Casitas b-lineage lymphoma-deficient mice are pretected against high-fat diet-induuced obesity and insulin resistance

Casitas b-lineage lymphoma (c-Cbl) is a multiadaptor protein with E3-ubiquitin ligase activity involved in regulating the degradation of receptor tyrosine kinases. We have recently reported that c-Cbl^–/– mice exhibit a lean phenotype and enhanced peripheral insulin action likely due to elevated energy expenditure. In the study reported here, we examined the effect of a high-fat diet on energy homeostasis and glucose metabolism in these animals. When c-Cbl^–/– mice were fed a high-fat diet for 4 weeks, they maintained hyperphagia, higher whole-body oxygen consumption (27%), and greater activity (threefold) compared with wild-type animals fed the same diet. In addition, the activity of several enzymes involved in mitochondrial fat oxidation and the phosphorylation of acetyl CoA carboxylase was significantly increased in muscle of high-fat–fed c-Cbl–deficient mice, indicating a greater capacity for fat oxidation in these animals. As a result of these differences, fat-fed c-Cbl^–/– mice were 30% leaner than wild-type animals and were protected against high-fat diet–induced insulin resistance. These studies are consistent with a role for c-Cbl in regulating nutrient partitioning in skeletal muscle and emphasize the potential of c-Cbl as a therapeutic target in the treatment of obesity and type 2 diabetes.

c-Cbl-deficient mice have reduced adiposity, higher energy expenditure, and improved peripheral insulin action

Casitas b-lineage lymphoma (c-Cbl) is an E3 ubiquitin ligase that has an important role in regulating the degradation of cell surface receptors. In the present study we have examined the role of c-Cbl in whole-body energy homeostasis. c-Cb^-/- mice exhibited a profound increase in whole-body energy expenditure as determined by increased core temperature and whole-body oxygen consumption. As a consequence, these mice displayed a decrease in adiposity, primarily due to a reduction in cell size despite an increase in food intake. These changes were accompanied by a significant
increase in activity (2- to 3-fold). In addition, cc-Cb-/- mice displayed a marked improvement in whole-body insulin action, primarily due to changes in muscle metabolism. We observed increased protein levels of the insulin receptor (4-fold) and uncoupling protein-3 (2-fold) in skeletal muscle and a significant increase in the phosphorylation of AMP-activated protein kinase and acetyl-CoA carboxylase. These fmdings suggest that c-Cbl plays an integral role in whole-body fuel homeostasis by regulating whole-body energy expenditure and insulin action.

Modelling blackwater: predicting water quality during flooding of lowland river forests

The blackwater model was developed to predict adverse water quality associated with flooding of the Barmah-Millewa Forests on the River Murray. Specifically, the model examines the likelihood and severity of blackwater events—high dissolved organic carbon associated with low dissolved oxygen. The Barmah-Millewa Forests are dominated by an overstorey of River Red Gum (Eucalyptus camaldulensis) and the litter from these trees contributes a substantial proportion of the pulse of dissolved organic matter released from the floodplain during flooding. This model examines rates of litter accumulation and decay on the floodplain (prior to and during flooding), rates of carbon leaching, microbial degradation, oxygen consumption, reaeration processes and the effects of flow on the concentrations of dissolved organic carbon and dissolved oxygen in the water column (both on the floodplain and in the river channel downstream). The model has been calibrated with data from two blackwater events that have taken place in these forests within the last 5 years. Scenario testing with the model highlights the particularly important roles of flow and temperature in the development of anoxia. Pooled floods and those in the warmest months of the year are substantially more likely to result in blackwater events than floods in cooler times of the year and involving more water exchange between the river channel and the floodplain.

Exercise increases serum Hsp72 in humans

Recent evidence suggests that heat shock proteins (Hsps) may have an important systemic role as a signal to activate the immune system. Since acute exercise is known to induce Hsp72 (the inducible form of the 70-kDa family of Hsp) in a variety of tissues including contracting skeletal muscle, we hypothesized that such exercise would result in the release of Hsp72 from stressed cells into the blood. Six humans (5 males, 1 female) ran on a treadmill for 60 minutes at a workload corresponding to 70% of their peak oxygen consumption. Blood was sampled from a forearm vein at rest (R), 30 minutes during exercise, immediately postexercise (60 minutes), and 2, 8, and 24 hours after exercise. These samples were analyzed for serum Hsp72 protein. In addition, plasma creatine kinase (CK) was measured at these time points as a crude marker of muscle damage. With the exception of the sample collected at 30 minutes, muscle biopsies (n = 5 males) were also obtained from the vastus lateralis at the time of blood sampling and analyzed for Hsp72 gene and protein expression. Serum Hsp72 protein increased from rest, both during and after exercise (0.13 0.10 vs 0.87 ± 0.24 and 1.02 ± 0.41 ng/mL at rest, 30 and 60 minutes, respectively, P < 0.05, mean SE). In addition, plasma CK was elevated (P < 0.05) 8 hours postexercise. Skeletal muscle Hsp72 mRNA expression increased 6.5-fold (P < 0.05) from rest 2 hours postexercise, and although there was a tendency for Hsp72 protein expression to be elevated 2 and 8 hours following exercise compared with rest, results were not statistically significant. The increase in serum Hsp72 preceded any increase in Hsp72 gene or protein expression in contracting muscle, suggesting that Hsp72 was released from other tissues or organs. This study is the first to demonstrate that acute exercise can increase Hsp72 in the peripheral circulation, suggesting that during stress these proteins may indeed have a systemic role.

End-Permian catastrophe by a bolide impact: evidence of a gigantic release of sulfur from the mantle

Our studies in southern China have revealed a remarkable sulfur and strontium isotope excursion at the end of the Permian, along with a coincident concentration of impact- metamorphosed grains and kaolinite and a significant decrease in manganese, phosphorous, calcium, and microfossils (foraminifera). These data suggest that an asteroid or a comet hit the ocean at the end of Permian time and caused a rapid and massive release of sulfur from the mantle to the ocean-atmosphere system, leading to significant oxygen consumption, acid rain, and the most severe biotic crisis in the history of life on Earth.

Live high: train low increases muscle buffer capacity and submaximal cycling efficiency

This study investigated whether hypoxic exposure increased muscle buffer capacity (βm) and mechanical efficiency during exercise in male athletes. A control (CON, n=7) and a live high:train low group (LHTL, n=6) trained at near sea level (600 m), with the LHTL group sleeping for 23 nights in simulated moderate altitude (3000 m). Whole body oxygen consumption (V˙O₂) was measured under normoxia before, during and after 23 nights of sleeping in hypoxia, during cycle ergometry comprising 4×4-min submaximal stages, 2-min at 5.6 ± 0.4 W kg^–1, and 2-min 'all-out' to determine total work and V˙O_2peak. A vastus lateralis muscle biopsy was taken at rest and after a standardized 2-min 5.6 ± 0.4 W kg^–1 bout, before and after LHTL, and analysed for βm and metabolites. After LHTL, βm was increased (18%, P < 0.05). Although work was maintained, V˙O_2peak fell after LHTL (7%, P < 0.05). Submaximal V˙O₂ was reduced (4.4%, P < 0.05) and efficiency improved (0.8%, P < 0.05) after LHTL probably because of a shift in fuel utilization. This is the first study to show that hypoxic exposure, per se, increases muscle buffer capacity. Further, reduced V˙O₂ during normoxic exercise after LHTL suggests that improved exercise efficiency is a fundamental adaptation to LHTL.

Chronic intermittent hypoxia and incremental cycling exercise independently depress muscle in vitro maximal Na+-K+-ATPase activity in well-trained athletes

Athletes commonly attempt to enhance performance by training in normoxia but sleeping in hypoxia [live high and train low (LHTL)]. However, chronic hypoxia reduces muscle Na⁺-K⁺-ATPase content, whereas fatiguing contractions reduce Na⁺-K⁺-ATPase activity, which each may impair performance. We examined whether LHTL and intense exercise would decrease muscle Na⁺-K⁺-ATPase activity and whether these effects would be additive and sufficient to impair performance or plasma K⁺ regulation. Thirteen subjects were randomly assigned to two fitness-matched groups, LHTL (n = 6) or control (Con, n = 7). LHTL slept at simulated moderate altitude (3,000 m, inspired O₂ fraction = 15.48%) for 23 nights and lived and trained by day under normoxic conditions in Canberra (altitude ~600 m). Con lived, trained, and slept in normoxia. A standardized incremental exercise test was conducted before and after LHTL. A vastus lateralis muscle biopsy was taken at rest and after exercise, before and after LHTL or Con, and analyzed for maximal Na⁺-K⁺-ATPase activity [K⁺-stimulated 3-O-methylfluorescein phosphatase (3-O-MFPase)] and Na⁺-K⁺-ATPase content ([³H]ouabain binding sites). 3-O-MFPase activity was decreased by –2.9 ± 2.6% in LHTL (P < 0.05) and was depressed immediately after exercise (P < 0.05) similarly in Con and LHTL (–13.0 ± 3.2 and –11.8 ± 1.5%, respectively). Plasma K⁺ concentration during exercise was unchanged by LHTL; [3H]ouabain binding was unchanged with LHTL or exercise. Peak oxygen consumption was reduced in LHTL (P < 0.05) but not in Con, whereas exercise work was unchanged in either group. Thus LHTL had a minor effect on, and incremental exercise reduced, Na⁺-K⁺-ATPase activity. However, the small LHTL-induced depression of 3-O-MFPase activity was insufficient to adversely affect either K⁺ regulation or total work performed.

Fenitrothion, an organophosphate, affects running endurance but not aerobic capacity in fat-tailed dunnarts (Sminthopsis crassicaudata)

We measured aerobic metabolism during cold exposure and exercise performance (run duration and oxygen consumption while running at 1 m s⁻¹) in the fat-tailed dunnart Sminthopsis crassicaudata, a dasyurid marsupial, before and after ingestion of 30 mg kg⁻¹ of fenitrothion, an organophosphate (OP) pesticide. Running endurance of OP-exposed animals was less than half that of control animals over the first 3 days after dosing and 55% of control animal endurance on day 5 post-dose. Despite these declines, peak metabolic rate at this running speed (9.3 times basal metabolic rate; BMR) was unaffected by OP exposure. Peak metabolic rate (PMR) and cumulative oxygen consumption during a 1-h exposure to conditions equivalent to −20 °C did not differ between OP-treated and control dunnarts, with PMR averaging 11 times BMR. We conclude that fenitrothion-induced exercise fatigue is not due to limitations in oxygen or substrate delivery to muscle or in their uptake per se, but more likely relates to decreased ability to sustain high-frequency neuromuscular function. The persistence of locomotor impairment following OP exposure in otherwise asymptomatic animals emphasizes the importance of using performance-based measures when characterising sublethal effects of pesticide exposure in an ecological context.

Testosterone effects on avian basal metabolic rate and aerobic performance : facts and artefacts

We examined the effects of cage size and testosterone (T) levels on basal and peak metabolic rates (BMR and PMR, respectively) and on pectoral and leg muscle masses of male house sparrows (Passer domesticus). Birds were housed either in small birdcages or in flight aviaries for at least 2 weeks prior to the initial metabolic evaluations. They were then implanted with either empty or T-filled silastic capsules and remeasured 5–6 weeks later. Birds treated with single T implants achieved breeding levels (4–6 ng/mL) and one group given double implants reached 10 ng/mL. There was no effect of T on BMR or PMR in any group studied, but there was an effect of caging. Caged birds showed significant reductions in PMR over the course of captivity, whereas PMR in aviary-housed birds were indistinguishable from their free-living counterparts. Testosterone treatment significantly increased leg muscle mass in caged birds, but had no effect on muscle mass in aviary-housed sparrows. We conclude that testosterone has no direct effect on sparrow metabolic rate or muscle mass, but may interact with cage conditions to produce indirect changes to these variables.

Effect of posture on high-intensity constant-load cycling performance in men and women

The time sustained during a graded cycle exercise is ~10% longer in an upright compared with a supine posture. However, during constant-load cycling this effect is unknown. Therefore, we tested the postural effect on the performance of high-intensity constant-load cycling. Twenty-two active subjects (11 men, 11 women) performed two graded tests (one upright, one supine), and of those 22, 10 subjects (5 men, 5 women) performed three high-intensity constant-load tests (one upright, two supine). To test the postural effect on performance at the same absolute intensity, during the upright and one of the supine constant-load tests subjects cycled at 80% of the peak power output achieved during the upright graded test. To test the postural effect on performance at the same relative intensities, during the second supine test subjects cycled at 80% of the peak power output achieved during the supine graded test. Exercise time on the graded and absolute intensity constant-load tests for all subjects was greater (P<0.05) in the upright compared with supine posture (17.9±3.5 vs. 16.1±3.1 min for graded; 13.2±8.7 vs. 5.2±1.9 min for constant-load). This postural effect at the same absolute intensity was larger in men (19.4±8.5 upright vs. 6.6±1.6 supine, P<0.001) than women (7.1±2 upright vs. 3.9±1.4 supine, P>0.05) and it was correlated (P<0.05) with both the difference in VO₂ between positions during the first minute of exercise (r=0.67) and the height of the subjects (r=0.72). In conclusion, there is a very large postural effect on performance during constant-load cycling exercise and this effect is significantly larger in men than women.

Energetics of interlimb coordination

While the traditional dependent variables of motor skill learning are accuracy and consistency of movement outcome, there has been increasing interest in aspects of motor performance that are described as reflecting the ‘energetics’ of motor behaviour. One defining characteristic of skilled motor performance is the ability to complete the task with minimum energy expenditure (Sparrow & Newell, 1998). A further consideration is that movements also have costs in terms of cognitive ‘effort’ or ‘energy’. The present project extends previous work on energy expenditure and motor skill learning within a coordination dynamics framework. From the dynamic pattern perspective, a coordination pattern lowest on the 11KB model potential curve (Haken, Kelso & Bunz, 1985) is more stable and least energy is required to maintain pattern stability (Temprado, Zanone, Monno & Laurent, 1999). Two experiments investigated the learning of stable and unstable coordination patterns with high metabolic energy demand. An experimental task was devised by positioning two cycle ergometers side-by-side, placing one foot on each, with the pedals free to move independently at any metronome-paced relative phase, Experiment 1 investigated practice-related changes to oxygen consumption, heart rate, relative phase, reaction time and muscle activation (EMG) as participants practiced anti-phase, in-phase and 90°-phase cycling. Across six practice trials metabolic energy cost reduced and AE and VE of relative phase declined. The trend in the metabolic and reaction time data and percent co-contraction of muscles was for the in-phase cycling to demonstrate the highest values, anti-phase the lowest and 90°-phase cycling in-between. It was found that anti- and in-phase cycling were both kinematically stable but anti-phase coordination revealed significantly lower metabolic energy cost. It was, therefore, postulated that of two equally stable coordination patterns, that associated with lower metabolic energy expenditure would constitute a stronger attractor. Experiment 2 was designed to determine whether a lower or higher energy-demanding coordination pattern was a stronger attractor by scanning the attractor layout at thirty-degree intervals from 0° to 330°. The initial attractor layout revealed that in-phase was most stable and accurate, but the remaining coordination patterns were attracted to the low energy cost anti-phase cycling. In Experiment 2 only 90°- phase cycling was practiced with a post-test attractor layout scan revealing that 90°-phase and its symmetrical partner 270°-phase had become attractors of other coordination patterns. Consistent with Experiment 1, practicing 90°-phase cycling revealed a decline in AE and VE and a reduction in metabolic and cognitive cost. Practicing 90°-phase cycling did not, however, destabilise the in-phase or anti-phase coordination patterns either kinematically or energetically. In summary, the findings suggest that metabolic and mental energy can be considered different representations of a ‘global’ energy expenditure or ‘energetic’ phenomenon underlying human coordination. The hypothesis that preferred coordination patterns emerge as stable, low-energy solutions to the problem of inter-and intra-limb coordination is supported here in showing that the low-energy minimum of coordination dynamics is also an energetic minimum.

Impaired K+ regulation contributes to exercise limitation in end-stage renal failure

Background

Patients with end-stage renal failure (ESRF) exhibit grossly impaired maximal exercise performance. This study investigated whether K+ regulation during exercise is impaired in ESRF and whether this is related to reduced exercise performance.

Methods

Nine stable hemodialysis patients and eight controls (CON) performed incremental cycling exercise to volitional fatigue, with measurement of peak oxygen consumption (VdotO2 peak). Arterial blood was sampled during and following exercise and analyzed for plasma [K+] (PK).

Results

The VdotO2 peak was approximately 44% less in ESRF than in CON (P < 0.001), whereas peak exercise PK was greater (7.23 plusminus 0.38 vs. 6.23 plusminus 0.14 mmol dot L-1, respectively, P < 0.001). In ESRF, the rate of rise in PK during exercise was twofold greater (0.43 plusminus 0.05 vs. 0.23 plusminus 0.03 mmol dot L-1dotmin-1, P < 0.005) and the ratio of rise in PK relative to work performed was 3.7-fold higher (90.1 plusminus 13.5 vs. 24.7 plusminus 3.3 nmol dot L-1dot J-1, P < 0.001). A strong inverse relationship was found between VdotO2 peak and the DeltaPKdot work-1 ratio (r = -0.80, N = 17, P < 0.001).

Conclusions

Patients with ESRF exhibit grossly impaired extrarenal K+ regulation during exercise, demonstrated by an excessive rise in PK relative to work performed. We further show that K+ regulation during exercise was correlated with aerobic exercise performance. These results suggest that disturbed K+ regulation in ESRF contributes to early muscle fatigue during exercise, thus causing reduced exercise performance.

Impaired muscle Ca2+ and K+ regulation contribute to poor exercise performance post-lung transplantation

Lung transplant recipients (LTx) exhibit marked peripheral limitations to exercise. We investigated whether skeletal muscle Ca2+ and K+ regulation might be abnormal in eight LTx and eight healthy controls. Peak oxygen consumption and arterialized venous plasma [K+] (where brackets denote concentration) were measured during incremental exercise. Vastus lateralis muscle was biopsied at rest and analyzed for sarcoplasmic reticulum Ca2+ release, Ca2+ uptake, and Ca2+-ATPase activity rates; fiber composition; Na+-K+-ATPase (K+-stimulated 3-O-methylfluorescein phosphatase) activity and content ([3H]ouabain binding sites); as well as for [H+] and H+-buffering capacity. Peak oxygen consumption was 47% less in LTx (P < 0.05). LTx had lower Ca2+ release (34%), Ca2+ uptake (31%), and Ca2+-ATPase activity (25%) than controls (P < 0.05), despite their higher type II fiber proportion (LTx, 75.0 ± 5.8%; controls, 43.5 ± 2.1%). Muscle [H+] was elevated in LTx (P < 0.01), but buffering capacity was similar to controls. Muscle 3-O-methylfluorescein phosphatase activity was 31% higher in LTx (P < 0.05), but [3H]ouabain binding content did not differ significantly. However, during exercise, the rise in plasma [K+]-to-work ratio was 2.6-fold greater in LTx (P < 0.05), indicating impaired K+ regulation. Thus grossly subnormal muscle calcium regulation, with impaired potassium regulation, may contribute to poor muscular performance in LTx.