Increased expression of GAPDH protein is not indicative of nitrosative stress or apoptosis in liver of starved rainbow trout (Oncorhynchus mykiss)

Short-term starvation has been linked to in vivo protein degradation in liver of rainbow trout (Oncorhynchus mykiss). However, it is unclear whether this proposed increase in protein degradation is followed by programmed cell death (apoptosis) in liver of starved trout. A preliminary study in our laboratory revealed an isoform of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein that increased 4.5-fold in liver of starved trout. GAPDH is a glycolytic enzyme involved in other cellular functions, including apoptosis. Increased intracellular nitric oxide (NO) promotes nuclear translocation of GAPDH that is associated with increased apoptosis in mammals. If GAPDH protein is associated with apoptosis in rainbow trout, it could potentially be used as a biomarker of cellular stress in liver of teleost fish species. The purpose of this study was to determine whether increased GAPDH protein expression in liver of starved rainbow trout is associated with NO-induced apoptosis. Targeted proteomic analysis using multiple reaction monitoring (MRM) was used to determine the level of GAPDH in nuclear and cytoplasmic fractions and inducible nitric oxide synthase (iNOS) in cell lysates. Dot blot and DNA fragmentation analyses were conducted to evaluate protein S-nitrosylation and apoptosis, respectively. Results showed that cytoplasmic GAPDH was 3.4-fold higher in liver of starved versus fed rainbow trout but could not be detected in nuclear fractions. Starvation significantly reduced hepato-somatic index but had no effect on iNOS protein expression, protein S-nitrosylation, or apoptosis. Our results indicate that starvation promoted significant reduction in liver mass that was not associated with increased apoptosis or NO-induced stress and that greater GAPDH concentration in liver of starved rainbow trout was located primarily in the cytoplasm.     

Comparison of visual lameness scores to gait asymmetry in racing Thoroughbreds during trot in-hand

Regular monitoring of movement asymmetry with inertial measurement units (IMUs) to aid in the diagnosis of the underlying cause of a lameness is feasible. Normal ranges for specific groups of horses may be required, with consideration of expert veterinary opinions for both asymmetry screening and lameness assessment. The aim of this study was to determine movement asymmetry values compared with expert lameness scores to enable screening for lameness in Thoroughbreds in race training. IMU gait assessment during in-hand trot-up was performed in 25 racehorses undergoing routine gait analysis or lameness examination at the Singapore Turf Club. Video recordings were graded numerically (0–5) for lameness by six experienced racehorse veterinarians. Inter-observer agreement and consistency were determined. Median lameness scores were used to calculate sensitivity and specificity for head, withers and pelvic movement asymmetry. Guideline values for aligning movement asymmetry values with expert opinions about forelimb and hindlimb lameness were determined from receiver operating characteristics (ROC). Inter-observer agreement was poor to fair, inter-observer consistency was good (intraclass correlation coefficient: 0.667 for forelimbs and 0.617 for hindlimbs). ROCs indicated higher discriminative power for hindlimb lameness using pelvic asymmetry (90% sensitivity, 93% specificity) compared with forelimb lameness using head asymmetry (69% sensitivity, 89% specificity) or withers asymmetry (44% sensitivity, 89% specificity). When compared to expert lameness scores from videos of a limited number of Thoroughbred racehorses, preliminary guideline values for movement asymmetry screening for forelimb lameness (>|14.5 mm|) and hindlimb lameness (>|7.5 mm|) are higher than previously reported clinical thresholds of >|7 mm| for head movement and >|4 mm| for pelvic movement asymmetry.     

SMALL INTESTINAL EMPTYING TIME IN NORMAL BEAGLE DOGS

Gastric emptying time and small intestinal transit time in dogs are frequently discussed. However, it is often of interest to the radiologist to know what normal small intestinal emptying times should be. A total of 15 upper gastrointestinal studies was performed on five internal parasite-free, normal, standard Beagle dogs with three studies on each dog, 6 days apart. The ages and weights of the dogs ranged from 2–8 years and from 12.4–13.7 kg, respectively. Following 24-hour fasting, a dose of 10 ml/kg bw of 60% wt/vol barium sulfate suspension was administered through a stomach tube. Then, sequential radiographs were made at 30-minute intervals until the entire contrast medium column was in the colon and cecum. The mean, standard deviation, and range of gastric emptying time, small intestinal transit time, and small intestinal emptying time were 76 ± 16.7 (30–120), 73 ± 16.4 (30–120), and 214 ± 25.1 (180–300) minutes, respectively. This study offers the possibility that small intestinal emptying time may be used to further evaluate patients with suspected small intestinal partial obstruction, pseudo-obstruction, ischemia, or lymphangiectasia.     

Effects of a topical anaesthetic formulation and systemic carprofen, given singly or in combination, on the cortisol and behavioural responses of Merino lambs to castration

Objective   To determine the effectiveness of a topical anaesthetic formulation (Tri-Solfen) with or without the administration of a non-steroidal anti-inflammatory drug (carprofen) on the pain and distress response associated with ring or surgical castration of ram lambs.
Procedures   Merino ram lambs (n = 78) were allocated to 10 treatment groups: 4 groups of knife-castrated lambs and 4 groups of ring-castrated lambs received carprofen (4 mg/kg SC) and Tri-Solfen; 2 control groups (sham) received carprofen at 0 or 4 mg/kg SC. Measurements included plasma cortisol and haptoglobin concentrations, haematology, and behaviour, including posture.
Results   Knife-castrated lambs had higher peak cortisol and integrated cortisol responses for the first 6 h after treatment and greater concentration s of circulating acute phase proteins than ring-castrated lambs, both of which were significantly different from the sham controls. Tri-Solfen applied to the knife castration wound significantly reduced both the peak plasma cortisol concentration and the integrated cortisol response for the first 6 h and improved lying behaviour in the first 12 h. Carprofen reduced the cortisol response to knife castration at 30 min, but elevated the cortisol responses at 24 and 48 h. Carprofen nearly halved the number of acute pain behaviours associated with ring castration. There were no significant additive or synergistic effects from combining the analgesic treatments. Tri-Solfen applied to the tail wound provided no detectible benefits during ring castration + tail docking.
Conclusions   The physiological and behavioural responses suggest that ring castration has less impact on the lamb than knife castration. The specific analgesic treatments can provide modest amelioration of the pain and discomfort associated with castration. Alternative doses or application methods may enhance their efficacy.     

DUPLEX-ULTRASONOGRAPHIC EVALUATION OF THE COMMON CAROTID ARTERY IN THE RESTING, SEDATED AND ANESTHETIZED HORSE

The aim of this study was to determine the blood flow in the carotid artery during halothane anaesthesia by means of duplex-sonography. Seventeen warm blood horses were evaluated in dorsal and lateral recumbency and the results were compared with the values of the same horses resting and under sedation. The cross sectional area of the vessels, the time-averaged maximal blood velocity, time-averaged maximal blood flow and the resistance index were determined and the flow profile was evaluated. During halothane anesthesia the total blood flow shows a significant increase which is not dependent on the positioning of the horse. Mean blood velocity is decreased by sedation and significantly increased during anesthesia. Reduced peripheral resistance is expressed by a decline of the resistance index. After sedation and during recumbency there is an increase in diameter of the carotid artery.     

Plasma cortisol responses to three cortico-trophic preparations in normal dogs

Objective To compare cortisol responses to three corticotrophic preparations in normal dogs.
Animals Eight clinically normal dogs (four intact males, four intact females) of medium size.
Procedures Each dog received four treatments on four separate occasions in a duplicated Latin square pattern. Treatments were two adrenocorticotrophin (ACTH) preparations given intramuscularly at 2.2 U/kg, one of the ACTH preparations given intramuscularly at 1 U/kg and a synthetic polypeptide with ACTH-like activity (tetracosactrin, cosyntropin) given intravenously at 5 μg/kg. Plasma samples were taken for cortisol assay before and at 0.5, 1, 2 and 4 h after treatment.
Results Plasma cortisol concentrations were similar with the two ACTH preparations and at both dose rates. Tetracosactrin produced smaller mean peak cortisol concentrations, which tended to occur earlier than with ACTH, and smaller values for the area under the curve of plasma cortisol concentration from zero time to 4 h.
Conclusion The findings suggest that canine adrenal function can be tested adequately by giving ACTH intramuscularly at 1 U/kg and measuring plasma cortisol in samples taken at 0 and 2 h, or by giving tetracosactrin intravenously at 5 μg/kg and determining cortisol concentration at 0 and 1 h.     

Effects of delmadinone acetate on pituitary-adrenal function, glucose tolerance and growth hormone in male dogs

Objective To characterise the effects of delmadinone acetate on the pituitary-adrenal axis, glucose tolerance and growth hormone concentration in normal male dogs and dogs with benign prostatic hyperplasia.
Design A prospective study involving nine normal male dogs and seven with prostatic hyperplasia.
Procedure Delmadinone acetate was administered to six normal male dogs and seven dogs with benign prostatic hyperplasia at recommended dose rates (1.5 mg/kg subcuta-neously at 0, 1 and 4 weeks). Three normal controls received saline at the same intervals. Blood concentrations of ACTH, cortisol, glucose, insulin and growth hormone were measured over 50 days. Intravenous glucose tolerance and ACTH response tests were performed before and after treatment in the nine normal animals.
Results A substantial suppression of basal and 2 h post-ACTH plasma cortisol secretion was demonstrated after one dose in all dogs given delmadinone acetate. Individual responses after the second and third administration varied between recovery in adrenal responsiveness to continued suppression. Plasma ACTH concentration was also diminished after one treatment. No effects were evident on glucose tolerance or serum growth hormone concentrations.
Conclusion Delmadinone acetate causes adrenal suppression from inhibition of release of ACTH from the pituitary gland. Treated dogs may be at risk of developing signs of glucocorticoid insufficiency if subjected to stressful events during or after therapy. Neither glucose intolerance nor hyper-somatotropism seems likely in male dogs given delmadinone acetate at the recommended dose rate, but the potential for excessive growth hormone secretion in treated bitches remains undetermined.     

A longitudinal study of lymphocyte subsets in a cohort of cats naturally-infected with feline immunodeficiency virus

Despite the potential of feline immunodeficiency virus (FIV) as an animal model for human immunodeficiency virus (HIV) studies, the long term effects of naturally-occurring infection have not been determined. HIV infection causes an ongoing deterioration in immune function which directly correlates with disease, in particular acquired immunodefciency syndrome (AIDS). However, it is not known whether FIV-induced immunosuppression is progressive or related to the clinical condition. This study examined changes in lymphocyte subset numbers of serial samples, taken from cohorts of FIV-positive and FIV-negative cats over an 18-month period. FIV-positive cats were clinically staged as asymptomatic carriers (AC) or cats with AIDS-related complex (ARC), and FIV-negative cats matched and staged on the basis of similar diseases. During the course of the study, 4 FIV-positive cats developed AIDS, classed as the terminal stage of infection. There were no significant differences in the mean absolute numbers of any lymphocyte subset between the onset (to) and the completion (t18) of the study. Similarly there were no significant changes in subset numbers during the 18 months preceding the development of AIDS. While the study period was brief and the sample sizes small, it is postulated that FIV infection in Australia may not necessarily cause progressive immunodeficiency and that FIV-induced immunosuppression (as measured by subset analysis) may not be well correlated with the clinical status of the infected cat.     

Major compositional units of the moon: lunar prospector thermal and fast neutrons

Global maps of thermal and fast neutron fluxes from the moon suggest three end-member compositional units. A high thermal and low fast neutron flux unit correlates with the lunar highlands and is consistent with feldspathic rocks. The South Pole-Aitken basin and a strip that surrounds the nearside maria have intermediate thermal and fast neutron flux levels, consistent with more mafic rocks. There appears to be a smooth transition between the most mafic and feldspathic compositions, which correspond to low and high surface altitudes, respectively. The maria show low thermal and high fast neutron fluxes, consistent with basaltic rocks.