Evaluation of the induction of vasoactive mediators from equine digital vein endothelial cells by endotoxin

OBJECTIVE: To determine the effect of endotoxin (lipopolysaccharide [LPS]) on vasoactive mediator production by cultured equine digital vein endothelial cells (EDVECs). SAMPLE POPULATION: EDVECs obtained from forelimb digital veins of 7 healthy adult horses. PROCEDURES: EDVECs were incubated with or without LPS (1 microg/mL) for 0, 2, 4, 6, 22, and 24 hours. The EDVECs were incubated for 18 hours with LPS (10 pg/mL to 1 microg/mL) with or without ibuprofen, cycloheximide, or L-nitroarginine methyl ester. Medium concentrations of prostacyclin, cyclic guanosine monophosphate, endothelin-1, and thromboxane A(2) were determined. Changes in inducible nitric oxide synthase and cyclooxygenase-2 expression were determined. RESULTS: LPS stimulated mean 4.2- and 14.1-fold increases in EDVEC prostacyclin and cyclic guanosine monophosphate production, respectively, after 22 hours. These effects were LPS concentration-dependent (LPS concentrations that induced a response halfway between the maximum response and baseline of 1.50 and 1.22 ng/mL, respectively). The LPS-induced cyclic guanosine monophosphate production was significantly inhibited (to basal concentrations) by L-nitroarginine methyl ester, and prostacyclin production was inhibited by cycloheximide and ibuprofen. Production of thromboxane A(2) by EDVECs was not detected. Endothelin-1 accumulated in the medium, but LPS did not enhance its production. Inducible nitric oxide synthase expression in EDVECs was not detected with the available antibodies, whereas LPS stimulated cyclooxygenase-2 expression in a time- and concentration-dependent manner. CONCLUSIONS AND CLINICAL RELEVANCE: LPS stimulated vasoactive mediator production by equine endothelial cells, which may play a role in LPS-induced digital hypoperfusion.     

Corneal epithelial inclusion cyst in a Llama

A 13-year-old, female Llama presented for evaluation of a limbal based corneal mass involving the OD of 4 months duration. The mass was excised en bloc by a nonpenetrating keratectomy, followed by placement of a conjunctival advancement flap covering the keratectomy site. The mass was submitted for histological evaluation. Histopathology identified the mass to be a corneal epithelial inclusion cyst filled with necrotic squamous and neutrophilic debris. Surgical excision was complete and considered curative with no signs of recurrence 3 months postoperatively. There was no known prior ocular trauma; however, a previously performed corneal biopsy for evaluation of recurrent epithelial erosions may have been an initiating cause.     

Effects of agricultural management on nematode–mite assemblages: Soil food web indices as predictors of mite community composition

Biological indicators based on abundances of soil organisms are powerful tools for inferring functional and diversity changes in soils affected by agricultural perturbations. Field plots, combining organic and conventional practices with no tillage, conservation tillage and standard tillage maintained different nematode assemblages and soil food webs. Soil food web indices based on nematode assemblages were reliable predictors of the trophic composition of functional characteristics of soil mite assemblages. Bacterial-feeding and predatory nematodes, together with predatory mites, were abundant in the organic-no till treatments and were associated with high values of the Enrichment and the Structure Index based on nematode assemblages. Conventional-Standard tillage treatments had high abundances of fungal- and plant-feeding nematodes and algivorous mites, associated with high values of the Basal and Channel Index. This study validates the hypothesis that nematode-based soil food web indices are useful indicators of other soil organisms such as mites, with similar functional roles and environmental sensitivities.     

Metal chelate molluscicides: The redistribution of iron diazaalkanolates from the gut lumen of the slug,Deroceras reticulatum (Müller) (Pulmonata: Limacidae)

Two related iron chelates, one toxic to slugs by ingestion, the other not, were introduced into the foregut of D. reticulatum. The subsequent movement and redistribution of the metal within the slug tissues was studied by labelling the chelates with the radioactive isotope ⁵⁵Fe. In slugs which survived treatment approximately half of the ⁵⁵Fe was voided in faeces. The iron retained became unevenly distributed, the highest concentration occurring in the digestive gland, irrespective of the chelate used. At high doses, slugs treated with tris(1-oxo-1,2-diazabutan-2-oxido)Fe(III) were fatally poisoned while those treated with the homologue, tris(1-oxo-1,2-diazaoctan-2-oxido)Fe(III) were not. Slugs killed by the toxic chelate consistently contained proportionally less iron in the digestive gland and proportionally more in the body wall and reproductive system. Dosing slugs already killed by carbon dioxide asphyxiation gave a similar pattern, suggesting that the greater mobility of the iron from the toxic chelate was not a function of the slugs' metabolism.     

Effects of short-term trimethoprim-sulfamethoxazole administration on thyroid function in dogs

OBJECTIVE: To determine how rapidly trimethoprim-sulfamethoxazole affects serum total thyroxine (T4) and thyroid-stimulating hormone (TSH) concentrations in euthyroid dogs and how quickly hormone concentrations return to reference values following discontinuation of administration. DESIGN: Prospective study. ANIMALS: 7 healthy euthyroid dogs. PROCEDURE: Dogs were given trimethoprim-sulfamethoxazole (26.5 to 31.3 mg/kg [12 to 14.2 mg/lb], PO, q 12 h) for a maximum of 6 weeks. A CBC and Schirmer tear test were performed and serum total T4 and TSH concentrations were measured weekly. Administration of trimethoprim-sulfamethoxazole was discontinued if total T4 concentration was less than the lower reference limit and TSH concentration was greater than the upper reference limit or if persistent neutropenia developed. RESULTS: Six dogs had total T4 concentrations less than the lower reference limit within 3 weeks; T4 concentration was decreased after 1 week in 3 of these 6 dogs. In these 6 dogs, TSH concentration was greater than the upper reference limit within 4 weeks. In 1 dog, T4 and TSH concentrations were not affected, despite administration of trimethoprim-sulfamethoxazole for 6 weeks. Neutropenia developed in 4 dogs. In 1 dog, the neutropenia resolved while trimethoprim-sulfamethoxazole was still being administered. In the other 3, neutrophil counts returned to reference values 1 week after drug administration was discontinued. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of trimethoprim-sulfamethoxazole at a dosage of 26.5 to 31.3 mg/kg, PO, every 12 hours can substantially alter serum total T4 and TSH concentrations and neutrophil counts in dogs within as short a time as a few weeks.     

Effects on functions of ovine blood mononuclear cells for each of several fatty acids at concentrations found in plasma of healthy and ketotic ewes

OBJECTIVE: To assess effects on functions of peripheral blood mononuclear cells (PBMC) obtained from ewes for each of several fatty acids represented in ovine plasma at concentrations mimicking those of ketotic or healthy ewes. SAMPLE POPULATION: Blood samples obtained from 6 Sardinian ewes. PROCEDURE: The PBMC were cultured in media that contained oleic (OA), palmitic (PA), stearic (SA), linoleic (LA), or palmitoleic (POA) acid at concentrations similar to those of ketotic or healthy ewes. Synthesis of DNA was stimulated by use of concanavalin A or pokeweed mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: High concentrations (900, 450, and 225 micromol/L) of OA significantly inhibited DNA synthesis and IgM secretion of PBMC. Conversely, low concentrations (56 or 28 micromol/L) of OA significantly enhanced DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, or 75 micromol/L) significantly inhibited DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, 75, or 38 micromol/L) also significantly inhibited IgM secretion of PBMC. None of the concentrations of LA and POA affected PBMC functions. CONCLUSION AND CLINICAL RELEVANCE: Impaired immunoresponsiveness of ketotic ewes is likely associated with an increase of plasma concentrations of OA, PA, or SA and not with that of LA or POA. At physiologic concentrations, single fatty acids are likely to participate in modulation of immunoresponsiveness by exerting suppressive or stimulatory effects on immune cells.     

Pluripotent stem cells--model of embryonic development,tool for gene targeting,and basis of cell therapy

Embryonic stem (ES) cells are pluripotent cell lines with the capacity of self-renewal and a broad differentiation plasticity. They are derived from pre-implantation embryos and can be propagated as a homogeneous, uncommitted cell population for an almost unlimited period of time without losing their pluripotency and their stable karyotype. Murine ES cells are able to reintegrate fully into embryogenesis when returned into an early embryo, even after extensive genetic manipulation. In the resulting chimeric offspring produced by blastocyst injection or morula aggregation, ES cell descendants are represented among all cell types, including functional gametes. Therefore, mouse ES cells represent an important tool for genetic engineering, in particular via homologous recombination, to introduce gene knock-outs and other precise genomic modifications into the mouse germ line. Because of these properties ES cell technology is of high interest for other model organisms and for livestock species like cattle and pigs. However, in spite of tremendous research activities, no proven ES cells colonizing the germ line have yet been established for vertebrate species other than the mouse (Evans and Kaufman, 1981; Martin, 1981) and chicken (Pain et al., 1996). The in vitro differentiation capacity of ES cells provides unique opportunities for experimental analysis of gene regulation and function during cell commitment and differentiation in early embryogenesis. Recently, pluripotent stem cells were established from human embryos (Thomson et al., 1998) and early fetuses (Shamblott et al., 1998), opening new scenarios both for research in human developmental biology and for medical applications, i.e. cell replacement strategies. At about the same time, research activities focused on characteristics and differentiation potential of somatic stem cells, unravelling an unexpected plasticity of these cell types. Somatic stem cells are found in differentiated tissues and can renew themselves in addition to generating the specialized cell types of the tissue from which they originate. Additional to discoveries of somatic stem cells in tissues that were previously not thought to contain these kinds of cells, they also appear to be capable of developing into cell types of other tissues, but have a reduced differentiation potential as compared to embryo-derived stem cells. Therefore, somatic stem cells are referred to as multipotent rather than pluripotent. This review summarizes characteristics of pluripotent stem cells in the mouse and in selected livestock species, explains their use for genetic engineering and basic research on embryonic development, and evaluates their potential for cell therapy as compared to somatic stem cells.     

Hyperthermia and Delayed-Onset Myopathy after Recovery from Anesthesia in a Horse

A 13-year old Thoroughbred cross gelding (528 kg) underwent lameness investigation. Because of his temperament, general anesthesia was required to facilitate ultrasound of the left fore fetlock and intra-articular medication of three joints. Anesthesia was induced with ketamine/diazepam after acepromazine/detomidine premedication. Anesthesia was maintained for 40 minutes with a guaifenesin/detomidine/ketamine intravenous infusion. Recovery from anesthesia was initially uneventful, although of a moderate duration (70 minutes). Once standing, the horse proceeded to box walk in an agitated state and became recumbent on two occasions. The horse was manually restrained, at which time its rectal temperature was 41.8°C. Cooling measures were employed (fans, ice-water enemas, wet rugs, intravenous fluid therapy (IVFT), and topical application of surgical spirit) until rectal temperature reached 38.7°C. IVFT was continued for a further 16 hours. Four days after recovery from anesthesia, bilateral triceps, deltoideus, trapezius, and rhomboideus muscle swelling was observed. Blood creatinine kinase was elevated (24,898 IU/L). Treatment for postanesthetic myopathy was initiated (hot packing of the muscle groups, topical dimethylsulfoxide [DMSO] cream application, and oral phenylbutazone). Myoglobinuria was not observed at any time. Muscle swelling decreased over the following 3 days. The horse was discharged on day 11 and has since returned to work.     

Challenges of thermal nociceptive threshold testing in the donkey

ObjectiveTo evaluate a thermal nociceptive threshold (TNT) testing device in the donkey, and the influence of potential confounding factors on TNTs.AnimalsTwo groups (Group 1 and Group 2) of eight castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsTNTs were measured by heating a thermal probe on skin until an end-point behaviour (threshold temperature) or a cut-out temperature (51 °C) was reached. The withers and the dorsal aspect of the distal limb were used as sites for TNT testing. The effects on TNT of different confounding factors: the limb tested; rate of heating; and ambient temperature were evaluated. Data were analyzed using general linear models, and Mann-Whitney tests, p < 0.05 was considered significant.ResultsEnd-point behaviours (skin twitch or donkey looking at test device) when the thermal probe heated the withers were observed in approximately half of tests. TNT was (mean ± SD) 46.8 ± 2.85 °C. Subsequently the limb was evaluated as the test site in Group 1 followed by Group 2 donkeys; end-point behaviour being a foot-lift. In Group 1, 72% of tests ended in an end-point behaviour but the response rate was lower in Group 2 (20%), although TNTs were similar [(47.6 ± 3.3) and (47.3 ± 3.0) °C respectively] for responding animals. Rate of heating, ambient temperature and laterality (right or left) did not affect thresholds, but mean TNT was significantly higher in the forelimb (48.5 ± 2.8 °C) than the hind limb (47.4 ± 2.8 °C) (p = 0.012).ConclusionsWhen a thermal probe cut-out temperature of 51 °C was used in TNT testing in the donkey a high proportion of tests did not produce an identifiable end point behaviour. Higher cut-out temperatures damaged the skin. Under these conditions, thermal nociceptive threshold testing appears not be an appropriate analgesiometry technique in the donkey.Clinical relevanceTNT testing under these conditions is not suitable form of analgesiometry for donkeys.