Functional nickel-based deposits synthesized by focused beam induced processing.

Functional nanostructures fabricated by focused electron/ion beam induced processing (FEBIP/FIBIP) open a promising route for applications in nanoelectronics. Such developments rely on the exploration of new advanced materials. We report here the successful fabrication of nickel-based deposits by FEBIP/FIBIP using bis(methyl cyclopentadienyl)nickel as a precursor. In particular, binary compounds such as nickel oxide (NiO) are synthesized by using an in situ two-step process at room temperature. By this method, as-grown Ni deposits transform into homogeneous NiO deposits using focused electron beam irradiation under O2 flux. This procedure is effective in producing highly pure NiO deposits with resistivity of 2000 Ωcm and a polycrystalline structure with face-centred cubic lattice and grains of 5 nm. We demonstrate that systems based on NiO deposits displaying resistance switching and an exchange-bias effect could be grown by FEBIP using optimized parameters. Our results provide a breakthrough towards using these techniques for the fabrication of functional nanodevices.

Overtrained horses alter their resting pulsatile growth hormone secretion.

The influence of intensified and reduced training on nocturnal growth hormone (GH) secretion and elimination dynamics was studied in young (1.5 yr) Standardbred geldings to detect potential markers indicative for early overtraining. Ten horses trained on a treadmill for 32 wk in age-, breed-, and gender-matched fixed pairs. Training was divided into four phases (4, 18, 6, and 4 wk, respectively): 1) habituation to high-speed treadmill trotting, 2) normal training, in which speed and duration of training sessions were gradually increased, 3) in this phase, the horses were divided into 2 groups: control (C) and intensified trained (IT) group. In IT, training intensity, duration, and frequency were further increased, whereas in control these remained unaltered, and 4) reduced training (RT). At the end of phases 2, 3, and 4, blood was sampled overnight every 5 min for 8 h for assessment of GH secretory dynamics using pulse detection, deconvolution analysis, and approximate entropy (ApEn). Intensified training induced overtraining (performance decreased by 19% compared with C), which was associated with an increase in concentration peaks number (3.6 vs. 2.0, respectively), a smaller peak secretion pattern with a prolonged half-life (15.2 vs. 7.3 min, respectively), and an increased ApEn (0.89 vs. 0.49, respectively). RT did not lead to full recovery for the overtrained horses. The increased irregularity of nocturnal GH pulsatility pattern is indicative of a loss of coordinated control of GH regulation. Longer phases of somatostatin withdrawal are hypothesized to be the underlying mechanism for the observed changes in GH pulsatility pattern.

Normal function of the hypothalamic-pituitary growth axis in three dwarf Friesian foals.

Serial blood samples were collected from three dwarf Friesian foals to examine their endogenous growth hormone (GH) profiles, and the integrity of the GH-insulin-like growth factor-1 (IGF-1) axis was tested in one of them by examining its responses to the administration of GH-releasing hormone (GHRH) and to 10 days of treatment with recombinant equine GH. The basal serum concentrations of IGF-1 in the three dwarf foals were compared with those in nine age-matched normal foals. All the dwarf foals secreted endogenous GH. Stimulation with 7.0 microg/kg GHRH led to a 1400 per cent increase in plasma GH concentration in the dwarf foal tested, and 10 daily subcutaneous treatments with 20 microg/kg recombinant equine GH led to a 100 per cent increase in its serum IGF-1 concentration. The basal serum concentrations of IGF-1 in the dwarf foals were not significantly different from those of the normal foals.

Effects of exogenous insulin on luteolysis and reproductive cyclicity in the mare.

Insulin is a pancreatic hormone that classically regulates carbohydrate and fat metabolism, but also appears to play a role in various reproductive processes. A preliminary study suggested insulin production by day 10 to 18 equine conceptuses. The aim of the present study was to examine the hypothesis that insulin is the conceptus signal responsible for maternal recognition of pregnancy (MRP) in the mare, or otherwise influences reproductive cyclicity during the MRP period. Six Warmblood mares were treated daily during days 7 to 17 after ovulation of two successive oestrous cycles with either (short and intermediate acting) insulin or control saline. Mares were assigned randomly to treatment, and crossed over during the subsequent cycle. Time of ovulation and corpus luteum surface area were determined by serial transrectal ultrasonographic examination of the mares' ovaries, and daily jugular vein blood samples were analysed for progesterone and luteinizing hormone (LH) concentrations. On day 14 of dioestrus, the luteolytic drive was examined by measuring systemic 15-ketodihydroprostaglandin F(2 alpha) (PG-metabolite) release in response to oxytocin challenge. In addition, yolk sac fluid recovered from 32 day 10 to 14 equine conceptuses was analysed for insulin concentrations. Insulin administration did not affect luteal size, dioestrus length, the interovulatory interval, or circulating LH concentrations. Insulin administration also failed to suppress oxytocin-induced PGF(2 alpha) release, and tended to depress systemic progesterone concentrations. Finally, insulin could not be detected in the yolk sac fluid of day 10 to 14 equine conceptuses by radio-immunoassay. It is concluded that insulin administered daily during days 7 to 17 of dioestrus has little or no effect on reproductive cyclicity in the mare, and is unlikely to be the MRP signal.

Ultrasonographic measurements of localized fat accumulation in Shetland pony mares fed a normal v. a high energy diet for 2 years.

Health risks associated with obesity are more likely a factor of the localization of fat excess, rather than of elevated BW per se. The aim of this randomized controlled clinical trial was to determine the effect of a long-term high energy diet on BW, fat accumulation and localization. Eight Shetland pony mares, 3 to 7 years old, were randomly divided into a control and a high energy (HE) diet group fed either maintenance or double maintenance energy requirements (200% net energy (NE)) for two consecutive summers, with a low energy diet in the winter in between. Body condition score (BCS) did not differ between the groups at the onset of the study (control 5.6±0.75 v. HE 6.3±0.5). From 12 weeks after starting the diet, ultrasonography of five different locations (retroperitoneal, axillary, withers, intercostal and rump) for adipose deposition, BCS and BW were measured monthly during the period that ponies received different diets. Statistical analysis was performed using a linear mixed-effects model with post hoc Bonferroni testing. P values <0.05 were considered significant. At week 12 after the onset of the diet, fat thickness in the HE group was significantly greater than in the control group. During the monitoring period, the HE group showed a significant increase in mean (±SE) BW (+52%, 265±13.94kg) and BCS (+70%; to 9.0±0.4), while the control group was unchanged (BW 160±13.98 kg; BCS 3.8±0.4). At all locations, the fat depth in the HE group increased significantly, with the highest increase noted for retroperitoneal deposits. The conclusions were that a 200% NE diet induced subcutaneous and retroperitoneal fat accumulation, with the greatest increase in intra-abdominal deposits. The moderate increase of the subcutaneous fat depth followed by a plateau phase suggests the existence of a limit of adipose tissue expandability, as in man.

Mouldable, thermoplastic, glue-on frog-supportive shoes change hoof kinetics in normal and obese Shetland ponies.

BACKGROUND: Obesity and hyperinsulinaemia are frequently encountered in the equine population and risk factors for the development of laminitis. There are many options for hoof support that claim a beneficial effect, but often the scientific evidence is scarce. OBJECTIVES: To quantify the effect of frog-supportive shoes on hoof kinetics in normal and obese ponies. STUDY DESIGN: Controlled in vivo trial. METHODS: Ten Shetland mares (n = 10) with a normal (n = 5) or obese (n = 5) body condition were led over a dynamically calibrated pressure plate before (T0), immediately after (T1) and 72 h (T2) after application of the shoes. The following locomotor variables were measured: stance duration (StDur), vertical impulse (VI), peak vertical force (PVF), time to PVF and time from PVF to lift off. The hoof print was divided into a toe and heel region and the StDur toe-heel index was calculated. The toe-heel hoof balance curves of the vertical force were plotted throughout the stance phase. RESULTS: The VI and PVF increased significantly 72 h after application of the shoes, when compared with T0 and T1. The StDur toe-heel index and toe-heel balance curves were significantly different between the normal and obese ponies. These variables became more comparable between the groups after application of the frog-supportive shoes. MAIN LIMITATIONS: It would have been interesting to measure the effect of the shoe in patients with acute laminitis. However, this would have had major welfare implications. CONCLUSIONS: The obese ponies moved more carefully than the normal group, demonstrated by a decreased loading of the toe area. The data illustrate that the ponies became more comfortable 72 h after application of the shoes, with a pronounced effect in the obese group. Thus, these results suggest that frog-supportive shoes could be beneficial, especially for obese ponies.

Effect of exercise on apparent total tract digestibility of nutrients and faecal recovery of ADL and TiO2 in ponies.

Exercise and physical training are known to affect gastrointestinal function and digestibility in horses and can lead to inaccurate estimates of nutrient and energy digestibility when markers are used. The effect of exercise on apparent nutrient digestibility and faecal recoveries of ADL and TiO2 was studied in six Welsh pony geldings subjected to either a low- (LI) or high-intensity (HI) exercise regime according to a cross-over design. Ponies performing LI exercise were walked once per day for 45 min in a horse walker (5 km/h) for 47 consecutive days. Ponies submitted to HI exercise were gradually trained for the same 47 days according a standardized protocol. Throughout the experiment, the ponies received a fixed level of feed and the daily rations consisted of 4.7 kg DM of grass hay and 0.95 kg DM of concentrate. The diet was supplemented with minerals, vitamins and TiO2 (3.0 g Ti/day). Total tract digestibility of DM, organic matter (OM), CP, crude fat, NDF, ADF, starch, sugar and energy was determined with the total faeces collection (TFC) method. In addition, DM and OM digestibility was estimated using internal ADL and the externally supplemented Ti as markers. Urine was collected on the final 2 days of each experimental period. Exercise did not affect apparent digestibility of CP, crude fat, starch and sugar. Digestibility of DM (DMD), OM (OMD), ADF and NDF tended to be lower and DE was decreased when ponies received the HI exercise regime. For all treatments combined, mean faecal recoveries of ADL and Ti were 87.8±1.7% and 99.3±1.7%, respectively. Ti was not detected in the urine, indicating that intestinal integrity was maintained with exercise. Dry matter digestibility estimated with the TFC, ADL and Ti for ponies subjected to LI exercise were 66.3%, 60.3% and 64.8%, respectively, while DMD for HI ponies were 64.2%, 60.3% and 65.2%, respectively. In conclusion, physical exercise has an influence on the GE digestibility of the feed in ponies provided with equivalent levels of feed intake. In addition, the two markers used for estimating apparent DMD and OMD indicate that externally supplemented Ti is a suitable marker to determine digestibility of nutrients in horses performing exercise unlike dietary ADL.

Hormonal responses to acute exercise, training and overtraining. A review with emphasis on the horse.

Overtraining is an imbalance between training and recovery leading to symptoms associated with a neuroendocrine dysbalance called the overtraining syndrome, a disease characterized by behavioral, emotional and physical symptoms similar with depression. Although the prevalence of overtraining is high in human and equine athletes, at present no sensitive and specific test is available to prevent or diagnose overtraining. Nowadays, it is believed that combination of different (hormonal) parameters appear to be the best indicators of overtraining. Therefore, this review provides a summary of previous literature examining the response of the hypothalamic-pituitary-adrenal (HPA) axis and the growth hormone-insulin-like growth factor-I (GH-IGF-I) axis to acute and chronic exercise as well as overtraining in humans and horses. The exercise induced hormonal responses seem to be equal for the equine as well as the human athlete, which makes comparisons possible. Repeated bouts of exercise are suggested to provide a way to detect subtle changes in hormonal responses in the individual athlete, which may make them an important tool in detecting early overtraining. This should be combined with corticotropin releasing hormone (CRH) stimulation tests and basal ACTH and GH pulsatility determination. Further research is needed to establish the correct training intensity and rest period for the exercise test in equines.

Risk factors associated with health disorders in sport and leisure horses in the Netherlands.

Horses are used for a wide variety of purposes from being used for recreational purposes to competing at an international level. With these different uses, horses have to adapt to numerous challenges and changes in their environment, which can be a challenge itself in continuously safeguarding their welfare. The objective of this study was to assess the prevalence of health disorders with clinical examination and identify possible risk factors of health disorders affecting horse welfare in professional husbandry systems in the Netherlands. With the use of fixed protocols for recording health aspects in horses, 150 horse farms voluntarily participating in the study were assessed by trained assessors. On each farm, 20 horses were clinically examined, in total almost 3,000 animals. This study recorded on the basis of the clinical examinations: the respiratory system (i.e., abnormal breathing [1%], coughing [1%], nasal discharge [1.9%]), body condition (i.e., 18.8% fat body condition and 6.4% poor body condition), locomotion (14.5% exhibited irregularity of locomotion and 4.8% were lame), back palpation (a light response [22.6%] and moderate to severe response [8.4%]), mouth (i.e., irregularities on mouth corners [3.4%] and bars [3.4%]), and ocular discharge (12%). Risk factor analysis, stepwise using mixed model regression, demonstrated several risk factors for health aspects. Horses used for instruction (riding lessons) were almost two times more at risk to develop moderate to severe back pain compared to horses used for recreation (odds ratios [OR] = 0.54) or for competition (OR = 0.61). Horses used for instruction (riding school lessons), breeding, or recreation all had a higher risk for irregular locomotion or lameness compared to competition horses (OR = 0.42, OR = 0.55, OR = 2.14, respectively). Horses used for recreation were more prone to have a higher BCS compared to horses used for breeding (OR = 3.07) and instruction (OR = 2.06). The prevalence of health problems and the identified risk factors are valid for the horses in the present study in which farms voluntarily participated. Furthermore, the results may provide the basis for horse welfare and health programs on farm and horse industry levels. With the development of a valid welfare monitoring system for the horse industry, the welfare of horses can be increased through improving awareness and stimulating changes in management.

The effect of long-term oral L-carnitine administration on insulin sensitivity, glucose disposal, plasma concentrations of leptin and acylcarnitines, and urinary acylcarnitine excretion in warmblood horses.

BACKGROUND: Insulin resistance in horses is an emerging field of interest as it is thought to be a contributing factor in the pathogenesis of many equine conditions. OBJECTIVES: The objectives of the present study were to determine the effects of long-term oral administration of L-carnitine on insulin sensitivity, glucose disposal, plasma leptin concentrations and acylcarnitine spectrum both in plasma and urine. ANIMALS AND METHODS: Six 3-year-old healthy warmblood geldings were used. In a double blind 2 × 2 Latin square design at a dosage of 100 mg/kg body weight (BW)/day for 28 days the effects of oral supplementation of L-carnitine (as fumarate) were assessed. Glucose disposal and insulin sensitivity were measured by means of the euglycemic-hyperinsulinemic clamp technique. Radioimmunoassays were used to determine plasma leptin and insulin concentrations. Electrospray tandem mass spectrometry was used to assess acylcarnitines both in plasma and urine. Statistical analysis was performed using a linear mixed-effects model and P values <0.05 were considered significant. RESULTS: Long-term L-carnitine administration did not affect insulin sensitivity. Plasma leptin and free carnitine concentrations in plasma and urine increased significantly (P = 0.047 and 0.000, respectively) following L-carnitine administration as well as short-chain acylcarnitines in plasma and urinary excretion of short- and medium-chain acylcarnitines. CONCLUSION AND CLINICAL RELEVANCE: Given the effects of oral administration of L-carnitine further clinical study is necessary in order to assess the potential beneficial effects in equine patients suffering from metabolic myopathies such as acquired multiple acyl-CoA dehydrogenase deficiency. IMPACT FOR HUMAN MEDICINE: The current study supports the treatment rationale of short-chain acyl-CoA dehydrogenase deficiency in humans with L-carnitine at an oral dosage of 100 mg/kg BW/day.

Amino acid profile during exercise and training in Standardbreds.

The objective of this study is to assess the influence of acute exercise, training and intensified training on the plasma amino acid profile. In a 32-week longitudinal study using 10 Standardbred horses, training was divided into four phases, including a phase of intensified training for five horses. At the end of each phase, a standardized exercise test, SET, was performed. Plasma amino acid concentrations before and after each SET were measured. Training significantly reduced mean plasma aspartic acid concentration, whereas exercise significantly increased the plasma concentrations of alanine, taurine, methionine, leucine, tyrosine and phenylalanine and reduced the plasma concentrations of glycine, ornithine, glutamine, citrulline and serine. Normally and intensified trained horses differed not significantly. It is concluded that amino acids should not be regarded as limiting training performance in Standardbreds except for aspartic acid which is the most likely candidate for supplementation.

The incidence and severity of gastric ulceration does not increase in overtrained Standardbred horses.

REASONS FOR PERFORMING STUDY: Gastric ulceration can be caused by different pathophysiological mechanisms including dietary factors, psychological stress and exercise. Overtraining is a medical syndrome in performance horses associated with altered hormone levels, altered feed intake, altered behaviour and decreased performance. These components might lead to a higher incidence of gastric ulceration in overtrained horses. OBJECTIVES: To investigate whether the incidence of gastric ulceration is increased in overtrained compared to control horses. METHODS: A longitudinal training study with twelve 1.5 years old Standardbred horses was performed on a treadmill for a total of 32 weeks. Training was divided into 4 periods: (1) acclimatisation (2) training (3) intensified training, and (4) detraining. In period 3, the horses were randomly divided into 2 groups: control (C) and intensified trained group (IT). At the end of each period, gastroscopy was performed in conscious horses after withholding feed for 12 h and water for 6 h using a 3.5 m video gastroendoscope. Lesion scores were assigned to areas of the stomach and graded 1-4. Logistic regression was used for statistical calculations. RESULTS: Evaluation of the stomach revealed only minor changes (grades 1 or 2) on each occasion. There were no significant differences in gastric lesion scores between groups or periods. Most lesions (70%) were found around the minor curvature. After detraining no lesions (0%) were found in contrast to periods 1 (40%, P = 0.056), 2 (30%) and 3 (30%). CONCLUSIONS: Experimentally-induced overtraining does not increase the incidence of gastric ulceration in normally fed Standardbred horses and detraining appears to reduce gastric ulceration.

The effect of long-term exercise on glucose metabolism and peripheral insulin sensitivity in standardbred horses.

REASONS FOR PERFORMING STUDY: To study the possible long-term effect of improved glucose tolerance in horses after long-term training, as the impact of exercise training on glucose metabolism is still unclear in the equine species. It is not known whether there is a direct long-term effect of training or if the measurable effect on glucose metabolism is the residual effect of the last exercise session. OBJECTIVES: To determine the chronic effect on glucose metabolism and peripheral insulin sensitivity of long-term training in horses by use of the euglycaemic hyperinsulinaemic clamp technique. METHODS: Eleven Standardbred horses were acclimatised to running on the high-speed treadmill for 4 weeks (Phase 1) followed by training for 18 weeks with an alternating endurance (approximately 60% HRmax) high intensity training programme (approximately 80% HRmax) (Phase 2). Training frequency was 4 days/week. At the end of Phase 1, a euglycaemic hyperinsulinaemic clamp was performed 72 h after the last bout of exercise in all horses. At the end of Phase 2, the horses were clamped 24 h or 72 h after the last bout of exercise. RESULTS: Glucose metabolism rate did not change significantly after 18 weeks of training, measured 72 h after the last exercise bout (0.018 +/- 0.009 and 0.022 +/- 0.006 mmol/kg bwt/min, respectively). Peripheral insulin sensitivity also did not change significantly following training (7.6 +/- 5.7 x 10(-6) and 8.0 +/- 3.1 x 10(-6), respectively). The same measurements 24 h after the last bout of exercise showed no significant differences. CONCLUSIONS: Results indicated that long-term training in Standardbreds neither changed glucose metabolism or insulin sensitivity 72 h after the last bout of exercise. POTENTIAL RELEVANCE: The fact that the beneficial effect of increased insulin sensitivity after acute exercise diminishes quickly in horses and no long-term effects on insulin sensitivity after chronic exercise have as yet been found in horses, implies that exercise should be performed on a regular basis in horses to retain the beneficial effect of improved insulin sensitivity.