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Objective—To assess natural variations in degree of refraction, corneal curvature, corneal astigmatism, corneal radius, and intraocular distance of healthy equine eyes. Animals—159 horses with healthy eyes that were admitted to a veterinary teaching hospital for nonophthalmic surgeries. Procedures—Eyes of horses were examined with a retinoscope prior to anesthesia and with a keratograph and A- and B-scan ultrasonographic biometers during surgery. In addition, manual caliper measurements of horizontal and vertical corneal radii were obtained. Results—Mean ± SD degree of refraction in the horizontal meridian of eyes was −0.06 ± 0.68 diopters (D). Vitreous body length and horse age correlated negatively with refraction values. The horizontal corneal radius (15.96 ± 1.28 mm) was larger than the vertical corneal radius (15.02 ± 1.09 mm). Accordingly, the vertical corneal curvature (21.56 ± 1.68 D) was greater than the horizontal corneal curvature (22.89 ± 1.65 D). Axial globe length (40.52 ± 2.67 mm), anterior chamber depth (6.35 ± 0.59 mm), lens thickness (12.30 ± 0.83 mm), and vitreous body length (21.87 ± 1.85 mm) were positively correlated with body weight, height, and age. Results of keratograph and caliper measurements correlated well for horizontal corneal diameter but poorly for vertical corneal diameter. Results of A- and B-scan ultrasonography differed by ≤ 1 mm in 64% of measured eyes. Conclusions and Clinical Relevance—Results of keratometry and ultrasonographic biometry varied widely. Additional research is needed to validate the keratograph used in our study for measurements in equine eyes.

Objective RANKL has been implicated in the pathogenesis of glucocorticoid-induced osteoporosis. This study was undertaken to evaluate the efficacy of denosumab, a neutralizing monoclonal antibody against human RANKL (hRANKL), in a murine model of glucocorticoid-induced osteoporosis. Methods Eight-month-old male homozygous hRANKL-knockin mice expressing a chimeric RANKL protein with a humanized exon 5 received 2.1 mg/kg of prednisolone or placebo daily over 4 weeks via subcutaneous slow-release pellets and were additionally treated with phosphate buffered saline or denosumab (10 mg/kg subcutaneously twice weekly). Two groups of wild-type mice were also treated with either prednisolone or vehicle. Results The 4-week prednisolone treatment induced loss of vertebral and femoral volumetric bone mineral density in the hRANKL-knockin mice. Glucocorticoid-induced bone loss was associated with suppressed vertebral bone formation and increased bone resorption, as evidenced by increases in the number of tartrate-resistant acid phosphatase (TRAP)–positive osteoclasts, TRAP-5b protein in bone extracts, serum levels of TRAP-5b, and urinary excretion of deoxypyridinoline. Denosumab prevented prednisolone-induced bone loss by a pronounced antiresorptive effect. Biomechanical compression tests of lumbar vertebrae revealed a detrimental effect of prednisolone on bone strength that was prevented by denosumab. Conclusion Our findings indicate that RANKL inhibition by denosumab prevents glucocorticoid-induced loss of bone mass and strength in hRANKL-knockin mice.

In this experiment, we investigated the long-term effects of a marginal zinc (Zn) supply on bone metabolism in aged rats. Nine-mo-old female Fischer-344 rats were divided into 8 weight-matched groups of 8 rats each. All rats were adapted for 1 mo to restrictive feeding (7.5 g/d) of a purified diet containing 8 g/kg sodium phytate and 64 mg/kg Zn. Control rats were pair-fed throughout the experiment. During the 1-mo depletion phase, controls received the Zn-replete diet with 64 mg/kg Zn, whereas Zn-deficient rats were fed the same diet with 2.2 mg/kg Zn. The depletion phase was followed by a 3-mo marginal phase in which the rats fed the diet with 2.2 mg/kg Zn received an additional daily Zn supplement of 75 microg Zn/rat by gavage. In the following 2-mo repletion phase, a marginal group was switched to the Zn-replete diet, while the other groups were maintained on marginal Zn supply or on the Zn-replete diet. Zn depletion and marginal Zn reduced serum and bone Zn and serum alkaline phosphatase activity. Zn repletion normalized serum Zn. However, apart from subtle changes in bone mineralization density distribution, Zn deficiency was not associated with detrimental effects on bone mineral density, turnover, architecture, or biomechanics relative to control rats at any time point. Our data suggest that Zn does not play an essential role in bone metabolism in aged rats and cast doubt on the hypthosis that Zn deficiency is a risk factor for osteoporosis.

SummaryWe describe the creation of a cranial instrument portal for shoulder joint arthroscopy in the dog using a target device that has been successfully employed for the caudal access. We compared the method with a conventional triangulation technique. The shoulder joints of 60 dog cadavers were investigated by arthroscopy and the two methods were compared on the basis of pre-defined parameters of outcome. Time, the number of trials, success, damage to cartilage and the accessibility of intra-articular structures were assessed. The anatomical localisation of the working cannula in muscles was also documented. In general, the new method permitted more successful insertion of the working cannula. This was achieved in a shorter period of time and was associated with fewer insertion attempts as well as less damage to joint cartilage. Intra-articular structures could be readily approached through the working cannula with both methods. Predetermined, clinically relevant points were comparatively more accessible with the new technique. Independent of the method, in only a few cases, the working cannula perforated neither muscles nor tendons when accessing the joint. It may be concluded that the new method using a target device is simpler and safer and offers a standardised technique to establish a cranial instrument port.

Microvessel density is a frequently used parameter of angiogenesis, which is a complex multistep process necessary for tumor progression. The aim of this study was to compare the microvessel density of normal lymph node biopsies with those diagnosed with lymphoma in dogs. Furthermore, we sought to determine if there was any correlation between microvessel density and vascular endothelial growth factor expression in canine lymphoma, representing a potential target for anti-angiogenic therapy. Combined immunohistochemistry (von Willebrand factor) and lectin histochemistry was used to highlight microvessels in 40 untreated canine lymphomas and 14 normal lymph nodes. To evaluate microvessel density, the number of profiles of blood vessels per unit area was calculated. Fifty image fields (a total area of 5.68 mm(2)) were sampled for each specimen in a systematic random, way. We found a significant difference between the microvessel densities (MVD) of normal and neoplastic lymph nodes (177+/-35 versus 241+/-72 microvessel profiles/mm(2)). Classifying lymphoma samples according to the working formulation and the Kiel classification system revealed no significant differences in MVD between different grade malignancies. Immunohistochemical demonstration of the proangiogenic protein vascular endothelial growth factor showed expression in 60% of canine lymphomas, although there was no correlation between microvessel density and vascular endothelial growth factor expression. As an increase in tumor angiogenesis was observed in lymphoma samples compared to normal canine lymph node tissue, additional anti-angiogenic therapy, besides conventional chemotherapy as a lymphoma treatment may be effective. The optimal target among many pro-angiogenic factors has yet to be elucidated.

SummaryThe objective was to determine signalment-related differences in bone mineral content (BMC) and bone mineral density (BMD) in dogs. Unilateral appendicular bones were harvested from 62 canine cadavers. Middiaphyseal regions of interest (ROIs) were scanned using a Hologic® DXA device Braincon, Vienna, Austria). BMC and BMD were calculated within this region. Middle-aged dogs (3 . 10 years) revealed the highest BMC and BMD levels. Mean BMC and BMD were higher in males compared to females. Furthermore, bodyweight of the male dogs was significantly higher compared to the females (P

Reactive oxygen species (ROS) play important roles in cellular senescence and organismic aging. Furthermore, they have been implicated in some of the adverse effects of chronic stress due to elevated peripheral levels of catecholamines. Here, we applied three different techniques to individually compare the systemic and intracellular oxidative stress in aged (23 months) and young (5 months) Sprague-Dawley rats, and in young rats treated for 12 or 24 h with norepinephrine (NE). Thiol groups of blood serum proteins (RSH) were determined by means of Ellman's reaction. Intracellular ROS were assessed in spleen cells and peripheral blood lymphocytes (PBL) by carbonylation of cellular (spleen) proteins as determined by immunoblotting (Oxyblot) and/or by means of 2',7'-dichlorofluorescein (DCF) fluorescence. As compared to the young, untreated controls, both old rats and NE treated young rats showed similarly lowered RSH values paralleled by elevated intracellular ROS levels or enhanced Oxyblot signals. Individual RSH values were highly significantly, negatively correlated with respective Oxyblot data as well as with DCF fluorescence. The results confirm the roles of ROS in aging and adrenergic stress in the rat model, and suggest that the decrease in RSH of blood serum may be taken as a valid indicator for the enhanced oxidative stress in lymphocytes.

Summary Vascular endothelial growth factor (VEGF) stimulates endothelial cell proliferation and has a pivotal role in tumour angiogenesis. The expression of VEGF and its receptors VEGFR-1 and VEGFR-2 was examined immunohistochemically in 43 specimens of canine lymphoma and in six normal lymph nodes. Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) were performed to detect VEGF protein and mRNA, respectively. VEGF protein was expressed by 60% of the tumours with diffuse cytoplasmic labelling of the neoplastic cells. Endothelial cells, macrophages and plasma cells were also immunolabelled. VEGFR-1 was expressed by variable numbers of neoplastic cells in 54% of lymphoma specimens. VEGFR-1 was also expressed by macrophages, plasma cells, reticulum cells, and vascular endothelial cells. Macrophages and lymphocytes in germinal centres of normal lymph nodes were also immunoreactive with anti-VEGF and VEGFR-1. Most tumours did not express VEGFR-2 but in 7% of sections there was focal labelling of neoplastic and endothelial cells, with a cytoplasmic and perinuclear pattern. The observed variability in expression of VEGF and its receptors probably relates to the fact that lymphoma is a heterogenous lymphoproliferative tumour. Individual differences in VEGF and VEGFR expression must be taken into account when VEGF and VEGFR-targeted approaches for anti-angiogenic therapy are considered in dogs.

The objective of this study was to evaluate the effects of extracorporeal radial shock wave therapy on the hindlimb function of dogs suffering from hip osteoarthritis. Twenty-four client-owned dogs with hip osteoarthritis were investigated; 18 of them received radial shockwave therapy and six were left untreated as controls. Force plate analysis on a treadmill was used to assess the dogs' hindlimb function before treatment and four weeks after the last treatment, and the treated dogs were re-evaluated three and six months after the treatment. The parameters chosen for evaluation were peak vertical force and vertical impulse, and the calculated symmetry indices. In the treated dogs, differences between the ground reaction forces exerted by the right and left hindlegs disappeared four weeks after the treatment, whereas in the control dogs only the peak vertical force distribution changed significantly. The significant improvement in the treated dogs was confirmed by changes in the symmetry indices. Significant improvements in vertical impulse and peak vertical force were observed three months after the treatment.

Background and aims: Moderate physical exercise and dietary restriction have both been demonstrated to delay some of the adverse effects of aging. In order to elucidate similarities or dissimilarities in their mode of action on the aging immune system in a comparative setting, we examined significant parameters of cell-mediated immunity in Sprague- Dawley rats. Methods: Male Sprague-Dawley rats, housed individually, were divided into four groups, living from 5 months (baseline group BL) up to 15, 19 and 23 months of age as follows: voluntary running in wheels (RW), food restricted by feeding to pair weight with RW animals (PW), forced running on treadmills (TM), and sedentary controls with ad libitum access to food (S1). White blood cell counts, capacity for lymphocyte proliferation in response to Concanavalin A, and interleukin-2 (IL-2) plasma concentrations were determined. Results: White blood cell counts and the cell numbers of lymphocytes, neutrophil and eosinophil granulocytes were significantly lower in the older RW and PW groups. We observed influences of forced exercise on lymphocyte proliferation: blastogenic reactivity was higher in TM animals compared with RW and PW animals at 23 months of age. Exclusively for RW animals, we found lower plasma concentrations of IL-2 at 23 months. Conclusions: Our data support the idea that moderate physical exercise modulates age-associated decline in the cell-mediated immunity of old Sprague-Dawley rats significantly more than corresponding dietary restrictions.

Cyclooxygenase (COX) inhibitors, already widely used to reduce fever, inflammation and pain, are under increasing consideration as potential agents for the prevention and treatment of neoplasia. As COX-2 was detected in human and canine osteosarcomas, we have evaluated the effect of the preferential COX-2 inhibitor meloxicam on an established D-17 canine osteosarcoma cell line, which expressed, as well as COX-1 and COX-2 also COX-3 (as demonstrated by Western blot). An XTT proliferation kit was used to assess surviving cells after drug treatment. At low concentrations (1, 2, 4 and 10 microm) meloxicam caused an increase in cell numbers while a marked anti-proliferative effect was observed at higher concentrations (100, 200 microm) after 3 days and also 3 weeks of incubation. The chemotherapeutic drug doxorubicin showed a cytotoxic effect at all concentrations (60-1920 nm). Exposure of tumour cells to combinations of meloxicam and doxorubicin revealed synergistic effects (with 240 nm doxorubicin), as well as sub-additive and antagonistic results, especially if combined with concentrations of meloxicam typically found in serum. Care should be taken in concluding, on the basis of one in vitro study, that meloxicam does not have a role in the treatment of canine osteosarcomas given that the results from in vivo studies may differ.

Background and Aims: Moderate physical exercise, dietary restriction as well as enriched environment in separate studies have been reported to delay some of the adverse effects of aging on brain function, parallel to an increase in brain-derived neurotrophic factor (BDNF). In order to elucidate these influences in a comparative setting, we examined the tissue concentrations of BDNF in the cerebral parietotemporal cortex of old Sprague-Dawley rats. Methods: Male Sprague-Dawley rats were divided randomly into six groups, living from 5 months (baseline group BL) up to 23 months of age as follows: voluntary running in wheels (RW), food restricted by feeding to pair weight with RW animals (PW), forced running on treadmills (TM), and sedentary controls with ad libitum access to food, either housed individually (S1) or in groups of 4 animals (S4). BDNF concentrations were determined by a commercially available ELISA. Results: We found higher BNDF concentrations in the 5 months old animals than in the 23 months old animals of group S1. The old sedentary group S4 showed significantly higher BNDF concentrations in comparison with the old individually caged groups RW, TM, PW and S1. Their BNDF concentrations were even higher than those of the young baseline group. Conclusions: Our data suggest that housing and social interactions have more influence on BDNF concentrations in the cerebral parietotemporal cortex of aging Sprague-Dawley rats than physical exercise and food restriction.

Background: Obesity combined with decreasing physical fitness in the ageing Western populations promotes a number of degenerative diseases, including chronic kidney disease. It has further been shown in rodent models that prevention of obesity by food restriction mitigates development of kidney lesions. Whether lifelong physical activity also has a positive effect is not known. Objective: To compare the effects of physical exercise and food restriction on the development of chronic kidney lesions in ageing rats. Methods: Male Sprague-Dawley rats were divided into groups: voluntarily running in wheels (RW), food restriction to the degree necessary to attain pair weight to RW rats (PW), forced running in treadmills (TM) and sedentary controls housed individually (S1) or 4 in each cage (S4). The interventions began at the age of 5 months and kidneys were sampled and analysed histologically at the ages of 15, 19 and 23 months. Results: Total score for kidney lesions (sum of the scores for glomerular changes, interstitial non-purulent inflammation, proteinaceous casts in tubules and increased amount of connective tissue, the possible maximum being 10.0) increased from 0.5 ± 0.2 at 5 months of age to 1.6 ± 0.3 for RW, 2.3 ± 0.4 for PW, 4.5 ± 0.4 for TM, 3.6 ± 0.5 for S1 and 5.4 ± 0.6 for S4 at the age of 23 months. The increase from 5 months of age was gradual for all groups through 15, 19 and 23 months. The patterns for the various lesions followed the same pattern with the exception of connective tissue, which did not increase. Conclusion: Voluntary running in wheels is as effective in mitigating kidney lesions as is food restriction, while forced running in a treadmill is not effective in this respect.

Background and aims: Obesity and lack of physical exercise characterize increasing numbers of people in Western societies, resulting in increasing morbidity and mortality. Regular physical exercise, on the other hand, has been shown to be beneficial. Beneficial effects have also been shown in laboratory rodent studies - lifelong physical exercise increases the mean life span but not the maximum life span, while food restriction increases both life spans. We studied male Sprague-Dawley rats in order to further analyze these beneficial effects. Methods: We compared voluntary running in wheels (RW) with food restriction to pair weight with RW (PW), and mild forced running in treadmill (TM) - all animals housed individually. Control groups were sedentary ad libitumfed animals housed individually (S1) and four to a cage (S4). The collagen biomarker of aging was calculated as a composite parameter from thermal stability data for 5 (baseline), 15, 19 and 23 months of age. Results: In RW and PW, the increase in the composite parameter was retarded to the same extent. The retardation in TM was less efficient. S4 had the highest value for the composite parameter, together with the highest body weight and fat content. In groupwise comparisons, the composite parameter tended to predict survival in the groups up to the age of 23 months. Conclusions: Voluntary running in wheels is more beneficial than mild-forced exercise on the treadmill. The composite parameter is a useful indicator of different interventions on aging rats.

The importance of maintaining physical fitness by engaging in exercise in a life-long perspective as well as the avoidance of obesity has been emphasised in recent years by epidemiological studies on human populations as well as studies on laboratory rodents. In laboratory studies, voluntary running in wheels and forced training in a treadmill have been used with beneficial results. Restriction of the food intake of sedentary laboratory rodents can be regarded either as life prolongation or prevention of life shortening by obesity. We compared the effects of these interventions on male Sprague-Dawley rats from the age of 5 to 23 months in the following groups: (1) RW=voluntary running in wheels; (2) PW=fed to pair weight with RW animals; (3) TM=forced training in a treadmill; and (4) S1=sedentary with ad libitum access to food. Each group consisted of 32 animals, all housed individually in cages. Two RW animals died, five died in each of the PW and S1 groups and 10 in the TM group (p0.05). The S1 and TM groups gained most weight, the TM less after the age of 21 months (p0.05). The body weights of the RW group was lower than those of the S1 and TM groups all the time (p0.001) and the difference increased all the time. Body composition was analysed by bioelectrical impedance analysis. There were no differences in fat free mass (FFM) neither between RW and PW at any time, nor between S1 and TM. FFM was lower for RW and PW compared to S1 and TM. TM gained FFM until the age of 17 months, while S1 gained FFM all the time. S1 gained fat all the time, but the gain for TM levelled off. It stayed constant for RW until 13 months and decreased afterwards. We conclude that voluntary running in wheels enhances survival and keeps body fat lower than in PW animals up to the age of 17 months. Body composition and survival data suggest that voluntary running is more optimal than forced. Care must, however, be taken in analyses, since RW is a heterogenous group because there is a large variation between the animals with respect to how much they run.

Ad libitum fed male Sprague-Dawley rats were randomly assigned to be housed individually (n=32) or four in each cage (n=32) from five to 23 months of age. Body weight and body composition by bioimpedance analysis were monitored longitudinally. An equation for analysis of body composition was developed. Deaths and causes of death were recorded. Of the individually housed animals (H1) 16 percent died, while 44 percent of those housed four in a cage (H4) died, the most common causes of death being circulatory failure, nephritis and tumours of the hypophysis. Body weight increased in both groups over time, significantly more for H4 from 6 months of age. Both fat free mass and total body fat increased in both groups over time. Fat in percent of body weight increased up to an age of 14 months. H4 animals had significantly more fat from 11 to 20 months of age but lost weight, mostly fat when approaching 23 months of age. The occiput-sacrum length of the animals increased up to 11 months of age and stayed then constant. Animals in both groups grew also in the cross-sectional dimension. The H1 animals continued to grow all the time, while the growth of the H4 animals stopped at the age of 17 months, when the mortality started to become pronounced. It is concluded that housing conditions can modulate the outcome of the ageing of sedentary and otherwise not treated rats in long-term experiments. Further, the feasibility of monitoring body composition longitudinally with bioimpedance analysis was demonstrated.

The normal processes of aging lead gradually to impairment of the function of most organs and to a gradually increasing force of mortality. This can to some extent be counteracted by life-long physical exercise as evidenced by epidemiological studies. The same is seen in animal experiments, where the mean survival time but not the maximum lifespan is increased by life-long physical exercise. In previous studies we have shown that the with aging increasing thermal stability of collagen—a biomarker of aging—is slowed down by life-long physical exercise in male Sprague–Dawley rats. Here we investigate whether late onset physical exercise also has a beneficial effect on this biomarker of aging. We found that mild exercise (100 m/day in a treadmill) retards the increase of thermal stability of collagen, when the rats are trained from 27 to 30 months of age. On the other hand more extensive exercise (400 m/day from 24 to 27 and 200 m/day from 27 to 30 months of age) has the opposite effect, which is in agreement with our previous study on rats trained from 12 to 24 months of age. We conclude that senescent rats can also benefit from physical exercise and that the optimum amount of training decreases with age.

Physical exercise affects properties of the central nervous system that may increase the brain's ability to counteract degenerative changes. We have previously reported that rats trained from 5 to 23 months of age have less age-related decrease in spontaneous motor activity than sham-treated sedentary rats. Each rat ran at a speed of 20 m/min on a horizontal treadmill, for 20 minutes, two times per day, 5 days a week. In the present study we have carried out stereological analyses of the cerebella of the same rats. The total number of Purkinje cells was estimated with the optical fractionator technique, the local volumes of individual Purkinje cells with the planar rotator technique, and the volumes of the cerebellar layers with Cavalieri`s principle. We found that sedentary aged rats have 11% fewer Purkinje cells and 9% smaller Purkinje cell soma volumes (both 2P = 0.02) than exercised aged rats, and that exercised aged rats have the same number of Purkinje cells as young rats. These findings indicate that the degree of age-associated degenerative changes in parts of the central nervous system is dependent on earlier life style and health habits and may be prevented or delayed by physical exercise. J. Comp. Neurol. 428:213–222, 2000. © 2000 Wiley-Liss, Inc.

We have previously shown that long-term regular physical exercise has a systemic influence on the rat by slowing the aging of its connective tissues, measured as thermal stability and biomechanical properties of tail tendons. This paper analyses whether the properties of limb muscle tendons are influenced not only by the aging process and the systemic effects of exercise but also from direct mechanical stimuli from long-term physical exercise. Male Sprague-Dawley rats were trained in a treadmill from the age of 5 to 23 months. The effects of training on muscle tendons were analyzed with respect to biomechanical properties. Also, the viscoelastic activation energies for interactions between collagen and the proteoglycan gel as well as between collagen fibrils were measured. Finally the asymptotes from the creep curves were calculated in order to estimate the magnitude of the viscoelastic creep. The effects of aging were analyzed with respect to the same parameters by comparing the group of 23-month-old sedentary rats with a 5-month-old baseline group. The biomechanical parameters did not change significantly with physical exercise. Neither did the activation energies change, but the asymptotes of the creep curves decreased, showing that there was less viscoelastic creep. Aging rendered the tendons significantly stronger and stiffer, increased the energy-absorbing capacity and decreased the strain values. The activation energies did not change with aging, but the high creep curve asymptote for the flexor tendons decreased. We conclude that aging rendered both types of tendons stiffer, and decreased their strain values at breaking point. Aging also increased the stress value, the energy absorption and the dry weight for the flexor tendon. Further, while physical exercise has a systemic delaying effect on age changes in connective tissues, in tendons subjected to substantial mechanical loads this effect as measured with biomechanical methods is counteracted by the optimization process elicited by the same physical exercise.

It has been shown that life-long, regular physical exercise has benefits for humans as well as laboratory animals. Population studies have shown that the longevity of humans is increased due to the decrease in all-cause mortality. Further, the functional capacities of organ systems, especially the cardiovascular, are maintained better. Body fat content stays lower. Similar conclusions have been drawn from animal studies, most of them using voluntary exercise in running wheels. We have previously shown that spontaneous activity, measured in an open field setting, is better preserved, suggesting a slowing of sensorimotor impairment with age, and possibly improved maintenance of dopaminergic and cholinergic systems. Further, the systemic effect on connective tissues was a slowing of the age-dependent increase of stability by changes in the cross-linking patterns. The purpose of the present investigation was to analyze whether late-onset training programs had any such effects, i.e., whether the aging effects seen in sedentary animals could be reversed to some extent. We trained male Sprague-Dawley rats from the age of 18 and 20 months until the age of 22 months, i.e., for 4 and 2 months, in a treadmill for 800 m/day. Spontaneous activity in an open field was assessed at the ages of 18, 20, and 22 months. For systemic changes in connective tissues, tail tendons were analyzed with respect to thermal stability and biomechanical strength parameters. The rats trained for 4 months lost weight significantly, which suggests that most of the fat accumulated during a sedentary life can be removed by physical exercise. Two months of training, either from the age of 18 or 20 months, had a positive effect on spontaneous activity, while the last 2 months of a 4-month training period had a negative influence. We conclude that the exercise program was too strenuous to be maintained for 4 months, which should be interpreted as a failure to increase the functional capacity sufficiently. Analysis of the thermal stability and biomechanical properties showed that both training programs moved these properties in a "younger" direction. We conclude that training starting late in life also influences the aging rat in a positive way, although there are limitations to the intensity of training that is beneficial.

Physical exercise has been shown to delay a number of aging changes and increase life expectancy but not maximum lifespan. We trained male Sprague-Dawley rats in a treadmill from the age of 5 months to 23 months. Up to this age the mortality (19%) was not different from that of the sedentary controls (16%). The body weights of the trained animals remained lower and reached a plateau, while those of the sedentary controls continued to increase. The spontaneous activity was assessed in an open-field setting every second month. Movement parameters (running distance, running speed, percentage of large and local movements), resting time and number of changes of direction were calculated. The running parameters were higher for the trained animals from the age of 12 months onwards, the relative difference increasing all the time. The resting time was lower for the trained animals, while the number of changes of direction did not change. It is suggested that this retardation of decline of spontaneous physical activity is due to a slowing of development of sensorimotor disturbances. These observations are compatible with a better preservation of cholinergic and dopaminergic activity.

To evaluate right intercostal Veress needle (VN) insertion for laparoscopy in dogs.Longitudinal cohort study.Female dogs (n = 56).The VN was inserted in the last palpable right intercostal space, either 1/3 (Group T; 28 dogs) or mid distance (Group H; 28 dogs) from the xiphoid cartilage to the most caudal extent of the costal arch. Problems encountered during VN insertion and injuries were recorded, graded, and compared between groups, and also between the first and last 20 insertions.Pneumoperitoneum was successfully achieved by VN insertion in 49 (88%) dogs after a single (45 dogs) or 2nd attempt (4 dogs). Frequency of complications was as follows: 20 grade 1 (subcutaneous emphysema, omentum, or falciform injuries); 6 grade 2 (liver or spleen injury), and 1 grade 3 complication (pneumothorax occurred). No significant difference was found between the 2 groups or between the first and last 20 dogs.Right intercostal VN insertion facilitates pneumoperitoneum in dogs with few consequential complications. No significant difference was found between entry sites; however, the mid distance insertion site in the last palpable intercostal space with dog positioned in dorsal recumbency is likely to result in less complications.

Objective To evaluate right intercostal Veress needle (VN) insertion for laparoscopy in dogs. Study Design Longitudinal cohort study. Animals Female dogs (n = 56). Methods The VN was inserted in the last palpable right intercostal space, either 1/3 (Group T; 28 dogs) or mid distance (Group H; 28 dogs) from the xiphoid cartilage to the most caudal extent of the costal arch. Problems encountered during VN insertion and injuries were recorded, graded, and compared between groups, and also between the first and last 20 insertions. Results Pneumoperitoneum was successfully achieved by VN insertion in 49 (88%) dogs after a single (45 dogs) or 2nd attempt (4 dogs). Frequency of complications was as follows: 20 grade 1 (subcutaneous emphysema, omentum, or falciform injuries); 6 grade 2 (liver or spleen injury), and 1 grade 3 complication (pneumothorax occurred). No significant difference was found between the 2 groups or between the first and last 20 dogs. Conclusions Right intercostal VN insertion facilitates pneumoperitoneum in dogs with few consequential complications. No significant difference was found between entry sites; however, the mid distance insertion site in the last palpable intercostal space with dog positioned in dorsal recumbency is likely to result in less complications.

Background: The pathophysiology of age-related bone loss and whether age-related bone loss can be prevented by exercise are still a matter of debate. Objective: It was the aim of this study to investigate the long-term effects of exercise and mild food restriction on bone mineral density (BMD) and bone geometry in the appendicular skeleton of aging male rats. Methods: Male Sprague-Dawley rats were studied from 5 to 23 months of age. The rats were divided into 4 groups: baseline, free access to food and running wheels (RW), fed to pair weight with the RW group (PW) and sedentary control animals with free access to food (SED). All rats were housed individually. Volumetric BMD and geometry of femurs and tibiae were assessed by peripheral quantitative computed tomography (pQCT). In addition, the tibial shafts were analyzed by cortical bone histomorphometry. Results: At the end of the experiment, RW and PW rats had similar body weight. The body weight of SED rats was 31% greater than that of RW rats. pQCT analysis of femurs and tibiae as well as histomorphometric analysis of the tibial shafts showed that dietary restriction resulted in an enlargement of the marrow cavity and cortical thinning at the femoral and tibial shafts relative to the RW and SED groups. Voluntary running exercise provided no additional protection against age-related bone loss when compared with the 31% heavier SED control rats. Neither exercise nor increased body weight in SED animals could completely prevent age-related bone loss between 19 and 23 months of age. Conclusion: We conclude that dietary restriction had clear negative effects on BMD and bone geometry and that running wheel exercise provided partial protection but could not prevent age-related bone loss.

Angiogenesis, which is essential for malignancies to progress, depends on various signalling proteins including vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptors 1 and 2 (VEGFR-1 and VEGFR-2). Microvessel density (MVD) is frequently used to evaluate angiogenesis. This study assessed the relationship between expression of VEGF, VEGFR-1 and VEGFR-2, MVD and the survival time in dogs with lymphoma. VEGF, VEGFR-1 and VEGFR-2 expression was evaluated immunohistochemically and microvessel profiles were counted in 34 lymphoma samples. Seventy-nine percent of the samples showed high VEGF expression and 62% were highly positive for VEGFR-1; VEGFR-2 immunoreactivity was mostly negative. Dogs treated with chemotherapy had a median survival time of 266 days, but no significant relationships were found between overall survival time, MVD and expression of VEGF, VEGFR-1 or VEGFR-2. In this study, VEGF its receptors and the MVD were no prognostic factors in dogs with lymphoma.

FINDIK, Murat/0000-0003-1408-2548 WOS: 000270183000005 PubMed: 19781024 Objective To compare surgical times and perioperative complication rates of single portal access and 2-portal laparoscopic ovariectomy (LapOVE) in dogs using a bipolar vessel sealer/divider device, and to evaluate the performance of novice laparoscopists for right ovariectomy. Study Design Controlled clinical trial. Animals Female dogs (n=42). Methods Dogs were divided into groups: 1=single portal and 2=2 portal. LapOVE was performed using a 5 mm vessel sealer/divider device and a 10 mm operating laparoscope (Group 1) or a 5 mm laparoscope (Group 2). Dog characteristics (weight, body condition score, ovarian ligament fat score), operative time, and perioperative complication rate were compared between groups. Right ovariectomy duration was evaluated for 2 novice laparoscopists. Results No significant difference was found in mean total surgical time between group 1 (21.07 min/s) and group 2 (19.06 min/s). Factors significantly affecting times included body condition scores, ovarian ligament fat score, ovarian bleeding, and surgeon expertize. Minor complications (bleeding from ovaries or after splenic trauma) occurred and were similar in both groups. Bleeding was correlated to body condition score and ovarian ligament fat score. Interindividual differences were found among surgeons for right ovariectomy time. Conclusions Single portal access LapOVE using vessel sealer/divider device is feasible, safe, and does not significantly increase total surgical time in comparison with 2-portal approach. Laparoscopic skills may play a role in ability to perform single portal LapOVE. Clinical Relevance LapOVE can be performed using single portal access. Department of Obstetrics and Gynaecology, Faculty of Veterinary Medicine, Ondokuz Mayys UniversityOndokuz Mayis University [55139] This project was realized at the Department of Obstetrics and Gynaecology, Faculty of Veterinary Medicine, Ondokuz Mayys University, 55139 Samsun, Turkey.

Objective—To compare values for tibial plateau angle (TPA) obtained in dogs by conventional and digital methods. Design—Evaluation study. Animals—37 dogs with stifle joint abnormalities. Procedures—In all dogs, radiographs of both stifle joints were obtained by conventional and digital means. On conventionally acquired radiographs, TPA was measured with a protractor and fine-point pencil in accordance with standard guidelines. A software program was used to measure TPA on digitally acquired radiographs. Two viewers with different levels of experience performed all measurements 3 times. Results—For both viewers and both limbs, conventional TPA measurements were significantly correlated with digital measurements all 3 times. Conventional and digital measurements obtained by viewer 1 were significantly different from values obtained by viewer 2. However, inter-viewer and intertechnique differences in TPA measurements resulted in rotational differences of < 1 mm. Conclusions and Clinical Relevance—Results suggested that TPA measurements obtained with the digital method were comparable to those obtained by use of the conventional method. Subjectively, the digital method was easier to perform and faster and produced better-quality images.

Canine osteosarcoma, an aggressive cancer with early distant metastasis, shows still despite good chemotherapy protocols poor long term survival. The aim of our study was to determine whether sorafenib, a novel multikinase inhibitor, has any effect on D-17 canine osteosarcoma cells. A cell proliferation kit was used for detecting surviving cells after treatment for 72 h with sorafenib or carboplatin or their combination. A significant decrease of neoplastic cells was observed after incubation with 0.5-16 microM sorafenib or with 80-640 microM carboplatin. Using immunocytochemistry for activated caspase 3 to evaluate apoptosis, we found significantly more positive cells in the sorafenib treated groups. Paradoxically, expression of the nuclear proliferation marker Ki-67 was also significantly higher in sorafenib treated cells. The drug sorafenib showed potent antitumour activity against D-17 canine osteosarcoma cells in vitro, suggesting a potential as a therapeutic tool in the treatment of bone cancer in dogs.

Udgivelsesdato: 2008-Jul Systematic protein expression studies in the brain of exercising and sedentary animals have not been carried out so far and it was therefore decided to determine differences in metabolic protein levels in rat hippocampus of sedentary, voluntary and involuntary exercising rats by a proteomic approach. Aged, male Sprague-Dawley rats, 23 months old, were used for the study: the first group consisted of sedentary rats, the second of rats with voluntary exercise from five to 23 months and the third group was performing involuntary exercise on a treadmill from five to 23 months. Two-dimensional gel electrophoresis with subsequent mass spectrometrical identification of spots followed by quantification of spots was carried out. Identification of significantly differential proteins was validated by the determination of the corresponding enzyme activity. Five individual metabolic proteins showed differential protein levels in the three groups: mitochondrial precursors of ornithine aminotransferase, isocitrate dehydrogenase [NAD] subunit alpha, malate dehydrogenase, ubiquinol-cytochrome-c reductase complex core protein 1, and ubiquitin carboxyl-terminal hydrolase isozyme L1. The unambiguously identified metabolic proteins were mainly of mitochondrial localization and fit the expectations of altered mitochondrial activity in exercise. Reduced ubiquitin carboxyl-terminal hydrolase isoenzyme L1 levels in treadmill (forced) exercise show the involvement of the proteasomal pathway as a novel finding. These results not only form the basis for functional studies elucidating mechanisms and differences between voluntary and forced exercise in hippocampal metabolism but also highlight the most intriguing aspect that exercise is affecting the brain at the protein level.

Udgivelsesdato: 2008-Mar Information about protein expression studies in the brain of exercising and sedentary animals is limited. Cognitive functions change during exercise and the aim of this study was to investigate rat protein levels of the protein machinery in the hippocampus, the main cognitive brain area for spatial learning and memory, in exercising rats. Protein fluctuations may reflect functional variation during exercise. Male Sprague-Dawley rats, 23 months old, were used for the study: the first group consisted of sedentary rats, the second of rats undertaking voluntary exercise from 5 months to 23 months and the third undertaking involuntary exercise on a treadmill from 5 months to 23 months. Two-dimensional gel electrophoresis with subsequent mass spectrometrical identification assigning spots to proteins and determination of coomassie-densities was carried out. Heterogeneous nuclear ribonucleoprotein K, one protein variant of heat shock cognate 71 kDa protein and BAG family molecular chaperone regulator 5 showed differential protein levels in the three groups when a p-value of

Udgivelsesdato: 2008-Jun Animal models for male osteoporosis are scarce. This study aimed at identifying the impact of different living conditions on bone structure and metabolism as well as the inflammatory status in a rat model of age-related male osteoporosis. Bone mineral density, bone histomorphometric data, ex vivo osteoclast generation, and bone metabolism serum marker as well as intracellular cytokine expressions were evaluated in 23-month-old male Sprague-Dawley rats subjected to different housing conditions from the age of 5 months. Running rats were housed individually and were exercised voluntarily in running wheels attached to their cages. Dieting rats were housed individually, too, but were fed to pair weight with the running rats. Walking rats were exercised mildly by use of a treadmill (800m/day, 5 days a week) and social rats were kept as four in a cage and fed ad libitum. Whereas no marked differences could be found for bone mineral density, trabecular bone volume as well as trabecular bone surface were diminished in walking rats. The ex vivo osteoclast generation assay revealed no significant differences between groups. Osteoblasts of running rats were not only decreased in number, but displayed also a lower activity as indicated by decreased serum osteocalcin levels. Osteoclast activity was increased in the same group as indicated by elevated CTX (c-terminal telopeptide of type I collagen) levels. Additionally, production of tumor necrosis factor (TNF)-alpha and interferone (IFN)-gamma by CD8(+) T cells was elevated in running rats. In conclusion, running has a negative effect on bone metabolism and proinflammatory status in male aged rats.

Background: Voluntary running in wheels as well as food reduction increase the life spans of rats. Disparate parameters such as the collagen biomarker of ageing and the development of kidney pathologies are decreased by voluntary exercise. There are few reports on the influence of physical exercise and food restriction on the skeleton of male rats. Most investigations initiated rather short-term interventions in 4- to 5-week-old animals and thus studied more the influence of growth than the influence of ageing on the skeleton. Objective: To compare the effects of physical exercise and food restriction on the biomechanical properties of bone tissue of ageing male rats with the interventions starting at the age of 5 months with the end point at 23 months. This enables the study of the influence of these interventions on the ageing of the skeleton. Methods: Five groups of male Sprague-Dawley rats were used: baseline (BL), voluntarily running in wheels (RW), food restriction to attain pair weight with RW animals (PW), forced running in treadmills (TM), and sedentary controls (SE). The biomechanical properties of femoral neck, diaphysis, and distal metaphysis were measured. Results: While the body weights and fat-free mass increased from BL to SE group, the occiput-sacrum length did not increase and the length of the femur increased marginally. These lengths were slightly retarded in RW and PW groups compared to the SE group. The strength of the distal femoral metaphysis decreased from BL to SE group. This decrease was counteracted by physical exercise (RW and TM groups) as well as by food restriction (PW group). In contrast, the strength of the femoral mid-diaphysis did not differ between BL and SE groups. Conclusions: The distal metaphysis in the male rat femur is more prone to decreasing biomechanical strength than the diaphysis during ageing. Physical exercise, when started at the age of 5 months, when the skeleton has reached its adult size, is somewhat effective in counteracting these changes. There is also some retarding effect of food restriction.

Osteoporosis is a common and severe condition in elderly men, which is poorly characterized. In order to identify the hallmarks of age-related bone loss in the male mammalian skeleton, we studied several aspects of bone structure and metabolism in 23-month-old male Sprague–Dawley rats and compared them to 5-month-old animals. Cancellous bone mineral density, bone volume and trabecular number were markedly reduced in the proximal tibia of aged rats when compared to the young rats. An increase in bone matrix material density indicating a reduced deposition of new bone matrix was seen. Also, serum levels of osteocalcin, a marker of bone formation, were reduced in old males. The decreased bone formation could in part be linked to the decreased serum insulin-like growth factor 1 (IGF-1) levels which were observed in these animals. Serum levels of RatLaps™ (c-terminal telopeptide of type I collagen) were increased. Interestingly, an ex vivo osteoclast generation assay revealed that bone marrow from aged rats formed fewer osteoclasts than that from young rats. Consistent with this observation, serum levels of soluble RANKL, a critical osteoblast derived factor for osteoclastogenesis, were decreased in aged rats and RANKL mRNA expression was slightly reduced in bone marrow cells. Elevated leptin and adiponectin levels present in these animals could have contributed further to impaired osteoclastogenesis. We conclude that aged male rodents are characterized by a severely diminished cancellous bone network and a bone turnover situation in which bone formation is decreased to such an extent that it is outweighed by bone resorption, despite a blunted osteoclast generation potential of the bone marrow.

Systematic protein expression studies in the brain of exercising and sedentary animals have not been carried out for far. Signaling proteins are main structures regulating hippocampal function and we decided to determine differences in signaling protein levels in rat hippocampus by a proteomic approach. Aged, male Sprague-Dawley rats, 23 months old, were used for the study: the first group consisted of sedentary rats, the second of rats with voluntary exercise from 5 to 23 months and the third was performing involuntary exercise on a treadmill from 5 to 23 months. 2-DE with subsequent mass spectrometrical identification of spots followed by quantification of spots was carried out. Annexin A5, A3, phosphatidylethanolamine-binding protein, guanine nucleotide-binding protein G(I)/G(S)/G(T), 14-3-3 protein gamma, 14-3-3 protein zeta/delta, prohibitin, visinin-like 1, protein phosphatase 1, septin 8, phosphoprotein enriched in astrocytes 15, transcription factor Pur-beta, EEA1 protein, SH3 domain-binding glutamic acid-rich-like protein 2, and cell division cycle 42 showed differential protein levels in the three groups. These results form the basis for functional studies elucidating mechanisms and links between exercise and hippocampal signaling and function.

Spatial learning and memory decline with ageing in humans as well as rats. We examined the influence of different interventions on male Sprague Dawley rats with respect to learning ability and memory at the age of 5, 10 and 18 months. The intervention and control groups were: (RW) voluntary exercise in running wheels, (PW) sedentary, food restricted (by about 25%) to keep them at pair weight with RW, (S1) sedentary, fed ad libitum, (TM) forced training in a treadmill, and, (S4) sedentary, fed ad libitum. The animals in all groups were housed individually except those in group S4, which were housed four in each cage. The ability of learning and memory was determined in the Morris water maze. The results showed a significantly better learning ability when young in comparison with their ability when having grown older. At the age of 18 months, the performance was significantly better in the subgroups which had received training also at the age of 10 months compared to the subgroups receiving their first training. None of the various interventions had any significant effect on these functions. Repeated training seems to be the best intervention with respect to retaining learning ability and memory.

Succinic dehydrogenase (SDH) activity, preferentially evidenced by cytochemical methods, has been measured by computer-assisted morphometry in the heart muscle of old sedentary and age-matched animals chronically undergone physical exercise (20 min, twice a day, 5 days a week). The area of the SDH-positive mitochondria (MA) and the overall area of the cytochemical precipitates due to SDH activity (PA) were semiautomatically measured and the ratios PA/MA as well as MA/overall myocardial tissue area analysed (MA/TA) were the parameters taken into account. No significant difference was found between the two groups investigated as regards PA/MA, whereas the MA/TA value is significantly increased in the animals undergone physical training. The present findings document that chronic physical exercise significantly increases the overall mitochondrial area involved in energy provision in the old myocardial tissue. Considering that myocardial function highly relies on mitochondrial metabolism, our results support a beneficial effect of chronic physical exercise on the old heart muscle.

Regular physical exercise has emerged, together with dietary restriction, as an effective intervention in delaying degenerative diseases and augmenting life span in rodents. The mechanisms involved remain largely unknown, although a beneficial influence on the age-related alteration of insulin sensitivity has been hypothesized. As muscle triglyceride (TG) accumulation is considered a reliable index of muscle insulin resistance, in this study we explored muscle TG content in 23-month-old male Sprague-Dawley rats subjected to life-long training. Plasma glucose, insulin, free fatty acid (FFA) and leptin levels were also measured. Both voluntary running in wheels (RW) and forced training in treadmill (TM) were studied. As RW rats weighed less than controls, a cohort of untrained animals, fed to pair weight (PW) with RW, was added to discriminate the effect of exercise from that of food restriction. Sedentary ad libitum fed rats served as controls. In 23-month-old RW rats, muscle TG content was reduced by 50% with respect to age-matched sedentary controls, while in TM group this reduction was smaller but still highly significant, and occurred independently on the changes in body fat mass. In both the trained rat groups, there was a significant decrease in circulating FFA levels and a trend to reduced insulin levels. In PW rats, muscle TG levels decreased similarly to RW rats, while plasma parameters were less modified. In particular, RW training was more effective than PW in preventing the age-related increase in circulating leptin levels. Our results suggest that voluntary exercise effectively counteracts the development of insulin resistance in the muscles of ageing rats as well as other related changes such as hyperlipacidaemia and compensatory hyperleptinaemia. Forced training or moderate food restriction appear slightly less effective than voluntary exercise in preventing age-dependent alterations in nutrient distribution and/or utilization.

Life-long physical exercise increases the mean but not maximum life span of laboratory rodents. It further improves the performance in an open field setting, which suggests a delay of sensorimotor disturbances, and retards the aging increase of thermal stability of collagen. The purpose of the present study was to examine whether the amount or continuity of training is more important. We trained male Sprague-Dawley rats for 12 months from the age of 12 months in a treadmill: 1) continuously for 3 km/week (total distance covered: 144 km); 2) intermittently for 3 km/week for 8-week periods with 8-week resting periods in between (72 km), and 3) intermittently for 6 km/week for 8-week periods with 8-week resting periods in between (144 km). All training regimens prevented the increase in body weight seen in sedentary animals, although the intermittent training programs caused fluctuations with the training and resting periods. With respect to spontaneous movements in an open field setting, continuous training was most effective in counteracting age-related changes. Continuous training was also most effective for delaying age-related increase in thermal stability of collagen. We conclude that the continuity of training is more important than the amount and intensity of training.

Regular physical exercise has been shown to have a number of benefits compared with sedentary behaviour, such as delaying a number of aging changes and increasing the life expectancy but not the maximum lifespan. The purpose of this paper is to analyze the effects of physical exercise on the connective tissues of the body, especially systemic effects. We trained male Sprague-Dawley rats in a treadmill from the age of 5 months to 23 months. We analyzed the effects of training on tail tendons with respect to thermal stability of collagen and biomechanical properties of tendon bundles. Although tail tendons are attached to muscles, the are not weight-bearing as limb muscle tendons and can, therefore, be considered to be subjected mainly to systemic effects. The thermal stability of tail tendon collagen was significantly lower or ‘younger’ for the trained group compared with the sedentary one. The biomechanical parameters were likewise ‘younger’ with respect to maximum stress, although considerably stiffer compared with a 5-month-old base-line group. This suggests that there are several mechanisms, elicited by physical excercise, that act on the connective tissues. It can be concluded that life-long physical exercise has a beneficial influence on the connective tissues of the maturing and aging organism.