Local muscle cooling does not impact expression of mitochondrial-related genes

Recovery that takes place in a cold environment after endurance exercise elevates PGC-1α mRNA whereas ERRα and NRF2 mRNA expression are inhibited. However, the effect of local skeletal muscle cooling on mitochondrial-related gene expression is unknown.PurposeTo determine the impact of local skeletal muscle cooling during recovery from an acute bout of exercise on mitochondrial-related gene expression.MethodsRecreationally-trained male cyclists (n=8, age 25±3 y, height 181±6 cm, weight 79±8 kg, 12.8±3.6% body fat, VO_2peak 4.52±0.88 L·min⁻¹ protocol) completed a 90-min variable intensity cycling protocol followed by 4 h of recovery. During recovery, ice was applied intermittently to one leg (ICE) while the other leg served as a control (CON). Intramuscular temperature was recorded continuously. Muscle biopsies were taken from each vastus lateralis at 4 h post-exercise for the analysis of mitochondrial-related gene expression.ResultsIntramuscular temperature was colder in ICE (26.7±1.1 °C) than CON (35.5±0.1 °C) throughout the 4 h recovery period (p<0.001). There were no differences in expression of PGC-1α, TFAM, NRF1, NRF2, or ERRα mRNA between ICE and CON after the 4 h recovery period.ConclusionLocal muscle cooling after exercise does not impact the expression of mitochondrial biogenesis-related genes compared to recovery from exercise in control conditions. When these data are considered with previous research, the stimuli for cold-induced gene expression alterations may be related to factors other than local muscle temperature. Additionally, different intramuscular temperatures should be examined to determine dose-response of mitochondrial-related gene expression.     

Effects of rapid temperature fluctuations prior to breeding on reproductive efficiency in replacement gilts

Rapidly cooling pigs after heat stress (HS) results in a pathophysiological condition, and because rapid temperature fluctuations may be associated with reduced reproductive success in sows, it lends itself to the hypothesis that these conditions may be linked. Objectives were to determine the effects of rapid cooling on thermal response and future reproductive success in pigs. Thirty-six replacement gilts (137.8±0.9 kg BW) were estrus synchronized and then 14.1±0.4 d after estrus confirmation, pigs were exposed to thermoneutral conditions (TN; n=12; 19.7±0.9°C) for 6 h, or HS (36.3±0.5°C) for 3 h, followed by 3 h of rapid cooling (HSRC; n=12; immediate TN exposure and water dousing) or gradual cooling (HSGC; n=12; gradual decrease to TN conditions) repeated over 2 d. Vaginal (T_V) and gastrointestinal tract temperatures (T_GI) were obtained every 15 min, and blood was collected on d 1 and d 2 during the HS and recovery periods at 180 and 60 min, respectively. Pigs were bred 8.3±0.8 d after thermal treatments over 2 d. Reproductive tracts were collected and total fetus number and viability were recorded 28.0±0.8 d after insemination. HS increased T_V and T_GI (P=0.01; 0.98 °C) in HSRC and HSGC compared to TN pigs. During recovery, T_V was reduced from 15 to 105 min (P=0.01; 0.33 °C) in HSRC compared to HSGC pigs, but no overall differences in T_GI were detected (P<0.05; 39.67 °C). Rapid cooling increased (P<0.05) TNFα compared to HSGC and TN pigs during recovery-d 1 (55.2%), HS-d 2 (35.1%), and recovery-d 2 (64.9%). Viable fetuses tended to be reduced (P=0.08; 10.5%) and moribund fetuses tended to be increased (P=0.09; 159.3%) in HSRC compared to HSGC and TN pigs. In summary, rapid cooling prior to breeding may contribute to reduced fetal viability and reproductive success in pigs.     

Repeatability of a running heat tolerance test

At present there is no standardised heat tolerance test (HTT) procedure adopting a running mode of exercise. Current HTTs may misdiagnose a runner's susceptibility to a hyperthermic state due to differences in exercise intensity. The current study aimed to establish the repeatability of a practical running test to evaluate individual's ability to tolerate exercise heat stress. Sixteen (8M, 8F) participants performed the running HTT (RHTT) (30 min, 9 km h⁻¹, 2% elevation) on two separate occasions in a hot environment (40 °C and 40% relative humidity). There were no differences in peak rectal temperature (RHTT1: 38.82±0.47 °C, RHTT2: 38.86±0.49 °C, Intra-class correlation coefficient (ICC)=0.93, typical error of measure (TEM)=0.13 °C), peak skin temperature (RHTT1: 38.12±0.45, RHTT2: 38.11±0.45 °C, ICC=0.79, TEM=0.30 °C), peak heart rate (RHTT1: 182±15 beats min⁻¹, RHTT2: 183±15 beats min⁻¹, ICC=0.99, TEM=2 beats min⁻¹), nor sweat rate (1721±675 g h⁻¹, 1716±745 g h⁻¹, ICC=0.95, TEM=162 g h⁻¹) between RHTT1 and RHTT2 (p>0.05). Results demonstrate good agreement, strong correlations and small differences between repeated trials, and the TEM values suggest low within-participant variability. The RHTT was effective in differentiating between individuals physiological responses; supporting a heat tolerance continuum. The findings suggest the RHTT is a repeatable measure of physiological strain in the heat and may be used to assess the effectiveness of acute and chronic heat alleviating procedures.     

Effect of the timing of ice slurry ingestion for precooling on endurance exercise capacity in a warm environment

It has been demonstrated that precooling with ice slurry ingestion enhances endurance exercise capacity in the heat. However, no studies have yet evaluated the optimal timing of ice slurry ingestion for precooling. This study aimed to investigate the effects of varying the timing of ice slurry ingestion for precooling on endurance exercise capacity in a warm environment. Ten active male participants completed 3 experimental cycling trials to exhaustion at 55% peak power output (PPO) after 15 min of warm-up at 30% PPO at 30 °C and 80% relative humidity. Three experimental conditions were set: no ice slurry ingestion (CON), pre-warm-up ice slurry ingestion (−1 °C; 7.5 g kg⁻¹) (PRE), and post-warm-up ice slurry ingestion (POST). Rectal and mean skin temperatures at the beginning of exercise in the POST condition (37.1±0.2 °C, 33.8±0.9 °C, respectively) were lower than those in the CON (37.5±0.3 °C; P<0.001, 34.8±0.8 °C; P<0.01, respectively) and PRE (37.4±0.2 °C; P<0.01, 34.6±0.7 °C; P<0.01, respectively) conditions. These reductions increased heat storage capacity and resulted in improved exercise capacity in the POST condition (60.2±8.7 min) compared to that in the CON (52.0±11.9 min; effect size [ES]=0.78) and PRE (56.9±10.4 min; ES=0.34) conditions. Ice slurry ingestion after warm-up effectively reduced both rectal and skin temperatures and increased cycling time to exhaustion in a warm environment. Timing ice slurry ingestion to occur after warm-up may be effective for precooling in a warm environment.     

Thermal energetics of the New-Guinean moss-forest rat (Rattus niobe) in comparison with other tropical murid rodents

The thermal energetics of rodents from cool, wet tropical highlands are poorly known. Metabolic rate, body temperature and thermal conductance were measured in the moss-forest rat, Rattus niobe (Rodentia), a small murid endemic to the highlands of New Guinea. These data were evaluated in the context of the variation observed in the genus Rattus and among tropical murids. In 7 adult R. niobe, basal metabolic rate (BMR) averaged 53.6±6.6 mL O₂ h⁻¹, or 103% of the value predicted for a body mass of 42.3±5.8 g. Compared to other species of Rattus, R. niobe combines a low body temperature (35.5±0.6 °C) and a moderately low minimal wet thermal conductance c_min (5.88±0.7 mL O₂ h⁻¹ °C⁻¹, 95% of predicted) with a small size, all of which lead to reduced energy expenditure in a constantly cool environment. The correlations of mean annual rainfall and temperature, altitude and body mass with BMR, body temperature and c_min were analyzed comparatively among tropical Muridae. Neither BMR, nor c_min or body temperature correlated with ambient temperature or altitude. Some of the factors which promote high BMR in higher latitude habitats, such as seasonal exposure to very low temperature and short reproductive season, are lacking in wet montane tropical forests. BMR increased with rainfall, confirming a pattern observed among other assemblages of mammals. This correlation was due to the low BMR of several desert adapted murids, while R. niobe and other species from wet habitats had a moderate BMR.     

Effects of heat acclimation on sweating during graded exercise until exhaustion

The aim of the present study was to test the hypothesis that the sweating during graded exercise until exhaustion in a temperate environment would be greater after heat acclimation. Six healthy young males performed an exercise–heat stress acclimation protocol during 9 days. Before (PRE) and after (POS) the acclimation protocol they performed a graded exercise until exhaustion and the sweat loss during exercise increased after acclimation (3.94±1.10, PRE, and 4.86±1.70 g m⁻² min⁻¹, POS; p<0.05). The results showed that daily prolonged exposures to exercise-heat stress increased sweating during a graded and short duration exercise in a temperate environment.     

GLP-1(32–36)amide,a novel pentapeptide cleavage product of GLP-1, modulates whole body glucose metabolism in dogs

We have previously demonstrated in human subjects who under euglycemic clamp conditions GLP-1(9–36)amide infusions inhibit endogenous glucose production without substantial insulinotropic effects. An earlier report indicates that GLP-1(9–36)amide is cleaved to a nonapeptide, GLP-1(28–36)amide and a pentapeptide GLP-1(32–36)amide (LVKGR amide). Here we study the effects of the pentapeptide on whole body glucose disposal during hyperglycemic clamp studies. Five dogs underwent indwelling catheterizations. Following recovery, the dogs underwent a 180 min hyperglycemic clamp (basal glucose +98 mg/dl) in a cross-over design. Saline or pentapeptide (30 pmol kg⁻¹ min⁻¹) was infused during the last 120 min after commencement of the hyperglycemic clamp in a primed continuous manner. During the last 30 min of the pentapeptide infusion, glucose utilization (M) significantly increased to 21.4 ± 2.9 mg kg⁻¹ min⁻¹compared to M of 14.3 ± 1.1 mg kg⁻¹ min⁻¹ during the saline infusion (P = 0.026, paired t-test; P = 0.062, Mann–Whitney U test). During this interval, no significant differences in insulin (26.6 ± 3.2 vs. 23.7 ± 2.5 μU/ml, P = NS) or glucagon secretion (34.0 ± 2.1 vs. 31.7 ± 1.8 pg/ml, P = NS) were observed. These findings demonstrate that under hyperglycemic clamp studies the pentapeptide modulates glucose metabolism by a stimulation of whole-body glucose disposal. Further, the findings suggest that the metabolic benefits previously observed during GLP-1(9–36)amide infusions in humans might be due, at least in part, to the metabolic effects of the pentapeptide that is cleaved from the pro-peptide, GLP-1(9–36)amide in the circulation.     

Ergogenic and metabolic effects of oral glucocorticoid intake during repeated bouts of high-intensity exercise

All systemically administered glucocorticoids (GC) are prohibited in-competition, because of the potential ergogenic effects. Although short-term GC intake has been shown to improve performance during submaximal exercise, literature on its impact during brief intense exercise appears to be very scant. The purpose of this study was to examine the ergogenic and metabolic effects of prednisone during repeated bouts of high-intensity exercise. In a double-blind randomized protocol, ten recreational male athletes followed two 1-week treatments (Cor: prednisone, 60 mg/day or Pla: placebo). At the end of each treatment, they hopped on their dominant leg for 30 s three times consecutively and then hopped until exhaustion, with intervals of 5 min of passive recovery. Blood and saliva samples were collected at rest and 3 min after each exercise bout to determine the lactate, interleukin-6, interleukin-10, TNF-alpha, DHEA and testosterone values. The absolute peak force of the dominant leg was significantly increased by Cor but only during the first 30-s hopping bout (p < 0.05), whereas time to exhaustion was not significantly changed after Cor treatment vs Pla (Pla: 119.9 ± 24.7; Cor: 123.1 ± 29.5 s). Cor intake lowered basal and end-exercise plasma interleukin-6 and saliva DHEA (p < 0.01) and increased interleukin-10 (p < 0.01), whereas no significant change was found in blood lactate and TNF-alpha or saliva testosterone between Pla and Cor. According to these data, short-term glucocorticoid intake did not improve endurance performance during repeated bouts of high-intensity exercise, despite the significant initial increase in absolute peak force and anti-inflammatory effect.     

A high pressure liquid chromatography-based assay for glutathione-S-transferase class distinction assay

In order to expedite the process of classification of the members of the family of glutathione-S-transferases (GSTs) high performance liquid chromatography with photodiode array detection (HPLC-PDA) was used as a means for measuring enzymatic activity. The GST chosen for the development of the HPLC-PDA technique was from equine liver (E-GST). The characterizing substrates, ethacrynic acid (EA) and bromosulfophthalein (BSP), along with previously gathered characterization data allowed for the distinction of α, μ or π-class enzymes. In an initial characterization of the previously unclassified E-GST it was determined that the enzyme was of the π-class with specific activities of 0.062, ± 0.0015 μmol min^− 1 mg^− 1 and 0.0019, ± 0.00064 μmol min^− 1 mg^− 1 for EA and BSP, respectively. Finally, the activity of the E-GST with the EA and BSP substrates, was measured by HPLC-PDA, and was found to be 0.027, ± 0.003 μmol min^− 1 mg^− 1 and 0.002, ± 0.0005 μmol min^− 1 mg^− 1, respectively. While the HPLC-PDA data do not mirror the spectrophotometric results quantitatively the overall response by the E-GST was the same. In general, the E-GSTs were shown to belong to the π-class when characterized by HPLC-PDA due to an EA specific activity greater than 0.01 μmol min^− 1 mg^− 1 and a negligible BSP activity (≤ 0.002 μmol min^− 1 mg^− 1).     

Performance limitation and the role of core temperature when wearing light-weight workwear under moderate thermal conditions

The objective of this investigation was to achieve an understanding about the relationship between heat stress and performance limitation when wearing a two-layerfire-resistant light-weight workwear (full-clothed ensemble) compared to an one-layer short sports gear (semi-clothed ensemble) in an exhaustive, stressful situation under moderate thermal condition (25 °C). Ten well trained male subjects performed a strenuous walking protocol with both clothing ensembles until exhaustion occurred in a climatic chamber. Wearing workwear reduced the endurance performance by 10% (p=0.007) and the evaporation by 21% (p=0.003), caused a more pronounced rise in core temperature during submaximal walking (0.7±0.3 vs. 1.2±0.4 °C; p≤0.001) and from start till exhaustion (1.4±0.3 vs. 1.8±0.5 °C; p=0.008), accelerated sweat loss (13±2 vs. 15±3 g min⁻¹; p=0.007), and led to a significant higher heart rate at the end of cool down (103±6 vs. 111±7 bpm; p=0.004). Correlation analysis revealed that core temperature development during submaximal walking and evaporation may play important roles for endurance performance. However, a critical core temperature of 40 °C, which is stated to be a crucial factor for central fatigue and performance limitation, was not reached either with the semi-clothed or the full-clothed ensemble (38.3±0.4 vs. 38.4±0.5 °C). Additionally, perceived exertion did not increase to a higher extent parallel with the rising core temperature with workwear which would substantiate the critical core temperature theory. In conclusion, increased heat stress led to cardiovascular exercise limitation rather than central fatigue.     

Mathematical modeling of Microcystis aeruginosa growth and [D-Leu1] microcystin-LR production in culture media at different temperatures

The effect of temperature (26 °C, 28 °C, 30 °C and 35 °C) on the growth of native CAAT-3-2005 Microcystis aeruginosa and the production of Chlorophyll-a (Chl-a) and Microcystin-LR (MC-LR) were examined through laboratory studies. Kinetic parameters such as specific growth rate (μ), lag phase duration (LPD) and maximum population density (MPD) were determined by fitting the modified Gompertz equation to the M. aeruginosa strain cell count (cells mL⁻¹). A 4.8-fold increase in μ values and a 10.8-fold decrease in the LPD values were found for M. aeruginosa growth when the temperature changed from 15 °C to 35 °C. The activation energy of the specific growth rate (Eμ) and of the adaptation rate (E₁/LPD) were significantly correlated (R² = 0.86). The cardinal temperatures estimated by the modified Ratkowsky model were minimum temperature = 8.58 ± 2.34 °C, maximum temperature = 45.04 ± 1.35 °C and optimum temperature = 33.39 ± 0.55 °C.Maximum MC-LR production decreased 9.5-fold when the temperature was increased from 26 °C to 35 °C. The maximum production values were obtained at 26° C and the maximum depletion rate of intracellular MC-LR was observed at 30–35 °C. The MC-LR cell quota was higher at 26 and 28 °C (83 and 80 fg cell⁻¹, respectively) and the MC-LR Chl-a quota was similar at all the different temperatures (0.5–1.5 fg ng⁻¹).The Gompertz equation and dynamic model were found to be the most appropriate approaches to calculate M. aeruginosa growth and production of MC-LR, respectively. Given that toxin production decreased with increasing temperatures but growth increased, this study demonstrates that growth and toxin production processes are uncoupled in M. aeruginosa. These data and models may be useful to predict M. aeruginosa bloom formation in the environment.     

Thermal tolerance and preferred temperature range of juvenile meagre acclimated to four temperatures

The present study reports the temperature tolerance, estimated using dynamic and static methodologies, and preferred temperature range, based on oxygen consumption rate (OCR), of juvenile meagre (Argyrosomus regius) (Asso, 1801) (3.4±0.9 g) after 30 days of acclimation at 18, 22, 26 and 30 °C. Meagre has dynamic and static thermal tolerance zones of 551 °C² and 460 °C², respectively and is a low resistance fish species, with a resistance zone area of 87 °C². The OCR of juvenile meagre at the above acclimation temperatures was 370, 410, 618 and 642 mg h⁻¹ kg⁻¹, respectively, and is significantly different (P<0.0001, n=20). The fact that OCR increases by rising temperatures and gradually decreases after 26 °C indicates that the preferred temperature range of juvenile meagre is between 26 and 30 °C. Our study suggests that meagre is unable to respond to low and high temperature variation in aquaculture facilities or its natural habitats.     

A reduced core to skin temperature gradient,not a critical core temperature,affects aerobic capacity in the heat

The purpose of this study was to determine the impact of the core to skin temperature gradient during incremental running to volitional fatigue across varying environmental conditions. A secondary aim was to determine if a “critical” core temperature would dictate volitional fatigue during running in the heat. 60 participants (n=49 male, n=11 female; 24±5 yrs, 177±11 cm, 75±13 kg) completed the study. Participants were uniformly stratified into a specific exercise temperature group (18 °C, 26 °C, 34 °C, or 42 °C) based on a 3-mile run performance. Participants were equipped with core and chest skin temperature sensors and a heart rate monitor, entered an environmental chamber (18 °C, 26 °C, 34 °C, or 42 °C), and rested in the seated position for 10 min before performing a walk/run to volitional exhaustion. Initial treadmill speed was 3.2 km h⁻¹ with a 0% grade. Every 3 min, starting with speed, speed and grade increased in an alternating pattern (speed increased by 0.805 km h⁻¹, grade increased by 0.5%). Time to volitional fatigue was longer for the 18 °C and 26 °C group compared to the 42 °C group, (58.1±9.3 and 62.6±6.5 min vs. 51.3±8.3 min, respectively, p<0.05). At the half-way point and finish, the core to skin gradient for the 18 °C and 26 °C groups was larger compared to 42 °C group (halfway: 2.6±0.7 and 2.0±0.6 vs. 1.3±0.5 for the 18 °C, 26 °C and 42 °C groups, respectively; finish: 3.3±0.7 and 3.5±1.1 vs. 2.1±0.9 for the 26 °C, 34 °C, and 42 °C groups, respectively, p<0.05). Sweat rate was lower in the 18 °C group compared to the 26 °C, 34 °C, and 42 °C groups, 3.6±1.3 vs. 7.2±3.0, 7.1±2.0, and 7.6±1.7 g m⁻² min⁻¹, respectively, p<0.05. There were no group differences in core temperature and heart rate response during the exercise trials. The current data demonstrate a 13% and 22% longer run time to exhaustion for the 18 °C and 26 °C group, respectively, compared to the 42 °C group despite no differences in beginning and ending core temperatures or baseline 3-mile run time. This capacity difference appears to result from a magnified core to skin gradient via an environmental temperature advantageous to convective heat loss, and in part from an increased sweat rate.     

Évaluation de la faisabilité de la tomoscintigraphie myocardique précoce au sestamibi synchronisée à l’électrocardiogramme

BackgroundOur aim was to assess the feasibility of early acquisition (10 min) gated single photon emission computed tomography (SPECT) by comparison to conventional imaging at one hour.Methods and resultsOne hundred and four patients referred for exercise test and SPECT were included. Sequential imaging was started 10 min (SPECT 10) and 60 min (SPECT 60) after injection of the radiotracer (Tc-99m sestamibi). Stress myocardial perfusion was visually analyzed from 10 to 60 min stress by two experienced nuclear-cardiologists. Six patients were further excluded, because of high digestive accumulation: one patient at 10 min, three at 10 and 60 min, two at rest. The participants were classified as follows: group G1 (normal SPECT 10 and 60, n = 53), group G2 (abnormal SPECT 10 and/or SPECT 60, n = 45). The left ventricle ejection fraction (EF) and volumes (end-systolic and end-diastolic volumes, ESV, EDV) were calculated with the Cedars-Sinai program.ResultsQuality imaging was the same at SPECT 10 min and SPECT 60 min. Perfusion scores: G1 10 min = 0,4 versus G1 60 min = 0,4 (p = NS); G2 10 min = 10,2 versus G2 60 min = 10,1 (p = NS); EFG1 10 min = 71 ± 11% versus EFG1 60 min = 68 ± 10% (p = 4E-04); EFG2 10 min = 56 ± 15% versus EFG2 60 min = 53 ± 15% (p = 0,002); EDV G1 10 min = 72 ± 20 ml versus EDV G1 60 min = 76 ± 19 (p = 0,002); EDV G2 10 min = 98 ± 46 ml versus EDV G2 60 min = 105 ± 38 (p = 0,08); ESVG1 10 min = 22 ± 12 m versus ESV G1 60 min = 25 ± 12 (p = 9E-04); ESVG2 10 min = 47 ± 35 ml versus ESV G2 60 min = 53 ± 36 (p = 3E-04).ConclusionsThis study suggests that early gated Sestamibi SPECT after stress provides same perfusion analysis than classical late imaging.     

Immobilized glucose oxidase on different supports for biotransformation removal of glucose from oligosaccharide mixtures

Four immobilized forms of glucose oxidase (GOD) were used for biotransformation removal of glucose from its mixture with dextran oligosaccharides. GOD was biospecifically bound to Concanavalin A-bead cellulose (GOD-ConA-TBC) and covalently to triazine-bead cellulose (GOD-TBC). Eupergit C and Eupergit CM were used for preparation of other two forms of immobilized GOD: GOD-EupC and GOD-EupCM. GOD-ConA-TBC and GOD-EupC exhibited the best operational and storage stabilities. pH and temperature optima of these two immobilized enzyme forms were broadened and shifted to higher values (pH 7 and 35 °C) in comparison with those of free GOD. The decrease of V_max values after immobilization was observed, from 256.8 ± 7.0 μmol min⁻¹ mg_GOD⁻¹ for free enzyme to 63.8 ± 4.2 μmol min⁻¹ mg_GOD⁻¹ for GOD-ConA-TBC and 45 ± 2.7 μmol min⁻¹ mg_GOD⁻¹ for GOD-EupC, respectively. Depending on the immobilization mode, the immobilized GODs were able to decrease the glucose content in solution to 3.8–15.6% of its initial amount The best glucose conversion, was achieved by an action of GOD-EupCM on a mixture of 100 g dextran with 9 g of glucose (i.e. 98.7% removal of glucose).     

Effects of dietary nitrate supplementation on the oxygen cost of exercise and walking performance in individuals with type 2 diabetes: a randomized,double-blind,placebo-controlled crossover trial

Dietary nitrate supplementation has been shown to reduce the oxygen (O₂) cost of exercise and enhance exercise tolerance in healthy individuals. This study assessed whether similar effects could be observed in individuals with type 2 diabetes (T2DM). In a randomized, double-blind, placebo-controlled crossover study, 48 participants with T2DM supplemented their diet for 4 days with either nitrate-rich beetroot juice (70 ml/day, 6.43 mmol nitrate/day) or nitrate-depleted beetroot juice as placebo (70 ml/day, 0.07 mmol nitrate/day). After each intervention period, resting plasma nitrate and nitrite concentrations were measured subsequent to participants completing moderate-paced walking. Pulmonary gas exchange was measured to assess the O₂ cost of walking. After a rest period, participants performed the 6-min walk test (6MWT). Relative to placebo, beetroot juice resulted in a significant increase in plasma nitrate (placebo, 57±66 vs beetroot, 319±110 µM; P < 0.001) and plasma nitrite concentration (placebo, 680±256 vs beetroot, 1065±607 nM; P < 0.001). There were no differences between placebo juice and beetroot juice for the O₂ cost of walking (946±221 vs 939±223 ml/min, respectively; P = 0.59) and distance covered in the 6MWT (550±83 vs 554±90 m, respectively; P = 0.17). Nitrate supplementation did not affect the O₂ cost of moderate-paced walking or improve performance in the 6MWT. These findings indicate that dietary nitrate supplementation does not modulate the response to exercise in individuals with T2DM.     

Intra- and interday reliability of voluntary and electrically stimulated isometric contractions of the quadriceps femoris

The reliability of voluntary and electrically stimulated isometric contractions of m. quadriceps femoris of male participants (n = 10; age 30 ± 8 years; height 1.79 ± 0.05 m; body mass 79.4 ± 8.3 kg) was investigated using ratio limits of agreement (LoA) on a time scale common to examine recovery from muscle damaging exercise. No systematic changes in reliability occurred over time (baseline versus 2, 24, 48, and 72 h). Maximal voluntary contraction (MVC) and interpolated twitch technique (ITT) showed no mean bias (P > 0.05) with 95% LoA of ±12.7 and ±5.4, respectively. Resting twitch and potentiated doublet peak force showed no mean bias (P > 0.05). However, 95% LoA were smaller for the doublet (±13.9) than the twitch (±32.0). Twitch and doublet rates showed similar trends. Ratio of low (20 Hz) to high (50 Hz) frequency forces showed no mean bias (P > 0.05) and 95% LoA of (±9.2). However, there was significant mean bias (P < 0.05) and wider 95% LoA for peak force, contraction and relaxation parameters of the low and high frequency forces. In conclusion, MVC, ITT, potentiated doublet and the ratio of low to high frequency forces are recommended to most reliably examine functional muscle recovery between 2 and 72 h after damaging exercise.     

Characterization of the N-oxygenase AurF from Streptomyces thioletus

AurF catalyzes the N-oxidation of p-aminobenzoic acid to p-nitrobenzoic acid in the biosynthesis of the antibiotic aureothin. Here we report the characterization of AurF under optimized conditions to explore its potential use in biocatalysis. The pH optimum of the enzyme was established to be 5.5 using phenazine methosulfate (PMS)/NADH as the enzyme mediator system, showing ∼10-fold higher activity than previous reports in literature. Kinetic characterization at optimized conditions give a K_m of 14.7 ± 1.1 μM, a k_cat of 47.5 ± 5.4 min⁻¹ and a k_cat/K_m of 3.2 ± 0.4 μM⁻¹ min⁻¹. PMS/NADH and the native electron transfer proteins showed significant formation of the p-hydroxylaminobenzoic acid intermediate, however H₂O₂ produced mostly p-nitrobenzoic acid. Alanine scanning identified the role of important active site residues. The substrate specificity of AurF was examined and rationalized based on the protein crystal structure. Kinetic studies indicate that the K_m is the main determinant of AurF activity toward alternative substrates.     

Novel l-adenosine analogs as cardioprotective agents

Two l-nucleosides, l-3′-amino-3′-deoxy-N⁶-dimethyladenosine (l-3′-ADMdA) 1, previously synthesized in our laboratory, and the novel l-3′-amino-3′-deoxy-N⁶-methyladenosine-5′-N-methyluronamide (l-3′-AM-MECA) 2 were evaluated in an ischemia/reperfusion model on Langendorff perfused mouse heart. l-3′-ADMdA 1 was found to enhance functional recovery from ischemia (32.2 ± 3.7 cm H₂O/s % rate pressure product, compared to 21.3 ± 1.4 for the control and 30.7 ± 3.4 for adenosine) and increase the time to onset of ischemic contracture (14.5 ± 0.9 min, compared to 10.5 ± 1.0 min for the control and 13.6 ± 0.6 min for adenosine) comparable to adenosine. Consistent with the functional recovery data, decreased infarction area was seen in the case of 1 (19.1 ± 8.4, compared to 40.5 ± 7.2% for the control and 11.5 ± 2.1% for adenosine). In contrast, l-3′-AM-MECA 2 did not show significant functional recovery, increased onset of contracture, nor decreased infarction area compared to control. Unlike adenosine, neither 1 nor 2 induced cardiac standstill in mouse heart.     

The effect of ice-slushy consumption on plasma vasoactive intestinal peptide during prolonged exercise in the heat

The aim of this study was to determine the effect of exercise in the heat on thermoregulatory responses and plasma vasoactive intestinal peptide concentration (VIP) and whether it is modulated by ice-slushy consumption. Ten male participants cycled at 62% $\dot{V}{\mathrm{O}}_2$max for 90 min in 32 °C and 40% relative humidity. A thermoneutral (37 °C) or ice-slushy (−1 °C) sports drink was given at 3.5 ml kg⁻¹ body mass every 15 min during exercise. VIP and rectal temperature increased during exercise (mean±standard deviation: 4.6±4.4 pmol L⁻¹, P=0.005; and 1.3±0.4 °C, P<0.001 respectively) and were moderately associated (r=0.35, P=0.008). While rectal temperature and VIP were not different between trials, ice-slushy significantly reduced heat storage (P=0.010) and skin temperature (time×trial interaction P=0.038). It appears that VIP does not provide the signal linking cold beverage ingestion and lower skin temperature in the heat.