Aspects of the nutritional physiology of the perlemoen Haliotis midae (L.) and red abalone H. rufescens (Swainson)

The source of abalone for human consumption has shown a dramatic shift away from wild-capture fisheries in the last 30 years, with over 90% of global production now coming from aquaculture. Farmers initially relied on the natural food of abalone (macroalgae) as a culture feed, though in regions where macroalgae availability was limiting, the need to develop formulated feeds was evident. Extensive research effort has led to the development of a number of formulated feed products currently employed in the industry. These feeds, however, differ markedly from the mixed macroalgal diets that abalone have evolved to utilise, particularly in terms of protein content and carbohydrate structure. The degree to which the nutritional physiology of abalone responds to these novel formulated diets, with and without macroalgal supplementation, was investigated in the current study. A multifaceted approach, combining growth trials, stable isotope nutrient tracers and metabolic experiments, was employed to gain insight into the post-absorption dynamics and utilisation of dietary nutrients under varying dietary regimes of fresh macroalgae and formulated feed. Growth trials conducted with both Haliotis rufescens and H. midae showed significantly higher growth and protein utilisation efficiency for abalone fed macroalgal diets compared to formulated feeds. Furthermore, when formulated feeds were supplemented with macroalgae to form combination diets, growth and the utilization of protein was improved compared to the formulated-feed-only diet. The poor utilisation of protein by H. midae fed the formulated feed could be traced, using a method combining stable isotope bio-markers with a Bayesian mixing model (SIAR), to the low incorporation of the fishmeal component of protein in the diet. The marked postprandial drop in the O:N ratio on abalone fed formulated feeds indicate that the protein was being diverted into catabolic metabolic pathways. The metabolic cost of digestion, termed specific dynamic action (SDA), was negated as a factor in the improved growth of abalone fed macroalgal diets, with the SDA coefficient 2.1 times that observed for formulated feed. Furthermore, the postprandial haemolymph glucose concentration (HGC) in H. midae was elevated when fed formulated feed compared to macroalgae. The high levels of circulating glucose are likely a result of the structure the carbohydrate source in formulated feeds and stimulate the deposition of glycogen through the allosteric control of glycogen synthase. Formulated feeds produced higher cooked meat yields in canning simulation trials, suggesting that muscle glycogen content may indirectly play a role in increasing canning yields through the displacement of collagen. The results of these empirical studies are synthesised under key themes, discussed within the context of their potential commercial relevance and future research directions are highlighted

Classifying training drills based on movement demands in Australian Football

Time-motion data was used to classify a selection of training drills. Ten midfielders (age=23.8±1.8yr; height=183.9±3.8cm; mass=83.2±5.0 kg) from an Australian Football League team participated in 17 training drills and four quarters of an official competitive match. Heart rate and time-motion data were collected using Global Positioning and Heart Rate Systems. Cluster analysis of mean distance travelled in the seven velocity zones identified three clusters: 1) game-specific conditioning; 2) skill refining/moderate intensity dominant; and 3) skill refining/low intensity dominant. Differences between the three clusters in distance travelled at the speed zones were confirmed using one-way ANOVA. Differences between clusters were also assessed for number of efforts in velocity zones and percentage time in heart rate zones. When compared to drills with a focus on skill refining or performed on a reduced playing area, drills utilising the entire playing field better replicated the movement characteristics of competitive game play

Skeletal muscle AMPK is essential for the maintenance of FNDC5 expression

Fibronectin type III domain‐containing protein 5 (FNDC5) expression is controlled by the transcriptional co‐activator, peroxisome proliferator‐activated receptor gamma, coactivator 1 alpha (PGC1α). FNDC5 expression has been shown to be increased in muscle in response to endurance exercise in some but not all studies, therefore a greater understanding of the mechanisms controlling this process are needed. The AMP‐activated protein kinase (AMPK) is activated by exercise in an intensity dependent manner and is an important regulator of PGC1α activity; therefore, we explored the role of AMPK in the regulation of FNDC5 using AMPK β1β2 double muscle‐null mice (AMPK DMKO), which lack skeletal muscle AMPK activity. We found that FNDC5 expression is dramatically reduced in resting muscles of AMPK DMKO mice compared to wild‐type littermates. In wild‐type mice, activating phosphorylation of AMPK was elevated immediately post contraction and was abolished in muscle from AMPK DMKO mice. In contrast, PGC1α was increased in both wild‐type and AMPK DMKO mice 3 h post contraction but FNDC5 protein expression was not altered. Lastly, acute or chronic activation of AMPK with the pharmacological AMPK activator AICAR did not increase PGC1α or FNDC5 expression in muscle. These data indicate that skeletal muscle AMPK is required for the maintenance of basal FNDC5 expression

Versatility of Fear-Potentiated Startle Paradigms for Assessing Human Conditioned Fear Extinction and Return of Fear

Fear conditioning methodologies have often been employed as testable models for assessing learned fear responses in individuals with anxiety disorders such as post-traumatic stress disorder (PTSD) and specific phobia. One frequently used paradigm is measurement of the acoustic startle reflex under conditions that mimic anxiogenic and fear-related conditions. For example, fear-potentiated startle is the relative increase in the frequency or magnitude of the acoustic startle reflex in the presence of a previously neutral cue (e.g., colored shape; termed the conditioned stimulus or CS+) that has been repeatedly paired with an aversive unconditioned stimulus (e.g., airblast to the larynx). Our group has recently used fear-potentiated startle paradigms to demonstrate impaired fear extinction in civilian and combat populations with PTSD. In the current study, we examined the use of either auditory or visual CSs in a fear extinction protocol that we have validated and applied to human clinical conditions. This represents an important translational bridge in that numerous animal studies of fear extinction, upon which much of the human work is based, have employed the use of auditory CSs as opposed to visual CSs. Participants in both the auditory and visual groups displayed robust fear-potentiated startle to the CS+, clear discrimination between the reinforced CS+ and non-reinforced CS−, significant extinction to the previously reinforced CS+, and marked spontaneous recovery. We discuss the current results as they relate to future investigations of PTSD-related impairments in fear processing in populations with diverse medical and psychiatric histories

Electrochemical titrations and reaction time courses monitored in situ by magnetic circular dichroism spectroscopy

Magnetic circular dichroism (MCD) spectra, at ultraviolet–visible or near-infrared wavelengths (185–2000 nm), contain the same transitions observed in conventional absorbance spectroscopy, but their bisignate nature and more stringent selection rules provide greatly enhanced resolution. Thus, they have proved to be invaluable in the study of many transition metal-containing proteins. For mainly technical reasons, MCD has been limited almost exclusively to the measurement of static samples. But the ability to employ the resolving power of MCD to follow changes at transition metal sites would be a potentially significant advance. We describe here the development of a cuvette holder that allows reagent injection and sample mixing within the 50-mm-diameter ambient temperature bore of an energized superconducting solenoid. This has allowed us, for the first time, to monitor time-resolved MCD resulting from in situ chemical manipulation of a metalloprotein sample. Furthermore, we report the parallel development of an electrochemical cell using a three-electrode configuration with physically separated working and counter electrodes, allowing true potentiometric titration to be performed within the bore of the MCD solenoid

The glycolytic enzyme phosphofructokinase-1 assembles into filaments.

Despite abundant knowledge of the regulation and biochemistry of glycolytic enzymes, we have limited understanding on how they are spatially organized in the cell. Emerging evidence indicates that nonglycolytic metabolic enzymes regulating diverse pathways can assemble into polymers. We now show tetramer- and substrate-dependent filament assembly by phosphofructokinase-1 (PFK1), which is considered the "gatekeeper" of glycolysis because it catalyzes the step committing glucose to breakdown. Recombinant liver PFK1 (PFKL) isoform, but not platelet PFK1 (PFKP) or muscle PFK1 (PFKM) isoforms, assembles into filaments. Negative-stain electron micrographs reveal that filaments are apolar and made of stacked tetramers oriented with exposed catalytic sites positioned along the edge of the polymer. Electron micrographs and biochemical data with a PFKL/PFKP chimera indicate that the PFKL regulatory domain mediates filament assembly. Quantified live-cell imaging shows dynamic properties of localized PFKL puncta that are enriched at the plasma membrane. These findings reveal a new behavior of a key glycolytic enzyme with insights on spatial organization and isoform-specific glucose metabolism in cells