Validity of the twitch interpolation technique for the assessment of quadriceps neuromuscular asymmetries

This study examined the validity of the twitch interpolation technique for evaluating side-to-side asymmetries in quadriceps neuromuscular function. Fifty-six subjects with a wide range of asymmetries (19 healthy, 24 with unilateral and 13 with bilateral anterior cruciate ligament reconstruction) took part in the study. Supramaximal electrical paired stimuli were delivered to the quadriceps muscle during and immediately after a maximal voluntary contraction (MVC) of the knee extensors (twitch interpolation technique). MVC torque, voluntary activation and resting doublet-evoked torque were measured separately for the two sides, and percent side-to-side asymmetries were calculated for each parameter. MVC torque asymmetry was plotted against voluntary activation asymmetry and doublet-evoked torque asymmetry, and a multiple regression analysis was also conducted. Significant positive correlations were observed between MVC torque asymmetry and both voluntary activation asymmetry (r = 0.40; p = 0.002) and doublet-evoked torque asymmetry (r = 0.53; p < 0.001), and their relative contribution to MVC torque asymmetry was comparable (r = 0.64; p < 0.001). These results establish the validity of the twitch interpolation technique for the assessment of neuromuscular asymmetries. This methodology could provide useful insights into the contribution of some neural and muscular mechanisms that underlie quadriceps strength deficits.     

Neuromuscular fatigue following low-intensity dynamic exercise with externally applied vascular restriction

This study investigated neuromuscular fatigue following low-intensity resistance exercise with vascular restriction (VR) and without vascular restriction (control, CON). Fourteen males participated in two experimental trials (VR and CON) each separated by 48 h. Each participant performed two isometric maximum voluntary contractions (MVCs) before and after five sets of 20 dynamic constant external resistance leg extension exercises (DCER-EX) at 20% of one-repetition maximum (1-RM). The participants were asked to lift (1.5 s) and lower (1.5 s) the load at a constant velocity. Surface electromyography (EMG) was recorded from the vastus lateralis during MVC and DCER-EX. Twitch interpolation was used to assess the percent of maximal voluntary activation (%VA) during the MVC. During performing five sets of 20 DCER-EX, the increases (p < 0.05) in EMG amplitude and decreases (p < 0.05) in EMG mean power frequency were similar for both VR and CON. However, there were significant differences between VR and CON for MVC force, %VA, and potentiated twitch force and significant interactions for EMG amplitude. VR decreased MVC force, %VA, potentiated twitch force, and EMG amplitude more than CON. Our findings suggest that the VR-induced fatigue may have been due to a combination of peripheral (decreases in potentiated twitch) and central (decreases in %VA and EMG amplitude) fatigue.     

Neuromuscular responses to different resistance loading protocols using pneumatic and weight stack devices

The purpose of this study was to examine single repetition characteristics and acute neuromuscular responses to typical hypertrophic (HL), maximal strength (MSL), and power (PL) loadings performed with two of the most common resistance modes; pneumatic and weight stack. Acute responses were assessed by measuring maximal voluntary contraction (MVC), corresponding quadriceps-EMG and resting and superimposed twitch torques. Activation level was calculated from the twitch torques.Decreases in MVC were greater during HL and MSL than during PL. During HL, resting twitch force decreased 8% (P < 0.05) more on the weight stack than on the pneumatic device. Furthermore, loading using the weight stack caused reduced resting twitch force, activation level, and EMG-amplitude after MSL and PL (P < 0.05–0.01).PL on the pneumatic device decreased MVC and rapid force production, while the respective PL on the weight stack device was specific to decreased rapid force production only. However, mean angular velocities and power of the repetitions were higher on the pneumatic device when using light loads.The present study showed that, at least in untrained subjects, the weight stack device induced greater levels of peripheral fatigue during HL. It also led to large central fatigue during MSL and PL. On the other hand, on the pneumatic device contraction velocity with low loads was higher compared to the weight stack device.Therefore, it is recommended that the resistance mode should be chosen according to the specific training goal.     

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.     

Tensiomyography of selected lower-limb muscles in professional soccer players

Tensiomyography is a non-invasive method of neuromuscular assessment used to measure muscle action characteristics, muscle tone, and muscle fiber type, and provides information on acute and chronic responses of muscle to different training loads. The aims of the present study were: to analyse differences in muscle response and mechanical characteristics of two major muscles of the lower extremity in a large group of Spanish soccer players according to playing position, and to provide group norms against which clinical findings may be compared. Data were collected from 78 professional soccer players (age 26.6 ± 4.4 years; height: 179.2 ± 5.3 cm; body mass: 75.8 ± 5.3 kg). Tensiomyography was recorded from the rectus femoris (RF) and biceps femoris (BF) muscles after 2 days without take part in any strenuous exercise or training. Five tensiomyographic parameters were analyzed: maximal displacement (D_m), contraction time (T_c), sustain time (T_s), delay time (T_d), and half-relaxation time (T_r). A good to excellent intra-session reliability was found for all contractile parameters (ICC ranged from 0.78 to 0.95). No significant differences between players of any position were observed in absolute values of BF. However, significant differences were observed for T_c, T_r and T_s between the different playing positions on RF (P < 0.05, effect size ranged from 1.3 to 1.6). Professional soccer players showed muscles with ability to rapidly generate force during contractions. The neuromuscular profile provided could help in identifying the normative data that are important for the different positions in order to optimize the training and recovery process of each individual player.     

Tennis players show a lower coactivation of the elbow antagonist muscles during isokinetic exercises

PurposePrevious studies have suggested that muscle coactivation could be reduced by a recurrent activity (training, daily activities). If this was correct, skilled athletes should show a specific muscle activation pattern with a low level of coactivation of muscles which are typically involved in their discipline. In particular, the aim of this study was to verify the hypothesis that the amount of antagonist activation of biceps brachii (BB) and triceps brachii (TB) is different between tennis players and non-players individuals during maximal isokinetic contractions.MethodsTen young healthy men and eight male tennis players participated in the study. The surface electromyographic signals (sEMG) were recorded from the BB and TB muscles during three maximal voluntary isometric contractions (MVC) of elbow flexors and extensors and a set of three maximal elbow flexions and extensions at 15°, 30°, 60°, 120°, 180° and 240°/s. Normalized root mean square (RMS) of sEMG was calculated as an index of sEMG amplitude.ResultsAntagonist activation (%RMSmax) of TB was significantly lower in tennis players (from 14.0 ± 7.9% at MVC to 16.3 ± 8.9% at 240°/s) with respect to non-players (from 27.7 ± 19.7% at MVC to 38.7 ± 17.6% at 240°/s) at all angular velocities. Contrary to non-players, tennis players did not show any difference in antagonist activation between BB and TB muscles.ConclusionsTennis players, with a constant practice in controlling forces around the elbow joint, learn how to reduce coactivation of muscles involved in the control of this joint. This has been shown by the lower antagonist muscular activity of triceps brachii muscle during isokinetic elbow flexion found in tennis players with respect to non-players.     

Examining the reliability of the flexor carpi radialis V-wave at different levels of muscle contraction

This study examined the reliability and scaling of the flexor carpi radialis (FCR) V-wave during submaximal and maximal voluntary muscle contractions (MVC). 23 participants were tested on three separate sessions. For each session, participants performed isometric wrist flexions at five contraction levels (20, 40, 60, 80 and 100 %MVC). When the target contraction level was reached, a supramaximal electrical stimulus was applied to the median nerve in order to elicit an FCR V-wave. Across all participants, the FCR V-wave amplitude, normalized to its superimposed M-wave amplitude, increased from 0.030 ± 0.001 to 0.143 ± 0.015 (P < 0.001) as the muscle contraction increased from 20 to 100 %MVC. Contraction level did not influence the reliability of evoking the FCR V-wave, as the V-wave demonstrated both stability and consistency. With the exception of a single day main effect during the 20 %MVC condition, V:M_sup was not different across days or trials (P > 0.05) indicating measurement stability. High reliability co-efficients (0.827–0.913) at each contraction level signified measurement consistency. This study establishes that FCR V-waves can be reliably evoked during both submaximal and maximal muscle contractions and suggests the possibility for FCR V-wave recordings to be used to document neuromuscular adaptations associated with factors such as training or fatigue.     

Acute severe male hypo-testosteronemia affects central motor command in humans

Purpose: To indirectly evaluate the effect of androgens on neuromuscular system in humans we analyzed if an induced short-term hypogonadal state (serum total testosterone-TT < 2.3 ng/ml) may affect central drive to skeletal muscle and/or muscle neuro-mechanical performance. Methods: We compared voluntary and electrically evoked muscle sEMG signals from biceps brachii in nine hypogonadal male volunteers (Hypo) and in ten healthy controls (Cont). Serum TT and dihydrotestosterone (DHT) were assayed. Results: With respect to Hypo, Cont exhibited significantly higher median frequency content (MDF) at any angular velocity; normalized MDF [95.9% (SD = 23.3) vs 73.8% (SD = 9.3)]; muscle fiber conduction velocity (CV) from lowest to highest angular velocities; initial MDF at fatigue test [91.78 Hz (SD = 22.03) vs 70.94 Hz (SD = 11.06)] as well as was the normalized slope [−0.64 (SD = 0.14 vs −0.5 (SD = 0.11)]. In the non-fatigued state, Hypo showed a slower single twitches time to peak (TTP). In Cont, half relaxation time (HRT) decreased after fatigue while increased in Hypo (p < 0.05 between groups). A significant correlation between both TT and dihydrotestosterone with MDF and CV was found during voluntary contractions only. Conclusions: A brief exposure to very low serum TT concentration in males seem to determine a reduced excitability of the NM system which, in turn, would favor a predominant recruitment of slow twitch MUs.     

The influence of myosin heavy chain isoform content on mechanical behavior of the vastus lateralis in vivo

This study examined correlations between type I percent myosin heavy chain isoform content (%MHC) and mechanomyographic amplitude (MMG_RMS) during isometric muscle actions. Fifteen (age = 21.63 ± 2.39) participants performed 40% and 70% maximal voluntary contractions (MVC) of the leg extensors that included increasing, steady force, and decreasing segments. Muscle biopsies were collected and MMG was recorded from the vastus lateralis. Linear regressions were fit to the natural-log transformed MMG_RMS–force relationships (increasing and decreasing segments) and MMG_RMS was selected at the targeted force level during the steady force segment. Correlations were calculated among type I%MHC and the b (slopes) terms from the MMG_RMS–force relationships and MMG_RMS at the targeted force. For the 40% MVC, correlations were significant (P < 0.02) between type I%MHC and the b terms from the increasing (r = −0.804) and decreasing (r = −0.568) segments, and MMG_RMS from the steady force segment (r = −0.606). Type I%MHC was only correlated with MMG_RMS during the steady force segment (P = 0.044, r = −0.525) during the 70% MVC. Higher type I%MHC reduced acceleration in MMG_RMS (b terms) during the 40% MVC and the amplitude during the steady force segments. The surface MMG signal recorded during a moderate intensity contraction provided insight on the contractile properties of the VL in vivo.     

Strength training,but not endurance training,reduces motor unit discharge rate variability

This study evaluates and compares the effects of strength and endurance training on motor unit discharge rate variability and force steadiness of knee extensor muscles. Thirty sedentary healthy men (age, 26.0 ± 3.8 yrs) were randomly assigned to strength training, endurance training or a control group. Conventional endurance and strength training was performed 3 days per week, over a period of 6 weeks. Maximum voluntary contraction (MVC), time to task failure (at 30% MVC), coefficient of variation (CoV) of force and of the discharges rates of motor units from the vastus medialis obliquus and vastus lateralis were determined as subjects performed 20% and 30% MVC knee extension contractions before and after training. CoV of motor unit discharges rates was significantly reduced for both muscles following strength training (P < 0.001), but did not change in the endurance (P = 0.875) or control group (P = 0.995). CoV of force was reduced after the strength training intervention only (P < 0.01). Strength training, but not endurance training, reduces motor unit discharge rate variability and enhances force steadiness of the knee extensors. These results provide new insights into the neuromuscular adaptations that occur with different training methods.     

The use of a mirror reduces isolation stress in horses being transported by trailer

Horse trailers are a common form of transportation for horses and ponies and often require the animal to travel alone or with a single companion. The current study investigated the effect of transporting horses alone, in company or with an acrylic safety mirror (measuring 81 cm × 61.5 cm) that provided surrogate companionship. The behavioural and physiological responses of 12 mature horses during a 30-min journey by trailer under the three treatments were compared. Behaviours (vocalisation, eating, head-tossing, pawing, and head-turning) were recorded. In order to assess circulatory changes that occur as part of the response to transport, heart rate (HR), rectal (T_r) and ear-pinna (T_p) temperatures were recorded. When travelling with a live companion significantly less time was spent vocalising (p < 0.001), head-turning (p < 0.001), head-tossing (p < 0.01) and pawing (p < 0.01); eating behaviour increased (p < 0.05). Physiological responses (increases in HR and T_r and decreases in T_p) were also significantly reduced when travelling with a live companion (p < 0.01). Travelling with the mirror did not significantly affect physiological responses compared with travelling alone, but the rise in T_r and fall in T_p was reduced (p = 0.052 and p = 0.051, respectively) and can be considered a trend. When travelling with a mirror significantly less time was spent turning the head (p < 0.01), vocalising (p < 0.05) and head-tossing (p < 0.05); eating behaviour increased (p < 0.05). The only significant difference between travelling with a live companion and a mirror was that the time spent turning the head round was less with a live companion (p < 0.05). The provision of surrogate companionship in the form of a mirror was found to be preferable to travel alone, but where possible a live companion is recommended. Isolation during transportation was found to suppress feeding behaviour. Although peripheral blood flow (T_p) has been used to assess transport stress in other species it has not previously been used in the horse. Further evaluation of this non-invasive measure is now required.     

Acute effects of direct inhibitory pressure over the biceps brachii myotendinous junction on skeletal muscle activation and force output

Force (F) reduction is reported with myotendinous junction (MTJ) manipulation. Autogenic inhibition reflex (AIR) activation is supposed to be the main mechanism. Still, its role remains unclear. The study aimed at assessing the effects of MTJ direct inhibitory pressure (DIP) on neuromuscular activation and F in the elbow flexor (agonist) and extensor (antagonist) muscles. After maximum voluntary contraction (MVC) assessment, thirty-five participants randomly performed submaximal contractions at 20, 40, 60, and 80% MVC. Electromyographic (EMG), mechanomyographic (MMG), and F signals were recorded. Protocol was repeated under (i) DIP (10-s pressure on the biceps brachii MTJ) with the elbow at 120° (DIP₁₂₀), (ii) DIP with the elbow at 180° (DIP₁₈₀), and (iii) without DIP (Ctrl). Electromechanical delay (EMD) components, EMG and MMG root mean square (RMS), and rate of force development (RFD) were calculated. Independently from the angle, DIP induced decrements in MVC, RFD, and RMS of EMG and MMG signals and lengthened the EMD components in agonist muscles (P < 0.05). The DIP-induced decrease in F output of the agonist muscles seems to be possibly due to a concomitant impairment of the neuromuscular activation and a transient decrease in stiffness. After DIP, the antagonist muscle displayed no changes; therefore, the intervention of AIR remains questionable.     

Myoelectric manifestations of fatigue in vastus lateralis,medialis obliquus and medialis longus muscles

The purpose of this study was to determine whether surface electromyography (EMG) assessment of myoelectric manifestations of muscle fatigue is capable of detecting differences between the vastus lateralis and medialis muscles which are consistent with the results of previous biopsy studies. Surface EMG signals were recorded from the vastus medialis longus (VML), vastus medialis obliquus (VMO) and vastus lateralis (VL) muscles during isometric knee extension contractions at 60% and 80% of the maximum voluntary contraction (MVC) for 10 s and 60 s, respectively. Initial values and rate of change of mean frequency (MNF), average rectified value (ARV) and conduction velocity (CV) of the EMG signal were calculated. Comparisons between the two force levels revealed that the initial values of MNF for the VL muscle were greater at 80% MVC compared to 60% MVC (P < 0.01). Comparisons between the vasti muscles demonstrated lower initial values of CV for VMO compared to VL at 60% MVC (P < 0.01) and lower than VML and VL at 80% MVC (P < 0.01). In addition, initial values of MNF were higher for VL with respect to both VML and VMO at 80% MVC (P < 0.01) and initial estimates of ARV were higher for VMO compared to VML at both force levels (P < 0.01 at 60% MVC and P < 0.05 at 80% MVC). For the sustained contraction at 80% MVC, VL demonstrated a greater decrease in CV over time compared to VMO (P < 0.05).These findings suggest that surface EMG signals and their time course during sustained isometric contractions may be useful to non-invasively describe functional differences between the vasti muscles.     

A Comparison of Twice-Daily Biphasic Insulin Aspart 70/30 and Once-Daily Insulin Glargine in Persons With Type 2 Diabetes Mellitus Inadequately Controlled on Basal Insulin and Oral Therapy: A Randomized,Open-Label Study

ObjectiveTo compare efficacy and safety of biphasic insulin aspart 70/30 (BIAsp 30) with insulin (glargine) in type 2 diabetic patients who were not maintaining glycemic control on basal insulin and oral antidiabetic drugs.MethodsIn a 24-week, open-label, parallel-group trial, type 2 diabetic patients who were not maintaining glycemic control on basal insulin (glargine or neutral protamine Hagedorn) + oral antidiabetic drugs were randomly assigned to twice-daily BIAsp 30 + metformin or oncedaily glargine + metformin + secretagogues (secretagogues were discontinued in the BIAsp 30 arm).ResultsOne hundred thirty-seven patients were randomly assigned to the BIAsp 30 group and 143 patients were randomly assigned to the glargine group. Of 280 patients randomized, 229 (81.8%) completed the study. End-of-trial hemoglobin A_1c reductions were − 1.3% (BIAsp 30) vs − 1.2% (glargine) (treatment difference: 95% confidence interval, − 0.06 [− 0.32 to 0.20]; P = .657). Of patients taking BIAsp 30, 27.3% reached a hemoglobin A_1c level < 7.0% compared with 22.0% of patients taking glargine (treatment difference: P = .388). Glucose increment averaged over 3 meals was lower in the BIAsp 30 arm (treatment difference: − 17.8 mg/dL, P = .001). Fasting plasma glucose reductions from baseline were − 13.8 mg/ dL (BIAsp 30) vs − 42.5 mg/dL (glargine) (P = .0002). Final minor hypoglycemia rate, insulin dose, and weight change were higher in the BIAsp 30 arm (6.5 vs 3.4 events/patient per year, P <.05; 1.19 vs 0.63 U/kg; and 3.1 vs 1.4 kg, P = .0004, respectively).ConclusionsDespite not receiving secretagogues, patients taking BIAsp 30 + metformin achieved similar hemoglobin A_1c levels and lower postprandial plasma glucose compared with those receiving glargine + metformin + secretagogues. The large improvement in the glargine group suggests the patients were not true basal failures at randomization. While switching to BIAsp 30 improves glycemic control in this patient population, remaining on basal insulin and optimizing the dose may be equally effective in the short term. (Endocr Pract. 2011;17:41-50)     

Innervation zone shift at different levels of isometric contraction in the biceps brachii muscle

Experiments were carried out to examine whether innervation zone (IZ) location remains stable at different levels of isometric contraction in the biceps brachii muscle (BB), and to determine how the proximity of the IZ affects common surface electromyography (sEMG) parameters. Twelve subjects performed maximal (MVC) and submaximal voluntary isometric contractions at 10%, 20%, 30%, 40%, 50% and 75% of MVC. sEMG signals were recorded with a 13 rows × 5 columns grid of electrodes from the short head of BB. The IZ shifted in the proximal direction by up to 2.4 cm, depending upon the subject and electrode column. The mean shift of all the columns was 0.6 ± 0.4 cm (10% vs. 100% MVC, P < 0.001). This shift biased the average values of mean frequency (+21.8 ± 9.9 Hz, P < 0.001), root mean square (?0.16 ± 0.15 mV, P < 0.05) and conduction velocity (?1.15 ± 0.93 m/s, P < 0.01) in the channels immediately proximal to the IZ. The shift in IZ could be explained by shortening of the muscle fibers, and thus lengthening of the (distal) tendon due to increasing force. These results underline the importance of individual investigation of IZ locations before the placement of sEMG electrodes, even in isometric contractions.     

Acute effects of a dopamine/norepinephrine reuptake inhibitor on neuromuscular performance following self-paced exercise in cool and hot environments

Dopamine/norepinephrine (DA/NE) reuptake inhibitors have been used to manipulate the central mechanisms affecting arousal and motivation during exercise. Eight healthy, physically active males performed 30 min fixed-intensity cycling at 50% W_max followed by 30 min of self paced time trial (TT) with each section interspersed with a 30 s maximal sprint at 9, 19 and 29 min. The DA/NE re-uptake inhibitor administered was bupropion (BUP) versus a placebo (PLA) in either warm (32 °C, BUP32 or PLA32) or moderate (20 °C; BUP20, PLA20) ambient conditions. Core and skin temperature, heart rate and perceptual responses, neuromuscular and hormonal measures were assessed at multiple times throughout the trials and post exercise. Time trial performance remained unchanged across conditions (12.7–13.1 km) although core temperature was elevated in the fixed intensity section of the trials for BUP32 and BUP20 but continued to rise only in BUP32 during the time trial reaching 38.6 °C (P<0.05). NE increased in all conditions from pre-exercise with BUP32 values peaking at the end of TT to 1245.3±203.1 pg/mL (P<0.05) compared to the other conditions. Neuromuscular responses were similar among conditions although peak force was significantly reduced from pre (262±31 N) to post (202±31 N, P<0.05) exercise along with contraction duration (22%, P<0.05) in BUP20. We conclude that DA/NE re-uptake inhibitors influenced thermoregulation in the heat but not exercise performance. DA/NE re-uptake inhibitors are likely to act centrally to override the inhibitory signals for the cessation of exercise with these drugs acting peripherally to reduce the twitch characteristics of skeletal muscle in cooler conditions.     

The effect of time left alone at home on dog welfare

The aim of this study was to investigate the effect of time left alone on dog behaviour and cardiac activity. Twelve privately owned dogs, with no history of separation related behaviour problems, were video-recorded on three different occasions when left alone in their home environment. The treatments lasted for 0.5 h (T_0.5); 2 h (T₂) and 4 h (T₄). Video-recording started 10 min before the owner left the house and continued until 10 min after the owner returned, so that interactions between dog and owner as well as behaviour during separation could be studied. Data on heart rate (HR) and heart rate variability (HRV) were collected within the same time period in each treatment. In addition to analysing behaviours separately, behaviours were also grouped together and defined as new variables; physically active, attentive behaviour, vocal, interaction initiated by owner and interaction initiated by dog. There were no differences in behaviour between treatments at equivalent time intervals until the owner returned, although a number of differences were observed at reunion with the owner. Dogs showed a higher frequency of physical activity (P < 0.05) and attentive behaviour (P < 0.01) in T₂ (0.37 ± 0.07; 0.52 ± 0.08, mean frequency of occurrence/15 s ± SE) and T₄ (0.48 ± 0.08; 0.48 ± 0.07) compared to T_0.5 (0.20 ± 0.07; 0.21 ± 0.05). They also showed more tail wagging (P < 0.01) and interacted more with their owners (P < 0.01) in T₂ (0.27 ± 0.08; 0.47 ± 0.09) and T₄ (0.26 ± 0.04; 0.42 ± 0.09) compared to T_0.5 (0.09 ± 0.04; 0.14 ± 0.03). After a longer time of separation, the dogs also showed higher frequencies of lip licking (P < 0.05) and body shaking (P < 0.05) at the owner's return (T_0.5 = 0.09 ± 0.05; T₂ = 0.24 ± 0.08; T₄ = 0.27 ± 0.06 and T_0.5 = 0.03 ± 0.01; T₂ = 0.08 ± 0.03; T₄ = 0.07 ± 0.01, respectively). There was a tendency for higher HR (P < 0.1) during the first and second minute after reunion in T₂ (127.6 ± 1.25, mean bpm ± SE; 111.3 ± 1.24) compared to T_0.5 (106.2 ± 1.06; 87.5 ± 1.02). According to the results of this study, the effect of time left alone was shown by a more intense greeting behaviour by the dog towards their owner as well as by a higher frequency of physical activity and attentive behaviour when the owner returned, already after 2 h of separation. Although this study cannot distinguish between whether dogs were aware of the length of time they were alone (but did not signal it) or whether they were unaware until reminded of it by the return of their owner, it does confirm that dogs are affected by the duration of time at home alone.     

Plasma trace elements levels are not altered by submaximal exercise intensities in well-trained endurance euhydrated athletes

The purpose of this study was to assess the effect of relative exercise intensity on various plasma trace elements in euhydrated endurance athletes.Twenty-seven well-trained endurance athletes performed a cycloergometer test: after a warm-up of 10 min at 2.0 W kg⁻¹, workload increased by 0.5 W kg⁻¹ every 10 min until exhaustion. Oxygen uptake, blood lactate concentration ([La⁻]_b), and plasma ions (Zn, Se, Mn and Co) were measured at rest, at the end of each stage, and 3, 5 and 7 min post-exercise. Urine specific gravity (U_SG) was measured before and after the test, and subjects drank water ad libitum. Fat oxidation (FAT_OXR), carbohydrate oxidation (CHO_OXR), energy expenditure from fat (EE_FAT), from carbohydrates (EE_CHO) and total EE (EE_T) were estimated using stoichiometric equations. A repeated measure (ANOVA) was used to compare plasma ion levels at each exercise intensity level. The significance level was set at P < 0.05.No significant differences were found in U_SG between, before, and after the test (1.014 ± 0.004 vs. 1.014 ± 0.004 g cm⁻³) or in any plasma ion level as a function of intensity. There were weak significant correlations of Zn (r = 0.332, P < 0.001) and Se (r = 0.242, P < 0.01) with [La⁻]_b, but no relationships were established between [La⁻]_b, VO₂, FAT_OXR, CHO_OXR, EE_FAT, EE_CHO, or EE_T and plasma ion levels.Acute exercise at different submaximal intensities in euhydrated well-trained endurance athletes does not provoke a change in plasma trace element levels, suggesting that plasma volume plays an important role in the homeostasis of these elements during exercise.     

Copper,zinc superoxide dismutase of Curcuma aromatica is a kinetically stable protein

This study details on cloning and characterization of Cu,Zn superoxide dismutase (Ca–Cu,Zn SOD) from a medicinally important plant species Curcuma aromatica. Ca–Cu,Zn SOD was 692 bp with an open reading frame of 459 bp. Expression of the gene in Escherichia coli cells followed by purification yielded the enzyme with K_m of 0.047 ± 0.008 μM and V_max of 1250 ± 24 units/mg of protein. The enzyme functioned (i) across a temperature range of −10 to +80 °C with temperature optima at 20 °C; and (ii) at pH range of 6–9 with optimum activity at pH 7.8. Ca–Cu,Zn SOD retained 50% of the maximum activity after autoclaving, and was stable at a wide storage pH ranging from 3 to 10. The enzyme tolerated varying concentrations of denaturating agent, reductants, inhibitors, trypsin, was fairly resistant to inactivation at 80 °C for 180 min (k_d, 6.54 ± 0.17 × 10⁻³ min⁻¹; t_1/2, 106.07 ± 2.68 min), and had midpoint of thermal transition (T_m) of 70.45 °C. The results suggested Ca–Cu,Zn SOD to be a kinetically stable protein that could be used for various industrial applications.