Housing conditions and breed are associated with emotionality and cognitive abilities in riding school horses

Horses’ emotional reactivity is of a major importance in riding schools where a variety of more or less experienced riders are present. Horses’ learning abilities may also be important for work. Previous studies have shown that different intrinsic or extrinsic factors, such as breed, housing conditions, sire, and work may have an influence, and that different facilities present horses with different characteristics. In this study, we tested the reactions of 184 horses from 22 riding schools, all practicing the same type of work, but differing in particular in terms of housing conditions, to 3 emotionality tests and to one instrumental learning task, in order to (1) try and characterize riding schools, and (2) determine how general management could explain some of the potential differences observed between sites.Multivariate analysis conducted on the whole set of data showed that riding schools could be classified into four categories from those where horses showed low emotionality levels and good learning abilities to those where horses showed high emotionality levels and poor learning abilities. Breed, in accordance to previous studies, had an impact on the time to cross a novel obstacle in hand (“Bridge Test”) (Kruskall–Wallis, H (14, N = 184) = 27.08, p < 0.05) while housing conditions (e.g. box housing) influenced emotionality (emotionality index, MW: p < 0.05), when horses were released in the arena. Box housing was associated with more active locomotion patterns (including trot, canter and passage) (MW: p < 0.005). These results underline the importance for riding schools to take into account both the individual characteristics of the horse, and in particular the breed, and to consider the impact of general management including housing conditions, on horses’ reactivity, and its consequences in terms of human security.     

Prevalence of anti-Toxoplasma gondii antibody in domestic horses in Japan

The present study is the first report that investigated the seroprevalence of Toxoplasma gondii infection in domestic horses in various prefectures of Japan and analyzed risk factors for seropositivity. We performed a latex agglutination test for riding/racing horses from 11 prefectures in Japan (783 samples) and 4 groups of Japanese native horses (254 samples). The total seroprevalence of anti-T. gondii antibody in horses examined in this study was 4.24% (44/1037). As for riding/racing horses, we did not find a statistically different T. gondii seroprevalence between sampling prefectures. In contrast, seroprevalence of T. gondii in older horses (> 21 years) was significantly higher than that in younger horses (< 5 years and 11–15 years). There was no significant difference in T. gondii seroprevalence between riding/racing horses and Japanese native horses. Logistical regression analysis revealed that age, but not sex and usage, is a significant risk factor of T. gondii infection for domestic horses in Japan. These findings suggest that domesticated horses in Japan can be horizontally infected with T. gondii by ingestion of food or water contaminated with oocysts.     

Effects of enrichment items on activity and social interactions in domestic horses (Equus caballus)

The aim of this study was to investigate the use of items intended to provide enrichment during turnout, both for individual and group kept horses in an attempt to reduce the amount of passive behaviours. The study was divided into two parts, where study 1 involved eight horses rotated through eight individual paddocks, each containing one of seven enrichment items and one paddock being kept without item, functioning as a control. The horses’ item-directed behaviours; passive behaviours or other non-item related activities were scored using instantaneous sampling, every minute for 1 h at the beginning and the end of the turnout period. Study 2 involved six horse groups (3–6 horses) and the same scoring methods and ethogram as in study 1. The four items that the horses interacted the most with during study 1 (straw STRA, ball filled with concentrates CBALL, branches BRAN and scratching pole POLE) are investigated in study 2. In addition, the amount of social interactions was recorded.Both horses kept individually (P < 0.05) and in groups (P < 0.0001) performed significantly more item-directed behaviours towards edible items like STRA and CBALL than other objects. There was, however, no overall relation between the numbers of item-directed behaviours and the number of passive behaviours observed, indicating that the enrichment items did not alone reduce the amount of passive behaviours during turnout periods. Such a reduction was, however, only apparent when horses spent more time eating green leaves growing on the paddock surface (R = ?0.97 study 1, R = ?0.67 study 2, P < 0.0001). Access to STRA in group kept horses also seemed to reduce the amount of agonistic behaviours (P < 0.0001). In conclusion, if grass is not available in paddocks, the provision of roughage reduces the amount of passive behaviours in singly kept horses and it also reduces the risk of agonistic interactions between horses kept in group.     

Object recognition and generalisation during habituation in horses

The ability of horses to habituate to frightening stimuli greatly increases safety in the horse–human relationship. A recent experiment suggested, however, that habituation to frightening visual stimuli is relatively stimulus-specific in horses and that shape and colour are important factors for object generalisation (Christensen et al., 2008). In a series of experiments, we aimed to further explore the ability of horses (n = 30, 1 and 2-year-old mares) to recognise and generalise between objects during habituation. TEST horses (n = 15) were habituated to a complex object, composed of five simple objects of varying shape and colour, whereas CONTROL horses (n = 15) were habituated to the test arena, but not to the complex object. In the first experiment, we investigated whether TEST horses subsequently reacted less to i) simple objects that were previously part of the complex object (i.e. testing for object recognition) and ii) a novel object (new shape and colour, i.e. testing for object generalisation), compared to CONTROLS. In the second experiment we investigated whether TEST horses reacted to a change in object order and object location. Behavioural reactions to the object, latency to eat, total eating time and heart rate were recorded. Compared to CONTROLS, TEST horses reacted significantly less towards objects, which were previously part of the complex object (e.g. mean heart rate; P = 0.006), indicating object recognition. In contrast to our expectations, TEST horses also reacted significantly less towards the novel object (e.g. mean heart rate; P = 0.018), suggesting that they were capable of object generalisation. We also found that TEST horses showed an increase in exploratory behaviour when objects within the complex object changed order and location (both P < 0.001), whereas there was no increase in heart rate, indicating that the horses were not frightened by the changes. The results demonstrate that it is possible to increase object generalisation in horses by habituating them to a range of colours and shapes simultaneously. This knowledge greatly affects the way in which horses may be trained to react calmly towards frightening objects.     

Tri-axial accelerometer analysis techniques for evaluating functional use of the extremities

Activity monitors provide an objective mechanism for evaluating patient function. It is unclear what similarities or unique information may be yielded using different analyses. Fifteen patients scheduled to undergo shoulder arthroplasty and fifteen matched control subjects wore tri-axial accelerometer activity monitors bilaterally at the lower (wrist) and upper (biceps) arm for 3 days. Measures of central tendency, variance, sample entropy, and asymmetry were calculated. A novel technique to evaluate time distribution of activity intensity was also performed. Within both groups there was a difference in central tendency and variance when comparing dominant and non-dominant limbs for both the lower (Controls: Mean Activity, P < 0.001; Max Activity, P < 0.001; Patients: Mean Activity, P = 0.044; Max Activity, P = 0.009) and upper (Controls: Mean Activity, P < 0.001; Max Activity, P = 0.046; Patients: Mean Activity, P = 0.002; Max Activity, P = 0.049) arm. Within group differences were also present for lower arm entropy in both groups (Controls, P < 0.001; Patients P = 0.041), and at the upper arm for patients (P = 0.003). There were differences between groups for the asymmetry index for both the lower (P = 0.033) and upper arm (P = 0.005), and maximum activity level of the lower arm (P = 0.05). Between group differences were present for time distribution of activity intensity, as the involved upper arm of patients was inactive for a greater time than controls (P = 0.013). These results highlight unique information provided by multiple analysis methods, and include a novel approach of evaluating the distribution of time spent across variable intensity activities.     

Evaluation and comparison of four methods of ranking horses based on reactivity

Four methods of ranking horses on reactivity were evaluated and compared: isolation from conspecifics, presentation of a static novel stimulus, traversing a novel stimulus in a runway (isolation, novel stimulus and runway tests, respectively) and assigning subjective emotionality scores. In all tests, horses’ heart rates were recorded and behaviour was videotaped. To be considered a valid test of reactivity, at least one heart rate and one behavioural measurement in the test had to change significantly between treatments (tranquilizer administation versus sham tranquilizer administration), and behavioural measures had to be displayed in at least 75% of the trials. Forty horses performed each of the three tests daily on three different days in a switchback design. Horses were assigned randomly to a daily test sequence, which was maintained throughout the study. In the runway test, no significant difference in heart rate values in tranquilized and non-tranquilized horses was found, and no behavioural attribute was displayed in more than 52% of the trials; therefore it was rejected as a valid test of reactivity. Both isolation and novel stimulus tests produced valid measurements. Mean heart rate was the most precise physiological measure for these tests, and walking and defecation frequency were the most precise behavioural measures for novel stimulus and isolation tests, respectively. Mean heart rates on the novel stimulus and isolation tests were correlated (r_s = 0.79, P < 0.01) indicating that these tests produced similar rankings based on physiological responses. However, behavioural measures ranked horses differently (r_s = 0.27, P < 0.10) on the tests. Rank correlations between mean heart rates and behavioural measures were higher in the novel stimulus (r_s = 0.66, P < 0.01) than the isolation test (r_s = 0.55, P < 0.01), indicating that the novel stimulus test ranked horses based on either physiological or behavioural responses more similarly than did the isolation test. Therefore, the novel stimulus test was considered the more accurate evaluation of reactivity. Subjective emotionality scores were correlated moderately with mean heart rates (r_s > 0.33, P < 0.01) from the novel stimulus and isolation tests and with walking scores (r_s = 0.47, P < 0.01) from the novel stimulus test. Assignment of subjective emotionality scores was not as accurate as the novel stimulus or isolation tests in ranking horses for reactivity. Using physiological data alone, combining physiological and behavioural measurements or using more than one behavioural measurement in reactivity tests may reflect the reactivity of the horse better than a single behavioural measurement.     

The eccentric,concentric strength relationship of the hamstring muscles in chronic low back pain

ObjectiveThe objective of this study was to measure hamstring muscle eccentric and concentric strength in individuals with and without low back pain (LBP). Two composite scores for the relative balance of eccentric to concentric strength at the different movement velocities were calculated (the DEC and SEC), to determine whether or not self perceived pain, disability, or fear avoidance measures were associated with hamstring strength characteristics.DesignCross-sectional repeated measures design.SettingUniversity laboratory.ParticipantsFifteen individuals with chronic LBP and 15 matched controls.Main outcome measuresIsokinetic eccentric and concentric strength at 30° s^?1 and 120° s^?1_. Composite scores (DEC and SEC) based on peak torque were calculated to evaluate the relationship between the different muscle actions across the test velocities. Self report measures included the Oswestry disability index, general health and well being, fear avoidance, and pain.ResultsEccentric/concentric strength ratio at 30° s^?1 was higher for the LBP group (F(1,58) = 4.81, p = 0.032). The SEC was also higher for the LBP (F(1,58) = 5.97, p = 0.018). Fear avoidance beliefs and mental well-being were significantly associated with the SEC only in the LBP group (adjusted r² = 0.26, (F(2,27) = 5.8, p = .008). For the control group both the DEC and SEC were associated with self report measures. Matched differences between groups’ for the SEC were best explained by fear avoidance beliefs about work (adjusted r² = 0.12, F(1,28) = 5.1, p = 0.03).ConclusionReduced concentric relative to eccentric strength is best identified by the SEC. The SEC was significantly associated with impaired self report measures of fear avoidance and mental well being in individuals with LBP. Differences between groups for the SEC were best explained by fear avoidance beliefs about work.     

Behavioural effects of embryonic exposure to corticosterone in chickens

We tested the hypothesis that exposure of chick embryos to corticosterone leads to increased fear, reduced competitive ability, reduced ability to cross a barrier and reduced growth in juvenile chicks. Behaviour was studied in birds subjected to three different egg injection treatments: a negative control (no treatment of eggs), a positive control (100 μl sesame oil vehicle) and a corticosterone treatment (0.6 μg corticosterone in 100 μl sesame oil). Eggs were injected prior to incubation and the behaviour of chicks was studied during the first 4 weeks of life. Corticosterone treatment increased fear in chicks, as indicated by greater avoidance of an observer in the home pen at 2 weeks of age (P < 0.0001), reduced ability to cross a wall to access feed at 2 weeks of age (P < 0.05) and reduced ability to compete for a wormlike object at 4 weeks of age (P < 0.01). Treatment with corticosterone also reduced body weight at 1 week of age (P < 0.003) and 4 weeks of age (P < 0.04), but not at hatch (P < 0.28). The sesame oil vehicle reduced fear (P < 0.0001), but had no other significant effects. These results indicate that embryonic exposure to corticosterone leads to behavioural and growth deficits in chicks.     

Genome size variation and polyploidy in the resurrection plant genus Ramonda: Cytogeography of living fossils

The genus Ramonda includes three preglacial paleoendemic species surviving as the rare resurrection angiosperms of the Northern hemisphere in refugia habitats in the Balkan (Ramonda nathaliae and Ramonda serbica) and Iberian Peninsulas (Ramonda myconi). This study focuses on: assessing genome size and base composition, determining chromosome number and ploidy level in several populations, evaluating inter- and intra-specific variations in DNA content and chromosome number as well as looking for the possible hybridization in the sympatric zones of Balkan species. R. nathaliae and R. myconi are diploid species (2n = 2x = 48) while R. serbica is hexaploid (2n = 6x = 144). The mean 2C DNA values ranged from 2.30 pg for R. nathaliae to 2.59 pg for R. myconi compared to 7.91 pg for R. serbica. The base composition for R. nathaliae was 42.1% GC, for R. myconi 39.9% and for R. serbica 41.2%. In one population of R. serbica the DNA content ranged from 2C = 7.65 to 11.82 pg, revealing different ploidy levels among its individuals. In sympatric populations genome size was intermediary (～5 pg) between the diploid and hexaploid classes which indicates the hybridization ability between R. serbica and R. nathaliae. It appears that polyploidization is the major evolutionary mechanism in the genus Ramonda.     

Mixing at the beginning of fattening moderates social buffering in beef bulls

Cattle establish strong preferential associations early in their life that determine the cohesiveness within a group and can diminish fear responses (i.e. social buffering). Mixing beef bulls at the beginning of fattening may lead to less cohesiveness. Moreover, mixing beef bulls induces aggressive interactions, especially when the bulls are of similar weights. This study was aimed at evaluating the cohesiveness and fear responses of 64 bulls that were either mixed versus unmixed and of homogeneous vs. heterogeneous body weights (2 × 2 factorial design, four groups of four bulls each per treatment). Social cohesiveness was assessed during food competition and a social separation test. Fear responses were observed during the social separation test and pre-slaughter handling. During food competition, unmixed bulls exchanged more non-agonistic interactions (P < 0.05) and were more tolerant with each other than mixed bulls (time spent eating: P < 0.05). Unmixed bulls displayed less fear responses during physical separation (elimination: P < 0.05; startle: P < 0.01) and were less stressed by pre-slaughter handling (blood cortisol: P < 0.05). Very few effects could be attributed to the homogeneity of weights within groups. It is concluded that mixing bulls at the beginning of fattening reduces the strength of positive social relationships and renders social buffering less effective during subsequent fattening. We recommend maintaining familiar bulls together throughout fattening in order to preserve cohesiveness within groups and for animals to benefit from social buffering during stressful situations.     

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.     

Molecular distinction of C × R hybrid (Coffea congensis × Coffea canephora) from morphologically resembling male parent using rbcL and matK gene sequences

Interspecific C × R hybrid (Coffea congensis × Coffea canephora) in India is cultivated as mixed population with male parent C. canephora as this species is an efficient pollen donor for enhanced yield. But distinction of C × R hybrid from C. canephora in old plantation is difficult due to varying plant sizes of C × R hybrid and often resembles with C. canephora. C × R hybrid cultivated under different agroclimatic conditions show distinct vegetative growth pattern with varying yields. Thus development of DNA marker for identification of C × R hybrid is important for clonal propagation and seed preparation from selective individuals. In this study, two DNA bar coding loci of chloroplast genome (rbcL and matK) of parents, F1 hybrid and its back cross progeny were partially sequenced to identify SNPs as DNA marker for distinction of C × R hybrid from C. canephora. Seven SNPs in the matK gene sequence and three nucleotides in the rbcL gene sequence were identified as DNA markers for the genetic identity of C. congensis. These SNPs were found in F1 and advanced progenies of C × R hybrid due to maternal inheritance. Large number of samples of C × R hybrids with varying morphological features revealed no polymorphism among C × R hybrid and C. congensis. Thus, the SNPs in C. congensis can be used as DNA markers for precise identification of C × R hybrid for production of clones besides tagging the chloroplast inheritance in advanced progenies.     

Measuring voluntary quadriceps activation: Effect of visual feedback and stimulus delivery

Introduction: Quadriceps voluntary activation, assessed via the superimposed burst technique, has been extensively studied in a variety of populations as a measure of quadriceps function. However, a variety of stimulus delivery techniques have been employed, which may influence the level of voluntary activation as calculated via the central activation ratio (CAR). The purpose was to determine the effect of visual feedback, stimulus delivery, and perceived discomfort on maximal voluntary isometric contraction (MVIC) peak torque and the CAR. Methods: Quadriceps CAR was assessed in 14 individuals on two days using three stimulus delivery methods; (1) manual without visual feedback, (2) manual with visual feedback, and (3) automated with visual feedback. Results: MVIC peak torque and the CAR were not different between the automated with visual feedback (MVIC = 3.25, SE = 0.14 N m/kg; CAR = 88.63, SE = 1.75%) and manual with visual feedback (MVIC = 3.26, SE = 0.13 N m/kg, P = 0.859; CAR = 89.06, SE = 1.70%, P = 0.39) stimulus delivery methods. MVIC (2.99, SE = 0.12 N m/kg) and CAR (85.32, SE = 2.10%) were significantly lower using manual without visual feedback compared to manual with visual feedback and automated with visual feedback (CAR P < 0.001; MVIC P < 0.001). Perceived discomfort was lower in the second session (P < 0.05). Conclusion: Utilizing visual feedback ensures participant MVIC, and may provide a more accurate assessment of quadriceps voluntary activation.     

Expression of orexin receptors 1 (OX1R) and 2 (OX2R) in the porcine pituitary during the oestrous cycle

Orexin A and B, also termed hypocretin 1 and 2, are associated with the stimulation of food intake and arousal. The biological actions of the hormones are mediated via two distinct G protein-coupled receptors, termed orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R). OX1R is selective for orexin A and OX2R binds orexin A and orexin B with similar affinity. The present study analyzed mRNA and protein expressions of OX1R and OX2R in adenohypophysis (AP) and neurohypophysis (NP) of cycling pigs. The tissue samples were harvested on days 2–3, 10–12, 14–16, and 17–19 of the oestrous cycle. Using quantitative real-time PCR higher OX1R gene expression was detected in AP on days 2–3 relative to days 10–12, 14–16 and 17–19 (p < 0.05). In NP the OX1R mRNA level was elevated on days 10–12 compared to the remaining stages (p < 0.05). OX2R gene expression in AP was the lowest on days 10–12 (p < 0.05 compared to days 2–3 and 17–19) and the expression peak occurred on days 17–19 (p < 0.05 vs. the all studied stages). In NP the highest (p < 0.05) expression of OX2R mRNA was noted on days 17–19 in relation to the remaining periods. OX1R protein content in AP was greatest on days 10–12 (p < 0.05), whereas in NP it was greatest on days 2–3 and 14–16 (p < 0.05 vs. days 10–12 and 17–19). In both cases the lowest OX1R protein expression was observed during follicular phase (p < 0.05 in relation to three remaining studied stages). OX2R protein in AP was lower (p < 0.05) on days 2–3 and 14–16 compared to days 10–12 and 17–19. In NP the lowest (p < 0.05) expression of this protein was on days 17–19 and the highest on days 10–12 (p < 0.05 compared to days 2–3 and 17–19). In summary, the present findings provide the first evidence that OX1R and OX2R mRNAs and proteins occur in the pituitary of the pig and indicate the dependence of orexin receptor expression on the endocrine reproductive state.     

Insulin-releasing and cytotoxic properties of the frog skin peptide,tigerinin-1R: a structure–activity study

The frog skin host-defense peptide tigerinin-1R (RVCSAIPLPICH.NH₂) is insulinotropic both in vitro and in vivo. This study investigates the effects on insulin release and cytotoxicity of changes in cationicity and hydrophobicity produced by selected substitutions of amino acids by l-arginine, l-lysine and l-tryptophan. The [A5W], [L8W] and [I10W] analogs produced a significant (P < 0.01) increase in the rate of insulin release from BRIN-BD11 rat clonal β cells at concentration of 0.01 nM compared with 0.1 nM for tigerinin-1R. The increase in the rate of insulin release produced by a 3 μM concentration of the [S4R], [H12K], and [I10W] analogs from both BRIN-BD11 cells and mouse islets was significantly greater (P < 0.05) than that produced by tigerinin-1R. No peptide stimulated the release of lactate dehydrogenase at concentrations up to 3 μM indicating that plasma membrane integrity had been preserved. [A5W] tigerinin-1R was the only analog tested that showed cytotoxic activity against human erythrocytes (LC₅₀ = 265 ± 16 μM) and inhibited growth of Escherichia coli (MIC = 500 μM) and Staphylococcus aureus (MIC = 250 μM). The circular dichroism spectra of tigerinin-1R and [A5W] tigerinin-1R indicate that the peptides adopt a mixture of β-sheet, random coil and reverse β-turn conformations in 50% trifluoroethanol/water and methanol/water. Administration of [S4R] tigerinin-1R (75 nmol/kg body weight) to high-fat fed mice with insulin resistance significantly (P < 0.05) enhanced insulin release and improved glucose tolerance over a 60 min period following an intraperitoneal glucose load. The study supports the claim that tigerinin-1R shows potential for development into novel therapeutic agents for treatment of type 2 diabetes mellitus.     

Effect of seat positions on discomfort,muscle activation,pressure distribution and pedal force during cycling

The aim of this study was to measure and analyse discomfort and biomechanics of cycling, i.e., muscle activation, centre of pressure of seat pressure profiles and pedal forces as a function of seat position. Twenty-one recreationally active individuals cycled for 10 min at 100 W on an ergometer cycle using five different seat positions. The neutral position was considered as basic seat position and was compared with upward, downward, forward and backward seat positions. The initial bout was repeated at the end of the recording session. Discomfort increased for upward and backward condition compared with neutral (P < 0.05). Normalized surface electromyography from gastrocnemius decreased in the downward and forward position but increased in the upward and backward position. The minimum force became less negative for forward position compared with neutral seat position (P < 0.05). The degree of variability of centre of pressure increased in the upward and backward position and the entropy of the centre of pressure of sitting posture for backward position decreased compared with neutral seat position (P < 0.05). The present study revealed that consecutive changes of seat position over time lead to increase in discomfort as well as alterations of the biomechanics of cycling.     

Crucial factors affecting the nitrile hydratase production of Mesorhizobium sp. F28

Mesorhizobium sp. F28 contains cobalt-NHase, which effectively converts acrylonitrile into acrylamide. When urea was added to the culture medium, the NHase activity was 62.3 U ml^?1 (R2A–R2A/urea) after 22.5 h of cultivation, which was similar to that in the medium without addition (R2A–R2A, 70.0 U ml^?1). The relative activity of the purified NHase was 100%, 92%, 94%, and 92% in the medium containing, respectively, 0 mM, 2 mM, 5 mM, and 10 mM of urea. Urea had no significant effect on the purified NHase activity of Mesorhizobium sp. F28. This research did not observe the NHase production by Mesorhizobium sp. F28 when acrylonitrile was supplemented in the culture medium except that cobalt ions existed. The highest enzyme activity was 328.5 U ml^?1 as cobalt ions were added in the pre-culture and culture medium after 22.5 h of cultivation (R2A/Co-R2A/Co); compared to media without cobalt ions (R2A–R2A, 22.5 h, 70.5 U ml^?1) this is an almost five-fold enhancement. It can be concluded that culture media containing cobalt ions was beneficial for the formation of active NHase of Mesorhizobium sp. F28.     

The influence of seat height on the mechanical function of the triceps surae muscles during steady-rate cycling

The purpose of this study was to manipulate bicycle seat height in order to perturbate muscle length, contraction velocity and excitation of soleus and medial gastrocnemius muscles. One group of female riders (n = 13) rode a stationary ergometer at 200 W and a cadence of 80 rpm. Individuals rode at a self-selected seat height, a 10% lowered and 5% raised seat position. It was hypothesized that because the two muscles would operate at decreased contraction velocities at the low seat, the integrated EMG would be less for the lowest seat position. The soleus and medial gastrocnemius muscles showed a significant decrease in integrated EMG value with decreased seat height (soleus F_2,24 = 5.4, p < 0.01, gastrocnemius F_2,24 = 51.6, p < 0.0001). The combined effect of the movement at the ankle and knee joints resulted in increased length of gastrocnemius rather than shortening at the lowered seat-height position as anticipated. This suggested that there was a greater role of knee-joint angle in determining the muscle excitation for medial gastrocnemius. The original hypothesis was accepted, confirming the importance of setting proper seat height.     

Improved PCR performance and fidelity of double mutant Neq A523R/N540R DNA polymerase

We previously reported that Neq A523R DNA polymerase is more efficient in PCR than wild-type Neq DNA polymerase, and amplifies products more rapidly. Neq A523R DNA polymerase also amplifies templates more rapidly than Pfu DNA polymerase, but has a lower fidelity than Pfu DNA polymerase. To improve product yield and the fidelity of amplification simultaneously, we constructed and characterized the double mutant Neq A523R/N540R. The yield of PCR products was greater for Neq A523R/N540R DNA polymerase than wild-type and other mutant DNA polymerases, and the Neq double mutant catalyzed amplification of a 12-kb PCR product from a lambda template with an extension time of 3 min. The PCR error rate of Neq A523R/N540R DNA polymerase (6.3 × 10⁻⁵) was roughly similar to that of Pfu DNA polymerase (4.8 × 10⁻⁵), but much lower than those of wild-type Neq DNA polymerase (57.2 × 10⁻⁵), Neq A523R DNA polymerase (13.1 × 10⁻⁵), and Neq N540R DNA polymerase (37.7 × 10⁻⁵). These results indicated that A523R and N540R mutations of Neq DNA polymerase had synergistic effects on its fidelity.     

Bioprocess strategies for mass multiplication of and metabolite synthesis by plant growth promoting pseudomonads for agronomical applications

The exponential substrate feeding (open-loop) and automated feedback substrate feeding (closed loop) strategies were developed to obtain high cell densities of fluorescent pseudomonad strains R62 and R81 and enhanced production of antifungal compound 2,4-diacetylphloroglucinol (DAPG) from glycerol as a sole carbon source. The exponential feeding strategy resulted in increased glycerol accumulation during the fed-batch cultivation when the predetermined specific growth rate (μ) was set at 0.10 or 0.20 h^?1 (<μ_m = 0.29 h^?1). Automated feeding strategies using dissolved oxygen (DO) or pH as feedback signals resulted in minimal to zero accumulation of glycerol for both the strains. In case of DO-based feeding strategy, biomass productivity of 0.24 g/(L h) and 0.29 g/(L h) was obtained for R62 and R81, respectively. Using pH-based feeding strategy, biomass productivity could be increased to a maximum of 0.51 and 0.54 g/(L h), for the strains R62 and R81, respectively, whereas the DAPG concentration was enhanced to 298 mg/L for R62 and 342 mg/L for R81 strains. These yields of DAPG are thus far the highest reported from GRAS organisms.