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No description available.

Accurate and reliable measurement of meat quality is essential for the Australian beef industry to remain competitive in both the domestic and export markets. Recent developments of the resonator technique have lead to the commercial availability of the High Resolution Ultrasound Spectroscope (HR-US). This research project was designed to assess the potential of HR-US for the analysis of post-mortem bovine muscle. This was accomplished by; 1) establishing a suitable measurement protocol that considered sources of variability, 2) the effects of post-mortem aging on HR-US parameters, 3) analysis of thermal related changes that occur in muscle, and extracted connective tissue during heating, and 4) the use of HR-US for the measurement of the intramuscular fat. A procedure for the measurement of bovine muscle with HR-US was established. Briefly, an external semisolid cell was used as the measurement cell. The frequency range of 2000 KHz to 3000 KHz was selected as the most suitable for whole muscle analysis and all five resonance peaks within this range were analysed and used to obtain velocity and attenuation values of the meat sample. Water was used as the reference media, and measurements were conducted at 250C. Changes were made to this method during experimental work depending on the sample being run and the objectives of the study. The measurement protocol was shown to be repeatable. Factors likely to cause variation in measurements of the samples, such as water loss and freeze-thaw, were also considered when developing the operational parameters of the study. High resolution ultrasound spectroscopy was applied to measure the post-mortem changes that occur in bovine muscle. Using two muscle types, Semitendinosus and Psoas major, significant changes were observed in HR-US parameters with ageing. Significant increases in the acoustic impedance of bovine muscle with increased ageing time were attributed to degradation of the muscle structure. This was confirmed in transmission electron microscopy images where clear disruption the myofibillar structure was apparent in the muscle at 21 days post-mortem.In localised regions the Z bands and the adjoining actin fibres were totally degraded. Water loss from the muscle system had a significant influence on HR-US measurements. Thermal related changes that occur in whole bovine muscle and in isolated intramuscular connective tissue were observed with HR-US. Heat induced changes were identified in whole muscles and included the melting of the fat within samples at 48oC, coagulation of sarcoplasmic proteins between 450C and 55oC, and the shrinkage of collagen fibres at 630C. An 80% reduction in the attenuating properties of extracted connective ageing in buffer solution was observed within the first 5 days. This is attributed to the degradation of proteoglycans and the resulting disassociation of collagen fibrils. Structural changes occurring in extracted connective tissue were observed with TEM. HR-US measurements proved to be highly sensitive to identifying temperatures at which transitions occurred. Unfolding of the triple helix structure of collagen was identified in velocity transitions between 59°C and 63oC. HR-US results suggested a greater sensitivity to thermal related changes in extracted intramuscular connective tissue when compared with differential scanning calorimeter results. An increase in temperature was observed for thermal denaturation of collagen with ageing, however a reduction was also observed in the temperature range at which the denaturation process occurred. Temperature ramps conducted on extracted intramuscular bovine showed a reduction in velocity from 1613.1m/s at 250C to 1343.1 m/s at 900C equalling an overall reduction in velocity of 270m/s. A transition in the velocity trend seen at 46°C indicates the majority of the triglycerides are melted (or in liquid state) above this temperature. Results are confirmed with differential scanning calorimeter thermogram. HR-US measurements showed high sensitivity to increasing concentration of bovine fat in prepared emulsions with an adjusted R2 99.46% for velocity measurements taken at 5100 kHz. Attenuation values at 8100 kHz also showed a strong linear response to increasing fat concentration in the emulsion (R2 98.77). The use of HR-US for the measurement of intramuscular bovine fat demonstrated a high sensitivity to extracted bovine fat in prepared emulsions. An increase in the intramuscular fat content of whole bovine muscles resulted in a reduction in the velocity measurements and an increase in the attenuation of the ultrasonic signal. This provides the basis for potential method for the prediction intramuscular fat in bovine muscle. The present studies have highlighted the complexities of investigations relating to meat quality and have demonstrated the diversity of data required to assess quality. Only when comprehensive data are available, can we hope to accurately determine meat quality and predict how it will vary with changes in animal production and meat processing.

Diss. (sammanfattning) Umeå : Umeå universitet, 2007. / Härtill 5 uppsatser.

Le maintien ou le renforcement de la masse et de la fonction musculaire, altérées chez les patients BPCO, est un objectif primordial pour préserver, voire améliorer leur tolérance à l'effort, leur qualité de vie et leur survie. Afin d'optimiser la prise en charge de cette dysfonction musculaire, la réhabilitation est complétée par des interventions nutritionnelles, encore appelées réhabilitations nutritionnelles. Dans ce contexte, l'apport d'acides gras polyinsaturés de la série n-3, et plus particulièrement d'acide docosahexaénoïque (DHA), pourrait s'avérer intéressant en raison de leurs effets bénéfiques démontrés dans plusieurs pathologies chroniques. L'objectif de ce travail était donc de caractériser les effets d'une supplémentation en DHA sur la tolérance à l'effort et sur le métabolisme énergétique des muscles squelettiques de rats adultes exposés à une hypoxie comme modèle de muscle de patient BPCO au stade de l'insuffisance respiratoire chronique. La tolérance à l'effort est améliorée par le DHA, que les rats soient conditionnés en normoxie ou en hypoxie. En normoxie, les mécanismes impliqués seraient liés à un effet du DHA mimétique de celui d'un exercice d'endurance, avec une activation de l'AMPK et une amélioration de la fonction mitochondriale étudiée sur fibres musculaires perméabilisées. En hypoxie, le DHA agirait différemment, réduisant les effets de l'hypoxie sur le muscle, sans que les mécanismes mimétiques de l'exercice d'endurance ne soient clairement retrouvés. La prise de DHA chez des rats entrainés et conditionnés en hypoxie permet également un gain d'endurance mais les mécanismes à l'origine de cet effet ne sont pas élucidés et nécessitent des travaux complémentaires. Au vu des résultats sur le muscle, la supplémentation en DHA pourrait donc être bénéfique dans la prise en charge de la dysfonction musculaire dans les maladies chroniques telles que la BPCO. / The maintenance and reinforcement of skeletal muscle mass and function, impaired in COPD patients, is a crucial aim to preserve, and even improve, their exercise tolerance, quality of life and survival. In order to optimize the management of such a muscular dysfunction, rehabilitation could be completed by nutritional interventions, also called nutritional rehabilitation. In this context, the intake of polyunsaturated fatty acids of the n-3 class, and particularly of docosahexaenoic acid (DHA), could be interesting, due to their benefic effects demonstrated in many chronic pathologies. Therefore, the aim of our work was to characterize the effects of DHA supplementation on exercise tolerance and skeletal muscle metabolism in rats exposed to hypoxia as a model of muscle dysfunction as seen in COPD patients suffering chronic respiratory failure. Results showed that exercise tolerance was improved by DHA in both normoxia and hypoxia conditions. In normoxia, the involved mechanisms may rely on an endurance exercise mimetic effect of DHA, including AMPK activation and improved mitochondrial function studied on permeabilized muscular fibers. In hypoxia, DHA acts differently, probably by minimizing hypoxia effects on muscle. However, the endurance exercise mimetic mechanisms were not clearly found. In hypoxia exposed-endurance trained-rats, DHA improved endurance exercise capacity but the involved mechanisms were not fully characterized and need further work. In conclusion, our results on muscle suggest that DHA supplementation could be beneficial in management of muscular dysfunction induced by chronic diseases such as COPD.

Abstract The function of skeletal muscle is to generate force and produce movement. These tasks are carried out by long multinucleated cells, the skeletal myofibres. The membrane system and the cytoskeleton of these cells are uniquely organized to respond rapidly to neuronal stimuli and to achieve efficient contraction. In the present study the organization and distribution of selected protein/lipid based microdomains that reside in the plasma membrane and sarcoplasmic reticulum of isolated rat skeletal myofibres, were investigated. Aquaporin 4 (AQP4) water channels are arranged as higher order oligomers of several sizes in the sarcolemma and in the T tubules. These oligomers, however, were absent from many specialized micro- and- macrodomains. The distribution of AQP4 coincided with that of a highly organized protein assembly, the dystrophin glycoprotein complex (DGC), in the sarcolemma. A chimaeric venus-AQP4 was equally mobile in the T tubules and sarcolemma, but the anchoring mechanisms of the protein appeared to be different. In contrast to AQP4, the proteins resident in cholesterol and sphingolipid-based microdomains, known as rafts, also occupied DGC deficient areas, which surround the T tubule openings. Indeed, flotillin-1 rafts were located in the neck portions of the T tubules. The rafts defined by the influenza haemagglutinin (HA) also resided in DGC deficient areas, but at the borders of the DGC area. Importantly, of the raft proteins, only the localization of caveolin 3 (CAV3) was dependent on the cholesterol enriched lipid environment, as evidenced by cholesterol depletion experiments and localization studies on a non-raft associated variant of HA. The organization and distribution of membrane associated rough ER (RER) proteins were also analysed. Biochemical detergent extraction analyses and immunofluorescence staining indicated that the ER proteins were assembled as microdomains within the sarcoplasmic reticulum (SR). The microdomains were distributed throughout the SR network and they were capable of protein translocation. Taken together, skeletal myofibres comprise visually distinct microdomains both in the plasma membrane and in the SR. In the plasma membrane, different types of microdomains are not homogenously distributed and function in diverse locations. This may have important physiological implications concerning, among other things, local regulation of ion concentrations and cell signalling cascades. Different constraints ranging from protein-protein interactions to the surrounding lipid environment are important for dictating the observed distribution patterns. / Tiivistelmä Luustolihaksen toimintojen perustana ovat supistumiskykyiset lihassolut, joiden kalvorakenne on järjestynyt erityisellä tavalla ohjaamaan supistusta. Tässä tutkimuksessa analysoitiin proteiini- ja lipidiperustaisten mikroalueiden järjestäytymistä ja tähän vaikuttavia tekijöitä luustolihassolun solukalvolla sekä lihassolun sisäisessä kalvojärjestelmässä, sarkoplasmisessa verkossa (SR). Ensin analysoitiin vesikanavatyyppiä 4 (AQP4), joka oligomerisoituessaan muodostaa erikokoisia mikroalueita. Havaittiin, että AQP4-mikroalueita esiintyy kaikkialla solukalvolla lukuun ottamatta eräitä erilaistuneita mikro- ja makroalueita. AQP4-oligomeerien jakauma solukalvon lateraalisessa osassa, sarkolemmalla, noudatti dystrofiini-glykoproteiinikompleksin jakaumaa. Fluoresoivan venus-AQP4-proteiinin avulla osoitettiin, että proteiinin liikkuvuus oli samanlainen solun sisään ulottuvissa poikkiputkistoissa ja sarkolemmalla, mutta liikkuvuutta rajoittavat tekijät olivat erilaisia näissä solukalvon osissa. Toiseksi analysoitiin kolesteroli- ja sfingolipidipitoisia mikroalueita, kalvolauttoja. Flotilliini-1- ja influenssaviruksen hemagglutiniini (HA) -proteiinia sisältäviä lauttoja esiintyi vain poikkiputkien suuaukkojen alueella, mutta lauttojen jakauma oli erilainen. Lauttojen lipidiympäristöllä ei ollut vaikutusta proteiinien sijaintiin. Tämä osoitettiin kolesterolin poistokokeilla sekä kokeilla, joissa käytettiin mutatoitua HA-proteiinia, joka ei hakeudu kolesteroliympäristöön. Kaveoliini-3-proteiinin sijainti poikkeaa edellä mainituista, ja kolesterolin poisto vaikutti merkittävästi sijainnin määräytymiseen. Kolmanneksi analysoitiin, miten karkean endoplasmakalvoston proteiinit ovat järjestäytyneet SR:ssä. Havaittiin, että endoplasmiset kalvoproteiinit eivät ole homogeenisesti levittäytyneet SR-kalvostoon vaan muodostavat pieniä mikroalueita. Detergenttiuuttoanalyysit osoittivat lisäksi, että näissä mikroalueissa on erilainen lipidikoostumus kuin SR:ssä yleensä. Huomattavaa oli myös, että mikroalueet olivat toiminnallisia kaikkialla SR-kalvostossa. Tulosten perusteella luustolihassolujen kalvojärjestelmä sisältää mikroalueita, joiden jakautuminen vaikuttaa hyvin organisoituneelta. Erityisesti solukalvon mikroalueet esiintyvät tietyillä spesifeillä alueilla, joissa niiden voidaan olettaa toimivan mm. erilaisissa solusignalointitapahtumissa ja paikallisessa ionipitoisuuksien säätelyssä. Eräissä tapauksissa lipidiympäristöllä on merkitystä mikroalueiden sijainnin määräytymisessä, mutta proteiinien sitoutuminen solukalvo- tai solukalvon alaisiin rakenteisiin saattaa myös olla määräävä tekijä.

Musculoskeletal injuries are commonly associated with muscle atrophy as a function of immobilization or change of normal function. For example, injuries to the anterior cruciate ligament (ACL) which may involve ligament reconstruction, results in the "quadriceps avoidance" gait which leads to atrophy of the knee extensormuscles. In these situations it is not clear whether or not the atrophy is associated with loss of specific muscle fibre types with accompanying functional deficits. Such knowledge would be helpful in implementing exercise regimes designed to compensate for loss of particular fibre types. It is believed that isokinetic exercise performed at speeds below 180° per second strengthens type I muscle fibres, and type II fibres at fast speeds. However, there is no evidence to indicate the specific muscle fibre response to different rates of muscle contraction. Identification of muscle fibre type is most directly determined by biopsy technique but is too invasive for a routine measurement. Electromyography median frequency has been used as a non-invasive measure to infer muscle fibre composition in various studies. However, the reliability and accuracy of this technique has been questioned and improvement is necessary. This research was designed to provide a more accurate and reliable protocol for the determination of EMG median frequency which may be used, after validation against more direct biopsy techniques, as a routine method for inferring muscle fibre composition. The investigation also explored the muscular response as measured by EMG median frequency to varying speeds of muscle contraction, fatiguing exercise and atrophy following ACL reconstruction. The ultimate aim of this research was to improve the reliability of the determination of EMG median frequency to enhance its application as a predictor of muscle fibre composition. This provides information which may improve ACL rehabilitation programs designed to restore and prevent specific muscle fibre types loss that have not previously been targeted by current rehabilitation programs. This research was conducted in three studies. Study one determined the stability of the EMG median frequency bilaterally for the quadriceps and hamstrings muscles and identified the mode of contraction associated with the greatest reliability. The strength and EMG median frequency of the vastus lateralis, medial hamstrings and vastus medialis of 55 subjects was determined across 5 speeds from 0° to 240° per second using a Kin-Com isokinetic dynamometer and an EMG data acquisition system. Isometric contraction was found to have the least bilateral discrepancy (4.01% ±3.06) and between trials standard deviation (4.50) in the vastus lateralis, medial hamstrings and vastus medialis. Study two investigated the EMG median frequency changes in the vastus lateralis which occur immediately following different speeds of isokinetic exercise to the point of fatigue in normal subjects. Thirty-four subjects participated in the study, and performed a 90-second period of isokinetic exercise to activate the knee extensors at either 30° or 300° per second. EMG median frequency of the vastus lateralis was determined before, immediately after and 7 minutes after the fatiguing exercise. The percentage drop in EMG median frequency of the vastus medialis was gnificantly (p<0.05) greater after slow speed (27.9%) than fast speed (20.25%) exercise, while no significant difference was found for the percentage drop in extension torque. Full recovery was found 7 minutes after the fatiguing exercise. By reference to previous research showing a relationship between EMG median frequency and muscle fibre type, an increase in activation of type I muscle fibres with slow speed exercise and an increase in type II muscle fibres with fast speed exercise was observed. Study three identified the changes in EMG median frequency following ACL reconstruction and evaluated the bilateral differences in EMG median frequency of the knee muscles. The relationships between EMG median frequency and the measures of knee functional ability, knee muscle strength, age and time since surgery were also investigated. Twelve subjects who had undergone ACL reconstruction using a semitendinosus and gracilis graft 6 to 12 months earlier, participated in the study. EMG median frequency was determined from an 8-second isometric contraction and knee functional ability was assessed using the Cincinnati Rating Scale. Bilateral EMG median frequency shifts were inconsistent among subjects. On the basis of previous research which indicated a relationship between EMG median frequency and fibre type, no consistent pattern of muscular fibre type atrophy subsequent to ACL reconstruction occurred within 6 to 12 months (ranged from -43 to 57 Hz). Additionally, no significant correlations were found between the EMG median frequency and the knee functional score and knee extension and flexion torques, age, time since operation and the bilateral differences in EMG median frequency. The results of this investigation will serve to improve the reliability of EMG median frequency across a range of conditions in which it has been evaluated. Further research is needed to confirm the relationship between EMG median frequency and direct observations of muscle fibre composition to improve the predictive value of this measure. Following this validation it will be possible to evaluate the bilateral EMG median frequency shift to infer the type of muscle fibre atrophy, and use this measure in determining the efficacy of specific rehabilitation programs. In conclusion * An 8-second isometric contraction is recommended for determining EMG median frequency. * EMG median frequency of a muscle decreases significantly more after slow fatiguing exercise than after fast speed fatiguing exercise. * There was no generalised bilateral EMG median frequency shift found in a group of subjects 6 to 12 months following semitendinosus and gracilis graft ACL reconstruction. * The results of this study will serve to improve the reliability of procedures used to determine EMG median frequency under a range of different contractile conditions. The EMG median frequency changes in response to these conditions require further validations with muscle biopsy in future.