Angiogenic markers in canine lymphoma tissues do not predict survival times in chemotherapy treated dogs.

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 266days, 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.

The size of the papilla of Vater and its relation to its height in comparison with the visibility of the sphincter of Oddi before and after cholecystectomy.

This study investigated the changes in the surface area and height of the papilla of Vater (PV) and the visibility of the sphincter of Oddi complex (SOK) using endoscopic ultrasound (EUS) examinations of 80 patients with symptomatic gallstones before and at 3 and 6 months after cholecystectomy. Following surgery, 50 patients experienced early atypical symptoms characteristic of postcholecystectomy syndrome (PCS) and 30 patients were asymptomatic. The PV measurements were greater in all patients compared with normal reference values and increased significantly from before surgery to 3 months postsurgery. At 6 months postsurgery, both the surface area and height of the PV had significantly decreased to preoperative values but remained higher than normal. The proportion of patients with a visible SOK had increased by 3 months postsurgery but had reduced again by 6 months. There were no significant differences between symptomatic and asymptomatic patients, suggesting that the changes observed in the PV and SOK do not explain the presence of the atypical symptoms of PCS.

Dynamic changes of echogenicity and the size of the papilla of Vater before and after cholecystectomy.

This study investigated the changes in echogenicity, as measured by endoscopic ultrasound, and the surface area of the papilla of Vater (PV) and their relationship with postoperative symptoms in a group of 80 patients with symptomatic gallstones before and at 3 and 6 months after cholecystectomy. After cholecystectomy, 50 patients experienced early atypical symptoms characteristic of postcholecystectomy syndrome (PCS) and 30 patients were asymptomatic. The surface area of the PV was larger than normal prior to surgery and increased after surgery. The healthy PV is isoechogenic, but 48% of all patients were anisoechogenic preoperatively, increasing to 61% at 3 months after surgery, and decreasing to 25% at 6 months postsurgery. There was no significant difference between the two patient groups, suggesting that the changes observed in the PV do not explain the presence of the atypical symptoms of PCS.

Non-cirrhotic and non-malignant acute extrahepatic portal vein thrombosis (PVT): short- and long-term results.

This observational cohort study reports the short- and long-term clinical outcomes of 31 patients admitted for acute non-malignant, non-cirrhotic portal vein thrombosis (PVT) over a 10-year period. Patients had a mean age of 43 years at admission and a mean duration of follow-up of 84 months. All patients were initially treated with anticoagulants. Complete recanalization occurred within 30 days after admission in 18 patients (58%), partially in nine patients (29%), and failed in four patients (13%). During follow-up, 10 patients (32%) had at least one episode of gastrointestinal bleeding. The probability of remaining bleed-free was 0.93 at 24 months and 0.61 at 48 months. Fundal varices were not controlled by endoscopic sclerotherapy, so all four patients underwent portosystemic shunt construction. To date, there has been no mortality. In conclusion, using a combination of different treatment options reduces the risk of death and late complications in patients with non-malignant, non-cirrhotic PVT.

The tyrosine kinase inhibitor sorafenib decreases cell number and induces apoptosis in a canine osteosarcoma cell line.

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.

The cranial instrument port in arthroscopy of the canine shoulder joint.

We 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.

Signalment differences in bone mineral content and bone mineral density in canine appendicular bones. A cadaveric study.

The 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. Mid-diaphyseal 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, body-weight of the male dogs was significantly higher compared to the females (P < 0.0001). Body weight and bone length were significantly associated with BMC and BMD (P < or = 0.023) in all bones but the radius. These data suggest that BMC and BMD appear to be highest in male large-breed dogs with a body weight greater than 30 kg. These results may help determine risk factors in fracture development and healing.

Not simply a foreign body.

The presence of foreign biological substances in the human body can lead to violent immune reactions. This is the report of a very rare case involving not only the presence of a biological substance, but also a symbiotic relationship between a living plant (the common wheat grain, Triticum aestivum L.) and the human body. Black coal particles and one cereal grain were removed from the subgalea of the right parietal region of a 35-year-old man who had sustained injuries in a motor vehicle accident 16 days earlier. There were signs of germination of the grain, but no macroscopic or microscopic evidence of an inflammatory reaction. Grain germination was verified microscopically. There are various explanations for the absence of an immune reaction, but only coal-tar-induced immunosuppression can explain the observed phenomenon.

Effects of radial shockwave therapy on the limb function of dogs with hip osteoarthritis.

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.

Expression of vascular endothelial growth factor and its receptors in canine lymphoma.

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 heterogeneous 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.

Schirmer tear test, phenol red thread tear test, eye blink frequency and corneal sensitivity in the guinea pig.

OBJECTIVE: To establish reference values for Schirmer tear tests (STT) I and II, phenol red thread (PRT) tear test and eye blink frequency, and to determine corneal sensitivity for normal guinea pigs. ANIMALS STUDIED: One hundred and eight eyes of 54 adult Duncan-Hartley guinea pigs. PROCEDURE: Schirmer tear test (STT) I and then STT II were performed in 36 guinea pigs. PRT and STT I were compared in 18 adult Duncan-Hartley guinea pigs. Corneal sensitivity was determined in 23 guinea pigs by evaluating the corneal touch threshold (CTT) of five different regions using a Cochet-Bonnet esthesiometer. Eye blink frequency was measured in 10 guinea pigs over a period of 20 min and in 17 guinea pigs over a period of 10 min. RESULTS: Mean STT I was 0.36 mm +/- 1.09 mm (wetting/min) and mean STT II was 0.43 mm +/- 1.29 mm (wetting/min). There was no significant difference between mean STT I and mean STT II (P = 0.79). The mean PRT-value was 16 +/- 4.7 mm (wetting/15 s), and the mean STT I-value in the same guinea pigs was 0.6 +/- 1.83 mm (wetting/min). Corneal sensitivity was significantly higher in the center than in the four limbal regions. The mean CTT for central, ventral, nasal, temporal and dorsal regions was 2, 1.7, 1.7, 1.7 and 1.6 cm or 3.7, 5.2, 5.6, 5.7 and 6.4 g/mm(2), respectively. Eye blink frequency was between two to five (mean 3.4 +/- 1.04) blinks per eye over 20 min in guinea pigs in their home environment, while in handheld and restrained guinea pigs eye blink frequency showed a variation between 0 and 17 blinks per eye (mean 3.24 +/- 3.64 blinks per eye) over 10 min. CONCLUSIONS: As there were no significant differences between STT I and STT II results, reflex tear secretion in the guinea pig may not exist. The most likely explanation is a lower corneal sensitivity in the guinea pig than in other species, such as cats, dogs and horses. Because of the small amount of tears, PRT is the preferred test for tear measurement in the guinea pig.

In vitro effects of meloxicam with or without doxorubicin on canine osteosarcoma cells.

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.

Voluntary and forced exercise influence the survival and body composition of ageing male rats differently.

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 (p<0.05). The S1 and TM groups gained most weight, the TM less after the age of 21 months (p<0.05). The body weights of the RW group was lower than those of the S1 and TM groups all the time (p<0.001) 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.

Housing conditions influence the survival and body composition of ageing rats.

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 6months 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 14months. H4 animals had significantly more fat from 11 to 20 months of age but lost weight, mostly fat when approaching 23months of age. The occiput-sacrum length of the animals increased up to 11months 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.

Ultrasonographic assessment of celiac disease in children: comparison with antiendomysium antibodies and histology.

UNLABELLED: Several attempts have been made to identify accurate screening tests for celiac disease (CD) with the aim to reduce the need for biopsy or obtaining appropriate selection criteria prior to intestinal biopsy. In this context we evaluated the usefulness of screening for endomysial antibodies (EMA) in the diagnosis of CD in children in comparison with CD-related changes detected by ultrasound (US) or biopsy. PATIENTS: We studied 35 children (1-15 years, 22 girls and 13 boys, mean age 8 years) with untreated CD (N = 15), treated CD (N = 10) and controls (N = 10), undergoing small bowel biopsy as a diagnostic procedure. METHODS: US of the small bowel was performed prior to mucosa biopsy using a 4- or 7-MHz transducer of a computed sonography device. The thickness of the intestinal wall and small bowel motility were recorded. Simultaneously, all children had serum routinely sampled for IgA EMA. RESULTS: All controls had histologically normal small bowel mucosa, US showed a small bowel wall thickness of 1 mm with normal motility in 9 children and non-specific wall changes in 1 child. EMA was weakly positive in 1 of these, and negative in all other controls. Of 15 children with untreated CD, severe enteropathy and strongly positive EMA were found in 10 cases; in 5 children moderate enteropathy and positive EMA were detected. Severe US changes were found in all of these children (1.6 mm thickness of the intestinal wall, hyperperistalsis and abundant fluid in the bowel). Mild enteropathy was found in 10 children with treated CD (3 months of a gluten-free diet). EMA was positive in 2 children and weakly positive in 8. Non-specific US changes were found in 6 children. In conclusion, our results indicate that US provides valuable information on small-bowel wall structure and can help in decision making on the necessity of small bowel biopsy. The present study confirms a stronger correlation between EMA and CD than between US and CD.

[Argon plasma coagulation (APC): a new mode in gastrointestinal endoscopy--first experience]. / Argonplasmakoagulation (APK): Neue Technik in der Endoskopie des Gastrointestinaltrakts--erste Erfahrungen.

Argon plasma coagulation (APC) is a new method of non-contact electrocoagulation in which current is applied to tissues by means of ionised argon gas (argon plasma). The development of special applicators has made this method applicable for gastrointestinal endoscopy. The primary indication for APC is the treatment of hemorrhage in the gastrointestinal tract. APC has been proven to be highly effective and easily used, with clear advantages over previously used methods. This article describes the introduction of APC in Slovenia and the first experiences with this method in the clinical department of internal medicine in Maribor.

Does long-term physical exercise counteract age-related Purkinje cell loss? A stereological study of rat cerebellum.

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 Cavalierìs 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.

The collagen biomarker of aging can be influenced by physical exercise also in senescent rats.

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 (100m/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 (400m/day from 24 to 27 and 200m/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.

Comparison between continuous and intermittent physical exercise on aging rats: changes in patterns of spontaneous activity and connective tissue stability.

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.