follicle melanocyte, but it is common knowledge that melanogenesis results from a fine tuned concerted interaction between the cells of the entire dermal papilla in the anagen hair follicle. The key enzyme is tyrosinase to initiate the active pigmentation machinery. Hence, an intricate understanding from transcription of mRNA to enzyme activity, including enzyme kinetics, substrate supply, optimal pH, cAMP signaling, is a must. Moreover, the role of reactive oxygen species on enzyme regulation and functionality needs to be taken into account. So far our knowledge on the entire hair cycle relies on the murine model of the C57BL/6 mouse. Whether this data can be translated into humans still needs to be shown. This article aims to focus on the effect of H2O2-redox homeostasis on hair follicle pigmentation via tyrosinase, its substrate supply and signal transduction as well as the role of methionine sulfoxide repair via methionine sulfoxide reductases A and B (MSRA and B).

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Intense exercise is known to cause alterations in the psychophysiological status of an athlete. Monitoring the health and recovery of an athlete is imperative for the maintenance of performance and reduced fatigue and injury incidence. The physicality associated with select sports results in significant elevations and suppression of psychophysiological biomarkers that are often modulated by game-related impacts, intense training regimes and psychosocial aspects associated with the professional era. The aim of the studies outlined in this thesis were to determine the effectiveness of selected “stress” markers in several sports that result in significant “stress”, and quantify the level of acute and chronic “stress” following individual games and competitions to improve athlete management and recovery. Study one aimed at developing a new strong-cation exchange high performance liquid chromatography (SCX-HPLC) method for the detection and quantification of urinary pterins and creatinine in a body-building cohort completing high intensity resistance training. The method had an intra- and inter-assay variability of 3.04 % and 5.42 % respectively, with visibly clear peaks and no tailing. Urinary neopterin (NP) and 7,8-dihydroneopterin during a week of competitive natural body-building did not significantly change indicating no alteration in immune system function and oxidative stress. It did provide evidence for the use of specific gravity as a similarly reliable method for urine volume correction following exercise. Study two focused on a playoff game of elite amateur rugby. The time course changes of NP, cortisol, salivary immunoglobulin A (sIgA) and myoglobin in 11 elite amateur rugby players were measured up to 86 hours post-game. Cortisol increased 4-fold, myoglobin 2.85-fold, NP 1.75-fold and total NP 2.3-fold, all significant, whilst sIgA did not change. All markers returned to baseline within 17 hours providing valuable information for sample collection schedule optimization. Respiratory elastance was also measured by ventilation for the assessment of exercise induced lung inflammation/injury following the game (Chapter three). There was an increase in elastance in selected individuals that did not correlate with either global positioning system (GPS) or impact data. It was shown however, that a ventilator is capable of measuring respiratory changes in a conscious and healthy individual. Study three focused on the final three games of professional rugby in the 2013 ITM Cup. The acute and cumulative changes in the same four markers were analysed following three home games. There were significant increases in NP, total NP, cortisol and myoglobin along with significant suppression of sIgA (p < 0.05). Large intra- and inter-individual variation existed between players with changes associated with total impacts. Moreover, impact induced muscle damage may account for changes in oxidative status. Specific gravity (SG) was shown to be a more reliable marker for urine volume correction in comparison to creatinine; while some players showed signs of cumulative stress. Study four examined stress in a professional team throughout the 22 week 2014 Super 15 competition. Part one investigated changes in oxidative stress and muscle damage markers to solidify the muscle damage/oxidative status theory postulated in the previous study. Experimental evidence showed iron and myoglobin are separately capable of oxidizing 7,8-dihydroneopterin to NP in vitro. It was then identified that players who suffered the greatest muscle damage as a result of impacts also had the greatest change in oxidative status (NP). This evidence suggests rugby union induces significant alterations in oxidative status that may be exacerbated by the impact induced release of myoglobin. Part two measured urinary NT-proBNP during the last two consecutive home games to identify whether rugby union causes significant cardiovascular stress and if the pre to post-game change can be explained by GPS technology. Significant individualized elevations were observed in games one and two which did not correlate with any GPS measurements or impacts. Concentrations returned to normal ~ 36 hours post-game suggesting no permanent damage to cardiac muscle had occurred. The lack of correlation suggests GPS technology is not an accurate measure of cardiovascular stress in professional rugby union. Part three involved the measurement of cortisol, total NP and sIgA throughout the season to assess the degree of cumulative stress. Samples were taken at regular intervals ~ 36 hours post-game for 22 weeks. Extreme inter-individual variation was present. Select individuals showed continual elevation in immune system activation and psychophysiological stress, whilst others presented with a continual decline in immune system function. Collectively however, minor deviations from baseline in all markers were observed and participation in long distance travel did not significantly affect the psychophysiological status of the group. Together this suggests a season does not cause an accumulation in psychophysiological stress, although careful individual player analysis is warranted. Understanding rugby union positional demands is essential for training program specification and position specific development of players. Part four used GPS, video-analysis and biochemical analysis to identify positional demands in five regular season games. Forwards tended to be involved in more impacts and covered less distance, while backs covered more distance and carried the ball into contact more regularly. There was no difference in the psychophysiological status between positions indicating both aspects of stress (impacts and distance covered) may induce a similar response. Alternatively, individual biological variation may be solely responsible for this change suggesting careful consideration should be given when using traditional work-load measures such as GPS when quantifying “stress”. Part five assessed the effectiveness of varied recovery interventions. Total NP, cortisol, myoglobin and sIgA were measured pre- post- and ~ 36 hours post game to identify which intervention was most effective at returning players to a psychophysiological state that allowed for the resumption of normal training. Findings concluded the immediate post-game strategy employed by the team (cold bath, consumption of protein and carbohydrates, compression garments and eight hours sleep) seemed to provide the greatest psychophysiological improvement regardless of the “next-day” intervention. There was large inter-individual variation and players were still in a state of recovery ~ 36 hours post-game as indicated by the elevated total NP and sIgA concentrations. Study five had four aspects. Develop a new, cost-effective and simple reverse phase HPLC (RP-HPLC) method for the quantification of urinary myoglobin in a clinically relevant range, quantify the level of structural stress following a simulated mixed martial arts (MMA) contest, determine whether cold water immersion attenuates the level of inflammation and muscle damage following a contest, and whether this hypothesized attenuation may be explained by cryotherapy induced mononuclear cell activation suppression in vitro. The RP-HPLC method had an intra- and inter-assay variations from 0.32 - 2.94 %. Linearity was in the range of 5 – 1000 µg/mL which detected significant increases in urinary myoglobin following the MMA contest. Total NP was found to significantly increase following the contest and return to approximately pre-contest levels 24 hours later for the passive group only. Cold water immersion was further found to attenuate the total NP increase in the first two hours post-contest solidifying its use as a recovery technique following intense exercise, while cryotherapy significantly suppressed T-cell activation. This study provides a reliable and repeatable assay for muscle damage quantification in a clinically relevant range, evidence of the physicality associated with MMA, and indicates cold water immersion is a reliable recovery intervention that may impart its positive benefits through T-cell suppression. The data generated by these investigations highlights the necessity for individual physiological analysis. Group data often masks the extreme variation that exists in clinical and exercise trials where treatment and management of athletes is conducted for recovery and performance. Biochemical analysis provides an added sophistication of work-load and psychophysiological assessment that common technological methods cannot emulate. With a lack of correlation between the quantitative changes in specific non-overlapping biomarkers and GPS, video-analysis and questionnaires, it would seem pertinent to develop a non-invasive quantitative approach in elite sport to understand the level of exercise-induced psychophysiological stress for the precise management of athletes.