Laminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis

REASONS FOR STUDY: Xanthine oxidase (XO)-dependent production of superoxide anion and hydrogen peroxide, a characteristic of ischaemia-reperfusion injury, may contribute to the development of equine laminitis. OBJECTIVE: To determine the levels of XO and antioxidant enzymes (catalase, superoxide dismutase [SOD]) in the digital laminae of normal horses (CON) and horses in the developmental stage of laminitis using the black walnut extract (BWE) model. METHODS: Healthy horses (n = 12) were administered BWE (BWE group, n = 6), or water (CON group, n = 6) through a nasogastric tube. At the onset of leucopenia in the BWE-treated animals, all horses were anaesthetised, digital laminae and other samples collected rapidly and flash frozen, and the animals subjected to euthanasia. Extracts of the frozen tissues were assayed for the 2 conformational forms of xanthine: oxygen oxidoreductase (XOR), namely, xanthine dehydrogenase (XDH) and xanthine oxidase (XO), as well as the antioxidant enzymes, SOD and catalase. RESULTS: Extracts of liver, lungs and skin, but not digital laminae, from either CON or BWE-treated horses had endogenous SOD, whereas all had endogenous XO and catalase. The levels of XDH, XO and catalase were similar in extracts of laminae from CON and BWE-treated horses as was the ratio of XDH to XO in extracts. CONCLUSIONS AND POTENTIAL RELEVANCE: The absence of increased XO activity suggest against the involvement of this reactive oxygen intermediate-generating system in the development of laminar pathology in BWE-treated horses. Conversely, the absence of SOD from extracts of equine digital laminae, but not other tissues, suggests that the equine digital laminae are highly susceptible to damage by superoxide anion, produced, for example, by emigrant inflammatory leucocytes.     

Early laminar events involving endothelial activation in horses with black walnut- induced laminitis

OBJECTIVE: To determine proinflammatory gene expression, endothelial adhesion molecule gene expression, and matrix metalloproteinase (MMP) concentrations in laminar specimens at 1.5 hours after administration of black walnut extract (BWE) and to compare these values with later time points. ANIMALS: 25 horses. PROCEDURES: After nasogastric administration of BWE, anesthesia was induced at 1.5 hours in early time point (ETP) horses (n = 5), between 3 and 4 hours in developmental time point horses (5), and between 9 and 10 hours in acute onset of lameness time point horses (5). Anesthesia was induced at 3 and 10 hours after nasogastric administration of water in 2 groups of control horses (3-hour control group, n = 5; 10-hour control group, 5). Real-time quantitative PCR assay was performed on laminar specimens from control and ETP horses for cyclooxygenase (COX)-1, COX-2, interleukin (IL)-1beta, tumor necrosis factor-alpha, IL-6, IL-8, IL-10, MMP-2, and MMP-9 gene expression; and on laminar specimens from all groups for endothelial adhesion molecules, intercellular adhesion molecule (ICAM)-1, and E-selectin gene expression. Leukocyte emigration was assessed via CD13 immunohistochemistry, and gelatinase accumulation was determined by gelatin zymography. RESULTS: Laminar concentrations of IL-1beta, IL-6, IL-8, COX-2, ICAM-1, and E-selectin mRNA were significantly increased in ETP horses, compared with control horses. Concentrations of IL-1beta, IL-8, ICAM-1, and E-selectin mRNA peaked at 1.5 hours. In ETP horses, leukocyte emigration was present in 3 of 5 horses and pro-MMP-9 was detected in 2 of 5 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that endothelial activation and laminar inflammation are early events in laminitis; MMP accumulation likely is a downstream event.     

Estimating the mass of asteroid 253 mathilde from tracking data during the NEAR flyby

The terminal navigation of the Near Earth Asteroid Rendezvous (NEAR) spacecraft during its close flyby of asteroid 253 Mathilde involved coordinated efforts to determine the heliocentric orbits of the spacecraft and Mathilde and then to determine the relative trajectory of the spacecraft with respect to Mathilde. The gravitational perturbation of Mathilde on the passing spacecraft was apparent in the spacecraft tracking data. As a result of the accurate targeting achieved, these data could be used to determine Mathilde's mass as 1.033 (+/- 0.044) x 10(20) grams. Coupled with a volume estimate provided by the NEAR imaging team, this mass suggests a low bulk density for Mathilde of 1.3 grams per cubic centimeter.     

Blackspot bruise dependent changes in enzyme activity and gene expression in Lemhi russet potato

Blackspot bruise-induced changes in enzyme activity and gene expression were examined in the bruise susceptible cultivar Lemhi. The activities of several enzymes involved with the synthesis and metabolism of phenolic intermediates were examined over the time course of blackspot bruise development in tubers. A large, transient increase (200-fold) in phenylalanine ammonia lyase (PAL) activity was observed, while other enzymes examined (chorismate mutase, tyrosine ammonia-lyase, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase, and polyphenol oxidase) showed either modest or no activation. Maximal PAL activity was observed 48 hours after bruise induction, well after the discoloration reactions were complete. The change in PAL activity was associated with a transient increase in messenger RNA coding for PAL. In addition, messenger RNAs encoding two stress-induced genes, ubiquitin and the 70 kD heat shock protein, were transiendy induced by bruising. Physical impact also induced a marked decrease, followed by recovery, in the messenger RNA level for patatin, a primary storage protein in the tuber. These results suggest that the stress resulting in blackspot bruising elicits a wound response similar to those observed in other injuries to plant tissues.     

The confidential enquiry into perioperative equine fatalities (CEPEF): mortality results of Phases 1 and 2

Objectives To document the equine perioperative mortality rate and to highlight any factor associated with an increased risk of death up to 7 days after anaesthesia. Study design A prospective observational epidemiological multicentre study. Methods Data were recorded from all equidae undergoing general anaesthesia in 62 clinics. Power calculations indicated that 45 000 cases were required to detect the significance of important variables. Details of each horse, operation, anaesthetic agents and clinic personnel were recorded. Outcome at 7 days was recorded as: alive, put to sleep (PTS) or dead. Data were analysed by a standard multilevel logistic regression approach, considering the effects of clustering at the level of clinic. Results Data were collected from 41 824 cases over 6 years. A total of 39 025 (93.3%) were alive on day 7 and 785 were dead giving an overall death rate of 1.9% (95% CI: 1.8–2.0) and 2014 (4.8%) were PTS. About 5846 horses undergoing emergency abdominal surgeries (‘colics’) were excluded from subsequent analyses. A total of 35 107 ‘noncolic’ horses were alive at 7 days and 328 dead giving a death rate for noncolics of 0.9% (95% CI: 0.8–1.0). Five hundred and forty‐three (1.5%) noncolic horses classified PTS were excluded from further analyses. There were 109 (33%) deaths from cardiac arrest or post‐operative cardiovascular collapse, with 107 (32%) from fractures and myopathies. Fracture repair, out of hours surgery, and age below 1 month was associated with increased risk of dying whereas the use of acepromazine and intravenous anaesthetic agent maintenance of anaesthesia was associated with reduced risk. Conclusions A number of potential contributors to the high risk of anaesthetic‐related mortality have been identified. Further investigation of the underlying mechanism for their apparent harmful effects and development of alternative techniques is merited.     

Differentiation of gastrointestinal diseases in adult cattle.

Many of the common gastrointestinal disorders of adult cattle may be diagnosed by a careful physical examination, whereas other disturbances require the use of diagnostic testing. It is important to differentiate the causes of gastrointestinal disturbances to make better treatment decisions and have a clearer prognosis for the specific animal or herd of cattle.     

Distribution of insulin receptor and insulin‐like growth factor‐1 receptor in the digital laminae of mixed‐breed ponies: An immunohistochemical study

Reasons for performing study: Hyperinsulinaemia has been implicated in the pathogenesis of laminitis; however, laminar cell types responding to insulin remain poorly characterised. Objectives: To identify laminar cell types expressing insulin receptor (IRc) and/or insulin‐like growth factor‐1 receptor (IGF‐1R); and to evaluate the effect of dietary nonstructural carbohydrate (NSC) on their expression. Methods: Mixed‐breed ponies (n = 22) received a conditioning hay chop diet (NSC ～6%); following acclimation, ponies were stratified into lean (n = 11, body condition score [BCS]≤4) or obese (n = 11, BCS ≥7) groups and each group further stratified to remain on the low NSC diet (n = 5 each for obese and lean) or receive a high NSC diet (total diet ～42% NSC; n = 6 each for obese and lean) for 7 days. Laminar samples were collected at the end of the feeding protocol and stained immunohistochemically for IRc and IGF‐1R. The number of IRc(+) cells was quantified; distribution of IGF‐1R was qualitatively described. Laminar IRc content was assessed via immunoblotting. Results: The number of IRc(+) cells was greater in the laminae of high NSC ponies than low NSC ponies (P = 0.001); there was a positive correlation between the change in serum insulin concentration and number of IRc(+) cells (r²= 0.74; P<0.0001). No epithelial IRc(+) cells were observed; IRc(+) cells were absent from the deep dermis. Analysis of serial sections identified IRc(+) cells as endothelial cells. The distribution of IGF‐1R was more extensive than that of IRc, with signal in vascular elements, epithelial cells and fibroblasts. Conclusions: Increased dietary NSC results in increased laminar endothelial IRc expression. Laminar keratinocytes do not express IRc, suggesting that insulin signalling in laminar epithelial cells must be mediated through other receptors (such as IGF‐1R). Potential relevance: Manipulation of signalling downstream of IRc and IGF‐1R may aid in treatment and prevention of laminitis associated with hyperinsulinaemia.